Minimum inhibiting concentration

Antimicrob Agents Chemother, 2002 Jun, 46(6), 1985 - 8
CTX-M-14, a plasmid-mediated CTX-M type extended-spectrum beta-lactamase isolated from Escherichia coli; Ma L et al.; Four Escherichia coli isolates harboring CTX-M-14, with a single Ala231-->Val substitution compared to CTX-M-9, had three different ribotypes . Cefotaxime resistance was plasmid encoded and conjugatively transferable . Three isolates had the same plasmid restriction enzyme digestion profile, suggesting clonal spread of a resistant plasmid . A high k(cat)/K(m) value for cefotaxime (20.3 microM(-1) s(-1)) but low values for ceftazidime and aztreonam (< 0.02 microM(-1) s(-1)) were observed in hydrolysis assays, indicating resistance to cefotaxime (MIC > or = 64 microg/ml) but susceptibility to ceftazidime (MIC < or = 2 microg/ml).

Antimicrob Agents Chemother, 2002 Jun, 46(6), 1960 - 5
Method for measuring postantifungal effect in Aspergillus species; Vitale RG et al.; An in vitro method for determination of postantifungal effect (PAFE) in molds was developed by using three isolates each of Aspergillus fumigatus, A . flavus, A . terreus, A . nidulans, and A . ustus . MICs of amphotericin B and itraconazole were determined by using National Committee for Clinical Laboratory Standards guidelines (M38-P) . The inoculum was prepared in RPMI 1640 broth buffered with MOPS (morpholinepropanesulfonic acid) at pH 7.0, and conidia were exposed to amphotericin B and itraconazole at concentrations of 4, 1, and 0.25 times the MIC, each for 4, 2, and 1 h at 37 degrees C . The same procedure was followed for controls with drug-free medium . Following exposure, the conidia were washed three times in saline and the numbers of CFU per milliliter were determined . Exposed and control conidia were then inoculated into microtitration plates and incubated at 37 degrees C for 48 h in a spectrophotometer reader . The optical density (OD) was measured automatically at 10-min intervals, resulting in growth curves . PAFE was quantified by comparing three arbitrary points in the control growth curve, the first increase of OD and the points when 20 and 50% of the maximal growth were reached, with the growth curve of drug-exposed conidia . Amphotericin B induced PAFE in A . fumigatus at four times the MIC after 2 and 4 h of exposure ranging from 1.83 to 6.00 h and 9.33 to 10.80 h, respectively . Significantly shorter PAFEs or lack of PAFE was observed for A . terreus, A . ustus, and A . nidulans . Itraconazole did not induce measurable PAFE in the Aspergillus isolates at any concentration or exposure time tested . Further studies are warranted to investigate the implications of PAFE in relation to clinical efficacy and dosing frequency.

J Med Microbiol, 2002 Jun, 51(6), 479 - 83
Voriconazole and fluconazole susceptibility of Candida isolates; Pelletier R et al.; An adapted NCCLS M27-A method was used to evaluate the activity of voriconazole (VRC) and fluconazole (FLC) against 295 Candida isolates collected from 189 patients (including isolates from deep sites) . Isolates included 186 C . albicans, 54 C . glabrata, 27 C . tropicalis, 14 C . parapsilosis, 6 C . krusei, 6 C . lusitaniae, 1 C . lypolytica and 1 C . sake . Forty-two isolates had reduced susceptibility to FLC (MIC >8 mg/L); 83.3% of these had VRC MICs < or =2 mg/L (9 of 11 C . albicans, 18 of 19 C glabrata, 6 of 6 C . krusei, 2 of 2 C . lusitaniae and 0 of 4 C . tropicalis), including 60% of isolates collected from deep-seated infections . These results suggested that in the era of azole resistance, VRC has a promising antifungal activity for serious infections with Candida spp., including most species with low susceptibility to FLC and uncommonly isolated species.

Yao Xue Xue Bao, 1998 Sep, 33(9), 650 - 4
{Effect of 8-(N,N'-diethylamino)-n-octyl-3,4,5-trimethoxybenzoate on {Ca2+}i and the release of NO in cultured endothelial cells of the calf middle cerebral artery}; Wang B et al.; The effect of 8-(N,N'-diethylamino)-n-octyl-3,4,5-trimethoxybenzoate(TMB-8) on {Ca2+}i and the release of NO in cultured endothelial cells of the calf middle cerebral artery was studied by a system of measurement of AR-CM-MIC, using Fura-2/AM as a fluorescent indicator and the oxy-haemoglobin method . In the presence of extracellular Ca2+ 1.3 mmol.L-1, the resting {Ca2+}i and the extinction difference(delta E) of between 401 and 410 nm were not changed by TMB-8 12.5 and 25 mumol.L-1, but were increased by TMB-8 50 and 100 mumol.L-1 . The rise of delta E induced by TMB-8 50 and 100 mumol.L-1 was blocked completely by L-NAME . The elevation of {Ca2+}i was related to the release of NO . The result suggested that TMB-8 increased resting {Ca2+}i of endothelial cells of the calf middle cerebral artery and induced NO release.

Ann N Y Acad Sci, 2002 Apr, 958, 341 - 4
MHC class I chain-related gene a alleles distinguish malnutrition-modulated diabetes, insulin-dependent diabetes, and non-insulin- dependent diabetes mellitus patients from eastern India; Sanjeevi CB et al.; Insulin-dependent diabetes mellitus (IDDM) is a polygenic disorder with an autoimmune basis for disease development . In addition to HLA, a second susceptibility locus for IDDM has been identified to lie in the major histocompatibility class III region . MIC-A is located in the MHC class III region and is expressed by monocytes, keratinocytes, and endothelial cells . Sequence determination of the MIC-A gene identifies trinucleotide repeat (GCT) microsatellite polymorphism in exon 5 . Five alleles with 4, 5, 6, and 9 repetitions of GCT or 5 repetitions of GCT with 1 additional nucleotide insertion (GGCT) are identified . The alleles are A4, A5, A5.1, A6, and A9 . The aim of our study was to find the association of MIC-A alleles with IDDM, malnutrition-modulated diabetes mellitus (MMDM), and non-insulin-dependent diabetes mellitus (NIDDM) patients . IDDM (n = 52), MMDM (n = 41), NIDDM (n = 212), and healthy controls (n = 73) from Cuttack, in eastern India, were studied . Of the 212 NIDDM patients analyzed, 96 of them were found to be positive for either GAD65 or IA-2 antibodies . Autoantibodies to GAD65 and IA-2 were measured by radioligand binding assay using (35)S-labeled recombinant human GAD65 and IA-2 in an in vitro transcription/translation system . Autoantibody-positive NIDDM patients (n = 96) and adult healthy controls for NIDDM (n = 113) were also compared . These autoantibody-positive NIDDM patients are considered as slow-onset IDDM or latent autoimmune diabetes in adults (LADA) patients . The samples were analyzed for MIC-A by PCR amplification, and fragment sizes were determined in an ABI prism DNA sequencer . The results of the MIC-A typing are: allele 9 of MIC-A is positively associated (OR 3.62; P < 0.001), and allele 4 is negatively associated (OR 0.31; P < 0.05) with MMDM patients compared to controls . Allele 5 is positively associated with IDDM (OR 2.64; P < 0.05) when compared to controls . Allele 5.1 is positively associated in the autoantibody-positive NIDDM patients compared to adult controls . Our findings of a significant increase of allele A9 in MMDM patients compared to healthy controls suggest that MMDM is immunogenetically different from IDDM in eastern India . MIC-A is important in the pathogenesis of MMDM patients from Cuttack . MIC-A alleles distinguish acute-onset IDDM from slow-onset IDDM, indicating that this molecule may be important for delaying the onset of IDDM with the result that these patients are diagnosed clinically as NIDDM.

Ann N Y Acad Sci, 2002 Apr, 958, 321 - 4
Contribution of MIC-A polymorphism to type 1 diabetes mellitus in Basques; Bilbao JR et al.; The maximum genetic susceptibility to type 1 diabetes (T1DM) in Basques is conferred by extended HLA haplotype F1C30-DR3-DQ2-DPB1*0202 . Due to the strong linkage disequilibrium within the haplotype, it is difficult to determine which individual allele shows the strongest association with T1DM . Recent studies of the MIC-A gene have shown an HLA-independent association of allele A5 with T1DM and A5.1 with Addison's disease . In order to test for association of the MIC-A exon 5 polymorphism with T1DM and to further characterize risk and protection haplotypes in Basques, we typed 70 Basque families with T1DM for MIC-A exon 5 polymorphism using fluorescent PCR and electrophoresis on an ABI sequencing machine . When analyzed individually, allele A4 was associated with disease {OR = 2.93 (1.58-5.5)}, while the presence of A9 conferred protection from T1DM {OR = 0.27 (0.08-0.74)} . In the context of HLA haplotypes, allele A4 was found to be associated to the F1C30-DR3-DQ2-DPB*0202 risk haplotype, both for T1DM and AFBAC alleles (P(c) = 0.0003) . Allele A5.1 was strongly associated with protective haplotype SC31-DRB1*1501-DQB1*0602, present only in AFBAC alleles, but also with risk haplotype SC01-DR3-DQ2 . In conclusion, polymorphisms in exon 5 of the MIC-A gene are associated with genetic susceptibility/protection to T1DM, but in the context of susceptibility HLA haplotypes . Nevertheless, the protective effect of A9 allele seems independent from HLA, since it does not appear to be associated with any particular extended haplotype.

Ann N Y Acad Sci, 2002 Apr, 958, 309 - 11
MHC class I chain-related gene alleles 5 and 5.1 are transmitted more frequently to type 1 diabetes offspring in HBDI families; Zake LN et al.; Type 1 diabetes mellitus (T1DM) is an autoimmune disease characterized by autoimmune destruction of pancreatic beta cells . Genetic and environmental factors contribute in this disease . There is evidence that MHC class I chain-related gene (MIC-A) plays a role in the susceptibility to this and other autoimmune diseases . There are five alleles of the MIC-A gene, which consist of different repetitions of GCT . In particular, MIC-A alleles 5 and 5.1 (the former with five repetitions of GCT, the latter with five repetitions and one additional insertion of nucleotide G) have been found to be associated with susceptibility to and age at onset of T1DM . The aim of our study was to analyze the transmission of these MIC-A alleles to T1DM-affected offsprings in HBDI families . These are multiplex families with affected offsprings and unaffected parents . DNA samples were amplified for MIC-A using fluorescence-labeled primers and analyzed on an ABI prism DNA sequencer . The transmission of alleles was then analyzed using pedigrees of families also obtained from HBDI . We analyzed 78 families and found that MIC-A alleles 5 and 5.1 are present and transmitted more frequently than expected . Heterozygotic parents for MIC-A alleles 5 and 5.1 were excluded from the study . Our results suggest that MIC-A alleles 5 and 5.1 are associated with susceptibility to T1DM in family studies.

Ann N Y Acad Sci, 2002 Apr, 958, 144 - 7
Immunogenetic studies on malnutrition-modulated diabetes mellitus; Sanjeevi CB et al.; Genetic studies of malnutrition-related diabetes are few . We have analyzed the HLA class II gene polymorphism in malnutrition-modulated diabetes mellitus (MMDM), which was previously referred to as protein-deficient diabetes mellitus (PDDM) in the 1985 WHO classification . Insulin-dependent diabetes mellitus (IDDM) is a polygenic disorder with an autoimmune basis for disease development . In addition to HLA, a second susceptibility locus for IDDM has been identified to lie in the major histocompatibility class III region . Both IDDM and MMDM in eastern Indians are associated with DR3-DQ2 but not DR4-DQ8 . The presence of autoantibodies to IDDM autoantigens in clinical MMDM either identifies the slow-onset form of IDDM or suggests autoimmunity different from that in IDDM . Our study demonstrates that the presence of GAD65 antibody and DR3-DQ2 positivity in MMDM patients identifies the underlying autoimmune mechanism in the etiology in eastern India . In autoantibody-negative MMDM patients an association with DR7-DQ2 is identified . The date obtained also indicate the possibility that MMDM can coexist with IDDM in these patients and that malnutrition could be one of the reasons for the slower onset in IDDM-prone individuals . The association of DR7-DQ2 suggests that there is a different immunogenetic background to MMDM than to IDDM . MICA is located in the MHC class I region and is expressed by monocytes, keratinocytes, and endothelial cells . Sequence determination of MICA gene identifies trinucleotide repeat (GCT) microsatellite polymorphism in exon 5 . Five alleles with 4, 5, 6, and 9 repetitions of GCT or 5 repetitions of GCT with 1 additional nucleotide insertion (GGCT) are identified . The alleles are A4, A5, A5.1, A6, and A9 . We studied the association of MICA alleles with IDDM (n = 52) and MMDM (n = 41) patients and healthy controls (n = 73) from Cuttack, eastern India . MICA was typed by PCR amplification, and fragment sizes were determined in an ABI prism DNA sequencer . Allele 9 of MICA is positively and allele 4 negatively associated with MMDM patients compared to controls . Allele 5 is positively associated with IDDM (OR 2.64, P < 0.05) when compared to controls . Our findings suggest that MMDM is immunogenetically different from IDDM in eastern India and that MIC-A is important in the pathogenesis of MMDM patients from Cuttack in eastern India.

New Microbiol, 2002 Apr, 25(2), 123 - 30
Genotypic characterization of clarithromycin-resistant Helicobacter pylori strains; Piana A et al.; Helicobacterpylori (Hp) resistance to clarithromycin, one of the antibiotics most used to eradicate infection, is connected with the presence of a point mutation on the level of adenine at position 2143 or 2144 of 23S rRNA . AIM: The aim of the study is to evaluate of the presence of these mutation vs control clarithromycin resistant Hp strains present in North Sardinia; to verify the real association between the type of mutation and the resistance-level; to use easier molecular biology methods to quickly locate the resistance-associated mutations beginning with the bioptic material . The clarithromycin susceptibility of Hp isolates was tested by the E-test method (antibiotic assay) . Genomic DNA of Hp strains was amplified using specific primers for the domain V . of ribosomic 23S rRNA and sequenced after the reaction with a primer within the fragment 23S . At the same time PCR-RFLP reliability was examined underlining the presence of these mutations with BsaI, BbsI, MboII restriction enzymes . Two mutations in 2143 (A- - G) and 2144 (A- - G) were found by domain V sequencing . The strains with mutation 2143 are characterized by a greater resistance level (MIC>64 g/ml) than those with mutation 2144 (MIC <64 g/ml) . Restriction endonucleases BbsI and MboII recognise the site containing the mutation 2143 (A- - G), while BsaI recognise the mutation 2144 (A- - G) . These methods might enable us to identify the presence of Hp directly from bioptic material and possible clarithromycin resistance and plan a suitable therapeutic strategy and consequently a better control of the infection.

Anticancer Res, 2002 Mar-Apr, 22(2A), 959 - 67
Interaction of protonated anticancer thiazines with water-insoluble phospholipids and antineoplastic agents; Flores VC et al.; A series of neuroleptic protonated phenothiazine derivatives (promethazine, promazine, triflupromazine, methotrimeprazine, propiomazine, trifluoperazine and fluphenazine), some with known anticancer properties, were complexed with water-insoluble antineoplastice agents such as 5-fluorouracil (5FU), methotrexate (MTX) and sulindac, as well as with components of biomembrane and synthetic phospholipids as possible models of cancer and microbial cells . In all cases water-soluble micellar inclusion adducts were formed exhibiting electron charge transfer complex behaviour, with the appearance of thazine free radicals . The thiazines sequestered the drugs and phospholipids in well-defined molar ratios (MR) parabolically-dependent on the dipole moments (mu) of the protonated phenothiazine derivatives . pH comparisons showed that the inclusion adducts followed a model in which the compounds were enveloped in the lipophilic interior of the thiazine aggregates, while the side-chains of the latter faced the aqueous environment . In the model experiment, interaction of the thiazines and the 5FU adducts with E . coli F' lac was additive to marginally synergistic, confirmed by the checkerboard technique . The parabolic dependence of the molar minimum inhibitory concentration of the thiazines and thiazine/5FU adducts on the thiazine dipole moments suggests that their primary loci of interaction are the cell wall or membrane phospholipid components . Thiazines, especially those with dipole moments centered on about 6 D, near which the lowest MR occurs, can act as effective carriers for insoluble or sparsely soluble drugs . Any new thiazine drug for use alone or as a carrier in anticancer therapy should be designed with this criterion in mind.

Int J Antimicrob Agents, 2002 May, 19(5), 389 - 96
Change of pneumococcal resistance to antibiotics in adults between 1995 and 1997: a study in eight French counties; Roussel-Delvallez M et al.; The main object of this study was to describe the evolution of antibiotic resistance in pneumococci from adults, in eight French counties of France between 1995 and 1997 . Despite the high and increasing prevalence (23-35%) of pneumococci with diminished susceptibility to penicillin G (PSDP), resistance to amoxycillin (0.8-0.5%) and to cefotaxime (0-0.3%) was rare in both 1995 and 1997 respectively . The percentage of pneumococci resistant to penicillin G (PRP, minimum inhibitory concentration >1 mg/l) remained stable between the two periods . PSDP showed increased resistance to macrolides (30-41%), to cotrimoxazole (28-34%) and to tetracycline (19-25%) . These figures are lower than those obtained over the same periods and the same regions in children . The distribution of PSDP serotypes isolated in adults was the same as that seen in children: by descending order serotypes 23, 14, 9 and 6 . This study by the Regional Pneumococcal Observatories confirms the high prevalence and the main characteristics of antibiotic resistance among pneumococci with variations in levels of resistance with the age of patients, with the site of sampling and from one Observatory to another.

J Ethnopharmacol, 2002 May, 80(2-3), 193 - 7
Studies on antimycotic properties of Datura metel; Rajesh et al.; The hexane, chloroform, acetone and methanolic fractions of Datura metel L . were investigated for antifungal properties using pathogenic species of Aspergillus . The chloroform fraction was found to be endowed with antifungal activity . The minimum inhibitory concentration (MIC) of chloroform fraction of D . metel L . was 625.0 microg ml(-1) against all the three species of Aspergillus, i.e . A . fumigatus, A . flavus and A . niger, by microbroth dilution and percent spore germination inhibition assays . The MIC by disc diffusion assay was observed to be 12.5 microg disc(-1) . The chloroform fraction of D . metel, when investigated for potency, turned to be 9.2 times less active than amphotericin B . However, it was important to note that the cytotoxicity of chloroform fraction in vitro was 117.8 times less than amphotericin B.

Pediatr Infect Dis J, 2002 Mar, 21(3), 234 - 40
Comparison of two gentamicin dosing schedules in very low birth weight infants; Rastogi A et al.; BACKGROUND: Several dosing schedules for gentamicin have been recommended for very low birth weight infants during the early neonatal period . We conducted a prospective, randomized, controlled trial to compare efficacy and pharmacokinetics of two dosing schedules in preterm neonates . METHODS: Fifty-eight very low birth weight infants (600 to 1500 g), prescribed gentamicin for treatment of suspected sepsis during the first week after birth, were randomized to receive either the new dosing schedule {every 48 h (q48h)} or the existing dosing schedule {every 24 h (q24h)} . Infants in the "q48h" group received gentamicin at 5.0 or 4.5 mg/kg/dose q48h depending on weight group and infants in the "q24h" group received 2.5 or 3.0 mg/kg/dose q24h . Peak and trough serum gentamicin concentrations were monitored . RESULTS: Peak serum gentamicin concentrations after the first dose were significantly higher in the q48h infants than in q24h infants (8.19 +/- 1.3 vs . 6.04 +/- 2.2, P = 0.00001) . Ninety percent of all peak serum gentamicin concentrations in the q48h group were in a higher therapeutic range of 6 to 12 microg/ml as compared with 55% of q24h (P = 0.0005) . None of the q48h infants had subtherapeutic serum gentamicin concentrations immediately after administration of the first dose as compared with 36% of q24h infants (P < 0.005) . Eighteen percent of q24h infants continued to have peak serum gentamicin concentrations in subtherapeutic range even after the third dose at 48 h . Trough serum gentamicin concentrations were significantly lower in q48h infants than in q24h infants . However, 9 of 30 (30%) q48h infants had trough serum gentamicin concentrations of < or = 0.5 microg/ml before the dose at 48 h and 4 of the 9 had serum gentamicin concentrations of <1 microg/ml at 24 h after the first dose . CONCLUSIONS: The q48h dosing schedule of gentamicin given to very low birth weight infants during the first week after birth achieved therapeutic serum gentamicin concentrations and potentially higher peak to MIC ratios for microorganisms in all infants . However, nearly one-third of the infants had extremely low serum gentamicin concentrations before the next dose . A dosing interval of 36 h might be optimal for bactericidal activity and avoid bacterial growth during prolonged periods of extremely low serum gentamicin concentrations; this dosing interval warrants study.

J Antimicrob Chemother, 2002 May, 49(5), 757 - 61
Serial passage of Chlamydia spp . in sub-inhibitory fluoroquinolone concentrations; Morrissey I et al.; We investigated the in vitro development of fluoroquinolone resistance in Chlamydia trachomatis and Chlamydia (Chlamydophila) pneumoniae grown in McCoy cell monolayers in supplemented Eagle's minimum essential medium . With C . trachomatis, initial passages at sub-inhibitory fluoroquinolone concentrations did not affect fluoroquinolone susceptibility . However, after an initial lag of 10-24 passages (depending upon the fluoroquinolone used), fluoroquinolone resistance developed rapidly . The final fluoroquinolone MIC after a total of 30 passages was >256 times the MIC of the original wild-type strain with ofloxacin or ciprofloxacin passage . Analysis of the quinolone-resistance determining regions of two quinolone-resistant C . trachomatis mutants obtained after 30 passages showed that both isolates had a single serine to isoleucine substitution at amino acid position 83 in GyrA . In stark contrast, with C . pneumoniae no reduced fluoroquinolone susceptibility could be sustained, even after 30 passages with moxifloxacin or ofloxacin . With sparfloxacin passage, some indication of resistance was observed but no viable organisms could be isolated for further investigation . It is possible that fluoroquinolone-resistant C . pneumoniae are less able to survive than wild type, which may explain why resistance does not develop readily.

J Antibiot (Tokyo), 2002 Feb, 55(2), 128 - 33
Kosinostatin, a quinocycline antibiotic with antitumor activity from Micromonospora sp . TP-A0468; Furumai T et al.; Kosinostatin, a quinocycline antibiotic was isolated from the culture broth of an actinomycete strain TP-A0468 along with isoquinocycline B . The producing strain was isolated from the seawater sample collected in Toyama Bay and identified as Micromonospora sp . based on the taxonomic study . Kosinostatin was obtained from the culture fluid by solvent extraction and ODS column chromatography . Kosinostatin inhibited the growth of Gram-positive bacteria strongly (MIC=0.039 microg/ml) and Gram-negative bacteria and yeasts moderately (MIC= 1.56 approximately 12.5 microg/ml) . It showed cytotoxicity against various cancer cell lines with the IC50 of 0.02 approximately 0.6 microm and inhibited human DNA topoisomerase Ila with the IC50 of 3 approximately 10 microM.

Epidemiol Infect, 2002 Apr, 128(2), 337 - 42
Pyrazinamide resistance associated with pncA gene mutation in Mycobacterium tuberculosis in Japan; Endoh T et al.; Thirty Japanese clinical isolates of Mycobacterium tuberculosis were analysed by pyrazinamide susceptibility testing and pyrazinamidase assay, as well as polymerase chain reaction for single-strand conformational polymorphism and direct sequencing of the gene encoding pyrazinamidase (pncA) . All sensitive isolates showed pyrazinamidase activity and a wild-type pncA gene, but three resistant isolates had pncA gene mutations and lacked pyrazinamidase activity . The latter isolates showed a minimum inhibitory concentration of at least 100 mg/l by the 7H10 agar proportion method and 400 mg/l by the 7H9 liquid medium method . Isolate 28 showed T-to-C change at position 11, leading to Leu4 --> Ser substitution; isolate 29 had an 8-bp deletion from position 382; and isolate 30 had A-to-C change at position 29, leading to Gln10 --> Pro substitution . The deletion has not been described previously . This is the first demonstration of pncA gene mutations in PZA-resistant M . tuberculosis strains isolated from Japanese patients.

J Appl Microbiol, 2002, 92 Suppl, 78S - 84S
Antibiotic exposure as a risk factor for emergence of resistance: the influence of concentration; Gould IM et al.; Evolution of antibiotic resistance (AR) is increasingly perceived as a major clinical problem . The use of bactericidal antibiotics may protect against this, to some extent, by eradication of the pathogen, but the borders between cidal and inhibitory activity in the patient are often blurred . In addition, there are clinical reasons why eradication of the pathogen may not always be desirable . Antibiotic dosing schedules are currently driven by the perception that T >MIC and AUIC are the main predictors of outcome for time-dependent and concentration-dependent antibiotics, respectively . In the context of protecting against development of resistance in the pathogen however, peak antibiotic concentration and the concept of mutant prevention concentrations may be more important . The role of post-antibiotic and sub-MIC effects is more conjectural . Considerations of mechanisms of resistance and their relationship to antibiotic dosing schedules will also be highlighted . Lastly, the relevance of all this to the development of resistance in the normal bacterial flora will be discussed.

Enferm Infecc Microbiol Clin, 2002 Apr, 20(4), 157 - 60
{Evolution of the sensitivity of 235 strains of Helicobacter pylori from 1995 to 1998 and impact of antibiotic treatment}; Cuchi Burgos E et al.; BACKGROUND: The aim of this study was to investigate the sensitivity of Helicobacter pylori to the antibiotics used in its eradication over a period of four years and to determine the influence of previous treatment on sensitivity . MATERIAL AND METHODS: During the period from 1995 to 1998 we determined the sensitivity of 235 consecutive Helicobacter pylori isolates to amoxicillin, metronidazole, clarythromycin and tetracycline by means of E-test methodology . The MIC values found were related with the prior use of eradicating treatment . RESULTS: The percentage of resistant strains were as follows: 23.5% to metronidazole, 12.9% to clarythromycin and 0.7% to tetracycline; none of the strains was resistant to amoxicillin . There were no significant changes in percentage of resistance to the drugs studied over the 4-year period . Resistance to metronidazole and clarythromycin was significantly higher (p 5 0.03 and p < 0.001 respectively) in strains isolated from patients who had received previous treatment . CONCLUSIONS: Monitorization of H . pylori sensitivity to the drugs used in its eradication is particularly important in patients who have undergone prior treatment.

J Asthma, 2002 Apr, 39(2), 143 - 50
Forced expiratory time and bronchial hyperresponsiveness to methacholine; Goldstein MF et al.; Pulmonary junction tests (PFTs) are normally performed prior to methacholine inhalation challenges (MICs) . In contrast to normal baseline spirometry (FEV1, FEF25%-27%, FVC), we have observed patients with positive MICs having shortened forced expiratory times (FET100%) in the baseline pre-MIC PFT . We prospectively evaluated the correlation of abnormalities in baseline pre-MIC FET100% in patients who have positive vs . negative MICs . Prospective analysis of baseline pre- MIC FET100%, and MIC results in suspected asthmatics with normal lung exams, spirometry and chest x-rays . Using a PC20 FEV1 of < or =8mg/ml methacholine there were 115 positive and 69 negative MICs . The mean (+/-1 SD) FET100% in the positive MIC group was 3.57+/-1.68 sec vs . 4.73+/-1.60 sec in the negative group . The difference in these means was statistically significant (p <0.0001) . There was a statistically significant difference in the incidence of FET100% <4sec in the positive (55.65%) vs . the negative (30.43%) MIC group, p<0.001 . There was also a statistically significant difference in the incidence of positive MIC in FET100% <4sec (75.29%) vs . FET100%, > or =4sec (51.52%), p< 0.001 . Our results suggest that in our highly selected, well-characterized population, FET100.% <6sec is common and FET100% <4 sec correlates with an increased likelihood of having a positive MIC.

Farmaco, 2002 Apr, 57(4), 259 - 65
New benzimidazole derivatives as antimycobacterial agents; Klimesova V et al.; A set of 2-alkylsulfanyl derivatives of 5-methylbenzimidazole was synthesized and evaluated for antimycobacterial activity . The structures of the compounds were confirmed by 1H NMR and IR data, and their purity by elemental analysis . Antimycobacterial activities against Mycobacterium tuberculosis and nontuberculous mycobacteria were expressed as the minimum inhibitory concentration . The substances exhibited significant antimycobacterial activity, in particular against both strains of Mycobacterium kansasii . The effect of the most active compound in the set, 3,5-dinitro derivative 3t, exceeded that of the standard isoniazide against M . kansasii and Mycobacterium avium.

Thorac Cardiovasc Surg, 2002 Apr, 50(2), 87 - 91
Mediastinitis and cardiac surgery--an updated risk factor analysis in 10,373 consecutive adult patients; Gummert JF et al.; BACKGROUND: Deep sternal wound infection (DSWI) remains a serious complication after cardiac surgery . New evolving techniques including the utilization of internal mammary arteries (IMA), beating heart procedures, and minimal invasive surgery (MIC) require an updated risk factor analysis to identify high risk patients in order to improve perioperative treatment . METHODS: 10,373 consecutive patients receiving cardiac surgery between May 1996 and August 1999 were evaluated: 9,303 underwent full sternotomy whereas a minimally invasive (MIC) approach using partial sternotomy or lateral thoracotomy was used in 1,070 patients . DSWI was defined as the evidence of mediastinitis seen at reoperation along with one or more of the following: positive culture of mediastinal fluid, positive blood culture or temperature higher than 38 degrees C and/or leukocytosis . RESULTS: The overall incidence of DSWI in the "full sternotomy" group was 1.44 % (134 of 9,303) . Univariate risk factor analysis showed a significant influence of IMA use, ICU / IC treatment > 5 days, postoperative ventilator time > or = 72 h, need for reexploration, diabetes, surgery time > or = 180 min, assist device implantation (including use of IABP), peripheral vascular disease and increased body mass index . Multivariate analysis identified double IMA, ICU treatment > 5 days, single IMA, diabetes, reexploration and increased body mass as significant risk factors . No mediastinitis was observed in the MIC group . CONCLUSION: As DSWI is related to sternotomy, a MIC approach should be considered for patients at high risk for DSWI . IMA takedown as a pedicled graft should be especially avoided in patients with diabetes since the risk for postoperative mediastinitis is unacceptably high in this patient group.

J Gen Physiol, 2002 May, 119(5), 487 - 507
Separation and characterization of currents through store-operated CRAC channels and Mg2+-inhibited cation (MIC) channels; Prakriya M et al.; Although store-operated calcium release-activated Ca(2+) (CRAC) channels are highly Ca(2+)-selective under physiological ionic conditions, removal of extracellular divalent cations makes them freely permeable to monovalent cations . Several past studies have concluded that under these conditions CRAC channels conduct Na(+) and Cs(+) with a unitary conductance of approximately 40 pS, and that intracellular Mg(2+) modulates their activity and selectivity . These results have important implications for understanding ion permeation through CRAC channels and for screening potential CRAC channel genes . We find that the observed 40-pS channels are not CRAC channels, but are instead Mg(2+)-inhibited cation (MIC) channels that open as Mg(2+) is washed out of the cytosol . MIC channels differ from CRAC channels in several critical respects . Store depletion does not activate MIC channels, nor does store refilling deactivate them . Unlike CRAC channels, MIC channels are not blocked by SKF 96365, are not potentiated by low doses of 2-APB, and are less sensitive to block by high doses of the drug . By applying 8-10 mM intracellular Mg(2+) to inhibit MIC channels, we examined monovalent permeation through CRAC channels in isolation . A rapid switch from 20 mM Ca(2+) to divalent-free extracellular solution evokes Na(+) current through open CRAC channels (Na(+)-I(CRAC)) that is initially eightfold larger than the preceding Ca(2+) current and declines by approximately 80% over 20 s . Unlike MIC channels, CRAC channels are largely impermeable to Cs(+) (P(Cs)/P(Na) = 0.13 vs . 1.2 for MIC) . Neither the decline in Na(+)-I(CRAC) nor its low Cs(+) permeability are affected by intracellular Mg(2+) (90 microM to 10 mM) . Single openings of monovalent CRAC channels were not detectable in whole-cell recordings, but a unitary conductance of 0.2 pS was estimated from noise analysis . This new information about the selectivity, conductance, and regulation of CRAC channels forces a revision of the biophysical fingerprint of CRAC channels, and reveals intriguing similarities and differences in permeation mechanisms of voltage-gated and store-operated Ca(2+) channels.

J Clin Microbiol, 2002 May, 40(5), 1879 - 81
Candida glabrata oropharyngeal candidiasis in patients receiving radiation treatment for head and neck cancer; Redding SW et al.; Candida glabrata colonization is common in patients receiving radiation treatment for head and neck cancer, but to our knowledge has never been described as the infecting organism with oropharyngeal candidiasis (OPC) . This study presents the first three patients described with C . glabrata OPC in this patient population . Patient 1 developed C . glabrata OPC and required fluconazole, 800 mg/day, for clinical resolution . Antifungal susceptibility testing revealed a MIC of fluconazole of >64 microg/ml . Elapsed time from initial culturing to treatment decision was 7 days . Patients 2 and 3 developed C . glabrata OPC . They were patients in a study evaluating OPC infections, and cultures were taken immediately . CHROMagar Candida plates with 0, 8, and 16 microg of fluconazole/ml were employed for these cultures . Lavender colonies, consistent with C . glabrata, grew on the 0- and 8-microg plates but not on the 16-microg plate from patient 2 and grew on all three plates from patient 3 . Based on these data, a fluconazole dose of 200 mg/day was chosen for patient 2 and a dose of 400 mg/day was chosen for patient 3, with clinical resolution in both . Elapsed time from initial culturing to treatment decision was 2 days . C . glabrata does cause OPC in head and neck radiation treatment patients, and the use of fluconazole-impregnated chromogenic agar may significantly reduce treatment decision time compared to that with conventional culturing and antifungal susceptibility testing.

J Clin Microbiol, 2002 May, 40(5), 1831 - 3
In vitro activities of terbinafine in combination with fluconazole, itraconazole, voriconazole, and posaconazole against clinical isolates of Candida glabrata with decreased susceptibility to azoles; Perea S et al.; A checkerboard microdilution method, performed according to the recommendations of the National Committee for Clinical Laboratory Standards, was used to study the in vitro interaction of terbinafine (TRB) with fluconazole (FLU), itraconazole (ITRA), voriconazole (VRC), and posaconazole (PSZ) in 24 isolates of Candida glabrata with decreased susceptibility to azoles isolated from the oral cavities of human immunodeficiency virus patients . Synergy, defined as a fractional inhibitory concentration index of < or =0.5, was observed in 17% of TRB-FLU interactions, 21% of TRB-ITRA interactions, 33% of TRB-VRC interactions, and 12% of TRB-PSZ interactions . Where synergy was not achieved, there was still a decrease in the MIC of one or both drugs when used in combination . Antagonism was not observed in any drug combination . Clinical studies are warranted to elucidate the potential utility of these combination therapies.

J Clin Microbiol, 2002 May, 40(5), 1694 - 7
Clinical evaluation of a frozen commercially prepared microdilution panel for antifungal susceptibility testing of seven antifungal agents, including the new triazoles posaconazole, ravuconazole, and voriconazole; Pfaller MA et al.; A commercially prepared frozen broth microdilution panel (Trek Diagnostic Systems, Westlake, Ohio) was compared with a reference microdilution panel for antifungal susceptibility testing of two quality control (QC) strains and 99 clinical isolates of Candida spp . The antifungal agents tested included amphotericin B, flucytosine, fluconazole, itraconazole, posaconazole, ravuconazole, and voriconazole . Microdilution testing was performed according to NCCLS recommendations . MIC endpoints were read visually after 48 h of incubation and were assessed independently for each microdilution panel . The MICs for the QC strains were within published limits for both the reference and Trek microdilution panels . Discrepancies among MIC endpoints of no more than 2 dilutions were used to calculate the percent agreement . Acceptable levels of agreement between the Trek and reference panels were observed for all antifungal agents tested against the 99 clinical isolates . The overall agreement for each antifungal agent ranged from 96% for ravuconazole to 100% for amphotericin B . The Trek microdilution panel appears to be a viable alternative to frozen microdilution panels prepared in-house.

Pathol Biol (Paris), 2002 Apr, 50(3), 178 - 83
{The Pneumococcal Observatory for the Central Region, 1 April 1999 to 31 March 2000}; Cattier B et al.; Seven hundred and ninety six strains of pneumococcus were collected in the Centre region, from 15 laboratories, between 1st April 1999 and 31st of March 2000 . Data were processed, using 4th dimension software, and concerned age, file number, consultation/hospitalisation, sample type, susceptibility to oxacillin (5 micrograms), results of the E-test for benzylpenicillin, amoxicillin, cefotaxime and results of the routine disc diffusion test . Strains with reduced susceptibility to benzylpenicillin (PRSP) were collected by the co-ordinating centre to perform MICs by the reference agar dilution test and serotyping . Out of 796 strains, 450 strains (56.7%) were categorised as PRSP and 400 of them were studied by the co-ordinating centre . Forty two percent of the samples originated from lungs, followed by 19.5% from blood samples, 15% from ear pus (85.7% PRSP) and 2.5% from CSF . Thirty nine percent of the patients were female . 36.6% were children under sixteen (70.1% PRSP) and 62.4% were adults (49.2% PRSP) . Out of 400 PRSP 106 (26.5%) were characterised as resistant and 294 (73.5%) as intermediate to benzylpenicillin . Compared to the agar dilution test, 90% of the PRSP studied by E-test had a MIC value for benzylpenicillin within +/- 1 dilution . Thirty six strains of PRSP were resistant to amoxicillin (9% of the PRSP) and 10 (2.5% of the PRSP) to cefotaxime . Serotyping was done on 375 strains . The serotypes encountered were the following: 23 (26.9%), 14 (22.1%), 19 (19.5%), 6 (12.8%), 9 (9.9%) and 15 (5.1%).

Nippon Shokakibyo Gakkai Zasshi, 2002 Apr, 99(4), 379 - 85
{Anti-Helicobacter pylori effects of Bainiku-ekisu (concentrate of Japanese apricot juice)}; Fujita K et al.; Helicobacter pylori (H.p.) bacteria are the major causes of gastro-duodenal disease, and some association with stomach cancer has been suggested . Recently, H.p . eradicating treatment has been practiced using antibiotics and proton-pump inhibitors . However, at the same time, some reports have been made on the side-effects of this treatment; allergic reactions and uneffective resistant bacteria . Under these circumstances, there is a strong need for medicines, which are less harmful to the body, can be administered repeatedly, are less expensive, and yet as effective as antibiotics in inhibiting the bacteria, and in fact, some studies have been undertaken in this regard . We placed our focus on Bainiku-ekisu (Concentrate of Japanese apricot juice) which, as a Japanese folk remedy, has been used for the treatment of gastritis and enteritis since ancient times, and studied its bacteriosterile affects . The major ingredients of Bainiku-ekisu are citric acid (32%) and malic acid (11%), while its pH represents strong acid . We measured the bacteriosterile effects of Bainiku-ekisu by culturing ten H.p . strains originating from gastro mucous membrane respectively in a Bainiku-ekisu concentration of 0.156%, 0.313%, 0.625% and 0.9%, and measured the level of MIC (minimum inhibiting concentration of development) . As a result, out of ten H.p . strains four of them presented strong bacteriosterile effects in a concentration of less than 0.156%, and six of them, in a concentration of less than 0.313% . Furthermore, in order to measure the bacteriosterile effects against H.p . in the stomach we measured the quantity of bacteria in 0 minutes, five minutes, and ten minutes after mixing ten H.p . strains suspended in physiological salt solution with a Bainiku-ekisu solution of 0.3% and 0.9% dissolved in aseptic physiological salt solution . As a result, within five minutes after mixing, every one of the ten H.p . strains was observed to present strong bacteriosterile effects . These results suggest that Bainiku-ekisu can be considered as a fool likely to prevent the development of H.p . as well as the future possibility of making clinical applications to H.p . infection.

Int J Antimicrob Agents, 2002 Apr, 19(4), 323 - 31
Breakpoints: current practice and future perspectives; Mouton JW; Much of the discussion over the past decades on the value and setting of breakpoints has been due to the fact that the breakpoint was used in two ways; as an indicator to predict the probability of clinical success and also to detect resistant (sub) populations . It is apparent that these two meanings have lead to a different approach to setting, interpretation and use of breakpoints based on clinical efficacy on the one hand and breakpoints based on detection of resistance on the other . Nevertheless, several of the current guidelines make no perceptible distinction between these two meanings . A case is therefore strongly made to recognize that there is a difference between clinical and microbiological breakpoints . The microbiological breakpoint may be used to detect organisms that do not belong to the natural bacterial population, but somehow have acquired resistance and might be useful in recognizing emergence of resistant subpopulations and may lead to subsequent measures to be taken . Alternatively, the clinical breakpoint is of principal value to the clinician in that it results in a classification of S (susceptible), I (intermediate susceptible) and R (resistant) and is used in clinical practice and correlate with a measure of clinical efficacy . Methods developed during the last few years to arrive at meaningful clinical breakpoints are discussed, such as CART analysis and Monte Carlo simulation . In discussing future developments, it is suggested that current reports containing S, I, and R be at least supplemented with the MICs measured and, using current techniques available such as Monte Carlo simulation, provide the probability of successful eradication of the micro-organism and successful treatment based on population pharmacokinetics and Minimal Inhibitory Concentration (MIC) distributions.

Hum Immunol, 2002 May, 63(5), 418 - 23
Major histocompatibility complex class I chain related (MIC) A gene, TNFa microsatellite alleles and TNFB alleles in juvenile idiopathic arthritis patients from Latvia; Nikitina Zake L et al.; In order to analyze involvement of major histocompatibility complex class I chain-related gene A (MICA) and tumor necrosis factor a (TNFa) microsatellite polymorphisms as well as TNFB gene in juvenile idiopathic arthritis (JIA), we studied 128 patients divided into groups according to clinical features {monoarthritis (n = 14), oligoarthritis (n = 58), polyarthritis (n = 50), and systemic (n = 6)}, and 114 age- and sex-matched healthy controls from Latvia . DNA samples were amplified with specific primers and used for genotyping of MICA and TNFa microsatellite . Typing for a biallelic NcoI polymerase chain reaction RFLP polymorphism located at the first intron of TNFB gene was done as follows: restriction digests generated fragments of 555bp and 185bp for TNFB*1 allele, and 740bp for TNFB*2 allele . The results were compared between cases and controls . We found significant increase of MICA allele A4 (p = 0.009; odds ratio {OR} = 2.3) and allele TNFa2 (p = 0.0001; OR = 4.4) in patients compared with controls . The frequency of allele TNFa9 was significantly decreased (p = 0.0001; OR = 0.1) in patients with JIA . No significant differences of TNFB allele frequency were found . Our data suggest that MICA and TNFa microsatellite polymorphisms may be used as markers for determination of susceptibility and protection from JIA.

Akush Ginekol (Sofiia), 1999, 38(1), 46 - 9
{Grading intraductal breast carcinoma}; Popovska S et al.; Mammographic screening has led to increased detection of intraductal carcinoma of the breast (IdC) . IdC is y heterogeneous disorder with different clinical outcomes . The study was designed to confirm consensibility of the three-tiered classification, accepted by Thomas Jefferson University Consensus Conference since 1997 . The study population comprised 30 patients with IdC including patients with T1 mic and T1a . Sixty percent of IdC were classified as II-nd grade, 27% were III-rd grade and 13% as I-st grade . 20% of IdC exhibited heterogenity of grade . The cases with cribriform and papillary architecture were 1-st or II-nd grades . The other cases with comedo and solid architecture we are II-nd or III-rd grades . The cases with invasive component (T1 mic u T1a) were 25%--I-st grade, 75%--II-nd grade and III-rd grade--87.5% . In conclusion, the grading of IdC could y prognostic factor in far long term prognosis of patients with those lesions.

Phytomedicine, 1999 Nov, 6(5), 341 - 5
Antitubercular activity of pentacyclic triterpenoids from plants of Argentina and Chile; Wachter GA et al.; Screening of plants from South America for antitubercular activity and subsequent assay-guided fractionation resulted in the isolation and characterization of several pentacyclic triterpenoids . The MIC values of 22 triterpenoids were determined using the radiorespiratory BACTEC assay and range from 8 microM to above 128 microM . The structure-activity relationships are discussed.

Antimicrob Agents Chemother, 2002 May, 46(5), 1583 - 5
In vitro activities of free and lipid formulations of amphotericin B and nystatin against clinical isolates of Coccidioides immitis at various saprobic stages; Gonzalez GM et al.; We investigated the susceptibilities of hyphal, mixed hyphal, ungerminated arthroconidial, and germinated arthroconidial populations of Coccidioides immitis to lipid formulations of amphotericin B and nystatin and their conventional preparations, utilizing the National Committee for Clinical Laboratory Standards M38-P broth macrodilution method . The differences in effects of the three different growth stages of the saprobic phase of C . immitis on the MIC/minimum lethal concentration (MLC) ratio were not statistically significant for any of the antifungal agents tested . These results suggest that either inocula could be used for in vitro susceptibility studies with C . immitis.

Antimicrob Agents Chemother, 2002 May, 46(5), 1564 - 7
Association of metronidazole resistance and natural competence in Helicobacter pylori; Yeh YC et al.; To study whether the capability of horizontal DNA transfer is associated with metronidazole resistance in Helicobacter pylori, a total of 81 clinical isolates were tested for MICs of metronidazole (MTZ) . The MIC assays were performed by using the E-test and reconfirmed by the agar dilution method . Natural competence assays were performed by transferring a chloramphenicol acetyltransferase cassette and a 23S rRNA gene from a clarithromycin-resistant strain (with an A-to-G mutation at nucleotide 2143) by using natural transformation . Of the 81 isolates, 65 (80.2%) were naturally competent while 16 were not . Among the 65 naturally competent strains, 39 (60%) were highly resistant to MTZ (MICs, >32 microg/ml) while only 2 of 16 (12.5%) noncompetent strains were highly MTZ resistant (P, <0.001) . Therefore, there is an association between natural competence and MTZ resistance.

Antimicrob Agents Chemother, 2002 May, 46(5), 1475 - 80
Intrapulmonary pharmacokinetics of linezolid; Conte JE Jr et al.; In this study, our objective was to determine the steady-state intrapulmonary concentrations and pharmacokinetic parameters of orally administered linezolid in healthy volunteers . Linezolid (600 mg every 12 h for a total of five doses) was administered orally to 25 healthy adult male subjects . Each subgroup contained five subjects, who underwent bronchoscopy and bronchoalveolar lavage (BAL) 4, 8, 12, 24, or 48 h after administration of the last dose . Blood was obtained for drug assay prior to administration of the first dose and fifth dose and at the completion of bronchoscopy and BAL . Standardized bronchoscopy was performed without systemic sedation . The volume of epithelial lining fluid (ELF) recovered was calculated by the urea dilution method, and the total number of alveolar cells (AC) was counted in a hemocytometer after cytocentrifugation . Linezolid was measured in plasma by a high-pressure liquid chromatography (HPLC) technique and in BAL specimens and AC by a combined HPLC-mass spectrometry technique . Areas under the concentration-time curves (AUCs) for linezolid in plasma, ELF, and AC were derived by noncompartmental analysis . Half-lives for linezolid in plasma, ELF, and AC were calculated from the elimination rate constants derived from a monoexponential fit of the means of the observed concentrations at each time point . Concentrations (means +/- standard deviations) in plasma, ELF, and AC, respectively, were 7.3 +/- 4.9, 64.3 +/- 33.1, and 2.2 +/- 0.6 microg/ml at the 4-h BAL time point and 7.6 +/- 1.7, 24.3 +/- 13.3, and 1.4 +/- 1.3 microg/ml at the 12-h BAL time point . Linezolid concentrations in plasma, ELF, and AC declined monoexponentially, with half-lives of 6.9, 7.0, and 5.7 h, respectively . For a MIC of 4, the 12-h plasma AUC/MIC and maximum concentration/MIC ratios were 34.6 and 3.9, respectively, and the percentage of time the drug remained above the MIC for the 12-h dosing interval was 100%; the corresponding ratios in ELF were 120 and 16.1, respectively, and the percentage of time the drug remained above the MIC was 100% . The long plasma and intrapulmonary linezolid half-lives and the percentage of time spent above the MIC of 100% of the dosing interval provide a pharmacokinetic rationale for drug administration every 12 h and indicate that linezolid is likely to be an effective agent for the treatment of pulmonary infections.

Antimicrob Agents Chemother, 2002 May, 46(5), 1455 - 61
Potato dextrose agar antifungal susceptibility testing for yeasts and molds: evaluation of phosphate effect on antifungal activity of CMT-3; Liu Y et al.; The broth macrodilution method (BMM) for antifungal susceptibility testing, approved by the National Committee for Clinical Laboratory Standards (NCCLS), was found to have deficiencies in testing of the antifungal activity of a new type of antifungal agent, a nonantibacterial chemically modified tetracycline (CMT-3) . The high content of phosphate in the medium was found to greatly increase the MICs of CMT-3 . To avoid the interference of phosphate in the test, a new method using potato dextrose agar (PDA) as a culture medium was developed . Eight strains of fungi, including five American Type Culture Collection strains and three clinical isolates, were used to determine the MICs of amphotericin B and itraconazole with both the BMM and the PDA methods . The MICs of the two antifungal agents determined with the PDA method showed 99% agreement with those determined with the BMM method within 1 log(2) dilution . Similarly, the overall reproducibility of the MICs with the PDA method was above 97% . Three other antifungal agents, fluconazole, ketoconazole, and CMT-3, were also tested in parallel against yeasts and molds with both the BMM and the PDA methods . The MICs of fluconazole and ketoconazole determined with the PDA method showed 100% agreement within 1 log(2) dilution of those obtained with the BMM method . However, the MICs of CMT-3 determined with the BMM method were as high as 128 times those determined with the PDA method . The effect of phosphate on the antifungal activity of CMT-3 was evaluated by adding Na2HPO4 to PDA in the new method . It was found that the MIC of CMT-3 against a Penicillium sp . increased from 0.5 microg/ml (control) to 2.0 microg/ml when the added phosphate was used at a concentration of 0.8 mg/ml, indicating a strong interference of Na2HPO4 with the antifungal activity of CMT-3 . Except for fluconazole, all the other antifungal agents demonstrated clear end points among the yeasts and molds tested . Nevertheless, with its high reproducibility, good agreement with NCCLS proposed MIC ranges, and lack of interference of phosphate, the PDA method shows promise as a useful assay for antifungal susceptibility testing and screening for new antifungal agents, especially for drugs that may be affected by high (supraphysiologic) phosphate concentrations.

Antimicrob Agents Chemother, 2002 May, 46(5), 1381 - 7
Nonparametric population pharmacokinetic analysis of amikacin in neonates, infants, and children; Treluyer JM et al.; The therapeutic and toxic effects of amikacin are known to depend on its concentration in plasma, but the pharmacokinetics of this drug in neonates, infants, and children and the influences of clinical and biological variables have been only partially assessed . Therapeutic drug monitoring data collected from 155 patients (49 neonates, 77 infants, and 29 children) receiving amikacin were analyzed by a nonparametric population-based approach, the nonparametric maximum-likelihood method . We assessed the effects of gestational and postnatal age, weight, Apgar score, and plasma creatinine and urea concentrations on pharmacokinetic parameters . There is no specific formulation of amikacin for neonates and infants . We therefore used an error model to account for errors due to dilution during preparation of the infusion . The covariates that reduced the variance of clearance from plasma and the volume of distribution by more than 10% were postnatal age (43 and 28%, respectively) and body weight (30.4 and 17.4%, respectively) . The expected reduction of clearance was about 10% for the plasma creatinine concentration . The other covariates studied (Apgar scores, plasma urea concentration, gestational age, sex) were found to have little effect . Simulations showed that a smaller percentage of patients had a maximum concentration in plasma/MIC ratio greater than 8 with a regimen of 7.5 mg/kg of body weight twice daily than with a regimen of 15 mg/kg once a day for MICs of 1 to 8 mg/liter.

Antimicrob Agents Chemother, 2002 May, 46(5), 1352 - 6
In vitro and in vivo activities of posaconazole against Coccidioides immitis; Gonzalez GM et al.; Posaconazole (SCH 56592) was tested against 25 strains of Coccidioides immitis to determine their in vitro susceptibilities . The geometric mean 48-h MIC of posaconazole (POSA) was 0.5 microg/ml, the MIC range was 0.25 to 1 microg/ml, and the MIC at which 50% of the isolates tested are inhibited (MIC50) and the MIC90 were 0.5 and 1 microg/ml, respectively . The geometric mean 48-h MIC of itraconazole (ITRA) was 0.23 microg/ml, the MIC range was 0.125 to 0.5 microg/ml, and the MIC50 and MIC90 were both 0.25 microg/ml . Two strains of C . immitis were selected for in vivo studies on the basis of the POSA 48-h MICs for the isolates . POSA orally administered at 0.01, 0.1, 0.5, 1, 5, and 10 mg/kg of body weight/day was compared with ITRA administered at 10 and 30 mg/kg three times a day . The spleens and livers of mice that died or survived to day 50 were removed to measure the fungal burdens . Mice had >or=90% survival when they were treated with >or=0.5 mg of POSA per kg or 30 mg of ITRA per kg . Cultures of whole spleens and livers from mice treated with 10 mg of POSA per kg showed >or=70% sterilization . No sterilization of whole spleens and livers from mice treated with ITRA was seen . POSA displayed potent in vivo activity against the two strains of C . immitis tested.

Antimicrob Agents Chemother, 2002 May, 46(5), 1325 - 8
Change in colony morphology of Candida lusitaniae in association with development of amphotericin B resistance; McClenny NB et al.; It is not uncommon to see amphotericin B treatment failure in patients with systemic infection caused by Candida lusitaniae . We report a patient with stage IV ovarian carcinoma and C . lusitaniae sepsis whose treatment with amphotericin B failed . The initial blood isolate was susceptible to amphotericin B in vitro; however, the MIC for a blood isolate recovered 7 weeks after treatment began showed a fourfold increase . Direct subculture of two positive blood samples obtained within a week of the patient's death showed the coexistence of two distinct colony color variants on CHROMagar Candida (CAC) . One variant was susceptible to amphotericin B, and one was resistant . These results emphasize the importance of repeat amphotericin B susceptibility testing for patients with persistent C . lusitaniae infection . The presence of colony variants on CAC may signal the emergence of amphotericin B resistance in C . lusitaniae and should be investigated.

J Infect Chemother, 2002 Mar, 8(1), 115 - 7
In-vitro activity of moxifloxacin and other fluoroquinolones against Chlamydia species; Miyashita N et al.; The in-vitro activity of moxifloxacin, a new fluoroquinolone, against Chlamydia species was investigated . The minimal inhibitory concentration of moxifloxacin for 10 standard strains of different Chlamydia species and 15 wild-type strains of Chlamydia pneumoniae isolated in Japan, which were morphologically different from clinical isolates from the United States, ranged from 0.031 to 0.125 microg/ml . The activity of moxifloxacin was almost the same as those of sparfloxacin, and it was 16, 8, 2, and 2 times more active than ciprofloxacin, levofloxacin, grepafloxacin, and tosufloxacin, respectively . The minimal chlamydiacidal concentration of moxifloxacin ranged from 0.031 to 0.125 microg/ml . These results suggest that moxifloxacin has potential effects against Chlamydia species.

Mol Immunol, 2002 May, 38(14), 1029 - 37
Asymmetric ligand recognition by the activating natural killer cell receptor NKG2D, a symmetric homodimer; Strong RK; Natural killer (NK) cells function through a diverse array of cell-surface natural killer receptors (NCRs) . NCRs specific for classical and non-classical MHC class I proteins, expressed in complex patterns of inhibitory and activating isoforms on overlapping, but distinct, subsets of NK cells, play an important role in immunosurveillance against cells that have reduced MHC class I expression as a result of infection or transformation . Another NCR, NKG2D, is an activating NCR first identified on NK cells, but subsequently found on macrophages and a variety of T cell types . NKG2D ligands in rodents include the MHC class I-like proteins RAE-1 and H60 and, in humans, ULBPs and the cell stress-inducible proteins MICA and MICB . NKG2D-MIC and -RAE-1 recognition events have been implicated in anti-viral and -tumor immune responses . Crystallographic analyses of NKG2D-MICA and -RAE-1 complexes reveal an unusual mode of recognition that apparently tolerates a surprising degree of ligand plasticity while generating affinities that are among the strongest TCR- or NCR-ligand affinities, thus, far described.

J Microbiol Methods, 2002 Jun, 50(1), 85 - 90
Non-radioactive PCR-SSCP with a single PCR step for detection of inhibitor resistant beta-lactamases in Escherichia coli; Alonso R et al.; A method based on PCR-SSCP has been developed to detect presumptive Inhibitor-Resistant TEM (IRT) beta-lactamases in Escherichia coli . The capacity of this technique to differentiate genes from 11 control strains encoding IRT beta-lactamases was evaluated with PCR products digested with RsaI . All the bla(TEM) genes studied could be distinguished by their electrophoretic mobilities . Applied to 29 epidemiologically unrelated clinical isolates of E . coli resistant to amoxicillin-clavulanate (MIC, > or =32 microg/ml), the electrophoretic mobilities of the digested bla(TEM) PCR products were identical to those of the reference bla(TEM-1A) (6 strains) and bla(TEM-1B) (18 strains) genes . The remaining five bla(TEM) PCR products displayed SSCP profiles different from those of the reference bla(TEM) genes and their nucleotide sequence identified them as bla(TEM-1C) in one strain, bla(TEM-30/IRT-2) in two strains, bla(TEM-37/IRT-8) in one strain, and bla(TEM-40/IRT-11) in one isolate . Overexpression of the wild-type bla(TEM-1) gene, as detected by high-level resistance to beta-lactams and enzyme assay, accounted for resistance in the 24 E . coli containing bla(TEM-1) . We report a simple one PCR step SSCP that can be used in epidemiological studies for rapid preliminary detection of IRT beta-lactamases; identification should be confirmed by sequence data.

Microbiol Immunol, 2002, 46(2), 89 - 93
Antifungal pharmacodynamic characteristics of amphotericin B against Trichosporon asahii, using time-kill methodology; Toriumi Y et al.; We determined the MIC of amphotericin B against 45 Trichosporon asahii isolates from various clinical and environmental sources, and used in vitro time-kill methods to characterize the relationship between amphotericin B concentrations and MIC for four representative T . asahii isolates . Amphotericin B had concentration-dependent antifungal activity . MICs ranged from 0.5 to 16 microg/ml, and most T . asahii isolates (76%, 34/45) were inhibited at safely achievable amphotericin B serum concentrations (< or = 2 microg/ml) . However, 40% (18/45) of isolates were not killed at these concentrations (MFCs from 1.0 to 32 microg/ml) . At concentrations > or = 2 x MIC, amphotericin B exhibited fungicidal activity (< 99.9% reduction in CFU) over a 12-hr time-period; the maximal effect was achieved at > or =4 x MIC . Susceptibility testing confirmed the resistance of T . asahii to amphotericin B, and in vitro pharmacodynamic results also suggest that amphotericin B is not suitable therapy for T . asahii infection.

Yao Xue Xue Bao, 1998, 33(3), 171 - 4
{Protective effects of (-)-S.R-daurisoline on neuronal injury in rat primary cortical cultures}; Liu J et al.; The neuroprotective effects of (-)-S.R-daurisoline on glutamate-induced neurotoxicity were studied in rat primary cortical cultures . The inhibitory effects of (-)-S.R-daurisoline on glutamate-elicited free intracellular Ca2+ increase were also studied in freshly dissociated single brain cells isolated from new born rat using AR-CM-MIC Cation Measurement System . Our experimental results demonstrated that (-)-S.R-daurisoline could obviously inhibit the neurotoxicity induced by glutamate and significantly increased cell viability in dose-dependent manner . In inhibiting glutamate-induced neurotoxicity, the IC50 value of (-)-S.R-daurisoline was found to be 3.4 mumol.L-1 . (-)-S.R-daurisoline was also shown to markedly inhibit glutamate-elicited increase of cytosolic free Ca2+ concentration in dose-dependent manner with IC50 value of 2.0 mumol.L-1 . Our results showed that (-)-S.R-daurisoline has an obvious protective effect on glutamate-induced neurotoxicity in primary cortical cultures . The protective mechanism of (-)-S.R-daurisoline may be relevant to inhibit Ca2+ influx into cells via glutamate-mediated ligand-gated ion channels.

Yao Xue Xue Bao, 1998, 33(3), 165 - 70
{Effects of daurisoline and its three optical isomers on ischemic injury in cultured pheochromocytoma (PC12) cells}; Liu J et al.; In this study, the protective effects of (-)-R.R-daurisoline and its three optical isomers on ischemic injury in cultured PC12 cells induced by treating cells with NaCN in glucose-free medium were investigated . Cell viability was measured using MTT assay . The results indicated that these compounds, especially (-)-S.R and (+)-R.S isomers were found substantially to attenuate ischemic injury in PC12 cells in a dose-dependent manner . The IC50 values of (-)-R.R, (-)-S.R, (+)-R.S and (+)-S.S isomers were shown to be 18.6 x 10(-6), 2.4 x 10(-6), 5.9 x 10(-6) and 90 x 10(-6) mol.L-1, respectively . Intracellular free Ca2+ concentration in PC12 cells was measured using AR-CM-MIC cation measurement system with Fura-2/AM as Ca2+ fluorescent indicator . (-)-R.R-daurisoline and its three optical isomers: (-)-S.R, (+)-R.S and (-)-S.S were found to markedly inhibit the increase of cytosolic free Ca2+ concentration induced by NaCN (20 mmol.L-1) in a dose-dependent manner . Their IC50 were found to be 3.55 x 10(-6), 0.59 x 10(-6), 1.29 x 10(-6) and 24.3 x 10(-6) mol.L-1 respectively . It is suggested that the cytoprotective effects of daurisoline and its isomers were mediated by blocking Ca2+ influx into cells.

J Clin Pharmacol, 2002 Apr, 42(4), 403 - 11
Comparative target site pharmacokinetics of immediate- and modified-release formulations of cefaclor in humans; de PA et al.; Optimal dosing of beta-lactam antibiotics aims at maximizing the time at which drug levels in the interstitial space fluid (ISF)--the fluid that surrounds the causative microorganisms at the target site--exceed the minimal inhibitory concentration (MIC) . One potentially attractive strategy to achieve this goal is to administer antibiotics as oral sustained-release formulations . The present study was designed to test the hypothesis that sustained-release formulations could lead to a more suitable pharmacokinetic profile in the ISF at the relevant target site . For this purpose, time versus cefaclor concentration profiles attained in the ISF were measured following administration of two formulations, an immediate- (500 mg IR) and a modified-release formulation in two different doses (500 mg MR and 750 mgMR) in a three-way crossover study of healthy male volunteers (n = 12) . For the measurement of unbound cefaclor concentrations in the ISF of human skeletal muscle, the in vivo microdialysis technique was employed . For all three formulations, unbound cefaclor concentration in the ISF closely followed individual plasma concentration profiles in a dose-dependent pattern, with ISF to unbound plasma ratios ranging from 0.67 to 0.73 . The mean residence time was found to be significantly longer for the MR formulations versus the IR formulation . The data of the present study indicate that time above MIC values at the target site can be substantially prolonged if an antibiotic is administered as a sustained-release product.

Phytother Res, 2002 Mar, 16 Suppl 1, S89 - 90
Causal organism of flacherie in the silkworm Antheraea assama Ww: isolation, characterization and its inhibition by garlic extract; Choudhury A et al.; Of the different bacterial strains isolated from diseased muga silkworms, the strain named as AC-3 was found to be most pathogenic to the silkworm . Different antibiotics and plant extracts were tested for their effectiveness in inhibiting the growth of AC-3 . Fresh Allium sativum (garlic extract) was most effective against the strain . The stability and MIC of the garlic extract has also been studied . We report for the first time the effectiveness of garlic extract in controlling the bacterium causing disease in the muga silkworm .

Int J Antimicrob Agents, 2002 Mar, 19(3), 183 - 7
Effects of gatifloxacin on phagocytosis, intracellular killing and oxidant radical production by human polymorphonuclear neutrophils; Braga PC et al.; The ingestion and killing of bacteria by phagocytic cells is an important step in the sequence of interactions between invading microorganisms and host defense systems and may be affected by antibiotics . We investigated the effects of gatifloxacin on the phagocytosis, killing and oxidative bursts of human polymorphonuclear neutrophils (PMNs) . The percentage phagocytosis and the phagocytosis index were unaffected by exposure of Escherichia coli strains to sub-MICs of gatifloxacin to a 1/64 dilution . However a significant increase in percentage intraphagocytic killing and the killing index occurred in one E . coli strain at 1/32 MIC and in two strains at 1/16 MIC . The incubation of PMNs with sub-MICs and supra-MICs of gatifloxacin (to 32 MIC) did not affect the oxidative bursts.

Int J Tuberc Lung Dis, 2002 Feb, 6(2), 164 - 5
Investigation of cross-resistance between rifampin and rifabutin in Mycobacterium tuberculosis complex strains; Uzun M et al.; In order to indicate the cross-resistance between rifampin (RMP) and rifabutin (RBU), minimal inhibitory concentrations (MICs) of RBU were investigated in 50 Mycobacterium tuberculosis strains . The MIC values of 25 RMP-susceptible (to 2 microg/ml) and 25 RMP-resistant (to 2 microg/ml) M . tuberculosis strains against RBU were determined by the Bactec TB 460 system . All of the RMP-susceptible strains were also susceptible to RBU (MIC < or = 1 microg/ml) . Three out of 25 (12%) RMP-resistant strains were determined as susceptible to RBU . The high level cross-resistance (88%) obtained in this study highlights the importance of testing susceptibility to RBU prior to its inclusion in the tuberculosis treatment regimens at the Istanbul Faculty of Medicine.

Folia Med (Plovdiv), 2001, 43(3), 10 - 2
Drug susceptibility testing of Malassezia furfur strains to antifungal agents; Zissova LG et al.; The minimal inhibitory concentration of the three most commonly used antifungal agents--ketoconazole, fluconazole and intraconazole against 28 strains of Malassezia furfur, was studied . The strains were recovered from patients with pityriasis versicolor, pityriasis simplex capitis and dermatitis seborrhoides and were cultured on Dixon's agar . Diagnostic sensitivity test agar was used to determine the minimal inhibitory concentrations of the antifungal agents . The established minimal inhibitory concentration of ketoconazole against Malassezia furfur was 0.02 mg/l, the minimal inhibitory concentration of intraconazole--0.05 mg/l and that of fluconazole--0.09 mg/l . The results of the study are consistent with the data reported by other European authors in their studies . On the basis of the accumulated data it is concluded that the tested antifungal agents are effective in the treatment of infections caused by Malassezia furfur but ketoconazole is superior to intraconazole and fluconazole in the treatment of these infections . This is the first study on the drug susceptibility of Malassezia furfur in Bulgaria.

Lijec Vjesn, 2001 Nov-Dec, 123(11-12), 293 - 6
{Effect of inoculum size on sensitivity and specificity of the double-disk synergy test for the detection of wide-spectrum beta-lactamases}; Bedenic B et al.; The plasmid-mediated extended-spectrum beta-lactamases (ESBL) confer resistance to oxymino-cephalosporins, such as cefotaxime, ceftazidime, and ceftriaxone and to monobactams such as aztreonam . It is well known fact that ESBL producing bacteria exhibit a pronounced inoculum effect against broad spectrum cephalosporins like ceftazidime, cefotaxime, ceftriaxone and cefoperazone . The aim of this investigation was to determine the effect of inoculum size on the sensitivity and specificity of double-disk synergy test (DDST) which is the test most frequently used for detection of ESBLs, in comparison with other two methods (determination of ceftazidime MIC with and without clavulanate and inhibitor potentiated disk-diffusion test) which are seldom used in clinical laboratories . The experiments were performed on a set of K . pneumoniae strains with previously characterized beta-lactamases which comprise: 10 SHV-5 beta-lactamase producing K . pneumoniae, 20 SHV-2 + 1 SHV 2a beta-lactamase producing K . pneumoniae, 7 SHV-12 beta-lactamase producing K . pneumoniae, 39 putative SHV ESBL producing K . pneumoniae and 26 K . pneumoniae isolates highly susceptible to ceftazidime according to Kirby-Bauer disk-diffusion method and thus considered to be ESBL negative . According to the results of this investigation, increase in inoculum size affected more significantly the sensitivity of DDST than of other two methods . The sensitivity of the DDST was lower when a higher inoculum size of 10(8) CFU/ml was applied, in distinction from other two methods (MIC determination and inhibitor potentiated disk-diffusion test) which retained high sensitivity regardless of the density of bacterial suspension . On the other hand, DDST displayed higher specificity compared to other two methods regardless of the inoculum size . This investigation found that DDST is a reliable method but it is important to standardize the inoculum size.

Toxicol Appl Pharmacol, 2002 Apr 1, 180(1), 22 - 35
Antitumor effects of miconazole on human colon carcinoma xenografts in nude mice through induction of apoptosis and G0/G1 cell cycle arrest; Wu CH et al.; Miconazole (MIC), a promising oral antifungal agent, has been used worldwide in the treatment of superficial mycosis . In this study, we demonstrated that MIC dose dependently arrested various human cancer cells at the G0/G1 phase of the cell cycle . The protein levels of p53, p21/Cip1, and p27/Kip1 were significantly elevated by MIC treatment in COLO 205 cells . Electrophoretic mobility gel shift assays showed that the nuclear extracts of the MIC-treated COLO 205 cells exerted a significant binding between wild-type p53 and its consensus-binding site present in the p21/Cip1 promoter . These results suggested that the p53-associated signaling pathway is involved in the regulation of MIC-induced cancer cell growth arrest . By immunoblot analysis, we demonstrated that cyclin D3 and cyclin-dependent kinase-4 (CDK4) protein levels were inhibited by MIC treatment in the cancer cells . Significant therapeutic effect was further demonstrated in vivo by treating nude mice bearing COLO 205 tumor xenografts with MIC (50 mg/kg ip) . The protein expression of p53 was significantly increased in MIC-treated tumor tissues by immunohistochemical staining and Western blotting analysis . DNA fragmentation and TUNEL assay were performed and demonstrated that apoptosis occurred in tumor tissues treated with MIC . Our study provides the novel mechanisms of antitumor effects of MIC and such results may have significant applications for cancer chemotherapy . (c)2002 Elsevier Science (USA).

Avian Dis, 2002 Jan-Mar, 46(1), 215 - 8
In vitro antibiotic susceptibility of field isolates of Mycoplasma synoviae in Argentina; Cerda RO et al.; Minimum inhibitory concentrations (MICs) were determined in vitro for 7 antibiotics (aivlosin, enrofloxacine, tylosin, tiamulin, kitasamycin, chlortetracycline, and oxytetracycline) against eight recent local Argentinean isolates and two standard strains of Mycoplasma synoviae . Aivlosin (3-acetyl-4"-isovaleryl tylosin tartrate), tylosin, and tiamulin showed the lowest MICs with MIC90s of 0.006, 0.012, and 0.05 microg/ml, respectively . Except one strain that showed resistant values to chlortetracycline (> or = 12.5 microg/ml), all the analyzed strains were susceptible in different degrees to all the antibiotics tested . In this study, the improved activity of the tylosin-derived drug, aivlosin, was confirmed because it showed, in most strains, MIC values half those for tylosin.

J Clin Microbiol, 2002 Apr, 40(4), 1406 - 12
In vitro activity of nystatin compared with those of liposomal nystatin, amphotericin B, and fluconazole against clinical Candida isolates; Arikan S et al.; We investigated the in vitro activity of nystatin and liposomal nystatin against 103 Candida isolates to determine the effect of both time and medium on MICs . We also compared the nystatin MICs with those of amphotericin B and fluconazole . Testing was performed in accordance with the National Committee for Clinical Laboratory Standards M27-A microdilution methodology with RPMI 1640, RPMI 1640 supplemented with glucose to 2% (RPMI-2), and antibiotic medium 3 supplemented with glucose to 2% (AM3) . While nystatin MICs were similar to or slightly lower than liposomal nystatin MICs in RPMI 1640 and RPMI-2, they were markedly higher than liposomal nystatin MICs in AM3 . Use of AM3 and determination of the MIC after 24 h of incubation provided a slightly wider range of liposomal nystatin MICs (0.06 to >16 microg/ml) . Under these conditions, the MICs at which 90% of isolates were inhibited of nystatin and liposomal nystatin were 2 and 1 microg/ml, respectively . Nystatin and liposomal nystatin in general showed good activity against all Candida spp . tested . Although the MICs of nystatin and liposomal nystatin tended to rise in parallel with the amphotericin B MICs, nystatin and liposomal nystatin MICs of 1 to 2 and 0.5 to 1 microg/ml, respectively, were obtained for seven and six, respectively, of nine isolates for which amphotericin B MICs were >or=0.25 microg/ml . No correlation between fluconazole and nystatin or liposomal nystatin MICs was observed . As amphotericin B MICs of >or=0.25 microg/ml correlate with in vitro resistance, these results suggest that liposomal nystatin might have activity against some amphotericin B-resistant isolates . In vivo testing in animal models is required for clarification of this issue.

J Invest Dermatol, 2002 Apr, 118(4), 686 - 91
MICA gene polymorphism is not associated with an increased risk for skin cancer; Kennedy C et al.; The MICA gene encodes for major histocompatibility complex class I chain-related proteins (MIC), which belong to a recently identified new family of nonclassical major histocompatibility complex molecules . The general structure of the MICA molecule resembles that of major histocompatibility complex class I molecules . MIC molecules are considered to be stress-induced antigens that are recognized by cytotoxic T cells and natural killer cells, which play an important role in the surveillance of transformed infected and damaged cells . Associations of major histocompatibility complex class I molecules with skin cancer have been described before . To evaluate the possible association of MICA gene polymorphism with the risk for nonmelanoma skin cancer we evaluated 153 cases with squamous cell carcinoma, 261 cases with basal cell carcinoma, 111 controls with malignant melanoma, and 247 controls without a history of skin cancer . Five distinct MICA alleles A4, A5, A6, A9, and A5.1 were studied . As the MICA 5.1 variant gene contains a four-nucleotide insertion that causes a stop codon in the trans membrane region, the resulting truncated MICA molecule does not reside on the cellular membrane . In the case of individuals who are homozygous for MICA 5.1 this results in cells that are naked for the MICA molecule . We therefore specifically addressed the possible association between MICA 5.1 homozygosity and skin cancer, as these individuals are expected to be at the highest risk for skin cancer if the MICA gene plays a role in skin carcinogenesis . Viral proteins may serve as antigens for recognition of skin cancer by the immune system . Human papillomavirus is the most likely candidate virus to be involved in the carcinogenesis of cutaneous squamous cell carcinoma . Hence, we also assessed the association between MICA polymorphism and squamous cell carcinoma in human-papillomavirus-positive and human-papillomavirus-negative individuals as identified by the presence of human papillomavirus DNA in hairs plucked from their eyebrows . Our analyses did not reveal any significant differences regarding the MICA allele frequencies between cases and controls . Also homozygotes and heterozygotes for the MICA 5.1 variant gene were not at an increased risk for skin cancer compared to individuals without this variant gene and infection with human papillomavirus did not materially influence these findings . The same group of cases and controls was large enough to show an association between melanocortin 1 receptor gene polymorphism and skin cancer and to reasonably exclude an association between p53 codon 72 polymorphism and skin cancer . Therefore, we conclude that an association between MICA gene polymorphism and nonmelanoma skin cancer is not likely.

J Invest Dermatol, 2002 Apr, 118(4), 600 - 5
Expression of stress-induced MHC class I related chain molecules on human melanoma; Vetter CS et al.; Cellular immune responses to melanoma are tightly regulated and include specific T cell responses to self antigens such as Mart-1 and gp100 . Thus, additional signals apart from those mediated by the T cell receptor are needed to ensure T cell activation . Recently, the stress inducible major histocompatibility complex molecules, MHC class I related chain, were identified as an activator of both natural killer and T cells via interaction with their receptor NKG2D . Herein, we report the expression of MIC in 31 of 40 primary cutaneous melanomas and in 13 of 20 metastatic lesions . Moreover, lymphocytes infiltrating the tumor were found to express NKG2D . Detailed analysis identified both CD3+ T cells as well as CD56+ natural killer cells contributing to this NKG2D+ tumor infiltrating lymphocyte population present.

J Antibiot (Tokyo), 2002 Jan, 55(1), 30 - 5
YM-181741, a novel benz{a}anthraquinone antibiotic with anti-Helicobacter pylori activity from Streptomyces sp; Taniguchi M et al.; A novel benz{a}anthraquinone, YM-181741, was isolated from the culture broth of actinomycete strain Q57219 . The strain was identified as Streptomyces sp . by morphological and chemotaxonomic characterization . YM-181741 was purified from the culture supernatant by serial column chromatography . The structure of YM-181741 was determined by spectroscopic analysis including one- and two-dimensional NMR experiments . YM-181741 showed selective anti-Helicobacterpylori activity with a MIC value of 0.2 microg/ml.

J Bacteriol, 2002 Apr, 184(8), 2131 - 40
Mutations in the 16S rRNA genes of Helicobacter pylori mediate resistance to tetracycline; Trieber CA et al.; Low-cost and rescue treatments for Helicobacter pylori infections involve combinations of several drugs including tetracycline . Resistance to tetracycline has recently emerged in H . pylori . The 16S rRNA gene sequences of two tetracycline-resistant clinical isolates (MIC = 64 microg/ml) were determined and compared to the consensus H . pylori 16S rRNA sequence . One isolate had four nucleotide substitutions, and the other had four substitutions and two deletions . Natural transformation with the 16S rRNA genes from the resistant organisms conferred tetracycline resistance on susceptible strains . 16S rRNA genes containing the individual mutations were constructed and tested for the ability to confer resistance . Only the 16S rRNA gene containing the triple mutation, AGA965-967TTC, was able to confer tetracycline resistance on H . pylori 26695 . The MICs of tetracycline for the transformed strains were equivalent to those for the original clinical isolates . The two original isolates were also metronidazole resistant, but this trait was not linked to the tetracycline resistance phenotype . Serial passage of several H . pylori strains on increasing concentrations of tetracycline yielded mutants with only a very modest increase in tetracycline resistance to a MIC of 4 to 8 microg/ml . These mutants all had a deletion of G942 in the 16S rRNA genes . The mutations in the 16S rRNA are clearly responsible for tetracycline resistance in H . pylori.

Br J Ophthalmol, 2002 Apr, 86(4), 387 - 9
Vitreous penetration of levofloxacin in the uninflamed phakic human eye; Herbert EN et al.; AIMS: To assess the vitreous penetration of oral levofloxacin (a new fluoroquinolone antibiotic with improved Gram positive activity) in uninflamed phakic eyes . METHODS: 15 patients for macula hole surgery were recruited to the study . 10 received a single 500 mg dose of levofloxacin by mouth preoperatively . Five acted as controls . Serum and undiluted vitreous samples were obtained at surgery and analysed by HPLC . RESULTS: Levofloxacin was detectable 2.5 hours after administration in the vitreous . A peak concentration of 1.6 microg/ml (or mg/l) was measured between 2.5 and 4 hours post-dose . CONCLUSION: Oral levofloxacin reaches the vitreous rapidly in the uninflamed phakic eye . Levels did not reach MIC(90) for the commonest infecting organisms . Nevertheless, levofloxacin would be expected to be active against a higher proportion of infecting organisms than either ciprofloxacin or ofloxacin.

J Antimicrob Chemother, 2002 Apr, 49(4), 679 - 81
Comparison of phenotypic and genotypic methods for the detection of clarithromycin resistance in Mycobacterium avium; Thiermann S et al.; The MICs of clarithromycin for 10 clinical isolates of Mycobacterium avium were determined using three methods: Bactec 460-TB, broth microdilution and Etest . The results were compared with the presence of resistance mutations in the 23S rRNA gene . Isolates were obtained from five AIDS patients who were treated with clarithromycin . Five isolates were recovered before and five during treatment . MICs were reproducible and comparable between the three methods . They were < or = 4 mg/L for pre-treatment isolates and > or = 128 mg/L for strains recovered during treatment . An MIC > or = 128 mg/L was associated with the presence of mutations in the 23S rRNA gene that were absent in the isolates exhibiting MIC < ro = 4 mg/L.

J Nat Prod, 2002 Mar, 65(3), 249 - 54
(+)-7S-Hydroxyxestospongin A from the marine sponge Xestospongia sp . and absolute configuration of (+)-xestospongin D; Moon SS et al.; The structure of the title compound, (+)-7S-hydroxyxestospongin A was solved by single-crystal X-ray diffraction analysis and the absolute stereochemistry obtained by analysis of the derived R and S Mosher's esters . The absolute configuration of xestospongin D was determined for the first time by analysis of anomalous scattering from the X-ray crystal diffraction data set . Xestospongins A, C, and D, araguspongine C, and demethylxestospongin B exhibited modest antifungal activity (MIC 30-100 g/mL) against various fluconazole-resistant Candida spp., but 7S-hydroxyxestospongin A was inactive.

Histopathology, 2001 Dec, 39(6), 578 - 83
CD99/MIC-2 surface protein expression in breast carcinomas; Milanezi F et al.; AIM: To evaluate the expression of CD99/MIC-2 surface protein in invasive breast carcinomas and demonstrate whether or not there is a relationship with tumour phenotype . METHODS AND RESULTS: Thirty-five invasive breast carcinomas, including five metaplastic carcinomas, were stained with CD99 primary antibodies using standard protocols based on streptavidin-biotin-peroxidase method . Four out of five metaplastic carcinomas expressed CD99/MIC-2 protein, three of them were matrix-producing carcinomas . From the other 30 cases, only an invasive apocrine carcinoma was positive . There was no statistical correlation between CD99 expression and the parameters analysed (histological typing and grading, proliferative index and nodal status) . CONCLUSIONS: CD99/MIC-2 is expressed in breast carcinomas, especially in the matrix-producing variant of metaplastic carcinomas, which impairs its use as a marker to differentiate metaplastic carcinomas from primary and metastatic sarcomas of the breast . It seems to have no prognostic implications . However, phenotype similarities with other chondromyxoid tumours that also express the protein, like mesenchymal chondrosarcomas, suggest a relationship between MIC-2 reactivity and morphological differentiation.

Pathology, 2002 Feb, 34(1), 71 - 3
Increasing resistance of Helicobacter pylori to clarithromycin: is the horse bolting?
Grove DI, Koutsouridis G.
AIMS: To determine whether there has been a change in the patterns of susceptibility to various antibiotics of our isolates of Helicobacter pylori over a 5-year period from 1996 to 2000 . METHODS: Five hundred and fourteen isolates of H . pylori grown from gastric biopsies were tested for susceptibility to amoxycillin, clarithromycin, metronidazole and tetracycline . The usage of macrolide antibiotics in Australia was examined by calculating the numbers of prescriptions issued under the Australian pharmaceutical benefits scheme between 1992 and 2000 . RESULTS: There were no changes in susceptibility of H . pylori to amoxycillin and tetracycline and there was a slight decline in resistance to metronidazole . In contrast, there was a stepwise 4-fold increase from 3.8 to 15.7% in the number of isolates resistant to clarithromycin and a similar increase in the mean minimum inhibitory concentration of clarithromycin during the 5-year period of observation . There was no change in overall macrolide consumption in Australia over this and the preceding 3 years . However, the pattern changed, with erythromycin usage being halved and being replaced by roxithromycin and clarithromycin . CONCLUSIONS: Resistance of H . pylori to clarithromycin is increasing, possibly as a consequence of increased usage of roxithromycin and clarithromycin . More patients are likely to fail to respond to empirical therapy and will need microbiological investigation.

Chemotherapy, 2002 Mar, 48(1), 21 - 5
Comparison of the Sensititre YeastOne colorimetric microdilution panel and the NCCLS broth microdilution method for antifungal susceptibility testing against Candida species; Bernal S et al.; We evaluated the commercially prepared Sensititre YeastOne colorimetric antifungal panel to determine the susceptibility of 170 Candida spp isolates to amphotericin B, fluconazole, itraconazole, and flucytosine . The NCCLS reference microdilution method (M27-A document) was used as reference method . The YeastOne panel was performed according to the manufacturer's instructions . For the colorimetric method, MICs were determined at 24 h of incubation . MICs for the NCCLS reference method were read at 48 h of incubation . The overall agreement within +/-2 dilutions by both methods was calculated against the four antifungal agents . This agreement was 92.9, 68.2, 77.6 and 80% for amphotericin B, fluconazole, itraconazole, and flucytosine, respectively . Thirteen isolates (7.6%) showed very major discrepancies for fluconazole and 12 (7%) for itraconazole . We found that the reading of MIC with the YeastOne panel was somewhat easier than the reading of reference MIC, although the determination of endpoint was sometimes difficult, especially for azoles, because the trailing effect appeared in a high percentage of isolates .

Antimicrob Agents Chemother, 2002 Apr, 46(4), 1144 - 6
In vitro antifungal susceptibilities of Trichosporon species; Paphitou NI et al.; The in vitro activities of amphotericin B, itraconazole, fluconazole, voriconazole, posaconazole, and ravuconazole against 39 isolates of Trichosporon spp . were determined by the NCCLS M27-A microdilution method . The azoles tested appeared to be more potent than amphotericin B . Low minimal fungicidal concentration/MIC ratios were observed for voriconazole, posaconazole, and ravuconazole, suggesting fungicidal activity.

Pancreas, 2002 Apr, 24(3), 264 - 8
Penetration of meropenem and cefepim into pancreatic tissue during the course of experimental acute pancreatitis; Saglamkaya U et al.; INTRODUCTION: Recent data from experimental and clinical studies suggest that the antibiotics showing good penetration into the pancreas may reduce mortality by preventing pancreatic infection, which is the most important prognostic factor in acute pancreatitis . AIM: To determine and compare pancreatic tissue concentrations of meropenem and cefepime at different stages of acute necrotizing pancreatitis in an animal model that has been shown to closely mimic severe human pancreatitis . METHODOLOGY: Acute necrotizing pancreatitis was induced in rats by a standardized intraductal infusion of glycodeoxycholic acid and intravenous cerulein . Six hours (n = 30) and 48 hours (n = 30) after induction of pancreatitis, the rats were randomized to receive an intravenous 20 mg/kg injection of either meropenem or cefepime . Blood and the head of the pancreas were collected for determining antibiotic concentrations by high-performance liquid chromatography . RESULTS: Meropenem concentrations in the pancreas at 6 hours of acute pancreatitis increased significantly and decreased at 48 hours of the disease, but were still higher than that in controls . Concentrations of cefepime in necrotic pancreatic tissue were significantly low either during the initial or later phase, but lower in latter, in which the necrosis was more evident . Tissue/serum concentration ratios of meropenem were significantly higher than those of cefepime . However, tissue concentrations of both antibiotics are much higher than the minimum inhibitory concentration values for the common microorganisms involved in pancreatic infections . CONCLUSION: Although both antibiotics penetrate into the necrotic tissue in sufficient therapeutic concentrations, penetration of meropenem is much better than cefepime . However, good tissue penetration may not solely indicate efficacy of that antibiotic . Therefore, further experimental and clinical studies are needed to determine the therapeutic and prognostic efficacy of these agents.

Pflugers Arch, 2002 Mar, 443(5-6), 882 - 91 Epub 2002 Jan 22.
Influence of voltage and extracellular Na(+) on amiloride block and transport kinetics of rat epithelial Na(+) channel expressed in Xenopus oocytes; Segal A et al.; We expressed the three subunits of the epithelial amiloride-sensitive Na(+) channel (ENaC) from rat distal colon heterologously in oocytes of Xenopus laevis and analysed blocker-induced fluctuations in current using conventional dual-microelectrode voltage-clamp . To minimize Na(+) accumulation we performed all experiments in low-Na(+) solutions (15 mM) . Noise analysis revealed that control or ENaC-injected oocytes did not exhibit spontaneous relaxation noise . However, in ENaC-expressing oocytes, amiloride induced a distinct Lorentzian component in the power density spectra . With three amiloride concentrations and a linear analysis of the respective changes in the corner frequency f(c) (2 pi f(c) plot) we determined the rate constants k(on) and k(off) for the amiloride-ENaC interaction . At a clamp potential (V(m)) of -60 mV k(on) was 80.8 +/- 5.1 microM(-1) s(-1) and k(off) 15.4 +/- 4.2 s(-1) . The half-maximal blocker concentration (K(mic,ami)) was 0.19 microM (V(m)=-60 mV) . While k(on) was voltage-independent in the range -50 to -100 mV, k(off) and K(mic,ami) decreased significantly with increasing membrane hyperpolarization, resulting in an increased affinity of amiloride for its binding site on ENaC . Increasing extracellular {Na(+)} ({Na(+)}(o)) led to saturation of ENaC . Subsequent noise analysis revealed that single-channel current increased non-linearly with {Na(+)}(o) and that saturation was not due to a reduction in the number of open channels . The apparent affinity of Na(+) for its binding site on the channel was voltage dependent and increased with hyperpolarization . Noise analysis revealed that k(on) and k(off) for amiloride decreased with increasing {Na(+)}(o), while the affinity of the amiloride-binding site did not change . These findings show that the affinity of rat intestinal ENaC for amiloride is voltage dependent and is influenced non-competitively by {Na(+)}(o), indicating that Na(+) and amiloride do not compete for the same binding site at the channel.

Plant Cell, 2002 Feb, 14(2), 333 - 42
Cloning of tangerine from tomato reveals a carotenoid isomerase essential for the production of beta-carotene and xanthophylls in plants; Isaacson T et al.; Carotenoid biosynthesis in plants has been described at the molecular level for most of the biochemical steps in the pathway . However, the cis-trans isomerization of carotenoids, which is known to occur in vivo, has remained a mystery since its discovery five decades ago . To elucidate the molecular mechanism of carotenoid isomerization, we have taken a genetic map-based approach to clone the tangerine locus from tomato . Fruit of tangerine are orange and accumulate prolycopene (7Z,9Z,7'Z,9'Z-tetra-cis-lycopene) instead of the all-trans-lycopene, which normally is synthesized in the wild type . Our data indicate that the tangerine gene, designated CRTISO, encodes an authentic carotenoid isomerase that is required during carotenoid desaturation . CRTISO is a redox-type enzyme structurally related to the bacterial-type phytoene desaturase CRTI . Two alleles of tangerine have been investigated . In tangerine(mic), loss of function is attributable to a deletion mutation in CRTISO, and in tangerine(3183), expression of this gene is impaired . CRTISO from tomato is expressed in all green tissues but is upregulated during fruit ripening and in flowers . The function of carotene isomerase in plants presumably is to enable carotenoid biosynthesis to occur in the dark and in nonphotosynthetic tissues.

J Immunol, 2002 Mar 15, 168(6), 2988 - 96
Lamina propria CD4+ T lymphocytes synergize with murine intestinal epithelial cells to enhance proinflammatory response against an intracellular pathogen; Mennechet FJ et al.; Acute and lethal ileitis can be elicited in certain strains of inbred mice after oral infection with the intracellular protozoan parasite Toxoplasma gondii . The development of this inflammatory process is dependent upon the induction of a robust Th1 response, including overproduction of IFN-gamma, TNF-alpha, and NO, as has been reported in other experimental models of human inflammatory bowel disease . In this study we have investigated the role of CD4(+) T cells from the lamina propria (LP) in the early inflammatory events after T . gondii infection using isolated and primary cultured intestinal cells from infected mice and immortalized mouse mIC(cl2) intestinal epithelial cells . Primed LP CD4(+) T cells isolated from parasite-infected mice produce substantial quantities of both IFN-gamma and TNF-alpha . IFN-gamma- and TNF-alpha-producing LP CD4(+) T cells synergize with infected mIC(cl2) and enhance the production of several inflammatory chemokines including macrophage-inflammatory protein-2, monocyte chemoattractant protein-1, monocyte chemoattractant protein-3, macrophage-inflammatory protein-1alphabeta, and IFN-gamma-inducible protein-10 . Furthermore, primed LP CD4(+) T cells cocultured with infected mIC(cl2) inhibited replication of the parasite in the intestinal epithelial cells . Thus, LP CD4(+) T cells can interact with parasite-infected intestinal epithelial cells and alter the expression of several proinflammatory products that have been associated with the development of intestinal inflammation . The interaction between these two components of the gut mucosal compartment (CD4(+) T cells and enterocytes) may play a role in the immunopathogenesis of this pathogen-driven experimental inflammatory bowel disease model.

Zhonghua Xue Ye Xue Za Zhi, 2001 Jun, 22(6), 313 - 5
{The Immunophenotypical features of t (8; 21) (q22; q22) acute myeloid leukemia}; Pan X et al.; OBJECTIVE: To study the predictive value of immunophenotypical features in t (8; 21) (q22; q22) acute myeloid leukemia (AML) . METHODS: Morphological/cytochemical, flow cytometric immunophenotyping, cytogenetic analyses (MIC) and RT-PCR were performed in 294 previously untreated AML . RESULTS: (1) In 294 AML patients, t (8; 21) AML were 21.8% (64); in AML-M(2), t (8; 21) AML were 54.7%; and in t (8; 21) AML, AML-M(2) were 81.3% . (2) Compared with control group, CD(19) and CD(34) expressions were higher, and CD(33) expression was lower (P < 0.001) in t (8; 21) AML . (3) If the cut-off value of CD(19) positive was >or= 20%, CD(19) positive rate was 13.6% (40/294) in AML, and 50% (32/64) and 3.5% (8/230) (P < 0.001) in t (8; 21) AML and control group . (4) CD(19)(+) and/or CD(34)(+) t (8; 21) AML accounted for 90.6% (58/64) of t (8; 21) AML and CD(19)(-)/CD(34)(-) for 9.6% (6/64) . CONCLUSION: In t (8; 21) AML, especially M(2)/t (8; 21), CD(19) and CD(34) expressions were high . CD(19) was one of predictive markers of t (8; 21) AML.

Zhonghua Xue Ye Xue Za Zhi, 2000 Jan, 21(1), 14 - 6
{Comparative study of clinical features of childhood and adult acute lymphoblastic leukemia}; Liu X et al.; OBJECTIVE: To analyze the outcome, the immunological and cytogenetic characteristics and the frequency of primary multi-drug resistance of childhood acute lymphoblastic leukemia (ALL) in our hospital during the past two years . METHOD: The complete remission (CR) rate of 154 newly diagnosed ALL patients were analysed and the biological features of the leukemic cells were explored by immunochemistry and cytogenetics . RESULTS: The CR rate in the ALL children is 94.1% which is significantly higher than that of adult ALL patients (67.8%) . Immunophenotype analysis showed that 18.5% of the children expressed myeloid antigens besides lymphoid antigens and 4.8% of them were Ph chromosome positive.Both the figures were lower than that of adult patients . CONCLUSION: Childhood ALL has distinct morphological, immunological and cytogenetic (MIC) features, which might contribute to the good prognosis of the patients.

Syst Appl Microbiol, 2001 Dec, 24(4), 597 - 609
Structure and functional analysis of the microbial community in an aerobic: anaerobic sequencing batch reactor (SBR) with no phosphorus removal; Kong YH et al.; The bacterial community of an aerobic:anaerobic non-P removing SBR biomass fed a mixture of acetate and glucose was analysed using several 16S rRNA based methods . Populations responsible for anaerobic glucose and acetate assimilation were determined with fluorescent in situ hybridization (FISH) in combination with microautoradiography (FISH/MAR) . At 'steady state' this community consisted of alpha-Proteobacteria (26%) and gamma-Proteobacteria (14%), mainly appearing as large cocci in tetrads (i.e . typical 'G-Bacteria') . Large numbers of low G+C bacteria (22%), and high G+C Gram-positive bacteria (29%) seen as small cocci in clusters or in sheets were also detected after FISH . DGGE fingerprinting of PCR amplified 16S rDNA fragments and subsequent cloning and sequencing of several of the major bands led to the identification of some of these populations . They included an organism 98% similar in its 16S rRNA sequence to Micropruina glycogenica, and ca . 76% of the high G+C bacteria responded to a probe MIC 184, designed against it . The rest responded to the KSB 531 probe designed against a high G+C clone sequence, sbr-gs28 reported in other similar systems . FISH analyses showed that both these high G+C populations were almost totally dominated by small clustered cocci . Only ca . 2% of cells were beta-Proteobacteria . None of the alpha- and gamma-Proteobacterial 'G-bacteria' responded to FISH probes designed for the 'G-Bacteria' Amaricoccus spp . or Defluvicoccus vanus . FISH/MAR revealed that not all the alpha-Proteobacterial 'G-Bacteria' could take up acetate or glucose anaerobically . Almost all of the gamma-Proteobacterial 'G-Bacteria' assimilated acetate anaerobically but not glucose, the low G+C clustered cocci only took up glucose, whereas the high G+C bacteria including M . glycogenica and the sbr-gs28 clone assimilated both acetate and glucose . All bacteria other than the low G+C small cocci and a few of the alpha-Proteobacteria accumulated PHB . The low G+C bacteria showing anaerobic glucose assimilation ability were considered responsible for the lactic acid produced anaerobically by this SBR biomass, and M . glycogenica for its high glycogen content.

J Antimicrob Chemother, 2002 Mar, 49(3), 549 - 52
Use of a rapid mismatch PCR method to detect gyrA and parC mutations in ciprofloxacin-resistant clinical isolates of Escherichia coli; Qiang YZ et al.; Four amino acid substitutions, two in GyrA and two in ParC subunits of DNA gyrase and topoisomerase IV, respectively, are commonly responsible for fluoroquinolone resistance in Escherichia coli . In this study, an economical and time-efficient mismatch amplification mutation assay (MAMA) PCR was developed to detect mutations in the chromosomal gyrA and parC genes causing these substitutions . One hundred and twenty-one clinical E . coli isolates were tested by this assay, and the results confirmed that accumulation of amino acid alterations in GyrA and ParC correlates closely with stepwise increases in the MIC of ciprofloxacin.

J Antimicrob Chemother, 2002 Mar, 49(3), 507 - 13
Experimental study of the efficacy of vancomycin, rifampicin and dexamethasone in the therapy of pneumococcal meningitis; Martinez-Lacasa J et al.; The object of the study was to assess the efficacy of rifampicin and the combination of rifampicin plus vancomycin in a rabbit model of experimental penicillin-resistant pneumococcal meningitis . We also studied the effect of concomitant dexamethasone on the CSF antibiotic levels and inflammatory parameters . The rabbit model of pneumococcal meningitis was used . Groups of eight rabbits were inoculated with 106 cfu/mL of a cephalosporin-resistant pneumococcal strain (MIC of cefotaxime/ceftriaxone 2 mg/L) . Eighteen hours later they were treated with rifampicin 15 mg/kg/day, vancomycin 30 mg/kg/day or both plus minus dexamethasone (0.25 mg/kg/day) for 48 h . Serial CSF samples were withdrawn to carry out bacterial counts, antibiotic concentration and inflammatory parameters . Rifampicin and vancomycin promoted a reduction of >3 log cfu/mL at 6 and 24 h, and cfu were below the level of detection at 48 h . Combination therapy with vancomycin plus rifampicin was not synergic but it had similar efficacy to either antibiotic alone and it was able to reduce bacterial concentration below the level of detection at 48 h . Concomitant use of dexamethasone decreased vancomycin levels when it was used alone (P< 0.05), but not when it was used in combination with rifampicin . Rifampicin alone at 15 mg/kg/day produced a rapid bactericidal effect in this model of penicillin-resistant pneumococcal meningitis . The combination of vancomycin and rifampicin, although not synergic, proved to be equally effective . Using this combination in the clinical setting may allow rifampicin administration without emergence of resistance, and possibly concomitant dexamethasone administration without significant interference with CSF vancomycin levels.

Diagn Microbiol Infect Dis, 2002 Feb, 42(2), 123 - 8
In vitro activity of clinafloxacin in comparison with other quinolones against Stenotrophomonas maltophilia clinical isolates in the presence and absence of reserpine; Ribera A et al.; A total of 33 Stenotrophomonas maltophilia clinical isolates were tested for their susceptibility to clinafloxacin in comparison with ciprofloxacin, levofloxacin, moxifloxacin, nalidixic acid, norfloxacin, sparfloxacin and trovafloxacin . The MIC(50) and MIC(90) were as follows: ciprofloxacin 4 and 64 microg/mL; clinafoxacin 0.5 and 4 microg/mL; levofloxacin 2 and 32 microg/mL; moxifloxacin 1 and 8 microg/mL; nalidixic acid 8 and 128 microg/mL; norfloxacin 64 and 256 microg/mL; sparfloxacin 1 and 16 microg/mL; and trovafloxacin 1 and 8 microg/mL . Clinafloxacin was the most active quinolone, with only a 15.1% of strains showing resistance . When the MICs were determined in the presence of 25 microg/ml of reserpine, the MIC(90) of trovafloxacin and moxifloxacin did not change, whereas decreased 2-fold for clinafloxacin, levofloxacin, sparfloxacin and nalidixic acid, and 4- and 8-fold for ciprofloxacin and norfloxacin respectively . No clinafloxacin-resistant strains were observed when the MIC was performed in the presence of reserpine . Therefore, clinafloxacin shows the better "in vitro"activity against these 33 strains of S.maltophilia.

Yonsei Med J, 2002 Feb, 43(1), 59 - 64
Mutations in the embB locus among Korean clinical isolates of Mycobacterium tuberculosis resistant to ethambutol; Lee HY et al.; Resistance of Mycobacterium tuberculosis to ethambutol (EMB) has been assigned to an operon, embCAB, which has been proposed to be a structural gene for mycobacterial arabinosyl transferases . Recently, genetic events resulting in structural mutations at embB have been proposed as major contributors to the EMB-resistance of isolates whose minimum inhibitory concentration (MIC) level is higher than 20 microgram/ml . On the contrary, isolates with a MIC level lower than 20 microgram/ml do not seem to contain any sequence alterations . In this study, in an effort to understand the role of embB mutations at a low-level of EMB resistance, we investigated the sequence polymorphisms of clinical isolates whose MIC levels are lower than 10 microgram/ml . Accordingly, the sequence alterations of a 312-bp region of the embB gene containing the 306th codon, which has been assigned as a hot-spot for EMB-resistance related mutations, were determined for 21 EMB-resistant and 5 EMB-susceptible clinical isolates . In brief, among 21 EMB- resistant isolates examined, 12 (57.1%) contained mutations in embB (10 at the 306th codon and 2 at other sites), and the remaining isolates 9 contained no mutations in any region of embB . The observed mutations included M306V, M306I, and M306L substitutions that have been reported previously . However, 3 were novel types, which included M306T, A313G and Y319C, D328Y {corrected} double substitutions . On the other hand, all of the EMB-susceptible isolates were found to be free of mutations . In conclusion, our findings suggest that sequence polymorphism of embB may play a pivotal role in the EMB- resistance of M . tuberculosis.

Antimicrob Agents Chemother, 2002 Mar, 46(3), 871 - 4
In vitro activities of BAL9141, a novel broad-spectrum pyrrolidinone cephalosporin, against gram-negative nonfermenters; Zbinden R et al.; The activities of BAL9141 (formerly Ro 63-9141), a novel pyrrolidinone-3-ylidenemethyl cephalosporin, against 244 strains of gram-negative nonfermenters were evaluated . The overall MIC at which 50% of isolates are inhibited (MIC50) and the overall MIC90 were 2 and 64 microg/ml, respectively, which are similar to those of imipenem, lower than those of the other cephalosporins tested, amoxicillin, and the ticarcillin-clavulanic acid combination, and much higher than those of ciprofloxacin . BAL9141 shows species-dependent activity in vitro against a variety of gram-negative nonfermentative pathogens.

Antimicrob Agents Chemother, 2002 Mar, 46(3), 859 - 62
Optimal dose of amoxicillin in treatment of otitis media caused by a penicillin-resistant pneumococcus strain in the gerbil model; Parra A et al.; Amoxicillin at doses of 0.2 to 5 mg/kg of body weight was administered for the treatment of pneumococcal otitis media in a gerbil model . Doses greater than or equal to 2.5 mg/kg, which resulted in concentrations in middle ear fluid of > or = 1.4 microg/ml and concentrations in serum higher than the MIC (1 microg/ml) for > or = 14% of the dosing interval, were both clinically and bacteriologically effective.

Antimicrob Agents Chemother, 2002 Mar, 46(3), 739 - 45
Single- and multiple-dose pharmacokinetics of caspofungin in healthy men; Stone JA et al.; Caspofungin, a glucan synthesis inhibitor, is being developed as a parenteral antifungal agent . The pharmacokinetics of caspofungin following 1-h intravenous infusions in healthy men was investigated in four phase I studies . In an alternating two-panel (six men each), rising-single-dose study, plasma drug concentrations increased proportionally with the dose following infusions of 5 to 100 mg . The beta-phase half-life was 9 to 10 h . The plasma drug clearance rate averaged 10 to 12 ml/min . Renal clearance of unchanged drug was a minor pathway of elimination (approximately 2% of the dose) . Multiple-dose pharmacokinetics were investigated in a 2-week, serial-panel (5 or 6 men per panel) study of doses of 15, 35, and 70 mg administered daily; a 3-week, single-panel (10 men) study of a dose of 70 mg administered daily; and a parallel panel study (8 men) of a dose of 50 mg administered daily with or without a 70-mg loading dose on day 1 . Moderate accumulation was observed with daily dosing . The degree of drug accumulation and the time to steady state were somewhat dose dependent . Accumulation averaged 24% at 15 mg daily and approximately 50% at 50 and 70 mg daily . Mean plasma drug concentrations were maintained above 1.0 microg/ml, a target selected to exceed the MIC at which 90% of the isolates of the most clinically relevant species of Candida were inhibited, throughout therapy with daily treatments of 70 or 50 mg plus the loading dose, while they fell below the target for the first 2 days of a daily treatment of 50 mg without the loading dose . Caspofungin infused intravenously as a single dose or as multiple doses was generally well tolerated . In conclusion, the pharmacokinetics of caspofungin supports the clinical evaluation of once-daily dosing regimens for efficacy against fungal infections.

Antimicrob Agents Chemother, 2002 Mar, 46(3), 702 - 7
Comparison of fractional inhibitory concentration index with response surface modeling for characterization of in vitro interaction of antifungals against itraconazole-susceptible and -resistant Aspergillus fumigatus isolates; Te Dorsthorst DT et al.; Although the fractional inhibitory concentration (FIC) index is most frequently used to define or to describe drug interactions, it has some important disadvantages when used for drugs against filamentous fungi . This includes observer bias in the determination of the MIC and no agreement on the endpoints (MIC-0, MIC-1, or MIC-2 {> or = 95, > or = 75, and > or = 50% growth inhibition, respectively}) when studying drug combinations . Furthermore, statistical analysis and comparisons are troublesome . The use of a spectrophotometric method to determine the effect of drug combinations yields quantitative data and permits the use of model fits to the whole response surface . We applied the response surface model described by Greco et al . (W . R . Greco, G . Bravo, and J . C . Parsons, Pharmacol . Rev . 47:331-385, 1995) to determine the interaction coefficient alpha (ICalpha) using a program developed for that purpose and compared the results with FIC indices . The susceptibilities of amphotericin B (AM), itraconazole (IT), and terbinafine (TB) were tested either alone or in combination against 10 IT-susceptible (IT-S) and 5 IT-resistant (IT-R) clinical strains of Aspergillus fumigatus using a modified checkerboard microdilution method that employs the dye MTT {3-(4,5-dimethyl-2-thiazyl)2,5-diphenyl-2H-tetrazolium bromide} . Growth in each well was determined by a spectrophotometer . FIC indices were determined and ICalpha values were estimated for each organism strain combination, and the latter included error estimates . Depending on the MIC endpoint used, the FIC index ranged from 1.016 to 2.077 for AM-IT, from 0.544 to 1.767 for AM-TB, and from 0.656 to 0.740 for IT-TB for the IT-S strains . For the IT-R strains the FIC index ranged from 0.308 to 1.767 for AM-IT, from 0.512 to 1.646 for AM-TB, and from 0.403 to 0.497 for IT-TB . The results indicate that the degree of interaction is not only determined by the agents themselves but also by the choice of the endpoint . Estimates of the ICalpha values showed more consistent results . Although the absolute FIC indices were difficult to interpret, there was a good correlation with the results obtained using the ICalpha values . The combination of AM with either IT or TB was antagonistic in vitro, whereas the combination of IT and TB was synergistic in vitro for both IT-S and IT-R strains . The use of response surface modeling to determine the interaction of drugs against filamentous fungi is promising, and more consistent results are obtained by this method than by using FIC indices.

Clin Microbiol Infect, 2001 Dec, 7(12), 703 - 5
Activity of telithromycin against 26 quinolone-resistant pneumococci with known quinolone-resistance mechanisms; Nagai K et al.; NCCLS agar dilution was used to test activity of telithromycin compared to clarithromycin, penicillin G, ciprofloxacin, levofloxacin, sparfloxacin and moxifloxacin against 26 pneumococci with defined quinolone resistance (type II topoisomerase and efflux) mechanisms . Thirteen strains were penicillin susceptible, six intermediate and seven resistant . Clarithromycin resistance (mef and/or erm) was seen in eight strains . Ciprofloxacin MICs (mg/L) were 8-64 compared to 1-32 (levofloxacin), 0.5 . or = 32 (sparfloxacin) and 0.125-4 (moxifloxacin) . Telithromycin MIC50 and MIC90 values (mg/L) were 0.016 and 0.25, with only one strain having an MIC of 2 mg/L.

Eur J Immunogenet, 2002 Feb, 29(1), 35 - 46
Further polymorphism of the MICA gene; Perez-Rodriguez M et al.; The MHC class I chain-related (MIC) gene family constitutes an interesting genetic group that is related to major histocompatibility complex (MHC) class I genes and is located within the MHC . The MIC gene products, MICA and MICB, have similar structures to HLA class I molecules . So far over 50 MICA alleles have been reported, which suggests that this genetic system is highly polymorphic . In order to investigate further the extent of MICA polymorphism we have studied exons 2-5 of the MICA gene in over 200 homozygous and heterozygous cell lines . Altogether we have identified 11 new MICA alleles and report 13 new nucleotide variations, one in exon 2, four in exon 3, four in exon 4, two in intron 1, one in intron 4 and one (a deletion) in exon 4 . Eight of the 10 exonic variations are non-synonymous . The deletion in exon 4 leads to a frame-shift mutation and the introduction of a repeat of 12 leucine residues encoded by the microsatellite in exon 5 . This study provides further evidence that the MICA gene is highly polymorphic . In contrast to MHC class I molecules, the polymorphic sites in MICA are predominantly within the alpha2 and alpha3 domains . The distribution of synonymous and non-synonymous substitutions suggests that there is selection for the polymorphic positions, which therefore define potential functional sites in the protein . We were also able to determine the association between MICA and HLA-B alleles in a number of homozygous cell lines bearing extended haplotypes.

Pediatr Pulmonol, 2002 Mar, 33(3), 167 - 73
Determinants of bronchial responsiveness to methacholine at school age in twin pairs; Nikolajev K et al.; The methacholine inhalation challenge test (MIC) was used to evaluate bronchial responsiveness in 67 children who were the products of multiple pregnancies when they were 7-15 years old . At birth, 30 (45%) infants had intrauterine growth retardation (IUGR; birth weight <2 SD below normal birth weight, or birth weight difference >1.3 SD between twin-pairs), and 59 (88%) were born before 37 weeks of gestation . None of the children had doctor-diagnosed asthma . The provocative dose of methacholine causing a 20% fall in Wright's peak expiratory flow (WPEF) (PD20) was below 1,000 microg in 10 (15%) children, and they were classified as MIC responders . There were no differences in perinatal or neonatal factors between MIC responders and nonresponders; in particular, MIC responses did not differ between IUGR infants, and children with appropriate growth for gestational age (AGA) at birth . There were seven discordant pairs in which one child was a MIC responder and the other was not; 5 responders were IUGR, and 2 were AGA children (ns) . Respiratory tract infections after the neonatal period were equally common in IUGR and AGA children . However, these infections were associated with later bronchial hyperresponsiveness . Doctor-diagnosed respiratory infections, numbers of antibiotic courses, episodes of otitis media, and the need for adenoidectomy, tonsillectomy, and tympanostomy were more common in MIC responders than in nonresponders . We conclude that IUGR was not associated with subsequent bronchial hyperresponsiveness in twin pairs assessed by the MIC test . A significant relationship was seen between bronchial hyperresponsiveness and infections after the neonatal period .

Zhonghua Wai Ke Za Zhi, 2000 Feb, 38(2), 137 - 9
{Experimental and clinical study of gentamicin-loaded chitosan drug delivery system}; Chen A et al.; OBJECTIVE: To conduct a clinical study and release test for a drug delivery system (DDS) of gentamicin-loaded chitosan bar in attempt to extend its clinical application . METHODS: The diffusion behavior of gentamicin from chitosan DDS was detected using in vitro, in vivo release test . Eighteen cases of chronic osteomyelitis were treated by chitosan DDS . The clinical result was evaluated by wound healing and clinical and X-radiographic manifestation . RESULTS: Concentration of gentamicin about 926.7 microg/bar/day was released from DDS within the first 24 hours by in vitro diffusion test . It fell and sustained by 25th day with rate of 4.0 microg x bar(-1) x day(-1) . In in vivo study, serum gentamicin concentration reached its peak level (0.92 microg/ml) 24 hours after implantation . No increase was observed in the concentration of BUN and Cr . In all bone tissue around the bar, 8 weeks after oderation the gentamicin concentration exceeded the minimum inhibitory concentration for the common causative organisms of osteomyelitis . The follow up time was 24.8 months (6 - 34) . The initial cure ratio was 2/18 and the recurrence ratio zero . CONCLUSION: The gentamicin-loaded chitosan bar is a good DDS with sustained antibiotic effect in vivo and in vitro . It is an effective method for the treatment of bone infection.

J Antibiot (Tokyo), 2001 Nov, 54(11), 967 - 72
Mutagenesis of the rapamycin producer Streptomyces hygroscopicus FC904; Cheng YR et al.; Rapamycin (RPM) is produced by Streptomyces hygroscopicus FC904 isolated from soil in Fuzhou, China . It is a triene macrolide antibiotic with potential application as an immunosuppressant and drug for human gene therapy . In an attempt to improve rapamycin production, mutation and screening of the parent culture have been carried out . Thousands of survivors were obtained after mutagenesis by NTG (3 mg/ml) and UV (30 W, 15 cm, 30 seconds) of spore suspensions . None showed improved production of RPM . We determined the susceptibility to antibiotics of S . hygroscopicus FC904 by two fold dilutions of antibiotics in oatmeal agar plates . It was found that the strain was resistant to penicillin, erythromycin, RPM, tetracycline and chloramphenicol, but susceptible to mitomycin C (MIC, 10 microg/ml) and aminoglycosides such as gentamicin (MIC, 0.1 microg/ml), kanamycin (MIC, 0.1 microg/ml) and streptomycin (MIC, 0.3 microg/ml) . Protoplasts of strain FC904 were prepared after finding the best conditions for their formation . They were treated with gentamicin, erythromycin, mitomycin C and NTG . Surprisingly, gentamicin was especially effective for obtaining higher RPM-producing mutants . Mutant C14 was selected by exposing the protoplasts of the parent strain FC904 to 1 microg/ml of gentamicin at 28 degrees C for 2 hours . A higher RPM-producing mutant (C14-1) was obtained from the protoplasts of mutant C14 treated with gentamicin, and its titer was 60% higher than that of the parent strain FC904 by HPLC analysis . Another improved mutant (C14-2) was obtained from the spores of mutant C 14 treated with 1 microg/ml of gentamicin plus 2 mg/ml of NTG at 28 degrees C for 2 hours . Mutant C14-2 had a titer 124% higher than FC904 . The possible mechanism for the effect of gentamicin by using protoplasts or spore suspensions will be discussed, i.e . the possibility of gentamicin being a mutagen or a selective agent.

Transplantation, 2002 Jan 27, 73(2), 304 - 6
MIC expression in renal and pancreatic allografts; Hankey KG et al.; BACKGROUND: MHC class I chain-related antigen A (MICA) and MHC class I chain-related antigen B (MICB) are HLA class I related products of polymorphic MHC genes . Constitutive expression in normal tissue is limited to gut epithelium but can be induced in other epithelial cells by stress . Specific antibodies against MICA have been reported in the serum of patients who had rejected kidney allografts, suggesting a potential role for these molecules in transplant immunopathology . However, expression of MICA and MICB in transplanted organs has not been demonstrated . In this study, we report the expression of MICA and MICB in renal and pancreatic allograft biopsies, which were obtained due to clinical signs of rejection . METHODS: A monoclonal antibody directed against MICA and MICB was used to perform indirect immunohistochemistry on formalin fixed, paraffin embedded needle biopsies of kidney and pancreas allografts . The results of staining were then compared to the standard light microscopic evaluation of the biopsies for rejection . RESULTS: A total of 53 individual renal transplant biopsies and 19 pancreas transplant biopsies were assayed for expression of MIC . Histologically, renal biopsies were diagnosed as no rejection, acute tubular necrosis (ATN), acute rejection (AR), chronic rejection (CR), and acute and chronic rejection (ACR) . No staining was observed in 7 of 10 kidneys showing no rejection . All 11 of the kidney biopsies with AR were positive, as were the 11 ATN cases, 9 of the 11 kidney biopsies with CR, and 7 of the 10 with ACR . The acini of normal, nontransplanted, pancreas, control specimen were consistently negative; however, islets were positive in all specimens . The acini and islets of five histologically normal pancreas biopsies were positive, as were the four biopsies with AR, seven biopsies with CR, and two with ACR . CONCLUSIONS: MICA and MICB are expressed in epithelial cells in allografted kidney and pancreas that show histologic evidence of rejection and/or cellular injury . In addition to previous findings of alloantibodies against MICA, expression of these gene products may play a role in allograft rejection.

World J Gastroenterol, 1998 Dec, 4(6), 536 - 539
Preliminary study on the production of transgenic mice harboring hepatitis B virus X gene; Zhu HZ et al.; AIM:To establish transgenic mice lineage harboring hepatitis B virus X gene and to provide an efficient animal model for studying the exact role of the HBx gene in the process of hepatocarcinogenesis.METHODS:The HBx transgenic mice were produced by microinjecting the construct with X gene of HBV (subtype adr) DNA fragment into fertilzed eggs derived from inbred C57BL/6 strain; transgenic mice were identified by using Nested PCR; expression and phenotype of HBx gene were analyzed in liver from transgenic mic at the age of 8 weeks by RT-PCR, pathologic examination and periodic acid-schiff staining (PAS), respectively.RESULTS:Five hundred and fourteen fertilized eggs of C57 BL/6 mice were microinjected with recombinant retroviral DNA fragment, and 368 survival eggs injected were transferred to the oviducts of 18 pseudopregnant recipient mice, 8 of them became pregnant and gave birth to 20 F1 offspring . Of 20 offsprings, four males and two females carried the hybrid gene (HBx gene) . Four male mice were determined as founder, named X 1, X 5, X 9 and X 15 . These founders were back crossed to set up F1 generations with other ibred C57BL/6 mice or transgenic littermates, respectively.Transmission of HBx gene in F1 offspring of X 1, X 5 and X 9 except in X 15 followed Mendelian rules . The expression of HBx mRNA was detected in liver of F1 offspring from the founder mice (X 1 and X 9), which showed vacuolation lesion and glycogen positive foci.CONCLUSION:Transgenic mice harboring HBx gene were preliminarily established.

Pharmazie, 2001 Nov, 56(11), 871 - 4
Antimycobacterial activity of 4'-bromo-{1,1'-biphenyl}-4-yl 4-x-phenylmethanone derivatives, and their acute toxicity and cytotoxicity; De Souza AO et al.; The antimycobacterial activity of nine biphenyl methanone (BPM) derivatives against standard strains of Mycobacterium kansasii, M . avium and M . malmoense was determined by colorimetric assay in microplates with the dye Alamar Blue . Acute toxicity of these compounds was also analyzed by determination of CO2 concentration in a respirometric assay using Escherichia coli . The compounds showed weak antimycobacterial activity with a minimal inhibitory concentration (MIC) over 0.038 mmol l-1 and no toxicity was found in E . coli up to 400 mmol l-1 . No cytotoxicity was observed on V79 cells up to 0.35 mmol l-1 with 7 of the BPM derivatives, with two exceptions (X = SO2CH3, NO2) that showed some toxicity . The greatest antimycobacterial activity was observed with the SO2CH3 derivative and the application of Principal Component Analysis (PCA) showed a relationship between structure and antimycobacterial activity of the compounds . Two descriptors, nucleophilic superdelocalizability of carbon atom and pi-hydrophobic constant, were necessary to describe this relationship.

Ann Pharmacother, 2002 Jan, 36(1), 63 - 6
Severe cellulitis/myositis caused by Stenotrophomonas maltophilia; Downhour NP et al.; OBJECTIVE: To present a case of cellulitis/myositis due to Stenotrophomonas maltophilia in the absence of trauma and to discuss a potentially novel treatment option . CASE SUMMARY: A 57-year-old white man, having undergone an allogeneic bone marrow transplant, developed myositis with S . maltophilia of the left soleus muscle; there had been no trauma . Risk factors for infection included neutropenia, prolonged hospitalization and intensive care unit stay, and broad-spectrum antibiotic exposure . The affected area of muscle was resected and the patient successfully treated with trimethoprim/sulfamethoxazole (TMP/SMX), ticarcillin/clavulanate, and aztreonam . DISCUSSION: In severe myositis/cellulitis caused by S . maltophilia, TMP/SMX is considered the drug of choice . However, bacteriostatic agents such as TMP/SMX are less than ideal in neutropenic patients . The combination of ticarcillin/clavulanate plus aztreonam has been shown to improve activity in vitro against this organism compared with TMP/SMX . This is likely due to inhibition of the 2 beta-lactamases this organism produces by clavulanate and aztreonam . In our study of clinical isolates of S . maltophilia, this combination reduced the minimum inhibitory concentration at 90% by 128-fold and was synergistic against 10 of 12 isolates tested in time-kill analysis . CONCLUSIONS: S . maltophilia is emerging as an important pathogen in patients with compromised immunity, leading to severe infections that are difficult to treat . Based on in vitro synergy studied, we recommend considering ticarcillin/clavulanate plus aztreonam as a potential treatment option in immunocompromised patients with S . maltophilia infection.

Pediatr Dev Pathol, 2002 Jan-Feb, 5(1), 45 - 53
Cutaneous lymphoblastic lymphoma in children: report of six cases with precursor B-cell lineage; Kahwash SB et al.; Precursor B lymphoblastic lymphomas (B-LBL) are generally rare, but appear to have a higher incidence in children than in adults . In this report, we describe in detail six cases of B-LBL presenting with cutaneous lesions . Three occurred in the scalp, one in the skin of the thigh, one in the skin of the face and breast, and one in the subcutaneous tissue of the orbit . All six patients are females ranging in age at presentation from 5 to 15 years (mean = 9.6) . None of the cases had bone marrow involvement, while two had bone involvement (maxilla, distal tibia, and distal humerus in one case, and distal tibia and orbital bone in another case); only one case had lymphadenopathy (retroperitoneal) . Immunohistochemical staining showed positivity for CD79a and CD43 in all six cases . LCA and L26 positivity were also each seen in one case . Staining for MIC-2 (CD99) showed strong positivity in three cases . Vimentin was positive in four cases and TdT was positive in all five patients tested . Staining for keratin, UCHL-1, or CD30 was not encountered . Cases in which cell marker studies by flow cytometry were performed showed positivity for CD10, CD19 with negative CD20, pan-T-cell, and myeloid markers . The five patients who received multiagent chemotherapy are alive with follow-up intervals of 2 to 18 years . Two patients had local recurrences and were given radiation therapy (one with repeating multiagent chemotherapy) . One patient (diagnosed in 1962) died of disseminated disease; she had been treated with radiation therapy and 6MP only . Cutaneous B-LBL must be included in the differential diagnosis of small blue cell tumors, especially in children . In contrast to its T-cell counterpart, B-LBL occurs more frequently in females, tends to present as skin or bone lesions, and is associated with a potential cure, even in cases that relapse.

J Antimicrob Chemother, 2002 Feb, 49(2), 327 - 30
Cefepime is efficacious against penicillin- and quinolone-resistant pneumococci in experimental meningitis; Cottagnoud P et al.; In experimental rabbit meningitis, cefepime given at a dose of 100 mg/kg was associated with concentrations in the cerebrospinal fluid of between 5.3 and 10 mg/L and a bactericidal activity of -0.61 +/- 0.24 Delta log(10) cfu/mL x h, similar to the standard regimen of ceftriaxone combined with vancomycin (-0.58 +/- 0.14 Delta log(10) cfu/mL x h) in the treatment of meningitis due to a penicillin- and quinolone-resistant pneumococcal mutant strain (MIC 4 mg/L) . Compared with the penicillin-resistant parental strain, the penicillin- and quinolone-resistant mutant was killed more slowly by cefepime and ceftriaxone in time-killing assays in vitro over 8 h.

Int J Antimicrob Agents, 2002 Jan, 19(1), 79 - 82
Effect of subminimal inhibitory concentrations of three fluoroquinolones on adherence of uropathogenic strains of Escherichia coli; Baskin H et al.; The effect of subinhibitory concentrations (1/2-1/32 x MIC) of ciprofloxacin, ofloxacin and levofloxacin on the adherence of three strains of Escherichia coli (a mannose-resistant haemagglutinating clinical isolate, a non-haemagglutinating clinical isolate and the mannose-resistant haemagglutinating ATCC 25922 strain) were studied . Ciprofloxacin had the lowest MIC values but only the 1/2 MIC concentration inhibited adherence of mannose-resistant haemagglutinating strains after exposure to subMIC values . Significant inhibition of adherence was observed with 1/4 x MIC ofloxacin for both haemagglutinating isolate (27096) and the ATCC strain . Levofloxacin might be more effective and safer than ciprofloxacin and ofloxacin as a long acting fluoroquinolone at subMIC values in patients with UTI.

Int J Antimicrob Agents, 2002 Jan, 19(1), 67 - 70
Drug combinations with amoxycillin reduce selection of clarithromycin resistance during Helicobacter pylori eradication therapy; Murakami K et al.; This study investigated the relationship between the drug combinations of the eradication regimens and the prevalence of acquired resistance to clarithromycin . Of 540 patients treated with anti-Helicobacter pylori regimens containing clarithromycin, 55 patients (31 males, mean age, 45.6 years) with failed eradication of H . pylori that was susceptible to clarithromycin before treatment were included . The E test was used to test for susceptibility to clarithromycin (minimum inhibitory concentration (MIC) <1 mg/l) . Of the 55 patients, 33 (60.0%) developed clarithromycin resistance after failed eradication . Of the dual therapies, the combination of a proton pump inhibitor (PPI) and clarithromycin resulted in 88.9% (8/9) of the patients acquiring clarithromycic-resistance . With the triple therapies, the percentages of patients acquiring clarithromycin resistant strains after using a PPI+clarithromycin+amoxycillin or a PPI+clarithromycin+metronidazole were 38.7% (12/31) and 90.0% (9/10), respectively (P<0.01) . These data suggest that regimens containing amoxycillin may prevent the selection of secondary clarithromycin resistance.

J Infect Chemother, 2000 Jun, 6(2), 101 - 3
Antifungal activity of a new triazole, voriconazole (UK-109496), against clinical isolates of Aspergillus spp; Maesaki S et al.; Voriconazole is a new triazole antifungal agent with potent activity against yeast and molds . We investigated the in-vitro activity of voriconazole compared with that of other antifungal agents against 50 clinical isolates of Aspergillus spp., measured by the National Committee for Clinical Laboratory Standards (NCCLS) reference method described in the M27-A document, and by an alamar blue colorimetric method . Voriconazole was the most potent agent against Aspergillus fumigatus (minimum inhibitory concentration {MIC}90, 0.5 mg/l) and Aspergillus niger (MIC90, 1.0 mg/l) . Voriconazole was less active (MIC90, 1.0 mg/l) against Aspergillus flavus than itraconazole (MIC90, 0.5 mg/l) . Voriconazole was more active than itraconazole against Aspergillus fumigatus and Aspergillus flavus by the alamar blue indicator method for the measurement of MIC . Based on these results, voriconazole has promising activity against commonly encountered isolates of Aspergillus spp., and its clinical usefulness should be established by further studies.

Cornea, 2002 Jan, 21(1), 48 - 50
Residence time of netilmicin in tears; Scuderi AC et al.; PURPOSE: To determine the concentration of netilmicin in tears after eye drop administration . METHODS: A clinical study was carried out on 32 healthy volunteers . Subjects were divided into four groups and underwent tear collection at 5, 10, 20, and 60-minutes after drug administration, respectively . Tear samples were collected through capillary suction from the inferior conjunctival "cul-de-sac" and analyzed by high-pressure liquid chromatography (HPLC) . RESULTS: Netilmicin concentration in tears decreased after a first order kinetics, a best-fit curve was drawn and the minimum inhibitory concentration (MIC90) intersection for the most common ocular pathogens was calculated . CONCLUSIONS: The extrapolation of the curve shows that the concentration of netilmicin on the ocular surface can be effective against microorganisms more than 120 minutes after eye drop instillation.

Vaccine, 2002 Jan 15, 20(7-8), 1106 - 12
Outbreak of aseptic meningitis and mumps after mass vaccination with MMR vaccine using the Leningrad-Zagreb mumps strain; da Cunha SS et al.; Data from routine surveillance during two mass immunisation campaigns (MIC) with Measles-mumps-rubella (MMR)vaccine using Leningrad-Zagreb mumps strain in two states in Brazil were analysed to estimate the risk of vaccine-related meningitis and mumps . Increase in the incidence of the two diseases was observed in both states, 3 weeks after the vaccination campaigns . The estimated number of doses applied per one case of vaccine-related meningitis ranged from 6199 (95% CI: 4854-8058) to 19,247 (95% CI: 12,648-29,513) depending on the diagnostic criteria used and state . It was 300 doses (95% CI: 286-317) for each case of mumps . The implications for vaccination policy are discussed.

Ann Rheum Dis, 2002 Feb, 61(2), 157 - 60
Association of the MIC-A gene and HLA-B51 with Behçet's disease in Arabs and non-Ashkenazi Jews in Israel; Cohen R et al.; BACKGROUND: Behcet's disease is known to be strongly associated with HLA-B51 in many different ethnic groups . Recently, it was suggested that MIC-A (major histocompatibility complex class I related gene A) is the pathogenic gene after strong association was found between the MIC-A A6 allele of the transmembrane region and the disease in Japanese and Greek patients, although in Greek patients this association was found to be due to linkage disequilibrium with HLA-B51 . OBJECTIVES: To investigate microsatellite polymorphism in Arab and non-Ashkenazi Jewish (NAJ) patients in Israel, to determine whether this association occurs in these groups with Behcet's disease, and elucidate the associated HLA allele of the disease . METHODS: Forty four Israeli patients with Behcet's disease, including 20 Arabs and 24 NAJ, and 130 ethnically matched healthy controls were examined for MIC-A microsatellite polymorphism of the transmembrane region using polymerase chain reaction, autoradiography, and sequence analysis . RESULTS: The MIC-A A6 allele was significantly more frequent in the Arab patient group (19/20 (95%)) than in healthy Arab controls (25/42 (60%)) (p(corr)=0.015, OR=12.92), but not in the NAJ patients (16/24 (67%)) compared with NAJ healthy controls (48 /88 (55%)) (p(corr)=1.02, OR=1.667) . In stratification analysis of the Arab subgroup, on the confounding effect of MIC-A A6 on HLA-B51 association and vice versa, Behcet's disease was distinctly associated only with HLA-B51 . CONCLUSIONS: These results imply strong association between the MIC-A A6 allele and the disease in Israeli Arabs, but not in Israeli NAJ patients . The stratification analysis indicates that this association results secondarily from a strong linkage disequilibrium with HLA-B51, and the real disease susceptibility gene which plays a part in the development of Behcet's disease is most probably the HLA-B51 allele itself.

Antimicrob Agents Chemother, 2002 Feb, 46(2), 556 - 7
Azole cross-resistance in Aspergillus fumigatus; Mosquera J et al.; We susceptibility tested 17 clinical isolates of Aspergillus fumigatus, for most of which MICs of itraconazole were elevated (MIC at which 50% of the isolates tested are inhibited, 16 microg/ml), against itraconazole, posaconazole, ravuconazole, and voriconazole . Posaconazole was the most active against itraconazole-susceptible isolates . A complex pattern of cross-resistance and hypersusceptibility was seen with voriconazole and ravuconazole, suggesting marked differences in activity and mechanisms of resistance.

Antimicrob Agents Chemother, 2002 Feb, 46(2), 531 - 3
Sterol and fatty acid composition of Candida lusitaniae clinical isolates; Peyron F et al.; The sterol and fatty acid compositions of four amphotericin B-resistant and of two amphotericin B-susceptible Candida lusitaniae clinical isolates were determined . A flow cytofluorometric susceptibility test (FCST) with a membrane potential-sensitive cationic dye was used as a complement to the conventional method for selecting the isolates . Compared to susceptible isolates, resistant ones showed a greatly reduced ergosterol content and changes in sterol composition, consistent with a defect in Delta8-->7 isomerase . Within each group, no correlation between the sterol or fatty acid pattern or composition and both the degree of in vitro susceptibility and FCST MIC was found.

Antimicrob Agents Chemother, 2002 Feb, 46(2), 517 - 8
In vitro activities of BMS-284756 against Chlamydia trachomatis and recent clinical isolates of Chlamydia pneumoniae; Malay S et al.; The in vitro activities of BMS-284756 (a novel des-fluoroquinolone), levofloxacin, moxifloxacin, and clarithromycin were tested against 5 strains of Chlamydia trachomatis and 20 isolates of Chlamydia pneumoniae . The MIC at which 90% of the isolates were inhibited and the minimal bactericidal concentration at which 90% of the isolates were killed by BMS-284756 for all isolates of C . pneumoniae and C . trachomatis was 0.015 microg/ml (range, 0.015 to 0.03 microg/ml) . BMS-284756 was the most active quinolone tested.

Antimicrob Agents Chemother, 2002 Feb, 46(2), 294 - 9
In vitro antimycobacterial activities of 2'-monosubstituted isonicotinohydrazides and their cyanoborane adducts; Maccari R et al.; As a result of our search for new isoniazid derivatives with extended spectra of activity, we evaluated the in vitro antimycobacterial activities of isonicotinohydrazides (compounds 2) and their cyanoborane adducts (compounds 3), both obtained by the reaction of isonicotinoylhydrazones (compounds 1) with sodium cyanoborohydride . Most of the tested compounds displayed moderate to high activity against Mycobacterium tuberculosis H37Rv, with MICs ranging from 0.2 to 12.5 microg/ml . In particular, some hydrazides showed activity similar to that of rifampin (MIC = 0.2 microg/ml) and rather low cytotoxicity, so that they were generally shown to possess high safety indices . In contrast, the coordination to a cyanoborane (BH(2)CN) group (compounds 3) in general brought about a decrease in antimycobacterial activity, while cytotoxicity increased . Interestingly, selected compounds 1 to 3, mostly hydrazides (compounds 2), were effective in killing M . tuberculosis growing within macrophages at concentrations in culture medium which were much lower than the corresponding MICs . These compounds also displayed good activity against drug-resistant M . tuberculosis strains.

Zhonghua Jie He He Hu Xi Za Zhi, 2000 Feb, 23(2), 89 - 92
{Drug susceptibility testing for M . tuberculosis with proportion method}; Liu Y et al.; OBJECTIVE: To analyze the feasibility of proportion method in drug susceptibility testing (DST) of M . tuberculosis(MTB) . METHODS: DST was performed to 360 strains of MTB using proportion method and absolute concentration method, 30 of them using both China made and Sigma produced anti-TB drugs . MIC was detected in 19 strains whose results were different from standard ones . RESULTS: In isoniazid (INH) and ethambutol (EMB), the resistance rate and the accordance rate compared with standard results were significantly higher in proportion method than absolute concentration method (P < 0.01) . But in streptomycin (SM) and rifampicin (RFP), they did not show any significant difference . The accordance rates of INH, SM, EMB and RFP between two methods were 81.1%, 92.5%, 83.6% and 98.6% . The DST results using China made and Sigma produced anti-TB drugs had no significant difference . CONCLUSIONS: The main reason made proportion method differ from absolute concentration method is critical concentration of drugs . China made anti-TB drugs are also qualified for DST purpose.

Zhonghua Jie He He Hu Xi Za Zhi, 2000 Jan, 23(1), 50 - 4
{Antituberculosis effect of levofloxacin}; Lu Y et al.; OBJECTIVE: Assess the activities of levofloxacin (LVLX) against Mycobacterium tuberculosis both in vitro and in vivo, the pharmacokinetics of LVLX and the effectiveness and safety of it in the treatment of pulmonary tuberculosis, with ofloxacin(OFLX) as control . METHODS: The MIC and MBC of LVLX were determined by the tube doubling dilution method, and the effectiveness of the drugs were assessed by half survival time of the mice . The concentrations of LVLX and OFLX in serum were measured by HPLC . The pharmacokinetic parameters obtained were calculated . 138 newly diagnosed and retreatment pulmonary tuberculosis patients were randomly allocated to receive four chemotherapy regimens . RESULTS: The MIC and MBC of LVLX against Mycobacterium tuberculosis were 2 dilutions lower than that of OFLX, in a murine tuberculosis model, the antimycobacterial activity of LVLX was significant and superior to OFLX with the same dose . The concentration-time curves of LVLX and OFLX were conformed to an one-compartment model . There was no significant difference between the two formulations in the Tmax, T1/2 and 1/2 Cmax, 1/2 AUC of OFLX and LVLX . The sputum conversion rates at the end of treatments of groups I, II, III and IV were 97%, 97%, 82% and 84% respectively . X-ray resolution rates were 97%, 94%, 88% and 88% respectively . The side effects of four groups were rather low . CONCLUSIONS: LVLX displays powerful activities against Mycobacterium tuberculosis both in vitro and in vivo, which is two times that of OFLX.LVLX and OFLX have the same good pharmacokinetic characteristics . LVLX at the dose of 300 mg/d shows the same effectiveness and fewer adverse drug reactions in comparison with OFLX at the dose of 600 mg/d in the treatment of pulmonary tuberculosis . So LVLX is a new effective and safe antituberculosis drug.

Diagn Microbiol Infect Dis, 2001 Dec, 41(4), 211 - 4
Aspergillus: rising frequency of clinical isolation and continued susceptibility to antifungal agents, 1994-1999; Chandrasekar PH et al.; We investigated the frequency of clinical isolation and the in vitro susceptibility to antifungal agents of Aspergillus species obtained from patients at the Detroit Medical Center from January 1994 to December 1999 . During this period, 593 clinical isolates of Aspergillus species {406 A . fumigatus, 68%; 82 A . niger, 14%; 42 A . flavus, 7%; 63 Aspergillus spp., 11%} were recovered from hospitalized patients . From January 1996 to December 1999, approximately 2.5-4.5 fold yearly increase of the number of aspergillus isolates occurred compared to that of 1994 . Conidial suspensions from clinical isolates were prepared and their in vitro susceptibility to amphotericin B and three azoles were determined . All four agents examined were extremely active . The minimum inhibitory concentrations (MIC(90)) (microg/mL) of amphotericin B, itraconazole, voriconazole and posaconazole for A . fumigatus (n = 406) were 0.5, 1.0, 0.5 and 0.25 . Similar values were noted for non-A . fumigatus isolates . A year-to-year comparison of the MIC(90) of the four agents for A . fumigatus and non-A . fumigatus isolates over the 6-year study period showed no significant differences . Our study showed a steady increase in the frequency of clinical isolation of Aspergillus species; and the organism has remained susceptible to amphotericin B/triazoles without any change in susceptibility levels during the 6-year study period.

Anal Quant Cytol Histol, 2001 Dec, 23(6), 405 - 12
Flow cytometric immunophenotyping and comparison with immunocytochemistry in small round cell tumors; Brahmi U et al.; OBJECTIVE: To quantitate different antigens by flow cytometric immunophenotyping (FCI) in small round cell tumors (SRCTs) and to compare the FCI technique with immunocytochemistry (IC) . STUDY DESIGN: IC and FCI were performed on 24 consecutive cases of SRCT on fine needle aspiration biopsy material using a panel of antibodies--e.g., cytokeratin (CK), leukocyte common antigen (LCA), desmin, epithelial membrane antigen, neuron-specific enolase, chromogranin, retinoblastoma gene product, neuroblastoma clone (NB84a (NB), vimentin and Mic-2 gene product . IC was done by indirect immunoperoxidase and FCI by indirect immunofluorescence onflow cytometry . RESULTS: In Ewing's sarcoma, with the help of FCI, positive results were obtained in an additional 4 samples in CK, 2 samples in actin and 3 samples in desmin . Similarly, one each sample was additional positive regarding Mic-2 and vimentin by IC . In cases of neuroblastoma with the help of FCI, additional positive results were obtained in one each sample of CK, LCA and NB and two in actin . Combined use of FCI and IC helped to show chromogranin positivity in an additional two cases . Divergent differentiation was noted in four cases of Ewing's sarcoma, one neuroblastoma and two peripheral neuroectodermal tumors . CONCLUSION: FCI technique is sensitive, more objective and quantitative in comparison with manual absorbance-based microscopic detection of enzyme immunohistochemistry products . FCI may determine divergent differentiation in SRCTs.

J Ocul Pharmacol Ther, 2001 Dec, 17(6), 555 - 63
Ocular bioavailability of ciprofloxacin in sustained release formulations; Ke TL et al.; A novel sustained release delivery system of ciprofloxacin for the eye was developed . The system consists of a viscosity enhancer (carbopol gel or hydroxypropylmethylcellulose solution) plus a penetration enhancer (dodecylmaltoside) to overcome penetration barriers and loss due to wash-out and thus achieve the desired ciprofloxacin ocular absorption . The present studies were designed to assess the ocular penetration and bioavailability of ciprofloxacin in sustained release formulations . In vitro studies in rabbits indicated an approximate 10-fold increase in drug penetration through the rabbit cornea using the penetration enhancer, dodecylmaltoside . In vivo bioavailability studies demonstrate that these formulations provided a long drug duration in the cornea . After administration of a single topical dose of ciprofloxacin (0.3%/30 microL), corneal levels greater than the Minimum Inhibitory Concentration (MIC90) (0.5 microg/g) were observed through eight hours . These sustained release formulations delivered 10-fold more drug into the aqueous humor than the standard solution formulation . Maximum ciprofloxacin concentrations in the aqueous humor (0.5-0.7 microg/mL) were attained between one and two hours after dosing . Using these sustained release formulations, ciprofloxacin can penetrate to the anterior chamber of the eye in concentrations that are inhibitory for most gram-negative and gram-positive organisms . These topical ocular formulations have prophylactic utility for prevention of post-surgical infection, offering greater efficacy and safety than currently available treatments.

Zhonghua Jie He He Hu Xi Za Zhi, 1999 Jun, 22(6), 354 - 7
{A series of clinical study on netilmicin}; Ji S et al.; OBJECTIVE: To evaluate the clinical effects, pharmacokinetics, post-antibiotic effect (PAE) and toxicity of netilmicin as a single daily dose in the treatment of lower respiratory tract infection . METHODS: 48 cases were divided into 3 groups: In the first group, Netilmicin(6 mg.kg-1.d-1) was administered in a single daily dose; in the second group, netilmicin (200 mg/d) was combined with cefazolin (3 g, Q12 h); and in the third control group, the combination of cefazolin and Amikacin was used . Pharmacokinetics were studied in 7 patients using the TDX system, and PAE induced by Netilmicin was determined by the Avantage microbiologic system . Clinical symptoms, laboratory studies, chest X-rays, and side effects were observed . RESULTS: The overall clinical effects of the first group were better than those of the third group . The mean serum concentration of netilmicin was 27.23 mg/L, the valley serum concentration was 0.23 mg/L, T1/2 beta was 5.059 h, AUC was 70 micrograms.h-1.ml-1.netilmicin at concentrations 0.5, 1.0 and 4 times the MIC showed different degrees of PAE against 4 strains of bacterium . Nephrotoxicity and ototoxicity were not found in the treatment group . CONCLUSIONS: Netilmicin in a single daily dose resulted in a high peak serum concentration and big AUC . As a concentration-dependent bactericidal agent, netilmicin showed a longer PAE and better therapeutic effects.

Rev Esp Quimioter, 2001 Sep, 14(3), 281 - 5
{Activity of itraconazole against clinical isolates of Aspergillus spp . and Fusarium spp . determined by the M38-P NCCLS method}; Carrillo-Munoz AJ et al.; The antifungal activity of itraconazole was studied in 101 clinical isolates of Aspergillus fumigatus, A . flavus, A . niger, A . terreus, A . nidulans, A . candidus, A . glaucus, A . clavatus, Fusarium solani, F . oxysporum and F . semitectum . The minimum inhibitory concentrations (MIC) were determined according to the protocol of the M38-P National Committee for Laboratory Standards (NCCLS) document using a microdilution method in 1640 RPMI liquid medium (visual reading at 48 and 72 h incubation) . In general, the MIC did not vary with time of incubation, except in a Z . fumigatus strain in which the MIC went from 2 to 16 mg/l . The geometric mean of the MIC and MIC(90) of itraconazole for Aspergillus spp . was 0.44 mg/l and 0.5 mg/l, respectively; and for Fusarium spp . it was 14.1 mg/l and 16 mg/l, respectively . With 0.5 mg/l 75% of the Aspergillus spp . strains were inhibited, and 100% of these strains were inhibited with 2 mg/l . A . niger and A . fumigatus were the most resistant species (MIC(90) 2 mg/l) . The MIC of all the Fusarium strains essayed was between 4 and 16 mg/l.

J Antimicrob Chemother, 2002 Jan, 49(1), 209 - 13
Resistance to amphotericin B does not emerge during treatment for invasive aspergillosis; Moosa MY et al.; Emergence of resistance to antifungal drugs during therapy for invasive aspergillosis has received scant attention . We recovered Aspergillus isolates from six patients with invasive aspergillosis, who were receiving amphotericin B before fungal isolation . Although isolates were susceptible to amphotericin B in vitro, none of the patients survived . The MIC of amphotericin B for isolates was similar to that for isolates from 35 patients with no prior exposure to amphotericin B . Laboratory attempts to produce amphotericin B resistance in Aspergillus were unsuccessful . These data indicate that emergence of resistance to amphotericin B is uncommon during therapy for invasive aspergillosis.

J Antimicrob Chemother, 2002 Jan, 49(1), 129 - 34
Clearance of ceftazidime during continuous venovenous haemofiltration in critically ill patients; Traunmuller F et al.; Published recommendations for the optimal dosing regimen of ceftazidime in critically ill patients with continuous venovenous haemofiltration (CVVH) differ . The aim of this prospective study was to analyse the pharmacokinetic and pharmacodynamic parameters of ceftazidime during CVVH with a high-flux polysulphone membrane, and derive a dosage recommendation . Twelve critically ill patients (five female, seven male) with acute renal failure undergoing CVVH using a 0.7 m(2) polysulphone high-flux membrane were investigated . All patients received ceftazidime 2 g i.v . q8h . Peak ceftazidime concentrations were 58.2 +/- 11.6 mg/L, with trough concentrations 14.0 +/- 3.2 mg/L at the arterial port . The elimination half-life, haemofiltration clearance, volume of distribution and total removal were 4.3 +/- 0.6 h, 32.1 +/- 7.9 mL/min, 36.4 +/- 6.4 L and 74.5 +/- 6.5%, respectively . Based on these pharmacokinetic parameters and that maximal killing is at 4 x MIC we recommend at least ceftazidime 2 g i.v . q8h.

J Antimicrob Chemother, 2002 Jan, 49(1), 121 - 8
Pharmacokinetics of ceftazidime in serum and peritoneal exudate during continuous versus intermittent administration to patients with severe intra-abdominal infections; Buijk SL et al.; Ceftazidime demonstrates time-dependent killing, which is maximal at 4 x or 5 x MIC for the organism, consequently continuous infusion (CI) has been proposed to ensure adequate ceftazidime concentrations for the entire course of therapy . Severe intra-abdominal infections (IAIs) require surgical or percutaneous drainage for management, and ceftazidime is frequently prescribed . Cardiovascular or metabolic changes and renal or liver dysfunction may alter drug pharmacokinetics during severe IAIs, and no data exist on concentrations of ceftazidime reached in the peritoneal fluid . The objectives here were to determine the pharmacokinetics of ceftazidime during continuous and intermittent administration in patients with severe IAIs, and to measure the concentrations of ceftazidime in the peritoneal exudate . Eighteen surgical patients with severe IAI and a creatinine clearance of >30 mL/min were studied . A non-randomized pilot study of six patients treated with CI alone was followed by a prospective, randomized comparative study of 12 patients . Pilot study patients received ceftazidime 1 g iv followed by a 4.5 g CI over 24 h . Randomized patients received either ceftazidime continuously as above or 1.5 g tds . Samples for pharmacokinetic analyses were collected on days 2 and 4 . Ceftazidime concentrations were determined by high-performance liquid chromatography . CI resulted in a mean serum concentration >40 mg/L and a T> 4 x MIC for most pathogens encountered in severe IAIs for >90% of the course of therapy in both serum and peritoneal exudate . Eight-hourly administration resulted in T> 4 x MIC for most pathogens encountered in severe IAIs for >90% of the dosing interval, but in peritoneal exudate for only 44% of the dosing interval . During CI, AUCs in the peritoneal exudate were c . 60% of the concomitant serum AUCs . In critically ill surgical patients with severe IAIs, CI of ceftazidime resulted in more favourable concentrations in serum and peritoneal exudate than 8-hourly bolus infusion.

J Antimicrob Chemother, 2002 Jan, 49(1), 49 - 54
Molecular analysis of chromosomally florfenicol-resistant Escherichia coli isolates from France and Germany; Doublet B et al.; The aim of this study was to analyse chromosomally florfenicol-resistant Escherichia coli isolates for their genetic relatedness, and also for the presence of the floR gene and its adjacent regions, in order to compare these regions with those associated with a floR gene located on a conjugative plasmid from E . coli . Twenty-two bovine E . coli from France and Germany were examined . Florfenicol resistance was determined by MIC determination . The presence of the floR gene was confirmed by hybridization and PCR analysis . The E . coli isolates were investigated by macrorestriction analysis . The 22 florfenicol-resistant E . coli (MICs 64->128 mg/L) differed in their BlnI macrorestriction patterns . Single or double copies of the floR gene were detected by hybridization on different-sized chromosomal EcoRI, BamHI and BglI fragments . The floR-flanking regions also proved to be variable as confirmed by hybridization experiments . The detection of chromosomal floR gene copies in unrelated E . coli isolates supplements the observations of floR genes on plasmids in E . coli and confirms their potential to integrate into the chromosome . The RFLPs of floR gene-carrying restriction fragments might suggest variable chromosomal integration sites.

J Antimicrob Chemother, 2002 Jan, 49(1), 41 - 7
A C-terminal 18 amino acid deletion in MarR in a clinical isolate of Escherichia coli reduces MarR binding properties and increases the MIC of ciprofloxacin; Notka F et al.; As described recently, the different degree of fluoroquinolone resistance in a pair of sequential clinical isolates of Escherichia coli was due to the increased expression of the regulatory gene marA as a consequence of an 18 amino acid C-terminal deletion in the repressor MarR (MarR Delta) . To further investigate the molecular mechanism of the loss of repressor function, we purified recombinant wild-type and mutated MarR, and tested their respective ability to form dimers and their specific DNA binding properties to the operator region marO . The dimerization capacity was analysed by non-reducing SDS-PAGE and by disuccinimidyl suberate-mediated cross-linking of the recombinant proteins . The binding of MarR was studied using the recombinant proteins and DNA probes containing the two identified binding sites in marO in the presence or absence of specific and non-specific DNA fragments . Dimerization of MarR Delta was reduced compared with MarR: the dimer portion was 33.8% (MarR) and 12.4% (MarR Delta) at a protein concentration of 10 IM . In mobility-shift assays MarR Delta showed a highly reduced complex formation . Footprinting analysis confirmed reduced binding of MarR Delta to its target sites, compared with MarR . The biochemical data are in full agreement with the crystal structure of MarR, which shows that the N- and C-terminal regions of MarR contribute to dimer formation . The data also indicate a major role of the MarR dimer as opposed to the monomer in DNA binding.

Antimicrob Agents Chemother, 2002 Jan, 46(1), 251 - 4
Antipneumococcal activity of BMS 284756 compared to those of six other agents; Pankuch GA et al.; Antipneumococcal activity of BMS 284756 was compared to those of six agents by MIC and time-kill methodologies . BMS 284756 had the lowest MICs compared to those of ciprofloxacin, levofloxacin, and moxifloxacin against quinolone-susceptible (< or =0.016 to 0.06 microg/ml) and quinolone-resistant (0.03 to 1 microg/ml) pneumococci . BMS 284756 was bactericidal against 11 of 12 strains at two times the MIC after 24 h.

Antimicrob Agents Chemother, 2002 Jan, 46(1), 62 - 8
In vitro activities of new and conventional antifungal agents against clinical Scedosporium isolates; Meletiadis J et al.; The susceptibilities of 13 clinical isolates of Scedosporium apiospermum and 55 clinical isolates of S . prolificans to new and conventional drugs belonging to three different classes of antifungal agents, the azoles (miconazole, itraconazole, voriconazole, UR-9825, posaconazole), the polyenes (amphotericin B, nystatin and liposomal nystatin), and allylamines (terbinafine), were studied by use of proposed standard M38-P of NCCLS . Low growth-inhibitory antifungal activities were found in vitro for most of the drugs tested against S . prolificans isolates, with the MICs at which 90% of isolates are inhibited (MIC(90)s) being >8 microg/ml; the MIC(90)s of voriconazole and UR-9825, however, were 4 microg/ml . S . apiospermum isolates were more susceptible in vitro, with the highest activity exhibited by voriconazole (MIC(90)s, 0.5 microg/ml), followed by miconazole (MIC(90)s, 1 microg/ml), UR-9825 and posaconazole (MIC(90)s, 2 microg/ml), and itraconazole (MIC(90)s, 4 microg/ml) . The MICs of terbinafine, amphotericin B, and the two formulations of nystatin (for which no statistically significant differences in antifungal activities were found for the two species) for S . apiospermum isolates were high . Cross-resistance was observed among all the azoles except posaconazole and among all the polyenes except the lipid formulation . A distribution analysis was performed with the MICs of each drug and for each species . Bimodal and skewed MIC distributions were obtained, and cutoffs indicating the borders of different MIC subpopulations of the distributions were determined on the basis of the normal plot technique . These cutoffs were in many cases reproducible between 48 and 72 h.

Antimicrob Agents Chemother, 2002 Jan, 46(1), 47 - 54
Mycobacterium smegmatis D-Alanine Racemase Mutants Are Not Dependent on D-Alanine for Growth; Chacon O et al.; Mycobacterium smegmatis is a fast-growing nonpathogenic species particularly useful in studying basic cellular processes of relevance to pathogenic mycobacteria . This study focused on the D-alanine racemase gene (alrA), which is involved in the synthesis of D-alanine, a basic component of peptidoglycan that forms the backbone of the cell wall . M . smegmatis alrA null mutants were generated by homologous recombination using a kanamycin resistance marker for insertional inactivation . Mutants were selected on Middlebrook medium supplemented with 50 mM D-alanine and 20 microg of kanamycin per ml . These mutants were also able to grow in standard and minimal media without D-alanine, giving rise to colonies with a drier appearance and more-raised borders than the wild-type strain . The viability of the mutants and independence of D-alanine for growth indicate that inactivation of alrA does not impose an auxotrophic requirement for D-alanine, suggesting the existence of a new pathway of D-alanine biosynthesis in M . smegmatis . Biochemical analysis demonstrated the absence of any detectable D-alanine racemase activity in the mutant strains . In addition, the alrA mutants displayed hypersusceptibility to the antimycobacterial agent D-cycloserine . The MIC of D-cycloserine for the mutant strain was 2.56 microg/ml, 30-fold less than that for the wild-type strain . Furthermore, this hypersusceptibility was confirmed by the bactericidal action of D-cycloserine on broth cultures . The kinetic of killing for the mutant strain followed the same pattern as that for the wild-type strain, but at a 30-fold-lower drug concentration . This effect does not involve a change in the permeability of the cell wall by this drug and is consistent with the identification of D-alanine racemase as a target of D-cycloserine . This outcome is of importance for the design of novel antituberculosis drugs targeting peptidoglycan biosynthesis in mycobacteria.

Antimicrob Agents Chemother, 2002 Jan, 46(1), 42 - 6
Antipneumococcal activity of ertapenem (MK-0826) compared to those of other agents; Pankuch GA et al.; The activities of ertapenem (MK-0826) and eight other agents against a range of penicillin-susceptible and -resistant pneumococci were tested by determination of MICs by the microdilution method and by the time-kill methodology . For 125 penicillin-susceptible, 74 penicillin-intermediate, and 86 penicillin-resistant pneumococci, the MICs at which 50% of isolates are inhibited (MIC(50)s) and MIC(90)s, as determined by the microdilution method, were as follows: for ertapenem, 0.016 and 0.03, 0.125 and 0.5, and 0.5 and 1.0 microg/ml for penicillin-susceptible, penicillin-intermediate, and penicillin-resistant pneumococci, respectively; for amoxicillin, < or =0.016 and 0.03, 0.25 and 1.0, and 2.0 and 2.0 microg/ml for penicillin-susceptible, penicillin-intermediate, and penicillin-resistant pneumococci, respectively; for cefprozil, 0.125 and 0.25, 1.0 and 8.0, and 16.0 and 16.0 microg/ml for penicillin-susceptible, penicillin-intermediate, and penicillin-resistant pneumococci, respectively; for cefepime, < or =0.016 and 0.06, 0.5 and 1.0, and 1.0 and 2.0 microg/ml for penicillin-susceptible, penicillin-intermediate, and penicillin-resistant pneumococci, respectively; for ceftriaxone, < or =0.016 and 0.06, 0.25 and 1.0, and 1.0 and 2.0 microg/ml for penicillin-susceptible, penicillin-intermediate, and penicillin-resistant pneumococci, respectively; for imipenem, < or =0.008 and < or =0.008, 0.03 and 0.25, and 0.25 and 0.25 microg/ml for penicillin-susceptible, penicillin-intermediate, and penicillin-resistant pneumococci, respectively; for meropenem, < or =0.008 and 0.016, 0.125 and 0.5, and 0.5 and 1.0 microg/ml for penicillin-susceptible, penicillin-intermediate, and penicillin-resistant pneumococci, respectively; and for clarithromycin, 1.0 and >32.0, 1.0 and >32.0, and >32.0 and >32.0 microg/ml for penicillin-susceptible, penicillin-intermediate, and penicillin-resistant pneumococci, respectively . For 64 strains for which quinolone MICs were increased (ciprofloxacin MICs, > or =4.0 microg/ml), the MIC(90) of ertapenem was 1.0 microg/ml and the MIC(90)s of the other beta-lactams tested and clarithromycin were >32.0 microg/ml . Against four penicillin-susceptible, four penicillin-intermediate, and four penicillin-resistant strains, testing by the time-kill methodology showed that ertapenem at two times the MIC was bacteriostatic (99% killing) after 12 h and bactericidal (99.9% killing) against all strains by 24 h, with 90% killing of all strains at two times the MIC after 6 h . At the MIC, ertapenem was bacteriostatic against all strains tested after 24 h . Although the bactericidal activity of imipenem at the MIC after 24 h was significantly greater than that of ertapenem, the kinetics of the two drugs at two times the MIC were similar after 24 h . The killing kinetics of clarithromycin were slower than those of ertapenem and other agents, with clarithromycin at two times the MIC having bactericidal activity against seven of eight macrolide-susceptible strains after 24 h.

Antimicrob Agents Chemother, 2002 Jan, 46(1), 12 - 23
Antifungal efficacy of caspofungin (MK-0991) in experimental pulmonary aspergillosis in persistently neutropenic rabbits: pharmacokinetics, drug disposition, and relationship to galactomannan antigenemia; Petraitiene R et al.; The antifungal efficacy, pharmacokinetics, and safety of caspofungin (CAS) were investigated in the treatment and prophylaxis of invasive pulmonary aspergillosis due to Aspergillus fumigatus in persistently neutropenic rabbits . Antifungal therapy consisted of 1, 3, or 6 mg of CAS/kg of body weight/day (CAS1, CAS3, and CAS6, respectively) or 1 mg of deoxycholate amphotericin B (AMB)/kg/day intravenously for 12 days starting 24 h after endotracheal inoculation . Prophylaxis (CAS1) was initiated 4 days before endotracheal inoculation . Rabbits treated with CAS had significant improvement in survival and reduction in organism-mediated pulmonary injury (OMPI) measured by pulmonary infarct score and total lung weight (P < 0.01) . However, animals treated with CAS demonstrated a paradoxical trend toward increased residual fungal burden (log CFU per gram) and increased serum galactomannan antigen index (GMI) despite improved survival . Rabbits receiving prophylactic CAS1 also showed significant improvement in survival and reduction in OMPI (P < 0.01), but there was no effect on residual fungal burden . In vitro tetrazolium salt hyphal damage assays and histologic studies demonstrated that CAS had concentration- and dose-dependent effects on hyphal structural integrity . In parallel with a decline in GMI, AMB significantly reduced the pulmonary tissue burden of A . fumigatus (P < or = 0.01) . The CAS1, CAS3, and CAS6 dose regimens demonstrated dose-proportional exposure and maintained drug levels in plasma above the MIC for the entire 24-h dosing interval at doses that were > or =3 mg/kg/day . As serial galactomannan antigen levels may be used for therapeutic monitoring, one should be aware that profoundly neutropenic patients receiving echinocandins for aspergillosis might have persistent galactomannan antigenemia despite clinical improvement . CAS improved survival, reduced pulmonary injury, and caused dose-dependent hyphal damage but with no reduction in residual fungal burden or galactomannan antigenemia in persistently neutropenic rabbits with invasive pulmonary aspergillosis.

Biotechnol Bioeng, 2001 Dec, 76(4), 391 - 4
A microbial chip combined with scanning electrochemical microscopy; Kaya T et al.; A microbial chip for bioassay was fabricated and its performance was characterized by scanning electrochemical microscopy (SECM) . The microbial chip was prepared by spotting a suspension of Escherichia coli on a polystyrene substrate by using a glass capillary pen . The respiration activity of the E . coli spot was imaged with SECM by mapping the oxygen concentration around the spot . The SECM images of the microbial chips clearly showed spots with lower reduction currents, indicating that E . coli in the spots uptake oxygen by respiration . The bactericidal effects of antibiotics (streptomycin and ampicillin) were measured using the E . coli-based microbial chip, and discussed in comparison with the minimum inhibitory concentration (MIC) determined by an agar plate dilution method .

Planta Med, 2001 Dec, 67(9), 873 - 5
Antifungal principles from Piper fulvescens; Freixa B et al.; Activity-guided fractionation of the dichloromethane extract from leaves of Piper fulvescens, using an agar overlay bioautographic method, led to the isolation of three antifungal neolignans identified as conocarpan, eupomatenoid 5 and eupomatenoid 6 . The minimal inhibitory concentration of these three neolignans against five fungi strains were determined . Conocarpan showed the widest activity, whereas eupomatenoid 6 was the most active against dermatophytes.

J Ethnopharmacol, 2002 Jan, 79(1), 57 - 67
The evaluation of forty-three plant species for in vitro antimycobacterial activities; isolation of active constituents from Psoralea corylifolia and Sanguinaria canadensis; Newton SM et al.; Extracts from forty-three plant species were selected on account of reported traditional uses for the treatment of TB and/or leprosy . These were assayed for antimycobacterial activities . A simple in vitro screening assay was employed using two model species of mycobacteria, M . aurum and M . smegmatis . Crude methanolic extracts from three of the plants, C . mukul, P . corylifolia and S . canadensis, were found to have significant antimycobacterial activity against M . aurum only (MIC=62.5 microg/ml) . Bioassay guided fractionation led to the isolation of two known benzophenanthridine alkaloids, sanguinarine (1) and chelerythrine (2), from the roots S . canadensis and the known phenolic meroterpene, bakuchiol (3) from the seeds of P . corylifolia . The fractionation of the resin of C . mukul lead to a decrease in antimycobacterial activity and hence further work was not pursued . Compound (2) was the most active against M . aurum and M . smegmatis (IC(50)=7.30 microg/ml {19.02 microM} and 29.0 microg/ml {75.56 microM}, respectively) . M . aurum was the most susceptible organism to all three compounds . No significant difference in antimycobacterial activity was observed when the two alkaloids were tested for activity in media of differing pH values . The activities of the pure compounds against M . aurum were comparable with those against M . bovis BCG with compound (2) being the most active (M . bovis BCG, IC(50)=14.3 microg/ml {37.3 microM}) . These results support the use of these plants in traditional medicine.

Acta Pharmacol Sin, 2001 Mar, 22(3), 210 - 4
Accumulation of ofloxacin and tosufloxacin in fluoroquinolone-resistant E coli; Xia PY et al.; AIM: To make sure whether there is a difference in mechanism existed in the resistant E coli strains accumulated hydrophilic fluoroquinolone ofloxacin and hydrophobic fluoroquinoline tosufloxacin . METHODS: Fluoroquinolone accumulation in bacteria and effect of active efflux were measured by fluorescence method . Analysis of outer membrane proteins was made by SDS-PAGE . E coli strains included JF701 and JF703 that are OmpC- or OmpF-deficient mutants of E coli K-12 respectively, and the susceptible strain Ecs and its in-vitro selected resistant strains R2, R256, and clinical resistant isolates R5, R6 . RESULTS: Ecs accumulated ofloxacin almost at the same concentration as JF701, but JF703 did about 1/2 of that lower than JF701 . However, four resistant strains accumulated ofloxacin about 5 to 7-fold lower than those susceptible strains . On the other hand, there was no significant difference for the accumulation of tosufloxacin between fluoroquinolone-resistant and -susceptible strains . After addition of proton ionophore DNP for 5 min and 10 min, the accumulation of tosufloxacin slowly decreased in E coli strains, whereas the accumulation of ofloxacin was increased, especially in the resistant strains . A good relevance exists between the accumulation increment of ofloxacin and its MIC for each E coli strain after addition of DNP for 5 min and 10 min (r=0.9623 and 0.8006 respectively) . Furthermore, both OmpF and OmpC in Ecs, OmpF-deficiency in R2, R256 and OmpC-deficiency in R5, R6 were observed . CONCLUSION: The accumulation of ofloxacin other than tosufloxacin could be reduced by OmpF-deficiency and active efflux, and the latter may be an important factor in the development of resistance to hydrophilic fluoroquinolone in E coli.

J Cataract Refract Surg, 2001 Dec, 27(12), 1969 - 74
Human aqueous humor levels of oral ciprofloxacin, levofloxacin, and moxifloxacin; Garcia-Saenz MC et al.; PURPOSE: To evaluate the penetration of ciprofloxacin, levofloxacin, and moxifloxacin into the aqueous humor after oral administration . SETTING: Alcorcon Hospital, Madrid, Spain . METHODS: Forty-two patients having cataract surgery were randomly divided into 3 groups the day before surgery . The first group received 2 oral 500 mg doses of ciprofloxacin at 12-hour intervals . The second group received a single oral 500 mg dose of levofloxacin . The third group received a single oral 400 mg dose of moxifloxacin . At the time of surgery, 0.1 mL aqueous fluid was aspirated from the anterior chamber just before the operation and immediately stored at -80 degrees C . Drug concentrations were measured using a biological assay . RESULTS: The mean aqueous level of ciprofloxacin was 0.50 microg/mL +/- 0.25 (SD); of levofloxacin, 1.50 +/- 0.50 microg/mL; and of moxifloxacin, 2.33 +/- 0.85 microg/mL . The mean aqueous levels of levofloxacin and moxifloxacin were above the 90% minimum inhibitory concentration for most of the common microorganisms that cause endophthalmitis . CONCLUSIONS: Therapeutic concentrations of fluoroquinolones, mainly levofloxacin and moxifloxacin, were reached with oral administration . These antibiotics may be effective for prophylaxis and adjuvant therapy of bacterial endophthalmitis.

Int J Radiat Oncol Biol Phys, 2001 Dec 1, 51(5), 1264 - 70
Pathologic downstaging of T3-4Nx rectal cancer after chemoradiation: 5-fluorouracil vs . Tegafur; Calvo FA et al.; PURPOSE: To describe downstaging effects in locally advanced rectal cancer induced by 2 fluopirimidine radiosensitizing agents given through different routes in conjunction with preoperative radiotherapy . METHODS AND MATERIALS: From March 1995 to December 1999, two consecutive groups of patients with cT3-4Nx rectal cancer (94% CT scan, 71% endorectal ultrasound) were treated with either (1) 45-50 Gy (1.8 Gy/day, 25 fractions) and 5-fluorouracil (5-FU) (500-1,000 mg/m2 by 24-h continuous i.v . infusion on Days 1-4 and 21-25) or (2) oral Tegafur (1,200 mg/day on Days 1-35, including weekends) . Surgery was performed 4 to 6 weeks after the completion of chemoradiation . RESULTS: The total T downstaging rate was 46% in the 5-FU group and 53% in the Tegafur group . Subcategories were downstaged by the sensitizing agents (5-FU vs . Tegafur) as follows: pT0-1, 14% vs . 23%; pT2, 32% vs . 32%; pT3, 49% vs . 37%; pT4, 5% vs . 7%; and N(0), 74% vs . 86% . Analysis of residual malignant disease in the specimen discriminated mic/mac subgroups (mic: <20% of microscopic cancer residue), with evident superior downstaging effects in the Tegafur-treated group: pTmic 23% vs . 58% (p = 0.002) . CONCLUSIONS: When administered concurrent with pelvic irradiation, oral Tegafur induced downstaging rates in both T and N categories superior to those induced by intermediate doses of 5-FU by continuous i.v . infusion . In this pilot experience, oral Tegafur reproduced the characteristics of downstaging described previously when full doses of 5-FU have been combined with radiotherapy.

CLAO J, 2001 Oct, 27(4), 212 - 5
Disposable 1-day Acuvue contact lenses for the delivery of lomefloxacin to rabbits' eyes; Tian X et al.; PURPOSE: To investigate the ability of a disposable soft contact lens (1-Day Acuvue) to deliver lomefloxacin, a fluoroquinolones antibiotic, into the cornea and aqueous humor of rabbits . METHODS: Experiments were conducted on adult albino rabbits . 1-Day Acuvue lenses were immersed for 1 hour in commercially-available lomefloxacin eye solution and then placed on the cornea . After 0.5, 2, 4,6, and 8 hours, the animals were killed and the lenses were removed and placed into a 20 mL saline bath . Corneal tissue and aqueous humor samples were also obtained at these times . The release of lomefloxacin from the lenses was calculated by measuring the amount of drug remaining in the lenses after removal from the rabbit eyes . The concentration of lomefloxacin in the cornea and anterior chamber following the wearing of lomefloxacin-loaded lenses was compared with the concentrations following frequent-drop therapy (one drop of lomefloxacin hourly for 8 hours) . RESULTS: In vivo, lomefloxacin was released from 1-Day Acuvue lenses gradually over 8 hours . The cornea and aqueous humor levels in the eyes following the wear of lomefloxacin-loaded lenses were significantly higher than those achieved by frequent-drop therapy . The drug levels in the cornea and aqueous humor generally remained above the 90% minimal inhibitory concentration for 8 hours in the 1-Day Acuvue lens group . CONCLUSIONS: 1-Day Acuvue contact lenses soaked in 0.3% lomefloxacin can release sufficient amounts of lomefloxacin and in this study produced higher levels in both the cornea and aqueous humor than that achieved by frequent-drop therapy for up to 8 hours . We conclude that 1-Day Acuvue contact lens can be used as a drug delivery system for lomefloxacin.

J Clin Microbiol, 2001 Dec, 39(12), 4256 - 63
Comparison of spectrophotometric and visual readings of NCCLS method and evaluation of a colorimetric method based on reduction of a soluble tetrazolium salt, 2,3-bis {2-methoxy-4-nitro-5-{(sulfenylamino) carbonyl}-2H-tetrazolium-hydroxide}, for antifungal susceptibility testing of Aspergillus species; Meletiadis J et al.; The susceptibilities of 25 clinical isolates of various Aspergillus species (Aspergillus fumigatus, A . flavus, A . terreus, A . ustus, and A . nidulans) to itraconazole (ITC) and amphotericin B (AMB) were determined using the standard proposed by NCCLS for antifungal susceptibility testing of{filamentous fungi, a modification of this method using spectrophotometric readings, and a colorimetric method using the tetrazolium salt 2,3-bis {2-methoxy-4-nitro-5-{(sulfenylamino) carbonyl}-2H-tetrazolium-hydroxide} (XTT) . Five MIC end points for ITC (MIC-0, no visible growth or <or=5% the growth control value {GC}; MIC-1, slight growth or 6 to 25% the GC; MIC-2, prominent reduction in growth or 26 to 50% the GC; MIC-3, slight reduction in growth or 51 to 75% the GC; and MIC-4, no reduction in growth or 76 to 100% the GC) and one for AMB (MIC-0) were determined visually by four observers and spectrophotometrically . The intraexperimental (between the observers) and interexperimental (between the experiments) levels of agreement of the NCCLS and XTT methods exceeded 95% for MIC-0 of AMB and MIC-0 and MIC-1 of ITC . The MIC-2 of ITC showed lower reproducibility, although spectrophotometric reading and/or incubation for 48 h increased the interexperimental reproducibility from 85 to >93% . Between visual and spectrophotometric readings, high levels of agreement were found for AMB (approximately 97%) and MIC-1 (approximately 92%) and MIC-2 (approximately 88%) of ITC . Poor agreement was found for MIC-0 of ITC (51% after 24 h), since the spectrophotometric readings resulted in higher MIC-0 values than the visual readings . The agreement was increased to 98% by shifting the threshold level of MIC-0 from 5 to 10% relative optical density and by establishing an optical density of greater than 0.1 for the GC as the validation criterion . No statistically significant differences were found between the NCCLS method and the XTT method, with the levels of agreement exceeding 97% for MIC-0 of AMB and 83% for MIC-0, MIC-1, and MIC-2 of ITC . The XTT method and spectrophotometric readings can increase the sensitivity and the precision, respectively, of in vitro susceptibility testing of Aspergillus species.

Acta Psychiatr Scand, 2001 Oct, 104(4), 264 - 72
Internal consistency, intercriterion overlap and diagnostic efficiency of criteria sets for DSM-IV schizotypal, borderline, avoidant and obsessive-compulsive personality disorders; Grilo CM et al.; OBJECTIVE: To evaluate performance characteristics of DSM-IV Personality Disorders (PDs) criteria . METHOD: Six hundred and sixty-eight adults recruited for the Collaborative Longitudinal Personality Disorders Study (CLPS) were assessed with diagnostic interviews . RESULTS: Within-category inter-relatedness was evaluated by Cronbach's alpha and median intercriterion correlations (MIC) . Cronbach's alpha ranged from 0.47 to 0.87 (median=0.71); seven of the 10 PDs had alphas greater than 0.70 . Between-category criterion overlap was evaluated by "inter-category" intercriterion correlations between all PD pairs (ICMIC) . ICMIC values (median=0.08) were lower than MIC values (median=0.23) . Diagnostic efficiency statistics (sensitivity, specificity, positive predictive power and negative predictive power were calculated for schizotypal, borderline, avoidant and obsessive-compulsive PDs . CONCLUSION: DSM-IV PD criteria sets have some convergent validity and discriminant validity: criteria for individual PDs correlate better with each other than with criteria for other PDs . Diagnostic efficiency statistics provide guidance regarding usefulness of criteria for inclusion or exclusion.

J Nat Prod, 2001 Nov, 64(11), 1463 - 4
Inhibition of Mycobacterium tuberculosis growth by saringosterol from Lessonia nigrescens; Wachter GA et al.; Assay-guided fractionation of an antitubercular extract obtained from Lessonia nigrescens yielded the phytosterol saringosterol as its active component . No appreciable toxicity against Vero cells was observed for this compound . Saringosterol was also synthesized by oxidation of fucosterol . The MIC values for antitubercular activity of saringosterol and its 24S and 24R epimers were determined as 0.25, 1, and 0.125 microg/mL.

Lung Cancer, 2001 Dec, 34(3), 351 - 61
Topographical analysis of p53 expression and DNA ploidy in early bronchial squamous cell carcinoma and preneoplastic lesions; Inage Y et al.; The significance of p53 mutations and DNA aneuploidy in carcinoma cells has been investigated on the basis of a multi-step development theory of carcinogenesis . It has, however, not been determined whether these alterations can be used as diagnostic markers for the early detection of bronchial squamous cell carcinoma (BSqCC) . To address this problem, we topographically investigated p53 alterations and DNA aneuploidy in 24 X-ray-negative, early BSqCC patients with various preneoplastic lesions and in 25 non-carcinoma patients with preneoplastic lesions . Bronchial lesions (n=88) were morphologically classified as hyperplasia (HP, n=5), squamous metaplasia (SM, n=23), low-grade dysplasia (LGD, n=14), high-grade dysplasia (HGD, n=11), intraepithelial carcinoma including 'carcinoma in situ' (CIS) (IEC, n=15), and microinvasive carcinoma (MIC, n=20) . Immunohistochemistry for the p53 protein and image cytometry for DNA ploidy detection were performed in serial sections of each lesion . Overexpression of p53 protein was detected in 36, 73, and 65% of the HGD, IEC, and MIC lesions, respectively . Aneuploid DNA profiles were found only in carcinoma lesions, 33% in IEC and 85% in MIC . The topographical analysis revealed two types of early BSqCCs, one with adjacent preneoplastic lesions (sequential type, n=8) and another without such lesions (de novo type, n=16) . The p53 protein was frequently overexpressed in both types (sequential type, 79%; de novo type, 62%) . In the sequential type, however, the p53 protein was overexpressed in HGD lesions that were directly adjacent to p53-overexpressing carcinoma lesions without exception . The present topographical study suggests that p53 mutations play an important role in the carcinogenesis of BSqCC and that p53-overexpressing HGD lesions in sequential types should be regarded as 'truly' preneoplastic lesions that actually develop into carcinomas . In addition, our study demonstrated that DNA aneuploidy might occur at times after p53 alteration with increasing frequency, as invasive growth begins . Such combination analysis of p53 immunohistochemistry and nuclear DNA ploidy in routine histology may contribute to estimates of malignant potential in preneoplastic and intraepithelial squamous lesions and provide additional information for early detection of BSqCC.

J Agric Food Chem, 2001 Nov, 49(11), 5251 - 7
N-heterocyclic derivatives of 2,4-dihydroxybenzcarbothioamide as antimycotic agents; Matysiak J et al.; N-heterocyclic derivatives of 2,4-dihydroxybenzcarbothioamide were synthesized from sulfinylbis(2,4-dihydroxybenzenethioyl) and commercially available heterocyclic amines . The composition and chemical structures were confirmed by IR, (1)H NMR, EI-MS, and elemental analysis . For the estimation of potential activity in vitro the MIC values against 15 strains of dermatophytes, yeasts, and molds were determined . The strongest fungistatic potency was found for N-5'-(3'-oxobenzfurylidyne)-2,4-dihydroxybenzcarbothioamide in relation to all tested dermatophyte strains with MIC = 0.48-0.98 microg/mL . On the basis of the spectroscopic data the influence of N-heterocyclic substitution on antimycotic activity is discussed.

Pharmacotherapy, 2001 Nov, 21(11 Pt 2), 302S - 318S
What do we really know about antibiotic pharmacodynamics?
Gunderson BW, Ross GH, Ibrahim KH, Rotschafer JC.
Antibiotic pharmacodynamics is an evolving science that focuses on the relationship between drug concentration and pharmacologic effect, which is an antibiotic-induced bacterial death that also can manifest as an adverse drug reaction . The pharmacologic action of antibiotics usually can be described as concentration dependent or independent, although such classifications are highly reliant on the specific antibiotic and bacterial pathogen being studied . Quantitative pharmacodynamic parameters, such as ratio of the area under the concentration-time curve during a 24-hour dosing period to minimum inhibitory concentration (AUC0-24:MIC), ratio of maximum serum antibiotic concentration to MIC (Cmax:MIC), and duration of time that antibiotic concentrations exceed MIC (T>MIC), have been proposed as likely predictors of clinical and microbiologic success or failure for different pairings of antibiotic and bacteria . Thus far, most pharmacodynamic data reported have focused on fluoroquinolones, but work has been conducted on vancomycin, beta-lactams, macrolides, aminoglycosides, and other antibiotics . Despite the development of a number of different pharmacodynamic modeling systems, remarkable agreement exists between in vitro, animal, and limited human data . Although still somewhat premature and requiring additional clinical validation, antibiotic pharmacodynamics will likely advance on four fronts: the science should prove to be extremely useful and represent a cost-effective and efficient method to help develop new antibiotics; formulary committees will likely use pharmacodynamic parameters to assist in differentiating antibiotics of the same chemical class in making antibiotic formulary selections; pharmacodynamic principles will likely be used to design optimal antibiotic strategies for patients with severe infections; and limited data to date suggest that the application of pharmacodynamic concepts may limit or prevent the development of antibiotic resistance . The study of antibiotic pharmacodynamics appears to hold great promise and will likely become a routine part of our daily clinical practices.

Mod Pathol, 2001 Nov, 14(11), 1175 - 82
Differentiating lymphoblastic lymphoma and Ewing's sarcoma: lymphocyte markers and gene rearrangement; Ozdemirli M et al.; We encountered a child with an intraosseous small round cell tumor that was negative for LCA, CD20 (L26), and CD3 and positive for vimentin, CD99 (MIC-2), and periodic acid-Schiff . The tumor exhibited rosette-like formations . This case was initially interpreted as Ewing's sarcoma (ES); however, additional studies revealed positivity for CD79a, CD43, and TdT expression, and an immunoglobulin heavy chain gene rearrangement (IgH-R) by polymerase chain reaction (PCR) established this to be a precursor B-lymphoblastic lymphoma . Because the differential diagnosis of ES and lymphoblastic lymphoma can be difficult and the differential diagnostic value of leukocyte antigens and immunoglobulin heavy chain gene rearrangement studies have not been fully evaluated, we conducted a more extensive investigation on 33 (21 soft tissue and 12 intraosseous) ES cases . Cases were retrieved from the files of the Department of Pathology at Georgetown University and from the Soft Tissue Registry of the Armed Forces Institute of Pathology . The cases were studied by light microscopy, immunohistochemistry, and PCR for IgH-R and T cell receptor gamma chain gene rearrangement (Tgamma-R) . There were 17 females and 16 males; the mean age was 29.3 years . Locations included the extremities (n = 17) and trunk (n = 16) . All cases fit the ES spectrum by light microscopy and immunohistochemistry, as previously determined, and were negative for lymphoid markers (LCA, CD3, CD20, CD43, CD79a, and TdT), CD10 and CD34 . CD99 was positive in 31/33 and bcl-2 was weakly positive in 13/33 cases . All 21 cases studied for gene rearrangements by PCR were negative for IgH-R and Tgamma-R . Distinction of intraosseous lymphoblastic lymphoma from ES may be difficult because lymphomas may occasionally exhibit unexpected morphologic and immunophenotypic properties including LCA, CD3 and CD20 negativity and cytokeratin positivity . Additional analysis using CD79a, CD43, TdT, and PCR should be performed to avoid misdiagnosis . True ES is negative for lymphoid markers including CD79a, CD43, and TdT, as well as for IgH-R and Tgamma-R.

Rev Esp Quimioter, 2001 Jun, 14(2), 191 - 7
{In vitro antifungal susceptibility of dematiaceous filamentous fungi using the E-test}; Vivas JR et al.; The usefulness of the E-test((R) )has been studied to determine the in vitro susceptibility of 52 isolates of 17 species to five antifungal drugs: amphotericin B, 5-flucytosine, ketoconazole, itraconazole and fluconazole . Minimal inhibitory concentrations were determined following the manufacturers' instructions, except in the preparation of the inoculum . In this case a spectrophotometric method was used to obtain 1-5 ' 10(6) CFU/ml . Two different culture media were included: casitone-agar and RPMI 1640 agar . Most isolates showed clear growth in both media after 96 h of incubation at 37 degrees C . The species showed low MIC concentrations to ketoconazole and itraconazole . Only 55.8% of isolates showed MICs </= 2 mg/l to amphotericin B and 98% of strains presented MICs of 32 mg/l for 5-flucytosine . With the exception of one isolate, the remaining were resistant to fluconazole with MIC values 256 mg/l . No differences were found between the MICs for the two media, but values were easier to read in casitone-agar . These results suggest that itraconazole and ketoconazole should be the antifungals of first choice for treating infections due to the evaluated species . This study showed that the E-test((R) )appears to be a suitable method for testing the susceptibility of dematiaceous filamentous fungi to antifungals.

Tissue Antigens, 2001 Sep, 58(3), 159 - 65
Genomic analysis of MIC genes in rhesus macaques; Seo JW et al.; MIC genes map to the major histocompatibility complex (MHC) and are distantly related to MHC class I genes . Recently, MICA/MICB-like genes have been described in nonhuman primates . In Macaca mulatta, three MICA/B-like genes could be identified: Mamu-MIC1, Mamu-MIC2, and Mamu-MIC3 . We show here the isolation and characterization of rhesus macaque cosmid clones which carry the Mamu-MIC2 and Mamu-MIC3 genes . Neither the MIC2- and MIC3-coding sequences nor respective flanking sequences can be aligned unambiguously to either the human HLA-MICA or -MICB subregions, although MIC2 was found at a similar distance to the BAT1 gene as known for MICB in human . Thus, the characteristics allowing for a classification of primate MIC genes as being of the MICA or MICB types appear to have evolved after the separation of humans and rhesus monkeys from a common ancestor . Furthermore, also Mamu-MICD-containing cosmids could be isolated . In contrast to Mamu-MIC2 and Mamu-MIC3, the Mamu-MICD gene and its flanking sequences are highly conserved and orthologous to the human MICD subregion.

Mol Microbiol, 2001 Oct, 42(2), 309 - 17
Arylamine N-acetyltransferase of Mycobacterium tuberculosis is a polymorphic enzyme and a site of isoniazid metabolism; Upton AM et al.; Arylamine N-acetyltransferases (NATs; E.C 2.3.1.5) N-acetylate arylhydralazine and arylamine substrates using acetyl coenzyme A . Human NAT2 acetylates and inactivates the antituberculosis drug, isoniazid (INH), and is polymorphic . We previously demonstrated that there is a homologue of human NAT2 in Mycobacterium tuberculosis, whose product N-acetylates INH in vitro . We now demonstrate that the nat gene is expressed in M . tuberculosis and M . bovis Bacille Calmette-Guerin (BCG), using reverse transcription-polymerase chain reaction and Western blotting . The NAT protein is active in M . bovis BCG in vivo, as detected by the presence of N-acetyl INH in M . bovis BCG lysates grown in INH . Sequence analysis of the M . tuberculosis nat coding region reveals a single nucleotide polymorphism in 18% of a random cohort of M . tuberculosis clinical isolates, conferring a G to R change . The recombinant mutant protein appears less stable than the wild type, and has an apparent affinity for INH of 10-fold less than the wild type . Modelling the change in M . tuberculosis NAT shows that the G to R change is close to the active site, and supports the experimental findings . Minimum inhibitory concentration data suggest that this polymorphism in nat is linked to low-level changes in the INH susceptibility of M . tuberculosis clinical isolates.

J Clin Endocrinol Metab, 2001 Nov, 86(11), 5152 - 8
Galectin-3 as a presurgical immunocytodiagnostic marker of minimally invasive follicular thyroid carcinoma; Saggiorato E et al.; Thyroid nodules are a common occurrence in the general population, but only a small number of them are eventually diagnosed as cancers . Fine-needle aspiration biopsy (FNAB) is the most accurate and cost-effective method for the presurgical management of thyroid nodules, but it misses the differential diagnosis between thyroid follicular adenomas and follicular carcinomas . Among them, minimally invasive follicular carcinoma (MIC), also defined as encapsulated tumor, only differs from follicular adenoma for the exhibition of minimal, but entire thickness, infiltration of the capsule and/or vascular invasion . This feature cannot be assessed in FNAB and can occasionally be hard to recognize in surgical specimens . As reported in several studies, galectin-3 is a reliable marker of thyroid malignancy, but no data are available on MICs . We analyzed the immunohistochemical expression of galectin-3 in 17 MICs and 52 follicular adenomas in both preoperative paraffin-embedded cytological human thyroid sediments (cell blocks) obtained by FNAB and in the corresponding surgical specimens . Among the MICs, all surgical samples showed galectin-3 immunoreactivity in the cytoplasm, whereas 16 of 17 corresponding FNAB cell blocks were positive . No evidence of cytoplasmic galectin-3 expression was observed in 48 of 52 adenomas in both cell blocks and histological tissues . These findings indicate that galectin-3 is a reliable presurgical molecular marker of MIC, improving the accuracy of conventional FNAB . It also proves to be useful in the histopathological assessment of resected tumors having suspected malignant features.

J Ethnopharmacol, 2001 Dec, 78(2-3), 213 - 6
Inhibition of drug-sensitive and drug-resistant strains of Mycobacterium tuberculosis by diospyrin, isolated from Euclea natalensis; Lall N et al.; The binaphthoquinoid, diospyrin, was isolated from Euclea natalensis A.DC., and evaluated for its activity against drug-sensitive and drug-resistant strains of Mycobacterium tuberculosis . The minimal inhibitory concentration (MIC) of diospyrin was found to be 100 microg/ml for all the M . tuberculosis strains.

Clin Infect Dis, 2001 Dec 1, 33(11), 1910 - 3 Epub 2001 Oct 23.
Emergence of resistance to fluconazole as a cause of failure during treatment of histoplasmosis in patients with acquired immunodeficiency disease syndrome; Wheat LJ et al.; In sequential clinical trials of treatment for histoplasmosis in patients with acquired immunodeficiency syndrome, therapy with fluconazole failed in a higher proportion of patients than did therapy with itraconazole . To determine the cause for failure with fluconazole, antifungal susceptibility testing that used modified National Committee on Clinical Laboratory Standards procedures was performed on all baseline and failure isolates . Failure occurred more frequently in patients with baseline isolates with fluconazole minimum inhibitory concentrations (MICs) > or =5 microg/mL versus lower MICs; 29% versus 3%, respectively . There was at least a 4-fold increase in fluconazole MIC in the isolates from 10 (59%) of 17 patients for whom paired pretreatment and failure or relapse isolates were available . Cross-resistance to itraconazole was not seen . In conclusion, fluconazole is less active than itraconazole for Histoplasma capsulatum and induces resistance during therapy, which accounted for treatment failure in some patients.

Int J Antimicrob Agents, 2001 Oct, 18(4), 387 - 90
Primary resistance to clarithromycin in clinical strains of Helicobacter pylori isolated from children in Poland; Dzierzanowska-Fangrat K et al.; Helicobacter pylori resistance to clarithromycin is an important factor in the failure of eradication therapy . The resistance results from point mutations in the 23S rRNA gene of H . pylori . The prevalence of primary resistance of H . pylori to clarithromycin in children and mutations associated with resistance were studied and it was found that 23.5% (23/98) of H . pylori strains isolated in our hospital during 1998-2000 were resistant to clarithromycin . The primary resistance was mainly caused by an A2143G mutation, but the isolates with an A2142G mutation had higher MICs for clarithromycin compared with those with an A2143G mutation: median MIC 256 versus 16 mg/l . Comparison of our data with previous results showed that the prevalence of H . pylori resistance to clarithromycin in children has increased in Poland over the last three years, however the difference was not significant (23.5 vs . 17%, P=0.22).

Int J Antimicrob Agents, 2001 Oct, 18(4), 335 - 40
Tissue and serum concentrations of levofloxacin in orthopaedic patients; von Baum H et al.; The level of levofloxacin was determined in serum, bone and several tissues after a single dose of 500 mg i.v . Twenty-one patients (mean age: 56.8 years) undergoing bone surgery (nine patients) or surgical debridement of a decubitus ulcer (12 patients) who received levofloxacin as perioperative prophylaxis were included in the study . During surgery, blood and tissue samples were obtained approximately 1.5 h (range 40-210 min) postdosing . Levofloxacin concentrations in 87 specimens including 21 serum samples were determined using high-performance liquid chromatography (HPLC) . The mean serum concentration at 1.5 h was 8.6+/-2.3 microg/ml . Concentrations above the MIC of common pathogens were reached in all tissues during the collection period with a maximum in skin samples (19.9+/-9.9 microg/g) followed by wound tissue and granulation tissue with 17.3+/-6.5 and 13.7+/-6.4 microg/g respectively . In muscle and fatty tissue mean levofloxacin concentrations of 8.0+/-0.9 and 4.0+/-2.2 microg/g were attained . Mean levels in cancellous bone were 6.6+/-3.6 microg/g, lowest levels were measured in cortical bone (2.8+/-1.1 microg/g) . Twenty-two different pathogens were cultivated from the lesions of 11 of 12 patients with pressure ulcers . MIC values for levofloxacin were determined and compared with the corresponding tissue concentrations . Levofloxacin may be useful for perioperative prophylaxis and treatment in orthopaedic patients due to its good tissue penetration.

Diagn Microbiol Infect Dis, 2001 Sep-Oct, 41(1-2), 79 - 82
In vitro susceptibility study of BMS-284756 against Legionella species; Dubois J et al.; Legionella organisms are often associated with respiratory infections, and Legionella pneumonia results in significant mortality unless it is promptly and effectively treated . The present study was undertaken to compare the in vitro activity of BMS-284756 (T-3811ME), a novel des-F(6)-quinolone, against Legionella species versus the activity of other fluoroquinolones (levofloxacin, moxifloxacin, and ciprofloxacin) and of the macrolides erythromycin, clarithromycin, and azithromycin . The most potent agents tested against Legionella pneumophila serogroup 1, the largest group tested, were BMS-284756, moxifloxacin, and levofloxacin (MIC(90) = 0.016 mg/L) . The MIC(90) range for BMS-284756 was 0.008-0.03 mg/L against the total panel of L pneumophila serogroups 1-9 and 12, with the lowest MIC(90) observed for serogroup 7 and the highest for serogroup 2 . BMS-284756 was one of the most potent agents tested against isolates of L micdadei, L longbeachae, and other Legionella species (MIC(90) range: 0.008-0.06 mg/L) . These results and the high intrinsic activity of BMS-284756 against other respiratory pathogens support its use as empiric monotherapy for a wide range of respiratory infections.

Acta Otorhinolaryngol Belg, 2001, 55(3), 259 - 64
Amoxicillin/clavulanate in chronic rhinosinusitis: tissue and serum distribution; Passali D et al.; Amoxicillin/clavulanate in chronic rhinosinusitis: tissue and serum distribution . OBJECTIVE: The aim of the present study is to determine the concentrations of coamoxiclav in the sinusal mucosa of patients undergoing surgery for chronic sinusitis in comparison to serum levels after single oral administration . METHODOLOGY: 24 patients affected by chronic sinusitis, undergoing sinus surgery, were divided into three groups receiving an oral dose of 1 g of coamoxiclav (875 mg amoxicillin, 125 g of clavulanic acid, 7:1 ratio) at 2 h (first group), 4 h (second group) and 6 h (third group) before surgery . The mean concentration of amoxicillin and clavulanic acid were determined biologically in serum and in tissues . RESULTS: The highest concentrations of coamoxiclav both in serum and tissues were observed in the group which received the antibiotic 2 hours before surgery . However the tissue levels of both amoxicillin and clavulanic acid in the time period within 2-6 h after administration were higher than the Minimum Inhibitory Concentration (MIC) for the most frequent causative pathogens of sinus bacterial infections . CONCLUSIONS: Since penicillins need to maintain concentrations above the MIC for at least 40 to 60% of the interval time between administrations to be potentially effective, the Authors concluded that since both amoxicillin and clavulanic acid spread well in the ENT tissues, 1 g twice a day of the combination seems to be clinically effective even in patients suffering from acute episodes of chronic rhinosinusitis.

J Clin Microbiol, 2001 Nov, 39(11), 4208 - 9
In vitro activity of posaconazole against clinical isolates of dermatophytes; Barchiesi F et al.; A broth macrodilution method following the recommendations established by the National Committee for Clinical Laboratory Standards was used to compare the in vitro activity of posaconazole (PCZ) with that of itraconazole (ITC) against 30 clinical isolates of dermatophytes belonging to six different species . In terms of MICs, PCZ showed an activity equal to that of ITC . MICs of PCZ at which 50% (MIC(50)) and 90% (MIC(90)) of the isolates were inhibited were 0.5 and > 4.0 microg/ml, respectively . The MIC(50) and MIC(90) of ITC were 1.0 and > 4.0 microg/ml, respectively . However, PCZ showed a more potent fungicidal activity than that of ITC against isolates belonging to the genus Microsporum (P = 0.03) . PCZ merits further investigation as a potentially useful agent for treatment of dermatophytosis.

J Clin Microbiol, 2001 Nov, 39(11), 4103 - 10
Comparison of methods for Identification of Mycobacterium abscessus and M . chelonae isolates; Yakrus MA et al.; Mycobacterium abscessus and Mycobacterium chelonae are two closely related species that are often not distinguished by clinical laboratories despite the fact they cause diseases requiring different treatment regimens . Multilocus enzyme electrophoresis, PCR-restriction fragment length polymorphism analysis of the 65-kDa heat shock protein gene, biochemical tests, and high-performance liquid chromatography of mycolic acids were used to identify 75 isolates as either M . abscessus or M . chelonae that were originally submitted for drug susceptibility testing . Only 36 of these isolates were submitted with an identification at the species level . Using the above methods, 46 of the isolates were found to be M . abscessus and 29 were identified as M . chelonae . Eight isolates originally submitted as M . chelonae were identified as M . abscessus, and one isolate submitted as M . abscessus was found to be M . chelonae . The four identification methods were in agreement in identifying 74 of the 75 isolates . In drug susceptibility testing, all isolates of M . abscessus exhibited resistance to tobramycin (MIC of 8 to > or =16 microg/ml), while all isolates of M . chelonae were susceptible to this drug (MIC of < or = 4 microg/ml) . The results suggest that once an identification method is selected, clinical laboratories should be able to easily identify isolates of M . abscessus and M . chelonae.

J Antimicrob Chemother, 2001 Nov, 48(5), 705 - 7
Antimycobacterial activity of N1-{1-{3-aryl-1-(pyridin-2-, 3- or 4-yl)-3-oxo} propyl}-2-pyridinecarboxamidrazones; Banfi E et al.; Infections caused by non-tuberculous mycobacteria and multidrug-resistant Mycobacterium tuberculosis are difficult to treat, and so new compounds potentially active against these bacteria are being sought . A series of 2-pyridinecarboxamidrazone derivatives, recently synthesized, have been evaluated for their inhibitory activity against 17 Mycobacterium avium isolates; the agar dilution method showed different degrees of susceptibility to the new molecules . Four molecules, three of which are chlorine derivatives, inhibited 94% of the strains tested with an MIC of 32 mg/L . These data indicate that these new pyridine-2-carboxamidrazones merit further study as antimycobacterial agents.

Biosens Bioelectron, 2001 Dec, 16(9-12), 995 - 1000
Microbial corrosion monitoring by an amperometric microbial biosensor developed using whole cell of Pseudomonas sp; Dubey RS et al.; A microbial biosensor was developed for monitoring microbiologically influenced corrosion (MIC) of metallic materials in industrial systems . The Pseudomonas sp . isolated from corroded metal surface was immobilized on acetylcellulose membrane and its respiratory activity was estimated by measuring oxygen consumption . The microbial biosensor was used for the measurement of sulfuric acid in a batch culture medium contaminated by microorganisms . A linear relationship between the microbial sensor response and the concentration of sulfuric acid was observed . The response time of biosensor was 5 min and was dependent on the immobilized cell loading of Pseudomonas sp., pH, temperature and corrosive environments . The microbial biosensor response was stable, reproducible and specific for sensing of sulfur oxidizing bacterial activity.

J Mol Evol, 2001 Dec, 53(6), 642 - 50
Genomic and phylogenetic analysis of the human CD1 and HLA class I multicopy genes; Kulski JK et al.; The human CD1 proteins belong to a lipid-glycolipid antigen-presenting gene family and are related in structure and function to the MHC class I molecules . Previous mapping and DNA hybridization studies have shown that five linked genes located within a cluster on human chromosome 1q22-23 encode the CD1 protein family . We have analyzed the complete genomic sequence of the human CD1 gene cluster and found that the five active genes are distributed over 175,600 nucleotides and separated by four expanded intervening genomic regions (IGRs) ranging in length between 20 and 68 kb . The IGRs are composed mostly of retroelements including five full-length L1 PA sequences and various pseudogenes . Some L1 sequences have acted as receptors for other subtypes or families of retroelements . Alu molecular clocks that have evolved during primate history are found distributed within the HLA class I duplicated segments (duplicons) but not within the duplicons of CD1 . Phylogeny of the alpha3 domain of the class I-like superfamily of proteins shows that the CD1 cluster is well separated from HLA class I by a number of superfamily members including MIC (PERB11), HFE, Zn-alpha2-GP, FcRn, and MR1 . Phylogenetically, the human CD1 sequences are interspersed by CD1 sequences from other mammalian species, whereas the human HLA class I sequences cluster together and are separated from the other mammalian sequences . Genomic and phylogenetic analyses support the view that the human CD1 gene copies were duplicated prior to the evolution of primates and the bulk of the HLA class I genes found in humans . In contrast to the HLA class I genomic structure, the human CD1 duplicons are smaller in size, they lack Alu clocks, and they are interrupted by IGRs at least 4 to 14 times longer than the CD1 genes themselves . The IGRs seem to have been created as "buffer zones" to protect the CD1 genes from disruption by transposable elements.

Am J Physiol Heart Circ Physiol, 2001 Nov, 281(5), H2105 - 12
Chronic estrogen depletion alters adenosine diphosphate-induced pial arteriolar dilation in female rats; Xu HL et al.; We examined pial arteriolar reactivity to a partially endothelial nitric oxide synthase (eNOS)-dependent vasodilator ADP as a function of chronic estrogen status . The eNOS-dependent portion of the ADP response was ascertained by comparing ADP-induced pial arteriolar dilations before and after suffusion of a NOS inhibitor, N(omega)-nitro-L-arginine (L-NNA; 1 mM) in intact, ovariectomized (Ovx), and 17beta-estradiol (E2)-treated Ovx females . We also examined whether ovariectomy altered the participation of other factors in the ADP response . Those factors were the following: 1) the prostanoid indomethacin (Indo); 2) the Ca2+-dependent K+ (K(Ca)) channel, iberiotoxin (IbTX); 3) the ATP-regulated K+ (K(ATP)) channel glibenclamide (Glib); 4) the K(Ca)-regulating epoxygenase pathway miconazole (Mic); and 5) the adenosine receptor 8-sulfophenyltheophylline (8-SPT) . In intact females, the eNOS-dependent (L-NNA sensitive) portion of the ADP response represented approximately 50% of the total . The ADP response was retained in the Ovx rats but L-NNA sensitivity disappeared . On E2 replacement, the initial pattern was restored . ADP reactivity was unaffected by Indo, Glib, Mic, and 8-SPT . IbTX was associated with 50-80% reductions in the response to ADP in the intact group that was nonadditive with L-NNA, and 60-100% reductions in the Ovx group . The present findings suggest that estrogen influences the mechanisms responsible for ADP-induced vasodilation . The continued sensitivity to IbTX in Ovx rats, despite the loss of a NO contribution, is suggestive of a conversion to a hyperpolarizing factor dependency in the absence of E2.

Exp Mol Med, 2001 Sep 30, 33(3), 136 - 44
Alterations of HLA class I and II antigen expression in preinvasive, invasive and metastatic cervical cancers; Ryu KS et al.; HLA expression is altered in a large variety of human cancers . We performed immunohistochemical staining on tissues from normal, preinvasive, invasive and metastatic cervical cancer tissues using anti-HLA class I or class II antibody . In tissues from normal squamous epithelium, carcinoma in situ (CIS) and microinvasive carcinoma (MIC), the expressions of HLA-B, C heavy chains and class II heavy chain were significantly decreased as disease progressed . When the expression patterns were compared between primary and metastatic squamous cell carcinoma (SCC) lesions, statistically significant down-regulation of HLA class I and class II antigen in metastatic lesions was observed . The rates of HLA-B, C heavy chains and class II heavy chain expressions were all significantly down-regulated compared to the down-regulation rate of class I beta2-microglobulin (beta2m) in invasive squamous lesions, and the expressions of class II heavy chain in metastatic lesions was decreased further than that in primary lesions . Unlike SCC, the degree of HLA class I and class II loss was not evident as disease progressed in early stage of adenocarcinoma . In invasive adenocarcinoma lesions, only the expression of HLA-B, C heavy chains was decreased and no differences were seen in HLA-B, C heavy chain expression patterns between primary and metastatic lesions . These results suggest that alterations of HLA class I and II expressions seem to occur at a particular step in cervical cancer development and depend on tissue types: when the tumor becomes invasive and starts to metastasize.

Farmaco, 2001 Aug, 56(8), 621 - 3
Antituberculosis agents II . Evaluation of in vitro antituberculosis activity and cytotoxicity of some 2-(1-methyl-5-nitro-2-imidazolyl)-1,3,4-thiadiazole derivatives; Foroumadi A et al.; Using the radiometric BACTEC 460-TB methodology, the minimum inhibitory concentration (MIC) of a series of 2-(1-methyl-5-nitro-2-imidazolyl-1,3,4-thiadiazole-5-alkylsulfides, alkylsulfoxides and alkylsulfones which had been reported previously as antifungal agents, were determined . Active compounds were also screened by serial dilution to assess toxicity to a VERO cell line . The results indicate that compounds bearing a primary alkylthio substitution displayed good antituberculosis activity (MIC = 3.13-6.25 microg/ml) . Oxidation to sulfone abolished the antituberculosis activity in methyl and propyl derivatives while the ethylsulfonyl analogue was active (MIC = 1.56 microg/ml) . The cytotoxic effects indicate that 2-(1-methyl-5-nitro-2-imidazolyl)-5-methylthio-1,3,4-thiadiazole was the least toxic compound (IC50 > 10 microg/ml) . Generally, all compounds showed a low selectivity index.

Chin Med J (Engl), 1999 Jun, 112(6), 484 - 8
In vitro activities of five antifungal agents against pathogenic Exophiala species; Li D et al.; OBJECTIVES: To examine the in vitro activities of five antifungal agents against common pathogenic Exophiala species; to detect the minimum inhibitory concentration (MIC) discrepancies among different drugs and different species; to evaluate the role of the influence factors in MIC determination; and to establish a standard method for the antifungal susceptibility testing of Exophiala spp . METHODS: MICs of itraconazole (ICZ), fluconazole, ketoconazole, amphotericin B (AmB) and 5-flucytosine against 5 species (57 strains) of Exophiala spp were determined by modified NCCLS M27-A broth microdilution method . Two inoculum sizes (0.5 x 10(3)-2.5 x 10(3) CFU/ml and 0.5 x 10(4)-2.5 x 10(4) CFU/ml) and three incubation times (4, 7 and 10 days) were evaluated . The minimum fungicidal concentrations (MFC) of itraconazole against 20 Exophiala strains were also detected . RESULTS: All the tested Exophiala species were sensitive to AmB and ICZ . MICs varied among different species and drugs . Resistant strains to different drugs existed in this genus . The proper inoculum density was about 0.5 x 10(3) CFU/ml to 2.5 x 10(3) CFU/ml, and the 4-day incubation time was suitable for more than 90% of the strains . CONCLUSIONS: AmB or ICZ may serve as the first choice in treatment of phaeohyphomycosis . A standard microdilution procedure for MIC detection of Exophiala species is established in this experiment . The inoculum size and incubation time are the essential factors in the standardization of antifungal susceptibility testing of filamentous fungi . We hope this experiment may correlate well with the clinical treatment of phaeohyphomycosis caused by Exophiala species.

Antimicrob Agents Chemother, 2001 Nov, 45(11), 3148 - 55
Pharmacokinetics of cefepime during continuous renal replacement therapy in critically ill patients; Malone RS et al.; The pharmacokinetics of cefepime were studied in 12 adult patients in intensive care units during continuous venovenous hemofiltration (CVVH) or continuous venovenous hemodiafiltration (CVVHDF) with a Multiflow60 AN69HF 0.60-m(2) polyacrylonitrile hollow-fiber membrane (Hospal Industrie, Meyzieu, France) . Patients (mean age, 52.0 +/- 13.0 years {standard deviation}; mean weight, 96.7 +/- 18.4 kg) received 1 or 2 g of cefepime every 12 or 24 h (total daily doses of 1 to 4 g/day) by intravenous infusion over 15 to 30 min . Pre- and postmembrane blood (serum) samples and corresponding ultrafiltrate or dialysate samples were collected 1, 2, 4, 8, and 12 or 24 h (depending on dosing interval) after completion of the drug infusion . Drug concentrations were measured using validated high-performance liquid chromatography methods . Mean systemic clearance (CL(S)) and elimination half-life (t(1/2)) of cefepime were 35.9 +/- 6.0 ml/min and 12.9 +/- 2.6 h during CVVH versus 46.8 +/- 12.4 ml/min and 8.6 +/- 1.4 h during CVVHDF, respectively . Cefepime clearance was substantially increased during both CVVH and CVVHDF, with membrane clearance representing 40 and 59% of CL(S), respectively . The results of this study confirm that continuous renal replacement therapy contributes substantially to total CL(S) of cefepime and that CVVHDF appears to remove cefepime more efficiently than CVVH . Cefepime doses of 2 g/day (either 2 g once daily or 1 g twice daily) appear to achieve concentrations adequate to treat most common gram-negative pathogens (MIC <or= 8 microg/ml) during CVVH or CVVHDF.

Antimicrob Agents Chemother, 2001 Nov, 45(11), 3029 - 36
Development and validation of limited-sampling strategies for predicting amoxicillin pharmacokinetic and pharmacodynamic parameters; Suarez-Kurtz G et al.; Amoxicillin plasma concentrations (n = 1,152) obtained from 48 healthy subjects in two bioequivalence studies were used to develop limited-sampling strategy (LSS) models for estimating the area under the concentration-time curve (AUC), the maximum concentration of drug in plasma (C(max)), and the time interval of concentration above MIC susceptibility breakpoints in plasma (T>MIC) . Each subject received 500-mg amoxicillin, as reference and test capsules or suspensions, and plasma concentrations were measured by a validated microbiological assay . Linear regression analysis and a "jack-knife" procedure revealed that three-point LSS models accurately estimated (R(2), 0.92; precision, <5.8%) the AUC from 0 h to infinity (AUC(0-infinity)) of amoxicillin for the four formulations tested . Validation tests indicated that a three-point LSS model (1, 2, and 5 h) developed for the reference capsule formulation predicts the following accurately (R(2), 0.94 to 0.99): (i) the individual AUC(0-infinity) for the test capsule formulation in the same subjects, (ii) the individual AUC(0-infinity) for both reference and test suspensions in 24 other subjects, and (iii) the average AUC(0-infinity) following single oral doses (250 to 1,000 mg) of various amoxicillin formulations in 11 previously published studies . A linear regression equation was derived, using the same sampling time points of the LSS model for the AUC(0-infinity), but using different coefficients and intercept, for estimating C(max) . Bioequivalence assessments based on LSS-derived AUC(0-infinity)'s and C(max)'s provided results similar to those obtained using the original values for these parameters . Finally, two-point LSS models (R(2) = 0.86 to 0.95) were developed for T>MICs of 0.25 or 2.0 microg/ml, which are representative of microorganisms susceptible and resistant to amoxicillin.

Hum Immunol, 2001 Oct, 62(10), 1153 - 8
Tumor necrosis factor A and MHC class I chain related gene A (MIC-A) polymorphisms in Swedish patients with cervical cancer; Ghaderi M et al.; Human papillomaviruses type 16 and 18 are the major cause of cervical cancer . However, genetic factors contribute to the propensity of persistent HPV infection and cervical carcinoma . Allelic variants of the human leukocyte genes have shown to be associated with cervical neoplasia . The strongest associations have been found with the genes in the HLA class II region . The aim of this study was to analyze the association of two non-HLA class II markers with invasive cervical cancer . Microsatellite polymorphism of the TNFA gene located in the class III region and a short tandem repeat polymorphism of the MICA gene located in the centromeric end of the HLA class I region were analyzed . Eighty-five patients and 120 matched control individuals from a population-based cohort from Northern Sweden participated in this nested case-control study . MICA was not associated with cervical carcinoma . TNFa-11 frequency was increased in the HPV18 DNA positive patients (OR = 2.84, p = 0.0481, CI = 1.04-7.78, pc = NS) . TNFa-11 was not associated with susceptibility to HPV16 infection, but it increased the risk for cervical cancer with the HLA DQ6 (DQA 1*0102-DQB 1*0602) haplotype . Our findings indicate that the association of TNFA with cervical cancer is different with CIN . The extended HLA DQ6-TNFa-11 haplotype is increasing the risk for development of cervical cancer significantly (OR = 3.08, p = 0.0104, CI = 1.30-7.31).

Yao Xue Xue Bao, 1997 Oct, 32(10), 731 - 4
{Regulation of NGF on intra-synaptosomal free {Ca2+}i in mouse brain}; Xu XH et al.; In the present study, the influence of direct intrahippocampal injection of NGF (nerve growth factor) on synaptosomal free {Ca2+}i in 1-month-old and 18-month-old mice was observed . The regulation of NGF on synaptosomal free {Ca2+}i in vitro was also investigated . The synaptosomal free {Ca2+}i in the main brain regions (cerebral cortex, hippocampus, cerebellum, diencephalon) of these mice was measured by fluorescent probe Fura-2 of Ca2+ indicator and an AR-CM-MIC cation measurement system . The results are as follows: (1) NGF showed no detectable effect on hippocampal intrasynaptosomal free {Ca2+}i in 1-month-old mice, but appropriate dose of NGF was shown to markedly reduce {Ca2+}i in 18-month-old mice . (2) Under the condition of low {Ca2+}i level caused by the drug, NGF was found to increase {Ca2+}i; conversely, was reduced {Ca2+}i when intracellular Ca2+ was overloaded . These results suggest that NGF has dual regulation on Ca2+ level in brain and this might be an important mechanism for its action in improving memory lesion, in aged person.

Yao Xue Xue Bao, 1997 Oct, 32(10), 721 - 5
{Effect of berberine on cytosolic free calcium of rat myocardial cells in vitro}; Li XT et al.; In order to verify the inotropic effect of berberine (Ber) on myocardial tissues, the effect of Ber on cytosolic free calcium {Ca2+}i in cultured rat single myocardial cells were examined by using Fura-2/AM and AR-CM-MIC cation measurement system . Ber (10-400 mumol.L-1) was found to increase the resting {Ca2+}i and saturate at about 200 mumol.L-1 . The {Ca2+}i increase was not depressed by atropine, phentolamine, propranolol and tetrodotoxin, but was inhibited by verapamil (Ver) 3 mumol.L-1 or CoCl2 10 mmol.L-1 . The {Ca2+}i increase induced by Ber (30 mumol.L-1) added to stimulating agents as KCl, CaCl2, NE or Oua was stronger than that induced by stimulating agents only and Ver 3 mumol.L-1 still showed inhibitory effects on the {Ca2+}i increase . Under Ca(2+)-free or Ca(2+)-, K+, Na(+)-free conditions, Ber (30-200 mumol.L-1) still showed ascending effects on {Ca2+}i, and Ver 3 mumol.L-1 only inhibited the Ber effects in Ca(2+)-free condition . The results suggest that Ber might induce moderate {Ca2+}i increase via Ca2+ influx and intracellular Ca2+ release, and show positive inotropic effect.

Yao Xue Xue Bao, 1997 Nov, 32(11), 819 - 23
{8-(N,N-diethylamino)-n-octyl-3,4,5-trimethoxybenzoate(TMB-8) reduced the elevation of {Ca2+}i induced by BHQ, NE and KCl in cultured single smooth muscle cells of the calf basilar artery}; Wang B et al.; The effect of 8-(N, N-diethylamino)-n-octyl-3,4,5-trimethoxybenzoate (TMB-8) on the elevation of {Ca2+}i induced by 2, 5-di (tert-butyl)-1, 4-benzohydroquinone (BHQ), norepinephrine (NE), KCl in cultured single smooth muscle cells of the calf basilar artery was studied by a system of measurement of AR-CM-MIC, using Fura-2/AM as a fluoresent indicator . In the presence of extracellular Ca2+ 1.3 mmol.L-1, the resting {Ca2+}i was not changed by TMB-8 (10, 30 and 100 mumol.L-1), but the elevation of {Ca2+}i induced by BHQ, NE and KCl were reduced by TMB-8 (30 mumol.L-1) significantly . In Ca2+ free Hank's solution containing EGTA 0.1 mmol.L-1, the resting {Ca2+}i was markedly reduced by TMB-8 (10, 30 and 100 mumol.L-1), and the increase of {Ca2+}i evoked by BHQ and NE was blocked completely by TMB-8 (30 mumol.L-1) . The result suggested that TMB-8 inhibited the Ca2+ release from intracellular stores or increased the up-take of Ca2+ into sarcoplasmic reticulum and the inhibition of Ca(2+)-influx from extracellular site may be an indirect machanism.

J Chemother, 2001 Aug, 13(4), 395 - 401
Temporary interruption of ceftazidime continuous infusion without reduction of activity: a computer-assisted simulation; Castagnola E et al.; Continuous infusion (CI) of ceftazidime has been demonstrated to add clinical advantages in the treatment of infection of neutropenic cancer patients, especially in the presence of gram-negative bacteremia . However, this particular administration route is not always feasible in this particular clinical setting because of the patient's need of additional care or drug administration . The aim of this study was to evaluate, through a computer-assisted simulation, the modifications of drug concentration in presence of a single or repeated 2-hour interruptions of CI ceftazidime in critically ill patients . Our analysis shows that a loading dose of 20 mg/kg, followed by a CI of 100/mg/kg/die, should be able to maintain efficacious plasma concentrations in all subjects, even when it is interrupted for a 2-hour period every 8 hours . Plasma concentrations after interruption should not fall below 8 microg/mL and for about 65-80% of time should reach levels equal to 5 times the MIC of the infecting pathogen . A 2-hour interruption of CI ceftazidime up to 3 times a day is likely to represent a safe and efficacious administration regimen that may enhance the management of the treatment of infectious complications in critically ill patients such as neutropenic cancer patients.

Perit Dial Int, 2001 Jul-Aug, 21(4), 378 - 85
Intravenous vancomycin pharmacokinetics in automated peritoneal dialysis patients; Manley HJ et al.; The pharmacokinetics of intravenous (i.v.) vancomycin was studied in automated peritoneal dialysis (APD) patients who received a single i.v . dose of vancomycin (15 mg/kg total body weight) . Dialysate samples were collected at the beginning, middle, and end of dwells 1-3 (on-cycler), and at the end of dwells 4 and 5 (off-cycler), for a 24-hour period . Blood samples were collected at the beginning, middle, and end of dwells 1-3 (on-cycler), and at the end of dwell 5 (off-cycler) for a 24-hr period . Pharmacokinetics parameters were calculated assuming a one-compartment model . Glomerular filtration rate (GFR) and vancomycin clearance (CI) values were normalized to 1.73 m2 . Ten patients {4 males, 6 females; 47.4 +/- 9.9 years of age (mean +/- SD)} who had received PD for a median 3.5 months (range 2-66 months) were studied . Dwell times were 2.3 +/- 0.1 hours on cycler and 7.3 +/- 0.1 hours off cycler . Vancomycin half-life was significantly different on-cycler than off-cycler (11.6 +/- 5.2 hr vs 62.8 +/- 33.0 hr; p < 0.001) . Vancomycin total CI (CI(T)) was 7.4 +/- 2.0 mL/min . Renal CI (CI(R)) and PD CI (CI(PD)) accounted for 23.6% and 28.0% of CI(T) . respectively . CI(R) correlated with GFR (CI(R) = 0.90 GFR - 1.01; r2 = 0.79; p = 0.008) . Mean vancomycin serum and dialysate end-of-dwell concentrations were above minimum inhibitory concentration of susceptible organisms (5 micro/mL) for the first cycler and the second ambulatory exchanges only . The results of this study suggest that, to provide adequate concentrations for susceptible organisms over a 24-hour period, current intermittent vancomycin dosing recommendations for PD-related peritonitis need to be changed to 35 mg/kg intraperitoneally on day 1, then 15 mg/kg i.p . thereafter (i.e., once daily) in APD patients.

Cancer Res, 2001 Oct 1, 61(19), 6996 - 7001
Secreted and cell surface genes expressed in benign and malignant colorectal tumors; Buckhaults P et al.; Serial analysis of gene expression was used to identify transcripts encoding secreted or cell surface proteins that were expressed in benign and malignant tumors of the colorectum . A total of 290,394 tags were analyzed from normal, adenomatous, and cancerous colonic epithelium . Of the 21,343 different transcripts observed, 957 were found to be differentially expressed between normal tissue and adenoma or between normal tissue and cancer . Forty-nine transcripts were elevated > or =20-fold in adenomas, 40 transcripts were elevated > or =20-fold in cancers, and 9 transcripts were elevated > or =20-fold in both . Products of six of these nine transcripts (TGFBI, LYS, RDP, MIC-1, REGA, and DEHL) were predicted to be secreted or to reside on the cell surface, and these were analyzed in more detail . The abnormal expression levels predicted by serial analysis of gene expression were confirmed by quantitative PCR analyses of each of these six genes . Moreover, the cell types responsible for the elevated expression were identified by in situ hybridization and by PCR analyses of epithelial cells immunoaffinity purified from primary tumors . This study extends knowledge of the differences in gene expression that underlie various stages of neoplasia and suggests specific diagnostic approaches that may be useful for the early detection of colorectal neoplasia.

J Ethnopharmacol, 2001 Nov, 78(1), 59 - 66
Anti-amoebic activity of plant compounds from Virgilia oroboides and Chlorophora excelsa; Padayachee T et al.; The anti-amoebic activity of four plant extracts: maackiain and formononetin from Virgilia oroboides and chlorophorin and Iroko from Chlorophora excelsa, were evaluated . Anti-protozoal tests conducted on trophozoites of Entamoeba histolytica established that all four compounds had an affect on the trophozoites to some degree . Chlorophorin showed the highest anti-protozoal activity with an MIC of 0.25 microg/ml followed by maackiain and Iroko with MICs of 1 microg/ml . Chlorophorin and Iroko induced the release of acid phosphatase . Chlorophorin reduced alpha amylase levels by 89% . Formononetin and maackiain had a minimal effect on the enzyme levels . Ultrastructural changes occurred in trophozoites treated with plant compounds . The degree of destruction of the trophozoites increased with an increase in compound concentration . Trophozoite destruction was initiated by the disintegration of the nucleus and culminated with the rupture of the cytoplasmic membrane . Maackiain was the only compound that showed some level of mutagenicity . Formononetin and Iroko were very slightly mutagenic, while chlorophorin was non-mutagenic . In addition, none of the compounds tested showed cytopathic effects on any of the cell lines tested . Chlorophorin and Iroko exhibit the potential to be exploited as natural multi-functional safe control agents in the treatment of bacterial, fungal and protozoal infections.

J Antimicrob Chemother, 2001 Oct, 48(4), 557 - 61
Evaluation of daptomycin susceptibility testing by Etest and the effect of different batches of media; Fuchs PC et al.; One hundred and ninety-five Gram-positive bacteria representing 17 species were tested for susceptibility to daptomycin by broth microdilution and Etest methods . The geometric mean daptomycin MIC was 0.46 mg/L by broth microdilution tests and 0.73 mg/L by Etest . The concentration of calcium in 12 different batches of agar varied from 4 to 36 mg/L . Daptomycin Etest MICs varied inversely with the calcium concentration . Etest daptomycin MICs for quality control strains were within proposed quality control range on media with >20 mg/L of calcium . Monitoring the calcium levels of agar media by testing appropriate quality control strains is important for daptomycin Etests.

J Comp Neurol, 2001 Oct 8, 439(1), 32 - 45
Expression of growth differentiation factor-15/ macrophage inhibitory cytokine-1 (GDF-15/MIC-1) in the perinatal, adult, and injured rat brain; Schober A et al.; We and others have recently cloned a new member of the transforming growth factor-beta superfamily, growth differentiation factor-15/ macrophage inhibitory cytokine-1 (GDF-15/MIC-1) . Using in situ hybridization and immunohistochemistry, we determined the distribution of GDF-15/MIC-1 mRNA and protein in the perinatal and cryolesioned adult rat brain . The choroid plexus epithelium of all ventricles represents the site of strongest and almost exclusive mRNA expression in the normal perinatal and adult brain . The newborn rat brain reveals GDF-15/MIC-1 immunoreactivity (ir) in ependymal cells lining the ventricles, in the striatal subventricular zone, and in populations of nonneural cells of the thalamic/hippocampal lamina affixa, in addition to that in the choroid plexus . Unilateral cryogenic cortical lesioning induced a significant increase of GDF-15/MIC-1 mRNA expression and ir at the lesion site and expression in presumed neurons within the dorsal thalamic area . At the lesion site, GDF-15/MIC-1-producing cells showed immuncytochemical features of neurons, macrophages, and activated microglial cells . Fluorescent microscopy revealed both intra- and extracellular GDF-15/MIC-1 ir . Up-regulation of GDF-15/MIC-1 in activated macrophages (Mstraight phi) is also supported by RT-PCR, ICC, and Western blot experiments showing pronounced induction of GDF-15/MIC-1 expression (mRNA and protein) in retinoic acid/phorbol ester-stimulated human M phi . Our data suggest that 1) GDF-15/MIC-1 is secreted into the cerebrospinal fluid and 2) in the newborn brain may penetrate through the ependymal lining and act on developing neurons and/or glial cells . As a constituent of cells in the lamina affixa, the protein might be involved in the regulation of mesenchyme-epithelial interactions . Finally, GDF-15/MIC-1 may also act within the antiinflammatory cytokine network activated in CNS lesions .

Microbiology, 2001 Oct, 147(Pt 10), 2769 - 81
Polycations increase the permeability of Mycobacterium vaccae cell envelopes to hydrophobic compounds; Korycka-Machala M et al.; Polycations {protamine, polymyxin B nonapeptide (PMBN) and polyethyleneimine (PEI)} have been shown to increase the cell wall permeability of Mycobacterium vaccae to highly hydrophobic compounds, as manifested in enhanced intracellular bioconversion of beta-sitosterol to 4-androsten-3,17-dione (AD) and 1,4-androstadien-3,17-dione (ADD), and cell sensitization to erythromycin and rifampicin . The quantity of AD(D) formed per biomass unit was twice as high in the presence of PMBN and PEI, and three times higher with protamine . The sensitization factor, i.e . the MIC(50) ratio of the control bacteria to those exposed to polycations, ranged from 4 to 16, depending on the polycation/antibiotic combination . Non-covalently bound free lipids were extracted from the control and polycation-treated cells and fractionated with the use of chloroform, acetone and methanol . Chloroform- and acetone-eluted fractions (mainly neutral lipids and glycolipids, respectively) showed significant polycation-induced alterations in their quantitative and qualitative composition . The fatty acid profile of neutral lipids was reduced in comparison to control, whereas acetone-derived lipids were characterized by a much higher level of octadecenoic acid (C(18:1)) and a considerably lower content of docosanoic acid (C(22:0)), the marker compound of mycolate-containing glycolipids . Methanol-eluted fractions remained unaltered . Cell-wall-linked mycolates obtained from delipidated cells were apparently unaffected by the action of polycations, as judged from the TLC pattern of mycolic acid subclasses, the mean weight of mycolate preparations and the C(22:0) acid content in the mycolates, determined by GC/MS and pyrolysis GC . The results suggest the involvement of the components of non-covalently bound lipids in the outer layer in the M . vaccae permeability barrier.

Int J Antimicrob Agents, 2001, 18 Suppl 1, S17 - 23
Macrolides: pharmacokinetics and pharmacodynamics; Van Bambeke F et al.; Three pharmacokinetic/pharmacodynamic parameters--(i) the peak concentration to the minimum inhibitory concentration ratio (C(max)/MIC); (ii) the area under the concentration-time curve to MIC ratio (AUC(24h)/MIC); and (iii) the time the concentration exceeds the MIC (T>MIC)--are important predictors of the clinical efficacy of antibiotics . For antibiotics with pronounced concentration-dependent killing, such as the fluoroquinolones or the aminoglycosides, C(max)/MIC and AUC(24)/MIC are the main factors that establish efficacy . Antibiotics with a weak, or no, concentration dependency generally have their efficacy linked to T>MIC, and these include the beta-lactams and the conventional macrolides . Antibiotics with weak concentration-dependent effects, but with prolonged persistent effects, such as tetracyclines and azithromycin, have their activity mostly related to the AUC(24)/MIC . By applying these concepts to current antibiotics, and also to the development of novel agents, it is possible to optimise their dosages and administration schedules . This will maximise therapeutic efficacy, may prevent or delay the emergence of bacterial resistance to antibiotics, and can certainly minimise side-effects.

J Gastroenterol, 2000 Dec, 35(12), 933 - 40
Ewing's sarcoma/peripheral primitive neuroectodermal tumor (pPNET) arising in the omentum as a multilocular cyst with intracystic hemorrhage; Tanida S et al.; A rare case of Ewing's sarcoma/peripheral primitive neuroectodermal tumor arising in the greater omentum in a 41-year-old man is reported . The patient presented with a hemorrhagic mesenteric cyst that was disclosed by the results of an abdominal echogram, a computed tomography scan, and magnetic resonance imaging . A laparotomy showed a multilocular cyst with intra-cystic hemorrhage . Histologically, the tumor wall consisted of sheets of small round cells separated by thick desmoplastic stroma . Rosette formations or ribbon-like cell arrangements were absent . Further pathological examination revealed that the membrane of the tumor cells was positive for MIC-2, and negative for epithelial membrane antigen, cytokeratin, and desmin, which are usually positive in intra-abdominal desmoplastic small round-cell tumors . An EWS/FLI1 fused transcript was detected by reverse transcription-polymerase chain reaction . These findings confirmed the diagnosis of Ewing's sarcoma/peripheral primitive neuroectodermal tumor . The patient died of tumor recurrence 4 months after his first admission . The autopsied tumor tissue exhibited neural differentiation in certain regions . To our knowledge, this is the first case to be reported of Ewing's sarcoma/peripheral primitive neuroectodermal tumor arising in the omentum with unique pathological features and the occurrence of partial neural differentiation during the clinical course . This case pointed out to us, as gastroenterologists, that only thorough examination confirms a definitive diagnosis of small round-cell tumor of the abdomen, it also shows that Ewing's sarcoma/peripheral primitive neuroectodermal tumor should be included in the differential diagnosis of cystic lesions in the omentum.

Drug Metab Dispos, 2001 Oct, 29(10), 1332 - 6
Influence of microsomal concentration on apparent intrinsic clearance: implications for scaling in vitro data; Kalvass JC et al.; The influence of microsomal concentration on unbound fraction (fu(mic)), half-life (t(1/2)), apparent intrinsic clearance (CL(int,app)) and apparent Michaelis-Menten constant (K(m,app)) was examined for two compounds, one representative of high nonspecific binding to microsomes (compound A) and one representative of low (compound B) . Kinetic parameters were estimated for the two probe compounds at two human microsomal protein concentrations (0.46 and 2.3 mg/ml) and cytochrome P450 concentrations (0.20 and 1.0 microM), representing a 5-fold difference in microsomal concentration . For compound A, fu(mic) and CL(int,app) were inversely proportional to microsomal concentration . Conversely, the K(m,app) of compound A was proportional to microsomal concentration and the half-life was unchanged . For compound B, half-life was inversely proportional to microsomal concentration . In this case, fu(mic), CL(int,app), and K(m,app) were not proportionally influenced . The experimental observations were entirely consistent with that predicted by a mathematical relationship between microsomal concentration, fu(mic), t(1/2), CL(int,app), and K(m,app) . These results demonstrate that when nonspecific binding is extensive, CL(int,app) is dependent on the arbitrary choice of microsomal concentration included in the incubation.

Lett Appl Microbiol, 2001 Oct, 33(4), 302 - 6
RAPD profile and antibiotic susceptibility of Xylella fastidiosa, causal agent of citrus variegated chlorosis; Lacava PT et al.; AIMS: The aim of this study was to evaluate the diversity of Xylella fastidiosa isolated from citrus trees affected by Citrus Variegated Chlorosis (CVC) . METHODS AND RESULTS: The antibiotic susceptibility by agar disc diffusion and minimum inhibitory concentration (MIC) methods was observed for all drug evaluated, except for penicillin-G . Genetic diversity by RAPD analysis revealed three major groups (citrus, coffee and grapevine), being the citrus group more similar with the coffee group than with the grapevine group . CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: This study highlights the possibility to use these antibiotics susceptibility as markers in the development of a cloning vector and penicillin-G could be used as a selective marker for the isolation of X . fastidiosa from citrus plants.

Clin Ther, 2001 Aug, 23(8), 1231 - 44
Pharmacokinetics and pharmacodynamics of aztreonam and tobramycin in hospitalized patients; Smith PF et al.; BACKGROUND: Pharmacokinetic/pharmacodynamic (PK/PD) optimization of antibiotic therapy has been shown to improve outcomes in several antibiotic classes . Despite the frequent use of beta-lactams, clinical data in humans remain limited . OBJECTIVE: This study evaluated the relationship between serum pharmacokinetics, pharmacodynamics, pathogen susceptibility, and clinical outcomes in patients receiving aztreonam or tobramycin monotherapy . METHODS: The case-report forms of hospitalized patients who received either aztreonam or tobramycin for a bacterial infection in 3 clinical trials conducted between 1982 and 1984 were reviewed for the present study . A pathogen was identified for all included patients, and susceptibility testing was performed to determine the minimum inhibitory concentration (MIC) for each agent . Pharmacokinetic parameters for each antibiotic were determined using population modeling, and variables potentially related to outcomes were evaluated using tree-based modeling, logistic regression, and nonlinear regression methods . RESULTS: Data from 91 patients were analyzed, 68 treated with aztreonam monotherapy and 23 treated with tobramycin monotherapy . Of the types of infections treated, 39 were intra-abdominal, 42 involved the lower respiratory tract, and 10 involved the skin and skin structures . The pharmacodynamic ratio of the 24-hour area under the curve (AUC24) to the MIC was associated with clinical outcome for both antibiotics: aztreonam and to-bramycin patients with ratios meeting or exceeding the respective 24-hour inverse serum inhibitory titer breakpoints of 184 and 110 were significantly more likely to achieve a successful outcome than were those with ratios not meeting these values (P < 0.01) . The probabilities of clinical success in patients at or above and below the AUC24/MIC breakpoints were a respective 85% and 53% for aztreonam and 80% and 47% for tobramycin (both, P < 0.01) . When all patients were considered, the likelihood of achieving cure was 5.1 times greater in patients exceeding the target ratios (P < 0.01) . CONCLUSION: PK/PD optimization of both aztreonam and tobramycin is associated with improved patient outcomes.

Rev Inst Med Trop Sao Paulo, 2001 Jul-Aug, 43(4), 221 - 6
In vitro evaluation of quinidine sensitivity in Brazilian Plasmodium falciparum isolates: comparative analysis to quinine and chloroquine; Menezes CM et al.; Falciparum malaria represents a serious and an increasing world public health problem due to the acquired parasite's resistance to the most available drugs . In some endemic areas, quinidine, a diastereoisomer of the antimalarial quinine, has been employed for replacing the latter . In order to evaluate the use of quinidine as an alternative to the increasing loss of quinine effectiveness in Brazilian P . falciparum strains, as has been observed in the Amazon area, we have assayed quinidine, quinine and chloroquine . The in vitro microtechnique was employed . All isolates showed to be highly resistant to chloroquine . Resistance to quinine was not noted although high MIC (minimal inhibitory concentration) values have been observed . These data corroborate the decreasing sensitivity to quinine in strains from Brazil . Quinidine showed IC50 from 0.053 to 4.577 micromol/L of blood while IC50 from 0.053 to 8.132 micromol/L of blood was estimated for quinine . Moreover, clearance of the parasitemia was observed in concentrations lower than that used for quinidine in antiarrhythmic therapy, confirming our previous data . The results were similar to African isolate.

Antimicrob Agents Chemother, 2001 Oct, 45(10), 2891 - 6
Effects of AIDS and gender on steady-state plasma and intrapulmonary ethambutol concentrations; Conte JE Jr et al.; Our objective was to study the steady-state plasma and intrapulmonary orally administered ethambutol concentrations in healthy volunteers and subjects with AIDS . Ethambutol (15 mg/kg of body weight) was administered orally once daily to 10 men with AIDS, 10 healthy men, 10 women with AIDS, and 10 healthy women . The mean (+/-standard deviation {SD}) CD4 cell count for the 20 subjects with AIDS was (350 +/- 169) x 10(6) cells per liter . Blood was obtained for drug assay 2 h after the last dose and at the completion of bronchoalveolar lavage, performed 4 h after the last dose . Standardized bronchoscopy was performed without systemic sedation . The volume of epithelial lining fluid (ELF) was calculated by the urea dilution method . The total number of alveolar cells (AC) was counted in a hemocytometer, and differential cell counting was performed after cytocentrifugation . Ethambutol was measured by a new, sensitive and specific liquid chromotography-mass spectrometry method . The presence of AIDS, as defined in this study, or gender was without significant effect on the concentrations of ethambutol in plasma at 2 or 4 h or in ELF at 4 h following the last dose . Plasma drug concentrations (mean +/- SD) at 2 and 4 h were 2.1 +/- 1.2 and 2.1 +/- 0.8 microg/ml, respectively, and both values were not significantly different from the concentration of ethambutol in ELF at 4 h (2.2 +/- 1.1 microg/ml) . The concentration of ethambutol was significantly greater in AC in all four groups (range, 44.5 +/- 15.6 to 82.0 +/- 39.4 microg/ml) than in ELF or plasma and was approximately 30 to 240 times the reported MIC for ethambutol-susceptible strains of Mycobacterium tuberculosis . The AC ethambutol concentration (mean +/- SD) in the smoking women (97.2 +/- 32.1 microg/ml) was more than twice the concentration in all other nonsmoking subjects (45.2 +/- 16.8 microg/ml) combined (P < 0.05) . Two- and 4-h concentrations of ethambutol in plasma were not affected by AIDS status or gender . The high AC/plasma and AC/ELF concentration ratios suggest that substantial antimycobacterial activity resides in these cells . The data confirm earlier observations of active transport ex vivo of ethambutol into pulmonary macrophages.

Antimicrob Agents Chemother, 2001 Oct, 45(10), 2845 - 55
Pharmacokinetic and pharmacodynamic modeling of anidulafungin (LY303366): reappraisal of its efficacy in neutropenic animal models of opportunistic mycoses using optimal plasma sampling; Groll AH et al.; The compartmental pharmacokinetics of anidulafungin (VER-002; formerly LY303366) in plasma were characterized with normal rabbits, and the relationships between drug concentrations and antifungal efficacy were assessed in clinically applicable infection models in persistently neutropenic animals . At intravenous dosages ranging from 0.1 to 20 mg/kg of body weight, anidulafungin demonstrated linear plasma pharmacokinetics that fitted best to a three-compartment open pharmacokinetic model . Following administration over 7 days, the mean (+/- standard error of the mean) peak plasma concentration (C(max)) increased from 0.46 +/- 0.02 microg/ml at 0.1 mg/kg to 63.02 +/- 2.93 microg/ml at 20 mg/kg, and the mean area under the concentration-time curve from 0 h to infinity (AUC(0-infinity)) rose from 0.71 +/- 0.04 to 208.80 +/- 24.21 microg . h/ml . The mean apparent volume of distribution at steady state (V(ss)) ranged from 0.953 +/- 0.05 to 1.636 +/- 0.22 liter/kg (nonsignificant {NS}), and clearance ranged from 0.107 +/- 0.01 to 0.149 +/- 0.00 liter/kg/h (NS) . Except for a significant prolongation of the terminal half-life and a trend toward an increased V(ss) at the higher end of the dosage range after multiple doses, no significant differences in pharmacokinetic parameters were noted in comparison to single-dose administration . Concentrations in tissue at trough after multiple dosing (0.1 to 10 mg/kg/day) were highest in lung and liver (0.85 +/- 0.16 to 32.64 +/- 2.03 and 0.32 +/- 0.05 to 43.76 +/- 1.62 microg/g, respectively), followed by spleen and kidney (0.24 +/- 0.65 to 21.74 +/- 1.86 and <0.20 to 16.92 +/- 0.56, respectively) . Measurable concentrations in brain tissue were found at dosages of > or =0.5 mg/kg (0.24 +/- 0.02 to 3.90 +/- 0.25) . Implementation of optimal plasma sampling in persistently neutropenic rabbit infection models of disseminated candidiasis and pulmonary aspergillosis based on the Bayesian approach and model parameters from normal animals as priors revealed a significantly slower clearance (P < 0.05 for all dosage groups) with a trend toward higher AUC(0-24) values, higher plasma concentrations at the end of the dosing interval, and a smaller volume of distribution (P < 0.05 to 0.193 for the various comparisons among dosage groups) . Pharmacodynamic modeling using the residual fungal tissue burden in the main target sites as the primary endpoint and C(max), AUC(0-24), time during the dosing interval of 24 h with plasma drug concentration equaling or exceeding the MIC or the minimum fungicidal concentration for the isolate, and tissue concentrations as pharmacodynamic parameters showed predictable pharmacokinetic-pharmacodynamic relationships in experimental disseminated candidiasis that fitted well with an inhibitory sigmoid maximum effect pharmacodynamic model (r(2), 0.492 to 0.819) . However, no concentration-effect relationships were observed in experimental pulmonary aspergillosis using the residual fungal burden in lung tissue and survival as parameters of antifungal efficacy . Implementation of optimal plasma sampling in discriminative animal models of invasive fungal infections and pharmacodynamic modeling is a novel approach that holds promise of improving and accelerating our understanding of the action of antifungal compounds in vivo.

Antimicrob Agents Chemother, 2001 Oct, 45(10), 2781 - 6
Differential antifungal activity of isomeric forms of nystatin; Ostrosky-Zeichner L et al.; When nystatin is placed in RPMI and other biological fluids, there is loss of pure nystatin, with the development of two distinguishable chromatographic peaks, 1 and 2 . Peak 1 appears identical to commercially prepared nystatin . By nuclear magnetic resonance (NMR) and mass spectral analysis, peak 2 appears to be an isomer of peak 1 . The isomers are quantitatively and fully interconvertible . Formation of peak 2 is accelerated at a pH of >7.0 and ultimately reaches a near 55:45 (peak 1/peak 2 ratio) mixture . We sought to determine the relative activities of peaks 1 and 2 against Candida spp . Peak 2 consistently showed higher MICs when it was the predominant form during the experiment . Time-kill analyses showed that peak 2 required > or =8 x the concentration of peak 1 to produce a modest and delayed killing effect, which was never of the same magnitude as that produced by peak 1 . In both types of assays, the activity of peak 2 corresponded with intra-assay formation of peak 1 . Both MIC measurements and time-kill analysis suggest that peak 2 has considerably less activity, if any at all, against Candida spp . Peak 2 may serve as a reservoir for peak 1.

Med Mycol, 2001 Aug, 39(4), 369 - 71
Sporothrix schenckii sensitivity to voriconazole, itraconazole and amphotericin B; McGinnis MR et al.; One hundred clinical isolates of Sporothrix schenckii were tested against voriconazole, itraconazole and amphotericin B using a modification of the NCCLS M27-A in vitro yeast susceptibility testing procedure . NCCLS M38-P for moulds was not used because yeast forms may have been present when the test isolates were incubated at 35 +/- 1 degrees C . The minimum inhibitory concentration (MIC) values were: voriconazole 0.5-8 (geometric mean titer 6.50) microg ml(-1) ; itraconazole 0.03-8 (geometric mean titer 1.56) microg ml(-1); and amphotericin B 0.25-2 (geometric mean titer 1.23) microg ml(-1) . The minimum fungicidal concentration (MFC) values were: voriconazole 2-8 (geometric mean titer 7.67) microg ml(-1); itraconazole 0.125-8 (geometric mean titer 7.41) microg ml(-1); and amphotericin B 0.125-2 (geometric mean titer 1.53) microg ml(-1) . Based upon MIC values, sensitivity to amphotericin B is strain-dependent . S . schenckii is more sensitive to itraconazole than voriconazole based upon a comparison of MIC geometric means, even though the MIC ranges were essentially the same.

Appl Immunohistochem Mol Morphol, 2001 Sep, 9(3), 255 - 60
Immunohistochemical detection of EWS and FLI-1 proteinss in Ewing sarcoma and primitive neuroectodermal tumors: comparative analysis with CD99 (MIC-2) expression; Llombart-Bosch A et al.; The molecular analysis of the t(11;22) rearrangement involving EWS/FLI-1 genes is likely to be of diagnostic value in Ewing sarcoma (ES) and primitive neuroectodermal tumors (PNET) . The objective of the current study was to analyze the immunohistochemical expression of the EWS and FLI-1 proteins in a group of small round-cell tumors (SRCT) to determine their specificity and relevance in their differential diagnosis . Forty-eight cases-10 conventional ES, 4 large-cell ES, 5 PNET, 9 neuroblastomas (NB), 6 undifferentiated synovial sarcomas (SS), 5 rhabdomyosarcomas (RB), 5 non-Hodgkin lymphomas (NHL), 1 round-cell liposarcoma, and 3 mesenchymal chondrosarcomas-were analyzed . Immunocytochemistry was performed on paraffin sections after the LSAB method and antigen retrieval using ethylenediaminetetraacetic acid buffer (pH 6) . Primary antibodies included FLI-1 (C-19), EWS (N-18), EWS (C-19), and CD99 (MIC-2) . As expected, CD-99 expression was found in 100% of ES/PNET cases, in 2 cases of RB, 2 SS, and 1 NHL . FLI-1 protein was observed as nuclear staining in 16 cases of ES/PNET (84%) and in 4 cases of NHL, 2 NB, and 3 SS . Normal endothelial cells and lymphocytes also were positive . EWS expression (both proteins N-18 and C-19) was detected not only in 95% of ES/PNET cases but also in more than 50% of cases from the other tumoral types (4 of 9 and 7 of 9 NB, 5 of 6 and 6 of 6 SS, 3 of 5 and 5 of 5 RB, and 2 of 5 and 3 of 5 NHL, respectively) . Whereas EWS expression does not appear specific for ES/PNET, analysis of FLI-1 expression together with CD-99 is a powerful marker for ES/PNET and important factors in the differential diagnosis of SRCT.

Arch Pharm Res, 2001 Aug, 24(4), 297 - 9
In vitro anti-Helicobacter pylori activity of panaxytriol isolated from ginseng; Bae EA et al.; This study investigated the effect that some polyacetylenes and protopanaxatriol, which were isolated from heated ginseng (family Araliaceae), have on inhibiting Helicobacter pylori (HP) growth . Among the compounds tested, panaxytriol was quite effective in inhibiting HP growth with an MIC of 50 microg/ml . Ginsenoside Rh1 and protopanaxatriol weakly inhibited H+/K+-ATPase from a rat stomach.

Pharmazie, 2001 Aug, 56(8), 610 - 2
Antituberculosis agents, I: Synthesis and antituberculosis activity of 2-aryl-1,3,4-thiadiazole derivatives; Foroumadi A et al.; The design, synthesis and antituberculosis activity of a series of 2-aryl-5-methylthio-1,3,4-thiadiazoles (5a-b), ethyl alpha-(5-aryl)-1,3,4-thiadiazole-2-ylthio)acetates (8a-b) and related compounds are described . All of the compounds were tested against Mycobacterium tuberculosis strain H37Rv in comparison to rifampicin . Six compounds exhibited a very good activity (MIC < 6.25 micrograms/ml, % Inhibition = 100).

Gynecol Oncol, 2001 Aug, 82(2), 360 - 3
Stamm gastrostomy for postoperative gastric decompression in gynecologic oncology patients; Hoffman MS et al.; OBJECTIVE: The aim of this study was to report our experience with Stamm gastrostomy for postoperative gastric decompression on a gynecologic oncology service . METHODS: This was an observational study over the 9-year period ending in October 2000 . A 24 French MIC gastrostomy with the Stamm technique was used . Criteria for a gastrostomy are outlined in the text and tubes placed for palliation were excluded . Patients were followed to determine length of open drainage, time to removal, related complications, and those who may have benefited . RESULTS: A total of 167 patients had 174 nonpalliative gastrostomy tubes placed . Nine percent of the patients had related complications with a major complication rate of 2% . Overall, approximately one-third of the patients appeared to benefit from the gastrostomy tube . CONCLUSION: The results support the safety and efficacy of the MIC gastric tube in patients undergoing extensive gynecologic cancer surgery which is likely to impact the function of the gastrointestinal tract .

Gynecol Oncol, 2001 Aug, 82(2), 350 - 4
High sensitivity of PCR in situ hybridization for the detection of human papillomavirus infection in uterine cervical neoplasias; Xiao Y et al.; OBJECTIVE: This study was designed in order to examine whether the sensitivity of PCR in situ hybridization (PISH) is superior to that of fluorescence in situ hybridization (FISH) and immunocytochemical staining (ICS) in detecting the presence of human papillomavirus (HPV) infection in uterine cervical neoplasias . METHODS: Three cell specimens were obtained from every 54 patients who had been histologically and cytologically diagnosed as severe dysplasia of the uterine cervix (SD), carcinoma in situ of the uterine cervix (CIS), or microinvasive carcinoma of the uterine cervix (MIC) . All of them were first stained by the Papanicolaou (pap) method before the experiments . After specimens were classified into SD (n = 17) and CIS/MIC groups (n = 37), the sensitivities of detection for HPV infection were compared among PISH, FISH, and ICS methods which had been carried out after decolorization of the pap stain . RESULTS: In the SD group, PISH demonstrated a 58.8% positive incidence of HPV, which was significantly higher than that (23.5%) shown by FISH (P < 0.05) . There was no significant difference in the positivities of HPV between the PISH and ICS (47.1%) methods . In the CIS/MIC group, PISH, FISH, and ICS showed 73.0, 43.2, and 54.1% positive incidences of HPV, respectively . There was significant difference in HPV positivities between PISH and FISH as well as ICS (P < 0.01, P < 0.05) . The total frequency of the detection of HPV infection by PISH was significantly higher than that by FISH (P < 0.01) and high with a marginal significance compared to that by ICS (P = 0.051) . Moreover, the morphologic change of a single cell and the HPV-positive signals could be observed simultaneously by the PISH technique . CONCLUSION: Because PISH possesses high sensitivity for the detection of HPV infection in morphologically intact cells from uterine cervical neoplasia, the PISH technique might have promising potential for application to the detection of HPV presence in situ in screening .

Cytogenet Cell Genet, 2001, 93(3-4), 265 - 9
Chicken microchromosomes are hypermethylated and can be identified by specific painting probes; Grutzner F et al.; Microdissection of single chicken microchromosomes (MICs) followed by degenerate oligonucleotide-primed (DOP) PCR allows the rapid generation of MIC-specific DNA libraries . Since some libraries derived from a single (or a few) chromosome(s) label the entire MIC fraction, the majority of chicken MICs share repetitive DNA sequences that are not found on the macrochromosomes . In evolutionarily distant bird species, MICs are invariably hypermethylated . Methylcytosine staining provides additional in situ evidence for the high gene content of MICs and strong compartmentalization of avian genomes .

J Clin Microbiol, 2001 Sep, 39(9), 3420 - 1
Peritoneal dialysis-associated peritonitis caused by Dermabacter hominis; Radtke A et al.; Dermabacter hominis was the cause of a peritoneal dialysis-associated peritonitis . D . hominis was identified by phenotypic criteria and by sequencing the 16S rRNA gene . Clinical cure was achieved with cefuroxime treatment despite the isolate's reduced susceptibility to this drug (MIC, 12 mg/liter) on in vitro testing . The successful treatment was probably due to the high concentrations attained by intraperitoneal administration of the drug.

Clin Infect Dis, 2001 Sep 15, 33 Suppl 3, S238 - 9
Role of pharmacokinetics and pharmacodynamics: does the dose matter?
MacGowan AP.
Antibiotic dose is important in determining serum area-under-the-curve (AUC) and peak serum concentration (C(max)), as well as the time the serum concentration remains over the pathogen minimum inhibitory concentration (T>MIC) . However, dose is not the sole determinant of these factors; they are modified by absorption, clearance, and frequency of dosing . It is difficult to relate dose to clinical outcome in humans, but pharmacodynamic parameters (AUC/MIC, C(max)/MIC, or T>MIC) have been related to clinical and bacteriological efficacy or emergence of resistance to aminoglycosides, fluoroquinolones, glycopeptides, and beta-lactams.

Clin Infect Dis, 2001 Sep 15, 33 Suppl 3, S233 - 7
Does the dose matter?
Craig WA.
Pharmacokinetic/pharmacodynamic (PK/PD) parameters, such as the ratio of peak to minimum inhibitory concentration (peak/MIC ratio), ratio of 24-hour area under the curve to MIC (24-h AUC/MIC ratio), and the time above MIC, are good indicators of the drug dose-organism interaction . Time above the MIC is the important determinant of the activity of beta-lactams, macrolides, clindamycin, and linezolid . Free drug serum levels of these drugs should be above the MIC for at least 40%-50% of the dosing interval to produce adequate clinical and microbiological efficacy . Peak/MIC and 24-h AUC/MIC ratios are major determinants of the activity of aminoglycosides and fluoroquinolones . In general, peak/MIC ratios should exceed 8 and 24-h AUC/MIC values should be >100 to successfully treat gram-negative bacillary infections and to prevent the emergence of resistant organisms during therapy . The successful treatment of pneumococcal infections with fluoroquinolones and azithromycin appear to require 24-h AUC/MIC ratios of only 25-35 . Mutation prevention concentrations are being reported for various fluoroquinolones with different pathogens, but their clinical significance has not yet been established . More information is needed on the role of PK/PD parameters and their magnitude for preventing mutations and the emergence of resistant organisms for most classes of antibiotics.

Cancer Res, 2001 Aug 15, 61(16), 5974 - 8
Analysis of gene expression identifies candidate markers and pharmacological targets in prostate cancer; Welsh JB et al.; Detection, treatment, and prediction of outcome for men with prostate cancer increasingly depend on a molecular understanding of tumor development and behavior . We characterized primary prostate cancer by monitoring expression levels of more than 8900 genes in normal and malignant tissues . Patterns of gene expression across tissues revealed a precise distinction between normal and tumor samples, and revealed a striking group of about 400 genes that were overexpressed in tumor tissues . We ranked these genes according to their differential expression in normal and cancer tissues by selecting for highly and specifically overexpressed genes in the majority of cancers with correspondingly low or absent expression in normal tissues . Several such genes were identified that act within a variety of biochemical pathways and encode secreted molecules with diagnostic potential, such as the secreted macrophage inhibitory cytokine, MIC-1 . Other genes, such as fatty acid synthase, encode enzymes known as drug targets in other contexts, which suggests new therapeutic approaches.

Bioorg Med Chem, 2001 Aug, 9(8), 2203 - 11
Nickel(II) 2,6-diacetylpyridine bis(isonicotinoylhydrazonate) and bis(benzoylhydrazonate) complexes: structure and antimycobacterial evaluation . Part XI; Bottari B et al.; The reaction of 2,6-diacetylpyridine (dap) and isonicotinoyl- or benzoylhydrazide leads to bishydrazones H(2)dapin (1a) and H(2)dapb (1b), respectively . The condensation can either take place as a bimolecular kinetic process between the two reactants or as a monomolecular metal-templated synthesis in the presence of nickel(II) ions . In the latter case the reaction products are charged 2,6-diacetylpyridine bis(hydrazone) nickel(II) complexes, which can be easily deprotonated to neutral hydrazonates . Diffractometric analysis of one of these {Ni(dapb)}(2) (8b) has shown a binuclear structure with two octahedral nickel(II) ions bridged by two helicoidal dap (bishydrazonates) in a spheroidal structure of C(2V) symmetry . The synthesized complexes 8 are promising as antimycobacterial agents against M . tuberculosis H37Rv . In particular, 8b displays significant activity (MIC=0.025 microg/mL) 10-fold higher than rifampin and equal to isoniazid, while its ligand is ineffective . Compound 8b is also capable of reducing HIV-induced cytopathogenic effect in human T(4 )lymphocytes.

Antimicrob Agents Chemother, 2001 Sep, 45(9), 2609 - 15
DNA sequence analysis of rdxA and frxA from 12 pairs of metronidazole-sensitive and -resistant clinical Helicobacter pylori isolates; Kwon DH et al.; We previously reported that inactivation of rdxA and/or frxA converted Helicobacter pylori from metronidazole sensitive to metronidazole resistant . To examine the individual roles of rdxA and frxA in the development of metronidazole resistance in H . pylori, we examined the status of rdxA and frxA from 12 pairs of metronidazole-sensitive and -resistant H . pylori isolates obtained following unsuccessful therapy containing metronidazole . Arbitrary primed fingerprinting analyses revealed that the genotypes of 11 sensitive and resistant pairs of strains were essentially identical . Amino acid sequence identities of RdxA and FrxA from the 14 metronidazole-sensitive isolates ranged from 92 to 98% and 95 to 98%, respectively, compared to that of H . pylori J99 (MIC, 1 microg/ml) . All strains with high-level metronidazole resistance (MICs, 128 microg/ml) contained premature truncation of both RdxA and FrxA caused by nonsense and/or frameshift mutations . Strains with intermediate resistance to metronidazole (MICs, 32 to 64 microg/ml) contained a single premature truncation and/or altered RdxA and FrxA caused by nonsense, frameshift, and unique missense mutations . The low-level metronidazole-resistant strains (MICs, 8 microg/ml) contained unique missense mutations in FrxA but no specific changes in RdxA . The results demonstrate that alterations in both the rdxA and frxA genes are required for moderate and high-level metronidazole resistance and that metronidazole resistance that develops during anti-H . pylori therapy containing metronidazole is most likely to involve a single sensitive strain infection rather than a coinfection with a metronidazole-resistant strain.

Antimicrob Agents Chemother, 2001 Sep, 45(9), 2486 - 94
In vitro activities of fluoroquinolones against the spirochete Borrelia burgdorferi; Kraiczy P et al.; Little is known to date about the in vitro activity of fluoroquinolones against Borrelia species . Our study aimed at determining the in vitro activities of 15 quinolones against nine isolates of the Borrelia burgdorferi sensu lato complex in addition to one Borrelia valaisiana and one Borrelia bissettii tick isolate . For the determination of MICs, a standardized colorimetric microdilution method was applied . Determination of minimal borreliacidal concentrations providing 100% killing of the final inoculum (MBCs) after 72 h and time-kill experiments were performed by conventional culture in Barbour-Stoenner-Kelly medium in combination with dark-field microscopy . The rank order of potency on a microgram-per-milliliter basis for the substances with in vitro activity against B . burgdorferi was gemifloxacin (MIC at which 90% of the isolates tested are inhibited {MIC(90)}, 0.12 microg/ml) > sitafloxacin (MIC(90), 0.5 microg/ml), grepafloxacin (MIC(90), 0.5 microg/ml) > gatifloxacin (MIC(90), 1 microg/ml), sparfloxacin (MIC(90), 1 microg/ml), trovafloxacin (MIC(90), 1 microg/ml) > moxifloxacin (MIC(90), 2 microg/ml), ciprofloxacin (MIC(90), 2 microg/ml) > levofloxacin (MIC(90), 4 microg/ml) > ofloxacin (MIC(90), 8 microg/ml), norfloxacin (MIC(90), 8 microg/ml) > fleroxacin (MIC(90), >16 microg/ml), and pefloxacin (MIC(90), 32 microg/ml) > nalidixic acid (MIC(90), 256 microg/ml) . After 72 h of exposure, gemifloxacin was borreliacidal (100% killing) against the isolates investigated at a median MBC of 4 microg/ml . In the other compounds tested, median MBCs were higher (> or =8 microg/ml) . Results of electron microscopy and time-kill studies clearly support an in vitro activity of some fluoroquinolones against borreliae . Our study demonstrates for the first time the enhanced in vitro effectiveness of some of the recently introduced 4-quinolones against B . burgdorferi.

Heart Surg Forum, 2001, 4(1), 89 - 90
Closed chest coronary artery bypass on the beating heart; Kappert U et al.; Minimally invasive surgical procedures have become a part of routine cardiac surgery . The surgical techniques have been developed for the treatment of coronary artery disease in order to minimize surgical trauma . With the introduction of a 3-D-based totally endoscopically functioning system into minimally invasive cardiac (MIC) surgery, further reduction of skin incisions became possible and enhanced MIC techniques could be improved . Due to the 6 degrees freedom of motion allowed by wrist-enhanced instruments and a newly developed endoscopic stabilizer, totally endoscopic coronary artery bypass procedures on a beating heart became feasible . We present here our initial series of totally endoscopic "off-pump" coronary artery bypass grafting in patients suffering from coronary artery single vessel disease . In all patients, the procedure was successfully performed via four 1 cm chest incisions as closed-chest procedures.

Folia Microbiol (Praha), 2000, 45(6), 485 - 90
Effects of subinhibitory concentrations of nitroxoline on the surface properties of Escherichia coli; Latrache H et al.; Nitroxoline (5-nitro-8-quinolinol; NIQ) at subinhibitory concentrations (sub-MIC) decreased the adherence of uropathogenic Escherichia coli to catheter surface and significantly enhanced cell surface hydrophobicity . The surface hydrophobicity increased in the presence of sub-MIC of NIQ and also in an excess of Mg2+ . The effect of NIQ on the cell surface was not related to the bacteriostatic effect of this agent . The increase in nitrogen and decrease in phosphate content in the cell surface was found in the presence of NIQ . NIQ did not inhibit the expression of fimbriae.

Folia Microbiol (Praha), 2001, 46(2), 151 - 5
Occurrence of variants with temperature-dependent susceptibility (TDS) to antibiotics among Stenotrophomonas maltophilia clinical strains; Hejnar P et al.; Susceptibility to 20 antibiotics was tested in 104 Stenotrophomonas maltophilia strains at 37 and 30 degrees C by means of a dilution micromethod to verify the phenomenon of temperature-dependent susceptibility (TDS) . Trimethoprim-sulfamethoxazole, pefloxacin and ofloxacin were the most active preparations at 37 degrees C (93, 90, and 86% of susceptible strains, respectively), whilst trimethoprim-sulfamethoxazole, cefoperazone-sulbactam and pefloxacin performed best at 30 degrees C (94, 94, and 76% of susceptible strains, respectively) . Variants 37TDS (minimum inhibitory concentration, MIC, of tested antibiotics at least 4-times lower at 37 than at 30 degrees C) occurred in 60% . Variants 30TDS (at least 4-times lower value of MIC at 30 than at 37 degrees C) were found in 7.7% . Both variants in susceptibility to tested antibiotics appeared in 23.1%, whilst neither of them was observed in 9.6% . The 37TDS phenomenon was recorded most of all with gentamicin (51% of strains), amikacin (47), colistin (44) and tetracycline (34) . The 30TDS phenomenon was found particularly with cefoperazone-sulbactam (16.0% of strains) and colistin (10.0%) . The above phenomena may be due to changes in membrane permeability, temperature-dependent ribosomal changes, and insufficient adaptation to higher temperatures of some strains of the originally environmental species S . maltophilia.

Rev Esp Quimioter, 2000 Dec, 13(4), 408 - 11
{In vitro activity of ascorbic acid}; Lopez-Brea M et al.; The aim of this study was to determine the in vitro activity of ascorbic acid (vitamin C) against 43 Helicobacter pylori clinical isolates obtained from gastric biopsy taken by endoscopy . The MIC was determined by an agar dilution method . The ascorbic acid was included as two-fold dilutions in the agar to obtain a final concentration from 4000 mg/l to 62 mg/l . Brain-heart infusion agar supplemented with yeast extract and 10% fetal calf serum, 10(6) CFU/spot inoculum and microaerobic incubation for 2 to 5 days were used . MIC(50) 500 mg/l, MIC(90) 1000 mg/l and the range from 62 to 2000 mg/l . No differences in susceptible or resistant isolates were observed . Ascorbic acid showed good in vitro activity against the H . pylori clinical isolates tested in this study, which could explain some of the beneficial properties of vitamin C.

Fresenius J Anal Chem, 2001 Jul, 370(5), 647 - 53
Progress in multi-ion counting spark-source mass spectrometry (MIC-SSMS) for the analysis of geological samples; Jochum KP et al.; Spark source mass spectrometry (SSMS) has experienced important and significant improvements in nearly all analytical features by the use of a multiple ion counting (MIC) system . Two procedures have recently been developed to further increase the analytical capabilities of MIC-SSMS in geochemistry . These are a mathematical correction of interferences, which is often necessary for the ultra trace element analysis of Nb, Ta, Zr, Hf and Y, and the development of an autospark system to hold the total ion beam constant . New analytical data for geological samples, especially international reference materials, are presented using the improved MIC-SSMS technique . The data set consists of high precision and low abundance data for Zr, Nb and Y in depleted reference materials . The MIC-SSMS results are compared with those of conventional SSMS using photoplates for ion detection . The precision of the MIC-SSMS isotope ratio measurements (about 1%) is more than a factor of 3 better than that of conventional SSMS, as demonstrated by analyses of Hawaiian samples . Total uncertainties of MIC-SSMS concentration data including all sources of error are generally between 2 and 5% for concentrations higher than about 0.3 microg/g and about 10% for trace element abundances in the ng/g range.

Oncogene, 2001 Jul 5, 20(30), 4007 - 18
Change in gene expression subsequent to induction of Pnn/DRS/memA: increase in p21(cip1/waf1); Shi Y et al.; Pnn (PNN) is a nuclear and cell adhesion-related protein . Previous work has suggested that Pnn/DRS/memA is a potential tumor suppressor involved in the regulation of cell adhesion and cell migration . Using the ecdysone-inducible mammalian expression system, a stable inducible GFP-tagged human Pnn gene (PNNGFP) expressing 293 cell line was created (EcR293-PNNGFP) . Cells induced to express PNNGFP not only exhibited increased cell-cell adhesion but also exhibited changes in cell growth and cell cycle progression . cDNA array analyses, together with real time PCR, revealed that the effects of exogenously expressed Pnn on cellular behavior may be linked to the regulation of the expression of specific subset genes . This subset includes cell cycle-related genes such as p21(cip1/waf1), CDK4, CPR2; cell migration and invasion regulatory genes such as RhoA, CDK5, TIMP-1, MMP-7, and EMMPRIN; and MIC-1 . Concordant with previous observations of Pnn-induced phenotype changes, genes coding for epithelial associated processes and cell division controls were elevated, while those coding for increased cell motility and cellular reorganizations were downregulated . We utilized p21 promoter-luciferase reporter constructs and demonstrated that a marked stimulation of p21 promoter activity in 293 cells correlated with increased Pnn expression . Taken together, these data indicate that Pnn may participate in the regulation of gene expression, thereby, positively promoting cell-cell adhesion, and negatively affecting cell migration and cell proliferation.

Int J Surg Pathol, 2000 Jan, 8(1), 59 - 65
Epithelial Phenotype in Ewing's Sarcoma/Primitive Neuroectodermal Tumor; Vakar-Lopez F et al.; Neural differentiation is an integral component of Ewing's sarcoma/primitive neuroectodermal tumor (PNET), which exhibits a continuous spectrum from minimal to prominent neural phenotype . Differentiation of Ewing's sarcomas/PNETs along other lineages or the expression of an epithelial phenotype is less common and-if present-may cause diagnostic difficulties . In this study we evaluated the frequency of epithelial differentiation in formalin-fixed and paraffin-embedded tissues of 33 (22 primary and 11 metastatic) Ewing's sarcomas/PNETs by using an immunohistochemical assay with several antikeratin antibodies . Focal positivity for low- or high-molecular-weight keratins was documented in 18% of the cases, and diffuse coexpression of low- and high-molecular-weight keratins was observed in two cases . Expression of the MIC-2 gene product was documented in 94% of the tumors . The primitive neural phenotype as revealed by expression of either neuron-specific enolase or synaptophysin was observed in 30% of the cases, but coexpression of both neural markers was present in only 15% of the tumors . This study documents that, in addition to primitive neural differentiation, Ewing's sarcomas/PNETs frequently exhibit focal positivity for keratins, with rare strong diffuse coexpression of both low- and high-molecular-weight keratins . The findings indicate that the expression of an epithelial phenotype, at least in a focal fashion, is a relatively frequent finding in otherwise typical Ewing's sarcomas/PNETs . Int J Surg Pathol 8(1):59-65, 2000

Int J Syst Evol Microbiol, 2001 Jul, 51(Pt 4), 1587 - 92
Evolution of nuclear dualism in ciliates: a reanalysis in light of recent molecular data; Katz LA; Ciliates are defined by the presence of dimorphic nuclei . Following conjugation, both the functional macronucleus (MAC) and the transcriptionally inactive germline micronucleus (MIC) develop from a zygotic nucleus . As the MAC develops, germline chromosomes are processed by excision of internal sequences, fragmentation and amplification of the remaining chromosomes . The extent of processing varies among lineages and, in all but one class of ciliates, the resulting MACs divide by an unusual process termed 'amitosis' . Research on these chromosomal rearrangements, largely from studies of only a handful of taxa from two of the nine classes of ciliates, has failed to find evidence of homologous processing among ciliate lineages . This observation, coupled with the structural diversity of MAC genomes among ciliates, led to the hypothesis of multiple origins of at least two MAC properties: (1) the ability to divide and (2) the mechanisms underlying chromosomal processing . Applying this logic to a more inclusive analysis of ciliate lineages, where an even greater diversity of MAC structure is observed, increases the potential number of origins of these MAC characteristics . Here, it is proposed that a single origin of a relatively plastic mechanism underlying MAC development better explains the observed diversity in MAC structure and processing among ciliates . Such a mechanism is suggested by the demonstration of epigenetic effects during MAC development in Paramecium and Tetrahymena.

Southeast Asian J Trop Med Public Health, 2001 Mar, 32(1), 216 - 8
Stability of ceftazidime in normal saline solution after exposure to light; Jaruratanasirikul S et al.; Bactericidal activity of ceftazidime is determined by the time that concentrations in tissue and serum are above the MIC for the pathogens during the dosing interval . Thus, the most effective mode of administration of ceftazidime is continuous infusion . However, this agent is light sensitive which may result in instability when administered by this method without protection from light . Until now we have had no data to demonstrate the stability of this drug during continuous infusion . Therefore, the objective of this study was to provide such data . One gram of ceftazidime was mixed with 1,000 ml normal saline and exposed to two 36 watt fluorescence lights for 24 hours . The distance between ceftazidime solution and light source was 1 meter . Twenty samples (1 g-ceftazidime in normal saline) solution were evaluated . The mean ceftazidime concentrations in normal saline solution were decreased by only 1.69%, 4.44% and 7.19% after 6, 12 and 24 hours after exposure to light, respectively . Therefore, we conclude that the reduction of drug concentration was not considered to be significantly high, and this agent can be administered by continuous infusion.

J Colloid Interface Sci, 2001 Aug 15, 240(2), 552 - 558
Synthesis and Surface Properties of N-Alkyl-N-methylgluconamides and N-Alkyl-N-methyllactobionamides; Burczyk B et al.; Three series of nonionic N-alkylaldonamides, N-alkyl-N-methylgluconamides (Cn-MGA, Cn: n-C(10)H(21), n-C(12)H(25), n-C(14)H(29), n-C(16)H(33), and n-C(18)H(37)), N-alkyl-N-methyllactobionamides (Cn-MLA, alkyl as above-mentioned), and N-oleyl-N-methylglucon/lactobionamide, were synthesized in the reaction of an appropriate N-alkyl-N-methylamine with delta-D-glucolactone and lactobionic acid, respectively . Krafft temperatures of aqueous solutions and surface properties of these surfactants at 20 degrees C, i.e., surface excess concentration, Gamma(cmc), surface area demand per molecule, A(min), efficiency in surface tension reduction, pC(20), effectiveness in surface tension reduction, Pi(cmc), critical micelle concentration, CMC, and CMC/C(20) parameter as well as standard free energies of adsorption, DeltaG degrees (ads), and of micellization, DeltaG degrees (mic), were determined . It was shown that introduction of the methyl group to the amide nitrogen increased the solubility of the surfactants, which was confirmed by their Krafft temperatures . Lactobionamides are more water soluble than gluconamides . On the other hand, the Cn-MGA surfactants are more surface active than the respective Cn-MLA ones . This observation is based on the determined adsorption and micellization parameters . The presence of one double bond in a hydrocarbon chain as in oleyl-amides increases their hydrophilic character compared with that of saturated C18 derivatives . No distinct differences were observed between the A(min) values obtained for both series studied, although they differ markedly in the size of the hydrophilic groups .

J Antimicrob Chemother, 2001 Aug, 48(2), 295 - 7
A 9 year study of clarithromycin and metronidazole resistance in Helicobacter pylori from Spanish children; Lopez-Brea M et al.; The evolution of clarithromycin, metronidazole and amoxycillin resistance in 246 Spanish Helicobacter pylori clinical isolates, obtained from paediatric patients during a 9 year period, was determined by an agar dilution technique . Clarithromycin resistance (MIC 1 mg/L) was 2.27% (IC95 0.05-12.02) in 1991-1993, 20.98% (IC95 12.72-31.46) in 1994-1996 and 28.33% (IC95 20.48-37.28) in 1997-1999 (P < 0.01) . Metronidazole resistance (MIC 8 mg/L) was 7.14% (IC95 1.49-19.48) in 1991-1993, 20.25% (IC95 12.04-30.79) in 1994-1996 and 43.90% (IC95 32.95-55.30) in 1997-1999 (P < 0.01) . Amoxycillin resistance was not found (all strains showed MICs < 2 mg/L).

J Antimicrob Chemother, 2001 Aug, 48(2), 259 - 67
Pharmacokinetic and pharmacodynamic profile of high dose extended interval piperacillin-tazobactam; Kim MK et al.; A multiple-dose, open-labelled, randomized, two period crossover human volunteer study was performed (i) to describe the pharmacokinetic profile and safety profile of piperacillin and tazobactam (P/T) administered 6.0/0.75 g and 8.0/1.0 g q12h and (ii) to characterize the pharmacodynamic profile of these regimens against a variety of common targeted pathogens . Blood samples were collected after the third dose and concentrations of P/T were determined by a validated high-performance liquid chromatography assay . Pharmacokinetic profiles of P/T were determined by non-compartment analysis . Percentage time above the MIC (%T > MIC) of piperacillin was calculated for a range of MICs . In this study, no adverse events were attributed after multiple administrations of either 6.0/0.75 g or 8.0/1.0 g dose regimens . The peak concentration, half-life and area under the curve (AUC0-(0-tau)) of piperacillin were significantly different by a paired t-test (P < 0.05) between the two study regimens . The trough concentration, half-life and area under the curve (AUC0-(0-tau)) of tazobactam were substantially different from parameters reported previously for conventional regimens . The 8.0/1.0 g regimen provided 50% T > MIC for MICs < or =32 mg/L, while a similar value for the 6.0/0.75 g regimen was < or = 16 mg/L . High-dose P/T regimens with extended interval were well tolerated and provide adequate dynamic exposure for a variety of susceptible pathogens.

Nippon Ishinkin Gakkai Zasshi, 2001, 42(3), 119 - 22
{A case of chromomycosis arising in a Japanese-Brazilian patient}; Nagao K et al.; A 41-year-old Japanese-Brazilian man, living in Japan since 1991, visited our hospital in August 1998 complaining of a scaly annular erythema which had been present on his right forearm since 1996 . Granulomatous inflammation was revealed in the dermis upon skin biopsy . Sclerotic cells were present within the granulomatous lesions . Fonsecaea pedrosoi was isolated from tissue cultures . The restriction fragment length polymorphism pattern of mitochondrial DNA of the causative fungus was compatible with F . pedrosoi type 1, the commonest type in Japan . Since type 4 is usually found in South America, the patient was assumed to have become infected in Japan . An oral dose of terbinafine at 125 mg/day was ineffective . 125 mg/day single dose of terbinafine reaches an average plasma concentration of only 0.69 g/ml, where as 250 mg/day reaches 0.96 micro g/ml . Because the minimum inhibitory concentration (MIC) of terbinafine against the isolated fungus was found to be 0.76 micro g/ml, the terbinafine dose was increased from 125 mg/day to 250 mg/day, which slowly led to remission . Chromomycosis is caused by several species of dematiaceous fungi, and terbinafine efficacy may vary depending on the causative fungus . MIC values may differ even within the same species . It is important to isolate the pathogenic fungus in each case and measure the MIC value to determine the optimal dose of terbinafine.

Zhonghua Jie He He Hu Xi Za Zhi, 1998 Oct, 21(10), 617 - 9
{Comparison of three antituberculous drugs in serum and cold abscesses of patients with spinal tuberculosis}; Wu Q et al.; OBJECTIVE: Pharmacokinetics of INH, RFP and OFLX in serum and cold abscesses of patients with spinal tuberculosis was analyzed to provide reference to choosing clinical therapeutic regimen . METHOD: The aspiration specimens of abscesses and venous blood were collected from 8 patients with spinal tuberculosis at 0.5, 0.75, 1.0, 1.5, 2.0, 4.0, 6.0, 9.0, 12.0, 16.0 and 24.0 h after administration of antituberculous drugs . The specimens were assessed by high performance liquid chromatography (HPLC) . The data were processed with software 3P87 . RESULT: The Cmax of INH, RFP and OFLX in serum were 10.87 +/- 7.09 micrograms/ml, 9.98 +/- 3.53 micrograms/ml, and 5.29 +/- 0.72 micrograms/ml, while the Cmax of INH, RFP and OFLX in cold abscesses were 2.84 +/- 1.63 micrograms/ml, 0.57 +/- 0.26 microgram/ml and 3.19 +/- 1.29 micrograms/ml respectively . CONCLUSION: After administration, the Cmax of INH and OFLX in the cold abscesses of patients with spinal tuberculosis, reached the level beyond their MIC and appeared and disappeared more slowly than that in the serum; RFP was not easy to permeate into the cold abscesses, the Cmax of RFP in the cold abscesses just reached its MIC.

Arch Pharm (Weinheim), 2001 Jun, 334(6), 189 - 93
Potential tuberculostatic agents: micelle-forming copolymer poly(ethylene glycol)-poly(aspartic acid) prodrug with isoniazid; Silva M et al.; With the objective of obtaining slow-acting isoniazid derivatives, of potential use as chemoprophylactics or chemotherapeutics in tuberculosis, the micelle-forming copolymer of poly(ethylene glycol)-poly(aspartic acid) prodrug with isoniazid was synthesized . The derivative obtained was found to be active in Mycobacterium tuberculosis culture, with a minimal inhibitory concentration (MIC) 5.6 times lower than that of the tuberculostatic drug.

Mycopathologia, 2001, 150(3), 101 - 15
In vitro antifungal activities of voriconazole and reference agents as determined by NCCLS methods: review of the literature; Espinel-Ingroff A et al.; Voriconazole (VfendTM) is a new triazole that currently is undergoing phase III clinical trials . This review summarizes the published data obtained by NCCLS methods on the in vitro antifungal activity of voriconazole in comparison to itraconazole, amphotericin B, fluconazole, ketoconazole and flucytosine . Voriconazole had fungistatic activity against most yeasts and yeastlike species (minimum inhibitory concentrations {MICs} < 2 microg/ml) that was similar or superior to those of fluconazole, amphotericin B, and itraconazole . Against Candida glabrata and C . krusei, voriconazole MIC ranges were 0.03 to 8 and 0.01 to > 4 microg/ml, respectively . For four of the six Aspergillus spp . evaluated, voriconazole MICs (< 0.03 to 2 microg/ml) were lower than amphotericin B (0.25 to 4 microg/ml) and similar to itraconazole MICs . Voriconazole fungistatic activity against Fusarium spp . has been variable . Against E oxysporum and F . solani, most studies showed MICs ranging from 0.25 to 8 microg/ml . Voriconazole had excellent fungistatic activity against five of the six species of dimorphic fungi evaluated (MIC90s < 1.0 microg/ml) . The exception was Sporothrix schenckii (MIC90s and geometric mean MICs > or = 8 microg/ml) . Only amphotericin B had good fungistatic activity against the Zygomycetes species (voriconazole MICs ranged from 2 to > 32 microg/ml) . Voriconazole showed excellent in vitro activity (MICs < 0.03 to 1.0 microg/ml) against most of the 50 species of dematiaceous fungi tested, but the activity of all the agents was poor against most isolates of Scedosporium prolificans and Phaeoacremonium parasiticum (Phialophora parasitica) . Voriconazole had fungicidal activity against most Aspergillus spp., B . dermatitidis, and some dematiaceous fungi . In vitro/in vivo correlations should aid in the interpretation of these results.

Ultrastruct Pathol, 2001 May-Jun, 25(3), 219 - 25
Immunohistochemical and ultrastructural investigation of neural differentiation in Ewing sarcoma/PNET of bone and soft tissues; Franchi A et al.; The authors evaluated the role of immunohistochemistry and electron microscopy in defining neural differentiation in 28 cases of Ewing sarcoma/PNET . The panel of primary antibodies used included vimentin, MIC-2, NSE, S-100 protein, leu7, neurofilaments, GFAP, and chromogranin A . Cases were considered undifferentiated when neural markers were absent, poorly differentiated if one neural marker was present, and well differentiated if two or more markers were observed . Cases were also evaluated for the presence of cytoplasmic processes, microtubules, and neurosecretory granules as ultrastructural features of neural differentiation: the tumor was classified as well differentiated if two of these features were present; and poorly differentiated if one was evident; all other cases were considered undifferentiated . According to immunohistochemistry, 10 cases (35.7%) were undifferentiated, 12 cases (42.9%) were poorly differentiated, and 6 (21.4%) were well differentiated . According to the ultrastructural analysis, 10 tumors were undifferentiated (35.7%), 14 poorly differentiated (50%), and 4 well differentiated (14.3%) . The overall concordance between the two techniques was low (35.7%), and both modalities were concordant in classifying only 1 well-differentiated, 5 poorly differentiated, and 4 undifferentiated tumors . In conclusion, the authors suggest that investigations devoted to test the prognostic significance of neural differentiation in these neoplasms should employ both immunohistochemistry and electron microscopy, separately and in combination, to assess what is the most effective choice for predicting the clinical course.

Int J Antimicrob Agents, 2001 Jul, 18(1), 73 - 6
Activity of the ketolide telithromycin (HMR-3647) against erythromycin-susceptible and -resistant pneumococci isolated in the UK; Johnson AP et al.; The novel ketolide telithromycin (formerly HMR-3647) was tested against a collection of pneumococci of varying sensitivity to erythromycin and clindamycin, isolated in geographically diverse UK hospitals . Telithromycin was highly active against erythromycin-susceptible pneumococci, the MIC(90) being 0.015 mg/l . Erythromycin-resistant pneumococci that contained the ermB gene, either alone or together with the mefE gene, were cross-resistant to other macrolides and to clindamycin, while erythromycin-resistant pneumococci that contained only the mefE gene were cross-resistant to azithromycin, clarithromycin and roxithromycin but remained susceptible to josamycin and clindamycin . Telithromycin was active against erythromycin-resistant pneumococci irrespective of their mechanism of macrolide resistance, although the MIC(90) (0.25 mg/l) was higher than that seen with erythromycin-sensitive isolates . Telithromycin thus appears to be a potentially useful drug in settings where pneumococcal resistance to macrolides is prevalent.

J Infect Chemother, 2001 Jun, 7(2), 59 - 68
Clinical observation and treatment of leptospirosis; Kobayashi Y; The epidemiological and clinical observations of 240 patients with Weil's disease and 10 patients with canicola fever, and these observations in two epidemics of canicola fever, are presented . Early diagnosis is most important for the prognosis of patients with the severe form of leptospirosis . It depends on the clinical features, clinical laboratory findings, and the epidemiological situation . The most characteristic clinical signs for early diagnosis were febrile illness of sudden onset, severe general malaise, muscular pain, and conjunctival congestion . Proteinuria, leukocytosis with neutrophilia, and raised erythrocyte sedimentation rate were the most indicative clinical laboratory findings for early diagnosis . Although jaundice and hemorrhage are the most important signs of the severe form of leptospirosis, Weil's disease, these are rarely useful in early diagnosis . Of a variety of antibiotics used, penicillins and cephems had the lowest minimal inhibitory concentration against leptospires . However, it became apparent from basic studies in vitro and in vivo that streptomycin showed the best bactericidal action against leptospires and that it was the most effective anti-leptospiral antibiotic . Gentamicin, tobramycin, and isepamicin are also effective as alternatives to streptomycin . Although penicillins, cephems, tetracyclines, and macrolides are also effective for the treatment of leptospirosis, when these antibiotics with inadequate bactericidal activity are used for the treatment of the disease, long-term therapy with sufficiently large doses may be required from an early stage of the disease until the appearance of antibodies.

Bioorg Med Chem Lett, 2001 Aug 6, 11(15), 1965 - 9
Antimalarial halorosellinic acid from the marine fungus Halorosellinia oceanica; Chinworrungsee M et al.; Three known compounds, 2-hexylidene-3-methylsuccinic acid (1), cytochalasin Q (2), and 5-carboxymellein (3), together with two new derivatives, 2-hexylidene-3-methylsuccinic acid 4-methyl ester (4) and an ophiobolane sesterterpene named halorosellinic acid (5), were isolated from culture broth of the marine fungus Halorosellinia oceanica BCC 5149 . Compounds 1-3 exhibited moderate cytotoxicity against KB and BC-1 cell lines with IC(50) values of 1-13 microg/mL, while compounds 2, 3, 5, and 6 showed antimalarial activity with respective IC(50) values of 17, 4, 13, and 19 microg/mL . Halorosellinic acid (5) possessed only weak antimycobacterial activity with the minimum inhibitory concentration of 200 microg/mL.

Br J Clin Pharmacol, 2001 Jul, 52(1), 65 - 8
Diurnal variation in the biliary excretion of flomoxef in patients with percutaneous transhepatic biliary drainage; Hishikawa S et al.; AIMS: To examine diurnal variation in biliary excretion of flomoxef . METHODS: Flomoxef (1 g) was injected intravenously in eight patients with percutaneous transhepatic cholangiography with drainage at 09.00 h and 21.00 h by a cross-over design with a 36 h washout period . Drained biliary fluid was collected for 6 h after each dosing . These patients still had mild to moderate hepatic dysfunction . RESULTS: Bile flow and bile acid excretion for 6 h after dosing did not differ significantly between the 09.00 h and 21.00 h treatments . The maximum concentration of biliary flomoxef was significantly greater and its total excretion for 6 h tended to be greater after the 21.00 h dose {maximum concentration (microg ml(-1)): 34.2 +/- 29.9 (09.00 h dose) vs 43.5 +/- 28.3 (21.00 h dose) (95% confidence interval for difference: 2.6 approximately 15.9, P = 0.013); total excretion (mg 6 h(-1)): 1.4 +/- 1.3 (09.00 h dose) vs 1.6 +/- 1.2 (21.00 h dose) (95% confidence interval for difference: -26.8, 313.7, P = 0.087)} . The period that biliary flomoxef remained above the minimal inhibitory concentration did not differ significantly between the two treatment times . CONCLUSIONS: These results suggest that biliary excretion of flomoxef shows diurnal variation . However, as the difference was relatively small, flomoxef could be given at any time of day without any dosage adjustments.

Chem Res Toxicol, 2001 Jul, 14(7), 833 - 40
N-methylcarbamoylated valine of hemoglobin in humans after exposure to N,N-dimethylformamide: evidence for the formation of methyl isocyanate?
Kafferlein HU, Angerer J.
N,N-Dimethylformamide (DMF) is reported to cause testicular germ-cell tumors in exposed workers . The reports, however, are not in line with results obtained in animal and in vitro experiments, where DMF was shown not to be mutagenic and also not to be carcinogenic . Considerable interest raised on the formation of a reactive intermediate, presumably methyl isocyanate (MIC), during metabolism of DMF in humans over the last years . We report the formation of N-methylcarbamoylated valine of hemoglobin (Hb) in blood samples from workers exposed to DMF in the polyacrylic fiber industry . N-Methylcarbamoylated Hb was formed by the reaction of MIC with Hb . For this purpose, Hb adducts were monitored by means of a modified Edman degradation involving the release of the N-terminal valine adduct in form of 3-methyl-5-isopropylhydantoin (MIH) . For internal standardization of the method, 3-ethyl-5-isopropylhydantoin (EIH) was used . Separation and analysis of MIH and EIH were carried out by gas chromatography and mass spectrometry with electron impact ionization (GC/EI-MS) . Hb adducts in form of MIH were quantified in blood samples from exposed persons in concentrations between 26.1 and 412.0 nmol of MIH/g of globin . The observed adducts were proven to be identical to those derived from the in situ reaction between Hb and MIC . Taken together with the fact that only N-methylcarbamoylated Hb can undergo ring closure to the corresponding hydantoin, the reaction is indirect evidence for the occurrence of MIC in vivo . The formation of MIC directly in the cell and its possible distribution through the human body may lead to critical effects after exposure to DMF . Adducts were determined not to be totally specific for exposure to DMF since an identical adduct was also found in blood samples from the general population . However, concentrations were lower by a factor of about 100 . The sources for background adducts are currently unknown.

Eur J Clin Microbiol Infect Dis, 2001 May, 20(5), 329 - 33
Characterization of isoniazid-resistant strains of Mycobacterium tuberculosis on the basis of phenotypic properties and mutations in katG; Abate G et al.; Forty isoniazid-resistant Mycobacterium tuberculosis isolates were characterized on the basis of phenotypic properties (i.e., catalase activity, MIC of isoniazid, and growth pattern in the presence of 7 different concentrations of isoniazid) and alterations in the katG gene (codons 315 and 463) . Three different growth patterns could be distinguished: concentration-dependent inhibition of growth was observed in 29 strains, similar growth at all concentrations was seen in 7 strains, and enhanced growth at low concentrations of isoniazid was evident in 4 strains . The MIC of isoniazid was < or = microg/ml for 29 of 40 strains . Mutation at codon 315 of the katG was detected in 28 of 40 strains . However, only one of the seven strains for which the MIC of isoniazid was > or = 16 microg/ml had mutation at this codon . Five of these seven strains for which the MIC was > or = 16 microg/ml had no catalase activity . The results indicate that the MIC of isoniazid for a majority of strains is below the level achievable in serum . Therefore, isoniazid may be beneficial for the treatment of some cases of multidrug-resistant tuberculosis . Determination of catalase activity aids in the detection of isolates for which MICs are high and could, in conjunction with molecular methods, provide rapid detection of most isoniazid-resistant strains.

Eur J Clin Microbiol Infect Dis, 2001 May, 20(5), 299 - 308
Clinical characteristics and outcome of patients with invasive pneumococcal disease, Puy-de-Dôme, France, 1994-1998; Laurichesse H et al.; A surveillance program for invasive pneumococcal disease was undertaken in Puyde-Djme, an administrative district of the region Auvergne in France, from 1 January 1994 to 31 December 1998 . A total of 214 cases were identified . The annual incidence of invasive pneumococcal disease increased (P=0.04) from 5.5 in 1994 to 9.3 cases per 10(5) person-years in 1998 . The highest incidences were for children <2 years of age (59.2 cases per 10(5) person-years) and for adults > or = 65 years (18 cases per 10(5) person-years) . Clinical diagnoses, available in 200 patients, included acute pneumonia (62%), meningitis (10%), sepsis without focus (20%), and others (8%) . The most frequent chronic medical conditions of the patients included smoking, alcoholism, cardiovascular and pulmonary diseases, and malignancies . Thirty-one percent of the isolates were nonsusceptible to penicillin . Penicillin resistance (MIC > or = 0.1 mg/l) was more frequent (P=0.02) in cancer patients . The overall case-fatality rate was 21.5% . Risk factors for death were age, sex, and underlying diseases of the patients, along with the severity of illness . These population-based findings should convince clinicians to offer pneumococcal vaccine to patients at high risk for invasive pneumococcal disease, thereby increasing vaccination coverage levels in France.

Med Pediatr Oncol, 2001 Feb, 36(2), 290 - 4
Ewing sarcoma and sinonasal neuroectodermal tumors as second malignant tumors after retinoblastoma and other neoplasms; Cope JU et al.; BACKGROUND: Excesses of various childhood cancers have been reported after retinoblastoma, including a trickle of Ewing sarcoma (ES) and perhaps histologically similar olfactory neuroblastoma, both of which are neural tumors . To update and advance this information, case reports were sought by an extensive review of the literature . PROCEDURE: The search was made through the use of PubMed, and the Web of Science (Citation Index Expanded), keying on primary references . Three sinonasal cancers diagnosed as ES were immunohistochemically stained for MIC-2 protein (positive in ES) . RESULTS: Retinoblastoma occurred before ES in ten cases (seven bilateral) . In four others, retinoblastoma (three bilateral) developed before sinonasal neural tumors (poorly differentiated) . ES also occurred after 14 cancers other than retinoblastoma (five lymphomas, four leukemias, and one each of five miscellaneous cancers) . The predominance of retinoblastoma prior to ES differs markedly from the low-frequency of retinoblastoma among childhood cancers in the general population . On the contrary, cancers other than retinoblastoma were proportionate to those in the general population . Previously, retinoblastoma followed by excesses of osteosarcoma and soft tissue sarcomas has been attributed to the action of the inherited RB-1 gene . The sinonasal tumors stained negative for MIC-2 protein . CONCLUSIONS: Heritable retinoblastoma may predispose to ES and perhaps to a subset of poorly differentiated neuroectodermal tumors in the sinonasal region that may be related to olfactory neuroblastoma.

Antimicrob Agents Chemother, 2001 Aug, 45(8), 2210 - 4
Telithromycin is active against Mycobacterium avium in mice despite lacking significant activity in standard in vitro and macrophage assays and is associated with low frequency of resistance during treatment; Bermudez LE et al.; The activity of telithromycin, a new ketolide, was evaluated in vitro and in vivo against Mycobacterium avium complex (MAC) strains . The MIC of telithromycin for several M . avium isolates obtained from the blood of AIDS patients ranged from 16 to >128 microg/ml (MIC at which 90% of isolates are inhibited, >128 microg/ml), and the compound did show activity in the macrophage system at concentrations greater than 8 or 16 microg/ml, but this was dependent on the MAC strain used . Telithromycin was then administered to mice infected with MAC strain 101 for 4 weeks at doses of 100, 200, or 400 mg/kg of body weight/day . Treatment with 100 and 200 mg/kg/day was bacteriostatic, but at 400 mg/kg/day telithromycin was bactericidal for MAC strains . The frequency of the emergence of resistance to telithromycin was low despite prolonged usage (12 weeks) . This study demonstrates that telithromycin is active in vivo against MAC and warrants further evaluation.

J Colloid Interface Sci, 2001 Aug 1, 240(1), 375 - 377
Enthalpies of Micellization of Double Chain and Gemini Cationic Surfactants; Bai G et al.; Microcalorimetric measurements have been made on double chain cationic surfactants with the formula {C(N)H(2N+1)C(M)H(2M+1)N(CH(3))(2)}Br, referred to as C(N)C(M)DAB, and a cationic gemini surfactant with the formula {C(12)H(25)(CH(3))(2)N(CH(2))(12)N(CH(3))(2)C(12)H(25)}Br(2), referred to as C(12)C(12)C(12)Br(2) . The CMCs and enthalpies of micellization for the compounds C(12)C(6)DAB, C(14)C(4)DAB, C(16)C(2)DAB, and C(12)C(12)C(12)Br(2) are obtained from experiments . The effect of the ratio (N/M) of the two alkyl chain lengths for the C(N)C(M)DABs on the micellization process has been investigated . The contribution of DeltaH(mic) to DeltaG(mic) is about 3.4% at N/M=2, about 11% at N/M=3.5 and about 39% at N/M=8, suggesting that when the total carbon atoms (N+M) remain constant, the micellization process will be driven by entropy or entropy and enthalpy jointly with increasing ratio (N/M) . In addition, the comparison between C(12)C(12)C(12)Br(2) and its corresponding monomer (C(12)C(6)DAB) is interesting . The micellization process for C(12)C(12)C(12)Br(2) is driven jointly by enthalpy and entropy, whereas the micellization process for C(12)C(6)DAB is driven mainly by entropy .

J Colloid Interface Sci, 2001 Aug 1, 240(1), 272 - 276
The Behavior of Sorbitan Surfactants at the Water-Oil Interface: Straight-Chained Hydrocarbons from Pentane to Dodecane as an Oil Phase; Peltonen L et al.; The interfacial tension of four sorbitan surfactants (Span 20, sorbitan monolaurate; Span 40, sorbitan monopalmitate; Span 60, sorbitan monostearate; and Span 80, sorbitan monooleate) was determined at the water-oil interface . Seven straight-chained hydrocarbons from pentane to dodecane were used as an oil phase . From the interfacial tension measurements the following values were calculated: critical micelle concentration (cmc), the interfacial tension at the cmc (gamma(cmc)), surface pressure at the cmc (pi(cmc)), area per molecule at the cmc (A(cmc)), standard free energy of micellization (DeltaG degrees (mic)), and standard free energy of adsorption (DeltaG degrees (ad)) . The shorter chained Span 20 and unsaturated Span 80 had higher cmc values and Span 80 had a larger molecular area than the other surfactants . With the same oil phase, differences between pi(cmc) values of the four sorbitan monoesters were small, but the gamma(cmc) was slightly lowered as the hydrophobicity of the surfactant was increased . DeltaG degrees (mic) was less negative for Span 20 and the DeltaG degrees (ad) value was slightly more negative for Span 80 . The effect of the oil phase was obvious . Increasing the hydrocarbon chain length of the oil phase increased gamma(cmc) and cmc values while pi(cmc) and A(cmc) were decreased . As the length of the hydrocarbon chain of the oil phase was increased, DeltaG degrees (mic) and DeltaG degrees (ad) became less negative, which means a less spontaneous reaction .

Pharmacotherapy, 2001 Jul, 21(7), 855 - 60
Cunninghamella bertholletiae infection in a bone marrow transplant patient: amphotericin lung penetration, MIC determinations, and review of the literature; Garey KW et al.; Infections caused by Cunninghamella bertholletiae, an opportunistic fungal organism, have an extremely high mortality rate . A fatal case of C . bertholletiae fungal pneumonia occurred in a man who had received an allogeneic bone marrow transplant . Aggressive debridement and high-dose liposomal amphotericin B failed to eradicate the infection . Right lung tissue samples obtained during lobectomy were assayed for amphotericin B concentrations by high-performance liquid chromatography, and minimum inhibitory concentration (MIC) determinations of amphotericin B against C . bertholletiae were determined by the macrobroth dilution method . The MIC for the isolate of C . bertholletiae was 4 microg/ml . Amphotericin B lung concentrations averaged 9.5 microg/ml (range 3.7-13.8 microg/ml), with a corresponding serum trough concentration of 0.9 microg/ml . To our knowledge, this is the first reported case of amphotericin B concentrations measured at the site of infection in a patient with a pulmonary Cunninghamella infection, together with a corresponding MIC of the organism . The patient's death, which occurred despite aggressive debridement and high amphotericin B lung concentrations, highlights the need for novel strategies to treat infections caused by invasive molds such as C . bertholletiae.

Clin Microbiol Infect, 2001 Jun, 7(6), 326 - 30
Interpretive criteria of ceftibuten disk diffusion susceptibility tests according to the DIN 58 940 method; Kleinkauf N et al.; OBJECTIVE: This study aimed to establish interpretive criteria for agar diffusion tests with ceftibuten disks according to DIN standards . METHODS: Minimal inhibitory concentrations (MICs) and inhibition zones produced by ceftibuten in the disk diffusion test were determined for 275 recent bacterial isolates, including 11 species with 25 strains each . Regression analysis was performed for two disk loads (10 microg and 30 microg) . RESULTS: Correlation of MICs and zone diameters was good, with correlation coefficients of r = - 0.97 for both tested disk loads . Evaluation of the calculated zone size criteria for all species showed no very major discrepancies or no major discrepancies . The 30-microg disks, however, produced unacceptably large inhibition zones for very susceptible strains, so that usage of 10-microg disks must be recommended when testing according to DIN standards . CONCLUSION: Based on the MIC breakpoints recommended by the DIN (> or =8 mg/L and < or = 1 mg/L), the following interpretive breakpoints for disk diffusion susceptibility tests with 10-microg ceftibuten disks were calculated using regression line analysis: < or =19 mm for resistance and > or = 27 mm for susceptiblity . Proposed inhibition zone diameters for the reference strain Escherichia coli ATCC 25922 are between 31 and 36 mm.

J Chemother, 2001 Apr, 13 Suppl 1, 18 - 22
Trachoma: a review; Tabbara KF; Trachoma is a leading cause of preventable blindness worldwide . The disease is caused by an intracellular epithelial gram-negative bacterium, Chlamydia trachomatis . The presence of children, overcrowding, and the lack of water in the household are factors that predispose to the transmission of the disease . The disease may remain asymptomatic but some patients many complain of redness, irritation, and ocular discharge . The principal initial clinical manifestation is a follicular conjunctivitis that may lead to conjunctival scarring, entropion, trichiasis, corneal thinning, and ulceration . Some patients develop corneal scars that lead to loss of vision . Despite the remarkable progress in our understanding of Chlamydial infections, the basic mechanisms involved in tissue damage and scarring remain to be elucidated . There are several effective therapeutic modalities for trachoma . Azithromycin oral single dose was found to be safe and effective in children with active trachoma . Conjunctival biopsy specimens obtained from adult patients receiving a single oral dose of azithromycin showed sustained high levels of azithromycin (above MIC of chlamydia) for up to 2 weeks after intake . These prolonged high levels of azithromycin in the conjunctival tissue following a single oral dose makes the drug suitable for the treatment of endemic trachoma.

Nippon Saikingaku Zasshi, 2001, 56(2), 455 - 63
{Activity of bicozamycin against Escherichia coli O157:H7 producing Vero toxin}; Matsumoto Y; Bicozamycin (BCM) which inhibits protein synthesis by inhibiting Rho-dependent transcription termination factor ATPase activity is used for treatment of animal infections . BCM had moderate activity of MIC 16-32 micrograms/ml against enterohemorrhagic Verotoxin (VT)-producing Escherichia coli (EHEC) O157:H7 and inhibited production of VT1 and VT2 . The activity of BCM against EHEC was slightly higher in anaerobic conditions, and was more evident in vivo . BCM decreased CFUs of EHEC in caecum more effectively than fosfomycin, cefixime and norfloxacin in a mouse infection model . Moreover, BCM did not increase the amount of either VT1 or VT2 in caecum in mice . In contrast, norfloxacin increased mortality of mice infected with EHEC by inducing VT production . The results suggest that BCM is useful for the treatment of EHEC infection and eradication of EHEC . Dairy live stock, especially young animals, have been implicated as a principal reservoir of EHEC . Eradication of EHEC from live stock will become an important problem in the near future . The narrow spectrum antibiotic BCM is expected to be a safe and effective antibiotic to eradicate EHEC from live stock.

Trends Immunol, 2001 Jul, 22(7), 378 - 85
MICA and MICB genes: can the enigma of their polymorphism be resolved?
Stephens HA.
The human MHC class I chain-related genes (MICA and MICB) are located within the HLA class I region of chromosome 6 . Their organization, expression and products differ considerably from classical HLA class I genes . MIC proteins are considered to be markers of "stress" in the epithelia, and act as ligands for cells expressing a common activatory natural killer-cell receptor (NKG2D) . Molecular models are now available for the MICA protein, both bound and complexed with NKG2D . MICA molecules appear to be highly flexible and polymorphic, although the functional relevance and implications of their polymorphism have yet to be fully discerned.

J Clin Microbiol, 2001 Jul, 39(7), 2458 - 62
Antifungal susceptibility testing of fluconazole by flow cytometry correlates with clinical outcome; Wenisch C et al.; Susceptibility testing of fungi by flow cytometry (also called fluorescence-activated cell sorting {FACS}) using vital staining with FUN-1 showed a good correlation with the standard M27-A procedure for assessing MICs . In this study we determined MICs for blood culture isolates from patients with candidemia by NCCLS M27-A and FACS methods and correlated the clinical outcome of these patients with in vitro antifungal resistance test results . A total of 24 patients with candidemia for whom one or more blood cultures were positive for a Candida sp . were included . Susceptibility testing was performed by NCCLS M27-A and FACS methods . The correlation of MICs (NCCLS M27-A and FACS) and clinical outcome was calculated . In 83% of the cases, the MICs of fluconazole determined by FACS were within 1 dilution of the MICs determined by the NCCLS M27-A method . For proposed susceptibility breakpoints, there was 100% agreement between the M27-A and FACS methods . In the FACS assay, a fluconazole MIC of <1 microg/ml was associated with cure (P < 0.001) whereas an MIC of > or =1 microg/ml was associated with death (P < 0.001) . The M27-A-derived fluconazole MICs did not correlate with outcome (P = 1 and P = 0.133).

Bioorg Med Chem Lett, 2001 Jul 9, 11(13), 1703 - 7
Some 3-thioxo/alkylthio-1,2,4-triazoles with a substituted thiourea moiety as possible antimycobacterials; Kucukguzel I et al.; A series of novel N-alkyl/aryl-N'-{4-(4-alkyl/aryl-2,4-dihydro-3H-1,2,4-triazole-3-thione-5-yl)phenyl}thioureas 1-19 and three S-alkylated representatives of the former, N-alkyl/aryl-N'-{4-(3-aralkylthio-4-alkyl/aryl-4H-1,2,4-triazole-5-yl)phenyl}thioureas 20-22, were synthesized and tested for antimycobacterial activity against Mycobacterium tuberculosis H37Rv as well as Mycobacterium fortuitum ATCC 6841 which is a rapid growing opportunistic pathogen . Compounds 4 and 9-11 were found to possess the same MIC value with that of Tobramycin against M . fortuitum ATCC 6841 whereas 1-3 and 21 had positive response against M . tuberculosis H37Rv at varying degrees . Compound 21 was identified as the most potent derivative of the 1-22 series by an MIC value of 6.25 microg/mL and selectivity index of 1.6.

Bioorg Med Chem Lett, 2001 Jul 9, 11(13), 1675 - 8
Antimycobacterial activity of 9-sulfonylated/sulfenylated-6-mercaptopurine derivatives; Scozzafava A et al.; A series of 9-sulfonylated/sulfenylated-6-mercaptopurines has been prepared by reaction of 6-mercaptopurine with sulfonyl/sulfenyl halides . These compounds constitute a new class of potent antimycobacterial agents, possessing MIC values against Mycobacterium tuberculosis H37Rv in the range of 0.39-3.39 microg/mL, as well as appreciable activity against Mycobacterium avium . Furthermore, a compound of this small series exhibited good activity (MIC under 1 microg/mL) against several drug resistant strains of M . tuberculosis.

Scand J Gastroenterol, 2001 Jun, 36(6), 584 - 8
Clarithromycin-based triple therapy for non-resistant Helicobacter pylori infection . How long should it be given?
Ogura K, Yoshida H, Maeda S, Yamaji Y, Kawabe T, Okamoto M, Shiratori Y, Omata M.
BACKGROUND: We have previously reported mixed infection with wild-type (sensitive) and mutant (resistant) Helicobacter pylori strains using a PCR-based preferential homo-duplex formation assay (PCR-PHFA) to detect gene mutations associated with clarithromycin resistance . Half of the cases with mixed infection were determined as sensitive by conventional MIC assessment and yet failed to respond to clarithromycin-based therapy . The aim of this study is to assess the efficacy of clarithromycin-based triple therapy in patients infected exclusively with wild-type strains as determined by PCR-PHFA . METHODS: Ninety patients who had pure wild-type H . pylori infection were randomly assigned to receive clarithromycin (200 mg b.i.d.), amoxicillin (500 mg q.i.d.) and lansoprazole (30 mg b.i.d.) for either 5 days or 7 days (n = 48 and n = 42, respectively) . The outcome of eradication was assessed by {13C} urea breath test . RESULTS: Eradication was achieved in 36/48 (75%) versus 39/42 (93%) by intention-to-treat analysis (P = 0.02), and in 36/45 (80%) versus 39/40 (98%) by per protocol analysis (P = 0.01), for the 5-day and 7-day protocols, respectively . Compliance and the incidence of untoward effects were similar in both groups . CONCLUSIONS: Seven-day administration is necessary and sufficient for the triple therapy with clarithromycin, amoxicillin and lansoprazole in patients with pure wild-type H . pylori infection.

Helicobacter, 2001 Jun, 6(2), 125 - 9
Inhibitory action of a novel proton pump inhibitor, rabeprazole, and its thioether derivative against the growth and motility of clarithromycin-resistant Helicobacter pylori; Ohara T et al.; BACKGROUND: Clarithromycin-resistant Helicobacter pylori (CRHP) has increasingly been isolated from patients in Japan . The aim of our study was to test whether proton pump inhibitors (PPIs) and their thioether derivatives, which are secreted into the gastric mucosa, could inhibit the growth and motility (a factor in colonization) of CRHP . MATERIALS AND METHODS: CRHP was isolated from patients who had experienced gastritis or peptic ulcers in Tokyo and Niigata . Drugs and related agents tested were omeprazole, lansoprazole, rabeprazole, the thioether derivative of rabeprazole (rabeprazole-TH), clarithromycin, amoxicillin and metronidazole . The MICs of the drugs and agents for H . pylori strains were determined by the agar dilution METHOD: Bacterial swimming in a liquid layer was examined under an inverted, phase-contrast microscope . RESULTS: The PPIs and rabeprazole-TH, but not the anti-H . pylori agents, inhibited the motility of CRHP at both pH 7.4 and 6.0 . The concentrations (microg/ml) necessary to inhibit 50% of the motility at pH 7.4 were 0.25-0.5, 8-32, 8-16 and 128-256 for rabeprazole-TH, rabeprazole, lansoprazole and omeprazole, respectively . Rabeprazole-TH exhibited the strongest inhibitory effect against the growth of CRPH (MIC, 0.5 microg/ml) . CONCLUSION: Rabeprazole-TH, which is secreted into the gastric mucosa, had the strongest inhibitory action against both the growth and motility of CRHP, suggesting that it is a potential novel agent for CRHP eradication.

J Ind Microbiol Biotechnol, 2001 Mar, 26(3), 171 - 7
Effects of acetic acid and lactic acid on the growth of Saccharomyces cerevisiae in a minimal medium; Narendranath NV et al.; Specific growth rates (mu) of two strains of Saccharomyces cerevisiae decreased exponentially (R2 > 0.9) as the concentrations of acetic acid or lactic acid were increased in minimal media at 30 degrees C . Moreover, the length of the lag phase of each growth curve (h) increased exponentially as increasing concentrations of acetic or lactic acid were added to the media . The minimum inhibitory concentration (MIC) of acetic acid for yeast growth was 0.6% w/v (100 mM) and that of lactic acid was 2.5% w/v (278 mM) for both strains of yeast . However, acetic acid at concentrations as low as 0.05-0.1% w/v and lactic acid at concentrations of 0.2-0.8% w/v begin to stress the yeasts as seen by reduced growth rates and decreased rates of glucose consumption and ethanol production as the concentration of acetic or lactic acid in the media was raised . In the presence of increasing acetic acid, all the glucose in the medium was eventually consumed even though the rates of consumption differed . However, this was not observed in the presence of increasing lactic acid where glucose consumption was extremely protracted even at a concentration of 0.6% w/v (66 mM) . A response surface central composite design was used to evaluate the interaction between acetic and lactic acids on the specific growth rate of both yeast strains at 30 degrees C . The data were analysed using the General Linear Models (GLM) procedure . From the analysis, the interaction between acetic acid and lactic acid was statistically significant (P < or = 0.001), i.e., the inhibitory effect of the two acids present together in a medium is highly synergistic.

J Antimicrob Chemother, 2001 Jul, 48 Suppl 1, 43 - 57
BSAC standardized disc susceptibility testing method; Andrews JM; BSAC Working Party On Susceptibility Testing ft; For nearly a decade microbiologists have used the MIC breakpoints published in the BSAC Guide to Susceptibility Testing to interpret susceptibility . Historically, and unlike the rest of Europe, the UK and Ireland have used a comparative method of disc testing to interpret susceptibility rather than one based on a correlation between MIC and zone of inhibition . Although innovative when introduced in the 1970s, Stokes' comparative method has evolved ad hoc and it has become increasingly apparent that there is a need for a standardized method of disc testing that is correlated with BSAC MIC breakpoints . The method described here, like all other standardized methods of disc testing, cannot be adapted by the user, and interpretative criteria are only applicable if the method is adhered to fully . A major advantage of this approach to susceptibility testing is that data from several sources can be combined for surveillance of resistance, a task that has been made much easier by the introduction of this method and coincides with the availability of automated zone measuring devices . It is hoped that the method described here will provide the core document for standard operating procedures; however, changes will necessarily occur over time as the method is developed and refined.

J Antimicrob Chemother, 2001 Jul, 48(1), 75 - 81
Resistance to itraconazole in Aspergillus nidulans and Aspergillus fumigatus is conferred by extra copies of the A . nidulans P-450 14alpha-demethylase gene, pdmA; Osherov N et al.; Triazoles selectively inhibit the cytochrome P-450-dependent C-14 lanosterol alpha-demethylase (P-450 14 alpha DM), a key enzyme in ergosterol biosynthesis in fungi . To investigate mechanisms of triazole resistance in a mould, we used Aspergillus nidulans, a genetically amenable model fungus closely related to more pathogenic members of the genus . We selected for genes that would give resistance to itraconazole following transformation with a high copy genomic library of A . nidulans . In all the resistant colonies that we isolated, resistance was conferred by extra copies of the A . nidulans P-450 14 alpha DM gene, pdmA . We determined that in A . nidulans, extra copies of pdmA increase the MIC for itraconazole 36 times over wild-type controls . Similarly, transformation of an Aspergillus fumigatus strain with pITZR1 resulted in increased resistance to itraconazole . Our results indicate that triazole resistance in clinical isolates of moulds may result from amplification or overexpression of the P-450 14 alpha DM and demonstrate the utility of A . nidulans as a promising model fungus for the analysis of drug resistance and susceptibility in the pathogenic fungus A . fumigatus.

Am J Perinatol, 2001 May, 18(3), 141 - 6
Antifungal susceptibility testing and the correlation with clinical outcome in neonatal candidemia; Huang YC et al.; The objective of this article is to assess the distribution of minimal inhibition concentrations (MIC) for candidal isolates from bloodstreams in neonates and to assess the correlation of clinical outcome with antifungal susceptibility testing . Of the 62 episodes of neonatal candidemia in a Children's Hospital between January 1994 and July 1998, 38 stocked isolates from 38 infants' bloodstreams were available and underwent antifungal susceptibility test according to National Committee for Clinical Laboratory Standards M27-A document . Correlation of clinical response with in vitro results was assessed in 37 patient-episode-isolate events . No less than 90% of these isolates tested were susceptible to amphotericin B, flucytosin, and fluconazole . The ranges of amphotericin B MICs and flucytosin MICs were narrow, ranging from 0.25 to 2 microg/mL, respectively . The range of fluconazole MICs was broad, ranging from 0.25 to >64 microg/mL . Successful therapy was achieved in 18 (62%) of 29 amphotericin B-treated patient-episode-susceptible isolate (MIC < or =1 microg/mL) events and 9 (64%) of 14 fluconazole-treated patient-episode-susceptible isolate events, respectively . Most isolates from the bloodstreams of neonates with candidemia were susceptible to antifungal agents tested but a low MIC of the antifungal agent did not predict successful therapy in this study . Correlating MICs with clinical outcome in neonatal candidemia requires complex evaluation of other factors.

Southeast Asian J Trop Med Public Health, 2000, 31 Suppl 1, 178 - 82
Antifungal activity of Curcuma longa grown in Thailand; Wuthi-udomlert M et al.; Curcuma longa Linn . or turmeric (Zingiberaceae) is a medicinal plant widely used and cultivated in tropical regions . According to Thai traditional texts, fresh and dried rhizomes are used as peptic ulcer treatment, carminatives, wound treatment and anti-inflammatory agent . Using hydro distillation, 1.88% and 7.02% (v/w) volatile oils were extracted from fresh and dried rhizomes, respectively, and 6.95% (w/w)crude curcuminoids were extracted from dried rhizomes . Dried powder was extracted with 95% ethanol and yielded 29.52% (w/w) crude ethanol extract composed of curcumin (11.6%), demethoxycurcumin (10.32%) and bisdemethoxycurcumin (10.77%) . These extracts were tested for antifungal activity by agar disc diffusion method against 29 clinical strains of dermatophytes . It was found that crude ethanol extract exhibited an inhibition zone range of 6.1 to 26.0 mm . There was no inhibition activity from crude curcuminoids while curcumin, demethoxycurcumin and bisdemethoxycutcumin gave different inhibition zone diameters ranging from 6.1 to 16.0 mm . Although antifungal activity of undiluted freshly distilled oil and 18-month-old oil revealed some differences, the inhibition zone diameters for both extracts varied within 26.1 to 46.0 mm . With 200 mg/ml ketoconazole, the activities of the standard agent were similar to the oil, both freshly distilled and 18-month-old, but were significantly different from those of curcuminoid compounds and crude ethanol extracts (p < 0.01) . Turmeric oil was also tested for its minimum inhibitory concentration (MIC) by broth dilution method . The MICs of freshly distilled and 18-month-old oils were 7.8 and 7.2 mg/ml respectively.

Mycoses, 2001 May, 44(3-4), 99 - 107
In-vitro and in-vivo anti-Trichophyton activity of essential oils by vapour contact; Inouye S et al.; The minimum inhibitory doses (MIDs) of essential oils by vapour contact to inhibit the growth of Trichophyton mentagrophytes and Trichophyton rubrum on agar medium were determined using airtight boxes . Among seven essential oils examined, cinnamon bark oil showed the least MID, followed by lemongrass, thyme and perilla oils . Lavender and tea tree oils showed moderate MID, and citron oil showed the highest MID, being 320 times higher than that of cinnamon bark oil . The MID values were less than the minimum inhibitory concentration (MIC) values determined by agar dilution assay . Furthermore, the minimum agar concentration (MAC) of essential oils absorbed from vapour was determined at the time of MID determination as the second antifungal measure . The MAC value by vapour contact was 1.4 to 4.7 times less than the MAC remaining in the agar at the time of MIC determination by agar dilution assay . Using selected essential oils, the anti-Trichophyton activity by vapour contact was examined in more detail . Lemongrass, thyme and perilla oils killed the conidia, inhibited germination and hyphal elongation at 1-4 micrograms ml-1 air, whereas lavender oil was effective at 40-160 micrograms ml-1 air . The in-vivo efficacy of thyme and perilla oils by vapour contact was shown against an experimental tinea pedis in guinea pigs infected with T . mentagrophytes . These results indicated potent anti-Trichophyton action of essential oils by vapour contact.

Mycoses, 2001 May, 44(3-4), 109 - 12
Antifungal activities of pelargonic and capric acid on Microsporum gypseum; Chadeganipour M et al.; The inhibitory effects of pelargonic and capric acid on Microsporum gypseum were examined . Solid and liquid Sabouraud glucose media containing different concentrations of pelargonic and capric acid were separately prepared and inoculated with the suspension of mycelium and spores of M . gypseum and incubated at 25 degrees C for 1 month . The culture media were examined periodically for fungal growth and the minimum inhibitory concentration (MIC) of each fatty acid was determined . The MIC for capric acid was 0.02 mg ml-1 and for pelargonic acid 0.04 mg ml-1 on solid media and 0.075 mg ml-1 for capric acid and 0.05 mg ml-1 for pelargonic acid in the liquid media.

Arch Pharm (Weinheim), 2001 May, 334(5), 143 - 7
Antifungal actinomycete metabolites discovered in a differential cell-based screening using a recombinant TOPO1 deletion mutant strain; Stadler M et al.; In the course of a natural product screening for inhibitors of fungal topoisomerase 1 (TOPO 1), extracts from the actinomycete strains WS 1410 and BS 1465 exhibited promising activities . Bioguided fractionation of the culture broth by preparative HPLC methods yielded the collismycins A (1) and B (2) as active principles of strain WS 1410 . Out of the mycelial extracts of strain BS 1465 the bioactive new natural products, cyclo-homononactic acid (3) and cyclo-nonactic acid (5) and the structurally related but inactive homononactic acid (4), were isolated . Both collismycin isomers inhibited the recombinant yeast strains ScAL 141 and ScAL 143 (TOPO 1 deletion mutant) in a non-specific manner with an MIC in the range of 2 micrograms/ml . The novel cyclo-homononactic acid (3) and cyclo-nonactic acid (5) showed higher selectivity towards the wild type strain (MIC = 2 micrograms/ml as compared to 10 micrograms/ml for the deletion mutant) . All compounds obviously address a target other than TOPO 1 since they do not exhibit activities in a concurrent TOPO 1 enzyme assay.

Int J Tuberc Lung Dis, 2001 Jun, 5(6), 533 - 8
The early bactericidal activity of amikacin in pulmonary tuberculosis; Donald PR et al.; SETTING: Stellenbosch University, a tertiary care hospital in Cape Town, South Africa . OBJECTIVE: To determine the early bactericidal activity (EBA) of amikacin in dosages of 5 mg/kg, 10 mg/kg and 15 mg/kg body weight in comparison to that of isoniazid 6 mg/kg body weight or no drug . DESIGN: An open, randomised trial . PATIENTS: Patients with previously untreated, sputum smear-positive pulmonary tuberculosis . INTERVENTION: Patients received amikacin 5 mg/kg (12 patients), 10 mg/kg (13 patients) or 15 mg/kg (15 patients), isoniazid 6 mg/kg (9 patients) or no drug (10 patients) . RESULTS: The rate of decrease in log viable colony forming units of Mycobacterium tuberculosis per ml of sputum per day during the first 2 days of treatment with amikacin 5 mg/kg, 10 mg/kg and 15 mg/kg was 0.041 (SD 0.100), 0.045 (SD 0.144) and 0.052 (SD 0.096), respectively, 0.515 (SD 0.173) in the patients receiving isoniazid 6 mg/kg, and 0.041 (SD 0.113) in those receiving no drug . The EBA found in patients receiving amikacin did not differ significantly from that of the no drug group . However, as the EBA in the no drug group was the highest ever encountered at Stellenbosch University, the mean in patients receiving drug was tested against 0 and found to differ significantly (P = 0.03), suggesting minimal activity . Mean amikacin serum concentrations 1 hour after intramuscular drug administration were 13.5 microg/ml, 26.7 microg/ml and 39.2 microg/ml in the patients receiving 5 mg, 10 mg and 15 mg per kg body weight, respectively . CONCLUSION: Despite serum concentrations well in excess of the minimal inhibitory concentration of 2-4 microg/ml, the EBA of amikacin in patients with smear-positive pulmonary tuberculosis was only just detectable.

Cornea, 2001 Jul, 20(5), 458 - 62
Treatment and outcome of nocardia keratitis; Sridhar MS et al.; OBJECTIVE: To report our experience in treatment and outcome ofNocardia keratitis . METHODS: Medical and microbiology records of seven cases of culture-provenNocardia keratitis seen between January 1997 and March 1999 were reviewed retrospectively . In all patients, corneal scrapings were obtained for direct microscopic evaluation and culture . Drug sensitivity was determined by the Kirby-Bauer disk-diffusion method . The minimum inhibitory concentration of ciprofloxacin and amikacin for these isolates was determined by agar dilution method . Response to medical therapy and the end result were analyzed . RESULTS: By the in vitro Kirby-Bauer disk-diffusion techniques, all isolates were sensitive to amikacin; six of these isolates were sensitive to gentamicin and four were sensitive to ciprofloxacin . The minimum inhibitory concentration (MIC) of amikacin for all isolates by the agar-dilution technique was well below the MIC breakpoint forNocardia resistance, whereas the MIC of ciprofloxacin was above the MIC breakpoint forNocardia resistance . All patients responded to medical therapy . The corneal infection resolved in three patients after treatment with ciprofloxacin, in one patient after fortified gentamicin, and in three patients after fortified amikacin . Outcome details were available for six patients . There was good visual recovery in four patients, with visual acuity of 20/25 or better in three . The cornea of two patients developed nonvascularized scars, and in four patients in whom the infiltrates were peripheral, vascularization was seen . CONCLUSIONS: Although patients ofNocardia keratitis may respond to other antibiotics, amikacin appears to be a drug of choice . In this small series, when appropriate therapy was initiated,Nocardia keratitis resolved promptly with good visual recovery.

Antimicrob Agents Chemother, 2001 Jul, 45(7), 2070 - 4
Detection of resistance to amphotericin B in Candida isolates by using Iso-Sensitest broth; Cuenca-Estrella M et al.; A major limitation of the National Committee for Clinical Laboratory Standards M27-A methodology is reliable detection of amphotericin B (AMB) resistance . The results obtained by using Iso-Sensitest, a synthetic medium, to detect AMB resistance were analyzed and compared with those obtained with RPMI and antibiotic medium 3 (AM3) . The ability to detect AMB resistance with RPMI is not enhanced by using a higher inoculum, glucose supplementation at a final concentration of 20 g/liter, spectrophotometric reading, or 24 h of incubation time . Testing using AM3 and an inoculum of 10(3) CFU/ml detects resistance . Identification of resistant isolates is not improved by glucose supplementation, changes in reading method, or changes in incubation time . However, the use of AM3 as assay medium and an inoculum of 10(5) CFU/ml did not allow detection of AMB resistance . Testing using Iso-Sensitest medium appears to be similar to AM3 in detecting resistance . The most pronounced discrimination is achieved by testing in Iso-Sensitest supplemented with glucose and spectrophotometric reading after 24 h of incubation . The reproducibility of MIC testing was greatest for Iso-Sensitest-based procedures . Use of Iso-Sensitest produces both highly reproducible MICs and reliable identification of AMB-resistant Candida isolates.

Pest Manag Sci, 2001 Jun, 57(6), 554 - 9
Activity against plant pathogenic fungi of phomalactone isolated from Nigrospora sphaerica; Kim JC et al.; Phomalactone, 5,6-dihydro-5-hydroxy-6-prop-2-enyl-2H-pyran-2-one, produced by the fungus Nigrospora sphaerica, was tested in vitro against nine plant pathogenic fungi, and specifically inhibited the mycelial growth of Phytophthora infestans, with an MIC value of 2.5 mg litre-1 . Its inhibitory activities against sporangium and zoospore germination of P infestans were similar to those against Phytophthora capsici . In vivo, at 100 and 500 mg litre-1, it reduced the development of tomato late blight caused by P infestans.

J Personal Disord, 2001 Jun, 15(3), 255 - 62
Applicability of personality disorder criteria in late adolescence: internal consistency and criterion overlap 2 years after psychiatric hospitalization; Becker DF et al.; We examined internal consistency and criterion overlap of DSM-III-R personality disorder criteria in late adolescence, 2 years after psychiatric hospitalization . A total of 60 adolescents were assessed with the Personality Disorder Examination . Within-category cohesiveness (internal consistency) was evaluated by coefficient alpha and mean intercriterion correlation (MIC) . Between-category criterion overlap was evaluated by examining intercategory mean intercriterion correlations (ICMIC) between all pairs of disorders . Internal consistency was low, with alpha less than .70 for all except borderline and dependent personality disorders . For most disorders, MIC values were higher than ICMIC values . Our findings suggest that personality disorder criteria sets have limited internal consistency in older adolescents . Although the criteria for most personality disorders correlate better with each other than with the criteria for other personality disorders, suggesting some degree of discriminant validity, comparison of these results with a similar analysis performed shortly after hospital admission raises questions about personality disorder construct validity during adolescence.

Ophthalmologica, 2001 Jul-Aug, 215(4), 299 - 302
Penetration of amikacin into aqueous humor of rabbits; Erkin EF et al.; Amikacin is an aminoglycoside antibiotic that has poor corneal penetration due to its hydrophilic properties . The purpose of this study was to compare and evaluate the penetration of amikacin sulfate into aqueous humor of the rabbit eye when applied by different routes and concentrations, namely 100 or 250 mg/ml topical fortified amikacin eye drops, 100 or 250 mg/ml amikacin-embedded soft contact lenses and 25 mg subconjunctival amikacin injection . One hour after application, amikacin was not detectable in any of the 100 mg/ml concentration groups . High levels of amikacin above the minimum inhibitory concentration for susceptible bacteria were detected when applied subconjunctivally and by 250 mg/ml topical fortified routes . Topical fortified amikacin 250 mg/ml reached the highest value in the aqueous (p < 0.05) . Our results point out the poor corneal penetration of amikacin in standard concentrations from the intact rabbit cornea and that subconjunctival injections might provide satisfactory penetration .

Rinsho Byori, 2001 May, 49(5), 505 - 11
{An approach for the quality control in clinical microbiology laboratory by antibiotic susceptibility}; Ueno T et al.; It is important to develop a system for the prevention of various errors and the maintenance of the high quality level in the department of laboratory medicine . We developed a computer program to infer the species of microorganisms by assessing the minimal inhibitory concentration of respective microorganisms to various antibiotics, antibiotic susceptibility patterns(ASP) . Retrospective analysis of 3,405 strains from 39 species using this program resulted in a sensitivity of 91.4% and specificity of 98.9% . Furthermore, in 1,660 strains isolated from September 1999 to August 2000, the efficacy of this program was evaluated using both this program and conventional identifying method such as VITEK, api and ATB . The discrepancy between this inferring method and conventional method was noted in 179 strains, revealing errors in 12 strains . We conclude that this ASP inferring program is useful for preventing errors and the maintenance of high quality examination by combining with conventional methods in the laboratory.

An Med Interna, 2001 Jan, 18(1), 35 - 7
{Addisonian crisis as first manifestation of adrenal gland insufficiency in patient diagnosed with lung cancer}; Munoz A et al.; Adrenal insufficiency or Addison's disease is actually a rare illness associated with numerous pathologies . We describe the case of a fifty years old male with lung adenocarcinoma and metastasis in both adrenal glands, who was receiving chemotherapy with mytomicin, ifosfamide and cisplatin (MIC), and was diagnosed of adrenal insufficiency as a result of acute episode addisonian crisis . Many times, the clinic symptoms of adrenal insufficiency can go unnoticed due to its low specifity and to mixing up with other syndromes . Hypoadrenalism has been described in association with many tumours, specially with non-Hodgkin's lymphoma . It seems that there is a discordance between the number of patients with bilateral metastatic adrenal destruction and the documented cases of clinic insufficiency . Once the adrenal failure is suspected, the diagnosis and hormone replacement treatment are really easy . Addison's disease ethiologies are revised putting special emphasis on those related with cancer patients.

Eur J Pharm Sci, 2001 Jun, 13(3), 243 - 8
In vitro evaluation of 2,4-dihydroxythiobenzanilides against various moulds; Niewiadomy A et al.; The antimycotic potency of 2,4-dihydroxythiobenzanilide derivatives was tested . The MIC assessments by an agar dilution method were used for the estimation of potential activity in vitro against the four mould strains: Scopulariopsis brevicalis, Aspergillus niger, Aspergillus fumigatus and Penicillium sp . The strongest fungistatic activity was observed for 3'-fluoro-derivative (MIC 7.82 microg/ml) . It was stated that the inhibition action of these compounds depends mainly on lipophilicity of molecules . Parabolic relationships between the antimycotic activity and lipophilicity were found.

Int J Pharm, 2001 Jun 4, 220(1-2), 169 - 77
Relevance of polymer molecular weight to the in vitro/in vivo performances of ocular inserts based on poly(ethylene oxide); Di Colo G et al.; A previous study of the present authors on gel-forming erodible inserts, based on high molecular weight (MW, 400 kDa) poly(ethylene oxide) (PEO), for ocular controlled delivery of ofloxacin (OFX) has been extended to investigate the effects of PEO MW, in the 200-2000 kDa range, on insert properties relevant to therapeutic efficacy . Mucoadhesion has shown a dependence on MW, with a maximum for PEO 400 . The in vitro drug release from inserts based on PEO 200, PEO 400 and PEO 900 was mainly controlled by insert erosion, whereas with PEO 2000 it was mainly diffusion-controlled in a first phase, followed by an erosion-controlled phase . The erosion time scale depended directly on MW . Immediately after application in the lower conjunctival sac of the rabbit eye, the inserts based on PEO of whichever MW formed mucoadhesive gels, well tolerated by the animals; then the gels spread over the corneal surface and eroded . PEO 2000 was unsuitable as an insert material, since the resulting gel spilled from the eye, due to excessive swelling . The gel residence time in the precorneal area, the drug permanence time in the aqueous humor at concentrations > MIC and the time to reach the maximal drug concentration in the aqueous humor (C(max)) depended directly on MW, indicating that transcorneal absorption was governed by gel erosion . All inserts increased Cmax and AUCeff (AUC for concentrations > MIC) with respect to the commercial eyedrops . The increases caused by PEO 400 and PEO 900 were similar (3.78- and 3.16-fold, respectively, for Cmax; 11.06- and 12.37-fold, respectively, for AUCeff), whereas smaller increases were produced by PEO 200 . The PEO 400 and PEO 900 inserts have shown a potential for a topical treatment of endophthalmitis.

Rev Esp Quimioter, 2001 Mar, 14(1), 40 - 6
{Comparative study of 3 tests for susceptibility of Mycobacterium tuberculosis to first-line antituberculous drugs}; Delas MA et al.; Treatment of tuberculosis requires a multidrug regimen for a prolonged period of time (6-12 months), which leads to a high level of noncompliance with therapy . This lack of compliance has led to an increase in drug-resistant strains of Mycobacterium tuberculosis . Rapid drug susceptibility tests are needed to assess effective chemotherapy regimens . In the present study, the reliability of ESP Myco System II(R) and E-test(R) for determining the susceptibility of 82 clinical isolates of M . tuberculosis to rifampin, isoniazid, streptomycin and ethambutol was evaluated by comparing results to those of the multiple proportion method as a reference . ESP Myco System II(R) is a nonradioactive, totally automated, continuously monitored system designed to detect grown mycobacteria . E-test(R) is a well-described method of susceptibility testing which provides data on minimal inhibitory concentration . The susceptibility results of both methods were available within 9 days, while those of the proportion method required 28 days . Levels of agreement between ESP Myco System II(R) and the proportion method for the clinical isolates analyzed were 100% for rifampin, isoniazid and ethambutol . Only one discrepancy was found for streptomycin . For the E-test(R), the results were less favorable . ESP Myco System II(R) was shown to be a rapid and reliable method for testing susceptibility of M . tuberculosis isolates . Nevertheless, E-test(R) provided low specificity for streptomycin and ethambutol . Further studies to test the reliability of the E-test(R) are necessary.

J Nat Prod, 2001 May, 64(5), 559 - 62
Antimalarial, cytotoxic, and antifungal alkaloids from Duguetia hadrantha; Muhammad I et al.; Bioassay-guided isolation of Duguetia hadrantha yielded two new 4,5-dioxo-1-azaaporphinoids, hadranthine A (1) and hadranthine B (2), together with the known alkaloids imbiline-1 (3), sampangine (4), and 3-methoxysampangine (5), whose structures were determined primarily from 2D-NMR 1H-13C HMBC, and 1H-15N HMBC experiments . This is the first report of the co-occurrence of the copyrine alkaloids 4 and 5, as well as the first report of either copyrine or imbiline type alkaloids from a Duguetia species . Compounds 1, 4, and 5 demonstrated in vitro antimalarial activity against Plasmodium falciparum (W-2 clone), while 2 was inactive . Instead, 2 showed in vitro cytotoxicity to selected human cancer cell lines (IC50 = 3-6 microg/mL against SK-MEL, KB, BT-549, and SK-OV-3), and 4 was also cytotoxic to human malignant melanoma (IC50 = 0.37 microg/mL) . Sampangine (4) also inhibited cell aggregation with a MIC value of <0.15 microg/mL, while 3-methoxysampangine (5) was only weakly active.

Med Biol Eng Comput, 2001 Mar, 39(2), 195 - 201
Mechanical behaviour of condenser microphone in mechanomyography; Watakabe M et al.; Condenser microphones (MIC) have been widely used in mechanomyography, together with accelerometers and piezoelectric contact sensors . The aim of the present investigation was to clarify the mechanical variable (acceleration, velocity or displacement) indicated by the signal from a MIC transducer using a mechanical sinusoidal vibration system . In addition, the mechanomyogram (MMG) was recorded simultaneously with a MIC transducer and accelerometer (ACC) during voluntary contractions to confirm the mechanical variable reflected by the actual MMG and to examine the influence of motion artifact on the MMG . To measure the displacement-frequency response, mechanical sinusoidal vibrations of 3 to 300 Hz were applied to the MIC transducer with different sizes of air chambers (5, 10, 15 and 20 mm in diameter and 15, 20 or 25 mm long) . The MIC transducer showed a linear relationship between the output amplitude and the vibration displacement, however, its frequency response declined with decreasing diameter and decreasing length of the air chamber . In fact, the cut-off frequency (-3dB) of the MIC transducer with the 5-mm-diameter chamber was 10, 8 and 4 Hz for the length 15, 20 and 25 mm, respectively . The air chamber with at least a diameter of 10 mm and a length of 15 mm is recommended for the MIC transducer . The sensitivity of this MIC transducer arrangement was 92 mV microm(-1) when excited at 100 Hz . During voluntary contraction, the amplitude spectral density function of the MMG from the MIC transducer resembled that of the double integral of the ACC transducer signal . The angle of the MIC transducer was delayed by 180 degrees in relation to the ACC transducer signal . The sensitivity of the MIC transducer was reduced to one-third because of the peculiar volume change of air chamber when the MMG was detected on the surface of the skin . In addition, the MIC transducer was contaminated by a smaller motion artifact than that from the ACC transducer . The maximal peak amplitude of the MIC and ACC transducer signal with the motion artifact was 7.7 and 12.3 times as much as the RMS amplitude of each signal without the motion artifact, respectively . These findings suggest that the MIC transducer acts as a displacement meter in the MMG . The MIC transducer seems to be a possible candidate for recording the MMG during dynamic muscle contractions as well as during sustained contractions.

Chirurg, 2001 Apr, 72(4), 362 - 7
{Minimally invasive surgery (MIC) in the area of the colon and rectum: technique of minimally invasive colorectal surgery}; Szinicz G et al.; Minimally invasive surgery is gaining increased importance in colorectal surgery . Since 1993 we have performed 378 minimally invasive colorectal procedures (appendzcetomy excluded): right-sided (n = 70) and left-sided (n = 160) hemicolectomy, anterior rectal resections (n = 72), and colostomy (n = 12), Hartmann procedure (n = 17), take-down operation (n = 25), rectopexy (n = 21) and coloproctomucosectomy (n = 2) . The indications were diverticulitis (n = 147), benign tumor (n = 40), malignant tumor (n = 135), rectal prolapse (n = 21), Crohn's disease (n = 8), intestinal diversion (n = 25), colonic perforation (n = 2) . Postoperative complications were observed in 21% of cases; anastomotic leak developed in 4%; mortality was 0.5% . In keeping with the current literature our data show that in the majority of cases surgical therapy of colorectal disease can be performed in minimally invasive fashion.

Vet Microbiol, 2001 Jul 3, 81(1), 79 - 84
Activity and postantibiotic effect of marbofloxacin, enrofloxacin, difloxacin and ciprofloxacin against feline Bordetella bronchiseptica isolates; Carbone M et al.; The minimum inhibitory concentration (MIC) and postantibiotic effect (PAE) of marbofloxacin, enrofloxacin, difloxacin and ciprofloxacin were evaluated in vitro against 43 feline-source Bordetella bronchiseptica strains . All strains tested were susceptible to marbofloxacin and enrofloxacin (MIC90 0.5mg/l), while 93 and 84% of the strains were susceptible, respectively, to ciprofloxacin and difloxacin with MIC(90) values of, respectively, 1 and 8mg/l . The PAE was studied in 10 strains by exposure of bacteria to marbofloxacin, enrofloxacin, difloxacin and ciprofloxacin at 5 and 10 times minimum inhibitory concentration (MIC) for 1 and 2h . Regrowth was determined by measuring the viable counts after drug removal by a 10(3) dilution procedure . PAEs increased as a function of concentration and exposure time . The mean duration of PAEs varied between 1.1 and 8.2h, showing the following order: marbofloxacin>enrofloxacin>ciprofloxacin>difloxacin . These data are encouraging since fluoroquinolones have a possible role in the clinical treatment of B . bronchiseptica infections, and the strong PAE caused by quinolones may contribute to the in vivo efficacy of these drugs.

Parasitol Res, 2001 Apr, 87(4), 281 - 6
In vitro taurocholate-induced segmentation and clustering of Mesocestoides vogae (syn . corti) tetrathyridia (Cestoda)--inhibition byh cestocidal drugs; Saldana J et al.; Mesocestoides vogae (syn . M . corti) tetrathyridia were cultured in the presence of sodium taurocholate, for the purpose of exploring the suitability of this organism for the in vitro assay of cestocidal drugs . Parasite clustering and segmentation were observed as taurocholate-dependent effects in biphasic and monophasic media, respectively . Interestingly, representative members of two major classes of known cestocidal agents (namely, albendazole and praziquantel) blocked these effects . Furthermore, it was possible to determine a specific concentration of the drugs that inhibited clustering and segmentation (minimum inhibitory concentration) . In contrast, no inhibition was obtained in the presence of anthelmintics without cestocidal activity . These observations open the way for further studies focused at understanding how the activity of the drugs is involved in the suppression of the taurocholate-induced effects.

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