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