Minimal inhibiting concentration

Bone Marrow Transplant . 2005 Jan 17; {Epub ahead of print}
Voriconazole therapeutic drug monitoring in allogeneic hematopoietic stem cell transplant recipients; Trifilio S et al.; Summary:Voriconazole, a new antifungal agent, is increasingly being used after HSCT . The hepatic cytochrome P450 isoenzyme 2C19 plays a significant role in voriconazole metabolism . As CYP2C19 exhibits significant genetic polymorphism, some patients metabolize voriconazole poorly resulting in increased plasma drug levels . The clinical significance of this is unknown, and the utility of monitoring voriconazole levels is unclear . Steady-state trough plasma voriconazole levels were obtained in 25 allogeneic HSCT recipients using an HPLC assay . Patients had drug levels checked once (n=13), twice (n=10), or >/=3 times (n=2) 5-18 days (median 10) after starting voriconazole or dose modification . The 41 voriconazole levels were 0.2-6.8 mug/ml (median 1.6); 6 (15%) were <0.5 (possibly below the in vitro MIC(90) for Aspergillus spp.) . Voriconazole concentrations correlated with aspartate aminotranferase (AST) (r=0.5; P=0.0009) and alkaline phosphatase (r=0.34; P=0.03), but not with creatinine, bilirubin and alanine aminotransferase (ALT) . Since liver dysfunction is common after HSCT, it was not possible to determine if elevated AST and alkaline phosphatase levels were the cause or the consequence of higher voriconazole levels . We conclude that trough voriconazole levels vary considerably between patients, and suggest monitoring levels in patients receiving voriconazole for confirmed fungal infections, and in those with elevated AST or alkaline phosphatase levels.Bone Marrow Transplantation advance online publication, 17 January 2005; doi:10.1038/sj.bmt.1704828.

Biosens Bioelectron, 2004 Dec 15, 20(6), 1068 - 75
A molecularly imprinted catalyst designed by a computational approach in catalysing a transesterification process; Meng Z et al.; A computational approach was developed to optimize the monomer formulation of molecularly imprinted catalysts . A virtual library of the intermediates of a lipase-catalysed transesterification process was constructed using Chem3D software with p-nitrophenyl acetate as substrate . The energies of the intermediates were minimized using the semi-empirical MOPAC method with the most stable intermediate expected to lead to a higher turn over rate . According to the optimization results, a MIC was prepared by co-polymerising 4(5)-vinylimidazole and itaconic acid with trimethylpropanol trimethacrylate micro spheres in the presence of p-nitrophenyl acetate . The MIC achieved of the transesterification process between p-nitrophenyl acetate and hexanol with a turn over rate of 26.2 min(-1), and showed substrate specificity towards its template with a 6.5-fold preference for p-nitrophenyl acetate over p-nitrophenyl salicylate.

Drug Des Discov, 2003, 18(4), 103 - 8
In vitro and in vivo activities of acylated derivatives of isoniazid against mycobacterium tuberculosis; Hearn MJ et al.; Enzymatic acylation of the antitubercular isoniazid (INH) by N-acetyl transferases reduces the therapeutic effectiveness of the drug . Because it represents a major metabolic pathway for INH in human beings, such acetylation has serious consequences for tuberculosis treatment regimens . Among patients in whom this process is efficient, the "rapid acetylators," the resultant chronic underdosing of INH may give rise to the development of resistance, as well as inadequate therapy . Not much work has been done previously to characterize the antitubercular properties of other N2-acylisoniazids . In order to address the fundamental issue of the activities of these acylated derivatives of INH, a number of such compounds 1a-f were chemically synthesized for investigation by a method providing good yield and purity . In experiments in vitro against Mycobacterium tuberculosis, these compounds displayed minimum inhibitory concentration (MIC) values between several fold and several hundred fold greater than that of INH itself, on a molar basis, with some of the more active compounds having higher calculated values of log P . Among these derivatives, compound 1b, closely homologous to the INH metabolite 1a, N2-acetylisoniazid, provided unexpected protection in tuberculosis-infected mice . The authors conclude that such close structural congeners of metabolites of INH may serve as significant leads in antitubercular drug discovery and in the exploration of the mode of action of INH.

Planta Med, 2004 Nov, 70(11), 1093 - 5
Trypanocidal activity of a new diterpene from Casearia sylvestris var . lingua; Espindola LS et al.; Bioassay-guided fractionation of the hexanic root bark extract of Casearia sylvestris var . lingua led to the isolation of a new clerodane diterpene, whose structure was elucidated as rel-(2 S,5 R,6 R,8 S,9 S,10 R,18 S,19 R)-19-acetoxy-18,19-epoxy-6-hydroxy-18-butanoyloxy-2-(2-methylbutanoyloxy)cleroda-3,13(16), 14-triene by spectroscopic means, including 1D and 2D NMR analyses . This compound showed pronounced activity on Trypanosoma cruzi, the casual agent of Chagas' disease, with minimal inhibitory concentration (MIC) at 0.59 microg/mL.

Neoplasia, 2004 Sep-Oct, 6(5), 558 - 68
Downregulation and/or release of NKG2D ligands as immune evasion strategy of human neuroblastoma; Raffaghello L et al.; Neuroblastoma (NB) is a pediatric extracranial tumor characterized by downregulation of human leukocyte antigen class I and defects of the antigen processing machinery, two features that make it an appropriate target for natural killer (NK)-mediated lysis . NKG2D is an activating immunoreceptor expressed by cytotoxic T lymphocytes and NK cells . The ligands for NKG2D are the major histocompatibility complex class I-related chain (MIC)A and MICB glycoproteins, and the UL-16-binding proteins (ULBPs) . Here, the expression of NKG2D ligands was investigated in human primary NB tumors and cell lines because scanty information is available on this issue . MICA, MICB, and ULBP transcripts were found in most tumors and cell lines . MICA protein was detected in some NB cell lines but not in primary tumors . A soluble form of MICA (sMICA) was identified in most patient sera and in some cell line supernatants . sMICA downregulated surface NKG2D in normal peripheral blood CD8(+) cells and decreased NK-mediated killing of MICA(+) NB cells . MICB was detected exclusively in the cytosol of primary tumors and cell lines . Approximately 50% of primary tumors expressed ULBP-2, but not ULBP-1 or -3 . ULBP-3 was expressed in 5 of 9 cell lines, ULBP-2 in 2 of 9, whereas ULBP-1 was never detected . These studies delineate novel potential pathways of tumor escape and immunodeficiency in NB.

J Antimicrob Chemother, 2005 Jan, 55(1), 102 - 5 Epub 2004 Nov 16.
Determination of antifungal drug susceptibilities of Aspergillus species by a fluorescence-based microplate assay; Balajee SA et al.; OBJECTIVES: We have investigated the use of a viability dye, chloromethylfluorescein di-acetate (CMFDA), for antifungal susceptibility testing in a fluorescence microplate (FM) assay format . METHODS: For this FM assay, conidia were incubated in increasing concentrations of antifungal drug for 16 h and stained with CMFDA . Fluorescence, measured as mean fluorescence units (MFU) in a fluorescence microplate reader, was graphed relative to that of a drug-free control, and the MIC was defined as the lowest concentration of the drug that resulted in complete reduction (100%) in MFU for amphotericin B, or 90% reduction in MFU for itraconazole and voriconazole . Susceptibilities of 10 clinical isolates of Aspergillus fumigatus, Aspergillus terreus and Aspergillus niger to amphotericin B, itraconazole and voriconazole were tested in a blinded fashion using the FM and the NCCLS methods . RESULTS AND CONCLUSIONS: Reproducibility of the FM assay was excellent, and results correlated with those of the NCCLS microdilution method . The FM assay appears to be a rapid, objective method for testing fungal susceptibilities to itraconazole, voriconazole and amphotericin B.

J Antimicrob Chemother, 2005 Jan, 55(1), 106 - 109 Epub 2004 Nov 16.
Rapid flow-cytometric susceptibility testing of Candida species; Rudensky B et al.; OBJECTIVES: To develop a rapid flow-cytometric antifungal susceptibility test and to compare results with the standard methods . METHODS: Reference and laboratory strains of Candida were tested for susceptibility to fluconazole and echinocandin by fluorescent flow cytometry using Acridine Orange as indicator of viability . Flow cytometry results were compared with MICs as determined by macrodilution and/or Etest . RESULTS: Seventy Candida strains were tested for susceptibility to fluconazole, and 74 strains for susceptibility to echinocandin . Minimal concentration of fluconazole causing 40% cell damage, as determined by flow cytometry, showed excellent association with MIC, as determined by other methods . The flow method, completed within 5 h, had excellent sensitivity and specificity to distinguish between sensitive, susceptible dose-dependent and resistant strains . The flow cytometry method for echinocandin was completed within 3 h, and minimal concentration causing 50% cell damage was associated with MIC as determined by macrodilution . CONCLUSIONS: Antifungal susceptibility testing by FACS is a reliable, rapid method for determining susceptibility of Candida to fluconazole and echinocandin . The method allows same-day results, assisting in the selection of appropriate antifungal therapy.

J Antimicrob Chemother, 2005 Jan, 55(1), 78 - 83 Epub 2004 Nov 16.
Experimental study of teicoplanin, alone and in combination, in the therapy of cephalosporin-resistant pneumococcal meningitis; Fernandez A et al.; OBJECTIVES: The aim of the study was to determine the efficacy of teicoplanin, alone and in combination with ceftriaxone, in a rabbit model of cephalosporin-resistant pneumococcal meningitis, and to assess the effect of concomitant therapy with dexamethasone . METHODS: In vitro killing curves of teicoplanin, with and without ceftriaxone, were performed . Groups of eight animals per treatment were inoculated with a cephalosporin-resistant pneumococcal strain (penicillin MIC, 4 mg/L; ceftriaxone MIC, 2 mg/L; teicoplanin MIC, 0.03 mg/L) and treated over a 26 h period . Teicoplanin was administered at a dose of 15 mg/kg, alone and in combination with ceftriaxone at 100 mg/kg with or without dexamethasone at 0.25 mg/kg . CSF samples were collected at different time-points, and bacterial titres, white blood cell counts, lactate and protein concentrations and bacteriostatic/bactericidal titres were determined . Blood and CSF teicoplanin pharmacokinetic and pharmacodynamic parameters were determined . RESULTS: Teicoplanin alone promoted a decrease in bacterial counts at 6 h of -2.66 log cfu/mL and was bactericidal at 24 h, without therapeutic failures . Similar good results were obtained when dexamethasone was used simultaneously, in spite of the penetration of teicoplanin into the CSF being significantly reduced, from 2.31% to 0.71% . Teicoplanin and ceftriaxone combinations were synergic in vitro, but not in the meningitis model . CONCLUSIONS: Teicoplanin alone was very effective in this model of cephalosporin-resistant pneumococcal meningitis . The use of concomitant dexamethasone resulted in lower CSF teicoplanin levels, but not in therapeutic failures . The combination of teicoplanin plus ceftriaxone and dexamethasone might be a good alternative for the empirical therapy of pneumococcal meningitis . Additional data should confirm our experiments, in advance of clinical trials to assess efficacy in humans.

Acta Trop, 2004 Nov-Dec, 92(3), 237 - 44
In vitro activity of two phenyl-carbamate derivatives, singly and in combination with albendazole against albendazole-resistant Giardia intestinalis; Jimenez-Cardoso E et al.; Giardia intestinalis is one of the most prevalent parasites in adults and children in Mexico . Benzimidazoles have been proposed as a therapeutic alternative in the treatment of giardiasis . However, high-dose related toxicity and the development of resistance have emerged in clinical trials using this therapy . In the search of alternative drugs, we found that benzimidazole-resistant strains of fungi have shown increased sensitivity to phenyl-carbamates, hence, we developed several substituted phenyl-carbamates, two of which were tested against the protozoan parasite G . intestinalis in susceptible and albendazole-resistant Giardia strains . 4-R-ethyl-phenyl-carbamates IRE-6A and IRE-7B demonstrated antigiardial, albeit modest, activity when compared with albendazole, against susceptible and albendazole-induced resistant Giardia . However, when albendazole 0.38 microg/mL (MIC(50)) was combined with each IRE compound, a significant antigiardial synergism (fractional inhibitory concentration index (FICI < 0.5)) was obtained not only with sensitive cultures but also with resistant Giardia parasites . The results described here suggest a potential role for a combined therapy with phenyl-carbamates and sub-doses of benzimidazoles in the treatment of giardiasis.

J Zoo Wildl Med, 2004 Sep, 35(3), 341 - 6
Pharmacokinetic disposition of a long-acting oxytetracycline formulation after single-dose intravenous and intramuscular administrations in the American alligator (Alligator mississippiensis); Helmick KE et al.; The pharmacokinetics of a long-acting oxytetracycline preparation administered i.v . and i.m . to American alligators (Alligator mississippiensis) at 10 mg/kg was determined . Plasma levels of oxytetracycline were measured using high-performance liquid chromatography, and the resulting concentration versus time curve was analyzed using compartmental modeling and noncompartmental modeling techniques for i.v . and i.m . samples, respectively . A two-compartment model best represented the i.v . data . Intravenous administration of oxytetracycline resulted in an extrapolated mean plasma concentration at time zero of 60.63 +/- 28.26 microg/ml, with average plasma drug levels of 2.82 +/- 0.71 microg/ml at the end of the 192-hr sampling period . Plasma volume of distribution for i.v . oxytetracycline was 0.20 +/- 0.09 L/kg, with a harmonic mean elimination half-life of 15.15 hr and mean total body clearance rate of 0.007 +/- 0.002 L/hr/kg . Intramuscular administration of oxytetracycline achieved a mean peak plasma concentration of 6.85 +/- 1.96 microg/ml at 1 hr after administration, with average plasma drug levels of 4.96 +/- 1.97 microg/ml at the end of the 192-hr sampling period . The harmonic mean terminal elimination half-life for i.m . oxytetracycline was 131.23 hr . Based on the results of this study, long-acting preparations of oxytetracycline administered parenterally to American alligators at 10 mg/kg q 5 days is expected to maintain plasma concentrations above the minimum inhibitory concentration of 4.0 microg/ml for susceptible organisms.

J Zoo Wildl Med, 2004 Sep, 35(3), 333 - 40
Pharmacokinetics of enrofloxacin after single-dose oral and intravenous administration in the American alligator (Alligator mississippiensis); Helmick KE et al.; The pharmacokinetics of enrofloxacin administered orally and i.v . to American alligators (Alligator mississippiensis) at 5 mg/kg was determined . Plasma levels of enrofloxacin and its metabolite ciprofloxacin were measured using high-performance liquid chromatography and the resulting concentration versus time curve analyzed using compartmental modeling techniques for the i.v . data and noncompartmental modeling techniques for the oral data . A two-compartment model best represented the i.v . data . Intravenous administration of enrofloxacin resulted in an extrapolated mean plasma concentration of 4.19 +/- 4.23 microg/ml at time zero, with average plasma drug levels remaining above 1.0 microg/ml for an average of 36 hr . Plasma volume of distribution for i.v . enrofloxacin was 1.88 +/- 0.96 L/kg, with a harmonic mean elimination half-life of 21.05 hr and mean total body clearance rate of 0.047 +/- 0.021 L/hr/kg . Plasma levels of p.o . enrofloxacin remained below 1.0 microg/ml in all test animals, and average concentrations ranged from 0.08 to 0.50 microg/ml throughout the sampling period . Oral administration of enrofloxacin achieved a mean maximum plasma concentration of 0.50 +/- 0.27 microg/ml at 55 +/- 29 hr after administration, with a harmonic mean terminal elimination half-life of 77.73 hr . Minimal levels of ciprofloxacin were detected after both oral and i.v . enrofloxacin administration, with concentrations below minimum inhibitory concentrations for most susceptible organisms . On the basis of the results of this study, enrofloxacin administered to American alligators at 5 mg/kg i.v . q 36 hr is expected to maintain plasma concentrations that approximate the minimum inhibitory concentration for susceptible organisms (0.5 microg/ml) . Enrofloxacin administered to American alligators at 5 mg/kg p.o . is not expected to achieve minimum inhibitory values for susceptible organisms.

Acta Pharmacol Sin, 2004 Nov, 25(11), 1396 - 401
Tissue distribution of bitespiramycin and spiramycin in rats; Shi XG et al.; AIM: To investigate the tissue distribution of bitespiramycin (BSPM) and spiramycin (SPM) in rats . METHODS: Liquid chromatographic-mass spectrometric assay was applied for the determination of three major components (isovalerylspiramycins, ISV-SPMs) of BSPM and their major active metabolites (SPMs) in rat tissues and plasma after an oral dose of bitespiramycin, as well as SPMs . RESULTS: High levels of drug concentrations were observed in most tissues, especially in the liver, stomach, intestine, spleen, lung, womb, and pancreas . BSPM persisted long time in many rat tissues such that the drug concentration in spleen was 69.4 nmol/g at 60 h post-dose and it was still above the minimum inhibitory concentration of many susceptible pathogens . At 2.5 h post-dose, the total concentrations of ISV-SPMs and SPMs achieved in tissues were from 6 to 215 times higher than the corresponding concentrations in plasma . At 2.5 h post-dose, the mean C(t)/C(p) of BSPM appeared to be 2- or 3-fold those of SPM in most tissues . The tissue to plasma concentration ratios following oral dose of BSPM were higher than those of SPM in most tissues . The drug was not detected in brain and testis after a single dose of BSPM and SPM . CONCLUSION: Both BSPM and SPM penetrate into rat tissues well and BSPM has higher tissue affinity than SPM.

Arch Pediatr Adolesc Med, 2004 Nov, 158(11), 1070 - 6
Asthma and lung function 20 years after wheezing in infancy: results from a prospective follow-up study; Piippo-Savolainen E et al.; OBJECTIVE: To determine the outcome until adulthood after wheezing in infancy, compared with pneumonia in infancy and with controls . DESIGN: An 18- to-20-year prospective cohort study . SETTING: Pediatric department at a university hospital, providing primary hospital care for a defined population.Patients Fifty-four children hospitalized for bronchiolitis and 34 for pneumonia at younger than 2 years, and 45 controls with no early-life wheezing or hospitalization, were studied at median age 19 years . MAIN OUTCOME MEASURES: A questionnaire on asthma symptoms and medication, physical examination, flow volume spirometry (FVS), methacholine inhalation challenge (MIC), home peak expiratory flow (PEF) monitoring, and skin prick testing (SPT) to common inhalant allergens . The 2 asthma definitions were physician-diagnosed asthma and previously diagnosed asthma with recent asthmatic symptoms (physician-diagnosed asthma included) . RESULTS: By the 2 definitions, asthma was present in 30% (odds ratio {OR}, 3.37; 95% confidence interval {CI}, 1.12-10.10) and in 41% (OR 1.38; 95% CI, 0.37-5.21) in the bronchiolitis group, in 15% (OR, 5.50; 95% CI, 1.87-16.14) and in 24% (OR, 2.07; 95% CI, 0.59-7.22) in the pneumonia group, and in 11% in the control group . After bronchiolitis, the FVS values were forced vital capacity (FVC), 108% (SD, 13%) of predicted; forced expiratory volume in 1 second, 98% (SD, 12%); forced expiratory volume in 1 second divided by FVC, 91% (SD, 7.6%); midexpiratory flow at 50% of the FVC, 74% (SD, 19%); and midexpiratory flow at 25% of the FVC, 74% (SD, 22%) . Bronchial reactivity by MIC was present in 25 (48%) of 52 subjects in the bronchiolitis group, in 13 (42%) of 31 in the pneumonia group, and in 14 (32%) of 44 in the control group . The prevalence of atopy (positive SPTs) was 48% to 63% in the 3 groups . In a logistic regression adjusted for atopy and smoking, infantile bronchiolitis was an independent risk factor for asthma by both definitions . CONCLUSION: The increased risk for asthma persists until adulthood after bronchiolitis in infancy.

J Vet Intern Med, 2004 Sep-Oct, 18(5), 728 - 33
Pharmacokinetics of once-daily amikacin in healthy foals and therapeutic drug monitoring in hospitalized equine neonates; Bucki EP et al.; The objectives of this study were to investigate the pharmacokinetics of once-daily amikacin in healthy neonates, to determine amikacin concentrations in hospitalized foals, and to determine the minimum inhibitory concentrations (MICs) of amikacin against gram-negative isolates from blood cultures in septic foals . Median half-life, clearance, and volume of distribution of amikacin in healthy 2- to 3-day-old foals after administration of an intravenous bolus of amikacin (25 mg/kg) were 5.07 hours (4.86-5.45 hours), 1.82 mL/min/kg (1.35-1.97 mL/min/kg), and 0.785 L/kg (0.638-0.862 L/kg), respectively . Statistically significant (P <.05) decreases in area under the curve (14% decrease), mean residence time (19% decrease), and C24h plasma amikacin concentrations (29% decrease) occurred between days 2-3 and 10-11 . Plasma amikacin concentrations in healthy foals at 0.5 hours (C0.5h) were significantly higher (P = .02) than those of hospitalized foals . Sepsis, prematurity, and hypoxemia did not alter amikacin concentrations . The MIC at which 90% of all gram-negative isolates from equine neonatal blood cultures were inhibited by amikacin was 4 microg/mL, suggesting that amikacin C0.5h of 40 microg/mL should be targeted to achieve a maximum serum concentration to MIC ratio of 10:1 . The proportion of foals with C0.5h 40 microg/mL was significantly higher (P < .0001) in hospitalized foals receiving a dose of amikacin at 25 mg/kg (22/24 or 92%) than in foals receiving a dose at 21 mg/kg (9/25 or 36%), whereas no difference was found in the proportion of foals with C24h concentrations > or = 3 microg/mL between the 2 groups . An initial dose at 25 mg/kg is recommended for once-daily amikacin in equine neonates.

Med Clin (Barc), 2004 Oct 16, 123(13), 481 - 5
{Helicobacter pylori resistance to metronidazole and clarythromicin: descriptive analysis 1997-2000}; Gomollon F et al.; BACKGROUND AND OBJECTIVE: We aimed to assess the resistance of H . pylori to clarithromycin and metronidazole, in patients with and without previous eradication treatment, in a geographic area from the north of Spain . We also analyzed the evolution of resistance rates and its relationships with annual antibiotic consumption . PATIENTS AND METHOD: Retrospective study including all patients with H . pylori infection and positive culture from January 1997 to December 2000 . Minimal inhibitory concentrations (MIC) determined by the E test were used to report the clarithromycin (MIC > 2 mg/l) and metronidazole (MIC > 32 mg/l) resistance . RESULTS: A total of 537 clinical H . pylori isolates from patients without (n = 389) and with previous eradication treatment (n = 148) were studied . H . pylori resistance to clarithromycin and metronidazole was found in 8.7% (95% CI, 6.1-12) and 13.8% (95% CI, 10.4-17.3) patients without previous eradication treatment and in 39.2% (95% CI, 31.3-47.1) and 37.8% (95% CI, 30-45.7) patients with previous eradication treatment (p < 0.001), respectively . Clarithromycin resistance remained stable (1997: 9.7%; 1998: 5.7%; 1999: 11.8%; 2000: 6.2%) whereas metronidazole resistance decreased over the 4 years study period (1997: 38.7%; 1998: 15.1%; 1999: 9%; 2000: 6.9%) . We did not observe any clear relationship between resistance's evolution and antibiotic annual consumption . CONCLUSIONS: In our geographic area, primary resistance rates for clarithromycin remained stable whereas resistance for metronidazole decreased over the 4 years period.

Chang Gung Med J, 2004 Jul, 27(7), 475 - 88
Acute otitis media in children: current epidemiology, microbiology, clinical manifestations, and treatment; Leibovitz E et al.; An accurate differential diagnosis of AOM is essential for ensuring appropriate treatment, since overdiagnosis of disease is common and antibiotics are not indicated for otitis media with effusion . Although antibiotic therapy is required in only 20-30% of all AOM cases (high rate of spontaneous recovery), most of the patients are treated since this small proportion cannot be quickly and easily identified . The main determinant of the efficacy of antibiotics in AOM is the time that drug concentration at the site of infection exceeds the minimal inhibitory concentration for the pathogen . The major problems encountered in the antibiotic therapy of AOM are the tremendous increase in the resistance to antibiotics of its main pathogens and the lack of tight criteria in the selection of the appropriate antibiotic drugs for the treatment of this disease . The recently published Center for Disease Control and Prevention (CDC) guidelines for the treatment of AOM represent a major step forward in the rational approach to the management of this disease by establishing a clear hierarchy among the various therapeutic agents used in the treatment of simple and complicated AOM . A seven-valent pneumococcal conjugate vaccine recently licensed in the United States for universal immunization of infants <2 years has demonstrated efficacy for prevention of serotype-specific pneumococcal AOM.

J Ethnopharmacol, 2004 Dec, 95(2-3), 253 - 8
In vitro biological activity and essential oil composition of four indigenous South African Helichrysum species; Lourens AC et al.; Helichrysum species are used widely to treat various medical conditions . In this study, the anti-microbial, anti-oxidant (DPPH assay) and anti-inflammatory activity (5-lipoxygenase assay) of Helichrysum dasyanthum, Helichrysum felinum, Helichrysum excisum and Helichrysum petiolare were investigated . The essential oil compositions of these species were determined . The acetone and methanol extracts as well as the essential oils exhibited activity against Gram-positive bacteria, while both the methanol and acetone extracts of all four species were active in the anti-oxidant assay . The essential oils, on the other hand, displayed activity in the 5-lipoxygenase assay, which was used as an indication of anti-inflammatory activity . Two extracts exhibited promising activity in the anti-microbial assay, the acetone extract of Helichrysum dasyanthum with a MIC value of 15.63 microg/ml and the methanol extract of Helichrysum excisum with a MIC value of 62.5 microg/ml . The acetone extract of Helichrysum dasyanthum was the most active free radical scavenger in the DPPH assay (IC(50) of 9.53 microg/ml) while values for the anti-inflammatory activity of the essential oils ranged between 25 and 32 microg/ml . The essential oil compositions of three species (Helichrysum dasyanthum, Helichrysum excisum and Helichrysum petiolare) were dominated by the presence of monoterpenes such as alpha-pinene, 1,8-cineole and p-cymene . In the oil of Helichrysum felinum, monoterpenes were largely absent . Its profile consisted of a variety of sesquiterpenes in low concentrations with beta-caryophyllene dominating.

Antimicrob Agents Chemother, 2004 Nov, 48(11), 4246 - 9
Concentrations of gemifloxacin at the target site in healthy volunteers after a single oral dose; Islinger F et al.; Free gemifloxacin concentrations in the interstitial space fluid of skeletal muscle and subcutaneous adipose tissue were measured by means of in vivo microdialysis to characterize the ability of gemifloxacin to penetrate human soft tissues . Twelve healthy volunteers received a single oral dose of 320 mg of gemifloxacin . The mean areas under the concentration-time curves from 0 to 10 h (AUC(0-10)) were significantly higher for soft tissue than for unbound gemifloxacin in plasma (P < 0.05) . The ratios of the mean AUC(0-10) for tissue to the AUC(0-10) for free gemifloxacin in plasma were 1.7 +/- 0.7 (mean +/- standard deviation) for skeletal muscle and 2.4 +/- 1.0 for adipose tissue . The AUC(0-24) ratios for free gemifloxacin in tissues to the MIC at which 90% of frequently isolated bacteria are inhibited were close to or higher than 100 h . Therefore, based on pharmacokinetic and pharmacodynamic calculations, we conclude that gemifloxacin might be a useful therapeutic option for the treatment of soft tissue infections.

Antimicrob Agents Chemother, 2004 Nov, 48(11), 4163 - 70
Multidrug resistance of a porin deletion mutant of Mycobacterium smegmatis; Stephan J et al.; Mycobacteria contain an outer membrane of unusually low permeability which contributes to their intrinsic resistance to many agents . It is assumed that small and hydrophilic antibiotics cross the outer membrane via porins, whereas hydrophobic antibiotics may diffuse through the membrane directly . A mutant of Mycobacterium smegmatis lacking the major porin MspA was used to examine the role of the porin pathway in antibiotic sensitivity . Deletion of the mspA gene caused high-level resistance of M . smegmatis to 256 microg of ampicillin/ml by increasing the MIC 16-fold . The permeation of cephaloridine in the mspA mutant was reduced ninefold, and the resistance increased eightfold . This established a clear relationship between the activity and the outer membrane permeation of cephaloridine . Surprisingly, the MICs of the large and/or hydrophobic antibiotics vancomycin, erythromycin, and rifampin for the mspA mutant were increased 2- to 10-fold . This is in contrast to those for Escherichia coli, whose sensitivity to these agents was not affected by deletion of porin genes . Uptake of the very hydrophobic steroid chenodeoxycholate by the mspA mutant was retarded threefold, which supports the hypothesis that loss of MspA indirectly reduces the permeability by the lipid pathway . The multidrug resistance of the mspA mutant highlights the prominent role of outer membrane permeability for the sensitivity of M . smegmatis to antibiotics . An understanding of the pathways across the outer membrane is essential to the successful design of chemotherapeutic agents with activities against mycobacteria.

Vet J, 2004 Nov, 168(3), 312 - 6
Disposition of norfloxacin in broiler chickens and turkeys after different methods of oral administration; Sarkozy G et al.; Norfloxacin was administered orally to chickens and turkeys at 15 mg/kg body weight by pulse dosing at 24 h intervals and by continuous dosing at 100 mg/L in drinking water for five days . Blood samples were taken serially . Plasma norfloxacin concentrations were determined by high-performance liquid chromatography . The plasma norfloxacin concentrations increased slowly during continuous dosing and reached the MIC(90) (250 ng/mL) for Gram-negative pathogens by 12 h in chickens and 18 h in turkeys . The steady-state plasma concentration was attained in 36 h and remained at approximately 776.67+/-33.23 ng/mL in chickens and 682.50+/-28.55 ng/mL in turkeys . After pulse dosing, the plasma norfloxacin concentrations increased rapidly and exceeded the MIC(90) at 2 h in both species and remained above MIC(90) for 8 h in chickens and 6 h in turkeys . Pulse dosing provided half the steady-state concentration that was achieved by continuous dosing, 365.32+/-39.31 ng/mL in chickens and 306.03+/-32.26 ng/mL in turkeys, during the dosing interval of 24 h . Data for daily pulse dosing suggested that every administration corresponded to a single, daily repeated bolus administration although pulse dosing produced higher plasma concentrations more readily . Continuous and pulse dosing are both rational for the administration of norfloxacin to flocks of chickens and turkeys . We recommend that treatment be commenced with a pulse oral dose administered over a 4 h period and maintained by continuous oral medication for three to five consecutive days.

J Org Chem, 2004 Oct 29, 69(22), 7428 - 35
Eremophilane sesquiterpenes from capsidiol; Zhao Y et al.; A series of eremophilane sesquiterpene alcohols and hydrocarbons was prepared from the phytoalexin capsidiol (1) for mechanistic studies with epiaristolochene synthase and epiaristolochene dihydroxylase . Among them, 3-deoxycapsidiol (10) was obtained through selective derivatization and reductive cleavage of the equatorial 3 alpha hydroxyl group . Two novel isomers of aristolochene and eremophilene were accessed from the 1- and 3-deoxycapsidiol isomers . 4-Epieremophilene (17) was obtained by conjugate reduction of epiaristolochen-1-one tosylhydrazone with catecholborane followed by sulfinate elimination and diimide rearrangement . Epimerization of epiaristolochen-3-one (27a) at the C4 methyl followed by reductions led to the previously unknown aristolochene isomer, eremophila-9(10),11(12)-diene (30) . Optical rotations and characteristic (1)H NMR data for the related eremophilenols and dienes are collected in Tables 1 and 2 . Finally, bioassays were used to assess the antifungal potencies of capsidiol and its synthetic derivatives . The minimum inhibitory concentration for capsidiol (3-10 ng) was at least 1 order of magnitude lower than that of any of the derivatives and considerably lower than those previously reported for ketoconazole, nystatin, and propiconazole.

Biomedica, 2004 Jun, 24 Supp 1, 85 - 91
In vitro susceptibility testing of Mycobacterium tuberculosis complex strains isolated from seals to antituberculosis drugs; Bernardelli A et al.; Mycobacteria strains belonging to the Mycobacterium tuberculosis complex were isolated from seals found in the South Atlantic . The animals were received in Mundo Marino installations and treated for Mycobacterium tuberculosis complex by conventional therapy of intensive care and enriched food supply; however, in all cases treatment failed . Necropsies of all animals revealed extensive lesions compatible with tuberculosis involving lungs, liver, spleen and lymphatic nodes . Classical biochemical methods as well as molecular techniques using the IS6110 probes were performed for mycobacterial identification . Furthermore, the LCx M . tuberculosis assay (Abbott Laboratories) identified all strains as Mycobacterium tuberculosis complex members . The in vitro susceptibility pattern was examined in mycobacterial strains isolated from seven seals and in 3 reference strains--BCG, H37Rv (M . tuberculosis) and AN5 (Mycobacterium bovis)--to 4 medications--isoniazid, rifampin, streptomycin and ethambutol . Minimal inhibitory drug concentrations were determined by the Mycobacterial Growth Indicator Tube (BD Argentina) method and a microdilution and colorimetric assay using 3-(4-5 dimethyltiazol-2)-2,5 diphenyltetrazolium bromide . All the isolates and the reference strains BCG and AN5 were inhibited by MIC values similar to those of H37Rv with good agreement obtained by both techniques . These findings suggest that a therapeutic regimen aimed to seals diagnosed with tuberculosis play an important role in the prevention of tuberculosis transmission from infected animals to humans that are in routine contact with them.

Langmuir, 2004 Oct 26, 20(22), 9551 - 9
Thermodynamics of micellization of benzyl(2-acylaminoethyl)dimethylammonium chloride surfactants in aqueous solutions: a conductivity and titration calorimetry study; Shimizu S et al.; The enthalpies of micellization of the surfactant series benzyl(2-acylaminoethyl)dimethylammonium chlorides, RABzMe(2)Cl, have been determined by calorimetry and conductivity measurements in the temperature range 15-75 degrees C . Here R stands for an acyl group containing 10-16 carbon atoms and A, Bz, and Me stand for NH(CH(2))(2)N(+), benzyl, and methyl groups, respectively . The enthalpy of micellization, DeltaH(mic) degrees , and the critical micelle concentration, cmc, were calculated directly from calorimetric data . The free energy of micellization, DeltaG(mic) degrees , was obtained from the cmc and the conductance-based degree of counterion dissociation . There is an excellent agreement between DeltaG(mic) degrees calculated from the data of both techniques, but the DeltaH(mic) degrees , the entropy of micellization, values differ . The dependence of the thermodynamic parameters of micellization on the chain length of the hydrophobic group and on the temperature has been analyzed by considering the delicate balance between the factors that contribute to micelle formation, including transfer of the surfactant hydrocarbon chain from the aqueous environment to the micelle, with concomitant release of the solvating water molecules, and the effect of temperature on the structure of water . DeltaG(mic) degrees is more negative, that is, more favorable for RABzMe(2)Cl than for the structurally related alkylbenzyldimethylammonium chlorides . This is attributed to direct and water-mediated H bonding between the amide groups of molecules of the former series.

Pest Manag Sci, 2004 Oct, 60(10), 1007 - 12
Synthesis and fungicidal activity of ethaboxam against Oomycetes; Kim DS et al.; This study describes the chemical synthesis and intrinsic fungicidal activity of ethaboxam {(RS)-N-(alpha-cyano-2-thenyl)-4-ethyl-2-(ethylamino)-1,3-thiazole-5-carboxamide}, a new Oomycetes fungicide . In in vitro tests, ethaboxam showed inhibitory activity against isolates of Phytophthora and some Pythium spp, with MIC values ranging from 0.1 to 0.5 mg litre(-1) for nine isolates of Phytophthora infestans (Montagne) de Bary and from 1.0 to 5.0 mg litre(-1) for eight isolates of Phytophthora capsici Leonian . In tests to determine time and concentration for complete inactivation of each pathogen (five isolates of P infestans and five isolates of P capsici), ethaboxam inactivated all isolates of P infestans within 48h at 10 mg litre(-1) and those of P capsici within 96 h at 10 mg litre(-1) . Ethaboxam effectively suppressed development of tomato late blight caused by P infestans and pepper Phytophthora blight caused by P capsici in the studies conducted to determine its preventive, curative, persistent and systemic activity . These results show that ethaboxam has desirable fungicidal characteristics as an Oomycetes fungicide.

Phytother Res, 2004 Sep, 18(9), 713 - 7
The chemical composition of some Lauraceae essential oils and their antifungal activities; Simic A et al.; The antifungal activity of Aniba rosaeodora, Laurus nobilis, Sassafras albidum and Cinnamomum zeylanicum essential oils were investigated against 17 micromycetes . Among the tested fungal species were food poisoning, spoilage fungi, plant and animal pathogens . In order to determine fungistatic and fungicidal concentrations (MIC and MFC) macrodilution and microdilution tests were used . Linalool was the main component in the essential oil of A . rosaeodora, while 1.8-cineole was dominant in L . nobilis . In sassafras essential oil safrole was the major component and in the oil of C . zeylanicum the main component was trans-cinnamaldehyde . The essential oil of cinnamon showed the strongest antifungal activity . Copyright (c) 2004 John Wiley & Sons, Ltd.

Cancer Biol Ther . 2004 Dec 14;3(12) {Epub ahead of print}
Differentially Expressed Genes in Pancreatic Ductal Adenocarcinomas Identified Through Serial Analysis of Gene Expression; Hustinx SR et al.; Serial analysis of gene expression (SAGE) is a powerful tool for the discovery of novel tumor markers . The publicly available online SAGE libraries of normal and neoplastic tissues have recently been expanded; in addition, a more complete annotation of the human genome and better biocomputational techniques have substantially improved the assignment of differentially expressed SAGE "tags" to human genes . These improvements have provided us with an opportunity to re-evaluate global gene expression in pancreatic cancer using existing SAGE libraries . SAGE libraries generated from six pancreatic cancers were compared to SAGE libraries generated from 11 non-neoplastic tissues . Compared to normal tissue libraries, we identified 453 SAGE tags as differentially expressed in pancreatic cancer, including 395 that mapped to known genes and 58 "uncharacterized" tags . Of the 395 SAGE tags assigned to known genes, 223 were overexpressed in pancreatic cancer, and 172 were underexpressed . In order to map the 58 uncharacterized differentially expressed SAGE tags to genes, we used a newly developed resource called TAGmapper , to identify 16 additional differentially expressed genes . The differential expression of seven genes, involved in multiple cellular processes such as signal transduction (MIC-1), differentiation (DMBT1 and Neugrin), immune response (CD74), inflammation (CXCL2), cell cycle (CEB1) and enzymatic activity (Kallikrein 6), was confirmed by either immunohistochemical labeling of tissue microarrays (Kallikrein 6, CD74 and DMBT1) or by RT-PCR (CEB1, Neugrin, MIC1 and CXCL2) . Of note, Neugrin was one of the genes whose previously uncharacterized SAGE tag was correctly assigned using TAGmapper, validating the utility of this program . Novel differentially expressed genes in a cancer type can be identified by revisiting updated and expanded SAGE databases . TAGmapper should prove to be a powerful tool for the discovery of novel tumor markers through assignment of uncharacterized SAGE tags.

Arch Pharm (Weinheim), 2004 Oct, 337(10), 549 - 55
Synthesis and antimycobacterial activity of pyridylmethylsulfanyl and naphthylmethylsulfanyl derivatives of benzazoles, 1, 2, 4-triazole, and pyridine-2-carbothioamide/-2-carbonitrile; Zahajska L et al.; A set of four types of benzazoles, 1, 2, 4-triazole, and pyridine-2-carbonitrile/-2-carbothioamide substituted with 1-naphthylmethylsulfanyl or pyridylmethylsulfanyl was prepared to modify the structure of benzylsulfanyl derivatives of the above-mentioned heterocycles . The compounds were evaluated for in vitro antimycobacterial activity against Mycobacterium tuberculosis, M . avium, and two strains of M . kansasii . The activities were expressed as the minimum inhibitory concentration (MIC) . The MIC values fall into a range of 2 to >1000 micromol/L . Introduction of a pyridyl moiety into the molecule mostly decreased the activity . A naphthyl moiety did not influence the activity in comparison with a phenyl . The most active substances were 4-(3-pyridylmethylsulfanyl)pyridine-2-carbothioamide (7b) (MIC = 2 - >62.5 micromol/L) and 4-(1-naphthylmethylsulfanyl)pyridine-2-carbothioamide (7d) (MIC = 2 - >32 micromol/L).

Clin Cancer Res, 2004 Oct 1, 10(19), 6418 - 31
Immediate gene expression changes after the first course of neoadjuvant chemotherapy in patients with primary breast cancer disease; Modlich O et al.; PURPOSE: Our goal was to identify genes undergoing expressional changes shortly after the beginning of neoadjuvant chemotherapy for primary breast cancer . EXPERIMENTAL DESIGN: The biopsies were taken from patients with primary breast cancer prior to any treatment and 24 hours after the beginning of the neoadjuvant chemotherapy . Expression analyses from matched pair samples representing 25 patients were carried out with Clontech filter arrays . A subcohort of those 25 paired samples were additionally analyzed with the Affymetrix GeneChip platform . All of the transcripts from both platforms were queried for expressional changes . RESULTS: Performing hierarchical cluster analysis, we clustered pre- and posttreatment samples from individual patients more closely to each other than the samples taken from different patients . This reflects the rather low number of transcripts responding directly to the drugs used . Although transcriptional drug response occurring during therapy differed between individual patients, two genes (p21(WAF1/CIP1) and MIC-1) were up-regulated in posttreatment samples . This could be validated by semiquantitative and real-time reverse transcription-PCR . Partial least- discriminant analysis based on approximately 25 genes independently identified by either Clontech or Affymetrix platforms could clearly discriminate pre- and posttreatment samples . However, correlation of certain gene expression levels as well as of differential patterns and clusters as determined by a different platform was not always satisfying . CONCLUSIONS: This study has demonstrated the potential of monitoring posttreatment changes in gene expression as a measure of the pharmacodynamics of drugs . As a clinical laboratory model, it can be useful to identify patients with sensitive and reactive tumors and to help for optimized choice for sequential therapy and obviously improve relapse- free and overall survival.

J Cataract Refract Surg, 2004 Oct, 30(10), 2177 - 82
Comparative penetration of moxifloxacin and gatifloxacin in rabbit aqueous humor after topical dosing; Levine JM et al.; PURPOSE: To evaluate the aqueous penetration of the fourth-generation fluoroquinolones moxifloxacin and gatifloxacin . SETTING: University of Arizona, Tucson, Arizona, USA . METHODS: Forty eyes of 20 New Zealand white rabbits were divided into 2 experimental groups . In Experiment I rabbits (20 eyes), a commercial preparation of topical gatifloxacin 0.3% was administered to 9 eyes and moxifloxacin 0.5% to 9 eyes; 2 eyes served as a control . Eyes were dosed according to a keratitis protocol; ie, every 15 minutes for 4 hours . The aqueous humor was sampled 10 minutes after the last dose . Experiment II rabbits (20 eyes) were dosed according to a cataract prophylaxis protocol; ie, 4 times a day for 10 days . The aqueous humor was sampled 1 hour after the last dose of antibiotic in 12 eyes and 24 hours after the last dose in 8 eyes . High-performance liquid chromatography was used to determine the fluoroquinolone concentration . RESULTS: In the keratitis dosing protocol, the mean concentration of moxifloxacin in the aqueous (n=9) was 11.057 microg/mL (range 7.66 to 18.87 microg/mL), which was significantly higher than the mean concentration of gatifloxacin (n=8) (7.570 microg/mL {range 4.75 to 10.86 microg/mL}) (P=.030) . In the cataract prophylaxis dosing protocol, the mean aqueous concentration of moxifloxacin (n=6) was 1.745 microg/mL (range 0.92 to 3.87 mg/mL) . The mean concentration of gatifloxacin (n=6) was 1.207 microg/mL (range 0.44 to 2.44 microg/mL) . The difference was not statistically significant (P=.359) . CONCLUSIONS: Higher mean levels (x1.46) of aqueous penetration were achieved with moxifloxacin than with gatifloxacin in the keratitis-dosing model . There was no statistically significant difference between the 2 drugs in the cataract prophylaxis dosing model . Both antibiotics had aqueous levels in excess of the minimum inhibitory concentration for most pathogenic organisms in both models.

J Clin Microbiol, 2004 Oct, 42(10), 4577 - 80
Clinical evaluation of the Sensititre YeastOne colorimetric antifungal plate for antifungal susceptibility testing of the new triazoles voriconazole, posaconazole, and ravuconazole; Pfaller MA et al.; A commercially prepared dried colorimetric microdilution panel (Sensititre YeastOne, TREK Diagnostic Systems, Cleveland, Ohio) was compared in three different laboratories with the National Committee for Clinical Laboratory Standards (NCCLS) reference microdilution method by testing two quality control strains and 300 clinical isolates of Candida spp . against fluconazole, voriconazole, posaconazole, and ravuconazole . Reference MIC endpoints were established after 48 h of incubation and YeastOne colorimetric endpoints were established after 24 h of incubation . YeastOne endpoints were determined to be the lowest concentration at which the color in the well changed from red (indicating growth) to purple (indicating growth inhibition) or blue (indicating no growth) . Excellent agreement (within two dilutions) between the reference and colorimetric MICs was observed . Overall agreement was 95.4% . Agreement ranged from 92.3% with posaconazole to 98.0% with fluconazole . The YeastOne colorimetric method appears to be comparable to the NCCLS reference method for testing the susceptibility of Candida spp to the new triazoles voriconazole, posaconazole, and ravuconazole.

J Antimicrob Chemother, 2004 Nov, 54(5), 940 - 3 Epub 2004 Oct 07.
Post-antifungal effect of amphotericin B and voriconazole against Aspergillus fumigatus analysed by an automated method based on fungal CO2 production: dependence on exposure time and drug concentration; Chryssanthou E et al.; The post-antifungal effect (PAFE) of amphotericin B and voriconazole, either alone or in combination, on Aspergillus fumigatus was studied using an automated system based on fungal CO(2) production . METHODS: Conidia of A . fumigatus were exposed to concentrations of 1-10 x MIC of amphotericin B and 1-40 x MIC of voriconazole for 1, 2 and 4 h . After a washing step, exposed and control conidia were inoculated into Pedi-BacT culture bottles . CO(2) production was automatically monitored until the bottles signalled positive . The difference in time span for positive signals in drug-exposed and control bottles was used to calculate PAFE . RESULTS: There was a linear relationship between inoculum size and time to positive signal (r(2)=0.99) . The precision of duplicate analyses was 1.5% . Longer exposure times increased the amphotericin B-induced PAFE (P<0.001), whereas concentrations above the MIC did not . Voriconazole after 4 h of exposure induced a short dose-independent PAFE . The combination with amphotericin B did not prolong the PAFE over that caused by amphotericin B alone . CONCLUSIONS: This automated method can be used for determination of PAFE . In contrast to Candida spp., in which amphotericin B-induced PAFE is mainly related to the area under the curve, the effect on A . fumigatus was more dependent on the exposure time . This implies that pharmacodynamic data obtained from Candida experiments cannot be directly extrapolated to Aspergillus.

Bioorg Med Chem, 2004 Nov 1, 12(21), 5651 - 9
Synthesis and anti-tubercular activity of a series of 2-sulfonamido/trifluoromethyl-6-substituted imidazo{2,1-b}-1,3,4-thiadiazole derivatives; Gadad AK et al.; A series of 2-sulfonamido/trifluoromethyl-6-(4'-substituted aryl/heteroaryl)imidazo{2,1-b}-1,3,4-thiadiazole derivatives (II) have been synthesized by reaction of 2-amino-5-sulfonamido/trifluoromethyl-1,3,4-thiadiazoles and an appropriate alpha-haloaryl/heteroaryl ketones . Further 5-bromo (III), 5-thiocyanato (IV), 5-gaunylhydrazone (V) derivatives were synthesized in order to study the effect of these substituents on biological activity . Structures of these compounds were established by IR, (1)H NMR, (13)C NMR, Mass and HRMS . The selected compounds were evaluated for their preliminary in vitro anti-tuberculosis activity against Mycobacterium tuberculosis H37Rv strain using radiometric BACTEC and broth dilution assay methods . The results show that compounds 5, 7, 8, 10 and 12 exhibited moderate to good anti-tubercular activity with percentage inhibition of 29, 43, 58, 31 and 41, respectively, at a MIC of >6.25 microg/mL . Compound 18 showed a MIC of 20 microg/mL.

Stud Health Technol Inform, 2002, 90, 277 - 81
The Cadmio XML healthcare record; Barbera F et al.; The management of clinical data is a complex task . Patient related information reported in patient folders is a set of heterogeneous and structured data accessed by different users having different goals (in local or geographical networks) . XML language provides a mechanism for describing, manipulating, and visualising structured data in web-based applications . XML ensures that the structured data is managed in a uniform and transparent manner independently from the applications and their providers guaranteeing some interoperability . Extracting data from the healthcare record and structuring them according to XML makes the data available through browsers . The MIC/MIE model (Medical Information Category/Medical Information Elements), which allows the definition and management of healthcare records and used in CADMIO, a HISA based project, is described in this paper, using XML for allowing the data to be visualised through web browsers.

Antibiot Khimioter, 2004, 49(4), 3 - 6
{Lovastatin effect on growth of Tolypocladium inflatum 106 and ergosterol production by the culture}; Involvement of growth differentiation factor-15/macrophage inhibitory cytokine-1 (GDF-15/MIC-1) in oxLDL-induced apoptosis of human macrophages in vitro and in arteriosclerotic lesions; Department of Anatomy and Cell Biology III, Interdisciplinary Center of Neurosciences (IZN), University of Heidelberg, Im Neuenheimer Feld 307, 69120 Heidelberg, GermanyGrowth differentiation factor-15/macrophage inhibitory cytokine-1 (GDF-15/MIC-1) is a new member of the transforming growth factor beta (TGF-beta) superfamily, which has most recently been found in activated macrophages (MPhi) . We have now investigated GDF-15/MIC-1 in human MPhi after exposure to oxidized low-density lipoproteins (oxLDL) related mediators in vitro and in arteriosclerotic carotid arteries . Using RT-PCR and Western blotting a pronounced induction of GDF-15/MIC-1 expression by oxLDL, C6-ceramide, tumor necrosis factor (TNFalpha) and hydrogen peroxide (H2O2) was found in cultured human MPhi . In 11 human arteriosclerotic carotid arteries, immunohistochemical analyses supported by computer-assisted morphometry and regression analyses demonstrated a significant colocalization of GDF-15/MIC-1 immunoreactivity (IR) with oxLDL IR and manganese superoxide dismutase (MnSOD) IR in CD68 immunoreactive (ir) MPhi, which were also expressing AIF-IR (apoptosis-inducing factor), caspase-3-IR (CPP32), PARP-IR, c-Jun/AP-1-IR and p53-IR . Our data suggest that GDF-15/MIC-1 is inducible in human MPhi by oxLDL and its mediators in vitro and is supposed to contribute to oxidative stress dependent consequences in arteriosclerotic plaques, e.g . modulating apoptosis and inflammatory processes in activated MPhi.

Rev Iberoam Micol, 2003 Jun, 20(2), 64 - 7
{Successful treatment with voriconazol of a Pseudallescheria boydii fungus ball in a HIV positive patient and previous tuberculosis}; Garci J et al.; We herein describe a patient with a Pseudallescheria boydii fungus ball in a tuberculous lung cavity, which was successfully treated four years earlier . The patient was HIV positive classified as C3 with a previous history of i.v . heroin abuse . The clinical presumptive diagnosis was radiologically established combined with histological examination . Culture of tissue confirmed and proved the fungal etiology . In vitro MIC values for voriconazole (0.5 mg/ml) guided antifungal prophylactic treatment before surgical eradication of the fungus ball since the patient was immunosuppressed . We discuss the clinical spectrum of P . boydii infections and currently medical approach.

J Med Chem, 2004 Oct 7, 47(21), 5276 - 83
Synthesis and evaluation of nitrofuranylamides as novel antituberculosis agents; Tangallapally RP et al.; In an effort to develop new and more potent therapies to treat tuberculosis, a library of compounds was screened for M . tuberculosis UDP-Gal mutase inhibition . Nitrofuranylamide 1 was identified as a hit in this screen, possessing good antituberculosis activity . This paper describes the synthesis and evaluation of an expanded set of nitrofuranylamides . We have discovered a number of nitrofuranylamides with submicromolar M . tuberculosis MIC values and acceptable therapeutic indexes . The MIC activity did not correlate with UDP-Gal mutase inhibition, suggesting an alternative primary cellular target was responsible for the antituberculosis activity . The compounds were only active against mycobacteria of the tuberculosis complex . On the basis of these results, four compounds were selected for in vivo testing in a mouse model of tuberculosis infection, and of these compounds one showed significant antituberculosis activity.

BMC Bioinformatics . 2004 Sep 28;5(1):137.
Few amino acid positions in rpoB are associated with most of the rifampin resistance in Mycobacterium tuberculosis; Cummings MP et al.; BACKGROUND: Mutations in rpoB, the gene encoding the beta subunit of DNA-dependent RNA polymerase, are associated with rifampin resistance in Mycobacterium tuberculosis . Several studies have been conducted where minimum inhibitory concentration (MIC, which is defined as the minimum concentration of the antibiotic in a given culture medium below which bacterial growth is not inhibited) of rifampin has been measured and partial DNA sequences have been determined for rpoB in different isolates of M . tuberculosis . However, no model has been constructed to predict rifampin resistance based on sequence information alone . Such a model might provide the basis for quantifying rifampin resistance status based exclusively on DNA sequence data and thus eliminate the requirements for time consuming culturing and antibiotic testing of clinical isolates . RESULTS: Sequence data for amino acid positions 511-533 of rpoB and associated MIC of rifampin for different isolates of M . tuberculosis were taken from studies examining rifampin resistance in clinical samples from New York City and throughout Japan . We used tree-based statistical methods and random forests to generate models of the relationships between rpoB amino acid sequence and rifampin resistance . The proportion of variance explained by a relatively simple tree-based cross-validated regression model involving two amino acid positions (526 and 531) is 0.679 . The first partition in the data, based on position 531, results in groups that differ one hundredfold in mean MIC (1.596 micrograms/ml and 159.676 micrograms/ml) . The subsequent partition based on position 526, the most variable in this region, results in a > 354-fold difference in MIC . When considered as a classification problem (susceptible or resistant), a cross-validated tree-based model correctly classified most (0.884) of the observations and was very similar to the regression model . Random forest analysis of the MIC data as a continuous variable, a regression problem, produced a model that explained 0.861 of the variance . The random forest analysis of the MIC data as discrete classes produced a model that correctly classified 0.942 of the observations with sensitivity of 0.958 and specificity of 0.885 . CONCLUSIONS: Highly accurate regression and classification models of rifampin resistance can be made based on this short sequence region . Models may be better with improved (and consistent) measurements of MIC and more sequence data.

J Health Soc Policy, 2004, 18(4), 1 - 11
Maternity and infant care, race and birth outcomes; Holian J et al.; We examine the effect of a large, comprehensive maternity and infant care (MIC) program on birthweight and infant mortality in an economically depressed urban population . The study is based on linked birth, infant death and program files for 1985-87 Cleveland and East Cleveland, Ohio, birth cohorts (N = 31,415) . Taking into account differences in risk factors, Black MIC infants experienced lower neonatal and endogenous mortality, but White MIC infants had higher postneonatal and exogenous mortality than their same race, non-MIC counterparts . Birthweight distributions were also more favorable for Black than White clients . We discuss the policy implications of our findings.

J Pharm Sci, 2004 Nov, 93(11), 2851 - 62
Effects of different N-trimethyl chitosans on in vitro/in vivo ofloxacin transcorneal permeation; Di Colo G et al.; N-trimethyl chitosan (TMC) polymers differing in quaternization degree (QD) and molecular weight (MW) were prepared from two chitosans 90% deacetylated, one of higher MW (1460 kDa) (TMCH), the other of lower MW (580 kDa) (TMCL), by one (TMCH1, QD = 4%; TMCL1, QD = 3%), two (TMCH2, QD = 35%; TMCL2, QD = 46%), or three (TMCH3, QD = 90%; TMCL3, QD = 78%) reductive methylation steps . The derivatives were tested and compared for their ability to enhance the permeability of ofloxacin across rabbit corneal epithelium, reconstituted in vitro . TMC polymers of intermediate QD (TMCH2 and TMCL2), at the concentration of 0.001% w/v, produced significant permeability enhancements, independent of polymer MW . The enhancing effect did not increase when QD was increased (TMCH3 and TMCL3), while it was not significant with low QD values (TMCH1 and TMCL1) . Such an effect was specific of chitosan derivatives, because fully quaternized DEAE-dextran (MeDD) was ineffective . The transcorneal permeability-enhancing property of TMCH2 and TMCL2, and the inefficacy of MeDD were confirmed by in vivo tests on rabbit eyes . However, unlike the in vitro experiments, the in vivo ones showed a stronger effect of the TMC having higher MW . TMCH2 produced antibiotic levels in the aqueous humor higher than the MIC(90%) for the more resistant ocular pathogens . These results point to this derivative as a potential ofloxacin absorption enhancer for the topical treatment of endophthalmitis.

Antimicrob Agents Chemother, 2004 Oct, 48(10), 4027 - 32
Antipneumococcal activity of LBM415, a new peptide deformylase inhibitor, compared with those of other agents; Ednie LM et al.; The MICs of LBM415, a new peptide diformylase inhibitor, were evaluated and ranged from 0.03 to 4.0 microg/ml for 300 pneumococci, irrespective of their beta-lactam, macrolide, and quinolone susceptibilities . By comparison, vancomycin, teicoplanin, linezolid, and quinupristin-dalfopristin were also active, with MICs </=2.0 microg/ml . Gatifloxacin and moxifloxacin were the most active quinolones tested, while the MICs of the beta-lactams rose with those of penicillin G . LBM415 at two times the MIC was bactericidal (99.9% killing) against six strains after 24 h.

Antimicrob Agents Chemother, 2004 Oct, 48(10), 4009 - 11
Correlation between in vitro susceptibility of Scedosporium apiospermum to voriconazole and in vivo outcome of scedosporiosis in guinea pigs; Capilla J et al.; We have evaluated the efficacy of voriconazole (VRC) in a systemic infection by Scedosporium apiospermum in immunodepressed guinea pigs . Animals were infected with two strains; one required a VRC MIC of 0.5 to 1 microg/ml, common for this fungus, and the other required a high MIC (8 microg/ml), unusual in this species . VRC prolonged survival and reduced fungal load in kidney and brain tissues of the animals infected with the first strain but was unable to prolong survival or to reduce fungal load in brain tissue for the latter strain.

Antimicrob Agents Chemother, 2004 Oct, 48(10), 3871 - 6
Genomic approach to identification of mutations affecting caspofungin susceptibility in Saccharomyces cerevisiae; Markovich S et al.; The antifungal agent caspofungin (CAS) specifically interferes with glucan synthesis and cell wall formation . To further study the cellular processes affected by CAS, we analyzed a Saccharomyces cerevisiae mutant collection (4,787 individual knockout mutations) to identify new genes affecting susceptibility to the drug . This collection was screened for increased CAS sensitivity (CAS-IS) or increased CAS resistance (CAS-IR) . MICs were determined by the broth microdilution method . Disruption of 20 genes led to CAS-IS (four- to eightfold reductions in the MIC) . Eleven of the 20 genes are involved in cell wall and membrane function, notably in the protein kinase C (PKC) integrity pathway (MID2, FKS1, SMI1, and BCK1), chitin and mannan biosynthesis (CHS3, CHS4, CHS7, and MNN10), and ergosterol biosynthesis (ERG5 and ERG6) . Four of the 20 genes (TPO1, VPS65, VPS25, and CHC1) are involved in vacuole and transport functions, 3 of the 20 genes (CCR4, POP2, and NPL3) are involved in the control of transcription, and 2 of the 20 genes are of unknown function . Disruption of nine additional genes led to CAS-IR (a fourfold increase of MIC) . Five of these nine genes (SLG1, ERG3, VRP1, CSG2, and CKA2) are involved in cell wall function and signal transduction, and two of the nine genes (VPS67 and SAC2) are involved in vacuole function . To assess the specificity of susceptibility to CAS, the MICs of amphotericin B, fluconazole, flucytosine, and calcofluor for the strains were tested . Seven of 20 CAS-IS strains (with disruption of FKS1, SMI1, BCK1, CHS4, ERG5, TPO1, and ILM1) and 1 of 9 CAS-IR strains (with disruption of SLG1) demonstrated selective susceptibility to CAS . To further explore the importance of PKC in CAS susceptibility, the activity of the PKC inhibitor staurosporine in combination with CAS was tested against eight Aspergillus clinical isolates by the microdilution assay . Synergistic or synergistic-to-additive activities were found against all eight isolates by use of both MIC and minimum effective concentration endpoints.

Antimicrob Agents Chemother, 2004 Oct, 48(10), 3823 - 7
Intrapulmonary pharmacokinetics and pharmacodynamics of itraconazole and 14-hydroxyitraconazole at steady state; Conte JE Jr et al.; We determined the steady-state intrapulmonary pharmacokinetic and pharmacodynamic parameters of orally administered itraconazole (ITRA), 200 mg every 12 h (twice a day {b.i.d.}), on an empty stomach, for a total of 10 doses, in 26 healthy volunteers . Five subgroups each underwent standardized bronchoscopy and bronchoalveolar lavage (BAL) at 4, 8, 12, 16, and 24 h after administration of the last dose . ITRA and its main metabolite, 14-hydroxyitraconazole (OH-IT), were measured in plasma, BAL fluid, and alveolar cells (AC) using high-pressure liquid chromatography . Half-life and area under the concentration-time curves (AUC) in plasma, epithelial lining fluid (ELF), and AC were derived using noncompartmental analysis . ITRA and OH-IT maximum concentrations of drug (C(max)) (mean +/- standard deviation) in plasma, ELF, and AC were 2.1 +/- 0.8 and 3.3 +/- 1.0, 0.5 +/- 0.7 and 1.0 +/- 0.9, and 5.5 +/- 2.9 and 6.6 +/- 3.1 microg/ml, respectively . The ITRA and OH-IT AUC for plasma, ELF, and AC were 34.4 and 60.2, 7.4 and 18.9, and 101 and 134 microg . hr/ml . The ratio of the C(max) and the MIC at which 90% of the isolates were inhibited (MIC(90)), the AUC/MIC(90) ratio, and the percent dosing interval above MIC(90) for ITRA and OH-IT concentrations in AC were 1.1 and 3.2, 51 and 67, and 100 and 100%, respectively . Plasma, ELF, and AC concentrations of ITRA and OH-IT declined monoexponentially with half-lives of 23.1 and 37.2, 33.2 and 48.3, and 15.7 and 45.6 h, respectively . An oral dosing regimen of ITRA at 200 mg b.i.d . results in concentrations of ITRA and OH-ITRA in AC that are significantly greater than those in plasma or ELF and intrapulmonary pharmacodynamics that are favorable for the treatment of fungal respiratory infection.

Antimicrob Agents Chemother, 2004 Oct, 48(10), 3670 - 6
Pharmacodynamic functions: a multiparameter approach to the design of antibiotic treatment regimens; Regoes RR et al.; There is a complex quantitative relationship between the concentrations of antibiotics and the growth and death rates of bacteria . Despite this complexity, in most cases only a single pharmacodynamic parameter, the MIC of the drug, is employed for the rational development of antibiotic treatment regimens . In this report, we use a mathematical model based on a Hill function-which we call the pharmacodynamic function and which is related to previously published E(max) models-to describe the relationship between the bacterial net growth rates and the concentrations of antibiotics of five different classes: ampicillin, ciprofloxacin, tetracycline, streptomycin, and rifampin . Using Escherichia coli O18:K1:H7, we illustrate how precise estimates of the four parameters of the pharmacodynamic function can be obtained from in vitro time-kill data . We show that, in addition to their respective MICs, these antibiotics differ in the values of the other pharmacodynamic parameters . Using a computer simulation of antibiotic treatment in vivo, we demonstrate that, as a consequence of differences in pharmacodynamic parameters, such as the steepness of the Hill function and the minimum bacterial net growth rate attained at high antibiotic concentrations, there can be profound differences in the microbiological efficacy of antibiotics with identical MICs . We discuss the clinical implications and limitations of these results.

Korean J Gastroenterol, 2004 Sep, 44(3), 126 - 35
{Antibiotic resistance of Helicobacter pylori isolated from Korean patients in 2003}; Kim JM et al.; BACKGROUND/AIMS: Development of antibiotic resistance is a significant clinical problem in the eradication of H . pylori . To select an appropriate regimen, systematic information on antibiotic resistance is mandatory . Thus, we investigated the distribution of minimal inhibitory concentration (MIC) and evaluated the antibiotic resistance of H . pylori isolates from Korean patients in 2003 . METHODS: The susceptibility of 65 isolates obtained in 2003 to amoxicillin, clarithromycin, metronidazole, tetracycline, azithromycin, and ciprofloxacin were determined by agar dilution method . RESULTS: Resistance rates of H . pylori to amoxicillin, clarithromycin, metronidazole, tetracycline, azithromycin, and ciprofloxacin were 18.5%, 13.8%, 66.2%, 12.3%, 32.3%, and 33.8%, respectively . Multi-drug resistance rate of H . pylori was 47.7% . Especially, 6.2% of the H . pylori isolates were resistant to both amoxicillin and clarithromycin . In addition, resistance to amoxicillin and clarithromycin resulted in decreasing tendency of the eradication efficacy for H . pylori . CONCLUSIONS: These results indicate that the antibiotics used for H . pylori eradication show high resistance rates in Korea . Furthermore, continuous surveillance of antibiotic susceptibilities should be needed and further increases in antibiotic resistance would require susceptibility testing before treatment to maximize the efficacy of H . pylori treatment.

Microbiol Immunol, 2004, 48(9), 655 - 60
Application of monoclonal antibody, specific for intracellular Orientia tsutsugamushi, to immunofluorescent antibody test for determining antibiotic susceptibility; Kim MK et al.; The simple quantification of viable intracellular bacteria is important for the study of an obligate intracellular bacterium, Orientia tsutsugamushi . We applied a novel monoclonal antibody (M686-13)--specific for intracellular Orientia--to an immunofluorescent antibody (IFA) test for determining antibiotic susceptibility of O . tsutsugamushi . M686-13 did not react with Orientia that was inhibited by doxycycline, although bacterial particles still remained in the cells . This preferential staining of proliferating bacteria made the IFA test rapid and precise . Using this method, we could successfully measure the minimal inhibitory concentration (MIC) of a Korean strain of O . tsutsugamushi to doxycycline and clindamycin . This method may be used in other procedures to evaluate the growth of Orientia.

Br J Cancer, 2004 Oct 18, 91(8), 1495 - 9
Loss of nonclassical MHC molecules MIC-A/B expression during progression of uveal melanoma; Vetter CS et al.; Uveal melanoma differs from cutaneous melanoma with respect to aetiology, metastatic behaviour and immune biology . The notion that loss of classical MHC class I molecules in uveal melanoma lesions is associated with an improved prognosis suggests that NK cells act as the predominant cells responsible for immune surveillance of this tumour . Consequently, immune escape mechanisms of uveal melanoma should impair the innate immunity . To this end, expression of the ligand for the NK receptor NKG2D, that is, MIC-A/B was expressed by 50% of primary tumours, but none of the metastatic lesions . MIC+ tumours were characterised by a NKG2D+ infiltrate, which was absent in MIC- lesions subsequent to chemoimmune therapy . Strikingly, MIC-A/B expression in metastatic lesions was observed subsequent to chemotherapy with fotemustine in one case . In summary, MIC/NKG2D interactions seem to be involved in the immune surveillance of primary uveal melanomas, whereas for metastatic tumours this ligand/receptor system seems not to be relevant, thus, suggesting an immune selection of MIC negative tumour cells.

Cancer, 2004 Oct 1, 101(7), 1594 - 600
Aspergillus terreus: an emerging amphotericin B-resistant opportunistic mold in patients with hematologic malignancies; Hachem RY et al.; BACKGROUND: Invasive aspergillosis (IA) has emerged as a common cause of morbidity and mortality among immunocompromised patients . At The University of Texas M . D . Anderson Cancer Center (Houston, TX), Aspergillus terreus is second to A . fumigatus as the most common cause of IA . In the current study, the authors compared the risk factors and outcomes associated with IA caused by A . terreus and IA caused by A . fumigatus . METHODS: The authors retrospectively reviewed the medical records of 300 patients who received care at our institution between 1995 and 2001 and who had cultures that were positive for Aspergillus infection, including 90 patients whose cultures were positive for A . fumigatus and 70 patients whose cultures were positive for A . terreus . RESULTS: Thirty-two patients with IA caused by A . terreus and 33 patients with IA caused by A . fumigatus were evaluated . The two groups were comparable in terms of age, gender, and underlying disease . Leukemia was the most common underlying malignancy (84%) . More than 40% of patients in each group had undergone bone marrow transplantation . There was a trend toward a higher frequency of neutropenia among patients with IA caused by A . terreus (P = 0.12) . IA caused by A . terreus was considered to be nosocomial in origin significantly more frequently compared with IA caused by A . fumigatus (P = 0.03) . In vitro, A . terreus was found to be more resistant to amphotericin B (minimal inhibitory concentration {MIC90}, 4.0 microg/mL) than to antifungal therapy (MIC90, 1.0 Hg/mL) in the isolates that were tested (< 50% of all isolates) . The overall rate of response to antifungal therapy was 39% for patients with A . fumigatus infection, compared with 28% for patients with A . terreus infection (P = 0.43) . CONCLUSIONS: Despite the decreased in vitro susceptibility of A . terreus (relative to A . fumigatus) to amphotericin B, the two groups within the current patient population had comparably poor responses to amphotericin B preparation and somewhat improved responses to posaconazole . (c) 2004 American Cancer Society.

Br J Clin Pharmacol, 2004 Oct, 58(4), 345 - 51
Comparison of the pharmacokinetics of miconazole after administration via a bioadhesive slow release tablet and an oral gel to healthy male and female subjects; Cardot JM et al.; AIMS: The aim of this study was to compare salivary miconazole pharmacokinetics following once daily application of bioadhesive tablets (50 or 100 mg), vs the current treatment with a gel (3 times a day, 375 mg day(-1)) . METHODS: A three way cross over study was carried out in 18 healthy subjects (nine males, nine females) with a 1 week washout period between each treatment . Plasma and salivary pharmacokinetics of miconazole were assessed over a 24-h period . RESULTS: In all subjects the tablets gave higher and more prolonged salivary miconazole concentrations than the gel . Thus salivary miconazole AUC(0,24 h) was 37.2 times greater for the 100 mg tablet (90% confidence interval {CI} 22.9, 60.5) and 18.9 times greater for the 50 mg tablet (CI 11.7, 30.6) compared with the gel . Similarly, Cmax was 17.2 times greater (CI 11.8, 25.2) and 7.8 times greater (CI 5.3, 11.4) for the 100 mg tablet and 50 mg tablet, respectively . Comparison of the 100 mg and 50 mg tablets gave ratios of 2.2 and 2.0 for Cmax and AUC(0,24 h), respectively (CI 1.5, 3.2 and 1.2, 3.2) . The mean time that salivary miconazole concentrations were above 0.4 micro g ml(-1) (the concentration reached 3 h after application of the oral gel according to published data) or above 1.0 microg ml(-1) (the MIC of some Candida species) was greater for both bioadhesive tablets than for the oral gel (10-14 h vs 1.5 h and 7 h vs 0.6 h) . Only 19 plasma samples from eight subjects had concentrations of miconazole above 0.4 micro g ml(-1) . Ten of these were taken from five subjects after administration of the gel and nine from three subjects after administration of the tablets . CONCLUSIONS: These data strongly support the further development of miconazole bioadhesive tablets as a sustained release formulation leading to improved antifungal exposure in the buccal cavity . A single daily application should improve compliance, whereas the low systemic absorption of miconazole will alleviate concerns regarding drug interactions and adverse effects .

Orv Hetil, 2004 Jul 11, 145(28), 1467 - 71
{The comparison of in vivo and in vitro elution of gentamycin from bone cement}; Balint L et al.; INTRODUCTION: The authors report their in vivo and in vitro results of the elution characteristics of gentamycin sulfate from bone cement, which is the most commonly used way of local antibiotic prophylaxis in Europe in the field of orthopedic surgery . AIM: The aim of this study was to investigate the elution of the gentamycin sulfate from bone cement, describe the dynamics of the emission in time and evaluate the relationship between the minimal inhibitory concentration (MIC) and the eluted concentration of the antibiotic . METHODS: The in vivo investigation samples were taken from 9 patient from drain fluids to evaluate the eluted antibiotic concentration by fluorescent polarisation immunoassay method (FPI) . The in vitro emission-dynamics of two different bone cement-Gentamycin sulphate complex were analysed by plate diffusion method during one-year period after mixing . RESULTS: Their results showed that 24 hours after the operation the gentamycin concentrations in the drain fluid taken from around the endoprosthesis implanted with Palacos-R bone cement diminished, yet remained above the MIC level . High but rapidly decreasing antibiotic level was detected by the in vitro method within the first week, reading an almost steadily low concentration by the end of the first month . Surprisingly, after one year it was still possible to demonstrate the inhibitory effect of the drug from both tested types of cements . CONCLUSIONS: It is concluded that the gentamycin is able to elute from the bone cement in useful concentration after the implantation of endoprosthesis . However, this in vitro method is a useful and reproducible technique for the measurement of the efficacy of antibiotic emission from bone cement, the conversation of the results to the in vivo remains to be obscured . Nevertheless, the usage of local antibiotic prophylaxis seems to be useful during orthopaedic major intervention.

J Infect Chemother, 2004 Aug, 10(4), 216 - 9
In vitro antifungal activity of luliconazole (NND-502), a novel imidazole antifungal agent; Uchida K et al.; The in vitro activity of luliconazole (NND-502), a novel imidazole antifungal agent, against dermatophytes and several other groups of medically important fungi including the rare causative agents of dermatomycoses, was studied . The luliconazole susceptibility tests were performed with a total of 58 fungal strains of 23 species of fungi grouped into dermatophytes, dematiaceous fungi, hyaline hyphomycetes, yeastlike fungi, and zygomycetes using a broth microdilution method with RPMI 1640 medium . The minimum inhibitory concentration (MIC) values for luliconazole were compared with those of three reference drugs, lanoconazole (LCZ), bifonazole (BFZ), and terbinafine (TBF), all of which have been popular for the topical treatment of dermatophytosis, cutaneous candidiasis, and other superficial fungal infections in Japan . Luliconazole inhibited growth of all filamentous fungi except zygomycetes at low concentrations (MIC, < or =0.004-0.125 microg/ml), with dermatophytes being most susceptible (MIC, < or =0.004-0.008 microg/ml) . The susceptibility of these filamentous fungi to luliconazole was almost equal to that to LCZ, and surpassed TBF and BFZ, although to a lesser extent; yeastlike fungi were also susceptible to luliconazole (MIC, 0.125-4 microg/ml) . Again the anti-yeastlike fungi activity of luliconazole was at the same level as LCZ and was greater than that of BFZ and TBF . In contrast to BFZ and TBF, however, luliconazole and LCZ were virtually inactive against zygomycetes.

Int J Gynecol Cancer, 2004 Sep-Oct, 14(5), 911 - 20
Estrogen receptor, progesterone receptor, and bcl-2 are markers with prognostic significance in CIN III; Fonseca-Moutinho JA et al.; There are no known biological markers or technologies to predict the natural history of an individual CIN III . The probability of progression is considered greater with the persistence of high-risk human papillomavirus (HPV) infection and age . p53 polymorphism has been associated with cervical carcinogenesis . Hormone-induced cervical cancer is mediated by estrogen receptor (ER) and progesterone receptor (PR) . In cervical cancer, increased bcl-2 and Bax immunoreactivity is generally associated with a better prognosis . The purpose of this study was to evaluate the value of HPV 16 and HPV 18 typing and p53 codon polymorphism genotyping by polymerase chain reaction and ER, PR, bcl-2, and Bax expression by immunohistochemistry in predicting the CIN III clinical behavior of CIN III lesions . We studied the expression of these prognostic factors in the CIN III adjacent to squamous cell microinvasive carcinomas of the cervix (MIC) from 29 patients with FIGO stage IA1 cervical cancer and in 25 patients with CIN III and no documented focus of invasion . In the MIC group, only the CIN III was considered at least 2 mm away from the microinvasive complex . The ER, PR, bcl-2, and Bax immunoreactivity was scored as positive (>10% staining cells) and negative (<10% staining cells) . No significant difference was observed between MIC and CIN III group concerning HPV infection and p53 polymorphism . The ER, PR, bcl-2, and Bax immunohistochemical expression was stronger and more frequent in the CIN III group . After multivariable analysis, coexpression of ER, PR, and bcl-2 was the only independent factor in defining low risk of progression for CIN III . Our study suggests that coexpression of ER, PR, and bcl-2 may be a useful tool in identifying the CIN III lesions with low risk of progression to cervical cancer.

Helicobacter, 2004 Oct, 9(5), 422 - 8
Selective and effective bactericidal activity of the cobalt (II) cation against Helicobacter pylori; Bruggraber SF et al.; BACKGROUND: Although the anti-Helicobacter pylori activity of bismuth is well established, the therapeutic potential of other metal ions against the organism is not known . MATERIALS AND METHODS: We measured the minimum inhibitory concentrations of a series of metal ions, including several cobalt (II) compounds against four type strains and seven clinical isolates of H . pylori using three standard broth culture media and a defined medium . Other intestinal bacteria were also investigated for specificity of action . RESULTS: Cobalt chloride had marked activity against H . pylori (minimum inhibitory concentration range was 0.03-1.0 mg/l) . The effect was specific because other transition metals had no effect and other intestinal bacteria were not affected by cobalt chloride . Activity was attributable to free cobalt ions as ligands inhibited activity in proportion to their affinity for the ions . Inhibition of cobalt activity was also observed in the presence of nickel, in a dose dependent fashion . However, cobalt activity was not directed towards the nickel-dependent urease enzyme because its effect was similar in wild-type and urease negative mutant strains of H . pylori . Finally, the viability of H . pylori was reduced at the same rate with 2 mg/l cobalt as with 1 mg/l amoxicillin . CONCLUSIONS: Cobalt competes for nickel in its acquisition by H . pylori, but mediates toxicity in a nonurease dependent fashion . As cobalt MIC is similar to some antibiotics and 10 to a hundred times lower than for bismuth, cobalt may represent an effective form of therapy for H . pylori infection.

Helicobacter, 2004 Oct, 9(5), 400 - 7
The rdxA gene plays a more major role than frxA gene mutation in high-level metronidazole resistance of Helicobacter pylori in Taiwan; Yang YJ et al.; BACKGROUND: Metronidazole-resistant H . pylori associating with mutations of rdxA or frxA is still a debated topic . This study investigates whether rdxA and frxA mutations of H . pylori accounted for the high MIC value (>/= 64 micro g/ml) of metronidazole (Mtz) . MATERIAL AND METHODS: From 126 clinical H . pylori isolates, we examined 14 Mtz-sensitive, 18 Mtz-resistant H . pylori, and eight pairs of Mtz-sensitive and Mtz-resistant colonies simultaneously present within a single gastric biopsy . The paired strains from one single biopsy were proven identical by PCR-RFLP . MICs of Mtz were checked by the E-test and agar dilution method . The mutations of rdxA and frxA sequencing were matched with the Mtz-susceptible ATCC 26695 and J99 . RESULTS: There were 89% (16/18) of Mtz-resistant isolates with mutation of RdxA . Half of the 14 Mtz-sensitive strains, all without mutation of RdxA, still contained truncation of FrxA . Within the paired isolates from a single biopsy, rdxA mutation (86%) was more common than frxA mutation (43%) in those isolates with high-level Mtz-resistant H . pylori . RdxA truncation was more prevalent in Mtz-resistant strains with high MICs than in those with low to moderate MICs (75% vs . 20%, p =.01, OR: 12, 95% CI: 1.8-81.7) . CONCLUSION: Mutations in the rdxA gene rather than the frxA gene generally determine a high MIC level of Mtz-resistant H . pylori in Taiwan.

Immunity, 2004 Sep, 21(3), 367 - 77
A direct role for NKG2D/MICA interaction in villous atrophy during celiac disease; Hue S et al.; MICA molecules interact with the NKG2D-activating receptor on human NK and CD8 T cells . We investigated the participation of the MICA/NKG2D pathway in the destruction of intestinal epithelium by intraepithelial T lymphocytes (IEL) in Celiac disease and its premalignant complication, refractory sprue . We show that MICA is strongly expressed at epithelial cell surface in patients with active disease and is induced by gliadin or its p31-49 derived peptide upon in vitro challenge, an effect relayed by IL-15 . This triggers direct activation and costimulation of IEL through engagement of NKG2D, leading to an innate-like cytotoxicity toward epithelial targets and enhanced TCR-dependent CD8 T cell-mediated adaptive response . Villous atrophy in Celiac disease might thus be ascribed to an IEL-mediated damage to enterocytes involving NKG2D/MICA interaction after gliadin-induced expression of MICA on gut epithelium . This supports a key role for MIC/NKG2D in the activation of intraepithelial immunity in response to danger.

Immunity, 2004 Sep, 21(3), 357 - 66
Coordinated induction by IL15 of a TCR-independent NKG2D signaling pathway converts CTL into lymphokine-activated killer cells in celiac disease; Meresse B et al.; A major function of NKG2D linking innate and adaptive immunity is to upregulate antigen-specific CTL-mediated cytotoxicity in tissues expressing stress-induced NKG2D ligands, such as MIC, by coactivating TCR signaling . Here, we show that, under conditions of dysregulated IL15 expression in vivo in patients with celiac disease and in vitro in healthy individuals, multiple steps of the NKG2D/DAP10 signaling pathway leading to ERK and JNK activation are coordinately primed to activate direct cytolytic function independent of TCR specificity in effector CD8 T cells . These findings may not only explain previous reports of transformation of CTL into NK-like "lymphokine-activated killers" (LAK cells) under high doses of IL2 (a substitute for IL15) but may also have significant implications for understanding and treating immunopathological diseases.

Mol Gen Mikrobiol Virusol, 2004, (3), 3 - 7
{Analysis of point mutations in the ygeD, gyrA and parC genes in fluoroquinolones resistant clinical isolates of Chlamydia trachomatis}; Misiurina OIu et al.; Resistance of 14 clinical isolates of C . trachomatis to fluoroquinolones, i.e . of ciprofloxacin, pefloxaxin and ofloxacin, was assayed . Three isolates with a high resistance degree to all 3 drugs (MIC equal or above 64 microg/ml) were detected . MIC was found to be equal to or below 4 microg/ml for 3 isolates . The remaining isolates had an intermediate resistance level . The nucleotide sequence was established for the Quinolone-Resistance Determining Region (QRDR) genes coding the DNA-gyrase subunit A (gyrA) and DNA-topoisomerase IV subunit C (parC) as well as for the 3'-region of ygeD coding, presumably, the efflux protein . In none of the isolates, the gyrA and gyrC QRDR differed from the corresponding regions in the published C . trachomatis genome sequence . Several silent mutations and mutations resulting in amino acid substitutions were observed in the ygeD 3' region of 2 isolates resistant to high FQ concentrations and in 1 isolate with the intermediate resistance level.

Can J Vet Res, 2004 Jul, 68(3), 229 - 31
Molecular basis of quinolone resistance in Escherichia coli from wild birds; Jimenez Gomez PA et al.; Nine quinolone resistant (minimal inhibitory concentration {MIC} was > 32 microg/mL for nalidixic acid, > 1 microg/mL for ciprofloxacin) isolates of Escherichia coli have been found in wild birds with septicemia . All of the isolates were aerobactin positive . The mechanisms of resistance were characterised by sequencing the quinolone resistance-determining region (QRDR) of the gyrA, gyrB, parC, and parE genes . Sequence analysis of the gyrA gene in all isolates identified only 1 nucleotide substitution at codon Serine-83 for Leucine-83 . Sequence analysis of the gyrB, parC, and parE QRDR genes revealed no mutations in any of the isolates . This study was conducted to determine the importance of these genes in the susceptibility of E . coli strains isolated from wild birds to quinolones.

Chemosphere, 2004 Nov, 57(6), 439 - 45
Evaluation of solubilizing ability of humic aggregate basing on the phase-separation model; Terashima M et al.; Solubilizing abilities of aggregates of humic acid (HA) to chlorinated benzenes (CBs) were investigated by means of the apparent water solubility enhancement . Both the water solubilities of 1,4-dichlorobenzene (DCB) and 1,2,4,5-tetrachlorobenzene (TeCB) linearly increased with increasing concentration of HA above the critical micelle concentration (CMC) . Such solubilization behavior of CBs for HA was compatible with those for sodium dodecyl sulfate (SDS) . These results indicate that the solubilization of CBs in the aqueous solution of HA above the CMC can be interpreted on the basis of the phase-separation model . Thus, the partition coefficients (K(mic)) of CBs between water and HA aggregate phases were calculated by assuming this model . The fact that the K(mic) value increased with increasing K(ow) of CBs supported the partition into the HA aggregate phase by hydrophobic interaction . The estimated K(mic) values of DCB were not dependent on the solution pH . Both K(mic) values of DCB and TeCB for the HA aggregate were found to be 4-5-fold lower than those of SDS.

J Mater Sci Mater Med, 2000 Jun, 11(6), 393 - 7
Controlled release systems based on poly(lactic acid) . An in vitro and in vivo study; Andreopoulos AG et al.; A new biodegradable delivery system based on poly(lactic acid) has been formulated, with potential applications in sustained antibiotic release against bone infection . The in vitro release of a new quinolone (pefloxacin) from low molecular weight poly(D,L-lactic acid) Mw = 2x10(3) lasted for 56 d whereas the in vivo delivery lasted 33 d . In both cases, the release rate is controlled by the drug diffusion and the polymer degradation, which seems to be the predominant factor . For the release experiments, discs were prepared from poly (D,L-lactide) Mw = 2x10(4) with drug loadings of 2% and 10% w/w . It was concluded that pefloxacin concentration remains higher than the Minimum Inhibitory Concentration (MIC) against the major causative bacteria of bone infection . The results indicate that the two different types of poly(lactic acid) can be used effectively in an implantable antibiotic release system .

Indian J Med Res, 2004 Aug, 120(2), 100 - 5
Correlation of mutations detected by INNO-LiPA with levels of rifampicin resistance in Mycobacterium tuberculosis; Srivastava K et al.; BACKGROUND & OBJECTIVES: Due to emergence of drug resistance in Mycobacterium tuberculosis, there is a need to have accurate and rapid methods for detection of drug resistance to important drugs like rifampicin . The present study was aimed at evaluation of a commercially available INNO-LiPA assay, for the detection of mutation in rpoB gene region of M . tuberculosis and correlate these mutations with levels of rifampicin resistance for assessing their clinical relevance . METHODS: Fifty five well-characterized isolates of M . tuberculosis deposited from various regions of India in Mycobacterial Repository Centre at the CJILOMD, Agra were subjected to susceptibility testing for rifampicin at various concentrations of drug viz., 10, 40, 64, 128 microg/ml on Lowenstein- Jensen (LJ) medium . rpoB gene fragment (260 bp) was amplified using Rif-TB amplification kit and after hybridization, detection was done by using INNO-LiPA Rif TB kit . RESULTS: The rpoB gene could be amplified from DNA extracted from all the 55 culture isolates and showed clear hybridization pattern with M . tuberculosis complex specific probes on LiPA strips . Mutations detected were correlated with degree of rifampicin resistance . All the sensitive isolates (identified by MIC) were identified as rifampicin sensitive (100%) by INNO-LiPA as they exhibit positive for wild type 'S' probes and negative for 'R' probes . Two of the 5 isolates, resistant at 10 microg/ml and 40 microg/ml had either D516V, H526Y mutations or unknown mutations . Thirty (85.71%) isolates resistant at clinically relevant levels (64,128microg/ml) exhibited double, triple or more 'R' type mutations (R(2(D516V)), R(4a(H526Y)), R(4b(H526D)), R(5(S531L))) as well as unknown mutations present at 'S' probes region whereas remaining isolates did not show any mutation by this method . This method could identify with definitiveness 60 per cent ( 21/35) isolates as rifampicin resistant as mutations observed in others were also present in isolates with low levels of resistance . INTERPRETATION & CONCLUSION: The results indicate that INNO-LiPA Rif TB test is a rapid and easy to use method for detection of mutations associated with rifampicin resistance in M . tuberculosis . However, as some of these mutations are also present in isolates with low degree of resistance which are still microbiologically sensitive to rifampicin, there is a need to improve this assay by exclusion of some of the current probes and inclusion of more probes.

J Antimicrob Chemother, 2004 Oct, 54(4), 755 - 60 Epub 2004 Sep 03.
Activity of capuramycin analogues against Mycobacterium tuberculosis, Mycobacterium avium and Mycobacterium intracellulare in vitro and in vivo; Koga T et al.; OBJECTIVES: The antimycobacterial activities of RS-112997, RS-124922 and RS-118641, three capuramycin analogues that inhibit phospho-N-acetylmuramyl-pentapeptide translocase, were tested against clinical isolates of Mycobacterium tuberculosis, Mycobacterium avium and Mycobacterium intracellulare . METHODS AND RESULTS: MICs were determined by the broth microdilution method using a modified Middlebrook 7H9 broth . RS-118641 was the most potent compound overall . The MIC50/90 (mg/L) results for RS-118641 were: M . tuberculosis, 1/2; multidrug-resistant (MDR) M . tuberculosis, 0.5/2; M . avium, 4/8; and M . intracellulare, 0.06/0.5 . No statistically significant differences in MIC distributions were observed between non-MDR and MDR M . tuberculosis for any of the capuramycin analogues tested . In order to evaluate the therapeutic efficacy of RS-112997 and RS-124922 in a murine lung model of tuberculosis, both compounds were administered intranasally at 0.1 or 1 mg/mouse/day for 12 days . The mycobacterial load in the lungs was significantly lower in all treatment groups than in the untreated controls . Additional experiments were performed to evaluate the therapeutic efficacy of the three compounds against the M . intracellulare infection in mice . All compounds were administered intranasally at 0.1 mg/mouse/day for 21 days . The mycobacterial load in the lungs was significantly lower in all treatment groups than in the untreated controls . CONCLUSIONS: These results suggest that capuramycin analogues exhibit strong antimycobacterial potential and should be considered for further evaluation in the treatment of M . tuberculosis and M . avium-M . intracellulare complex infections in humans.

Farmaco, 2004 Sep, 59(9), 673 - 8
1-Phenyl-3-toluyl-4-{ortho-1'-(N-ethyl-2'-methylpropylamine)}phenylpyrazole, synthesis and evaluation of the in vitro antifungal activity against Botrytis cinerea and Fusarium oxysporum; Dardari Z et al.; A novel antifungal pyrazole derivative was synthesized . Designated 1-phenyl-3-toluyl-4-{ortho-1'-(N-ethyl-2'-methylpropylamine)}phenylpyrazole, the compound exerted an antifungal effect toward Botrytis cinerea and Fusarium oxysporum . In fact, our results clearly show that mycelial growth and conidial germination of both fungi were blocked by the compound . Indeed, a 96-well microbioassay procedure was used for fast and easy evaluation of minimal inhibitory concentration (MIC) . The MIC values for B . cinerea and F . oxysporum were 25 and 36 microg/ml, respectively.

Eur J Med Chem, 2004 Sep, 39(9), 805 - 14
Ring-substituted imidazoles as a new class of anti-tuberculosis agents; Gupta P et al.; We describe in vitro anti-Mycobacterium tuberculosis activities of ring-substituted-1H-imidazole-4-carboxylic acid derivatives (1-6), and 3-(2-alkyl-1H-imidazol-4-yl)-propionic acid derivatives (7-13) against drug-sensitive and drug-resistant M . tuberculosis H37Rv strains . The most effective analogues, 2f (R=R(1)=c-C(5)H(9)), and 2h (R=R(1)=c-C(6)H(11)) have produced >90% inhibition at a concentration of <6.25 microg/ml in the drug-sensitive screen . Upon further evaluation against drug-resistant strains, both analogues 2f and 2h produced an MIC value of 25.0 microg/ml . The observation of significant anti-tuberculosis activity in some of these analogues describes the discovery of novel ring-substituted-1H-imidazole-4-carboxylic acid ethyl esters as a new class of anti-tuberculosis agents.

Antimicrob Agents Chemother, 2004 Sep, 48(9), 3586 - 90
Genetic and culture-based approaches for detecting macrolide resistance in Chlamydia pneumoniae; Riska PF et al.; Three clinical Chlamydia pneumoniae isolates for which the MIC of azithromycin increased after treatment were investigated for genetic evidence of macrolide resistance . Attempts to induce antibiotic resistance in vitro were made . No genetic mechanism was identified for the phenotypic change in these C . pneumoniae isolates . No macrolide resistance was obtained in vitro.

Antimicrob Agents Chemother, 2004 Sep, 48(9), 3567 - 9
T2182C mutation in 23S rRNA is associated with clarithromycin resistance in Helicobacter pylori isolates obtained in Bangladesh; Khan R et al.; Twelve clarithromycin-resistant (MIC, > or = 1 microg/ml) Helicobacter pylori isolates were analyzed for point mutations in the 23S rRNA gene . Sequence analysis of all of the resistant isolates revealed a T-to-C transition mutation at position 2182 . Transformation experiments confirmed that a single T-to-C transition mutation at position 2182 is associated with clarithromycin resistance.

Antimicrob Agents Chemother, 2004 Sep, 48(9), 3508 - 15
Steady-state plasma and intrapulmonary pharmacokinetics and pharmacodynamics of cethromycin; Conte JE Jr et al.; The objective of this study was to determine the steady-state plasma and intrapulmonary pharmacokinetic parameters of orally administered cethromycin in healthy volunteers . The study design included administering 150 or 300 mg of cethromycin once daily to 25 or 35 healthy adult subjects, respectively, for a total of five doses . Standardized and timed bronchoalveolar lavage (BAL) was performed after the last dose . Blood was obtained for drug assay prior to the first and last dose, at multiple time points following the last dose, and at the time of BAL . Cethromycin was measured in plasma, BAL, and alveolar cell (AC) by using a combined high-performance liquid chromatography-mass spectrometric technique . Plasma, epithelial lining fluid (ELF), and AC pharmacokinetics were derived by noncompartmental methods . C(max)/90% minimum inhibitory concentration (MIC(90)) ratios, area under the concentration-time curve (AUC)/MIC(90) ratios, intrapulmonary drug exposure ratios, and percent time above MIC(90) during the dosing interval (%T > MIC(90)) were calculated for recently reported respiratory pathogens . The kinetics were nonlinear, i.e., not proportional to dose . In the 150-mg-dose group, the C(max) (mean +/- standard deviations), AUC(0-24), and half-life for plasma were 0.181 +/- 0.084 microg/ml, 0.902 +/- 0.469 microg . h/ml, and 4.85 +/- 1.10 h, respectively; for ELF the values were 0.9 +/- 0.2 microg/ml, 11.4 microg . h/ml, and 6.43 h, respectively; for AC the values were 12.7 +/- 6.4 microg/ml, 160.8 microg . h/ml, and 10.0 h, respectively . In the 300-mg-dose group, the C(max) (mean +/- standard deviations), AUC(0-24), and half-life for plasma were 0.500 +/- 0.168 microg/ml, 3.067 +/- 1.205 microg . h/ml, and 4.94 +/- 0.66 h, respectively; for ELF the values were 2.7 +/- 2.0 microg/ml, 24.15 microg . h/ml, and 5.26 h, respectively; for AC the values were 55.4 +/- 38.7 microg/ml, 636.2 microg . h/ml, and 11.6 h, respectively . We concluded that the C(max)/MIC(90) ratios, AUC/MIC(90) ratios, %T > MIC(90) values, and extended plasma and intrapulmonary half-lives provide a pharmacokinetic rationale for once-daily administration and are favorable for the treatment of cethromycin-susceptible pulmonary infections.

Antimicrob Agents Chemother, 2004 Sep, 48(9), 3317 - 22
In vitro activities of voriconazole in combination with three other antifungal agents against Candida glabrata; Barchiesi F et al.; Candida glabrata has recently emerged as a significant pathogen involved in both superficial and deep-seated infections . In the present study, a checkerboard broth microdilution method was performed to investigate the in vitro activities of voriconazole (VOR) in combination with terbinafine (TRB), amphotericin B (AMB), and flucytosine (5FC) against 20 clinical isolates of C . glabrata . Synergy, defined as a fractional inhibitory concentration (FIC) index of < or = 0.50, was observed in 75% of VOR-TRB, 10% of VOR-AMB, and 5% of VOR-5FC interactions . None of these combinations yielded antagonistic interactions (FIC index > 4) . When synergy was not achieved, there was still a decrease in the MIC of one or both drugs used in the combination . In particular, the MICs were reduced to < or = 1.0 microg/ml as a result of the combination for all isolates for which the AMB MIC at the baseline was > or = 2.0 microg/ml . By a disk diffusion assay, the halo diameters produced by antifungal agents in combination were greater that those produced by each drug alone . Finally, killing curves showed that VOR-AMB exhibited synergistic interactions, while VOR-5FC sustained fungicidal activities against C . glabrata . These studies demonstrate that the in vitro activity of VOR against this important yeast pathogen can be enhanced upon combination with other drugs that have different modes of action or that target a different step in the ergosterol pathway . Further studies are warranted to elucidate the potential beneficial effects of such combination regimens in vivo.

Antimicrob Agents Chemother, 2004 Sep, 48(9), 3279 - 83
In vitro activity of caspofungin combined with sulfamethoxazole against clinical isolates of Aspergillus spp; Yekutiel A et al.; Caspofungin (CAS) inhibits fungal cell wall synthesis . Sulfamethoxazole (SMX) inhibits folate biosynthesis and is active in vitro against Aspergillus spp . We studied the activities of the combination of CAS and SMX against 31 Aspergillus isolates and compared them with that of SMX combined with amphotericin B (AMB) or itraconazole (ITC) . MICs and minimal effective concentrations (MECs) were determined by the NCCLS broth microdilution method . With MIC endpoints, the combination of SMX and CAS showed synergy or synergy to additivity against 29 of 31 isolates . With MEC endpoints, synergy to additivity was found against 12 of 31 isolates and indifference was displayed against the rest of them . SMX in combination with AMB or ITC was not truly synergistic, while synergy to additivity was found for SMX-AMB and SMX-ITC against 17 of 31 and 3 of 12 isolates, respectively . No antagonism was found with any of the drug combinations . Further analysis of the synergy of CAS and SMX was performed by detailed measurement of hyphal length by microscopy and time-dependent 2,3-bis(2-methoxy-4-nitro-5-{(sulfenylamino)carbonyl}-2H-tetrazolium hydroxide (XTT)-based hyphal damage experiments . With MEC endpoints, the combination of CAS and SMX was characterized by a greater than 50% decrease in hyphal length compared to the hyphal lengths achieved with double the concentration of each drug alone . The XTT-based hyphal damage studies showed a statistically significant (P < 0.05) reduction in viability with CAS and SMX in combination compared to the viabilities achieved with double the concentration of each drug alone . These findings support the synergy results found by using MIC endpoints and suggest that visual MEC measurements may not be sufficient to identify the synergistic interactions seen by more sensitive, quantitative methods . Animal models are required to validate the significance of the synergy of CAS and SMX against Aspergillus spp . observed in vitro.

Int J Antimicrob Agents, 2004 Sep, 24(3), 286 - 9
Trypanocidal effect of alpha',beta'-epoxyketones indicates that trypanosomes are particularly sensitive to inhibitors of proteasome trypsin-like activity; Glenn RJ et al.; Previous studies have shown that the proteasome of Trypanosoma brucei is a candidate for novel chemotherapy of African sleeping sickness . In this study, two potent and highly selective alpha',beta'-epoxyketones peptide proteasome inhibitors, epoxomicin and YU101, have been tested for their trypanocidal activities in vitro using culture-adapted bloodstream forms of T . brucei . Both inhibitors displayed promising anti-trypanosomal activities with ED(50) and ED(90) values in the low to mid nanomolar range . Based on MIC values, epoxomicin exhibited a selectivity index approaching those of commercially available drugs . Enzymatic analyses of proteasomal peptidase activities revealed that, compared with mammalian cells, trypanosomes are particular sensitive to inhibition of the trypsin-like activity of the proteasome . In conclusion, the data suggests that proteasome inhibitors targeting the trypsin-like activity are the rational choice for future anti-trypanosomal drug development.

Eur J Pharm Sci, 2004 Sep, 23(1), 77 - 87
Effect of different sterilisation methods on the properties of bioadhesive powders and ocular minitablets, and clinical evaluation; Weyenberg W et al.; The purpose of this study was to evaluate the influence of gamma-irradiation and dry heat sterilisation on the properties of a bioadhesive powder mixture containing ciprofloxacin and its corresponding ocular minitablets . The molecular weight characteristics of drum dried waxy maize starch (DDWM), employed as major component of the bioadhesive formulation, the decay kinetics of radicals, the rheological properties of the bioadhesive polymers and the microbial activity of ciprofloxacin were studied . The influence of the different sterilisation methods on the characteristics of the ocular minitablets was investigated by measuring the crushing strength, the friability, and the in vitro release of ciprofloxacin from the minitablets . Finally, the clinical value of the selected sterilised minitablets was evaluated in seven healthy volunteers . Both sterilisation methods similarly affected the properties of the bioadhesive formulation by inducing stable radicals and decreasing the molecular weight of DDWM, although no changes in the microbiological activity of ciprofloxacin were measured . An obvious influence of both sterilisation methods was observed in the in vitro release study . The crushing strength and friability of the minitablets were not significantly influenced by gamma-irradiation . Based on these data, gamma-irradiation was more adequate as sterilisation method for the bioadhesive ocular minitablets than dry heat sterilisation, because it affected the least the physical properties of the minitablets . Therefore, the gamma-sterilised minitablets were selected for an in vivo evaluation in seven volunteers . The concentration of ciprofloxacin in the tear film remained above its MIC value for the most common ocular pathogens for at least 8 h . Consequently, the gamma-irradiated minitablets containing ciprofloxacin can be considered as a promising formulation to treat bacterial keratitis and conjunctivitis.

Presse Med, 2004 Jul 10, 33(12 Pt 2), 2S5 - 9
{Stakes, treatment strategies and progression of MRSA nosocomial pneumonia, especially pneumonia due to mechanical ventilation}; Wunderink RG; THE IMPORTANCE OF THE INITIAL TREATMENT: Many studies have shown excess mortality during acquired pneumonia with mechanical ventilation when the initial antibiotic treatment is inappropriate, even following subsequent adaptation of the latter . EFFICACY OF TREATMENT: From a clinical point of view, since the regression of the various signs appears after varying time lapses, it is not easy to judge within the first three days the efficacy of an antibiotic . From a microbiological point of view, the bacterial concentrations observed at the time of diagnosis decrease within the first two days, when the response to treatment is favorable . PROBLEMS WITH VANCOMYCIN: Treatment of reference in the case of gram+ germ infections, vancomycin currently fails in 40% of MRSA pneumonias acquired under mechanical ventilation . The probable reason for such failure is an insufficient local concentration, which does not exceed the minimal inhibiting concentration (MIC) of the germ . BETWEEN EFFICACY AND TOLERANCE: The increase in the MIC of vancomycin in the serum and the lungs during acute MRSA acquired under mechanical ventilation may provoke problems in tolerance, notably renal.

J Natl Cancer Inst, 2004 Aug 18, 96(16), 1248 - 54
H6D polymorphism in macrophage-inhibitory cytokine-1 gene associated with prostate cancer; Lindmark F et al.; BACKGROUND: Accumulating epidemiologic and molecular evidence suggest that inflammation is an important component in the etiology of prostate cancer . Macrophage-inhibitory cytokine-1 (MIC-1), a member of the transforming growth factor beta superfamily, is thought to play an important role in inflammation by regulating macrophage activity . We examined whether sequence variants in the MIC-1 gene are associated with the risk of prostate cancer . METHODS: The study population, a population-based case-control study in Sweden, consisted of 1383 prostate cancer case patients and 780 control subjects . From 94 of the control subjects, we constructed gene-specific haplotypes of MIC-1 and identified four haplotype-tagging single-nucleotide polymorphisms (SNPs): Exon1+25 (V9L), Exon1+142 (S48T), IVS1+1809, and Exon2+2423 (H6D) . All study subjects were genotyped for the four SNPs, and conditional logistic regression analysis was used to estimate odds ratios (ORs) with 95% confidence intervals (CIs) . RESULTS: A statistically significant difference (P =.006) in genotype frequency was observed for the nonsynonymous change H6D (histidine to aspartic acid at position 6) between prostate cancer patients and control subjects . Carriers of the GC genotype, which results in the H6D change, experienced a lower risk of sporadic prostate cancer (OR = 0.80, 95% CI = 0.66 to 0.97) and of familial prostate cancer (OR = 0.61, 95% CI = 0.42 to 0.89) than the CC genotype carriers . In the study population, the proportion of prostate cancer cases attributable to the CC genotype was 7.2% for sporadic cancer and 19.2% for familial cancer . None of the other SNPs or haplotypes was associated with prostate cancer . CONCLUSION: This study shows an association between a nonsynonymous change (H6D) in the MIC-1 gene and prostate cancer . This finding supports the hypothesis that genetic variation in the inflammatory process contributes to prostate cancer susceptibility.

J Clin Invest, 2004 Aug, 114(4), 560 - 8
Prevalent expression of the immunostimulatory MHC class I chain-related molecule is counteracted by shedding in prostate cancer; Wu JD et al.; The MHC class I chain-related molecules (MICs) have previously been shown to be induced on most epithelial tumor cells . Engagement of MIC by the activating immune receptor NKG2D triggers NK cells and augments antigen-specific CTL anti-tumor immunity . The MIC-NKG2D system was proposed to participate in epithelial tumor immune surveillance . Paradoxically, studies suggest that tumors may evade MIC-NKG2D-mediated immunity by MIC shedding-induced impairment of effector cell function . Here we demonstrate the first evidence to our knowledge of a significant correlation of MIC shedding and deficiency in NK cell function with the grade of disease in prostate cancer . MIC is widely expressed in prostate carcinoma . The presence of surface target MIC, however, is counteracted by shedding . A significant increase in serum levels of soluble MIC (sMIC) and deficiency in NK cell function was shown in patients with advanced cancer . Finally, the deficiency in NK cell function can be overcome by treatment with IL-2 or IL-15 in vitro . Our results suggest that (a) deficiency in MIC-NKG2D immune surveillance may contribute to prostate cancer progression, (b) sMIC may be a novel biomarker for prostate cancer, and (c) using cytokines to restore MIC-NKG2D-mediated immunity may have clinical significance for prostate cancer in cell-based adaptive immunotherapy.

Mycoses, 2004 Aug, 47(7), 284 - 7
In vitro activity of terbinafine and itraconazole against Aspergillus species isolated from otomycosis; Karaarslan A et al.; The minimum inhibitory concentrations (MIC, microg ml-1) of itraconazole and terbinafine against overall 34 Aspergillus isolates from the external ear canals with otomycosis have been determined with M38-P microdilution method suggested by National Committee for Clinical Laboratory Standards (NCCLS) . MIC intervals in 48 h determined by taking MIC-2 value of itraconazole (the lowest drug concentration causing 50% inhibition of visible fungal growth) and MIC-0 value of terbinafine (the lowest drug concentration causing 100% inhibition of visible fungal growth) as a basis have been found as follows: 0.125-1 and 0.06-0.5 microg ml-1 for A . niger (22 strains), 0.06-0.25 and 0.06-0.125 microg ml-1 for A . flavus (10 strains), 0.125 and 0.125-0.5 microg ml-1 for A . terreus (two strains) . It has been observed that both of the antifungal agents showed an in vitro activity against all Aspergillus species tested.

Science, 2004 Sep 3, 305(5689), 1442 - 4 Epub 2004 Aug 12.
Substructure in the circumstellar disk around the young star AU Microscopii; Liu MC; Keck adaptive optics imaging with a physical resolution of 0.4 astronomical units (AU) resolves the inner (15 to 80 AU) disk of AU Microscopii (AU Mic, GJ 803, HD 197481), the nearest known scattered light disk to Earth . The inner disk is asymmetric and possesses a sharp change in structure at 35 AU . The disk also shows spatially localized enhancements and deficits at 25- to 40-AU separations . The overall morphology points to the influence of unseen larger bodies and resembles structures expected from recent planet formation . AU Mic is coeval with the archetypical debris disk system beta Pictoris, and the similarities between their two disks point to synchronous disk evolution . Multiple indications of substructure appear to be common in circumstellar disks at an age of approximately 12 million years.

Scand J Infect Dis, 2004, 36(6-7), 431 - 4
Evaluation of restriction endonuclease analysis of BRO beta-lactamases in clinical and carrier isolates of Moraxella catarrhalis; Koseoglu O et al.; A rapid increase in the prevalance of beta-lactamase producing M . catarrhalis isolates has highlighted its pathogenic potential . In this study, we aimed to detect the BRO beta-lactamases of our clinical (n = 32) and carrier (n =32) strains of Moraxella catarrhalis and compare the relationship of the enzyme type in assesment of MIC results of the antibiotics tested . BRO beta-lactamases were differentiated by restriction endonuclease analysis . Antibiotic susceptibility was performed by the agar dilution method recommended by NCCLS (M7A5) . The clinical isolates produced 96.9%, whereas the carrier strains produced 90.6% beta-lactamase positivity by the restriction enzyme analysis . BRO-1 was isolated as 90.6% (n =29) while the BRO-2 and non-beta-lactamase producers (NBLP) were isolated as 6.3% (n =2) and 3.1% (n =1) respectively among clinical isolates . The rate of BRO-1 in the carrier strains was 75.0% (n =24), BRO-2 was 15.6% (n =5) and NBLP was 9.4%, (n =3) . The beta-lactamase production with nitrocefin test was 96.9% (31/32) in clinical isolates and 90.6% (29/32) in carrier strains . M . catarrhalis needs a continous monitoring of antibiotic susceptibility; in this era restriction endonuclease analysis could be useful to screen BRO beta-lactamase genes.

Tissue Antigens, 2004 Sep, 64(3), 276 - 80
Eight novel MICB alleles, including a null allele, identified in gastric MALT lymphoma patients; Schroeder M et al.; MICA and MICB, as members of the major histocompatibility complex (MHC) class I-chain-related genes (MIC), encode stress-inducible glycoproteins that act as activating ligands for NKG2D and gammadelta T-cell receptor-bearing cells . We here describe the identification of eight novel MICB variants, including a null allele, which were identified in peripheral blood leukocytes of gastric MALT lymphoma patients . Only two of the novel alleles are characterized by point mutations, whereas the other variants display a recombination of known exonic MICB sequences that may be best explained by intragenic conversions . The novel MICB null allele is characterized by a Cytosin (C) deletion in a stretch of four Cs beginning from nucleotide 135 of exon 2 that leads to a premature stop codon (TGA) at codon 66.

Clin Microbiol Infect, 2004 Aug, 10(8), 757 - 60
First identification of an Escherichia coli clinical isolate producing both metallo-beta-lactamase VIM-2 and extended-spectrum beta-lactamase IBC-1; Galani I et al.; An Escherichia coli strain with decreased susceptibility to carbapenems was isolated from a hospitalised patient in Athens, Greece . The strain was resistant to all beta-lactams, including aztreonam, whereas the MIC of imipenem and meropenem was 0.5 mg/L . A positive EDTA-disk synergy test suggested the production of a metallo-beta-lactamase . PCR experiments revealed the presence of the bla(VIM-2), bla(IBC-1), and bla(TEM-1) genes . Resistance to beta-lactams was not transferable by conjugation . This is the first report of a clinical isolate of E . coli producing VIM-2, and the first report of the coexistence of bla(VIM-2) and bla(IBC-1) in a single clinical isolate.

Aliment Pharmacol Ther, 2004 Jul, 20 Suppl 1, 68 - 73
Second-line treatment of Helicobacter pylori infection after dilution agar methods and PCR-RFLP analysis; Masaoka T et al.; BACKGROUND: After unsuccessful first-line treatment of Helicobacter pylori infection, the percentage of clarithromycin-resistant strains has been reported as between 30% and 70% in Japan and other countries . A high prevalence of clarithromycin-resistant strains is reported to be associated with eradication failure . AIM: We examined antibiotic susceptibility testing using a combination of dilution agar methods with PCR-restriction fragment length polymorphism (RFLP) analysis . METHODS: We enrolled 41 patients in whom first-line treatment with LAC (lansoprazole, amoxycillin and clarithromycin) was unsuccessful . Endoscopic biopsied specimens were used to examine antibiotic susceptibility to clarithromycin by dilution agar methods . PCR-RFLP analysis was performed to determine the presence of point mutations, which are primarily responsible for resistance to clarithromycin . RESULTS: Clarithromycin-resistance rate after failure of the LAC regimen was 73.2% . Drug susceptibilities of three strains obtained by PCR-RFLP analysis were different from those by dilution agar methods . One strain with MIC values to clarithromycin of 0.05 micro g/mL had a point mutation, A2144G . This strain was not eradicated by repeating LAC, but was eradicated by substituting metronidazole for clarithromycin . CONCLUSIONS: Dilution agar methods should be combined with PCR-RFLP analysis before second-line eradication to increase the accuracy of clarithromycin-susceptibility testing and to improve eradication efficacy.

Aliment Pharmacol Ther, 2004 Jul, 20 Suppl 1, 62 - 7
High prevalence rate of Helicobacter pylori resistance to clarithromycin during long-term multiple antibiotic therapy for chronic respiratory disease caused by non-tuberculous mycobacteria; Kaneko F et al.; BACKGROUND: Helicobacter pylori resistance to clarithromycin, probably due to the frequent use of this antibiotic for the treatment of other diseases, is the greatest obstacle against its eradication . AIM: To clarify the prevalence of clarithromycin-resistant H . pylori in patients with non-tuberculous mycobacterial lung disease receiving multiple antibiotic treatment, including clarithromycin . METHODS: We enrolled 88 patients with non-tuberculous mycobacterial lung disease; 29 underwent upper gastrointestinal endoscopy for the diagnosis of H . pylori infection prior to treatment, and 60 underwent it during treatment . The diagnosis of H . pylori infection was confirmed by histological examination, urease test and microaerobic bacterial culture . The minimum inhibitory concentration of clarithromycin was determined and the DNA was analysed for each of the isolated H . pylori strains . RESULTS: Patients during the treatment had a high prevalence rate of clarithromycin-resistant H . pylori (100%) . Analysis of DNA of the clarithromycin-resistant H . pylori isolates revealed point mutations at A2142G or A2143G . Moreover, a linear correlation was found between the total cumulative dose of clarithromycin and the minimum inhibitory concentration . CONCLUSION: All patients with non-tuberculous mycobacterial lung disease being treated long-term with multiple antibiotics, including clarithromycin, harboured clarithromycin-resistant H . pylori in the stomach . Therefore, eradication of H . pylori before commencement of long-term therapy including clarithromycin should be recommended.

J Clin Microbiol, 2004 Aug, 42(8), 3607 - 12
Comparison of results of fluconazole disk diffusion testing for Candida species with results from a central reference laboratory in the ARTEMIS global antifungal surveillance program; Pfaller MA et al.; The accuracy of antifungal susceptibility tests is important for accurate resistance surveillance and for the clinical management of patients with serious infections . Our main objective was to compare the results of fluconazole disk diffusion testing of Candida spp . performed by ARTEMIS participating centers with disk diffusion and MIC results obtained by the central reference laboratory . A total of 2,949 isolates of Candida spp . were tested by NCCLS disk diffusion and reference broth microdilution methods in the central reference laboratory . These results were compared to the results of disk diffusion testing performed in the 54 participating centers . All tests were performed and interpreted following NCCLS recommendations . Overall categorical agreement between participant disk diffusion test results and reference laboratory MIC results was 87.4%, with 0.2% very major errors (VME) and 3.3% major errors (ME) . The categorical agreement between the disk diffusion test results obtained in the reference laboratory with the MIC test results was similar: 92.8% . Likewise, good agreement was observed between participant disk diffusion test results and reference laboratory disk diffusion test results: 90.4%, 0.4% VME, and 3.4% ME . The disk diffusion test was especially reliable in detecting those isolates of Candida spp . that were characterized as resistant by reference MIC testing . External quality assurance data obtained by surveillance programs such as the ARTEMIS Global Antifungal Surveillance Program ensure the generation of useful surveillance data and result in the continued improvement of antifungal susceptibility testing practices.

Drug Resist Updat, 2004 Jun, 7(3), 185 - 94
Antifungal pharmacokinetics and pharmacodynamics: understanding the implications for antifungal drug resistance; Andes D; Pharmacodynamics (PDs) describe the relationship between drug exposure and outcome . The drug exposures in these analyses are most commonly expressed in a variety of pharmacokinetic terms . The outcome of interest with anti-infective therapy is either microbiologic resolution or a clinical surrogate of treatment efficacy . An in vitro measure of drug potency, such as the minimum inhibitory concentration (MIC) is also frequently considered in this relationship . Examination of the relationships among drug pharmacokinetics, MIC, and efficacy has provided a framework for choice of antifungal drug and dose . These analyses provide a PD target for drug class/organism combinations . The PD target can be useful for defining the upper MIC limit for a drug-dosing regimen that would be expected to result in treatment efficacy . The PD target can be used to optimize dosing regimens and to aid in defining susceptibility breakpoints.

Pharmazie, 2004 Jul, 59(7), 568 - 70
A novel antifungal pyrrole derivative from Datura metel leaves; Dabur R et al.; Phytochemical investigation of the leaves of Datura metel Linn . led to the isolation of a new pyrrole derivative 1 which was characterised as 2beta-(3,4-dimethyl-2,5-dihydro-1H-pyrrol-2-yl)-1'-methylethyl pentanoate on the basis of spectral data analyses and chemical reactions . Compound 1 was endowed with antifungal activity and its MIC was found to be 87.5 microg/ml . Two proteins having molecular weights of 42 and 58 kD of Aspergillus fumigatus are potential targets for compound 1.

J AOAC Int, 2004 May-Jun, 87(3), 579 - 86
Relationships between LC retention, octanol-water partition coefficient, and fungistatic properties of 2-(2,4-dihydroxyphenyl)benzothiazoles; Matysiak J et al.; The retention behavior of newly synthesized compounds with antimycotic activity from the 2-(2,4-dihydroxyphenyl)benzothiazole group by high-performance liquid chromatography has been investigated . RP-18 stationary phase and methanol-acetate buffer aqueous mobile phases at pH 4 and 7.4 have been used . In the case of the mobile phase at pH 7.4, higher concentrations of water can be applied than at pH 4 . The studied compounds showed regular retention behavior, their log k values decreasing linearly with an increasing concentration of methanol in the mobile phase . On the basis of these relationships, the lipophilicity (log kw), specific hydrophobic surface area (S), and isocratic chromatographic hydrophobicity index (psi0) were determined . Similar log kw values and sensitivity to changes in the structure of compounds studied for both mobile phases have been found . Moderate correlations between the chromatographic parameters and the calculated octanol-water log P values were found . Finally, the lipophilicity parameters were compared with the fungistatic properties of compounds expressed by log MIC (minimum inhibitory concentration) values to find quantitative structure activity relationship equations.

Drug Metabol Drug Interact, 2004, 20(1-2), 101 - 8
In vitro susceptibility of Candida species isolated from cancer patients to some antifungal agents; Ozcelik B et al.; This study was undertaken to study the resistance of Candida species isolated from oropharyngeal swabs of cancer patients to ketoconazole (KET), fluconazole (FLU), amphotericin B (AmpB), and flucytosine (FCU) . The most common species identified was C . albicans, followed by C . tropicalis, C . glabrata, C . famata, C . krusei, C . kefyr, and C . gulliermondii . The minimum inhibitory concentration (MIC) of the antifungal agents was evaluated by RPMI 1640 medium with microdilution method . There were no C . albicans strains resistant to KET, FLU and AmpB . All Candida isolates were found highly susceptible to AmpB (MIC AmpB < 1 microg/ml), followed by KET (MIC KET < or =8 microg/ml), FLU (MIC FLU < or =8 microg/ml) and FCU (MIC FCU < or =4 microg/ml) . The main conclusion of this study is that prophylactic therapy planned according to typing and antifungal susceptibility will contribute to the prevention of invasive fungal infections in immunosuppressied oncology patients.

Antimicrob Agents Chemother, 2004 Aug, 48(8), 2951 - 7
Isoniazid pharmacokinetics-pharmacodynamics in an aerosol infection model of tuberculosis; Jayaram R et al.; Limited data exist on the pharmacokinetic-pharmacodynamic (PK-PD) parameters of the bactericidal activities of the available antimycobacterial drugs . We report on the PK-PD relationships for isoniazid . Isoniazid exhibited concentration (C)-dependent killing of Mycobacterium tuberculosis H37Rv in vitro, with a maximum reduction of 4 log10 CFU/ml . In these studies, 50% of the maximum effect was achieved at a C/MIC ratio of 0.5, and the maximum effect did not increase with exposure times of up to 21 days . Conversely, isoniazid produced less than a 0.5-log10 CFU/ml reduction in two different intracellular infection models (J774A.1 murine macrophages and whole human blood) . In a murine model of aerosol infection, isoniazid therapy for 6 days produced a reduction of 1.4 log10 CFU/lung . Dose fractionation studies demonstrated that the 24-h area under the concentration-time curve/MIC (r2 = 0.83) correlated best with the bactericidal efficacy, followed by the maximum concentration of drug in serum/MIC (r2 = 0.73).

J Vector Ecol, 2004 Jun, 29(1), 101 - 8
The in vitro effect of albendazole, ivermectin, diethylcarbamazine, and their combinations against infective third-stage larvae of nocturnally subperiodic Brugia malayi (Narathiwat strain): scanning electron microscopy; Tippawangkosol P et al.; Scanning electron microscopy (SEM) was employed to observe the effects of ivermectin (IVM), diethylcarbamazine (DEC), and albendazole (ALB) alone, and the drugs in combination (ALB+IVM and ALB+DEC) against infective third stage larvae (L3) of nocturnally subperiodic (NSP) Brugia malayi (Narathiwat strain) in vitro . IVM, at a concentration of 10(-4) M, killed L3 within 1-2 h . The SEM data showed damage to the L3 surface and loss of regular cuticular annulations . The cuticles were grooved in the middle region of the body . In comparison with normal L3 before treatment with IVM, the cuticular surface showed transversed striations with periodic annulations . The result demonstrated that IVM showed a larvicidal activity against L3 of NSP B . malayi cultivated in vitro . Compared with those larvae in the control group, the treated larvae had no morphological changes in the cuticular surface at the head, body, and tail regions after cultivation with all drugs alone, and in their combinations at a concentration of 10(-5) M for 7 d . In this system, and at that concentration, only the larvae cultured with ALB alone remained highly motile . Although no morphological changes had been observed by SEM, those drugs used alone (IVM and DEC) and in combinations (ALB+IVM and ALB+DEC), reduced larval motility throughout the experiments at a concentration of 10(-5) M . The minimum lethal concentration (MIC) of IVM against NSP B . malayi was 10(-4) M.

Eur J Clin Microbiol Infect Dis, 2004 Aug, 23(8), 619 - 24 Epub 2004 Jul 16.
In vitro activity of voriconazole and other antifungal agents against clinical isolates of Candida glabrata and Candida krusei; Drago M et al.; The antifungal susceptibility of 309 Candida glabrata and 63 Candida krusei clinical isolates was tested via the Sensititre YeastOne-3 system (Trek Diagnostic Systems, East Grinstead, UK) to compare the in vitro activity of voriconazole with that of five other antifungal agents (amphotericin B, fluconazole, itraconazole, ketoconazole, and flucytosine) . Voriconazole was highly active (MIC90, 0.5 microg/ml) against isolates of both species, including those for which the MICs of itraconazole and fluconazole were high (MIC90s of itraconazole, 2 microg/ml for C . glabrata and 0.5 microg/ml for C . krusei; MIC90s of fluconazole, 32 microg/ml for C . glabrata and 64 microg/ml for C . krusei) . Ketoconazole MIC90 values for both species were identical to those of voriconazole . The MIC90 of amphotericin B was similar for both species (0.125 microg/ml for C . glabrata and 0.25 microg/ml for C . krusei) . As expected, flucytosine was only moderately active against C . krusei isolates (MIC90, 16 microg/ml) but was highly active against C . glabrata isolates (MIC90, 0.03 microg/ml) . Potential cross-resistance within the azole class was noted for some strains of C . glabrata (5.5%) that presented high MIC values for all the azoles tested . In order to consider voriconazole a viable alternative to other triazoles for the treatment of infections caused by Candida species, susceptibility testing of all clinically significant isolates of C . glabrata and C . krusei is recommended because of the potential for azole cross-resistance . The Sensititre YeastOne-3 seems to be a suitable commercial tool for this purpose.

Zhonghua Jie He He Hu Xi Za Zhi, 2004 Jun, 27(6), 403 - 6
{A study on the active efflux mechanism in straphylococcus aureus resistant to quinolone}; Xu F et al.; OBJECTIVE: To explore the effect, the characteristic, and potential approaches to the active efflux mechanism in straphylococcus aureus (S . a) resistant to quinolones . METHODS: S . a standard strain ATCC25923 and clinical isolates susceptible to quinolone were inoculated onto MH agar containing ofloxacin at a concentration of 4 x MIC (in the presence or absence of 20 micro g of reserpine/ml) . Following incubation at 35 degrees C for 48 h, the inhibiting effect of reserpine on the occurrence of induced resistance was observed and the MIC of the induced resistant strain to ethidium bromide (EB), ofloxacin and ciprofloxacin (in the presence or absence of 20 micro g of reserpine/ml) was determined . The influence of reserpine to the MIC of induced resistant strains was also determined . The accumulation and loss of EB was determined based on the fact that EB's fluorescence can be strengthened when combined with DNA . Reserpine inhibition test was used to study the active efflux in clinical S . a resistant to quinolone . RESULTS: The active efflux mechanism in S . a resistant to second generation quinolones was confirmed by the reserpine's influence on the level of ethidium bromide (EB) in the cytoplasm of S . a . Reserpine reduced the 50 percent resistant rate to quinolone in induced resistant S . a, and decreased the MIC of induced resistant strains . Reserpine inhibited the active efflux of EB from the cytoplasm of S . a . CONCLUSIONS: Active efflux is an important mechanism in S . a resistant to quinolone . Reserpine can inhibit its active efflux mechanism, and has synergistic effect with quinolone, which hold therapeutic potential for S . a resistant to quinolones.

Planta Med, 2004 Jul, 70(7), 615 - 9
Inhibition of Helicobacter pylori adhesion to human gastric adenocarcinoma epithelial cells by acidic polysaccharides from Artemisia capillaris and Panax ginseng; Lee JH et al.; Helicobacter pylori specifically adheres to host cells, mainly based on carbohydrate-mediated cell-cell interactions . Previously, we investigated the anti-adhesive effect of polysaccharide fractions from Artemisia capillaris and Panax ginseng, using hemagglutination and enzyme-linked glycosorbent assays . In the present study, each active polysaccharide fraction was further purified, resulting in a single peak (fraction F2) using gel filtration FPLC, in which no protein content was detectable . Using scanning electron microscopy, we examined the inhibitory effects of these polysaccharides on the attachment of H . pylori to the human gastric adenocarcinoma epithelial cell line . The bacterial attachment to the cell line was inhibited by these polysaccharides in the range of the concentrations studied (0.2 - 2.8 mg/mL), showing their minimum inhibitory concentration at as low as 0.2 mg/mL . The bacterial binding was inhibited more effectively by P . ginseng polysaccharides, than by those from A . capillaris . The purified polysaccharides contain similar sugar compositions and have high amounts of uronic acids . Our results suggest that acidic carbohydrates may play an important role in the inhibitory activity on H . pylori adhesion to host cells and that our established purification protocol can be applied to obtain active acidic polysaccharides from plant sources.

Pediatr Infect Dis J, 2004 Jul, 23(7), 625 - 9
Ampicillin and penicillin concentration in serum and pleural fluid of hospitalized children with community-acquired pneumonia; Giachetto G et al.; BACKGROUND: Optimal therapeutic efficacy of beta-lactam antibiotics for treatment of pneumococcal pneumonia is thought to be associated with the serum concentration greater than the minimum inhibitory concentration for 40-50% of the interdose interval at site of infection . OBJECTIVE: Establish whether intravenous administration of ampicillin 400 mg/kg/day or penicillin 200,000 IU/kg/day in 6 divided doses reaches serum and or pleural concentrations above 4 microg/ml for at least 40% of the interdose interval . MATERIALS AND METHODS: Hospitalized healthy children 1 month-14 years old with community-acquired bacterial pneumonia and empyema were eligible . Blood samples were obtained 30 min (C1) and 3 h (C2) after an antibiotic dose . Pleural fluid samples were obtained 1 and 4 h after the same dose in which blood samples were obtained . The concentrations were measured by high performance liquid chromatography . RESULTS: The study included 17 patients treated with ampicillin and 13 treated with penicillin . For ampicillin, mean serum concentrations were C1 37.3 +/- 19 microg/ml and C2 11 +/- 10.2 microg/ml and mean pleural fluid concentrations were C1 25.8 +/- 9.9 microg/ml and C2 16.2 +/- 7.9 microg/ml . For penicillin, mean serum concentrations were C1 21.8 +/- 16.4 microg/ml and C2 23.9 +/- 3.4 microg/ml . Mean pleural fluid concentrations were C1 10.9 +/- 2.2 microg/ml and C2 7.7 +/- 3.4 microg/ml . In 8 of 30 patients, serum C2 was <4 microg/ml; in all of them serum concentrations were >4 microg/ml for >40% of the interdose interval . CONCLUSIONS: This study of the pharmacokinetics of beta-lactam antibiotics in children with bacterial pneumonia may help in the development of therapeutic guidelines for the treatment of pneumococcal pneumonia.

Bioorg Med Chem, 2004 Aug 1, 12(15), 4179 - 88
Synthesis and antimycobacterial activities of ring-substituted quinolinecarboxylic acid/ester analogues . Part 1; Vaitilingam B et al.; Structural optimization of recently discovered new chemical entity, 2,8-dicyclopentyl-4-methylquinoline (DCMQ; MIC= 6.25 microg/mL, M . tuberculosis H37Rv) resulted in the synthesis of four new series of ring-substituted quinolinecarboxylic acids/esters constituting 45 analogues . All new derivatives were evaluated for in vitro antimycobacterial activities against M . tuberculosis H37Rv . Certain ring-substituted-2-quinolinecarboxylic acid ester and ring-substituted-2-quinoline acetic acid ester analogues described herein showed moderate to good inhibitory activity . In particular, three analogues methyl 4,5-dicyclopentyl-2-quinolinecarboxylate (3b), methyl 4,8-dicyclopentyl-2-quinolinecarboxylate (3c) and ethyl 2-(2,8-dicyclopentyl-4-quinolyl)acetate (14g) exhibited excellent MIC values of 1.00, 2.00 and 4.00microg/mL, respectively . Results obtained indicate that substitution of the quinoline ring with dicyclopentyl substituent presumably enhances the antimycobacterial activities in the quinoline analogues described herein.

J Clin Microbiol, 2004 Jul, 42(7), 3142 - 6
Geographic variation in the susceptibilities of invasive isolates of Candida glabrata to seven systemically active antifungal agents: a global assessment from the ARTEMIS Antifungal Surveillance Program conducted in 2001 and 2002; Pfaller MA et al.; We examined the susceptibilities to amphotericin B, flucytosine, fluconazole, posaconazole, ravuconazole, voriconazole, and caspofungin of 601 invasive isolates of Candida glabrata and grouped the isolates by geographic location: North America (331 isolates), Latin America (58 isolates), Europe (135 isolates), and Asia-Pacific (77 isolates) . Caspofungin (MIC at which 90% of isolates tested are susceptible {MIC(90)}, 0.12 microg/ml; 100% of strains are susceptible {S} at a MIC of </=1 microg/ml) and flucytosine (MIC(90), 0.12 microg/ml; 99.2% S) were the most active agents in all geographic regions . Fluconazole susceptibility was highest in the Asia-Pacific region (80.5% S, 3.9% resistant {R}) and lowest in North America (64% S, 10.3% R) and Latin America (62.1% S, 3.4% R) . The extended-spectrum triazoles were most active in the Asia-Pacific region (90 to 96.1% S) and least active in North America (82.5 to 90.3% S) . All 46 isolates that were resistant to fluconazole were susceptible to caspofungin (MIC(90), 0.06 microg/ml) and flucytosine (MIC(90), 0.12 microg/ml) and exhibited variable cross-resistance to posaconazole, ravuconazole, and voriconazole.

J Clin Microbiol, 2004 Jul, 42(7), 2977 - 9
Intra- and interlaboratory study of a method for testing the antifungal susceptibilities of dermatophytes; Ghannoum MA et al.; The National Committee for Clinical Laboratory Standards (NCCLS) M38-A standard for the susceptibility testing of conidium-forming filamentous fungi does not explicitly address the testing of dermatophytes . This multicenter study, involving six laboratories, investigated the MIC reproducibility of seven antifungal agents tested against 25 dermatophyte isolates (5 blinded pairs of five dermatophyte species per site for a total of 300 tests), using the method of dermatophyte testing developed at the Center for Medical Mycology, Cleveland, Ohio . The dermatophytes tested included Trichophyton rubrum, Trichophyton mentagrophytes, Trichophyton tonsurans, Epidermophyton floccosum, and Microsporum canis . Seven antifungals with activity against dermatophytes were tested, including ciclopirox, fluconazole, griseofulvin, itraconazole, posaconazole, terbinafine, and voriconazole . Interlaboratory MICs for all isolates were in 92 to 100% agreement at a visual endpoint reading of 50% inhibition as compared to the growth control and 88 to 99% agreement at a visual endpoint reading of 80% inhibition as compared to the growth control . Intralaboratory MICs between blinded pairs were in 97% agreement at a visual endpoint reading of 50% inhibition as compared to the growth control and 96% agreement at a visual endpoint reading of 80% inhibition as compared to the growth control . Data from this study support consideration of this method as an amendment to the NCCLS M38-A standard for the testing of dermatophytes.

J Antimicrob Chemother, 2004 Aug, 54(2), 435 - 40 Epub 2004 Jul 08.
Helicobacter pylori antibiotic resistance patterns and genotypes in adult dyspeptic patients from a regional population in North Wales; Elviss NC et al.; OBJECTIVE: Surveillance data on Helicobacter pylori antibiotic susceptibilities in Wales are limited, despite resistance being a key factor in treatment failure . A single-centre survey was undertaken over 3 years to determine local antibiotic resistance rates on isolates from dyspeptic patients in Bangor, Gwynedd (North Wales) . METHODS: Susceptibilities were determined for 363 isolates by disc diffusion and the Etest . Isolates were also genotyped (cagA presence and vacA allelic types) . RESULTS: Overall in vitro resistance rates were 24% for metronidazole and 7% for clarithromycin, with 4% resistant to both antibiotics . Resistant strains typically had high MICs of >256 mg/L . Tetracycline resistance was identified in only one isolate whereas no isolates showed resistance to amoxicillin . There was a two-fold increase in resistance over the study period . No gender and age associations with resistance were detected . Resistant and susceptible isolates were genotypically diverse with respect to cagA/vacA type but the vacA s1m2 form was a feature of all clarithromycin-resistant isolates compared with 56% of the susceptible isolates . CONCLUSION: Although the overall antibiotic resistance rates of H . pylori from North Wales were low compared with many other regions in Europe, continued surveillance, particularly of high-level resistance (MIC >256 mg/L), is recommended to monitor the effects of the 'test and treat' strategy for H . pylori eradication.

Planta Med, 2004 Jun, 70(6), 566 - 8
In vitro anti-adhesive activity of an acidic polysaccharide from Panax ginseng on Porphyromonas gingivalis binding to erythrocytes; Lee JH et al.; A polysaccharide with high uronic acid content from the roots of Panax ginseng was found to inhibit the ability of Porphyromonas gingivalis to agglutinate erythrocytes . This polysaccharide showed a strong inhibitory activity (minimum inhibitory concentration 0.25 mg/mL), but treatment with pectinase resulted in non-inhibitory hydrolyzed products . In contrast, the inhibition by the acidic polysaccharide from the leaves of Artemisia capillaris was negligible . The carbohydrate composition of the two polysaccharides indicated that the anti-adhesive activity may be correlated with glucuronic acid content, one of the components of glycosaminoglycans . Low molecular weight heparin and sucrose octasulfate revealed stronger inhibitory effects on bacterial binding, than the acidic polysaccharide from P . ginseng.

Planta Med, 2004 Jun, 70(6), 572 - 5
Chemical composition and antifungal activity of the essential oil of Thymbra capitata; Salgueiro LR et al.; The composition and the antifungal activity of the essential oil of Thymbra capitata on Candida, Aspergillus and dermatophyte strains were studied . Twenty-two samples of the essential oils from the aerial parts of the plant were obtained by hydrodistillation and analysed by GC and GC-MS . All samples are of the carvacrol type, with a high content of carvacrol (60.0 - 65.8 %) and its biogenetic precursors, gamma-terpinene (8.2 - 9.5 %) and p-cymene (6.0 - 7.5 %) . The minimal inhibitory concentration (MIC) and the minimal lethal concentration (MLC) were used to evaluate the antifungal activity against Candida (7 clinical isolates and 3 ATCC type strains), Aspergillus (5 clinical isolates, 2 CECT and 2 ATCC type strains) and 5 dermatophyte clinical strains . To clarify its mechanism of action on Candida strains, the inhibition of germ tube and a flow cytometry assay with propidium iodide (PI) were used . The oil exhibited antifungal activity for all the tested strains, particularly for dermatophytes, with MIC values ranging from 0.08 to 0.32 microL/mL . Regarding Candida, concentrations lower than the MIC values prevented germ tube formation . After a short incubation time the cells incorporated quickly PI, meaning that the fungicidal effect is mainly due to direct lesion of the membrane.

Planta Med, 2004 Jun, 70(6), 569 - 71
Investigation of the antifungal activity of caledonixanthone E and other xanthones against Aspergillus fumigatus; Larcher G et al.; Among the different xanthones previously isolated from the stem bark of Calophyllum caledonicum, caledonixanthone E presented the strongest activity (MIC (80) = 8 microg/mL) in acidic conditions (pH 3) against the human pathogenic fungus Aspergillus fumigatus . Phase-contrast microscopy studies suggested the assembly or synthesis of cell wall components as the target of the drug . Moreover, the use of fluorescent lectins further supported an impact of caledonixanthone E on the synthesis of chitin, the major structural polysaccharide of the fungal wall . These results suggest that caledonixanthone E may be an interesting model for the design of new antifungal drugs.

Planta Med, 2004 Jun, 70(6), 509 - 14
Antitubercular constituents of Valeriana laxiflora; Gu JQ et al.; Antitubercular bioassay-guided fractionation of the n-hexane- and CH (2)Cl (2)-soluble extracts of the above-ground biomass and roots of Valeriana laxiflora led to the isolation of a new iridolactone, (4R,5R,7S,8S,9S)-7-hydroxy-8-hydroxymethyl-4-methyl-perhydrocyclopenta{ c}pyran-1-one ( 1), and a new lignan, (+)-1-hydroxy-2,6-bis- epi-pinoresinol ( 2), along with eleven known compounds, betulin ( 3), betulinic acid ( 4), 5,7-dihydroxy-3,6,4'-trimethoxyflavone ( 5), 23-hydroxyursolic acid ( 6), oleanolic acid ( 7), tricin ( 8), ursolic acid ( 9), ferulic acid, (+)-1-hydroxypinoresinol, prinsepiol, and 5,7,3'-trihydroxy-4'-methoxyflavone . The structures of compounds 1 and 2 were elucidated on the basis of spectroscopic evidence . The absolute stereochemistry of 1 was determined by chemical transformations and Mosher ester procedures . In a microplate alamar blue assay against Mycobacterium tuberculosis, compounds 2 - 9 exhibited minimum inhibitory concentrations (MIC) of 15.5 - 127 microg/mL, while the other isolates were regarded as inactive (MIC > 128 microg/mL) . In addition, all the isolates were tested for cytotoxicity against African green monkey Vero cells in order to evaluate their selectivity potential.

Bioorg Med Chem Lett, 2004 Aug 2, 14(15), 3979 - 83
Aza-boronic acids as non-beta-lactam inhibitors of AmpC-beta-lactamase; Buzzoni V et al.; With the aim of improving the ability of non-beta-lactam inhibitors to inhibit AmpC-beta-lactamase, a series of 3-aza-phenyl-boronic acid derivatives was obtained using in parallel synthesis . The molecules were tested against Escherichia coli AmpC-beta-lactamase . The best inhibitors, 3-(2-hydroxy-naphthalen-1-ylazo)-phenyl-boronic acid (12) and 3-(2,4-dihydroxy-naphthalen-1-ylazo)-phenyl-boronic acid (14), showed apparent inhibition constant values (K(i)) of 0.3 and 0.45 microM and increased the potency of the semi-synthetic cephalosporin antibiotic, ceftazidime, lowering its minimum inhibitory concentration (MIC) value of 50%, against Gram-negative bacteria strains, producing high levels of AmpC-beta-lactamase.

Chem Biol Interact, 2004 Jun 30, 148(1-2), 1 - 10
Studies on the methyl isocyanate adducts with globin; Mraz J et al.; Isocyanates such as methylisocyanate (MIC), an intermediate in the synthesis of carbamate pesticides, or diisocyanates, used in the production of plastics, are highly reactive toxic compounds that spontaneously bind to biological macromolecules . In vivo formation of stable adducts with blood protein globin offers possibilities for biomonitoring of internal exposure to various reactive species . Thus, biomonitoring of the isocyanates through determination of their specific adducts with globin is a challenge . In this study, we characterized the adducts formed in human globin upon treatment with 100-fold molar excess of MIC . The globin was subject to enzymatic hydrolysis with pronase, and the hydrolysate was analysed by high performance liquid chromatography with positive atmospheric pressure chemical ionization mass spectrometric detection (HPLC/APCI-MS) . The two major MIC adducts were those with N-terminal Val and side-chain of Lys, as confirmed by comparison with the synthetic standards . About 20 other adducts were observed, and several of them were tentatively identified using their MS and MS/MS spectra . Whereas detection of the adducts with Tyr and His was expected, the adducts with Trp and Phe, and a Lys adduct containing two MIC moieties, were probably analytical artifacts resulting from the transcarbamoylation during globin hydrolysis rather than products of direct carbamoylation . The other detected products were MIC-Val-His, derived from the N-terminal dipeptide of globin beta-chain, and dipeptides consisting of MIC-Lys attached to Gly, Val, Leu, Thr, and Glu . Failure to detect the corresponding non-modified dipeptides suggests that the pronase action may be hampered by the amino acid modification . MIC is known as a metabolic intermediate of the industrial solvents N,N-dimethylformamide (DMF) and N-methylformamide (MF) in humans and rats . The HPLC/APCI-MS analysis of globin from rats injected with DMF or MF, 1000 mg/kg, revealed the presence of the MIC adducts with both Val and Lys . The level of the Val adduct in globin from the DMF-dosed rats, determined using Edman degradation and GC/MS, was ca . 40 nmol/g, which is a level common in workers occupationally exposed to DMF . This suggests that also the Lys adduct in such human globin samples can be feasible to analysis and is therefore considered for further studies as a potential biomarker of exposure to DMF.

Pharmacotherapy, 2004 Jun, 24(6), 803 - 7
Meropenem administered as a prolonged infusion to treat serious gram-negative central nervous system infections; Capitano B et al.; The treatment of gram-negative infection of the central nervous system (CNS) presents a clinical challenge due to antibiotic resistance and difficulties with penetration into the cerebrospinal fluid (CSF) . Two patients with gram-negative CNS infections were treated successfully with high-dose, prolonged infusions of meropenem . The CSF meropenem concentrations exceeded the minimum inhibitory concentration of the pathogen for virtually the entire dosing interval in both cases . Our experience demonstrates that dosage modification to maximize pharmacodynamic targets and bactericidal activity may be practically applied to optimize antibiotic treatment for difficult-to-treat CNS infections.

J Colloid Interface Sci, 2004 Aug 1, 276(1), 231 - 8
Synthesis and surface properties of anionic gemini surfactants with amide groups; Yoshimura T et al.; Novel anionic gemini surfactants, 1,2-bis(N-beta-carboxypropanoyl-N-alkylamino)ethane (2CnenAm; n is hydrocarbon chain length of 6, 8, 10, 12, or 14), with two hydrocarbon chains, two carboxylate groups, and two amide groups, were synthesized by three-step reactions . Their solution properties were characterized by equilibrium and dynamic surface tension, steady-state fluorescence spectroscopy of pyrene, and dynamic light-scattering techniques . The surface tension measurements of 2CnenAm give low critical micelle concentrations (cmc), great efficiency in lowering the surface tension, and strong adsorption at air/water interface . Gemini surfactants behave normally with the logarithm of cmc decrease linearly with the chain length . In addition, adsorption and micellization behavior of 2CnenAm was estimated by parameter of pC20, cmc/C20, and standard free energy (DeltaG(0)mic and DeltaG(0)ads); they are significantly influenced by hydrocarbon chain length, and the adsorption is promoted more than the micellization as chain length becomes longer . The results of dynamic light-scattering and fluorescence quenching indicate that small micelles of 2CnenAm are observed at the concentrations above the cmc, and further large particles are also seen . Further, from the dynamic surface tension measurements, it is found that the shorter hydrocarbon chain length of 2CnenAm, the faster the rate of decrease of surface tension.

J Nat Prod, 2004 Jun, 67(6), 968 - 72
New bioactive prenylflavonoids and dibenzocycloheptene derivative from roots of Dendrolobium lanceolatum; Kanokmedhakul S et al.; Two new flavanones (1 and 2), a new flavan (3), and a new rare dibenzocycloheptene derivative (4) together with a known flavan, 4'-hydroxy-2' ',2' 'dimethyl-pyranoflavan (5), were isolated from the roots of Dendrolobium lanceolatum . Their structures were established on the basis of spectral evidence, and an X-ray analysis was performed to confirm the structure of 4 . Compounds 1-3 exhibited antimalarial activity with IC50 values of 2.6, 3.3, and 3.1 microg/mL, respectively . Compounds 1-5 showed moderate antimycobacterial activity with MIC values of 6.3, 12.5, 25, 25, and 50 microg/mL, respectively . In addition, 1 showed strong cytotoxicity against cancer cell lines KB, BC, and NCI-H187 with IC50 values of 1.2, 1.6, and 0.6 microg/mL, respectively, while 2 showed moderate cytotoxicity against the NCI-H187 cell line with an IC50 value of 8.1 microg/ mL.

J Nat Prod, 2004 Jun, 67(6), 938 - 41
Antimycotic spirostanol saponins from Solanum hispidum leaves and their structure-activity relationships; Gonzalez M et al.; A new spirostanol saponin, together with three known saponins, were isolated from the leaves of Solanum hispidum . The structure of the new saponin was elucidated as 6alpha-O-beta-D-quinovopyranosyl-(25S)-5alpha-spirostan-3beta-ol (1) on the basis of spectroscopic analysis (1H NMR, 13C NMR, 1H-1H COSY, HMQC, HMBC, and HRFABMS) . All of the isolated compounds showed antimycotic activity . The most active compound was 6alpha-O-{beta-D-xylopyranosyl-(1-->3)-beta-D-quinovopyranosyl}-(25S)-5alpha-spirostan-3beta-ol (2) (MIC = 25 microg/mL against both Trichophyton mentagrophytes and T . rubrum) . The structure-activity relationships of the isolated compounds and those isolated from S . chrysotrichum are discussed.

Antimicrob Agents Chemother, 2004 Jul, 48(7), 2742 - 6
In vitro susceptibilities of Madurella mycetomatis to itraconazole and amphotericin B assessed by a modified NCCLS method and a viability-based 2,3-Bis(2-methoxy-4-nitro-5- sulfophenyl)-5-{(phenylamino)carbonyl}-2H-tetrazolium hydroxide (XTT) assay; Ahmed AO et al.; Susceptibilities of Madurella mycetomatis against amphotericin B and itraconazole in vitro were determined by protocols based on NCCLS guidelines (visual reading) and a 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-{(phenylamino) carbonyl}-2H-tetrazolium hydroxide (XTT) assay for fungal viability . The XTT assay was reproducible and sensitive for both antifungals . Itraconazole (MIC at which 50% of the isolates tested are inhibited {MIC(50)}) of 0.06 to 0.13 mg/liter) was superior to amphotericin B (MIC(50) of 0.5 to 1.0 mg/liter).

Antimicrob Agents Chemother, 2004 Jul, 48(7), 2524 - 30
Molecular characterization of benzimidazole resistance in Helicobacter pylori; Mills SD et al.; A family of benzimidazole derivatives (BI) was shown to possess potent and selective activity against Helicobacter pylori, although the precise cellular target of the BIs is unknown . Spontaneous H . pylori mutants were isolated as resistant to a representative BI (compound A) . Genomic DNA was isolated from a BI-resistant mutant, transformed into a BI-sensitive strain, and found to be sufficient to confer BI resistance . The resistance determinant was localized to a 17-kb clone after screening a lambda-based genomic library constructed from the BI-resistant strain . Upon sequencing and mapping onto the H . pylori strain J99 genome, the 17-kb clone was shown to contain the entire nuo operon (NADH:ubiquinone oxidoreductase) . Further subcloning and DNA sequencing revealed that a single point mutation in nuoD was responsible for BI resistance . The mutation resulted in a G398S amino acid change at the C terminus of NuoD . Thirty-three additional spontaneous BI-resistant mutants were characterized . Sequencing of nuoD from 32 isolated mutants revealed three classes of missense mutation resulting in amino acid changes in NuoD: G398S, F404S, and V407M . One BI-resistant isolate did not have a mutation in nuoD . Instead, a T27A amino acid change was identified in NuoB . MIC testing of the wild-type H . pylori strain and four classes of nuo mutants revealed that all NuoD mutant classes were hypersensitive to rotenone, a known inhibitor of complex I (NADH:ubiquinone oxidoreductase) suggested to bind to NuoD . Further, a nuoD knockout verified that it is essential in H . pylori and may be the target of the BI compounds.

Antimicrob Agents Chemother, 2004 Jul, 48(7), 2477 - 82
Patterns of amphotericin B killing kinetics against seven Candida species; Canton E et al.; In a previous study tolerance to amphotericin B (AMB) was found among Candida parapsilosis and C . dubliniensis strains by seeding the whole volumes of wells used for MIC determinations, and minimum fungicidal concentrations (MFC) for non-C . albicans Candida strains were demonstrated to be above the levels safely achievable in serum . As an extension of that study, we performed time-kill assays with 26 blood culture isolates (6 C . albicans, 5 C . parapsilosis, 5 C . krusei, 4 C . glabrata, 3 C . lusitaniae, and 3 C . tropicalis isolates), 3 oropharyngeal C . dubliniensis isolates, 3 AMB-susceptible isolates (ATCC 90028, ATCC 22019, ATCC 6254), and 6 AMB-resistant isolates (ATCC 200955, ATCC 200956, ATCC 200950, ATCC 200951, ATCC 200952, ATCC 200953) using RPMI 1640 medium and 0.12 to 32 microg of AMB per ml and determined the numbers of CFU per milliliter at 0, 2, 4, 8, 12, 24, and 48 h . MFCs and time-kill patterns were species specific (MFCs, < or =1 microg/ml for all C . dubliniensis and C . albicans isolates except AMB-resistant strain ATCC 200955; MFCs, 2 to >16 microg/ml for the other isolates) . The times required to reach the fungicidal endpoint (99.9% killing) at four times the MIC were 2 h for C . albicans and C . dubliniensis, 16 h for C . glabrata, 24 h for C . parapsilosis and C . lusitaniae, and > or =40 h for C . tropicalis and C . krusei . The killing rate increased as the AMB concentration was increased up to 2 microg/ml . The highest killing rates were achieved for C . albicans, C . dubliniensis, and C . lusitaniae, while viable C . tropicalis, C . krusei, and C . parapsilosis cells were present after 48 h (MICs, < or =2 microg/ml) when AMB was used at 2 microg/ml . Time-kill curves and MFCs can detect viable cells after 48 h when AMB is used at > or =2 microg/ml . The failure of AMB treatment could be due to its poor killing activity against some species at the concentrations reached in patients' serum.

Antimicrob Agents Chemother, 2004 Jul, 48(7), 2379 - 87
In vitro and in vivo activities of E5700 and ER-119884, two novel orally active squalene synthase inhibitors, against Trypanosoma cruzi; Urbina JA et al.; Chagas' disease is a serious public health problem in Latin America, and no treatment is available for the prevalent chronic stage . Its causative agent, Trypanosoma cruzi, requires specific endogenous sterols for survival, and we have recently demonstrated that squalene synthase (SQS) is a promising target for antiparasitic chemotherapy . E5700 and ER-119884 are quinuclidine-based inhibitors of mammalian SQS that are currently in development as cholesterol- and triglyceride-lowering agents in humans . These compounds were found to be potent noncompetitive or mixed-type inhibitors of T . cruzi SQS with K(i) values in the low nanomolar to subnanomolar range in the absence or presence of 20 microM inorganic pyrophosphate . The antiproliferative 50% inhibitory concentrations of the compounds against extracellular epimastigotes and intracellular amastigotes were ca . 10 nM and 0.4 to 1.6 nM, respectively, with no effects on host cells . When treated with these compounds at the MIC, all of the parasite's sterols disappeared from the parasite cells . In vivo studies indicated that E5700 was able to provide full protection against death and completely arrested the development of parasitemia when given at a concentration of 50 mg/kg of body weight/day for 30 days, while ER-119884 provided only partial protection . This is the first report of an orally active SQS inhibitor that is capable of providing complete protection against fulminant, acute Chagas' disease.

Clin Microbiol Infect, 2004 Jul, 10(7), 662 - 5
In-vitro activity of rifabutin against rifampicin-resistant Mycobacterium tuberculosis isolates with known rpoB mutations; Cavusoglu C et al.; The relationship between resistance to rifampicin and rifabutin and genetic alterations in the rpoB gene of 41 rifampicin-resistant isolates of Mycobacterium tuberculosis was evaluated . Although 35 isolates with rifampicin MICs > or = 32 mg/L were also rifabutin-resistant, six isolates with rifampicin MICs of 2-16 mg/L were susceptible to rifabutin (MIC < or = 0.5 mg/L) . Mutations Asp516Val, Asp516Tyr, Leu533Pro and the double mutation Met515Ile and Leu533Pro influenced susceptibility to rifampicin, but not to rifabutin . All mutations at codons 531 and 526, except one isolate with a His526Cys mutation, correlated with resistance to both compounds.

Pediatr Pulmonol, 2004 Aug, 38(2), 155 - 60
Respiratory morbidity 20 years after RSV infection in infancy; Korppi M et al.; Epidemiological data suggest that respiratory syncytial virus (RSV) infection in early life is a risk factor for later asthma . There are no prospective studies on RSV infection starting from infancy progressing through childhood into adulthood . We followed up a cohort of children, hospitalized for RSV bronchiolitis or RSV pneumonia before age 24 months, until age 18-20 years . The aim of the study was to evaluate early RSV infection as a risk factor for asthma, bronchial reactivity, and lung function abnormalities in young adults . The participants filled in a questionnaire on asthma and asthma-like symptoms . The clinical study included flow-volume spirometry (FVS), methacholine inhalation challenge (MIC), home PEF (peak expiratory flow) monitoring, and skin prick tests (SPT) to common allergens . Asthma was present in 17-22% of 36 index subjects, depending on asthma definition, compared to 11% of 45 controls . Furthermore, FEV% and MEF25 were lower, and MEF50 tended to be lower, in index than in control subjects . One or more abnormal lung function results were found in 16 (44%) index subjects, but only in 5 (11%) controls (P < 0.01) . Bronchial reactivity (PD20 <4,900 microg methacholine) was demonstrated in 16 (46%) index subjects and 14 (32%) controls (NS) . At least one positive SPT result was present in 21 (60%) index subjects; 6 (29%) had asthma (NS vs . nonatopic index subjects); 13 (62%) had abnormal lung function (P < 0.05); and 14 (67%) had bronchial reactivity (P < 0.01) . In the logistic regression adjusted for atopy, as defined by SPT positivity, RSV infection in infancy was an independent risk factor for lung function abnormality (one or more abnormal results in FVS; OR, 5.27; 95% CI, 1.60-17.36), and also for decreased FEV% and MEF50 when these were analyzed separately . However, RSV infection in infancy was not a significant risk factor for asthma or bronchial reactivity . In young adults, lung function abnormalities may be associated with RSV infection which required hospitalization in infancy .

Tuberculosis (Edinb), 2004, 84(3-4), 263 - 74
The use of microarray analysis to determine the gene expression profiles of Mycobacterium tuberculosis in response to anti-bacterial compounds; Waddell SJ et al.; The response of Mycobacterium tuberculosis to six anti-microbial agents was determined by microarray analysis in an attempt to define mechanisms of innate resistance in M . tuberculosis . The gene expression profiles of M . tuberculosis after treatment at the minimal inhibitory concentration (MIC) for 4 h with isoniazid, isoxyl, tetrahydrolipstatin, SRI#221, SR1#967 and SR1#9190 were compared to untreated M . tuberculosis . A common response to drug exposure was defined, and this expression profile overlapped with a number of other mycobacterial stress responses recently identified by microarray analysis . Compound-specific responses were also distinguished including a number of putative transcriptional regulators and translocation-related genes . These genes may contribute to the intrinsic resistance of M . tuberculosis to anti-microbial compounds . Further investigation into these mechanisms may elucidate novel pathways contributing to mycobacterial drug resistance and influence anti-mycobacterial drug development strategies.

Bundesgesundheitsblatt Gesundheitsforschung Gesundheitsschutz, 2004 Apr, 47(4), 352 - 62
{Antibiotic-resistant nosocomial pathogens . Part I: diagnostic and typing methods}; Witte W et al.; For use in human chemotherapy, there are several different substances for nearly each substance group available which cannot all be checked in routine susceptibility testing . If the bacterial resistance mechanisms and cross-resistance conferred by them are known, particular test substances can be selected and the results are interpreted on the basis of cross-resistance . Test substances correspond to those mentioned in guidelines for section sign 23 IfSG (German law on protection against infection) . Due to suboptimal in vitro expression of different resistance mechanisms, it is necessary to perform additional tests besides routine agar-diffusion or microbroth MIC assays . These are preferentially tests for molecular demonstration of resistance genes . Emergence and spread of antibiotic-resistant nosocomial pathogens can be traced by typing . When selecting a typing method, it is important to assess work load, discriminatory power, and reproducibility . In future the availability of microarray technology will enable routine laboratories to demonstrate particular virulence-associated traits.

J Dermatolog Treat, 2004 Apr, 15(2), 104 - 7
Testing of antifungal combinations against yeasts and dermatophytes; Harman S et al.; BACKGROUND: Fungal infections of the nail are a common and chronic problem . The main pathogens responsible for onychomycosis are dermatophytes, yeasts and moulds . Despite significant improvements, approximately 20% of patients with onychomycosis still fail on antifungal therapy . The successful exploitation of drug synergy may provide a useful approach to improve cure rates . METHODS: The minimum inhibitory concentrations (MIC(80)) were recorded for pathogens that are most frequently responsible for onychomycosis against combinations of several antifungal agents, namely, fluconazole, itraconazole, terbinafine and amorolfine . Fractional inhibitory concentrations (FICs) were then calculated from the MIC(80) results and the FIC values for each drug in the combinations added to determine the degree of synergy . A combined value of <1 was taken to suggest synergy; a value of 1-2 indicated an additive effect or indifference; and a combined FIC value of >2 was taken to suggest antagonism . RESULTS: Overall, 46% of amorolfine combinations showed results suggestive of synergy, with the most synergistic results seen against dermatophytes (54%) and moulds (52%) . CONCLUSIONS: Some combinations of drugs may have synergistic activity in vitro; however, the importance of this in a clinical setting is yet to be established, and more studies are justified.

Arch Pharm Res, 2004 May, 27(5), 507 - 11
Antimycobacterial activity and cytotoxicity of flavonoids from the flowers of Chromolaena odorata; Suksamrarn A et al.; From the flowers of Chromolaena odorata (Eupatorium odoratum) four flavanones, isosakuranetin (5,7-dihydroxy-4'-methoxyflavanone) (1), persicogenin (5,3'-dihydroxy-7,4'-dimethoxyflavanone) (2), 5,6,7,4'-tetramethoxyflavanone (3) and 4'-hydroxy-5,6,7-trimethoxyflavanone (4), two chalcones, 2'-hydroxy-4,4',5',6'-tetramethoxychalcone (5) and 4,2'-dihydroxy-4',5',6'-trimethoxychalcone (6), and two flavones, acacetin (5,7-dihydroxy-4'-methoxyflavone) (7) and luteolin (5,7,3',4'-tetrahydroxyflavone) (8) were isolated and identified . Compound 1 exhibited moderate antimycobacterial activity against Mycobacterium tuberculosis with the MIC value of 174.8 microM, whereas compounds 4, 7, and 8 exhibited weak activity with the MIC values of 606.0, 704.2 and 699.3 microM respectively . Compound 7 showed moderate cytotoxicity against human small cell lung cancer (NCI-H187) cells with the MIC value of 24.6 microM, whereas compound 8 exhibited moderate toxicity against NCI-H187 cells and week toxicity against human breast cancer (BC) cells with the MIC values of 19.2 and 38.4 microM respectively.

Arch Pharm Res, 2004 May, 27(5), 502 - 6
Antituberculosis agents X . Synthesis and evaluation of in vitro antituberculosis activity of 2-(5-nitro-2-furyl)- and 2-(1-methyl-5-nitro-1H-imidazol-2-yl)-1,3,4-thiadiazole derivatives; Foroumadi A et al.; Two series of 2-(5-nitro-2-furyl)- and 2-(1-methyl-5-nitro-1H-imidazol-2-yl)-5-propyl, allyl and propargyl)thio-1,3,4-thiadiazoles (6a-f) and 2-(5-nitro-2-furyl)- and 2-(1-methyl-5-nitro-1H-imidazol-2-yl)-5-(nitrobenzyl)thio-1,3,4-thiadiazole derivatives (8a-f) have been synthesized and evaluated against Mycobacterium tuberculosis, as part of the TAACF TB screening program under direction of the US National Institute of Health, the NIAID division . Primary screening was conducted at a single concentration, 6.25 microg mL(-1), against M . tuberculosis H37Rv in BACTEC 12B medium, using the Microplate Alamar Blue Assay (MABA) . The minimum inhibitory concentration (MIC) was determined for the compounds that demonstrated > or = 90% growth inhibition in the primary screening . A varying degree of antituberculosis activity (from 0-97% of growth inhibition) was observed with the alkylthio series (6a-f), and the nitroimidazole derivative with a propylthio group (6b) and the nitrofuran derivative with a propargylthio group (6e), were the most active compounds (MIC=3.13 and 1.56 microg mL(-1), respectively) . Among the nitrobenzylthio derivatives (8a-f), all the ortho, meta and para nitrobenzyl isomers in the nitrofuran series exhibited good antituberculosis activity (MIC=3.13 microg mL(-1)), while the corresponding nitroimidazole analogues were completely inactive (Inhibition=0%).

J Korean Med Sci, 2004 Jun, 19(3), 426 - 30
Molecular analysis of HLA class II-associated susceptibility to neuroinflammatory diseases in Korean children; Oh HH et al.; The work was done to study immunogenetic peculiarities of neuroinflammatory diseases among Korean children . A total of 13 children with neuroinflammatory diseases (8 males and 5 females; mean age 4.6 +/-2.6 yr) were consecutively recruited . Geno-mic typing was performed on their HLA DRB/HLA DQB genes using PCR-SSOP/SSP techniques with gel immunoelectrophoresis . The frequencies of HLA-DR1 *15 in children with acute disseminated encephalomyelitis (ADEM) (31%) and DQB1 *06 in other neuroinflammatory diseases (38%) were significantly increased compared with control subjects . The frequencies of HLA-DRB3 * 0202 (100%), HLA-DRB1 * 1302 (67%), HLA-DRB3 * 0301 (67%), and HLA-DQB1 * 0301 (67%) were significantly increased in children with multiple sclerosis and the frequencies of HLA-DRB1 * 1501 (40%) and HLA-DRB5 * 0101 (40%) were significantly increased in children with ADEM . HLA-DRB1 * 1401, HLA- DRB3 * 0202, and HLA-DQB1 * 0502 were found in children with acute necrotizing encephalopathy . In conclusion, HLA-DR1 * 15 and DQB1 * 06 may be involved in susceptibility to inflammation in Korean children . The frequencies of HLA-DRB1 * 1501, HLA-DRB5 * 0101, HLA-DRB3 * 0301, and HLA-DQB1* 0602 were not as high in Korean children with multiple sclerosis as in western children . However, HLA-DRB3 * 0202 was seen in all children with multiple sclerosis . Our data may provide further evidence that the immunogenetic background of neuroinflammatory diseases in Korean is distinctly different from the ones in western countries . Further studies are necessary to confirm this finding.

J Pediatr Endocrinol Metab, 2004 Apr, 17(4), 679 - 84
Pseudo-precocious puberty caused by a juvenile granulosa cell tumour secreting androstenedione, inhibin and insulin-like growth factor-I; Chan LF et al.; We report a female child who presented at age 3.92 years with a 2-year history of consonant pubertal development caused by a large right-sided ovarian juvenile granulosa cell tumour (JGCT) . Although JGCTs causing pseudo-precocious puberty have been previously described, they remain rare and endocrine data are often incomplete . In this case the tumour was associated with raised serum oestradiol, androstenedione, inhibin and IGF-I . Histological changes were consistent with JGCT . Immunohistochemical studies revealed positive reactivity to MIC-2, inhibin, melan A, IGF-I and IGFBP-2.

Aliment Pharmacol Ther, 2004 Jun 15, 19(12), 1315 - 21
The importance of the level of metronidazole resistance for the success of Helicobacter pylori eradication; Wheeldon TU et al.; AIMS: To evaluate the role of antibiotic susceptibility for the treatment outcome of proton pump inhibitor-dependent and independent Helicobacter pylori eradication regimens . METHODS: In a placebo-controlled clinical study of peptic ulcer patients with H . pylori infection, patients were randomized to receive lansoprazole, clarithromycin and tinidazole twice-daily, clarithromycin and tinidazole once-daily with lansoprazole or with placebo . Helicobacter pylori status was assessed by culture and antibiotic susceptibility by E-test minimal inhibitory concentration (MIC) in 205 clinical isolates . RESULTS: Primary resistance to clarithromycin and metronidazole was 1 and 76%, respectively . In metronidazole susceptible strains eradication rates were similar at > 90% for all treatment groups (P = 0.49) . With low-level metronidazole resistance (4 microg/mL < MIC < 256 microg/mL), eradication rates were similar at >75% (P = 0.80) . The major difference was found at high-level metronidazole resistance (MIC >or= 256 microg/mL) with 95%, 58% and 21% eradication in the lansoprazole, clarithromycin and tinidazole twice-daily, lansoprazole, clarithromycin and tinidazole once-daily and placebo, clarithromycin and tinidazole once-daily groups, respectively (P < 0.001) . CONCLUSION: In the absence of antibiotic resistance, a once-daily therapy of only clarithromycin and tinidazole can achieve a high rate of H . pylori eradication . Such a combination could offer a simpler and cheaper treatment option for developing countries . The standard, twice-daily proton pump inhibitor-based triple therapy was shown to be efficient in H . pylori eradication even in the presence of high-level metronidazole resistance.

J Antimicrob Chemother, 2004 Jul, 54(1), 243 - 6 Epub 2004 Jun 09.
In vitro anti-Helicobacter pylori activity of Extractum liquiritiae, glycyrrhizin and its metabolites; Krausse R et al.; OBJECTIVES: To investigate the in vitro activity of Extractum liquiritiae (EL), glycyrrhizic acid (GL), glycyrrhetinic acid (GA) and a novel lipophilic derivative of glycyrrhetinic acid monoglucuronide (GAMG), acetylated GAMG (aGAMG), against 29 Helicobacter pylori strains . METHODS: The MIC of each compound was determined by the agar dilution method, and the killing kinetics were monitored in brain heart infusion broth ( approximately 10(6)-10(7) cfu/mL) at 0, 4, 24, 48, 72 and 96 h . RESULTS: GA was the most potent compound (MIC(50 /90), 50/100 mg/L), inhibiting 79.3% of the strains at MIC < or =50 mg/L . Clarithromycin-resistant strains were susceptible at 12.5 and 25 mg/L, and metronidazole-resistant strains at 25-50 and at 200 mg/L . The MIC distribution (mg/L) of aGAMG was < or =6.25 (29.2%), 50 (4.2%), 100-200 (12.5%) and > or =400 (54.1%) . EL and GL were less active (MICs >400 mg/L) . GA exhibited rapid, concentration and strain-dependent bactericidal activity . CONCLUSIONS: The potent in vitro activity of GA against H . pylori provides a further explanation for its beneficial effect on peptic ulcers . Its effectiveness against clarithromycin-resistant strains provides hope that it can form the basis for an alternative therapeutic agent against H . pylori.

Clin Ther, 2004 Apr, 26(4), 493 - 501
Pharmacokinetic properties and stability of continuous-infusion meropenem in adults with cystic fibrosis; Kuti JL et al.; BACKGROUND: Meropenem is commonly used to treat lung infections in adults with cystic fibrosis (CF) . Although continuous infusion is the ideal method to maximize the pharmacodynamic properties of this betalactam antibiotic, meropenem is stable for only approximately 4 to 6 hours at room temperature, and its pharmacokinetic (PK) properties, when administered by continuous infusion to patients with CF, are largely unknown . OBJECTIVE: This study was undertaken to determine the PK properties and stability of meropenem when administered to adults with CF by a continuous ambulatory drug-delivery infusion pump stored in a cold pouch between 2 freezer packs . METHODS: This open-label, multidose, randomized, crossover PK study was conducted at the Clinical Research Center at Hartford Hospital (Hartford, Connecticut) . Adults aged > or = 18 years with CF were eligible . Study participants were randomized to receive meropenem 125 mg/h or 250 mg/h (equivalent to 3 g and 6 g, respectively, over 24 hours) by continuous IV infusion for 12 hours . Serum samples were collected throughout the infusion and then for 6 hours after infusion to determine the PK properties (volume of distribution {V(d)}, elimination rate constant, total body clearance {CL}, terminal half-life {t 1/2}, and steady-state concentration {C(ss)}) . Serum meropenem concentrations were assayed using high-performance liquid chromatography, and PK profiles were determined using compartmental analysis . Meropenem stability was ascertained by sampling the drug directly from the infusion pump at prespecified time points . Meropenem tolerability was assessed throughout the study by questioning subjects on how they felt . In addition, laboratory values of serum chemistries and liver enzymes were compared with baseline values . RESULTS: Seven adult volunteers with CF (4 women, 3 men; mean {SD} age, 27 {10} years {range, 19-46 years}) participated in the study . Mean (SD) C(ss) values were 8.31 (0.68) mg/L and 18.50 (3.31) mg/L for the 125-mg/h and 250-mg/h infusion rates, respectively . V(d), CL, and t 1/2 were dose independent and similar between the 2 infusion rates . Meropenem stability was maintained over 12 and 24 hours . Meropenem by continuous infusion was well tolerated . One patient complained of a headache during the study . CONCLUSIONS: In this study of adults with CF, meropenem infusion rates of 125 mg/h and 250 mg/h provided serum drug concentrations greater than the minimum inhibitory concentration for pathogens considered meropenem susceptible (< or =4 microg/mL) and intermediately resistant (8 microg/mL), respectively.

Mycoses, 2004 Jun, 47(5-6), 216 - 21
Susceptibility of sequential Fonsecaea pedrosoi isolates from chromoblastomycosis patients to antifungal agents; Andrade TS et al.; Fourteen Fonsecaea pedrosoi isolates from six chromoblastomycosis patients were submitted to susceptibility testing . Some patients were undergoing treatment with itraconazole (ITZ) and/or cryosurgery with liquid nitrogen . The antifungal agents amphotericin B (AMB), ITZ, fluconazole (FCZ), ketoconazole (KCZ), 5-fluorocytosine (5-FC), and terbinafine (TBF) were tested . AMB and FCZ showed less activity for all isolates . The most active agents were KCZ and TBF . Sequentially isolates from four patients presented ITZ minimal inhibitory concentration (MIC) higher than the previous ones; for two of these patients, response to therapy with this agent was not observed . These results suggest development of microbiologic resistance to ITZ in four instances, two of them coinciding with lack of clinical response to this drug.

Mycoses, 2004 Jun, 47(5-6), 177 - 83
In vitro activities of voriconazole (UK-109, 496), fluconazole, itraconazole and amphotericin B against 132 non-albicans bloodstream yeast isolates (CANARI study); Swinne D et al.; The aim was to evaluate the in vitro activity of voriconazole compared with those of amphotericin B, itraconazole and fluconazole against 132 bloodstream isolates of Candida non-albicans and Saccharomyces cerevisiae species . The minimal inhibitory concentrations (MICs) were determined by an adapted National Committee for Clinical Laboratory Standards (NCCLS) M27-A method using RPMI 1640 as test medium supplemented with 2% glucose . MIC end-points were determined with a spectrophotometer after incubation for 48 h at 35 degrees C . Optical density data were used for the calculation of the MIC end-points . For amphotericin B, the end-point was defined as the minimal antifungal concentration that exerts 90% inhibition compared with the control well growth . For the azoles, the end-points were determined at 50% inhibition of growth . Amphotericin B is highly active with 97% of isolates inhibited by < or =1 microg ml(-1) . Decreased susceptibility or resistance to fluconazole was the rule among C . krusei, which is intrinsically resistant to fluconazole . For C . glabrata isolates, resistance to fluconazole and itraconazole was measured in 13% and 17% of the isolates respectively . Voriconazole was quite active in vitro against all the isolates with a MIC90% of < or =1 microg ml(-1) and we conclude that it may be useful in the treatment of non-albicans bloodstream infections.

Biol Pharm Bull, 2004 Jun, 27(6), 921 - 5
In vitro screening of leishmanicidal activity in myanmar timber extracts; Takahashi M et al.; Seventy-five Myanmar timber extracts belonging to 27 families were examined for their leishmanicidal activities . Some timber extracts had significant leishmanicidal activity, especially extracts of Millettia pendula, which exhibited the most potent activity (MLC 3.1 microg/ml, MIC 1.6 microg/ml) . Other timber extracts showing potent activity included those from Cedrela serrata, Cedrela toona, Cordia fragrantissima, Calophyllum kunstleri, Dalbergia cultrate, Grevillea robusta, Haplophragma adenophyllum, Michelia champaca, and Tectona grandis . From a literature search for reports on the chemical constituents of these plants, most constituents were found to be quinone derivatives or other compounds with unsaturated carbonyl groups.

Biol Pharm Bull, 2004 Jun, 27(6), 806 - 9
Inhibitory effect of miconazole on melanogenesis; Mun YJ et al.; Miconazole (MIC), a regional antifungal agent, has been used worldwide in the treatment of superficial mycosis . However, the effect of MIC on skin pigmentation is not known . In this study, we investigated the inhibitory effect of MIC on melanogenesis in B16 melanoma cells . Tyrosinase activity and melanin content were dose dependently decreased by MIC as compared with untreated cells . The level of tyrosinase protein expression was reduced with treatment MIC . A decrease in cell proliferation was observed in B16 cells treated with MIC 30 microM, indicating that the MIC-induced depigmenting effect was caused by inhibition of melanin synthesis and not by destruction of B16 cells . Furthermore, MIC markedly suppressed alpha-melanocyte stimulating hormone or forskolin-induced tyrosinase activity in B16 cells . Therefore the depigmenting effect of MIC might be due to the inhibition of tyrosinase activity and tyrosinase expression, which eventually slows melanin biosynthesis . These results indicate that MIC may be a useful inhibitor of melanogenesis in B16 cells and suggest that it may have beneficial effects in the treatment of hyperpigmentation disorders such as ephelis and melasma.

Chem Pharm Bull (Tokyo), 2004 Jun, 52(6), 760 - 1
(+)-Bornyl piperate, a new monoterpene ester from Piper aff . pedicellatum roots; Rukachaisirikul T et al.; A new monoterpene ester, (+)-bornyl piperate was isolated from the underground roots of Piper aff . pedicellatum and its structure was elucidated on the basis of spectroscopic evidence and confirmed by X-ray analysis . The compound crystallizes in the triclinic space group P1 with a=7.3232(4) A, b=11.4705(7) A, c=23.2520(14) A, V=1943.6(2) A(3) . This compound showed an antituberculosis activity against Mycobacterium tuberculosis (H(37)Ra strain) with the minimum inhibitor concentration (MIC) of 25 microg/ml.

Appl Environ Microbiol, 2004 Jun, 70(6), 3506 - 11
The weak acid preservative sorbic acid inhibits conidial germination and mycelial growth of Aspergillus niger through intracellular acidification; Plumridge A et al.; The growth of the filamentous fungus Aspergillus niger, a common food spoilage organism, is inhibited by the weak acid preservative sorbic acid (trans-trans-2,4-hexadienoic acid) . Conidia inoculated at 10(5)/ml of medium showed a sorbic acid MIC of 4.5 mM at pH 4.0, whereas the MIC for the amount of mycelia at 24 h developed from the same spore inoculum was threefold lower . The MIC for conidia and, to a lesser extent, mycelia was shown to be dependent on the inoculum size . A . niger is capable of degrading sorbic acid, and this ability has consequences for food preservation strategies . The mechanism of action of sorbic acid was investigated using (31)P nuclear magnetic resonance (NMR) spectroscopy . We show that a rapid decline in cytosolic pH (pH(cyt)) by more than 1 pH unit and a depression of vacuolar pH (pH(vac)) in A . niger occurs in the presence of sorbic acid . The pH gradient over the vacuole completely collapsed as a result of the decline in pH(cyt) . NMR spectra also revealed that sorbic acid (3.0 mM at pH 4.0) caused intracellular ATP pools and levels of sugar-phosphomonoesters and -phosphodiesters of A . niger mycelia to decrease dramatically, and they did not recover . The disruption of pH homeostasis by sorbic acid at concentrations below the MIC could account for the delay in spore germination and retardation of the onset of subsequent mycelial growth.

Eur J Immunogenet, 2004 Jun, 31(3), 105 - 14
Human MHC class I chain related (MIC) genes: their biological function and relevance to disease and transplantation; Collins RW; Major histocompatibility complex (MHC) class I chain related (MIC) molecules show homology with classical human leukocyte antigen (HLA) molecules, but they do not combine with beta2 microglobulin, do not bind peptide and are not expressed on normal circulating lymphocytes . In response to stress, MIC proteins are expressed on the cell surface of freshly isolated gastric epithelium, endothelial cells and fibroblasts and engage the activating natural killer cell receptor NKG2D, which is found on many cells within the immune system . Despite the highly polymorphic nature of MIC genes, only one polymorphic position has been identified that appears to affect the binding of NKG2D . Alleles with a methionine at codon 129 have a 10-50-fold greater capacity to complex NKG2D than alleles with a valine at this position . Renal and pancreatic grafts with evidence of both acute and chronic rejection have been shown to express MIC proteins, and anti-MIC antibodies have been identified in the serum of these patients . Some MIC molecules which are expressed by tumours appear to shed and solubilize in plasma . This soluble form of MIC engages cells expressing NKG2D, rendering them inactive, and impairs tumour cytolysis . Similarly, a protein encoded by human cytomegalovirus (CMV) prevents MICB surface expression and subsequent NKG2D interaction . Whereas the benefit of solid organ transplantation may be hindered by the expression of MIC molecules on grafts, tumours and viruses may take advantage of the expression of MIC molecules on transformed and virus-infected cells in order to evade this recognition pathway.

J Microbiol Methods, 2004 Jul, 58(1), 87 - 100
Methods evaluating vanadium tolerance in bacteria isolated from crude oil contaminated land; Bell JM et al.; Investigations into bacterial responses to vanadium are rare, and in this study were initiated by isolating cultures from crude oil contaminated soil from Russia and Saudi Arabia . Addition of vanadyl sulphate and vanadium pentoxide created acid conditions in the media whilst sodium metavanadate and sodium orthovanadate produced neutral and alkaline effects, respectively . Buffers were introduced for wider comparison of the sample set treatments and to distinguish between the effects of pH and compound toxicity . This study has resulted in the creation of protocols for the pH stabilisation of media containing vanadium compounds and revealed that, although vanadium salts demonstrated some toxic effects, as revealed by MIC and bioluminescence decay tests, the effects were mainly due to pH rather than inherent toxicity of the metal . Capacity for sorption of vanadium to biomass was also investigated .

Phytother Res, 2004 May, 18(5), 414 - 8
In vitro anti-mycobacterial activities of three species of Cola plant extracts (Sterculiaceae); Adeniyi BA et al.; Extracts obtained from three Nigerian Sterculiaceae plants: Cola accuminata, C . nitida and C . milleni were screened for anti-mycobacterium properties using a slow growing Mycobacterium bovis ATCC 35738 (designated BCG Mexican and known to have some virulence in mouse and guinea pig) at 1000 microg/ml using the radiometric (BACTEC) method . The extracts were also tested against six fast growing ATCC strains of M . vaccae using the broth microdilution method.The methanol extracts from both leaves, stem bark and root bark of Cola accuminata and from the leaves and stem bark of C . nitida and C . milleni were not active at the highest concentration of 1000 microg/ml . Only the methanol extract of root bark for both C . nitida and C . milleni were found to be potent against both M . bovis and strains of M . vaccae . The minimum inhibitory concentration (MIC) of C . nitida against M . bovis is 125 microg/ml while the MIC of C . milleni against M . bovis is 62.5 microg/ml after at least 6 days of inhibition with growth index (GI) units lesser than or equal to the change in GI units inoculated with a 1/100 of the BACTEC inoculum for a control vial.The minimum inhibitory concentration of C . milleni against the six ATCC strain of M . vaccae ranged from 62.5 microg/ml to 250 microg/ml while for C . nitida ranged from 500 microg/ml to above 1000microg/ml . Evidently, C . milleni has the highest inhibitory activity against both M . bovis and strains of M . vaccae used . Rifampicin, the positive control used has strong activity against M . bovis at the tested concentration of 5 microg and 10 microg/ml and 4 to 8 microg/ml against the six strains of M . vaccae .

Biosci Biotechnol Biochem, 2004 May, 68(5), 1027 - 34
Anti-microbial action against verotoxigenic Escherichia coli O157:H7 of nitric oxide derived from sodium nitrite; Morita H et al.; The levels of verotoxin-1 and verotoxin-2 released by verotoxigenic Escherichia coli O157:H7 treated in vitro with sodium nitrite, sodium chloride and several antibiotics were evaluated . Of the three strains of E . coli O157:H7 used in this study, two strains produced both verotoxin-1 and verotoxin-2, and one strain produced only verotoxin-2 . Treatment of E . coli O157:H7 with sodium nitrite (6000 mg/l, minimum inhibitory concentration) did not increase the levels of verotoxin-1 and verotoxin-2 compared with a treatment by sodium chloride or antibiotics . When the electron paramagnetic resonance spectrum of sodium nitrite-treated bacterial cells was examined at 77 K to clarify the mechanism for the anti-bacterial activity of nitric oxide derived from sodium nitrite, electron paramagnetic resonance signals with g-values of 2.035 and 2.010 were observed . These were identified as being derived from iron-nitric oxide complexes . It appears that the dinitrosyl iron complexes in the E . coli O157:H7 cells were generated from the reaction of iron-sulfur proteins (enzymes) with nitric oxide formed by the reduction of sodium nitrite . The amount of ATP was decreased by the presence of sodium nitrite in the cell suspension . These findings indicate that nitric oxide derived from sodium nitrite penetrated the cells and inactivated enzymes related to the respiratory chain.

J Nat Prod, 2004 May, 67(5), 851 - 7
Enniatins of Fusarium sp . strain F31 and their inhibition of Botrytis cinerea spore germination; Pohanka A et al.; A spectrum of enniatins was isolated from Fusarium sp . strain F31 by bioassay-guided isolation directed against Botrytis cinerea . Two new enniatins, J(2) (7) and J(3) (8), were co-isolated and both contained, in addition to three hydroxyisovaleric acid units, N-methylated l-alanine, l-valine, and l-isoleucine units, differing only in their primary sequence . Two other enniatins, named enniatin J(1) (1) and enniatin K(1) (6), each containing two N-Me-l-Val units and one N-Me-l-Ala or alpha-N-Me-l-butyric acid unit, respectively, were isolated for the first time without directed biosynthesis . The enniatin structures were elucidated by spectroscopic and chemical methods, and the absolute configuration of the amino acids (l) and hydroxyisovaleric acid (d) was consistent with all previously isolated enniatins . The known enniatins B (2), B(1) (4), B(2) (5), and B(4) (3) were also isolated . The minimum inhibitory concentration of pure enniatins against Botrytis cinerea was 75 microg/mL.

Int J Antimicrob Agents, 2004 Mar, 23(3), 307 - 10
Evaluation of a triple-drug combination for treatment of experimental multidrug-resistant pneumococcal meningitis; Lee H et al.; To evaluate the therapeutic efficacy of ceftriaxone + vancomycin + rifampicin (CVR) in the treatment of pneumococcal meningitis caused by a multidrug-resistant strain, single-drug regimens (ceftriaxone 100 mg/kg, rifampicin 15 mg/kg, or vancomycin 20 mg/kg), double-drug regimens (ceftriaxone + vancomycin {CV} and ceftriaxone + rifampicin {CR}) and a triple-drug combination (CVR) with or without dexamethasone were compared in a rabbit meningitis model . Meningitis was induced by a highly penicillin-resistant (MIC 2 mg/l) and ceftriaxone-resistant (MIC 4 mg/l) pneumococcal strain . Final therapeutic efficacy was evaluated by the bacterial concentration at 24 h, and the bacterial killing rate was also evaluated . All combination regimens were superior to ceftriaxone or vancomycin single-drug regimens with regard to sterilisation of CSF and bacterial killing rate . Rifampicin was as effective as combination regimens . Regardless of dexamethasone, therapeutic efficacy of CVR and CR were superior to that of CV . CVR showed comparable therapeutic efficacy to CR . Data suggested that CVR would not have additional therapeutic benefit over CR during the initial 24 h of treatment.

Int Urol Nephrol, 2003, 35(3), 311 - 2
Primary primitive neuroectodermal tumor (PNET) of the kidney with venous thrombus; Sivaramakrishna B et al.; Primitive neuroectodermal tumors of the kidney are rare, the diagnosis usually being made at histopathology . A young adult presented with a painful left renal mass . CT Scan of the abdomen revealed a large necrotic tumor of the left kidney . At surgery the patient was found to have a venous thrombus confined to the renal vein . Radical nephrectomy was done . Histopathology showed a round cell neoplasm with typical Homer Wright rosette formation and positive staining for neuron specific enolase (NSE) and MIC-2 on immunohistochemistry . The patient is undergoing multidrug chemotherapy and is alive and well at a follow up of nine months.

J Infect Chemother, 2004 Apr, 10(2), 128 - 30
In vitro exposure to macrolide antibiotics in Helicobacter pylori strains isolated from children; Fujimura S et al.; Clarithromycin resistance of Helicobacter pylori is a serious problem in eradication therapy . We investigated whether the use of maclorides (clarithromycin, erythromycin, and azithromycin) induces clarithromycin resistance in the organism . Twenty H . pylori strains were isolated from pediatric patients with gastrointestinal symptoms . The minimum inhibitory concentrations (MICs) of each macrolide antibiotic were determined by the Etest . Among these, 17 strains susceptible to macrolide antibiotics were used for the in vitro induction of drug resistance . In each of these 17 strains of H . pylori, 30-day exposure to clarithromycin in experiments for in vitro induction did not change the MIC of any antibiotic, nor did it induce either the A2143G or the A2144G mutation in the 23S rRNA gene . These results suggest that the use of macrolide antibiotics does not induce clarithromycin resistance in H . pylori by 23S rRNA gene mutation.

Shanghai Kou Qiang Yi Xue, 1992 Jun, 1(1), 22 - 4
{A study on resorbable local metronidazol delivery devices}; Fan MW et al.; A local metronidazol (MTZ) delivery using resorbable base material was studied . In vitro MTZ was almost completely released within 24 hrs from the hydroxypropylmethyl cellullouse(HPMC) strips.In vivo,release of the drug strips was also measured in six patients who had deep pockets of more than 5mm.MTZ-containing HPMC strip was inserted in the pocket.The average amount of MTZ in the gingival crevicalar fluid(GCF) 24 hrs after insertion of strip was 40microg/ml.(much higher than the reported MIC of some microorganisms associated with adult periodontitis).In vitro there were significant inhibitory effects of MTZ containing strips on the microorganisms associated with periodontal disease.The prevalence of spirochetes and motil rods,and gingival index(GI) was remarkably reduced in the MTZ administered pockets,It is suggest that this new direct drug delivery system with HPMC used as base material could be used to treat periodontal disease.

Antimicrob Agents Chemother, 2004 Jun, 48(6), 2081 - 4
In vitro activities of garenoxacin and levofloxacin against Chlamydia pneumoniae are not affected by presence of Mycoplasma DNA; Smith RP et al.; We studied 20 Chlamydia pneumoniae isolates obtained from respiratory sites and atheroma tissue of patients from various geographic areas to determine the susceptibilities of these isolates to a new des-fluoroquinolone, garenoxacin, and to levofloxacin . In addition, we assessed the cultures with these isolates by PCR for the presence or absence of Mycoplasma sp . DNA . Both the MIC at which 90% of isolates are inhibited (MIC(90)) and the minimal bactericidal concentration at which 90% of isolates are killed (MBC(90)) for garenoxacin were 0.06 microg/ml, and both the MIC(90) and the MBC(90) for levofloxacin were 2.0 microg/ml . The activity of garenoxacin against C . pneumoniae was 32-fold greater than that of levofloxacin . Mycoplasma sp . DNA was detected by PCR in 17 of 20 cultures . Mycoplasma amplicons from five Mycoplasma DNA-positive C . pneumoniae cultures were sequenced and found to represent four Mycoplasma species . Our data demonstrate that C . pneumoniae cultures frequently contain Mycoplasma DNA and that its presence in C . pneumoniae cultures does not appear to affect the susceptibility results for the two fluoroquinolones that we tested.

Antimicrob Agents Chemother, 2004 Jun, 48(6), 2007 - 13
In vitro interactions between amphotericin B, itraconazole, and flucytosine against 21 clinical Aspergillus isolates determined by two drug interaction models; Te Dorsthorst DT et al.; Combination therapy of flucytosine (5FC) with other antifungal agents could be of use for the treatment of invasive aspergillosis . However, interpretation of the results of in vitro interactions is problematic . The fractional inhibitory concentration (FIC) index is the most commonly used method, but it has several major drawbacks in characterizing antifungal drug interaction . Alternatively, a response surface approach using the concentration-effect relationship over the whole concentration range instead of just the MIC can be used . We determined the in vitro interactions between amphotericin B (AMB), itraconazole, and 5FC against 21 Aspergillus isolates with a broth microdilution checkerboard method that employs the dye MTT {3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide} . FIC indices based on three different MIC endpoints (MIC-0, MIC-1, and MIC-2) and the interaction coefficient alpha were determined, the latter by estimation from the response surface approach described by Greco et al . (W . R . Greco, G . Bravo, and J . C . Parsons, Pharmacol . Rev . 47:331-385, 1995) . The value obtained for the FIC index was found to be dependent on the MIC endpoint used and could be either synergistic, indifferent, or antagonistic . The response surface approach gave more consistent results . Of the three combinations tested, the AMB-5FC combination was the most potent in vitro against Aspergillus spp . We conclude that the use of the response surface approach for the interpretation of in vitro interaction studies of antifungals may be helpful in order to predict the nature and intensity of the drug interaction . However, the correlation of these results with clinical outcome remains difficult and needs to be further investigated.

Antimicrob Agents Chemother, 2004 Jun, 48(6), 1941 - 7
Determination of the in vivo pharmacodynamic profile of cefepime against extended-spectrum-beta-lactamase-producing Escherichia coli at various inocula; Maglio D et al.; Cefepime was evaluated in vivo against two inoculum sizes of four strains of Escherichia coli that produced extended-spectrum beta-lactamases (ESBLs) in a murine neutropenic thigh infection model to characterize the pharmacodynamic activity of cefepime in the presence of ESBL-producing bacteria and to evaluate if differences in lengths of cefepime exposure are required with various inocula . Three strains possessed a single enzyme each: TEM-10, TEM-12, and TEM-26 . The fourth strain possessed two TEM-derived ESBLs and a third uncharacterized enzyme . Two non-ESBL-producing E . coli strains were included for comparison . Mice received various doses of cefepime to achieve a spectrum of percentages of time the drug was above the MIC (%T>MICs) for each isolate at both inocula . No significant difference in cefepime exposure was required to achieve similar bactericidal effects for ESBL- and non-ESBL-producing isolates when the starting inoculum was 10(5) CFU of E . coli per thigh . The increased MICs observed in vitro for the ESBL-producing strains at 10(7) CFU/ml did not predict the amount of exposure required to achieve a comparable level of bactericidal activity in vivo at the corresponding starting inoculum of 10(7) CFU/thigh . Compared to the cefepime exposure in tests with the lower inoculum (10(5) CFU/thigh), less exposure was required when the starting inoculum was 10(7) CFU/thigh (%T>MIC, 6% versus 26%), such that similar doses (in milligrams per kilogram of body weight) produced similar bactericidal effects with both inocula of ESBL-producing isolates . Equivalent exposures of cefepime produced similar effects against the microorganisms regardless of the presence of ESBL production . Pharmacodynamic profiling undertaken with conventional cefepime MIC determinations predicted in vivo microbial outcomes at both inoculum sizes for the ESBL-producing isolates evaluated in this study . These data support the use of conventional MIC determinations in the pharmacodynamic assessment of cefepime.

Br J Cancer, 2004 May 4, 90(9), 1809 - 13
The increase of MICA gene A9 allele associated with gastric cancer and less schirrous change; Lo SS et al.; Since surgical resection is the principal treatment of gastric cancer, early detection is the only effective strategy against this disease at present . Recently, a new polymorphic gene family, the major histocompatibility complex class I chain-related (MIC) genes located about 40 kb centromeric to HLA-B gene has been proposed . This family consists of five genes (A, B, C, D and E) . Among them, MICA has five various alleles (A4, A5, A5.1, A6 and A9), which can be used as a polymorphic marker for genetic mapping and for disease susceptibility . The MICA polymorphism was studied in our gastric cancer patients to see if there is any possible correlation with genetic predisposition and clinicopathological factors . Genomic DNA was extracted from fresh or frozen peripheral blood leukocytes in 107 patients with gastric adenocarcinoma who underwent gastrectomy in our hospital and 351 noncancer controls . MICA polymorphism was analysed by using PCR-based technique . The results showed both phenotypic and allele frequencies of allele A9 in patients with gastric cancer were significantly higher than controls (33 vs 17.6%, P=0.005; 17 vs 9.9%, P=0.02) . Gastric adenocarcinoma with allele A9 was associated with less schirrous change than those without (P=0.014) . MICA gene A9 allele might confer the risk of gastric cancer and associate with less schirrous change . The mechanisms among them deserve further investigation.

J Antimicrob Chemother, 2004 Jul, 54(1), 240 - 2 Epub 2004 May 18.
Nitroimidazole resistance genes (nimB) in anaerobic Gram-positive cocci (previously Peptostreptococcus spp.); Theron MM et al.; OBJECTIVES: To investigate metronidazole resistance and the prevalence of nitroimidazole (nim) genes in clinically isolated anaerobic Gram-positive cocci . METHODS: Metronidazole susceptibility was determined in 99 strains of anaerobic Gram-positive cocci and PCR amplification for the nim gene carried out on 61 strains (metronidazole MIC > or =0.5 mg/L) . RESULTS: The nimB gene was detected in 34% (21/61) of the strains . These included two highly resistant Finegoldia magna strains (MICs >128 mg/L) . The nimB gene was, however, also demonstrated in 90% (19/21) of susceptible strains . CONCLUSIONS: Although the nimB gene may be implicated in the high-level metronidazole resistance in 2 F . magna strains, the alarmingly high prevalence of the nimB gene in anaerobic Gram-positive cocci cannot be directly associated with resistance and the possibility of a silent nimB gene should be considered.

Res Microbiol, 2004 May, 155(4), 275 - 82
Pleiotropic effect of sodium arsenite on Escherichia coli; Gualco L et al.; The effects of sodium arsenite at a sub-MIC concentration (25 mg/l) upon different bacterial functions were studied . This compound reduced the killing activity of nalidixic acid, amikacin, and meropenem . It also promoted the loss of F'lac from bacterial hosts and increased the number of recombinants in conjugation and transduction experiments . Transposition of Tn9 was also enhanced by the salt . In addition, sodium arsenite abolished the lethal effect of temperature on thermosusceptible DNA synthesis mutants in a similar manner to that seen in an anaerobic environment . Finally at a low dose, it induced the SOS response, and the related production of recA-dependent enzymes was reduced as the sodium arsenite concentration increased . It has been suggested that arsenite primarily affects the uvrA gene product, influencing the other bacterial functions studied . The energetic depletion caused by this compound appears to play a role in the activity of autolytic enzymes.

Curr Infect Dis Rep, 2004 Jun, 6(3), 172 - 180
Prevention and Treatment of Postsurgical Head and Neck Infections; Fraioli R et al.; Postoperative wound infection is an important cause of postsurgical morbidity . Efforts to reduce the incidence of wound infection are enhanced through appropriate preoperative preparation, adherence to excellent surgical technique, and the provision of outstanding postoperative care . Many head and neck surgical procedures are undertaken in an environment contaminated by saliva containing a large inoculum of potentially pathogenic bacteria . Prospective randomized clinical trials have demonstrated that perioperative antibiotic prophylaxis reduces the risk for postoperative wound infection . Effective antibiotic prophylaxis requires that the antibiotic be effective against normal oral flora . Antibiotics should be administered before wound contamination . The dose of the antibiotic should exceed the minimal inhibitory concentration needed for the normal flora . The antibiotic administration can cease within 24 hours of surgery.

J Pharm Pharmacol, 2004 May, 56(5), 663 - 9
Trypanocidal and antifungal activities of p-hydroxyacetophenone derivatives from Calea uniflora (Heliantheae, Asteraceae); do Nascimento AM et al.; The dichloromethane extract of underground parts of Calea uniflora (Heliantheae, Asteraceae) exhibited trypanocidal and antifungal activities . Four p-hydroxyacetophenone derivatives were isolated as the main compounds: 2-senecioyl-4-(hydroxyethyl)-phenol (1), 2-senecioyl-4-(angeloyloxy-ethyl)-phenol (2), and two new derivatives, 2-senecioyl-4-(methoxyethyl)-phenol (3) and 2-senecioyl-4-(pentadecanoyloxyethyl)-phenol (4) . 1 and 4 were active towards Trypanosoma cruzi trypomastigotes, reducing their number by 70 and 71% at 500 microg x mL(-1), whereas 2 and 3 were inactive . All the compounds tested showed antifungal activity with minimal inhibitory concentration values between 500 and 1000 microg x mL(-1) against pathogenic Candida spp . and dermatophytes . The isolation, structure elucidation, NMR spectral assignments and bioactivity results of these compounds are reported.

FEMS Microbiol Lett, 2004 May 15, 234(2), 343 - 8
The effect of NAG-thiazoline on morphology and surface hydrophobicity of Escherichia coli; Reid CW et al.; The beta-hexosaminidase inhibitor and structural analog of the putative oxazolium reaction intermediate of lytic transglycosylases, N-acetylglucosamine thiazoline (NAG-thiazoline), was synthesized in 46% overall yield and tested as an inhibitor of Escherichia coli growth . NAG-thiazoline, at concentrations up to 1 mg/ml, was not found to affect the viability of E . coli DH5alpha . However, the compound did induce morphological changes to the cells . Growth of cells in the presence of NAG-thiazoline caused an apparent inhibition of the biosynthesis of the cylindrical regions of the cells such that they became much shorter in length . The surface of these shorter cells was found to be much less hydrophobic compared to untreated cells as determined by the bacterial adhesion to hydrocarbon (BATH) assay . In addition, the co-administration of NAG-thiazoline with 1.7 x MIC concentrations of ampicillin prevented cell lysis suggesting that the compound inhibited autolytic enzymes, in particular the lytic transglycosylases .

Cas Lek Cesk, 2004, 143(3), 159 - 63
{Genetic risk factors in autoimmune diabetes mellitus, their significance and function}; Novota P et al.; Autoimmune diabetes mellitus is characterized by selective destruction of beta pancreatic cells and by cellular infiltration with T- (particularly Th1) and B-lymphocytes . The marker of autoimmunity is the presence of autoantibodies (ICA, IAA, GADab, IA2ab) . Etiology of the autoimmune process is still unknown . It is suggested that the pathogenesis is activated by genetic and environmental factors . Individual predisposition can influence also the onset and progression of the disease . The most important genetic risk factors of autoimmune diabetes mellitus are the HLA class II alleles (DQB1*0302, 0201; DRB1*0301, 0401; DQA1*0301, 0501) and the risk alleles of INS-VNTR of the promoter region . Recent studies have shown various genetic risk factors for the autoimmune diabetes mellitus . Individual predispositions belong to the genetic polymorphisms in cytokine genes (IL-10, IL-12, IL-18) and the microsatellite polymorphism of MHC class I chain-related gene A (MIC-A).

Int J Pharm, 2004 Mar 1, 271(1-2), 41 - 52
Improvement of stability and absorbability of dry insulin powder for inhalation by powder-combination technique; Todo H et al.; The effect of pulmonary absorption enhancers on the stability of active ingredients is an important factor for successful inhalation therapy as well as the effect on pharmacological activity and safety . We examined the effect of pulmonary absorption enhancers on the stability of insulin in dry powders prepared by a spray-drying technique . Although the hypoglycemic effect was greatly improved when a dry insulin powder containing citric acid (MIC SD) was administered, insulin in the MIC SD was unstable compared with the other powders examined . Bacitracin and Span 85, which are potent pulmonary absorption enhancers of insulin formulated in solutions, showed no deteriorative effect on the stability of dry insulin powder . However, they did not improve the hypoglycemic effect of insulin in dry powders . We modified the insulin dosage form with citric acid to improve the insulin stability at room temperature without loss of hypoglycemic activity . MIC Mix was formulated as a combination of insulin powder (MI') and citric acid powder (MC) . MIC Mix showed hypoglycemic activity comparable to MIC SD while the insulin stability was much better than that of MIC SD at a 60 degrees C/dry condition . However, moisture lowered the insulin stability and changed the particle morphology of MIC Mix with time at a 60 degrees C/75% relative humidity condition, suggesting that a package preventing moisture absorption was necessary for the MIC Mix powder.

Med Mycol, 2004 Apr, 42(2), 159 - 63
Antifungal drug response in an in vitro model of dermatophyte nail infection; Osborne CS et al.; Despite terbinafine being fungicidal against Trichophyton rubrum in standard NCCLS assays and rapidly accumulating in nails in vivo, onychomycosis patients require prolonged terbinafine treatment to be cured . To investigate this, we developed a more clinically relevant onychomycosis in vitro test model . Human nail powder inoculated with T . rubrum and incubated in liquid RPMI 1640 salt medium, which did not support growth alone, developed extensive and invasive mycelial growth . Antifungal drugs were added at different concentrations and cultures incubated for 1 to 4 weeks . Fungal survival was determined by spreading cultures on PDA plates without drug and measuring CFU after 1 to 4 weeks incubation . Drug activity was expressed as the nail minimum fungicidal concentration (Nail-MFC) required for 99.9% elimination of viable fungus . Terbinafine Nail-MFC was 4 microg/ml after 1 week exposure, decreasing to 1 microg/ml after 4 weeks exposure, much higher than MFCs < or = 0.03 microg/ml determined in standard NCCLS MIC assays . In contrast, other clinically used drugs were unable to kill T . rubrum after 4 weeks incubation in this model . Invasive mycelial growth on nail appears to protect T . rubrum from the cidal action of systemic drugs, thus providing a rationale for the long treatment periods in onychomycosis.

Ann Hematol, 2004, 83 Suppl 1, S42 - 4
Update on the treatment of cerebral aspergillosis; Schwartz S et al.; In aspergillus infections of the central nervous system (CNS) the mortality risk usually exceeds 90% . Data from case-reports and a recent retrospective study suggest that neurosurgical interventions, such as abscess resections, stereotactic drainages, or the use of intraventricular catheters, might improve the outcome in CNS aspergillosis . However, there is a lack of clear evidence supporting an extensive neurosurgical management in these patients . A major reason for the devastating prognosis in CNS aspergillosis is a poor penetration of antifungal drugs into the CNS, with the exception of voriconazole . Treatment with voriconazole results in measurable drug levels in the cerebrospinal fluid, which may exceed the minimal inhibitory concentration for aspergillus . Moreover, voriconazole brain tissue levels exceed those measured for other antifungal drugs . In a recent retrospective study, a complete or partial response occurred in 35% of patients who were treated with voriconazole for CNS aspergillosis with a survival rate of 31% . These data support the use of voriconazole in this clinical setting . An intense neurosurgical management and higher doses of voriconazole might further improve the outcome in CNS aspergillosis, but this needs to be evaluated in future studies.

Epilepsy Behav, 2004 Apr, 5(2), 224 - 30
Quality of life improvement with conversion to lamotrigine monotherapy; Cramer JA et al.; This report describes the effect on patient-reported quality of life (QOL) after reduction from two drugs to monotherapy with lamotrigine . Patients taking lamotrigine (LTG) with an enzyme-inducing drug were converted to LTG monotherapy for a 12-week follow-up . Changes in QOLIE-31 between baseline and follow-up were compared with physicians' global change ratings and patient-reported health status . Total QOLIE-31 scores increased 10.7 points for patients rated by physicians as having mild improvement, and 17 points for those reported as having moderate to marked improvement . Subscale scores also increased by minimum important change (MIC) amounts (> or = 11.76), with the largest change in Cognition, Energy, Medication Effects, and Seizure Worry subscales . The data also support > or = 11 MIC as a clinically important change in total score for the QOLIE-31 . Exploratory analyses also provide information about MIC for individual subscales (8-18 for physician rated global change, 10-26 for patient-rated global health status change) . This study demonstrates the value of reduction to monotherapy from the patients' and physicians' perspectives.

Nippon Ishinkin Gakkai Zasshi, 2004, 45(2), 83 - 91
{Status and issues of preclinical evaluation of the therapeutic effects of newly developed antifungal agents}; Uchida K et al.; Preclinical evaluation of the efficacy of an antifungal agent is basically conducted by measuring both the in vitro and the in vivo antifungal activity of the drug using appropriate infection models . Although the first requisite for a method measuring in vitro activity is to obtain results with good reproducibility, an additional requirement is that there be good correlation with the in vivo activity, as described later . For the first condition, in recent years the National Committee for Clinical Laboratory Standards in the United States and the Standardization Committee of the Japanese Society for Medical Mycology have proposed reference techniques with the objective of standardizing drug susceptibility testing; these have been used extensively in measuring antifungal activities of novel agents . However, there are several issues involved when these methods are applied to newly developed drugs . First, standard methods are for particular currently available antifungal agents, but MIC determining standards have not been established for other agents . Reproducibility is therefore not guaranteed . Second, there is a question of whether reliable results can be obtained to test an antifungal spectrum with a limited number of fungal species . On the other hand, in vivo evaluation of novel antifungal agents is extremely important to predict the clinical outcome at the preclinical stage . The important requirements for this in vivo experimental system are: use of an animal model of mycosis that resembles the pathophysiology in humans; use of an administration schedule corresponding to that used in clinical studies, and evaluation of the therapeutic effect considering the dose and administration period . This review presents the present status of preclinical evaluation test methods and discusses the issues.

Southeast Asian J Trop Med Public Health, 2003 Sep, 34(3), 627 - 9
Stability of meropenem in normal saline solution after storage at room temperature; Jaruratanasirikul S et al.; The bactericidal activity of meropenem is determined by the time that concentrations in tissue and serum are above the MIC for the pathogens during the dosing interval . Thus, the most effective mode of administering of meropenem is continuous infusion . However, the stability of meropenem reconstituted in solution is influenced by the storage temperature . Until now we have had no data to evaluate the stability of this drug during continuous infusion in a tropical country . The objective of this study was to provide such data . Meropenem 0.5 g and 100 ml normal saline solution were mixed together and stored at room temperature for 8 hours . Half of the solution was stored in a room with air conditioning at 20 degrees C and the other half of the solution was stored in a room without air conditioning at 32 degrees-37 degrees C . The concentrations of meropenem in the solution were measured at 0, 1, 2, 3, 4, 6 and 8 hours after the drug was reconstituted . Twelve lots of (0.5 g meropenem in normal saline) solution were evaluated in each temperature condition . The mean meropenem concentrations reconstituted in normal saline solution decreased 1.66%, 3.31% and 5.80% after 2, 4 and 8 hours storage at 20 degrees C, respectively . Drug concentrations decreased 3.14%, 5.86% and 11.85% after 2, 4 and 8 hours storage at 32 degrees-37 degrees C, respectively . Therefore, we conclude that this agent should not be administered by 8-hour continuous infusion at room temperature in a tropical country.

No To Shinkei, 2004 Mar, 56(3), 237 - 41
{A case of peripheral-type primitive neuroectodermal tumor arising in the dura mater at the frontal base}; Utsunomiya A et al.; A 7-year-old boy was admitted to our hospital because of headache and frequent vomiting . The patient was noted to have papilloedema and mild palsy of the right abducent nerve . Magnetic resonance image (MRI) revealed a large tumor in the frontal base with tumoral hemorrhage . Angiography showed the tumor was fed by anterior meningeal arteries . At surgery, the tumor was arising in the dura mater at the frontal base, and was removed totally . Histological examination showed the tumor to be composed of small cells with uniform round nuclei and minimal cytoplasm . Immunohistochemical studies were positive for MIC-2, NSE, C-KIT, vimentin, Class III-beta tublin and glycogen, but negative for NFP, synaptophysin, chromogranin A and GFAP . MIB-1 labeling index was 40-50% . The tumor was histologically confirmed to be peripheral-type primitive neuroectodermal tumor(pPNET) . Following surgery, he underwent whole brain, whole spine and local radiation therapy(30 Gy in total respectively) and received two 5-day cycles of chemotherapy, consisting of intravenous administration of cisplatin 20 mg/m2/day, etoposide 60 mg/m2/day and IFOS 900 mg/m2/day . After these therapies, follow-up radiological examination showed there was no recurrence of the tumor for 24 months . Intracranial pPNET is rare . Ewing sarcoma and pPNET(ES/pPNET) is the designation given to a family of small round cell tumor arising in bone or soft tissues . Intracranial PNETs are devided into central nervous system PNET(cPNET) and pPNET . It is necessary that intracranial PNETs are divided into two types of PNETs because of different prognosis between these tumors . MIC-2 is a specific marker for pPNET/ES family and is useful in the differential diagnosis of these two types of tumors.

Bioorg Med Chem, 2004 May 15, 12(10), 2501 - 8
Ring-substituted quinolines as potential anti-tuberculosis agents; Vangapandu S et al.; We report in vitro antimycobacterial properties of ring-substituted quinolines (series 1-4) constituting 56 analogues against drug-sensitive and drug-resistant M . tuberculosis H37Rv strains . The most effective compounds 2h (R1 = R2 = c-C6H11, R3 = NO2, series 1) and 13g (R1 = OC7H15, R2 = NO2, series 4) have exhibited an MIC value of 1 microg/mL against drug-sensitive M . tuberculosis H37Rv strain that is comparable to first line anti-tuberculosis drug, isoniazid . Selected analogues (2d, 2g, 2h, 4e, 6b, 13b, 13g, and 14e, MIC: < or = 6.25 microg/mL) upon further evaluation against single-drug-resistant (SDR) strains of M . tuberculosis H37Rv have produced potent efficacy in the range between 6.25 and 50 microg/mL.

FEMS Microbiol Lett, 2004 May 1, 234(1), 99 - 103
Exploring the effectiveness of tazobactam against ceftazidime resistant Escherichia coli: insights from the comparison between susceptibility testing and beta-lactamase inhibition; Bethel CR et al.; Thirteen clinical isolates of Escherichia coli resistant to ceftazidime that possessed an AmpC and other (beta-lactamases were identified . The effectiveness of different formulations of piperacillin/tazobactam to other beta-lactams was compared . Antibiotic susceptibility testing, polymerase chain reaction, amplification of blaTEM, blaSHV and blaAmpC, and enzyme-linked immunosorbent assays to identify AmpC beta-lactamases were performed . Hydrolysis rates were obtained and residual enzymatic activity was determined . Cefepime and ertapenem were more active than piperacillin/tazobactam . In contrast, increasing the relative proportion of tazobactam improved susceptibility testing . Twenty micromolar tazobactam inhibited total beta-lactamase activity (as measured by nitrocefin hydrolysis rates) by greater than 75% against all isolates tested: in 11 of 13 E . coli isolates, total beta-lactamase activity was inhibited by 90% . The observed differences between MIC determinations and susceptibility to enzymatic inactivation by tazobactam against E . coli containing AmpC and other -lactamases may be due to the final tazobactam concentration achieved in the periplasmic space . Factors determining this are critical considerations in assessing beta-lactamase inhibitor potency.

Antimicrob Agents Chemother, 2004 May, 48(5), 1885 - 6
In vitro activity of a novel diaminopyrimidine compound, iclaprim, against Chlamydia trachomatis and C . pneumoniae; Kohlhoff SA et al.; The in vitro activities of iclaprim, a novel dihydrofolate reductase inhibitor, azithromycin, and levofloxacin were tested against 10 strains of Chlamydia trachomatis and 10 isolates of Chlamydia pneumoniae . For C . trachomatis and C . pneumoniae, the iclaprim MIC and minimal bactericidal concentration at which 90% of isolates were inhibited (MIC(90) and MBC(90)) were 0.5 micro g/ml, compared to an azithromycin MIC(90) and MBC(90) of 0.125 micro g/ml and levofloxacin MIC(90)s and MBC(90)s of 1 micro g/ml for C . trachomatis and 0.5 micro g/ml for C . pneumoniae.

Antimicrob Agents Chemother, 2004 May, 48(5), 1699 - 707
Mutant selection window in levofloxacin and moxifloxacin treatments of experimental pneumococcal pneumonia in a rabbit model of human therapy; Croisier D et al.; For some pneumococci the fluoroquinolone MICs are low but the mutant prevention concentrations (MPCs) are high; this difference defines in vitro the mutant selection window (MSW) . We investigated in vivo the bacterial reduction and the occurrence of resistant mutants with moxifloxacin (MFX; 400 mg once daily) or levofloxacin (LVX; 500 mg twice daily) in treatments similar to those in humans with experimental pneumonia due to pneumococci (expPP) exhibiting various MICs and MPCs . The MIC/MPC for MFX and LVX and genotypes were as follows: strain 16089, 0.125/0.125 and 0.5/0.5 (wild type); strain MS1A, 0.25/0.25 and 1/2 (efflux); strain MS2A, 0.25/4 and 1.75/28 (parC79); strain MR3B4, 0.25/4 and 2/32 (parC79); strain M16, 0.5/2 and 8/32 (parC83); strain Gyr-1207, 1.5/3 and 8/16 (gyrA); and strain MQ3A, 4/4 and 16/64 (parC and gyrA) . Both drugs were efficient with wild type-expPP, but only MFX was efficient with efflux-expPP . No bacterial reduction was observed for parC-expPPs due to mutants observed in 18 to 100% of animals, depending on the strain and the drug tested . These mutants showed unbound area under the concentration-time curve and MICs of from 50 to 164 for MFX . The in vivo pharmacodynamic boundaries of the MSW were different for MFX and LVX . We conclude that, after LVX or MFX treatment, mutants occur in vivo if there is a preexisting parC mutation, since the drug concentrations fall below the MPCs of these strains . Since the MPC determination cannot be routinely determined, these phenotypes or genotypes should be detected by simple tests to guide the therapeutic options.

Antimicrob Agents Chemother, 2004 May, 48(5), 1520 - 5
Variety of beta-lactamases produced by amoxicillin-clavulanate-resistant Escherichia coli isolated in the northeastern United States; Kaye KS et al.; This study analyzed the enzymatic basis and molecular epidemiology of amoxicillin-clavulanate-resistant Escherichia coli isolated by the microbiology laboratory of a United States tertiary care hospital . From October 1998 to December 1999, all E . coli isolates were screened for ampicillin-sulbactam resistance . Of 283 isolates that tested resistant to ampicillin-sulbactam, 69 unique patient isolates were also resistant to amoxicillin-clavulanate by disk diffusion testing (zone diameter </= 13 mm) . These amoxicillin-clavulanate-resistant E . coli isolates underwent agar dilution testing, pulsed-field gel electrophoresis, PCR analysis, and isoelectric focusing . The mean age of study patients was 52 years; 78% were female . Among the isolates, 12 were nosocomial (rate of amoxicillin-clavulanate resistance = 4.7%) and 57 were community acquired (rate of amoxicillin-clavulanate resistance = 2.8%) . No predominant strain was identified . By agar dilution testing, 67 isolates were nonsusceptible (39 resistant and 28 intermediate) to amoxicillin-clavulanate and 37 were piperacillin-tazobactam resistant but only 8 were ceftazidime resistant (ceftazidime MIC >/= 32 micro g/ml) . Two isolates were susceptible to amoxicillin-clavulanic acid by agar dilution, although they were resistant by disk diffusion testing . The distribution of beta-lactamases was as follows: the TEM type alone was found in 52 isolates, the AmpC type was found in 4 isolates (2 identified as containing CMY-2), the TEM type and CMY-2 were found in 2 isolates, and the OXA type was found in 1 isolate . Also, there was one isolate with the TEM type and the SHV type and one with the TEM type and a second, unidentified enzyme . Among the isolates with TEM-type enzymes, two extended-spectrum beta-lactamase-producing isolates were identified but two isolates with inhibitor-resistant TEM (IRT) enzymes (one with TEM-34 {IRT-6} and the other with a novel enzyme {tentatively assigned the designation TEM-122}) were more interesting.

J Nat Prod, 2004 Apr, 67(4), 689 - 92
Isolation and structure elucidation of a novel antimalarial macrocyclic polylactone, menisporopsin A, from the fungus Menisporopsis theobromae; Chinworrungsee M et al.; An antimalarial macrocyclic polylactone, menisporopsin A (1), was isolated from a cell extract of the seed fungus Menisporopsis theobromae . The structure of 1 was elucidated on the basis of spectroscopic analysis and chemical transformations, with the absolute configuration established by application of the modified Mosher method and by using chiral HPLC . Menisporopsin A (1) possesses an unprecedented residue, 2,4-dihydroxy-6-(2,4-dihydroxy-n-pentyl)benzoic acid . This compound exhibited antimalarial activity, with an IC(50) value of 4.0 microg mL(-1), and antimycobacterial activity (MIC value of 50 microg mL(-1)).

J Nat Prod, 2004 Apr, 67(4), 598 - 603
Mycobacterium tuberculosis growth inhibition by constituents of Sapium haematospermum; Woldemichael GM et al.; Four novel compounds consisting of two new pimaranes, lecheronol A (1) and lecheronol B (2), an acylated cycloartane, 3-O-beta-lauroyl-cycloart-(23E)-en-25-ol (10), and a highly oxygenated novel chalconoid, alpha,beta,3,4,5,2',4',6'-octahydroxydihydrochalcone (12), were isolated along with seven known triterpene derivatives and three flavonol glucosides from Mycobacterium tuberculosis growth-inhibiting fractions of the CH(2)Cl(2)/MeOH (1:1) extract of the aerial parts of Sapium haematospermum . Compounds 1, 3 (3 alpha-hydroxyolean-12-ene), 8 {3 alpha-hydroxylup-20(29)-en}, and 9 (cycloartanol) were found most active, with MIC values of 4, 12.2, 13.4, and 8 microg/mL, respectively . Cytotoxicity tests in Vero cells for compounds 1, 3, 8, and 9 gave IC(50) values of 104.8, 127.2, 127.2, and 102.4 microg/mL, respectively.

J Med Microbiol, 2004 May, 53(Pt 5), 403 - 6
Micro-broth dilution method with air-dried microplate for determining MICs of clarithromycin and amoxycillin for Helicobacter pylori isolates; Kobayashi I et al.; MICs of clarithromycin and amoxycillin for 253 isolates of Helicobacter pylori were measured by an air-dried microplate method and compared with the results obtained by the agar plate dilution method . The air-dried microplate method is performed by coating each well of a 96-well microplate with the test antibiotic and air-drying it . There were no marked differences between the air-dried microplate method and agar plate dilution methods in the MIC(50) and MIC(90) values or MIC ranges of clarithromycin obtained for the 253 isolates of H . pylori . More specifically, the MICs of clarithromycin for 114 (45.1 %) of the 253 isolates were the same by the air-dried microplate method as the agar plate dilution method, and the differences in the MICs of clarithromycin for a further 114 isolates (45.1 %) varied within one twofold dilution . The MICs of amoxycillin by the former method were in close agreement with the MICs obtained by the latter method: MICs of amoxycillin for 199 (78.7 %) of the 253 isolates were the same by both methods, and the differences in the MICs of amoxycillin for 42 isolates (16.6 %) varied within one twofold dilution . These results indicate that the air-dried microplate method is a useful method for determination of MICs, because the results obtained were in close agreement with those obtained by the standard agar plate dilution method . The air-dried microplate method is, therefore, a convenient and reliable method for determining the MICs of clarithromycin and amoxycillin for H . pylori isolates.

Br J Ophthalmol, 2004 May, 88(5), 619 - 25
Doxycycline-a role in ocular surface repair; Smith VA et al.; BACKGROUND/AIMS: Doxycycline is a broad spectrum antibiotic that chelates metal ions and is frequently used as part of the treatment of ocular surface diseases . Its therapeutic value has been ascribed to an ability to inhibit matrix metalloproteinase (MMP) activity and both MMP and IL-1 synthesis . The aim of this study was to evaluate the role of doxycycline as an inhibitor of corneal MMPs and assess its contribution to ocular surface repair mechanisms . METHODS: Corneal epithelial cell and keratocyte cultures were grown to confluence and incubated with IL-1alpha, LPS, doxycycline, or doxycycline and LPS in serum free medium for 4 days . The cells were either harvested and assayed for caspase-3 activity or stained with either AE5 or antivimentin antibodies . Media samples were concentrated and assayed for MMP activity by zymography or using a fluorigenic substrate . ELISA was used to quantify IL-1alpha, MMPs -1,-2,-3,-9, and TIMPs -1 and -2 . RESULTS: IL-1alpha and LPS had no effect on MMP/TIMP production by cultured corneal epithelial cells and keratocytes . Corneal MMP-2 inhibition by doxycycline was partially {Ca(2+)} dependent but irreversible . At the minimum inhibitory concentration, 100 micro m, doxycycline had no apparent effect on MMP and TIMP production, but ultimately caused the death of keratocytes and some of the epithelial cells that detached from their basement membrane . Caspase-3 activity was not detected in dead or dying keratocytes . The mechanism of cell death in cultured corneal epithelial cells was not caspase-3 related apoptosis as the activity of this enzyme, normally detectable, was lost . The epithelial cells that survived doxycycline treatment did not bind antivimentin antibody and compared with controls, reacted less with the AE5 antibody . They were probably transient amplifying cells . CONCLUSIONS: Doxycycline irreversibly inhibits corneal MMP-2 activity by chelating the metal ions that are catalytically and structurally essential . Corneal MMP/TIMP production in vitro is not modulated by IL-1alpha, LPS, or doxycycline . The therapeutic value of doxycycline may depend upon its effective concentration at the ocular surface and probably relates to its chelating properties.

Am J Gastroenterol, 2004 Apr, 99(4), 676 - 80
Association of MHC class I related gene B (MICB) to celiac disease; Gonzalez S et al.; BACKGROUND AND AIMS: Celiac disease (CD) is an enteropathic disorder characterized by strong association with HLA-DQ2 . Our aim was to investigate whether MICB, a gene located in the MHC class I region, may contribute to CD susceptibility . PATIENTS AND METHODS: Total of 133 CD patients, previously reported to be associated with MICA-A5.1, and 116 controls were initially analyzed . Twenty-eight additional DQ2-negative CD patients were also studied . MICB was typed by PCR using sequence-specific primers . HLA-B, -DRB1, -DQA1, -DQB1, and MICA were also typed . RESULTS: The allele MICB0106 was strongly associated with CD (pc < 0.000001, odds ratio (OR) = 5.6, 95% confidence interval (CI) = 3.1-10.1) and it was overrepresented in atypical patients compared with typical ones (pc = 0.04, OR = 2.9, CI = 1.4-6.1) . MICB0106 was part of DR3-DQ2 haplotype (B8-MICA-A5.1-MICB0106-DR3-DQ2), and consequently a strong linkage disequilibrium between MICB0106 with DQ2 (lambdas = 1) and MICA-A5.1 (lambdas = 0.55) was found . To analyze whether the association of MICB is independent of this haplotype, its association was also studied in DQ2-negative patients (n = 46) . DQ8 (28%vs 9%, p = 0.0085, pc = NS) and MICB0104 (52%vs 30%, p = 0.01, pc = NS) were increased in DQ2-negative patients . MICA-A5.1 was significantly increased in atypical patients (p(c)= 0.001, OR = 6.4, CI = 2.2-18.4), and this association was independent of DQ2 and DQ8 (pc = 0.02, OR = 2.6, CI = 1.1-6.1) . CONCLUSIONS: The expression of MIC genes on enterocytes under stressful conditions and their function as ligands of intraepithelial gammadelta and CD8 T cells, together with the data presented here suggest a potential role of MIC genes in the pathogenesis of CD.

J Pediatr Gastroenterol Nutr, 2004 Apr, 38(4), 426 - 9
Longevity of balloon-stabilized skin-level gastrostomy device; Michaud L et al.; OBJECTIVE: The gastrostomy button is increasingly used in patients requiring long-term enteral feeding . No data are available addressing the longevity of balloon-type gastrostomy buttons . The aim of this study was to evaluate the longevity of balloon-type gastrostomy buttons in children and the cause of the button removal . PATIENTS AND METHODS: During a period of 2.5 years, 165 gastrostomy (Mic-Key, Medical Innovations Corporation, Draper, Utah, U.S.A.) buttons were inserted in 84 children . In all patients, the button replaced a standard tube gastrostomy . The first button was usually inserted under general anesthesia at least 2 months after the insertion of a surgical gastrostomy tube . Additional replacement was performed without sedation . RESULTS: Mean longevity of balloon-type gastrostomy buttons was 5 months (range, 14 days-14 months) . Causes of button removal were: inner balloon rupture (n = 101, 61%); leak around button or deflation of the balloon (n = 23, 14%); accidental removal (n = 20, 12%); device damage (n = 5), and obstruction (n = 3) . When considering the subgroup with inner balloon rupture, the longevity of the button was 6 months (range, 1-13 months) . No correlation was found between duration of gastrostomy button and underlying disease, age of the patient, or the use of antisecretory drugs . CONCLUSION: Although the Mic-Key button has the advantage of an easy insertion, its major limitation is the high frequency of inner balloon rupture, which was the primary main reason for removal.

Farmaco, 2004 Apr, 59(4), 279 - 88
Synthesis and antimycobacterial activity of 1,2,4-triazole 3-benzylsulfanyl derivatives; Klimesova V et al.; Series of 3-benzylsulfanyl derivatives of 1,2,4-triazole and 4-methyl-1,2,4-triazole were synthesized by alkylation of starting triazole-3-thiol with appropriately substituted benzyl halide . All members of the set were evaluated for in vitro antimycobacterial activity against Mycobacterium tuberculosis, M . avium, and two strains of M . kansasii . The activities were expressed as the minimum inhibitory concentration . The compounds exhibited only a moderate or slight antimycobacterial activity . Minimum inhibitory concentrations fall into a range of 32->1000 micromol/l . The most active substances bear two nitro groups or a thioamide group on the benzyl moiety . As regards the cytotoxicity effect, the evaluated compounds can be considered as moderately toxic.

Int J Antimicrob Agents, 2004 Apr, 23(4), 408 - 11
The vitro efficacy of beta-lactam and beta-lactamase inhibitors against multidrug resistant clinical strains of Mycobacterium tuberculosis; Dincer I et al.; In vitro activity of beta-lactam antibiotics and their beta-lactamase inhibitor combinations were evaluated for their activity against clinical isolates of Mycobacterium tuberculosis and M . tuberculosis H37Ra . Agar dilution, the BACTEC 460 system and beta-lactamase activity tests were used . Results using the BACTEC 460 and enzyme activity tests showed the best beta-lactamase inhibitor for M . tuberculosis H37Ra to be clavulanic acid . Cefazolin-clavulanic acid gave the lowest minimal inhibitory concentration (MIC) values using dilution tests and M . tuberculosis H37Ra . beta-Lactam antibiotics were combined with clavulanic acid and tested for in vitro activity against 50 selected multidrug-resistant (MDR) and 50 susceptible clinical isolates . Seventy-four percent of the isolates were inhibited by cefazolin-clavulanic acid combination . These results suggested that an appropriate combination of beta-lactam and beta-lactamase inhibitors might be useful in the treatment of tuberculosis due to multidrug-resistant strains.

Bioorg Med Chem, 2004 May 1, 12(9), 2225 - 38
Optically active antifungal azoles: synthesis and antifungal activity of (2R,3S)-2-(2,4-difluorophenyl)-3-(5-{2-{4-aryl-piperazin-1-yl}-ethyl}-tetrazol-2-yl/1-yl)-1-{1,2,4}-triazol-1-yl-butan-2-ol; Upadhayaya RS et al.; A series of (2R,3S)-2-(2,4-difluorophenyl)-3-(5-{2-{4-aryl-piperazin-1-yl}-ethyl}-tetrazol-2-yl)-1-{1,2,4}-triazol-1-yl-butan-2-ol (11a-n) and (2R,3S)-2-(2,4-difluorophenyl)-3-(5-{2-{4-aryl-piperazin-1-yl}-ethyl}-tetrazole-1-yl)-1-{1,2,4}-triazol-1-yl-butan-2-ol (12a-n) has been synthesized . The antifungal activity of compounds was evaluated by in vitro agar diffusion and broth dilution assay . Compounds 11d and its positional isomer 12d having 3-trifluoromethyl substitution on the phenyl ring of piperazine demonstrated significant antifungal activity against variety of fungal cultures (Candida spp . C . neoformans and Aspergillus spp.) . The compound 12d showed MIC value of 0.12 microg/mL for C . albicans, C . albicans V-01-191A-261 (resistant strain); 0.25 microg/mL for C . tropicalis, C . parapsilosis ATCC 22019 and C . krusei and MIC value of 0.5 microg/mL for C . glabrata, C . krusei ATCC 6258, which is comparable to itraconazole and better than fluconazole . Further, compound 11d showed significant activity (MIC; 0.25-0.5 microg/mL) against Candida spp . and strong anticryptococcal activity (MIC; 0.25 microg/mL) against C . neoformans.

Chest, 2004 Apr, 125(4), 1400 - 5
Efficacy of mechanical insufflation-exsufflation in medically stable patients with amyotrophic lateral sclerosis; Sancho J et al.; OBJECTIVE: To determine under what circumstances the use of mechanical insufflation-exsufflation (MI-E) can generate clinically effective expiratory flows for airway clearance (> 2.7 L/s) for clinically stable patients with amyotrophic lateral sclerosis (ALS) . MATERIALS AND METHOD: Twenty-six consecutive patients with ALS were studied, 15 with severe bulbar dysfunction . Using a pneumotachograph and with the aid of an oronasal mask, we measured FVC, FEV(1), peak cough flow (PCF), maximum insufflation capacity (MIC), PCF generated from a maximum insufflation MIC (PCFMIC), and PCF generated by MI-E (PCFMI-E) . MI-E was delivered at +/- 40 cm H(2)O . Maximum inspiratory pressure (PImax) and maximum expiratory pressure (PEmax) at the mouth were also measured . RESULTS: Although both groups had a similar time from ALS symptom onset to diagnosis, statistical differences (p < 0.05) were found between nonbulbar and bulbar patients in lung function and cough capacity parameters: FVC, 2.58 +/- 1.24 L vs 1.62 +/- 0.74 L; FEV(1), 2.26 +/- 1.18 L vs 1.54 +/- 0.69 L; PImax, - 93.45 +/- 47.47 cm H(2)O vs - 3.64 +/- 25.07 cm H(2)O; PEmax, 140.45 +/- 75.98 cm H(2)O vs 69.93 +/- 32.14 cm H(2)O; MIC, 3.02 +/- 1.22 L vs 1.97 +/- 0.75 L; PCF, 5.91 +/- 2.55 L/s vs 3.42 +/- 1.44 L/s; PCFMIC, 6.68 +/- 2.71 L/s vs 4.00 +/- 1.48 L/s; and PCFMI-E, 4.34 +/- 0.82 L/s vs 3.35 +/- 0.77 L/s . Four patients with bulbar dysfunction and MIC > 1 L had PCFMI-E < 2.7 L/s . The receiver operating characteristic (ROC) curve analysis showed PCFMIC of <or= 2.7 L/s predicting those patients with PCFMI-E < 2.7 L/s . The ROC curve analysis showed PCFMIC > 4 L/s predicting those patients with PCFMIC greater than PCFMI-E . CONCLUSION: MI-E is able to generate clinically effective PCFMI-E (> 2.7 L/s) for stable patients with ALS, except for those with bulbar dysfunction who also have a MIC > 1 L and PCFMIC <2.7 L/s who probably have severe dynamic collapse of the upper airways during the exsufflation cycle . Clinically stable patients with mild respiratory dysfunction and PCFMIC > 4 L/s might not benefit from MI-E except during an acute respiratory illness.

Mycoses, 2004 Apr, 47(3-4), 104 - 14
Pharmacokinetic investigation of oral itraconazole in stratum corneum level of tinea pedis; Morimoto K et al.; In the present study, the authors administered 100 mg itraconazole (ITCZ) twice daily for a period of 1 week to six patients with hyperkeratotic type tinea pedis, and examined its efficacy, safety profile, and usefulness . ITCZ concentration in stratum corneum was also measured to examine the mobility of the drug into the affected site of planta pedis . ITCZ concentration in the stratum corneum of the affected part was first detected at 1 week after the completion of administration, gradually increased over time, and peaked at 3 weeks, with the sum of ITCZ and hydroxyitraconazole (OH-ITCZ) amounting to 163.7 ng g(-1) on a average . It then gradually decreased to a total sum of 10.3 ng g(-1) on average at 8 weeks following the completion of administration . When compared with the geometric mean minimum inhibitory concentration (MIC) of ITCZ against fresh clinical isolates of dermatophytes (Trichophyton rubrum) (0.06 microg ml(-1)), the stratum corneum ITCZ concentration in this study was 2.1-fold of the geometric mean MIC at 2 weeks following the completion of administration, and 2.4-fold at 4 weeks . Although ITCZ does not produce therapeutic effectiveness (fungistasis) during the period of administration, it starts appearing at 2 weeks after the completion of administration, and after it peaks out at 3-4 weeks, clinical symptoms started improving . These results suggest that satisfying effects can be achieved in a short-term oral ITCZ at a dose of 100 mg twice daily for a period of 1 week in cases of hyperkeratotic type tinea pedis.

Microbiology, 2004 Apr, 150(Pt 4), 853 - 64
The MspA porin promotes growth and increases antibiotic susceptibility of both Mycobacterium bovis BCG and Mycobacterium tuberculosis; Mailaender C et al.; Porins mediate the diffusion of hydrophilic solutes across the outer membrane of mycobacteria, but the efficiency of this pathway is very low compared to Gram-negative bacteria . To examine the importance of porins in slow-growing mycobacteria, the major porin MspA of Mycobacterium smegmatis was expressed in Mycobacterium tuberculosis and Mycobacterium bovis . Approximately 20 and 35 MspA molecules per microm(2) cell wall were observed in M . tuberculosis and M . bovis BCG, respectively, by electron microscopy and quantitative immunoblot experiments . Surface accessibility of MspA in M . tuberculosis was demonstrated by flow cytometry . Glucose uptake was twofold faster, indicating that the outer membrane permeability of M . bovis BCG to small and hydrophilic solutes was increased by MspA . This significantly accelerated the growth of M . bovis BCG, identifying very slow nutrient uptake as one of the determinants of slow growth in mycobacteria . The susceptibility of both M . bovis BCG and M . tuberculosis to zwitterionic beta-lactam antibiotics was substantially enhanced by MspA, decreasing the minimal inhibitory concentration up to 16-fold . Furthermore, M . tuberculosis became significantly more susceptible to isoniazid, ethambutol and streptomycin . Fluorescence with the nucleic acid binding dye SYTO 9 was 10-fold increased upon expression of mspA . These results indicated that MspA not only enhanced the efficiency of the porin pathway, but also that of pathways mediating access to large and/or hydrophobic agents . This study provides the first experimental evidence that porins are important for drug susceptibility of M . tuberculosis.

Clin Cancer Res, 2004 Apr 1, 10(7), 2386 - 92
Serum macrophage inhibitory cytokine 1 as a marker of pancreatic and other periampullary cancers; Koopmann J et al.; PURPOSE: Patients with pancreatic ductal adenocarcinoma usually present with advanced-stage disease and a dismal prognosis . One effective strategy likely to improve the morbidity and mortality from pancreatic cancer would be the identification of accurate, noninvasive diagnostic markers that would enable earlier diagnosis of symptomatic patients and earlier detection of cancer in asymptomatic individuals at high risk for developing pancreatic cancer . In this study, we evaluated serum macrophage inhibitory cytokine-1 (MIC-1) as a marker of pancreatic cancer . EXPERIMENTAL DESIGN: MIC-1 expression in primary pancreatic cancers, intraductal papillary mucinous neoplasms, and pancreatic cancer cell lines was determined using the National Center for Biotechnology Information serial analysis of gene expression database, oligonucleotide microarrays analysis, in situ hybridization, and immunohistochemistry . Serum MIC-1 levels were determined by ELISA in 80 patients with pancreatic adenocarcinomas, in 30 patients with ampullary and cholangiocellular carcinomas, in 42 patients with benign pancreatic tumors, in 76 patients with chronic pancreatitis, and in 97 healthy control subjects . The diagnostic performance of serum MIC-1 as a marker of pancreatic cancer was compared with that of serum CA19-9 . RESULTS: Oligonucleotide microarray and serial analysis of gene expression data demonstrated that MIC-1 RNA levels were higher in primary pancreatic cancers, intraductal papillary mucinous neoplasms, and pancreatic cancer cell lines than in nonneoplastic pancreatic ductal epithelium . MIC-1 expression was localized to the malignant epithelium in pancreatic adenocarcinomas by in situ hybridization . MIC-1 protein was expressed in 14 of 16 primary pancreatic adenocarcinomas (88%) by immunohistochemistry and was also expressed in some pancreata affected by pancreatitis but not in normal pancreas . Serum MIC-1 levels were significantly higher in patients with pancreatic ductal adenocarcinoma (mean +/- SD, 2428 +/- 2324 pg/ml) and in patients with ampullary and cholangiocellular carcinomas (2123 +/- 2387 pg/ml) than in those with benign pancreatic neoplasms (940 +/- 469 pg/ml), chronic pancreatitis (1364 +/- 1236 pg/ml), or in healthy controls (546 +/- 262 pg/ml) . An elevated serum MIC-1 (defined as 2 SD above the mean for healthy controls) performed as well as CA19-9 (area under the receiver operating characteristic curve, 0.81 and 0.77, respectively), and the combination of MIC-1 and CA19-9 significantly improved diagnostic accuracy (P < 0.05; area under the receiver operating characteristic curve, 0.87; sensitivity, 70%; specificity, 85%) . CONCLUSION: Serum MIC-1 measurement can aid in the diagnosis of pancreatic adenocarcinoma.

Anal Bioanal Chem, 1996 Mar, 354(5-6), 750 - 5
Vitality fertilization of Scots pine stands growing along a gradient of heavy metal pollution: short-term effects on microbial biomass and respiration rate of the humus layer; Fritze H et al.; In 1992 forest vitality fertilization experiments were established on a heavy metal deposition gradient with four treatments in three replications at distances of 0.5, 4 and 8 km from a Cu-Ni smelter in order to estimate their impact on the disturbed forest ecosystem . The increase in Cu concentration in the humus (F/H) layer of the Calluna site type Scots pine (Pinus sylvestris) stands from ca . 300 to 8000 mg kg(-1) d.m . (dry matter) along the 8 km long transect towards the smelter resulted in declining soil microbial biomass and soil respiration activity . Three independent measurements of microbial biomass: C(mic)-FE (fumigation extraction), C(mic)-SIR (substrate induced respiration), and ATP have been used together with an indicator of fungal biomass (ergosterol) and microbial activity (soil respiration) . Within this Cu pollution range, all the measured microbial biomass levels declined to 10%-28% of the control plot values and activity assessed by respiration was lowered to 16% . Liming has increased the C(mic)-SIR and respiration rate . Treatments with test fertilizer, made from grounded apatite, did not result in different microbial biomass and respiration rate values compared to the respective controls along the whole gradient . Nitrogen + lime treatments resulted in similar changes to lime alone . No changes, as compared to the respective control, could be detected with nitrogen fertilization at the less polluted end of the gradient.

Prostate, 2004 Jun 1, 59(4), 351 - 6
Down-regulation of macrophage inhibitory cytokine-1/prostate derived factor in benign prostatic hyperplasia; Kakehi Y et al.; BACKGROUND: Macrophage inhibitory cytokine-1 (MIC-1) is a member of transforming growth factor-beta/bone morphogenetic protein (BMP) superfamily . Despite its potential role in prostatic regulation, little is known about its biological activity . METHODS: Expression profiling using 42K Affymetrix HuGeneFL array was conducted to compare symptomatic benign prostatatic hyperplasia (BPH), histological BPH without symptoms, and normal prostate samples from donors . MIC-1 gene expression was analyzed by RT-PCR in pure culture of prostate epithelial and stromal cells, and prostate cancer cells, LNCaP, PC-3, DU-145 . Influence of androgens on MIC-1 expression in LNCaP cells was analyzed by Northern blot . Enhancement of promoter activity of MIC-1 by androgens was examined using reporter assays . RESULTS: In contrast to normal prostates, MIC-1 gene was down-regulated in BPH samples with symptoms and histological BPH obtained from cystoprostatectomy specimens (P < 0.005 and P < 0.01, respectively) . Expression level of MIC-1 in androgen-sensitive LNCaP cells was high and enhanced by androgens, whereas in the androgen-insensitive PC-3 and DU-145 cells the expression level was low . An 11 kb promoter region of MIC-1 gene was identified to be 6- to 12-fold activated by androgens . CONCLUSIONS: Down-regulation of MIC-1 may play a role in the development of BPH . MIC-1 is positively regulated by androgens, but other regulatory factors remain unclear .

Eur J Obstet Gynecol Reprod Biol, 2004 Apr 15, 113(2), 229 - 33
Laser CO(2) conization: a safe mode of treating conservatively microinvasive carcinoma of the uterine cervix; Diakomanolis E et al.; OBJECTIVE: To evaluate the outcome of conservative treatment by laser CO(2) conization, for the management of microinvasive carcinoma of the uterine cervix (MIC) . Study design: From 1990 to 1999, 90 women with the diagnosis of MIC were treated in the Gynecological Oncology Unit of "Alexandra" Hospital . Final diagnosis of MIC was based on cervical conization as well as simple and radical hysterectomy specimens . The cytological and colposcopical diagnoses prior to conization were reviewed . The cone specimen parameters examined by the pathologists were depth and width of invasion, lymph-vascular space invasion (LVSI) and surgical margins status . The modality used for all conizations, either primary or secondary, was the laser CO(2) under local anaesthesia . RESULTS: Diagnosis of MIC was made on cone biopsy in 73 women (81%), in simple hysterectomy in 10 (11%) and in radical hysterectomy specimens in 7 (8%) . From the patients that underwent conization, two (2.5%) were detected with LVSI . Five patients (7%) were found to have involved margins and from those, the majority was managed by a second conization . Mean follow-up time was 54 months (range: 30-110 months) . Four patients (6.6%) with recurrence were observed during follow-up, all of them with LSIL . No cases of invasive disease or HSIL were encountered . CONCLUSIONS: Laser CO(2) conization is a safe and effective mode of treatment for women suffering from MIC and wish to retain their fertility . However, this type of management should be advocated only in cases fulfilling the strict criteria for MIC as these have been defined by FIGO.

Diagn Microbiol Infect Dis, 2004 Apr, 48(4), 259 - 64
In vitro susceptibility testing of dermatophytes: comparison of disk diffusion and reference broth dilution methods; Karaca N et al.; A total of 56 strains belonging to 4 species of dermatophytes were tested against 10 antifungal drugs by using a modification of the NCCLS (M38-P) standard for filamentous fungi . The minimum inhibitory concentration (MIC) values obtained using the dilution method were compared with the diameters of growth inhibition zones using the disk diffusion method . The antifungals used were itraconazole, fluconazole, ketoconazole, miconazole, sulconazole, oxiconazole, bifonazole, griseofulvin, ciclopiroxolamine, and terbinafine . Relative to the other agents tested, terbinafine possessed the highest antifungal activity against all of the dermatophytes . In contrast, fluconazole was the least active drug . An increase of MIC values was accompanied by a decrease of growth inhibition zone diameter . The disk diffusion method of fungal susceptibility assessment yields data consistent with results obtained from the dilution method . The study suggests the potential value of the disk diffusion method as a convenient alternative method for testing the susceptibilities of dermatophytes.

Scand J Infect Dis, 2004, 36(2), 85 - 92
The fluoroquinolones: how long will their utility last?
Bakken JS.
Injudicious use of the fluoroquinolones may result in treatment failure, increased patient morbidity, increased health care cost, and possible patient fatality . Fluoroquinolone-resistant bacteria may also adversely impact the microbiological environment in the hospital, the local community and eventually large geographical regions . Fluoroquinolone resistance develops in a stepwise fashion, and current susceptibility testing methods and recommended MIC susceptible breakpoint values for the United States may fail to identify some bacteria that are resistant due to first step mutations at the fluoroquinolone target site gene sequences . C-8 methoxy- fluoroquinolone compounds are more active against resistant bacteria than the older compounds . Fluoroquinolone resistance relates directly to human and veterinary usage and emerging bacterial resistance poses the single greatest threat to the future survival of the fluoroquinolone drugs as an antibiotic class.

Clin Microbiol Infect, 2004 Apr, 10(4), 315 - 21
Laboratory handling of Helicobacter pylori critically influences the results of in-vitro metronidazole resistance determination; Henriksen TH et al.; In-vitro metronidazole resistance rates of Helicobacter pylori determined by Etest are high, and the predictive value of metronidazole resistance is low . It was hypothesised that altered laboratory methods could reduce the overestimation of resistance and improve the predictive value of the Etest . Pre-treatment isolates (n = 150) of H . pylori from 150 patients were investigated by Etest with incubation for 72 h . Treatment with metronidazole, tetracycline and bismuth for 10 days failed to eradicate H . pylori in 23 patients . After isolate storage for 3 years, resistance determination results by agar dilution and Etest, with incubation for 72 and 31 h, were compared . The rate of metronidazole resistance was reduced significantly during storage, and instability of resistance was associated significantly with treatment outcome . Isolates that retained in-vitro resistance had significantly (p 0.008) higher treatment failure rates (n = 13; 42%) than isolates that lost resistance (n = 3; 9%) . The reproducibility achieved by dual testing with agar dilution and Etest was 41% and 70% for +/- 1 and +/- 2 log2 dilutions, respectively, after incubation for 72 h, and 85% and 92%, respectively, after incubation for 31 h . Thus, the predictive value was improved from 25% to 50% by the altered laboratory conditions (p 0.04) . MIC values of 2-8 mg/L signified an intermediate risk of treatment failure.

Polim Med, 2003, 33(4), 29 - 42
Effect of natural anionic polymers on the release rate and local concentration of metronidazole at pH near the natural gastric environment; Musial W et al.; The aim of the study was to evaluate initially the applicability of certain natural polymers, as metronidazole binding agents in the treatment of ulcerative lesions in the gastric mucous membrane . The investigation involved systems containing metronidazole in combination with sodium alginate, citrus pectin and citrus-apple pectin . The rate of metronidazole release from the investigated preparations was evaluated after acidification to pH in gastric environment . Constant rates were determined, which were from 0.25 h-1 to 0.58 h-1 as well as half-release time, which were from 1.20 h to 2.82 h . Also, dynamic viscosity of the investigated systems was assessed--the dynamic viscosity of the gelatinous form with 2% content of sodium alginate increased by three orders of magnitude under the influence of acid pH, i.e . from about 160 cPa.s to about 30,000 cPa.s . Similarly, the viscosity of the preparation containing 4% of citrus-apple pectin increased after acidification from about 140 cPa.s to almost 40,0000 cPa.s . The use of sodium alginate and citrus-apple pectin as metronidazole carriers may prolong the time of action of the drug on pathologically changed gastric mucosa . The fraction of metronidazole which is in immediate contact with the pathologically changed surface is in concentration exceeding significantly MIC and MBC at usual oral doses.

Prenat Diagn, 2004 Mar, 24(3), 224 - 6
Maternal serum and amniotic fluid levels of macrophage inhibitory cytokine 1 in Down syndrome and chromosomally normal pregnancies; Wallace EM et al.; OBJECTIVES: To measure maternal serum and amniotic fluid levels of macrophage inhibitory cytokine-1 (MIC-1) in Down syndrome and normal pregnancies, assessing the utility of MIC-1 as a prenatal marker of Down syndrome . METHODS: Stored serum from 64 Down syndrome and 399 control pregnancies, collected at 8 to 17 weeks of pregnancy, and stored amniotic fluid from 17 Down syndrome and 53 controls, collected at 15 to 19 weeks of pregnancy, were retrieved for analysis . MIC-1 was measured using an established in-house ELISA, blinded to sample type . RESULTS: In maternal serum, MIC-1 levels are not altered in Down syndrome in either the first or second trimester . Levels, expressed as median (95% CI) multiples of the median (MoM), in the Down syndrome cases and controls were 1.07 (0.9-1.1) MoM and 1.0 (0.95-1.03) MoM respectively . In amniotic fluid, MIC-1 levels were significantly decreased compared to controls, 0.52 (0.44-0.64) MoM versus 1.0 (0.85-1.08) MoM (p < 0.0001) . CONCLUSION: MIC-1 is decreased in amniotic fluid but not in maternal serum in Down syndrome pregnancies . MIC-1 will not be useful as a prenatal marker of Down syndrome .

Folia Biol (Praha), 2004, 50(1), 21 - 3
Fluorescence-based automated fragment analysis of microsatellite polymorphism within the transmembrane region of the MIC-A gene; Novota P et al.; MHC class I chain-related genes (MIC) are located within the MHC class I region of chromosome 6 . Sequence analysis of the MIC-A gene showed a trinucleotide repeat (GCT) microsatellite polymorphism within the transmembrane region . So far, six alleles of the exon 5 of the MIC-A gene, which consist of 4, 5, 6, 9 and 10 repetitions of GCT, or five repetitions of GCT with an additional nucleotide insertion (GGCT), have been identified . Recent works support the findings that MIC-A is associated with several autoimmune diseases . In our work we present a modification of a method used for microsatellite polymorphism detection within the transmembrane region of the MIC-A gene . It is the ALFexpress fluorescence-based automated fragment analysis . We also present the frequencies of MIC-A exon 5 alleles found in the Czech population . We have identified five alleles of the transmembrane region of MIC-A, which comprise 4, 5, 6 and 9 repetitions or five repetitions with an additional nucleotide insertion . The most frequent allele was A5.1 (59.3%) and the less frequent was the allele A5 (20.0%) . No A7, A8 or A10 alleles were identified.

World J Gastroenterol, 2004 Apr 1, 10(7), 1075 - 7
New mutation points in 23S rRNA gene associated with Helicobacter pylori resistance to clarithromycin in northeast China; Hao Q et al.; AIM: To investigate the resistance rate of Helicobacter pylori (H pylori ) to clarithromycin, metronidazole, amoxicillin and tetracycline to guide clinical practice, and to study the mechanism of H pylori resistant to clarithromycin . METHODS: Thirty H pylori strains were isolated from the mucosa of peptic ulcer, gastric tumor and chronic gastritis patients, then the minimal inhibitory concentration (MIC) to clarithromycin, metronidazole, amoxicillin and tetracycline was evaluated by E-test method . The sequence analysis of PCR fragments was conducted in 23S rRNA gene of H pylori resistant to clarithromycin to get the resistance mechanism of the bacteria . RESULTS: Among 30 H pylori strains, 7 cases were resistant to clarithromycin, 12 to metronidazole, 2 to tetracycline and no strain was found to be resistant to amoxicillin . The resistance rates were 23.3%, 40%, 6.7% and 0%, respectively . Three new mutation points were found to be related to the clarithromycin resistance in H pylori isolates, which were G2224A, C2245T and T2289C . CONCLUSION: In northeast China, H pylori shows high resistance to metronidazole, while sensitive to amoxicillin . The mechanism of resistance to clarithromycin may be related to the mutation of G2224A, C2245T and T2289C in the 23S rRNA gene.

Int J Clin Pharmacol Ther, 2004 Mar, 42(3), 165 - 73
Amoxicillin/clavulanic acid (875/125): bioequivalence of a novel Solutab tablet and rationale for a twice-daily dosing regimen; Sourgens H et al.; A new amoxicillin/clavulanic acid tablet formulation (Solutab tablet, Forcid Solutab) containing amoxicillin/clavulanic acid (875/125) has been developed . The aim of the present study was to demonstrate bioequivalence between the new tablet formulation (test), taken as an intact tablet and after prior dispersal, versus the originator product viz . Augmentan film-coated tablet (875/125) used as reference . The study was performed in 48 healthy volunteers according to an open, single-dose, crossover design . Bioequivalence was demonstrated using Cmax and AUC(0-infinity) as primary parameters of evaluation for both amoxicillin and clavulanic acid with 90% confidence intervals of the ratios Solutab tablet/Augmentan within the range of 0.8-1.25 . The duration of the plasma concentration exceeding the amoxicillin minimal inhibitory concentration (MICs) was calculated using individual plasma concentration-time curves and compartmental analysis . The data showed that the bioavailability characteristics of the test tablet, taken intact or in dispersed form, and the reference tablets were very similar . The analysis, moreover, also confirmed the appropriateness of using a b.i.d . dosage regimen for both formulations, taking into account the pharmacodynamic breakpoint values for some major pathogens.

J Postgrad Med, 2004 Jan-Mar, 50(1), 17 - 20
Associations of antifolate resistance in vitro and point mutations in dihydrofolate reductase and dihydropteroate synthetase genes of Plasmodium falciparum; Biswas S; BACKGROUND: Antifolate antimalarials like sulfadoxine-pyrimethamine are used as second-line treatment for Plasmodium falciparum malaria patients who fail to respond to chloroquine . The efficacy of the sulfa-pyrimethamine combination in the treatment is also compromised by the development of resistance in the parasite . Resistance to these drugs has been shown to encode with point mutations in dihydrofolate reductase and dihydropteroate synthetase genes . SETTINGS: An experimental study . MATERIAL AND METHODS: Forty clinical isolates collected from different geographical locations in India were used to assess the relationships between resistance to sulfadoxine-pyrimethamine (SP) and mutations in P . falciparum dihydrofolate reductase (DHFR) and dihydropteroate synthetase (DHPS) . In vitro drug susceptibility and mutation-specific polymerase chain reaction (PCR) assays were also done . RESULTS: It was observed that a number of isolates possessed mutant genotypes and showed low sensitivity to SP in vitro . Of the 40 clinical isolates studied, 87.5% had DHFR and 15% had DHPS gene mutations . As observed from PCR results, 55( (22/40) presented double mutation of DHFR Arg-59 and Asn-108 and 32.5 % (13/40) had single mutant type allele of Asn-108 . Of the 40 isolates, 10 % (4/40) presented doubly mutated forms of DHPS Phe-436 and Thr-613 and single mutant type allele Gly-581 was detected in 5 % (2/40) isolates . Parasites carrying double or single mutant forms of DHFR/DHPS showed elevated minimum inhibitory concentration (MIC) values of both pyrimethamine (760-6754 nM; r=0.69) and sulfadoxine (108 - 540 micro M; r=0.87) when compared to sensitive and resistant strains . CONCLUSION: Though there was a correlation between molecular techniques and in vitro drug sensitivity profiles, the relevance of these findings to the clinical efficacy of SP combination drugs needs to be established by controlled clinical trials.

Antimicrob Agents Chemother, 2004 Apr, 48(4), 1197 - 203
Aspergillus fumigatus variant with decreased susceptibility to multiple antifungals; Balajee SA et al.; Isolates of Aspergillus fumigatus that demonstrate resistance to itraconazole (ITZ) have been described previously; however, the prevalence and clinical significance of ITZ resistance are not completely understood . In this study we assessed the ITZ susceptibilities of 128 A . fumigatus isolates that caused invasive infection in 82 stem cell transplant patients before and after the use of ITZ in our institution (study period, 1991 to 2000) . The MICs for 10 isolates obtained from seven patients were high, > or 1 microg/ml . The average ITZ MIC increased after institutional use of the drug began in 1995 . The majority of the isolates for which MICs were high (6 of 10) and one isolate for which the MIC was low (0.06 microg/ml) demonstrated an unusual phenotype, appearing as predominantly white colonies . For all seven atypical isolates, voriconazole MICs were high (> or = 2 microg/ml), and minimal effective concentrations of caspofungin were high (> or 4 microg/ml) . For two of the seven atypical isolates, amphotericin B MICs were high (> or 2 microg/ml) . The isolates appeared white due to slow sporulation; however, after prolonged incubations, the isolates sporulated with no difference in conidial color or conidiophore morphology compared with typical isolates . Randomly amplified polymorphic DNA-PCR patterns of these isolates were distinct compared with those of other A . fumigatus isolates . Sequencing of 18S rRNA genes confirmed that all were A . fumigatus; however, the mitochondrial cytochrome b gene sequences of all the atypical isolates were unique . These data suggest the potential presence of a genetically unique, poorly sporulating variant of A . fumigatus that demonstrates decreased susceptibilities to several antifungals.

Antimicrob Agents Chemother, 2004 Apr, 48(4), 1188 - 96
Efficacy, safety, and plasma pharmacokinetics of escalating dosages of intravenously administered ravuconazole lysine phosphoester for treatment of experimental pulmonary aspergillosis in persistently neutropenic rabbits; Petraitiene R et al.; Ravuconazole is a new antifungal triazole with broad-spectrum activity and a long half-life in plasma . We studied the antifungal efficacy, safety, and pharmacokinetics of ravuconazole lysine phosphoester in escalating dosages for the treatment of invasive pulmonary aspergillosis due to Aspergillus fumigatus in persistently neutropenic rabbits . Treatment groups consisted of rabbits treated with ravuconazole at 2.5 (RVC2.5), 5 (RVC5), and 10 (RVC10) mg/kg of body weight/day, rabbits treated with amphotericin B (AMB) at 1 mg/kg/day, or untreated controls . There was a dose-dependent reduction of pulmonary residual fungal burden (CFU per gram) in RVC5-, RVC10-, and AMB-treated rabbits in comparison to untreated controls (P < 0.01, P < 0.001, and P < 0.01, respectively) . These findings correlated with progressive galactomannan antigenemia in untreated controls and the RVC2.5-treated rabbits, a lower galactomannan index (GMI) in RVC5- and RVC10-treated rabbits, and a similarly low GMI in AMB-treated rabbits (P < 0.01) . Rabbits treated with RVC5, RVC10, and AMB also showed a reduction of organism-mediated pulmonary injury, as measured by infarct scores and lung weights, in comparison to untreated controls (P < 0.001) . These results were supported by decreased pulmonary infiltrates detected by computed tomography in RVC5- and RVC10-treated rabbits in comparison to untreated controls (P < 0.05) . Survival throughout the entire study was achieved in 95% of RVC5-treated rabbits (P < 0.001), 85% of RVC10-treated rabbits (P < 0.001), and 50% of AMB-treated rabbits (P < 0.05) in comparison to none of the untreated controls . Ravuconazole showed linear plasma pharmacokinetics and a large volume of distribution while maintaining concentrations in plasma above the MIC throughout the dosing interval . There was no evidence of hepatotoxicity or nephrotoxicity among ravuconazole-treated animals . Intravenously administered ravuconazole lysine phosphoester showed dose-dependent efficacy and an excellent safety profile for the treatment of invasive pulmonary aspergillosis in persistently neutropenic rabbits.

Gan To Kagaku Ryoho, 2004 Mar, 31(3), 346 - 50
{Ewing's sarcoma}; Goto T et al.; Ewing's sarcomas account for 6.8% of all primary malignant bone tumors and are probably a neurogenic, undifferentiated, high-grade malignancy, which usually affects the bones of children 5-15 years of age . Pain and swelling are the most common symptoms . Increase of CRP and erythrocyte sedimentation rate, leucocytosis, and anemia are frequently seen . Radiologically, they show permeative bone destruction on plain radiographs . When arising in the diaphysis of long bones, laminated, "onion-skin" periosteal reaction is seen . The tumor shows muscle density on CT, iso-signal intensity on T1-weighted MR images, and high signal intensity on T2-weighted MR images . Intramedullary invasion and skip lesions can be detected on MR images . Histologically, the tumor is uniformly composed of sheets of small round cells closely packed and without any matrix product . Glycogen granules are demonstrated in the cytoplasm by periodic acid-Schiff (PAS) and diastase reactions . Immunohistochemically, Ewing's sarcomas are positive for vimentin and MIC-2 gene product (CD99) . Reciprocal translocation, i.e., t(11;22) (q24;q12), is seen in the tumor cells . EWS/FLI-1 fusion gene can be demonstrated, which can be a complementary method in diagnosing this tumor . Because Ewing's sarcomas are chemosensitive and radiosensitive, they are treated by a combination of chemotherapy, surgery, and radiotherapy . Neoadjuvant chemotherapy consists of preoperative chemotherapy and postoperative chemotherapy . Preoperative chemotherapy aims at eradicating distant micrometastasis, reducing the primary tumor volume, and evaluating the efficacy of the chemotherapeutic agents . Surgery is performed as a local treatment by excising the tumor using the wide procedure . If surgery is impractical, curative radiotherapy is performed instead of excision . When surgery is performed without complete wide procedure, adjuvant radiotherapy is carried out to eradicate the residual tumor cells . Postoperative chemotherapy aims to eradicate the distant micrometastasis . Recently, myeloablative, high-dose chemotherapy followed by autologous bone marrow transplantation is being attempted for poor-prognosis patients and good results have been reported.

Anal Bioanal Chem, 1996 Jun, 355(3-4), 370 - 1
Gamma-ray spectrometric analysis of neutron irradiated golden sands; Daraban L et al.; A nondestructive method has been used for the qualitative and quantitative determination of elements such as Au, Cu, Ag in minerals and especially in golden sand . The induced radioactivity has been analyzed with a high resolution gamma spectrometer containing a Ge(Li) semiconductor detector type and a multichannel analyzer . The samples have been collected from the alluvia of the Mures and Somesul Mic river basins, proving the presence of gold and copper in small concentrations . These results have been correlated to previous data showing the existence of some gold mines in these zones . This nondestructive, rapid and highly sensitive method can be used for industrial processes {1} control and prospections . The use of a NaI(Tl) scintillation crystal for routine measurements of the (198)Au radioisotope type and the interferences by copper, silver and indium in the irradiated samples are discussed.

J Antimicrob Chemother, 2004 May, 53(5), 743 - 9 Epub 2004 Mar 24.
Effect of hypoxic conditions on in vitro susceptibility testing of amphotericin B, itraconazole and micafungin against Aspergillus and Candida; Warn PA et al.; OBJECTIVES: The aim of this study was to investigate the effect of hypoxic conditions on in vitro susceptibility testing of amphotericin B, itraconazole and micafungin against Aspergillus (four species) and Candida (six species) . METHODS: In vitro susceptibility tests were set up according to NCCLS M27-A2 and M38-A recommendations, but incubation atmospheres were either air plus 5% CO(2), 1% oxygen/5% CO(2)/94% nitrogen or 0.25% oxygen/5% CO(2)/94.75% nitrogen . RESULTS: In all Aspergillus species, the MIC of amphotericin B was reduced but the MFC remained unaltered with reduced oxygen . The MICs and MFCs of itraconazole and micafungin were unaltered in hypoxic conditions but interpretation of the MIC was much simpler for micafungin with 1% and 0.25% oxygen . Against Candida, conditions modelling hypoxia had little effect on the MICs and MFCs of any of the agents . CONCLUSIONS: This simple adaptation of susceptibility testing may have important consequences for understanding how antifungal drugs work and for endpoint reading.

J Nat Prod, 2004 Mar, 67(3), 485 - 6
Lakoochins A and B, new antimycobacterial stilbene derivatives from Artocarpus lakoocha; Puntumchai A et al.; Two new stilbene derivatives, lakoochins A (1) and B (2), were isolated from the roots of Artocarpus lakoocha . The structures of 1 and 2 were elucidated by analysis of their spectral data . Lakoochins A (1) and B (2) exhibited antimycobacterial activity with the respective MIC values of 12.5 and 50 microg/mL . While 1 was cytotoxic against the BC (breast cancer) cell line (IC(50) 6.1 microg/mL) but inactive (at 20 microg/mL) toward KB (nasopharyngeal carcinoma) cells, compound 2 possessed cytotoxicity against the BC and KB cell lines with IC(50) values of 3.1 and 6.1 microg/mL, respectively.

J Ethnopharmacol, 2004 Mar, 91(1), 37 - 42
In vitro antitrypanosomal activity of ethnopharmacologically selected Beninese plants; Hoet S et al.; The in vitro antitrypanosomal activity of methylene chloride, methanol and aqueous extracts of the leaves and twigs of five plant species traditionally used in Benin for the treatment of sleeping sickness were evaluated on Trypanosoma brucei brucei and their selectivity was analysed on Leishmania mexicana mexicana and J774 macrophage-like murine cells . The results showed that the four most active extracts had MIC values < or =19 microg/ml (Hymenocardia acida twig and leaf, Strychnos spinosa leaf, Trichilia emetica leaf methylene chloride extracts) . All these extracts had a lower activity on L . m . mexicana and J774 cells . Determination of the IC50 values of the methylene chloride leaf extracts on two strains of trypanosomes (T . b . brucei and T . b . rhodesiense) and two mammalian cell lines (L6 and J774 cells) showed that all extracts possessed some antitrypanosomal activity with IC50's ranging from 1.5 to 39 microg/ml . All were also toxic to the mammalian cells, but usually with higher IC50's . The only exception was the S . spinosa methylene chloride leaf extract which had no toxicity on J774 cells . Although tannins have been identified in most of the species studied, they could not be detected in the most active extracts, just as alkaloids . The presence of flavonoids and quinones may at least in part explain the observed activities of some of the active extracts.

J Ethnopharmacol, 2004 Mar, 91(1), 21 - 4
Antidermatophytic activity of extracts from Psoralea corylifolia (Fabaceae) correlated with the presence of a flavonoid compound; Rajendra Prasad N et al.; Extracts obtained from seeds of Psoralea corylifolia showed several degrees of antifungal activity against Trichophyton rubrum, Trichophyton mentagrophytes, Epidermophyton floccosum and Microsporum gypseum by the disc diffusion method on a Sabouraud dextrose agar (SDA) medium . Methanol extract of the seeds at 250 microg exhibited the maximum inhibition with a halo of 28 mm diameter . Six different bands were obtained when the methanol extract was subjected to TLC . 13C NMR and Mass spectra revealed that the active compound would be a flavonoid, 4'-methoxy flavone . MIC of the active compound along with standard miconazole was carried out using tube dilution technique.

J Colloid Interface Sci, 2004 Apr 15, 272(2), 253 - 61
Effect of alcohol addition to the aqueous phase on the thermal effects of micellization of cationic benzyldimethyldodecylammonium bromide and its adsorption onto porous and nonporous silicas; Benalla H et al.; Titration calorimetry has been used to study the effect of the addition of two primary alcohols, 1-butanol and 1-heptanol, to the aqueous phase on the thermal effects of micellization of benzyldimethyldodecylammonium bromide (BDDAB) as well as its adsorption onto nonporous Spherosil XO15M and onto porous aluminosilicate SiAl32d22 possessing uniformly sized mesopores . A linear decrease of the critical micelle concentration (CMC) of the cationic surfactant with the additive content was inferred from the specific conductivity measurements . All adsorption and calorimetry experiments were carried out at 298 K and at a fixed alcohol content (0.01 mol kg(-1)) either in deionized water or in a 0.01 mol kg(-1) NaBr solution . Dilution calorimetry measurements allowed determination of the cumulative molar enthalpy changes and a new analysis of these data was proposed to calculate easily the enthalpy of micellization per mole of BDDAB, Delta(mic)h, and the CMC value . The alcohol addition was shown to render the micellization phenomenon more exothermic, the effect being larger as the chain length of alcohol increased . These effects were attributed to the location of alcohol molecules between the surfactant units, their hydroxyl groups close to the surfactant head-groups, in competition with the surfactant counterions . The individual isotherms of alcohol and surfactant adsorption onto XO15M and SiAl32d22 were determined . The plots of the pseudo-differential molar enthalpy of displacement, Delta(dpl)h, against the surface coverage by the surfactant cation, Theta(BDDA+), were derived from the titration calorimetry data . The formation of surface-bound aggregates was thought to be a prerequisite for alcohol coadsorption at the solid-solution interface . At least two different types of adsolubilization sites were postulated, one of the sites being the same as in micelles and the other related to the contact area between the hydrophobic surfactant tails and the equilibrium bulk solution . Coadsorption (adsolubilization) of alcohol molecules at such sites was found to increase the exothermic contribution to the enthalpy of displacement per mole of the BDDA+ adsorbed.

Vet Surg, 2004 Mar-Apr, 33(2), 180 - 6
Intraosseous gentamicin perfusion of the distal metacarpus in standing horses; Mattson S et al.; OBJECTIVE: To report tissue gentamicin concentrations after intraosseous (IO) perfusion in standing horses . STUDY DESIGN: In vivo study . ANIMALS OR SAMPLE POPULATION: Twelve horses . METHODS: Sedated horses had a cannulated cortical bone screw inserted into the dorsolateral aspect of the treated metacarpus and a tourniquet applied proximally . Gentamicin (2.2 mg/kg) diluted in sterile saline solution (0.1 mL/kg) was infused through the screw . Two horses were euthanatized at each time interval: 0, 2, 6, 12, 24, and 36 hours . Synovial fluid and bone samples were collected distal to the screw from both forelimbs . Gentamicin concentrations were measured using fluorescence polarization immunoassay . RESULTS: The highest synovial fluid gentamicin concentrations were 385+/-273 microg/mL (mean+/-SD) in the metacarpophalangeal joint, 225+/-205 microg/mL in the proximal interphalangeal joint, 215+/-205 microg/mL in the distal interphalangeal joint, 382+/-195 microg/mL in the digital flexor tendon sheath, and 206+/-161 microg/mL in the navicular bursa . The highest bone concentrations of gentamicin were 55+/-30 microg/g in the distal metacarpus, 34+/-27 microg/g in the proximal, 16+/-15 microg/g in the middle, and 16+/-2.2 microg/g in the distal phalanges, and 27+/-17 microg/g in the proximal and 24+/-11 microg/g in the distal sesamoid bones . CONCLUSION: Standing IO perfusion of gentamicin resulted in local antibiotic concentrations in the synovial structures and bones of the distal aspect of the limb that exceed the reported minimum inhibitory concentration of pathogens commonly implicated in equine orthopedic infections . CLINICAL RELEVANCE: Standing IO perfusion of gentamicin in the distal aspect of the limb should be considered for treatment of orthopedic infections of this region in horses.

J Periodontol, 2004 Jan, 75(1), 169 - 75
Specific antibiotics in the treatment of periodontitis--a proposed strategy; Beikler T et al.; BACKGROUND: The purpose of the present study was to propose a strategy for the selection of antibiotics that specifically target complexes of periodontal pathogens present in patients with periodontitis . METHODS: Seven hundred seventy-four (774) patients with various forms of periodontitis were included in the study . Subgingival plaque samples were taken from the deepest periodontal pockets in each quadrant using a sterile curet, and pooled . Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Eikenella corrodens, Tannerella forsythensis, Prevotella intermedia, and Prevotella nigrescens were identified by polymerase chain reaction, and the prevalence of combinations of these pathogens was determined . To each pathogen complex (PC), i.e., combination of pathogens, those antibiotics were assigned that were most specific according to the published minimum inhibitory concentration (MIC90) values and the gingival crevicular fluid (GCF) concentrations achievable in vivo . Antibiotic GCF concentrations had to be at least 10 times the MIC90 values, and the narrowest spectrum was selected with respect to the assessed periodontal pathogens . RESULTS: Nine major PCs (each > or = 3% of all patients) were found in 73.4% of all patients, whereas 38 minor PCs (each < 3% of all patients) were distributed in 26.6% of all patients . Ten different antibiotic regimens were found to be specific for the total of 46 PCs; i.e., metronidazole and amoxicillin in 11 PCs (55.0% of all patients), metronidazole and amoxicillin/clavulanic acid or metronidazole and ciprofloxacin in 13 PCs (18.9%), amoxicillin in 4 PCs (8.3%), doxycycline in 2 PCs (6.1%), metronidazole in 8 PCs (4.1%), amoxicillin/clavulanic acid in 3 PCs (2.9%), clindamycin in 2 PCs (1.5%), ciprofloxacin in 2 PCs (0.4%), and tetracycline in 1 PC (0.3%) . CONCLUSION: The results of the study indicate that there are at least 46 different combinations of the assessed periodontal pathogens in subjects with periodontitis, and at least 10 different antibiotic regimens might be required to specifically target the various pathogen complexes.

J Control Release, 2004 Mar 24, 95(3), 521 - 33
Novel mucoadhesive buccal formulation containing metronidazole for the treatment of periodontal disease; Perioli L et al.; Mucoadhesive tablets using different mixture of cellulose and polyacrylic derivatives were prepared in order to obtain new formulations containing metronidazole for periodontal disease treatment . All tablets were characterized by swelling studies, ex vivo and in vivo mucoadhesive time, ex vivo mucoadhesion force, in vitro and in vivo release . The best mucoadhesive performance and the best in vitro drug release profile were achieved by using hydroxyethyl cellulose (HEC) and carbomer 940 2:2 ratio . The chosen tablet, containing 20 mg of metronidazole, performed 12 h drug sustained release with buccal concentrations always higher than its MIC.

Sheng Wu Yi Xue Gong Cheng Xue Za Zhi, 2004 Feb, 21(1), 62 - 5
{Investigation of machinable-infiltrated-ceramic glass infiltrating through the aluminous matrix}; Yang X et al.; This investigation was amied at the infiltrative capability of the machinable-infiltrated-ceramic(MIC) glass and the color of the composite after the MIC glass infiltrated through the aluminous matrix with different packing densities . By heating the components to 1100 degrees C for 2 hours, the MIC glass was made to infiltrate through the aluminous matrix with different packing densities . We measured the infiltrative depth and the color parameter and observed the rupture surface of the composite by means of SEM . There was a linear relation between the square of infiltrative depth and the packing density of aluminous matrix . The minimal depth was 3.092 mm . No relationship was noted between the composite's color coefficient and the packing density of aluminous matrix . In the rupture process of the composite, crack deflexion, crystal evulsion, and rupture through crystal could be observed . This experiment proved that the infiltrative characters of MIC glass meet the clinical requirement, the composite's color is steady and the mechanical intensity is stable.

FEMS Microbiol Lett, 2004 Mar 12, 232(1), 7 - 14
TbtABM, a multidrug efflux pump associated with tributyltin resistance in Pseudomonas stutzeri; Jude F et al.; Tributyltin (TBT) is a toxic agent used in marine antifouling paints . Among the bacterial flora of a polluted harbor, TBT-resistant strains of Pseudomonas stutzeri have been isolated . In the strain 5MP1 (TBT minimum inhibitory concentration (MIC) > or =1000 mg l(-1)), TBT resistance was found to be associated with the presence of the operon tbtABM, homologous to the resistance-nodulation-cell division (RND) efflux pump family, as demonstrated by cloning in Escherichia coli . TbtABM exhibited the greatest homology (60.9-84.9%) with the TtgDEF and SrpABC systems, both involved in aromatic compound tolerance in P . putida . TbtABM conferred multidrug resistance (MDR) including to n-hexane, nalidixic acid, chloramphenicol, and sulfamethoxazole (antibiotic MICsx4 for the E . coli host strain carrying the operon) . By polymerase chain reaction amplification and hybridization experiments, the presence of tbtABM was detected in the TBT-sensitive P . stutzeri 3MP1 (TBT MIC 25 mg l(-1)) . However, the latter strain did not seem to express TbtABM . This is the first description of a MDR efflux pump in P . stutzeri, and of a new kind of substrate, TBT, for the RND family of transporters.

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