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%