J Pharmacol Exp Ther, 2002 Aug, 302(2), 666 - 71 Characterization of methotrexate transport and its drug interactions with human organic anion transporters; Takeda M et al.; Life-threatening drug interactions are known to occur between methotrexate and nonsteroidal anti-inflammatory drugs (NSAIDs), probenecid, and penicillin G . The purpose of this study was to characterize methotrexate transport, as well as to determine the site and the mechanism of drug interactions in the proximal tubule . Mouse proximal tubule cells stably expressing basolateral human organic anion transporters (hOAT1 and hOAT3) and apical hOAT (hOAT4) were established . The K(m) values for hOAT1-, hOAT3-, and hOAT4-mediated methotrexate uptake were 553.8 microM, 21.1 microM, and 17.8 microM, respectively . NSAIDs (salicylate, ibuprofen, ketoprofen, phenylbutazone, piroxicam, and indomethacin), probenecid, and penicillin G dose dependently inhibited methotrexate uptake mediated by hOAT1, hOAT3, and hOAT4 . Kinetic analysis of inhibitory effects of these drugs on hOAT3-mediated methotrexate uptake revealed that these inhibitions were competitive . The K(i) values for the effects of salicylate, phenylbutazone, indomethacin, and probenecid on hOAT3-mediated methotrexate uptake were comparable with therapeutically relevant plasma concentrations of unbound drugs . In addition, in the presence of human serum albumin, the K(i) values were comparable with therapeutically relevant total plasma concentrations of drugs . In conclusion, these results suggest that methotrexate is taken up via hOAT3 and hOAT1 at the basolateral side of the proximal tubule and effluxed or taken up at the apical side via hOAT4 . In addition, hOAT1, hOAT3, and hOAT4 are the sites of drug interactions between methotrexate and NSAIDs, probenecid, and penicillin G . Furthermore, it was predicted that hOAT3 is the site of drug interactions between methotrexate and salicylate, phenylbutazone, indomethacin, and probenecid in vivo. Bull Exp Biol Med, 2002 Apr, 133(4), 334 - 5 Protective effect of copper-rutin complex in animals with experimental epilepsy; Tsaryuk VV et al.; Copper-rutin complex (2 mg/kg) completely eliminated epileptiform potentials induced by a combination of chlorpromazine and microwave radiation 1-2 min postinjection and suppressed convulsive activity provoked by application of penicillin to the sensorimotor cortex. Antimicrob Agents Chemother, 2002 Aug, 46(8), 2716 - 9 Activities of ABT-773 against microaerophilic and fastidious organisms; Finegold SM et al.; ABT-773 was tested against 317 fastidious isolates; it inhibited 99% of organisms at a concentration of 4.0 microg/ml . With ampicillin-sulbactam and levofloxacin, only 2 and 6% of these strains, respectively, were resistant at the breakpoint . With clindamycin, penicillin G, and metronidazole, 22, 26, and 58% of the strains, respectively, were resistant. Clin Experiment Ophthalmol, 2002 Aug, 30(4), 297 - 9 Rapid seroconversion to Treponema pallidum and HIV positivity in a patient with retinal vasculitis; Venkatesh P et al.; A patient with idiopathic retinal vasculitis was found to have rapid conversion of serological tests for Treponema pallidum and HIV . The diagnosis was confirmed by Venereal Disease Research Laboratory (VDRL) testing and dark ground illumination for T . pallidum, and enzyme-linked immunosorbent assay (ELISA) and Western blot assay tests for HIV . Following treatment with intravenous penicillin, the fundus lesions resolved and visual acuity recovered from inaccurate light projection to 6/24 in the right eye and from counting fingers close to face to 2/60 in the left eye . Serological tests for T . pallidum in patients with concurrent HIV infection may be unpredictable . Hence, it is important to repeat these tests even in the early treatment phase of patients with retinal vasculitis who have shown initial seronegativity . This enables earlier diagnosis and initiation of specific treatment. Infection, 2002 Jun, 30(3), 175 - 7 Brachyspira (Serpulina) pilosicoli spirochetemia in an immunocompromised patient; Kanavaki S et al.; The case of an elderly immunocompromised man with non-Hodgkin's lymphoma who presented with fever, abdominal pain and bloody diarrhea is described . Brachyspira pilosicoli was isolated from culture . The patient was treated with penicillin G i.v . and became afebrile . B . pilosicoli is a recently recognized enteric pathogen of humans and animals . Intestinal spirochetosis should be included in the differential diagnosis of any immunocompromised or critically ill patient with dysentery. Infection, 2002 Jun, 30(3), 150 - 6 Effect of dexamethasone and pentoxifylline in combination with amoxicillin in the treatment of penicillin-insensitive pneumococcal pneumonia in guinea pigs; Parra A et al.; A fatal guinea pig model of pneumococcal pneumonia was developed in order to evaluate the efficacy of amoxicillin combined with either pentoxifylline or dexamethasone . Parameters assessed were survival time and lung changes (alterations, bacterial colony counts, inducible nitric oxide synthase {iNOS} and cyclooxygenase-2 {COX-2} protein expression) . Animals receiving pentoxifylline (50 mg/kg) showed higher survival rates than controls (p < 0.05) . Animals which received amoxicillin (50 mg/kg), alone or combined, showed significantly higher survival rates than controls (p < 0.05) . Animals dying in spite of receiving amoxicillin alone or combined had lung colony counts significantly lower than those that did not receive the antibiotic (p < 0.001), but their lungs showed identical changes . The correlation between COX-2 protein expression and mortality was rather high (r = 0.75) . The addition of either dexamethasone or pentoxifylline to amoxicillin improved neither survival rates nor lung pathology when compared with the antibiotic alone. J Pediatr, 1974 Jan, 84(1), 49 - 54 Febrile illness in early infancy associated with ECHO virus infection; Linnemann CC Jr et al.; Twenty-three infants less than six weeks of age with fever of 100.4 degrees F, or higher and no evidence of bacterial infection were seen at the Cincinnati General Hospital from July to December, 1971 . Seventeen of these were admitted to the hospital; 15 were treated with penicillin and kanamycin for possible sepsis . Viral isolation was attempted on 21 of these infants, and ECHO viruses were isolated from 14 (66.7 per cent), compared to three (14.3 per cent) of 21 controls . Eleven of the 14 ECHO viruses isolated were type 4, and the other 3 were types 6, 11, and 25 respectively . Acute and convalescent serum was obtained from 11 of the 21 infants; seven had a fourfold rise in antibody to ECHO virus type 4 . Most of the children had fever with irritability, six of the 23 had a fine maculopapular rash, and three had aseptic meningitis . This study suggests that ECHO viruses may be a significant cause of febrile illness in young infants during the summer and fall and may account for illnesses which lead to hospitalization as possible sepsis. Ann Thorac Surg, 2002 Jul, 74(1), 185 - 90 Surgical considerations for pulmonary actinomycosis; Endo S et al.; BACKGROUND: Diagnosis and treatment of pulmonary actinomycosis is difficult without surgical intervention . METHODS: Thirteen patients (10 men, 3 women; mean age, 62 years) underwent pulmonary resection and were given a pathologic diagnosis of pulmonary actinomycosis at our institution between 1976 and 2001 . To clarify when pulmonary actinomycosis should be suspected in patients and the role of surgical intervention, we reviewed preoperative clinical characteristics, computed tomography findings, surgical indication, operative procedure, postoperative clinical course, and outcome . RESULTS: Ten patients (77%) had poor oral hygiene . Twelve patients (92%) were symptomatic, and 10 patients (77%) had hemoptysis . The mean interval between radiographic identification of the abnormality and surgical intervention was 8 months (interquartile range, 3.25 to 8 months) . Computed tomography findings in all cases included radiologic opacity with air bronchogram or a low attenuation area . Lung cancer was diagnosed initially because of computed tomography findings of spiculation or pleural indentation, and operation was required in 8 patients (62%) . The others were diagnosed with chronic pneumonia, and surgical intervention became necessary because of recurrent hemoptysis or prolonged illness . Six patients underwent lobectomy; the others underwent partial resection or segmentectomy . Neither complication nor recurrence has occurred . CONCLUSIONS: When patients, particularly those with poor oral hygiene, show radiologic opacity with an air bronchogram or low attenuation area on the computed tomography scan, pulmonary actinomycosis should be considered and penicillin should be administered as diagnostic therapy . Surgical intervention may be necessary when frequent hemoptysis has no resolution or lung neoplasm cannot be ruled out. Biotechnol Bioeng, 2002 Aug 5, 79(3), 356 - 61 A two-step, one-pot enzymatic synthesis of cephalexin from D-phenylglycine nitrile; Wegman MA et al.; A cascade of two enzymatic transformations is employed in a one-pot synthesis of cephalexin . The nitrile hydratase (from R . rhodochrous MAWE)-catalyzed hydration of D-phenylglycine nitrile to the corresponding amide was combined with the penicillin G acylase (penicillin amidohydrolase, E.C . 3.5.1.11)-catalyzed acylation of 7-ADCA with the in situ-formed amide to afford a two-step, one-pot synthesis of cephalexin . D-Phenylglycine nitrile appeared to have a remarkable selective inhibitory effect on the penicillin G acylase, resulting in a threefold increase in the synthesis/hydrolysis (S/H) ratio . 1,5-Dihydroxynaphthalene, when added to the reaction mixture, cocrystallized with cephalexin . The resulting low cephalexin concentration prevented its chemical as well as enzymatic degradation; cephalexin was obtained at 79% yield with an S/H ratio of 7.7 . Biotechnol Bioeng, 2002 Jun 5, 78(5), 589 - 93 Penicillin acylase-catalyzed ampicillin synthesis using a pH gradient: a new approach to optimization; Youshko MI et al.; The penicillin acylase-catalyzed synthesis of ampicillin by acyl transfer from D-(-)-phenylglycine amide (D-PGA) to 6-aminopenicillanic acid (6-APA) becomes more effective when a judiciously chosen pH gradient is applied in the course of the process . This reaction concept is based on two experimental observations: 1) The ratio of the initial synthesis and hydrolysis rates (V(S)/V(H)) is pH-dependent and exhibits a maximum at pH 6.5-7.0 for a saturated solution of 6-APA; 2) at a fixed 6-APA concentration below saturation, V(S)/V(H) increases with decreasing pH . Optimum synthetic efficiency could, therefore, be achieved by starting with a concentrated 6-APA solution at pH 7 and gradually decreasing the pH to 6.3 in the course of 6-APA consumption . A conversion of 96% of 6-APA and 71% of D-PGA into ampicillin was accomplished in an optimized procedure, which significantly exceeds the efficiency of enzymatic synthesis performed at a constant pH of either 7.0 or 6.3 . FEMS Microbiol Lett, 2002 Jul 2, 212(2), 229 - 36 Identification and characterization of the Escherichia coli envC gene encoding a periplasmic coiled-coil protein with putative peptidase activity; Hara H et al.; PM61 is a chain-forming envC strain of Escherichia coli with a leaky outer membrane . It was found to have an oversized penicillin-binding protein 3, which was the result of an IS4 insertion in the prc gene . The other properties of PM61 were caused by the envC mutation . We cloned the envC (yibP) gene and identified the mutation site, causing a single residue substitution, H366Y, in the PM61 envC allele . The gene product was predicted to be a periplasmic protein having coiled-coil structure in the N-terminal region and homology to lysostaphin in the C-terminal region . Overexpression of envC inhibited cell growth, and overexpression of the PM61 mutant allele caused cell lysis . Disruption of the chromosomal envC caused the same defects as the envC point mutation, indicating the gene is dispensable for growth but important for normal septation/separation and cell envelope integrity. J Pharm Biomed Anal, 2002 Aug 1, 29(6), 1031 - 43 Kinetic analysis of thermal decomposition for penicillin sodium salts: model-fitting and model-free methods; Rodante F et al.; A kinetic study on decomposition processes of some penicillin salts was carried out . Both isothermal and dynamic thermogravimetric curves were used . As expected by their complex structures, several steps with different energies were involved in decomposition processes . Model-fitting and -free kinetic approaches were applied to nonisothermal and isothermal data . The kinetic triplet (f(alpha),A and E(a)) related to model-fitting method that defines a single step reaction resulted to be at variance with the multi-step nature of salts-decomposition . The model-free approach represented by the isothermal and nonisothermal isoconversional methods, gave different dependencies of the activation energy on the extent of conversion . The complex nature of the multi-step process of the studied compounds was more easily revealed using a broader temperature range in nonisothermal isoconversional method . The failure in the model-fitting method did not allow calculating shelf life and half-life times. Pol Arch Med Wewn, 2002 Feb, 107(2), 161 - 5 {Pulmonary actinomycosis as a complication of foreign body aspiration}; Zielonka TM et al.; This first polish paper presents respiratory actinomycosis as a complication caused by aspiration of a foreign body . Aspiration of a citrus fruit stone occurred as a result of esophagus stenosis and dysphagia caused by a mistake made by a blind person of drinking a caustic substance . Clinical and bronchoscopic features suggested bronchogenic carcinoma but it's not confirmed by two histopathologic examinations of section from pathological bronchial changes . Diagnosis was based on the morphological picture of sections taken during third fibreoptic bronchoscopy and on cytological sputum examination . Removing the aspired foreign body from bronchus and prolonged penicillin treatment resulted in clinical and radiological amelioration. Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai), 2000, 32(2), 163 - 168 Increase in Hydrophobicity of Signal Peptide Enhances Secretion of Penicillin G Acylase; Yang YG et al.; An artificial strong hydrophobic signal peptide (ASP), containing ten leucines in tandem in the hydrophobic core, was utilized to replace the wild type signal peptide (WTSP) of penicillin G acylase (PAC), by the fusion to its 4 pro site . PAC expression plasmids, including pKKpacdeltaSP, pKKpacWTSP, pKKpacASP, pETpacWTSP and pETpacASP, were constructed . The length and the amino- and carboxyl-terminus amino acid composition of ASP and WTSP were kept identical . The activity assay and Western-blotting analysis were used to study the effect of ASP and WTSP on the secretion of PAC in tac, T7 and dissolved-oxygen regulation expression systems, respectively . Lack of signal peptide in pKKpacdeltaSP resulted in the accumulation of 91 kD PAC precursor (without signal peptide, but with the space peptide between alpha-subunit and beta-subunit) in the cytosol, indicating that the secretion of PAC depends on the signal peptide . In BL21(pKKpacASP) cells, the PAC activity and proprecursor (with signal peptide and space peptide) processing capacity were increased by about 54% and 38.5%, respectively, in comparison with BL21(pKKpacWTSP) cells . Compared with BL21(DE3) (pETpacWTSP), however, the PAC activity and proprecursor processing capacity in BL21(DE3) (pETpacASP) were enhanced by about 69% and 43.5%, respectively . The PAC activity expressed from pETpacASP was about 67% more than that from pETpacWTSP in the dissolved-oxygen-regulated expression system GJ100 . Resulting from the strong hydrophobicity of ASP, therefore, the PAC activity and proprecursor processing capacity were increased by about 63% and 41% on average, respectively, in comparison with WTSP . In conclusion, the increase in hydrophobicity of the signal peptide hydrophobic core enhanced the secretion of penicillin G acylase. Drugs, 2002, 62(10), 1447 - 61 Management issues in syphilis; Pao D et al.; Syphilis is a sexually transmitted infection which is systemic from the outset and has increased in incidence worldwide over the last decade . There has been concern as to whether or not co-infection with HIV can modify the clinical presentation of syphilis and, as a genital ulcer disease, it can facilitate the transmission of HIV infection . Diagnosis is based on the microscopic identification of the causative treponeme and serological testing . Recommendations for the treatment of syphilis have been based on expert opinion, case series, some clinical trials and 50 years of clinical experience . Penicillin, given intramuscularly, is the mainstay of treatment and the favoured preparations for early infectious syphilis are benzathine penicillin as a single injection or a course of daily procaine penicillin injections for 10 to 14 days . The duration of treatment is longer for late syphilis . There has been concern that benzathine penicillin may not prevent the development of neurosyphilis but that is a rare outcome with this therapy . The main alternative to penicillin is doxycycline, but the place of azithromycin and ceftriaxone is yet to be established . It is not necessary to carry out examination of the cerebrospinal fluid in patients with early infectious syphilis but it should be performed in those with neurological or ocular signs, psychiatric signs or symptoms, when there is evidence of treatment failure and in those who are co-infected with HIV . Follow-up is an essential part of management and should be particularly assiduous, for at least 24 months, in those co-infected with HIV . Partner notification should be mandatory to try to contain the spread of infection. Biochem Biophys Res Commun, 2002 Jul 5, 295(1), 55 - 61 C-terminus modification of Streptomyces clavuligerus deacetoxycephalosporin C synthase improves catalysis with an expanded substrate specificity; Chin HS et al.; The biosynthesis of cephalosporins is catalyzed by deacetoxycephalosporin C synthase (DAOCS) . Based on computational, biochemical, and structural analyses, it has been proposed that modification of the C-terminus of DAOCS might be a constructive strategy for engineering improvement in enzyme activity . Therefore, five hydrophilic residues namely N301, Y302, N304, R306, and R307 located in proximity to the C-terminus of Streptomyces clavuligerus DAOCS (scDAOCS) were selected and each substituted with a hydrophobic leucine residue . Substitutions at positions 304, 306, and 307 created mutant scDAOCSs with improved efficiencies in penicillin analog conversion up to 397% . And since it has been previously advocated that the C-terminus is crucial for guiding substrate entry, a truncated mutant DAOCS was constructed to assess its involvement . The truncation of the C-terminus at position 310 in the wild-type scDAOCS resulted in reduction of indiscriminate conversion of penicillin analog but this defect was compensated by the replacement of asparagine with leucine at position 304 . (c) 2002 Elsevier Science (USA). Biochem J, 2002 Jul 1, 365(Pt 1), 303 - 9 Role of alphaArg145 and betaArg263 in the active site of penicillin acylase of Escherichia coli; Alkema WB et al.; The active site of penicillin acylase of Escherichia coli contains two conserved arginine residues . The function of these arginines, alphaArg145 and betaArg263, was studied by site-directed mutagenesis and kinetic analysis of the mutant enzymes . The mutants alphaArg145-->Leu (alphaArg145Leu), alphaArg145Cys and alphaArg145Lys were normally processed and exported to the periplasm, whereas expression of the mutants betaArg263Leu, betaArg263Asn and betaArg263Lys yielded large amounts of precursor protein in the periplasm, indicating that betaArg263 is crucial for efficient processing of the enzyme . Either modification of both arginine residues by 2,3-butanedione or replacement by site-directed mutagenesis yielded enzymes with a decreased specificity (kcat/K(m)) for 2-nitro-5-{(phenylacetyl)amino}benzoic acid, indicating that both residues are important in catalysis . Compared with the wild type, the alphaArg145 mutants exhibited a 3-6-fold-increased preference for 6-aminopenicillanic acid as the deacylating nucleophile compared with water . Analysis of the steady-state parameters of these mutants for the hydrolysis of penicillin G and phenylacetamide indicated that destabilization of the Michaelis-Menten complex accounts for the improved activity with beta-lactam substrates . Analysis of pH-activity profiles of wild-type enzyme and the betaArg263Lys mutant showed that betaArg263 has to be positively charged for catalysis, but is not involved in substrate binding . The results provide an insight into the catalytic mechanism of penicillin acylase, in which alphaArg145 is involved in binding of beta-lactam substrates and betaArg263 is important both for stabilizing the transition state in the reaction and for correct processing of the precursor protein. Minerva Pediatr, 2002 Jun, 54(3), 243 - 7 {Loeffler syndrome and Mycoplasma pneumonitis . A case report}; Celandroni A et al.; We describe a clinical case of Loeffler syndrome occurred in a famale aged 13 years . This syndrome is characterized by fleeting pulmonary infiltrates and blood eosinophilia until 70% . Patients usually are only mildly ill or asymptomatic and recovery occurs from few days to some months . Principle causes are: a) mycetes as Aspergillus; b) helmints as Toxocara, Ancylostoma, Trichinella, Ascaris, Strongyloides, Schistosoma; c) chemical agents such as penicillin, para-aminosalycilic acid, hydralazine, nitrofurantoine, chlorpropamide . This girl came to our observation in good general conditions with murmur reduction on the thorax left side, marked peripheral eosinophilia (E 55% of 6100 white blood cell), right pulmonary infiltrate on RX and CT scan . One month before she had fever, treated with amoxycillin and clavulanic acid . Mantoux, Prick tests for main inhalant allergenes, ACE, repeated stools and seric investigations for parasites, mycetes and organisms, were negative except for IgM anti-Myco-plasma antibodies . Broncholavage showed marked eosinophilia . Smear didn't show any blast . The girl recovered in about 40 days (E 4.1% of 8500 WBC, RX negative) . Our hypotesis is a causative role of amoxycillin in inducing the syndrome, even if this is a rare event, with an overlapping of a Mycoplasma infection. Scand J Infect Dis, 2002, 34(4), 257 - 61 Treatment of latent syphilis in HIV-infected patients with 10 d of benzylpenicillin G benethamine: a prospective study in Maputo, Mozambique; Tattevin P et al.; Neurosyphilis has been reported in HIV-infected patients previously treated with penicillin G benzathine, which does not achieve treponemicidal levels in cerebrospinal fluid . Therapy combining benzylpenicillin G and its repository form benzylpenicillin G benethamine could be a potentially effective alternative enhanced regimen for treating latent syphilis in HIV-infected patients because peak serum and cerebrospinal fluid concentrations would be achieved early post-administration by the former molecule and sustained for 24 h due to the prolonged half-life of the latter . In this study, 23 asymptomatic HIV and Treponema pallidum co-infected patients received 10 d of combined therapy (2 M IU intramuscular once daily) and were followed up at 3, 6 and 12 months . None experienced side effects or clinical symptoms . Of the 18 patients who were evaluated 1 y later, 8 (44.4%) exhibited serological treatment failure, defined as a positive serum rapid plasma reagin test . In conclusion, a 10-d regimen combining penicillin G and penicillin G benethamine seems to be of no benefit compared to currently recommended treatment. J Pharm Biomed Anal, 2002 Jul 1, 29(3), 569 - 78 Monitoring of enzymatic hydrolysis of penicillin G by pyrolysis-negative ion mass spectrometry; Ghassempour A et al.; A pyrolysis-negative ion mass spectrometry (Pyr-NIMS) is used for the monitoring of enzymatic hydrolysis of penicillin G (Pen G) to 6-aminopenicillanic acid (6-APA) and phenyl acetic acid (PAA) . The high sensitivity and rapid response time of Pyr-NIMS allow its application to the simultaneously determination of these compounds . The mass to charge (m/z) values of 262, 156 and 135 of Pen G, 6-APA and PAA respectively, are used for the quantitative measurements by selected ion monitoring (SIM) . The limit of detection (LOD), linearity and relative standard deviation (n=5) are 10 ng ml(-1), 100 ng ml(-1)-1000 mg ml(-1) and 1.5%, respectively The results are compared with high performance liquid chromatography (HPLC) . An important advantage of the presented analytical system is the high linearity of signals without preliminary separation and recalibration . The main and interactive effects of pH, temperature and concentration of Pen G for enzymatic hydrolysis of Pen G are studied . Optimize conditions of pH (8), temperature (28 degrees C) and concentration of Pen G (12% w/v) in real samples are obtained. J Inorg Biochem, 2002 Apr 28, 89(3-4), 279 - 92 Organometallic complexes with biological molecules . XVII . Triorganotin(IV) complexes with amoxicillin and ampicillin; Di Stefano R et al.; Novel triorganotin(IV) complexes of two beta-lactamic antibiotics, 6-{D-(-)-beta-amino-p-hydroxyphenyl-acetamido}penicillin (=amoxicillin) and 6-{D-(-)-alpha-aminobenzyl}penicillin (=ampicillin), have been synthesized and investigated both in solid and solution states . The complexes corresponded to the general formula R(3)Sn(IV)antib*H(2)O (R=Me, n-Bu, Ph; antib=amox=amoxicillinate or amp=ampicillinate) . Structural investigations about configuration in the solid state have been carried out by interpreting experimental IR and 119Sn Mossbauer data . In particular, IR results suggested polymeric structures both for R(3)Sn(IV)amox.H(2)O and R(3)Sn(IV)amp*H(2)O . Moreover, both antibiotics appear to behave as monoanionic bidentate ligands coordinating the tin(IV) atom through ester-type carboxylate, as well as through the beta-lactamic carbonyl . Evidence that in none of these compounds water molecules were involved in coordination, was provided by thermogravimetric investigations . On the basis of 119Sn Mossbauer spectroscopy it can be inferred that tin(IV) was pentacoordinate in all of the complexes in the solid state, showing an equatorial R(3)Sn(IV) trigonal bipyramidal (tbp) configuration . The nature of the complexes in solution state was investigated by using 1H and 13C nuclear magnetic resonance (NMR) spectroscopy, while an 119Sn spectrum was obtained for n-Bu(3)Sn(IV)amp*H(2)O . Although 1H- and 13C-NMR measurements suggested that in dimethyl sulfoxide (DMSO)-d(6) solution the polymeric structure collapsed, due to a solvolysis process of the beta-lactamic carbonyl bonding to the organometallic moiety, the complexes have been shown to maintain the same trigonal bipyramidal configuration at tin(IV) atom by the coordination of a DMSO molecule . Cytotoxic activity of these novel semisynthetic antibiotic derivatives has been tested towards spermatocyte chromosomes of the mussel Brachidontes pharaonis (Mollusca: Bivalvia) using two different chromosome-staining techniques such as Giemsa and CMA(3) . The occurrence of typical colchicinized-like (c-like) mitoses on slides obtained from animals exposed to organotin compounds, directly confirmed the high mitotic spindle-inhibiting potency of these chemicals . In addition, by comparative analysis of spermatocyte chromosomes from untreated specimens (negative controls) and specimens treated with the triorganotin(IV) complexes, structural damages such as 'achromatic lesions' and 'chromosome breakages' have been identified. Mediators Inflamm, 2002 Apr, 11(2), 105 - 11 Cytokine secretion by decidual lymphocytes in transient hypertension of pregnancy and pre-eclampsia; Wilczynski JR et al.; BACKGROUND: Transient hypertension (TH) and preeclampsia (PE) are believed to have different pathophysiology . However, 15-25% of pregnant women initially diagnosed as having TH develop PE . To clarify the immuno-pathogenetical connections between the two syndromes, we studied the pattern of T helper cell (Th)1/Th2 cytokine balance disturbances existing inside maternal decidua in normal pregnancy (NP) and pregnancies complicated with TH and PE . METHODS: Third-trimester decidual tissue was obtained by curettage of uterine cavity during elective caesarean sections in NP (n = 11), TH (n = 17) and PE (n = 21) patients . Cell suspensions were prepared by an electromechanical dispersal method and centrifugated using a standard gradient sedimentation technique . Isolated lymphocytes were placed in medium (RPMI 1640, 10% fetal calf serum, L-glutamine, penicillin, streptomycin) and cultured for 72 h with or without mitogen phytohaemaglutinine (PHA) . The enzyme-linked immunosorbent assay method was used for estimation of interleukin (IL)-2, IL-4, IL-6, IL-10, IL-12 and interferon-gamma (IFN-gamma) in culture supernatant . STATISTICAL ANALYSIS: The Kruskal-Wallis and the Mann-Whitney U tests were used (p < 0.05) . RESULTS: Both spontaneous and PHA-stimulated secretion of Th2-type cytokines IL-6 and IL-10 was decreased in PE patients compared with TH and NP patients . The concentration of Th1-type cytokine IFN-gamma was increased in patients suffering both from TH and PE . CONCLUSION: On the base of decidual cytokine secretion, both PE and TH are syndromes of local Th1/Th2 cytokine balance disturbances as compared with NP, and TH seems to be an intermediate step to PE. Folia Microbiol (Praha), 2002, 47(2), 189 - 92 Evaluation of strains derived from Escherichia coli W as hosts for the expression of penicillin G-acylase-encoding gene cloned on the recombinant plasmid pKA18; Grafkova J et al.; The potential for production of penicillin G-acylase (PGA), encoded by the chromosomal gene pgai, of four strains belonging to a genealogical line derived from the strain Escherichia coli W, was evaluated in a medium with and without the inducer phenylacetic acid (PA) . These strains were used as hosts of the recombinant plasmid pKA18, in which the structural gene pgac isolated from the strain RE3, the best host strain of a line giving the highest production, was cloned . The presence of the inducer reduced the copy number of the plasmid in all recombinant strains . Only in recombinant strain RE3 (pKA18) the reduction of the gene dosage of pgac resulted also in the reduction of the amount of PGA synthesized by the cells . The reduced activity of the cells did not result from a segregation of plasmid-free clones . Also the growth rate was decreased by 20 and 40% in the host and recombinant strains, respectively . The host strain RE3 showing the highest production of PGA was also the best host of the recombinant plasmid in terms of the segregational stability and copy number (198 copies per chromosome) . The recombinant strain RE3 (pKA18) also provided the highest production of PGA. J Bacteriol, 2002 Jul, 184(13), 3749 - 52 Penicillin-binding proteins 1a and 1b form independent dimers in Escherichia coli; Charpentier X et al.; We report here that PBP1a can dimerize but does not interact with PBP1b to form PBP1a/PBP1b heterodimers in Escherichia coli . These findings support the idea of a relevant involvement of dimerization of both PBP1a and PBP1b during murein synthesis and suggest the existence of different peptidoglycan synthesis complexes. J Bacteriol, 2002 Jul, 184(13), 3630 - 9 Contribution of membrane-binding and enzymatic domains of penicillin binding protein 5 to maintenance of uniform cellular morphology of Escherichia coli; Nelson DE et al.; Four low-molecular-weight penicillin binding proteins (LMW PBPs) of Escherichia coli are closely related and have similar DD-carboxypeptidase activities (PBPs 4, 5, and 6 and DacD) . However, only one, PBP 5, has a demonstrated physiological function . In its absence, certain mutants of E . coli have altered diameters and lose their uniform outer contour, resulting in morphologically aberrant cells . To determine what differentiates the activities of these LMW PBPs, we constructed fusion proteins combining portions of PBP 5 with fragments of other DD-carboxypeptidases to see which hybrids restored normal morphology to a strain lacking PBP 5 . Functional complementation occurred when truncated PBP 5 was combined with the terminal membrane anchor sequences of PBP 6 or DacD . However, complementation was not restored by the putative carboxy-terminal anchor of PBP 4 or by a transmembrane region of the osmosensor protein ProW, even though these hybrids were membrane bound . Site-directed mutagenesis of the carboxy terminus of PBP 5 indicated that complementation required a generalized amphipathic membrane anchor but that no specific residues in this region seemed to be required . A functional fusion protein was produced by combining the N-terminal enzymatic domain of PBP 5 with the C-terminal beta-sheet domain of PBP 6 . In contrast, the opposite hybrid of PBP 6 to PBP 5 was not functional . The results suggest that the mode of PBP 5 membrane anchoring is important, that the mechanism entails more than a simple mechanical tethering of the enzyme to the outer face of the inner membrane, and that the physiological differences among the LMW PBPs arise from structural differences in the DD-carboxypeptidase enzymatic core. Ann Dermatol Venereol, 2002 Apr, 129(4 Pt 1), 375 - 9 {Hospitalization criteria for erysipelas: prospective study in 145 cases}; Jegou J et al.; INTRODUCTION: Two recent studies conducted in France among general practitioners have shown that they only hospitalized between 20 to 50 percent of patients with erysipelas seen in private practice . We therefore conducted a hospital-based, prospective study designed to determine the hospitalization criteria for erysipelas, since a number of patients are also hospitalized directly through the emergency department . PATIENTS AND METHODS: This prospective, hospital-based study, included patients hospitalized for clinical diagnosis of erysipelas in 9 centres in north-eastern France . Clinical data were recorded using a standard questionnaire filled-in during the first 72 hours of hospitalization . They included: demographic (name and first name initials, age, sex) and clinical (location of erysipelas) characteristics, as well as the origin of the patient (home, emergency department, other department), the reasons for hospitalization (severity of local or systemic signs, suspicion of deep vein thrombosis, location on the face, age over 60 years, associated diseases, absence of improvement after ambulatory treatment, socioeconomic reasons or on principle hospitalization) . Patients hospitalized by (or without) a general practitioner were compared using Chi-2 and Student t tests . RESULTS: One hundred forty-five adults (80 women and 65 men; mean age 61 +/- 20 years) were included in the study: 89 patients (61 p . 100) were hospitalized by a general practitioner whereas 56 (39 percent) went directly to the emergency department . In 128/145 cases (88 p . 100), erysipelas was localized on the lower limbs . The mean number of reasons for hospitalization per patient was of 2.2 +/- 1.2 . The most frequent reason for hospitalization was an associated disease (diabetes, obesity, alcoholism, immunodeficiency), present in 77 cases (53 p . 100) . Patients hospitalized through a general practitioner were older than those hospitalized directly through the emergency department (68 vs 51 years; p<0.001) . Patients hospitalized by a general practitioner more frequently had an erysipelas located on the lower limbs (94 p . 100 vs 79 p . 100; p<0.001) and the mean number of reasons for hospitalization was greater (2.4 vs 1.7; p<0.001), especially suspicion of deep vein thrombosis and elevated mean age . The treatment started during hospitalization was intravenous penicillin G in 67 cases (46 p . 100), oral pristinamycin in 28 cases (19 p . 100) and intravenous or oral amoxicillin in 9 cases (6 p . 100) . COMMENTS: This study demonstrates the existence of two distinct courses of hospitalization for erysipelas . Patients hospitalized by a general practitioner were older and their erysipelas more frequently located on the lower limbs and deep vein thrombosis was suspected . Our study also shows the emergence of a population of patients younger and without medical supervision, for whom the general practitioner is replaced by the emergency department of the local hospital . Nevertheless, the most frequent reason for hospitalization in both groups is the existence of an associated disease, possibly responsible for further complications. Evol Dev, 2002 May-Jun, 4(3), 212 - 22 Ontogenetic torsion in two basal gastropods occurs without shell attachments for larval retractor muscles; Page LR; Results of this study on two species of vetigastropods contradict the long-standing hypothesis, originally proposed by Garstang (1929), that the larval retractor muscles power the morphogenetic movement of ontogenetic torsion in all basal gastropods . In the trochid Calliostoma ligatum and the keyhole limpet Diodora aspera, the main and accessory larval retractor muscles failed to establish attachments onto the protoconch (larval shell) when the antibiotics streptomycin sulfate and penicillin G were added to cultures soon after fertilization . Defects in protoconch mineralization were also observed . Despite these abnormalities, developing larvae of these species accomplished complete or almost complete ontogenetic torsion, a process in which the head and foot rotate by 180 degrees relative to the protoconch and visceral mass . Analysis by using phalloidin-fluorophore conjugate and transmission electron microscopy showed that myofilaments differentiated within myocytes of the larval retractor muscles and adherens-like junctions formed between muscle and mantle epithelial cells in both normal and abnormal larvae . However, in abnormal larvae, apical microvilli of mantle cells that were connected to the base of the larval retractor muscles failed to associate with an extracellular matrix that normally anchors the microvilli to the mineralized protoconch . If morphogenesis among extant, basal gastropods preserves the original developmental alteration that created gastropod torsion, as proposed by Garstang (1929), then the alteration involved something other than the larval retractor muscles . Alternatively, the developmental process of torsion has evolved subsequent to its origin in at least some basal gastropod clades so that the original alteration is no longer preserved in these clades. Pol Merkuriusz Lek, 2002 Mar, 12(69), 194 - 7 {Value of the leukotriene generation test in diagnosis of immediate hypersensitivity to penicillin}; Medrala W et al.; In this study we have decided to explore the diagnostic usefulness of CAST-ELISA test in patients with immediate allergic reactions caused by penicillin . Peripheral blood leukocytes were stimulated by commercially available penicillin allergens--Penicillin G, Benzyl-penicillin--Polylysine and specially performed Penicillin G conjugated to human serum albumin . Test was performed in 33 allergic persons and in 13 control healthy persons well tolerated penicillin antibiotics with negative skin tests results . CAST-ELISA test gave the positive results in 30.3% when unconjugated penicillin was used and in 18.18% with penicillin--polylysine . The best results were obtained when cells were stimulated with penicillin conjugated to human serum albumin--positive results were observed in 47.82% . Accumulate percentage of positive results was 73.91% . In our opinion CAST-ELISA test is reliable method in diagnosis of immediate allergic reactions caused by penicillin and inclusion of penicillin conjugated to human serum albumin seems to be justifiable. Pediatr Neurosurg, 2002 May, 36(5), 236 - 43 Ninth cranial nerve stimulation for epilepsy control . Part 1: efficacy in an animal model; Patwardhan RV et al.; OBJECTIVE: To characterize the effects of stimulation of the nerve of Hering (HN; cranial nerve nine) in controlling seizure activity using a canine model . METHODS: Using penicillin applied topically to a region of the cerebral cortex, 16 seizure-type continuous epileptiform discharges were generated . Ten specimens of HN (five left-sided and five right-sided) were dissected from the cervical region in five dogs and stimulated at varying parameters to determine the effects in controlling epileptiform activity . Electroencephalography (using a multielectrode array), electrocardiography and other vital signs were continuously monitored for side effects . RESULTS: Resolution of continuous epileptiform activity following stimulation was found in 12 of 16 trials (75%); no spontaneous resolution was noted in the absence of stimulation, and stimulation significantly shortened seizure duration (p < 0.05) . Mean epileptiform activity duration was 139 s prior to stimulation (range 1-432 s), with a mean poststimulation delay of 17 s until resolution and a mean interictal time of 399 s . Two specimens became free of seizure activity for the duration of our study (p < 0.001) . No significant side effects (such as the potentially life-threatening cardiac problems seen with right-sided cervical vagal nerve stimulation) were found with stimulation of either left- or right-sided HN . Stimulation of other regional nerves (e.g . twelfth cranial nerve, nerves of the cervical plexus) failed to yield similar control of epileptiform activity . CONCLUSIONS: The results of this pilot study suggest that stimulation of the HN can successfully control focal seizure activity in the majority of cases . Pending further study, stimulation of the HN may have a role in the management of patients suffering from medically and otherwise surgically refractory epilepsy . Pharmacoeconomics, 2002, 20(6), 357 - 66 Sickle cell anaemia: epidemiology and cost of illness; Nietert PJ et al.; The purpose of this paper was to review the research examining the epidemiology of and costs associated with sickle cell anaemia (SCA) . Although there is general acceptance that Black populations are at greatest risk of the disease, estimates of disease incidence and prevalence vary greatly among different Black populations . In addition, the sickle cell haemoglobinopathy poses a health problem to many other ethnic groups, including populations native to Italy, Greece, Turkey, Saudi Arabia, India, Pakistan, Bangladesh, China, and Cyprus . As penicillin prophylaxis has been shown to reduce the risk of sepsis among children with SCA, many governments have established newborn screening programmes to improve the health outcomes for patients with this disease . As a group, patients with SCA incur large numbers of hospital admissions, emergency department visits, and outpatient visits, often at substantial costs, hence, obtaining adequate health insurance is a problem for many patients . A common theme present in studies reviewed in this article is that a small proportion of patients tends to account for a majority of the total healthcare costs . As new diagnostic methods and treatment options become available, balancing costs associated with SCA and quality of healthcare will continue to present challenges to many healthcare providers and insurers. Biotechnol Prog, 2002 May-Jun, 18(3), 629 - 34 Epoxy sepabeads: a novel epoxy support for stabilization of industrial enzymes via very intense multipoint covalent attachment; Mateo C et al.; Sepabeads-EP (a new epoxy support) has been utilized to immobilize-stabilize the enzyme penicillin G acylase (PGA) via multipoint covalent attachment . These supports are very robust and suitable for industrial purposes . Also, the internal geometry of the support is composed by cylindrical pores surrounded by the convex surfaces (this offers a good geometrical congruence for reaction with the enzyme), and it has a very high superficial density of epoxy groups (around 100 micromol/mL) . These features should permit a very intense enzyme-support interaction . However, the final stability of the immobilized enzyme is strictly dependent on the immobilization protocol . By using conventional immobilization protocols (neutral pH values, nonblockage of the support) the stability of the immobilized enzyme was quite similar to that achieved using Eupergit C to immobilize the PGA . However, when using a more sophisticated three-step immobilization/stabilization/blockage procedure, the Sepabeads derivative was hundreds-fold more stable than Eupergit C derivatives . The protocol used was as follows: (i) the enzyme was first covalently immobilized under very mild experimental conditions (e.g., pH 7.0 and 20 degrees C); (ii) the already immobilized enzyme was further incubated under more drastic conditions (higher pH values, long incubation periods, etc.) in order to "facilitate" the formation of new covalent linkages between the immobilized enzyme molecule and the support; (iii) the remaining epoxy groups of the support were blocked with very hydrophilic compounds to stop any additional interaction between the enzyme and the support . This third point was found to be critical for obtaining very stable enzymes: derivatives blocked with mercaptoethanol were much less stable than derivatives blocked with glycine or other amino acids . This was attributed to the better masking of the hydrophobicity of the support by the amino acids (having two charges). Biotechnol Prog, 2002 May-Jun, 18(3), 483 - 8 Penicillin G acylase-fatty lipid biocomposite films show excellent catalytic activity and long term stability/reusability; Phadtare S et al.; The formation of biocomposite films of the pharmaceutically important enzyme penicillin G acylase (PGA) and fatty lipids under enzyme-friendly conditions is described . The approach involves a simple beaker-based diffusion protocol wherein the enzyme diffuses into the lipid film during immersion in the enzyme solution, thereby leading to the formation of a biocomposite film . The incorporation of the enzyme in both cationic as well as anionic lipids suggests the important role of secondary interactions such as hydrophobic and hydrogen bonding in the enzyme immobilization process . The kinetics of formation of the enzyme-lipid biocomposites has been studied by quartz crystal microgravimentry (QCM) measurements . The stability of the enzyme in the lipid matrix was confirmed by Fourier transform infrared spectroscopy (FTIR) and biocatalytic activity measurements . Whereas the biological activity of the lipid-immobilized enzyme was marginally higher than that of the free enzyme, the biocomposite film exhibited increased thermal/temporal stability . Particularly exciting was the observation that the biocomposite films could be reused in biocatalysis reactions without significant loss in activity, which indicates potentially exciting biomedical/industrial application of these films. Eur J Biochem, 2002 Jun, 269(11), 2735 - 9 The role of arginine residues in substrate binding and catalysis by deacetoxycephalosporin C synthase; Lipscomb SJ et al.; Deacetoxycephalosporin C synthase (DAOCS) catalyses the oxidative ring expansion of penicillin N, the committed step in the biosynthesis of cephamycin C by Streptomyces clavuligerus . Site-directed mutagenesis was used to investigate the seven Arg residues for activity (74, 75, 160, 162, 266, 306 and 307), selected on the basis of the DAOCS crystal structure . Greater than 95% of activity was lost upon mutation of Arg-160 and Arg266 to glutamine or other residues . These results are consistent with the proposed roles for these residues in binding the carboxylate linked to the nucleus of penicillin N (Arg160 and Arg162) and the carboxylate of the alpha-aminoadipoyl side-chain (Arg266) . The results for mutation of Arg74 and Arg75 indicate that these residues play a less important role in catalysis/binding . Together with previous work, the mutation results for Arg306 and Arg307 indicate that modification of the C-terminus may be profitable with respect to altering the penicillin side-chain selectivity of DAOCS. Biochim Biophys Acta, 2002 Jun 3, 1597(2), 292 - 300 pH, inhibitor, and substrate specificity studies on Escherichia coli penicillin-binding protein 5; Stefanova ME et al.; The recent structural determination of Escherichia coli penicillin-binding protein 5 (PBP 5) provides the opportunity for detailed structure-function studies of this enzyme . PBP 5 was investigated in terms of its stability, linear reaction kinetics, acyl-donor substrate specificity, inhibition by a number of active site-directed reagents, and pH profile . PBP 5 demonstrated linear reaction kinetics for up to several hours . Dilution of PBP 5 generally resulted in substantial loss of activity, unless BSA or a BSA derivative was added to the diluting buffer . PBP 5 did not demonstrate a significant preference against a simple set of five alpha- and epsilon-substituted L-Lys-D-Ala-D-Ala derivatives, suggesting that PBP 5 lacks specificity for the cross-linked state of cell wall substrates . Among a number of active site-directed reagents, only some thiol-directed reagents gave substantial inhibition . Notably, serine-directed reagents, organic phosphates, and simple boronic acids were ineffective as inhibitors . PBP 5 was stable over the pH range 4.6-12.3, and the k(cat)/K(m) vs . pH profile for activity against Ac(2)-L-Lys-D-Ala-D-Ala was bell-shaped, with pK(a)s at 8.2 and 11.1 . This is the first complete pH profile, including both acidic and basic limbs, for a PBP-catalyzed DD-carboxypeptidase (CPase) reaction . Based on its structure, similarity to Class A beta-lactamases, and results from mutagenesis studies, the acidic and basic limbs of the pH profile of PBP 5 are assigned to Lys-47 and Lys-213, respectively . This assignment supports a role for Lys-47 as the general base for acylation and deacylation reactions. Wiad Lek, 2002, 55(1-2), 81 - 7 {Difficulties with using T lymphocyte culture as a method for diagnosing allergies to benzylpenicillin}; Zdziarski P; The activation of T lymphocytes upon antigen stimulation plays a crucial role in adverse immune responses including drug-specific hypersensitivity reactions . The helpfulness of conventional tritiated thymidine incorporation assay for penicillin allergy diagnostics is limited . Benzylpenicillin, as a reactive compound, constitutes typical example of hapten . Most of research on penicillin hypersensitivity use benzylpenicillin-albumin (BPO-HSA) conjugates . Thus in this study we describe an in vitro proliferation assay with benzylpenicillin or penicillin and autologous serum protein conjugates . Interestingly these conjugates enhanced incorporation of tritiated thymidyne, when benzylpenicillin did not exert an influence on PBMCs proliferation (correlation coefficient r = -0.0119) . This so-called carrier-effect indicates that benzylpenicillin and serum globulin complexes can take part in penicillin allergy (primary immune response) . Optimal secondary response is obtained when the benzylpenicillin bind the same carrier for both primary and secondary immunization . Father-proliferation assay with modification of responses to phytohaemagglutinin by benzylpenicilloilated serum protein results in significant decrease of incorporation of {3H} thymidyne . Otherwise benzylpenicillin did not modify postmitogenic proliferation of PBMCs . Our findings indicate that the use of penicillin and autologous serum protein conjugates is helpful . This study show the manner in which benzylpenicillin forms T-cell epitopes. Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai), 2001, 33(4), 437 - 442 Inhibitive Effect of N-nitro-L-arginine on Hippocampal Neurons Excitation; Chen XC et al.; The inhibitive effect of N-nitro-L-arginine (L-NNA, a nitric oxide synthase inhibitor) on the excitation of cultured rat hippocampal CA1 neurons, stimulated by penicillin G(PG), was investigated . A rapid intracellular NO production induced by PG (4 000 IU/ml) was disclosed when monitored with laser scanningconfocal microscopy (LSCM) . Pretreatment of L-NNA (0--10 &mgr;mol/L) dose-dependently inhibited the NO production, and the intercellular level of glutamate (15 min after PG stimulation) as well . Meanwhile, L-NNA(1 and 10 &mgr;mol/L) also significantly inhibited the immunoreactive methionine enkephalin (ir-M-ENK) level . Furthermore, the immunoreactive dynorphin B (ir-DYN-B) level was increased significantly by L-NNA (10 &mgr;mol/L) . Finally, beta-FNA (100 &mgr;mol/L, an M-ENK receptor inhibitor) facilitated the inhibitive effect of L-NNA on the Glu level, while nor-BIN (100 &mgr;mol/L, a DYN receptor inhibitor) suppressed that effect . In conclusion,L-NNA could inhibit NO production induced by PG (4 000 IU/ml) stimulation, thuslowering the M-ENK level and increasing the DYN-B level, and resulted in a down-regulation of the Glu level and the neuron excitation. Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai), 2001, 33(6), 671 - 676 Mutagenesis of N-terminal Amino Acid Residues in beta-subunit of Glutaryl-7-amino-cephalosporanic Acid Acylase C130; Zhang N et al.; In many GL-7ACA acylases, the first Ser residue at the N-terminal of beta-subunit is the catalytic center . In order to investigate relationship between the N-terminal structure and catalytic activities, peptide replacement and site-directed mutagenesis were performed at the N-terminal of beta-subunit of GL-7ACA acylase C130 . When the N-terminal 8 amino acid residues of C130 were replaced by the corresponding sequence of penicillin acylases PAC and PGA, respectively, the first mutant B8PAC lost the activity of the acylase, and the second mutant B8PGA had lower activity with the K(m) value increasing from 0.44x10(-3)mol.L(-1) to 0.55x10(-3) mol.L(-1), and the k(cat) decreasing from 4.92 s(-1) to 1.64 s(-1) . Although the substitution of Trp (beta4) by Tyr did not change the K(m) value, the k (cat) decreased to 2.29 s(-1) . When the Trp was substitued by Leu, both the K ( m ) and k ( cat ) values decreased . Compared with the wild type, mutations of Ser (beta3) to Met, Ala and Cys caused decrease of K(m) values by 52.27%, 43.18% and 38.64%, respectively . Mutation of Asn (beta2) to Gln caused the K ( m ) value being increased by 5-fold, and k ( cat ) decreased by 10-fold . These results suggested that the N-terminal amino acid residues of beta-subunit in GL-7ACA acylase C130 are important for enzyme function. Lett Appl Microbiol, 2002, 34(6), 413 - 6 Enzymes for animal feeding from Penicillium chrysogenum mycelial wastes from penicillin manufacture; Nuero OM et al.; AIMS: Several enzymes from mycelial wastes of Penicillium chrysogenum fungal cultures from penicillin manufacture were studied as supplements for animal feeding . METHODS AND RESULTS: Proteins were precipitated with tannic acid, acetone, ethanol or polyethyleneglycol 6000 . The levels of the enzyme activities found in the different precipitates were similar . The activities of invertase, beta-1,3-glucanase, lipase and tannase were higher and those of amylase, cellulase, pectinase, protease and xylanase were lower . Precipitation with tannic acid was selected as an easy and simple method . CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: An enzymatic precipitate from these P . chrysogenum wastes, comparable to a commercial enzymatic additive used in animal feeding, was obtained. BMC Genet . 2002 Apr 25;3(1):6. Occurrence of leu+ revertants under starvation cultures in Escherichia coli is growth-dependent; Jin J et al.; BACKGROUND: Many investigations have reported that advantageous mutations occurred more frequently under selective conditions than those under non-selective conditions . This phenomenon is referred to as adaptive mutation . Their characteristics are that adaptive mutations are directed and growth-independent . The idea of directed adaptive mutation had been objected by some reports, however, the idea of growth-independent adaptive mutation has been held till today . RESULTS: In this paper, we have observed that under leucine starvation conditions, leu+ revertants accumulated as a function of time; leu- to leu+ reverse mutation rates and frequencies were higher than those under non starvation conditions; and no divided cells could be monitored by the penicillin method . These results were similar to the time-dependent manner of adaptive mutation from previous reports . However, leucine concentration determinate experiments revealed that certain traces of leucine, which leaked from the E . coli cells, was almost always present in the culture . More numbers of leu+ revertants appeared when the similar cultures were dropped in small areas on the selective plates than when spread on the whole selective plates . These results have shown that mutations under leucine starving conditions are growth-dependent . Fluctuation analysis of leu+ revertants indicated that leu-leu+ mutation occurred spontaneously and randomly . In addition, the spectra of leuB gene in the revertants proved that mutations under selective conditions were not specific or directed . CONCLUSIONS: The above investigations led to the conclusion (1) that the occurrence of leu+ mutations under starvation conditions was growth-dependent . The occurrence mutations was also similar to that under non-starvation conditions (2) . Under starvation conditions the mutation rates were higher, and was not constant during the long process. Biochim Biophys Acta, 2002 Apr 3, 1589(2), 104 - 11 Assessment of the microbody luminal pH in the filamentous fungus Penicillium chrysogenum; van der Lende TR et al.; The enzymes of the penicillin biosynthetic pathway in Penicillium chrysogenum are located in different subcellular compartments . Consequently, penicillin pathway precursors and the biologically active penicillins have to cross one or more membranes . The final enzymatic step that is mediated by acyltransferase takes place in a microbody . The pH of the microbody lumen in penicillin producing cells has been determined with fluorescent probes and mutants of the green fluorescent protein and found to be slightly alkaline. Chemistry, 2002 Apr 15, 8(8), 1879 - 87 Synthesis of nucleopeptides by employing an enzyme-labile urethane protecting group; Jeyaraj DA et al.; Nucleoproteins are naturally occurring biopolymers in which the hydroxy group of a serine, a threonine, or a tyrosine moiety is linked through a phosphodiester group to the 3'- or 5'-end of a nucleic acid . For the study of the biological phenomena in which nucleoproteins are involved, for example, viral replication, nucleopeptides embodying the characteristic linkage between the peptide chain and the oligonucleotide may serve as powerful tools . However, as a result of the multifunctionality and the pronounced acid and base lability of nucleopeptides, their synthesis requires the application of a variety of orthogonally stable blocking groups, which can be removed under the mildest conditions . We have developed a new mild enzymatic deprotection method, that is, the penicillin G acylase-catalyzed hydrolysis of the N-phenylacetoxybenzyloxycarbony (PhAcOZ) group, for the synthesis of nucleopeptides . We demonstrate the wide applicability of this method by coupling the N-terminally deprotected nucleopeptides 31 a-c with PhAcOZ-protected amino acids and subsequent removal of the N-PhAcOZ group from fully protected nucleotetrapeptides 32 a,b with penicillin G acylase . The reaction conditions are very mild (pH 6.8) so that no undesired side reaction such as cleavage of the nucleotide bond or beta-elimination of the nucleotide was observed. Scand J Urol Nephrol, 2002 Feb, 36(1), 87 - 8 Ureteric obstruction caused by pelvic actinomycosis; Ord J et al.; Ureteric obstruction is a well-known complication of actinomycosis, however its management in previous case reports has been very variable and sometimes mutilating . We report a rare case presenting with ischiorectal abscess that was successfully treated by JJ stenting and penicillin. J Pharm Pharmacol, 2002 Apr, 54(4), 499 - 508 The influence of caprate on rectal absorption of phenoxymethylpenicillin: experience from an in-vivo perfusion in humans; Lennernas H et al.; The aim of this in-vivo perfusion study in humans was to investigate the influence of a penetration enhancer, sodium caprate, on the rectal absorption of phenoxymethylpenicillin and antipyrine . Six subjects, 3 male and 3 female, were included in two separate studies using perfusion solution of different pH (T1 and T2, respectively) . Each in-vivo rectal perfusion investigation lasted for 200 min and consisted of two periods of 100 min, the first serving as a control, and sodium caprate being added in the second period in both T1 and T2 . The concentrations of phenoxymethylpenicillin, antipyrine and sodium caprate in the outlet perfusate were assayed by HPLC, as was the plasma concentrations of phenoxymethylpenicillin . At pH 6.0 (0-100 min) the fraction absorbed (f(abs)) and effective permeability (P(eff)) of phenoxymethylpenicillin were 0.3% and 0.06 x 4 cm s(-1), respectively, and remained unaffected by the addition of sodium caprate . When the same subjects were perfused at pH 7.4, the f(abs) and P(eff) of phenoxymethylpenicillin were 2.4% and 0.11 x 10(-4) cm s(-1) (0-100 min), respectively, also remaining unchanged by addition of sodium caprate (100-200 min) . It was possible to determine the plasma AUC of phenoxymethylpenicillin after addition of sodium caprate in three subjects at both pHs; this was in the range of 14.0-62.8 and 56.4-231 (min micromol L(-1)) at pH 6.0 and 7.4, respectively . Interestingly, there was a correlation between P(eff) for sodium caprate and the individual plasma AUC and C(max) of phenoxymethyl-penicillin, which indicates that the permeability of the enhancer in the tissue upon which it should act is crucial for achieving an effect . The f(abs) and the P(eff) of antipyrine were not affected at either pH when sodium caprate was added to the perfusion solution . In conclusion, the plasma pharmacokinetics of phenoxymethylpenicillin suggested a slightly increased rectal absorption at pH 7.4 in subjects where sodium caprate was transported into the rectal tissue . However, the increased P(eff) for phenoxymethylpenicillin wastoo small to detectfrom the outlet perfusate, which suggests that sodium caprate alone has a limited effect on the permeability in-vivo across the rectal epithelium when it is presented in a solution. Appl Biochem Biotechnol, 2002 Mar, 97(3), 181 - 92 Application of spontaneous suction phase-dispersing (SSPD) extractors in the extraction of penicillin G; Chen J et al.; The extraction of penicillin G from an aqueous solution with butyl acetate (BA) and tributyl phosphate (TBP) as extractants was carried out at pH 4 with spontaneous suction phase-dispersing (SSPD) extractors under various operating conditions . Four kinds of SSPD extractors were tested with results compared to those obtained by using an extractor with mechanical stirrers . Rotation speed and different extraction systems were found to influence the penicillin recovery and the stability of emulsion formed during extraction . The percentage of extraction under optimum conditions was 91% without formation of emulsion . The laser particle size measurement instrument combined with SSPD can be used to measure the emulsion droplet size in situ. Appl Biochem Biotechnol, 2002 Mar, 97(3), 165 - 79 Optimization of yield in kinetically controlled synthesis of ampicillin with immobilized penicillin acylase in organic media; Illanes A et al.; Immobilized penicillin acylase is a moderately priced versatile enzyme, that is able to catalyze the synthesis of derived penicillins and cephalosporins from the corresponding beta-lactam nuclei and proper side-chain precursors . Kinetically controlled synthesis is a better strategy when product yield is a key issue . Yield should increase at reduced water activity by depressing the competing hydrolytic reactions in favor of synthesis; therefore, organic cosolvents can be a suitable reaction media for synthesis . Using response surface methodology and product yield as objective function, temperature and pH were optimized in the kinetically controlled synthesis of ampicillin using previously screened cosolvents and reaction conditions . Optimum pH was 6.0 for ethylene glycol (EG) and glycerol (GL) and 6.6 for 1-2 propanediol (PD); optimum temperature was 30 degrees C for GL and for EG and PD was in the lower extreme of the range studied, optimum lying below 26 degrees C . Maximum molar yields predicted by the model were 58,51, and 46% for EG, GL, and PD, respectively, which were experimentally validated . Highest yield in aqueous buffer was always <40% . Molar yields about 60% compare favorably with values reported for the kinetically and thermodynamically controlled synthesis of ampicillin and other derived penicillins. Ann R Coll Surg Engl, 2002 Mar, 84(2), 106 - 8 Splenectomy after paediatric trauma: could more spleens be saved? Godbole P, Stringer MD. AIMS: Guidelines for the emergency management of paediatric splenic trauma became widely available in 1993 . A regional survey was undertaken to assess the application of Advanced Paediatric Life Support guidelines to children who had undergone splenectomy after trauma . PATIENTS AND METHODS: All children who had undergone splenectomy for a ruptured spleen at 8 district hospitals and 2 teaching hospitals in our region between January 1994 and January 1999 were identified from histopathology departmental records . With appropriate permission their case notes were reviewed . RESULTS: Eleven children (9 males) were identified, all from district hospitals, with a median age at presentation of 11.8 years (range, 6.6-16.8 years) . All presented within 6 h of blunt abdominal trauma . Median systolic blood pressure, pulse rate, haemoglobin concentration and injury severity score (ISS) on admission were 115 mmHg (range, 80-140 mmHg), 108 bpm (range, 84-150 bpm), 12.1 g/dl (range, 10.7-12.8 g/dl) and 17 (range, 17-29), respectively . Three children had additional relatively minor injuries . On admission, 3 children received an intravenous fluid bolus of 20 ml/kg and the remainder received only maintenance fluid requirements or less . Five children were investigated by ultrasound imaging, one by double contrast CT scan, and three by laparoscopy . Two children had no imaging studies prior to laparotomy . Laparotomy was performed by a consultant surgeon in 7 cases and by a specialist registrar in 4 cases . Ten children underwent total splenectomy and one child had a partial splenectomy . Median hospital stay was 7 days (range, 5-10 days) . The child with an ISS of 29 developed a persistent pancreatic fistula and subsequently required a distal pancreatectomy . All children received Pneumovax and penicillin prophylaxis . CONCLUSIONS: All children made a full recovery without surgical morbidity . However, none of these cases fulfilled the recommended criteria for laparotomy in children with blunt abdominal trauma and splenectomy may have been avoidable. Intern Med, 2002 Apr, 41(4), 319 - 22 Pulmonary actinomycosis followed by pericarditis and intractable pleuritis; Shinagawa N et al.; A case of pleuropericarditis caused by Actinomyces israelli is described . The patient first underwent left upper lobectomy because of pulmonary actinomycosis . Seven months later, cardiac tamponade developed . Culture of the bloody pericardial effusion resulted in positive growth of Actinomyces israeli . He was successfully treated with penicillin G, ampicillin, and minocyclin . However, right pleural effusion appeared two months later . Cultures of the effusion again yielded positive growth of the same bacteria . However, the strain had gained resistance to any antibiotics that had been effective before . Accordingly, pleurodesis with minocyclin was undertaken, which was fortunately effective for controlling the pleural effusion. J AOAC Int, 2002 Mar-Apr, 85(2), 355 - 64 Parallux beta-lactam: a capillary-based fluorescent immunoassay for the determination of penicillin-G, ampicillin, amoxicillin, cloxacillin, cephapirin, and ceftiofur in bovine milk; Huth SP et al.; An analytical system was developed for detection of antibiotic residues in bovine milk . The method is based on competitive fluorescent immunoassays in glass capillary tubes (U.S . Patent No . 5,624,850) . The system consists of an assay cartridge containing 4 glass capillaries, a reagent tray with 4 wells of dried reagents, and a Parallux processor, which processes the assay, reads fluorescent output, and reports test results . Minimum sensitivity for detection of 6 beta-lactam antibiotics in bovine milk was determined to be penicillin-G, 3.2 ppb; ampicillin, 2.9 ppb; amoxicillin, 3.6 ppb; cloxacillin, 7.4 ppb; cephapirin, 16.3 ppb; and ceftiofur, 33.7 ppb . The assay system was also specific and sensitive for detection of incurred residues at U.S . Food and Drug Administration tolerance levels: penicillin-G, 5 ppb; ampicillin, 10 ppb; amoxicillin, 10 ppb; cloxacillin, 10 ppb; cephapirin, 20 ppb; and ceftiofur, 50 ppb . There was no interference in detection of minimum sensitivity levels of antibiotic by the presence of somatic cells at approximately 1 x 10(6) cells/mL . Milk containing 3 x 10(6) cells/mL bacteria commonly found in mastitic milk also showed no interference when tolerance levels of antibiotic were present . There was no detectable interference on results by a wide variety of non-beta-lactam drugs. An Med Interna, 2002 Feb, 19(2), 89 - 95 {Syphilis: current therapeutic perspectives}; Pestoni Porven C et al.; Despite many years of study questions regarding the appropriate therapy for syphilis still remain . Alternatives to penicillin for the treatment of syphilis continue to be sought . This article examines the studies relating to syphilis treatment published during the last years and also the new treatment guidelines for every stage of this sexually transmitted disease. Przegl Lek, 2001, 58(11), 975 - 8 {Drug allergy among patients suffering from nasal polyps}; Skladzien J et al.; The study was performed in order to evaluate quantity and morphology of nasal polyps in patients with diagnosed drug allergy to aspirin, antibiotics, polocain, procain and other drugs . The study included 60 patients (27 females, 33 males aged between 18-69 years) . Nasal polyp morphology was examined under electron microscope and light microscope . Morphological evaluation of epithelial and subepithelial structures was done under light microscope, taking not of inflammatory infiltration type, basal membrane, as well as degree of intestinal edema . Evaluation of epithelial cell type, their relative proportion, as well as area percentage covered by cilium, microvilli and squamous epithelium was done under electron microscope . Polyps in patients with idiosyncrasy to polish aspirin and sodium metamizole (12 patients) featured increased eosinophilic infiltration, thickened basal membrane, with elevated eosinophil percentage differential count . The epithelial percentage area was covered by squamous cells without villi . Polyps associated with idiosyncrasy to polish aspirin differed morphology compared to polyps originality from patients allergic to other medications . On the other hand polyps from patients allergic to other drugs, such as: penicillin, procainamide were morphologically (electron light microscope, light microscope) similar to those polyps caused by inflammatory factors. Eur J Biochem, 2002 Apr, 269(8), 2093 - 100 The role of hydrophobic active-site residues in substrate specificity and acyl transfer activity of penicillin acylase; Alkema WB et al.; Penicillin acylase of Escherichia coli catalyses the hydrolysis and synthesis of beta-lactam antibiotics . To study the role of hydrophobic residues in these reactions, we have mutated three active-site phenylalanines . Mutation of alphaF146, betaF24 and betaF57 to Tyr, Trp, Ala or Leu yielded mutants that were still capable of hydrolysing the chromogenic substrate 2-nitro-5-{(phenylacetyl)amino}-benzoic acid . Mutations on positions alphaF146 and betaF24 influenced both the hydrolytic and acyl transfer activity . This caused changes in the transferase/hydrolase ratios, ranging from a 40-fold decrease for alphaF146Y and alphaF146W to a threefold increase for alphaF146L and betaF24A, using 6-aminopenicillanic acid as the nucleophile . Further analysis of the betaF24A mutant showed that it had specificity constants (kcat/Km) for p-hydroxyphenylglycine methyl ester and phenylglycine methyl ester that were similar to the wild-type values, whereas the specificity constants for p-hydroxyphenylglycine amide and phenylglycine amide had decreased 10-fold, due to a decreased kcat value . A low amidase activity was also observed for the semisynthetic penicillins amoxicillin and ampicillin and the cephalosporins cefadroxil and cephalexin, for which the kcat values were fivefold to 10-fold lower than the wild-type values . The reduced specificity for the product and the high initial transferase/hydrolase ratio of betaF24A resulted in high yields in acyl transfer reactions. J Gastroenterol Hepatol, 2002 Apr, 17(4), 482 - 7 Update on the management of Helicobacter pylori infection, including drug-resistant organisms; Goh KL; Helicobacter pylori infection has many different clinical outcomes . Not all infected persons need to be treated . Therefore, indications for treatment have to be clear, and several consensus guidelines have been formulated to aid the medical practitioner in this decision-making process . Triple therapy with a proton pump inhibitor (PPI), in combination with amoxicillin and clarithromycin is the established treatment of choice . For patients with penicillin hypersensitivity, metronidazole can be substituted for amoxicillin . Bacterial resistance to antibiotics is a major factor adversely affecting treatment success . Resistance to metronidazole has been reported in up to 80%, and resistance to clarithromycin in 2-10% of strains cultured . Resistance to either one of the antibiotics has been reported to result in a drop in efficacy of up to 50% . Emergence of resistance to both metronidazole and clarithromycin following failed therapy is a cause for concern; this underlines the need to use the best available first-line therapy . To avoid the emergence of resistance to both key antibiotics, the combination of metronidazole and clarithromycin should be avoided where possible . For failed treatment, several strategies can be employed . These include ensuring better compliance with repeat therapy, and maximizing the efficacy of repeat treatment by increasing dosage and duration of treatment, as well as altering the choice of drugs . Quadruple therapy incorporating a bismuth compound with a PPI, tetracycline and metronidazole has been a popular choice as a "rescue" therapy . Ranitidine bismuth citrate has been shown to be able to overcome metronidazole and clarithromycin resistance; it may be a useful compound drug to use in place of a PPI in "rescue" therapies . In the case of persistent treatment failures, it is useful to consider repeating gastroscopy and obtaining tissue for culture, and then prescribe antibiotics according to bacterial susceptibility patterns . It is also important in refractory cases to review the original indication for treatment and determine the importance of the indication . Curr Opin Infect Dis, 2001 Apr, 14(2), 133 - 7 Lyme disease in central Europe; Hercogova J et al.; Lyme borreliosis is a fascinating disease, the aetiopathology of which is not yet completely known . Different subspecies of Borrelia burgdorferi sensu lato are responsible for the variable clinical course of the disease . Some new cutaneous (alopecia) and ocular (photophobia and retinal vasculitis) manisfestations have been described and the largest prospective study on erythema migrans during pregnancy was published during the last year . Optimal therapy of Lyme borreliosis is still lacking, but doxycycline, amoxicillin, penicillin, and ceftriaxone are recommended most frequently. Altern Lab Anim, 2002 Mar-Apr, 30(2), 177 - 98 Selection of test chemicals for the ECVAM international validation study on in vitro embryotoxicity tests . European Centre for the Validation of Alternative Methods; Brown NA; The European Centre for the Validation of Alternative Methods (ECVAM) has sponsored a large international prevalidation and validation study of three embryotoxicity tests, involving embryonic stem cells, limb bud micromass cultures, and post-implantation whole-embryo cultures . The main objective of the study was to assess the performance of these in vitro tests in discriminating between non-embryotoxic, weakly embryotoxic and strongly embryotoxic compounds . An initial part of the study was to select 20 test substances for the formal validation trial, conducted under blind conditions . A database of in vivo and in vitro developmental toxicity test results was complied on 310 chemicals that had been used in previous validation studies, or suggested for such use, or that had good quality "segment II"-type in vivo data, or for which there were human data . From this database, a shortlist of about 30 candidates was constructed . Because the ECVAM study would not include metabolic activation, chemicals known to require activation for their developmental effects were excluded as candidates, although some known stable metabolites were included . Attempts were made: to include substances of diverse mechanism; to avoid overemphasis on pharmaceuticals; to avoid biologically inert substances as non-embryotoxicants; and to make the list different from those used previously . The candidates were of three categories: Class 3, strongly embryotoxic, was defined as developmentally toxic in all species tested, inducing multiple developmental effects, and with a high A/D ratio . Class 1, non-embryotoxic, was defined as not developmentally toxic at maternally toxic exposures, but which may show some minor embryo/fetal toxicity, which cannot be separated from maternal toxicity . Class 2, weakly embryotoxic, were chemicals of intermediate activity . From this candidate list, chemicals of known receptor (androgen, oestrogen, glucocorticoid, aryl hydrocarbon) mechanisms were excluded, on the basis that simple tests for such activity are already available . In addition, chemicals not freely available were excluded, and an emphasis on human data was applied . The final list of 20 chemicals was: Class 3--6-aminonicotinamide, 5-bromo- 2'-deoxyuridine, hydroxyurea, methylmercury chloride, methotrexate, all-trans-retinoic acid; Class 2--boric acid, dimethadione, lithium chloride, methoxyacetic acid, valproic acid (VPA), 2-propyl-4-pentynoic acid (4-yn-VPA), salicylic acid sodium salt; and Class 1--acrylamide, D-(+)-camphor, dimethyl phthalate, diphenhydramine hydrochloride, 2-ethyl-4- methylpentanoic acid (isobutyl-ethyl-VPA), Penicillin G sodium salt, saccharin sodium hydrate. Ned Tijdschr Geneeskd, 2002 Apr 6, 146(14), 668 - 71 {Tetanus in a young unvaccinated girl after a fall in the street}; Dolman KM et al.; A 4-year-old girl developed tetanus after she had fallen on the street a week before . She had never been vaccinated and despite pressure from the family practitioner, the parents refused to allow her to be given human anti-tetanus immunoglobulin as a matter of principle after the wound had been stitched . Seven days later she was admitted to hospital with trismus and risus sardonicus . Upon initial treatment with human anti-tetanus immunoglobulin and penicillin, and subsequently metronidazole, her clinical condition deteriorated with opisthotonus and life-threatening respiratory insufficiency, upon which she was moved to the intensive-care department where she was intubated and mechanically ventilated for two weeks . Finally she made a complete clinical recovery . Thanks to the extensive national vaccination program, tetanus has become a rare disease in the Netherlands . However, the very serious course and possible fatal outcome warrant a keen attitude and adequate treatment. Scand J Immunol, 2002 Apr, 55(4), 390 - 8 Chlamydia pneumoniae inhibits apoptosis in human epithelial and monocyte cell lines; Airenne S et al.; Chlamydia pneumoniae is an obligate intracellular pathogen with a tendency to cause persistent infections that has been associated with many chronic conditions such as asthma and coronary artery disease . However, its immunopathogenic mechanisms are poorly understood . When aiming to study the impact of C . pneumoniae infection on host cell apoptosis, we found that epithelial infected (HL) cells and macrophages (U937-line) were resistant to staurosporine and tumour necrosis factor (TNF)-alpha-induced physiological apoptosis 48, 72 or 120 h post-infection, as determined by flow cytometry, DNA fragmentation assay and fluorescence microscopy . The antiapoptotic influence was observed even at a late stage of the chlamydial life cycle and was dependent on the chlamydial protein synthesis . The mechanisms involved blockage of mitochondrial cytochrome c release and caspase 3 activation . We also found that during a persistent C . pneumoniae infection induced in vitro by penicillin treatment of cell cultures, the inhibition of apoptosis was extended for up to 120 h of follow-up post-infection and was restricted to the cells carrying chlamydial inclusions . Our findings suggest that inhibition of apoptosis may be one of the pathogenetic mechanisms by which C . pneumoniae infection can mediate the development of chronic diseases. Sheng Li Xue Bao, 2000 Apr, 52(2), 131 - 6 {The role of somatostatin in area CA1 of rat hippocampus in experimental epilepy}; Zhao WJ et al.; In area CA1, intrahippocampal microinjection of somatostatin(SS, 0.03 to 0.3 nmol) caused single or a series of spike-sharp waves in the cortex electroencephalogram (EEG), the spectrum average total power of which is increased dose-dependently from 0.006 to 0.15 nmol . Microinjection of SS into the hippocampal CA1 subregion brought out epileptic-like behavioral responses and aggregated kainic acid (KA)-induced epileptiform activity of rats . With extracellular recording technique, the effects of SS on 114 units epileptiform discharge were investigated in penicillin (PEN)-treated 95 rat hippocampal slices . In response to the application of SS (10(-10), 10(-8), 10(-6) mol/L), the burst frequency of area CA1 62/105 (59.05%) recorded unit discharges was increased significantly, while that of 23/105 (21.9%) was decreased . These results suggest that SS in hippocampal CA1 subregion mainly precipitates epileptiform discharge . The addition of anti-somatostatin antibody (1 1000) to the perfusate partly blocked the epileptiform bursts triggered by PEN perfusion, while this antiepileptic effect of SSAB was reversed by application of SS (10(-8) mol/L) . These indicate that endogenous SS may be involved in epileptiform discharge in area CA1 of hippocampus. Antimicrob Agents Chemother, 2002 May, 46(5), 1273 - 80 Effects of amino acid alterations in penicillin-binding proteins (PBPs) 1a, 2b, and 2x on PBP affinities of penicillin, ampicillin, amoxicillin, cefditoren, cefuroxime, cefprozil, and cefaclor in 18 clinical isolates of penicillin-susceptible, -intermediate, and -resistant pneumococci; Nagai K et al.; Amino acid alterations in or flanking conserved motifs making up the active binding sites of penicillin-binding proteins (PBPs) 1a, 2b, and 2x of pneumococci were correlated with changes in affinities of penicillin, ampicillin, amoxicillin, cefditoren, cefuroxime, cefprozil, and cefaclor for these PBPs . Four penicillin-susceptible (PSSP), eight penicillin-intermediate (PISP), and six penicillin-resistant (PRSP) pneumococci were studied by DNA sequencing of the penicillin-binding sites of the pbp1a, -2x, and -2b genes of strains and by determining 50% inhibitory concentrations of the seven agents for PBP1a, -2x, and -2b . Two PSSP strains had alterations in PBP2x (L(546)-->V) (one strain) or PBP2b (T(445)-->A) (one strain) . All eight PISP strains had at least two alterations--T(338)-->P or A or H(394)-->Y in PBP2X and T(445)-->A in BPB2b . All PRSP strains had the same changes seen in PISP strains, as well as T(371)-->A or S substitutions in PBP1a . The two most resistant PRSP strains had a second change in PBP2x (M(339)-->F) in a conserved motif . The affinities of penicillin and ampicillin for all three PBPs were decreased for PRSP and most PISP strains . The affinity of amoxicillin for PBP1a and -2x was decreased only for PRSP . Cefaclor and cefprozil showed decreased affinity of PRSP but not PISP for all three PBPs . Cefuroxime showed decreased affinity of PISP and PRSP for PBP1a and -2x but no change for PBP2b . Cefditoren showed no difference in PBP affinity based on penicillin or cefditoren MICs, indicating a different PBP target for this agent . Overall, the MICs for and PBP affinities of the strains correlated with the changes found in the PBP active binding sites. Ethiop Med J, 1999 Jul, 37(3), 155 - 61 Rheumatic heart disease among school children in Addis Ababa City: awareness and adequacy of its prophylaxis; Oli K et al.; One of the objectives of this large scale cross-sectional study of school children of the Addis Ababa city was to assess the status of rheumatic heart disease (RHD) prophylaxis among rheumatic heart disease patients identified during the survey . Awareness about the presence of the illness in those affected and reasons for poor coverage, when detected, were also assessed . Sixty of the 9388 school children surveyed were found to have rheumatic heart disease . On interviewing parents of the children with rheumatic heart disease, ten acknowledged being informed of their children's cardiac illness . Of these parents, 15% (or 9/60) had some idea that their children had heart disease related to some form of infection . However, only two of the nine (22%) children whose parents had some idea about their disease were on regular monthly benzathine penicillin prophylaxis in the previous 12 months preceding the interview . Three (33%) of the nine children had six or fewer injections in the 12 months preceding the interview . The remaining 4 parents (44%) reported that their children took treatment that included injections only at the time of initial diagnosis several years earlier and had not had any follow up since then . Their reasons for not seeking medical care for their children included lack of information on prophylaxis, inability to pay for the treatment and distance of the health facilities . The lack of awareness and the extremely low rate of regular prophylaxis, therefore, highlight the need for an urgent control programme that takes active case detection, treatment access and health education into consideration. Cent Afr J Med, 2001 Feb, 47(2), 35 - 8 Injection-induced sciatic nerve injury in Nigerian children; Fatunde OJ et al.; OBJECTIVES: A retrospective study of all children with a diagnosis of sciatic nerve injury managed at the University College Hospital, Ibadan, Nigeria over a 12 year period was carried out in order to determine predisposing factors to the nerve injury and highlighting practical preventive measures . DESIGN: The necessary data was collected from the case files of children seen at the hospital with a diagnosis of sciatic nerve injury, from 1988 to 1999 . RESULTS: There were 27 children aged five months to 12 years with a diagnosis of sciatic nerve injury . Twenty (74%) of the children were aged five years or less . While seven patients (26%) presented within two weeks of development of foot drop consequent on intramuscular (i.m.) injection given on the buttock, 20 patients (74%) presented much later . Fever was the most common complaint for which the injection had been given . The identity of the drugs given was not known in 10 patients . In the remaining 17 patients drugs administered were specified and included Chloroquine, Novalgin, Paraldehyde, Procaine penicillin, and Sulfadoxine-Pyrimethamine . Most of the patients had received the injections in privately owned medical facilities where staff with minimal training are often allowed to administer i.m . injections . CONCLUSION: It is suggested that the i.m . route for injection be strongly discouraged when a drug can be given by other routes . Only trained staff should be allowed to administer i.m . injections . Giving i.m . injections at sites other than the buttock maybe advantageous in children particularly those aged five years and below. Sheng Wu Yi Xue Gong Cheng Xue Za Zhi, 2002 Jan, 19(1), 36 - 9 {Investigation on culture of rat pulmonary microvascular endothelial cells and their viscoelasticity}; Li F et al.; It is the infent of this study to establish a simple method for cultivation of rat pulmonary microvascular endothelial cells(PMVECs) and investigate the viscoelasticity of PMVECs . First, we obtained rat's peripheral pulmonary tissue, which then was cut into small pieces and cultured with 3 ml DMEM containing 20% bovine calf serum, 90 U/ml heparin, 4 mmol L-glutamine, 100 U/ml penicillin and 100 micrograms/ml streptomycin . Next, moved away the pulmonary tissue pieces 60 h later, and started passage 2-4 days after continued culture . Last, digested and separated PMVECs and studied viscoelastic coefficients of PMVECs by using micropipette aspiration technique . The results revealed that the cultured PMVECs showed regular cobblestone morphology and conformed with endothelial cells morphological characterization by phase contrast microscopy . PMVECs elastic modulus K1 was 49.3 +/- 9.2 Pa, K2 was 73.2 +/- 24.8 Pa, and it's viscosity factor mu was 19.2 +/- 7.2 Pa . s . These data demonstrate that it is feasible to cultivate PMVECs with tissue pieces method, and PMVECs is of greater rigidity. Biotechnol Bioeng, 2002 May 20, 78(4), 425 - 32 Evaluation of the performance of immobilized penicillin G acylase using active-site titration; Janssen MH et al.; Penicillin G acylase from Escherichia coli was immobilized on Eupergit C with different enzyme loading . The activity of the immobilized preparations was assayed in the hydrolysis of penicillin G and was found to be much lower than would be expected on the basis of the residual enzyme activity in the immobilization supernatant . Active-site titration demonstrated that the immobilized enzyme molecules on average had turnover rates much lower than that of the dissolved enzyme . This was attributed to diffusion limitations of substrate and product inhibition . Indeed, when the immobilized preparations were crushed, the activity increased from 587 U g-1 to up to 974 U g-1 . The immobilized preparations exhibited up to 15% lower turnover rates than the dissolved enzyme in cephalexin synthesis from 7-ADCA and D-(-)-phenylglycine amide . The synthesis over hydrolysis ratios of the immobilized preparations were also much lower than that of the dissolved enzyme . This was partly due to diffusion limitations but also to an intrinsic property of the immobilized enzyme because the synthesis over hydrolysis ratio of the crushed preparations was much lower than that of the dissolved enzyme . Lett Appl Microbiol, 2002, 34(4), 290 - 2 Improvement in the resting-cell bioconversion of penicillin G to deacetoxycephalosporin G by addition of catalase; Gao Q et al.; AIMS: To improve the resting cell bioconversion of penicillin G to deacetoxycephalosporin G (DAOG) by elimination of an oxidizing intermediate which inactivates the enzyme during the reaction . METHODS AND RESULTS: Resting cells of Streptomyces clavuligerus strain NP1 were incubated with penicillin G, required co-factors and decane in the presence of catalase or superoxide dismutase, and production of DAOG was measured . Catalase stimulated the bioconversion but superoxide dismutase did not . CONCLUSIONS: Production of hydrogen peroxide during the ring expansion reaction is at least partially responsible for enzyme inactivation . SIGNIFICANCE AND IMPACT OF THE STUDY: Catalase addition improves the bioconversion and will contribute to the eventual replacement of the current multi-step, expensive and environmentally-unfriendly chemical ring expansion by a biological route. Ann Dermatol Venereol, 2002 Feb, 129(2), 207 - 11 {Cutaneous polyarteritis nodosa in children: three cases}; Assicot C et al.; INTRODUCTION: Polyarteritis nodosa is a necrotizing vasculitis of small and medium-size arteries . The cutaneous form of polyarteritis nodosa follows a chronic course, characterized by recurrent episodes limited to skin, muscles and joints . This entity differs from systemic polyarteritis nodosa in the absence of visceral involvement . This form is rare in children, we describe three cases . CASE REPORTS: We describe three girls with a mean age of 11 years (range: 8-13) . They presented painful subcutaneous edematous nodules, arthralgia and fever . Physical examination revealed livedo reticularis (2 cases) and pharyngeal infection (1 case) . Laboratory findings showed an inflammatory syndrome . Skin biopsy supported diagnosis of polyarteritis nodosa . The course was characterized by periods of remission disrupted by exacerbations, well controlled by salicylotherapy, colchicine, dapsone or penicillin . Corticosteroid therapy was used only for invalidating symptoms . There was no systemic involvement after 2, 5 and 6 years of follow up . DISCUSSION: Cutaneous polyarteritis nodosa in children must be suspected in presence of fever, subcutaneous nodules, livedo reticularis and arthralgia . Prognosis is usually benign, so we recommend no aggressive treatment . In view of the tendency to relapse, long-term follow-up is appropriate, before confirming diagnosis. Parasitol Res, 2002 Feb, 88(2), 172 - 4 An anti-contamination cocktail for the in vitro isolation and cultivation of parasitic protozoa; Maser P et al.; Contamination by bacteria or fungi is a frequent problem with the in vitro isolation and propagation of protozoan parasites . We developed an antibiotic combination of broad anti-contaminant activity but minimal toxicity to protozoa . This anti-contamination cocktail consists of penicillin G (60 microgml(-1)), kanamycin (100 microgml(-1)), chloramphenicol (10 microgml(-1)) and flucytosine (50 microgml(-1)) . It is well tolerated by African and South American trypanosomes, Leishmania spp., Giardia duodenalis, Entamoeba histolytica and Plasmodium falciparum, and it provides an effective means for the treatment of contaminated cultures . The anti-contamination cocktail meets numerous prophylactic applications in which sterile handling is impracticable or impossible . In particular, it facilitates the direct in vitro isolation of parasites from an infected host. Biotechnol Prog, 2002 Mar-Apr, 18(2), 163 - 6 Reciprocal 13C-labeling: a method for investigating the catabolism of cosubstrates; Christensen B et al.; The principle of reciprocal labeling is to use a uniformly 13C-labeled substrate as the primary carbon source and a naturally labeled cosubstrate . Metabolites derived from a naturally labeled cosubstrate, in this case amino acids, can then be identified by their relatively lower content of 13C, and information on the degradation pathway can be deduced . The technique is based on GC-MS measurements of amino acid labeling patterns, making the technique well suited for investigating the relative importance of amino acid biosynthesis and amino acid uptake from the medium, as the 13C content of the amino acids incorporated into biomass is a direct measure of the amino acid biosyntheses . The technique is illustrated by the investigation of the degradation of phenoxyacetic acid, a medium component that is essential for production of penicillin V by Penicillium chrysogenum . Glucose was used as the uniformly labeled primary carbon source. Chembiochem, 2002 Apr 2, 3(4), 332 - 40 Moenomycin-mediated affinity purification of penicillin-binding protein 1b; Stembera K et al.; The antibiotic moenomycin A inhibits the biosynthesis of peptidoglycan, the main structural polymer of the bacterial cell wall . The inhibition is based on a reversible binding of the antibiotic to one of the substrate binding sites at enzymes such as the penicillin binding protein 1b (PBP 1b) . This binding has been employed to isolate PBP 1b by affinity chromatography . Suitable ligands have been prepared from moenomycin A and coupled both to affinity supports and to surface plasmon resonance sensor surfaces . The reactions that take place upon immobilization of the ligands to the affinity support and the sensor surface, respectively, have been studied in detail . With the help of surface plasmon resonance the optimal conditions for binding of PBP 1b to moenomycin-derivated ligands have been established . For the first time the selective binding of the moenomycin sugar moiety to the enzyme has been demonstrated. Psychiatry Clin Neurosci, 2002 Feb, 56(1), 65 - 70 Cerebral blood flow changes in general paresis following penicillin treatment: a longitudinal single photon emission computed tomography study; Kitabayashi Y et al.; Three cases of general paresis were successfully treated with high-dose penicillin . In all cases, cerebrospinal fluid cell counts decreased to normal and mental status improved rapidly . Cerebrospinal fluid protein concentrations decreased and IQ scores and overall levels of functioning improved gradually over 1 year . Regional cerebral blood flow (CBF) changes were analyzed longitudinally for 1 year using single photon emission computed tomography (SPECT) . Autoradiography method and region of interest (ROI) analyses were used for quantitative CBF assessment and 3D stereotactic surface projections (3D-SSP) analyses were used for qualitative CBF pattern assessment . 3D-SSP could not reveal remarkable CBF pattern changes through the courses . The ROI analyses showed remarkable CBF decreases in all brain regions 1 month after the treatment, which recovered to normal levels 1 year after the treatment . These results suggest that remarkable decreases of quantitative CBF counts reflect the disappearance of encephalitis, while their gradual recovery reflects the gradual improvement of cerebral functional activity . As Treponema pallidum infection affects whole brain and CBF changes globally, quantitative CBF assessment may be more efficient than qualitative CBF pattern analyses for the purpose of understanding the pathophysiology of general paresis. Clin Exp Allergy, 2002 Feb, 32(2), 270 - 6 Controlled administration of penicillin to patients with a positive history but negative skin and specific serum IgE tests; Torres MJ et al.; BACKGROUND: Although subjects with a positive history of immediate allergy to penicillin and negative skin test are traditionally considered to tolerate penicillin, current evidence indicates that they may develop an immediate reaction despite negative skin and serum specific IgE tests . It is thought that these patients require additional tests to confirm the diagnosis . OBJECTIVE: To assess in a large group of patients with a history of immediate allergy to penicillins but with both skin test and CAP-FEIA-negative to classical and side chain penicillin determinants, the role of controlled administration of betalactams as a diagnostic test . METHODS: A group of 330 patients with a history of immediate allergic reactions to penicillins was studied by two evaluators from the same allergy unit using the following protocol: skin tests with major and minor determinants of benzylpenicillin (benzylpenicilloyl-poly l-lysine and minor determinant mixture), amoxicillin and ampicillin, and determination of specific IgE antibodies to penicillins, by CAP-FEIA, in serum . If both tests proved negative, a controlled administration of the drug was then carried out . RESULTS: A total of 89 (27%) patients were skin test and CAP-FEIA-negative and therefore required controlled administration of the drug . Of these, 49 developed an immediate response and were therefore considered allergic, and the remainder had good tolerance after administration of both benzylpenicillin and amoxicillin . The clinical characteristics of this group were similar to the other allergic patients who were skin test or CAP-FEIA-positive, except that they were younger (P < 0.01) . Twenty-two (45%) developed a response to benzylpenicillin and 27 (55%) had a selective response to amoxicillin . Although all reactions appeared within 1 h, a positive correlation was found between the dose inducing the response and the time elapsed from drug administration, for both benzylpenicillin and amoxicillin (P < 0.001) . CONCLUSION: These data indicate that an important number of subjects are not correctly identified if only skin tests and/or CAP-FEIA are used and that this is particularly relevant for side chain-specific reactions and younger subjects . This suggests that new diagnostic tests are required so as to limit the use of controlled administration. Scand J Infect Dis, 2002, 34(2), 88 - 92 Nosocomial transmission of syphilis during haemodialysis in a developing country; Saxena AK et al.; Patients with end-stage renal disease (n = 187) secondary to diverse aetiologies who underwent haemodialysis (HD) between November 1996 and November 2000 were routinely screened for syphilis using the rapid plasma reagin (RPR) test and confirmed by means of a microhemagglutination assay for Treponema pallidum . All the confirmed syphilis patients were asymptomatic and were diagnosed serologically . A true seroprevalence of 6.9% (13/187) and a biological false seropositivity of 4.2% (8/187) for syphilis were recorded . Most (11/13) of the true seropositive patients were aged between 51 and 80 y . Whilst 10/13 patients were true syphilis seropositive at the time of first HD, 3/13 patients became true seropositive an average of 12 months (range 10-14 months) after HD . Penicillin treatment was given to all 13 patients simultaneously only after the appearance of 3 new true syphilis seropositive cases . Complete seroreversion was observed in 4/10 patients in the pre-HD true syphilis seropositive group of presumptive transmitters who became RPR-negative, whereas the 3 new true seropositive cases showed a serial 4-fold decline in RPR titres 12 months after penicillin therapy, suggestive of an active disease with adequate therapeutic response . These results clearly indicate that latent syphilis is prevalent in long-term elderly HD patients . The true seroconversion of 3 new patients who had undergone HD for an average of 12 months is indicative of nosocomial transmission and the silently active nature of the disease, which necessitates regular monitoring of syphilis serology among HD patients. ALTEX, 2002, 19(1), 15 - 20 Novel serum replacement based on bovine ocular fluid: a useful tool for cultivation of different animal cells in vitro; Filipic B et al.; Different mammalian cells in culture have individual nutritional requirements, which are mainly fulfilled by the addition of foetal calf serum (FCS) to the basic medium . Collecting FCS is accompanied with severe animal welfare problems as conscient animals usually are bleeded to death by heart-punctuation without anaesthetics . There exists scientific problems too . Due to the batch-to-batch variability and the relatively high price, different types of serum replacements were introduced . Among them bovine colostrum as a serum substitute for the cultivation of hybridoma cells should be mentioned . The presented experiments were aimed to introduce the simple and effective serum replacement (SR) based on the bovine ocular fluid . Throughout the experiments the bovine ocular fluid alone and in the combination with the sheep's defibrinated plasma and human serum albumin was tested for the growth of different cells growing as a monolayer: (a) Cell lines: WISH (human amniotic cell lines) and VERO . (b) Primary culture: chicken embryonic fibroblasts, human bone-marrow fibroblasts . All growth experiments were performed in parallel with the Foetal Calf Serum (FCS) of three different sources . All types of cells were cultivated in Eagle's medium + antibiotics (Penicillin, Streptomycin, Gentamycin) . The most effective was the SR containing approximately 35% of sheep's defibrinated plasma and 1.5% of serum albumin in the bovine ocular fluid . During the experiments 1 and 10% of SR-2.05 or FCS in Eagle's medium were used . After 1, 3 and 6 days of cultivation the cells were counted . The results show that the use of SR-2.05 gives a higher number of cells as compared to most batches of FCS . It is also important that practically no adaptation is needed, meaning that the cells could be grown in Eagle's medium + FCS and in the next passage in Eagle's medium + SR-2.05 and vice versa. Pediatrics . 2002 Apr;109(4):e63. Survival with hypopituitarism from congenital syphilis; Nolt D et al.; Congenital syphilis continues to occur despite the advances in testing of pregnant women in many countries and the availability of penicillin since 1943 . This is a report of a child with multiple systemic manifestations of congenital syphilis . This is one of the few cases of survival with pituitary involvement from congenital syphilis and the first noted case in which diabetes insipidus developed . Institution of routine testing in all countries might have prevented the severe manifestations of syphilis seen in this child. Arch Intern Med, 2002 Apr 8, 162(7), 822 - 6 Lack of penicillin resensitization in patients with a history of penicillin allergy after receiving repeated penicillin courses; Solensky R et al.; BACKGROUND: Up to 10% of the population reports an allergy to penicillin, yet more than 80% of these individuals lack penicillin-specific IgE antibodies . A negative result on a penicillin skin test is highly accurate in identifying who can safely receive the antibiotic at the time of testing . However, its negative predictive value for future courses is unknown because it is uncertain whether patients with a history of penicillin allergy are at risk of becoming resensitized . OBJECTIVE: To determine the rate of penicillin resensitization in adult patients with a history of penicillin allergy after they are challenged with repeated courses of oral penicillin . METHODS: Adult patients with a history of penicillin allergy consistent with an IgE-mediated mechanism were recruited and underwent penicillin skin testing . Those with negative skin test results were challenged with 3 successive 10-day courses of penicillin V potassium (250 mg by mouth 3 times a day), providing their penicillin skin test results remained negative prior to each course . Patients with positive skin test results were not challenged . RESULTS: Of 53 patients with initially negative skin test results, 46 completed the protocol, and each tolerated all 3 courses of penicillin with negative skin test results throughout . No patients had a converted skin test result from negative to a positive, yielding a resensitization rate of 0% (upper 95% confidence interval, 2.1%) . CONCLUSIONS: Adult patients with a history of penicillin allergy are not at increased risk of resensitization after receiving 3 courses of oral penicillin . Because a negative penicillin skin test result is predictive for subsequent oral administrations beyond the time of testing, adult patients with a history of penicillin allergy can be skin tested electively, which may avoid unnecessary treatment with alternate broad-spectrum antibiotics. Przegl Epidemiol, 2001, 55(4), 511 - 5 {Borrelia burgdorferi infection in children}; Duszczyk E et al.; OBJECTIVE: The aim of our analysis of causes of borreliosis in children hospitalized or ambulatory treated in the Clinic of Infectious Disease in Childhood of Medical University in Warsaw . METHODS AND MAIN OBSERVATIONS: We observed 113 children from 6 month to 15 years old . Erythema migrans was diagnosed in 97 cases and in 3 cases was multiple erytheme migrans . RESULTS AND CONCLUSIONS: Neuroborreliosis was diagnosed in 11 children . In the treatment neuroboreliosis the ceftriaxon and cristalline penicillin was administrated. Med J Aust, 2002 Feb 18, 176(4), 176 - 8 Howard Florey, Alexander Fleming and the fairy tale of penicillin; Goldsworthy PD et al.; The public myth of the discovery of penicillin is an archetypal "quest story" of the type common to every human culture . But the real story of the discovery, testing and refinement of penicillin is a complex tale of accident, serendipity, oversight, conflict, the pressure of war, idiosyncratic personalities and even--the invention of history. Crit Care Med, 2002 Jan, 30(1), 77 - 82 Elevation of plasma peptidoglycan and peripheral blood neutrophil activation during hemorrhagic shock: plasma peptidoglycan reflects bacterial translocation and may affect neutrophil activation; Shimizu T et al.; OBJECTIVE: To investigate the relations among bacterial transloation, plasma peptidoglycan elevation, and peripheral blood neutrophil activation during hemorrhagic shock . DESIGN: Prospective, randomized, unblinded animal study . SETTING: Surgical research laboratories of Shiga University of Medical Science . SUBJECTS: Male, specific pathogen-free Sprague-Dawley rats . INTERVENTIONS: The rats were randomly divided into three groups: a conventional group with normal intestinal flora (NF), an antibiotic (streptomycin and penicillin G) decontaminated group (AD), and a sham shock group with normal intestinal flora . The NF and AD groups were subjected to hemorrhagic shock (mean arterial pressure 30 mm Hg, for 30 to 90 mins) . Rats were killed at 30, 60, and 90 mins after shock induction . Systemic blood and mesenteric lymph nodes (MLNs) were cultured for the determination of bacterial translocation (BT) . Systemic plasma peptidoglycan and endotoxin concentrations were measured . To evaluate peripheral blood neutrophil activation, phagocytosis and hydrogen peroxide generation were assayed by flow cytometry . MEASUREMENTS AND MAIN RESULTS: In the NF group, BT to MLNs was significa