Eur J Biochem, 2001 Mar, 268(5), 1298 - 303 Expression of penicillin G acylase from the cloned pac gene of Escherichia coli ATCC11105 . Effects of pacR and temperature; Dai M et al.; The structural gene pac in Eschericia coli ATCC11105 encodes penicillin G acylase (PGA) . Within the pac gene, there is a regulatory gene pacR, which is transcribed in the opposite direction . Site-directed mutagenesis was performed at base 1045 of pac by replacing a T with a C . This substitution did not alter the amino-acid sequence of PGA, but changed the translation start codon of pacR from AUG to GUG . The expression of the mutant pacR decreased dramatically and the lacZ transcriptional fusion analysis showed that GUG was an extremely poor initiation codon for pacR . The pacR mutation caused PGA expression to be constitutive rather than inductive in two strains (E . coli A56, DH10B) . The pac inducer phenylacetic acid (PAA) gave significant induction of PGA production at a concentration of 0.2% in wild type, but PAA at this concentration inhibited both cell growth and PGA production in the pacR mutated strains . The temperature-dependent expression character of pac is preserved in the pacR translation-initiation mutant and the optimum temperature of PGA production was 22 degrees C in both wild type and mutant . At a higher temperature of 37 degrees C, the PGA precursor polypeptide could not be matured into subunits and formed inclusion bodies, as revealed by western blot analysis . Our investigations confirmed the hypothesis of pacR-mediated PAA induction for PGA expression and clarified the inhibitory effect of high temperature upon the post-translational processing of the PGA precursor polypeptide. Fresenius J Anal Chem, 2000 Nov, 368(6), 624 - 6 Rapid liquid chromatographic determination of residual penicillin G in milk; Furusawa N; A simple and rapid high-performance liquid chromatographic (HPLC) method for determination of residual penicillin G (benzylpenicillin, PCG) in milk was developed . The sample preparation was performed by stirring with ethanol and reacting with 5 M 1,2,4-triazole-mercury (II) chloride solution at 65 degrees C for 10 min followed by an ultra centrifugation step . The HPLC separation was carried out using a Mightysil RP-4GP column, a mobile phase of acetonitrile and 0.1 M phosphate buffer (pH 6.5) (35:65, v/v) and a photo-diode array detector . The average recoveries from spiked PCG (0.004, 0.01, 0.05 and 0.1 microgram/mL) were above 86% with coefficients of variation between 1.2 and 4.5% . The limit of detection was 0.004 microgram/mL . This value corresponds to the maximum residue limit (MRL) in milk (0.004 microgram/mL, EU and Japan) . The total time required for the analysis of one sample was below 40 min. Biomol Eng, 2001 Mar, 17(3), 113 - 7 DNA architecture and transcriptional regulation of the Escherichia coli penicillin amidase (pac) gene; Stojcevic N et al.; The transcriptional regulation of Escherichia coli ATCC11105 penicillin amidase (pac) gene was studied by modifying DNA sequences responsible for promoter activation by cyclic AMP receptor protein (CRP) . The nucleotide sequence of the 5'-flanking region of the pac gene contains putative tandem CRP binding sites positioned at -69/-70 and at -111/-112 with respect to the transcriptional start site . Our results obtained with either point mutations or insertion or deletion mutants (each of which rotated the helix structure at the CRP binding site one-half turn) showed significant decrease of penicillin amidase (PA) activity, suggesting the CRP as a major activator . In this study, the evidence for the importance of spacing between tandem binding sites for CRP as well as for their location related to the promoter core sequence has been provided . Involvement of integration host factor (IHF) as an additional regulatory protein in the pac gene transcription regulation was also analyzed . It is shown that activation of the pac gene transcription is elevated by IHF. Nippon Rinsho, 2001 Feb, 59(2), 367 - 73 {Mechanism of drug resistance in Helicobacter pylori}; Maeda S et al.; Clarithromycin is one of the most important antibiotics for H . pylori eradication . However, 5-10% was reported to be resistant . It has been shown that one point mutation in the 23S rRNA gene is associated with resistance to clarithromycin . To detect H . pylori infection and the mutation simultaneously, we have designed PCR primers specific for H . pylori, and established assays of PCR-RFLP and PCR-preferential homo-duplex formation (PHFA) . Using this assay, we can detect mixed infections with wild and mutant-strains . The prevalence of mutant infection increased through clarithromycin-based eradication . However, the existence of mutant strains had been confirmed before therapy in most cases who 'converted' to mutant after therapy . Metronidazole is also one of the most important antibiotics for eradication . However, 5-50% was reported to be resistant . It has been shown that rdx gene mutation is associated with resistance . It is reported that inactivation of the rdx gene is frequently, but not always, associated with resistance to metronidazole . Amoxicillin resistant strains were rare (1.2% in Japanese strains) . It is reported that penicillin-binding protein might play a role in the resistance . By detecting of the resistance based on the molecular mechanism, patients can be treated with adequate antibiotics with information about resistance. Rev Mal Respir, 2000 Dec, 17(6), 1089 - 93 {Prognosis of asthmatic adolescents in adulthood}; Rance F et al.; Little is known about the long-term outcome of asthmatic adolescents . According to the literature, 20 to 50% of asthmatic adolescents become symptom-free when they become adults . We report 50 cases of asthmatic adolescents, 25 girls and 25 boys, with a mean follow of 4.4 years (range 1-12 years) . Only 4 patients (8%) became symptom-free, all 4 were stage I and II patients . One death occurred subsequent to penicillin allergy, but was not related to status asthmaticus . Nor remission was observed in stage III and IV patients, whereas 5.8% and 12% of stage II and II patients respectively grew out of their asthma . The factors predictive of persistent asthma in adulthood were severe asthma at the age of 13 years, female gender, onset of asthma before the age of 7 years, no periodic rhinitis, and exercise induced asthma. J Pediatr Hematol Oncol, 2001 Feb, 23(2), 112 - 6 Parental health beliefs and compliance with prophylactic penicillin administration in children with sickle cell disease; Elliott V et al.; PURPOSE: Prophylactic penicillin is effective in preventing severe invasive pneumococcal infection in children with sickle cell disease . In some families, compliance has been problematic . The aims of this study were to monitor compliance and to assess the efficacy of the Health Belief Model (HBM) in predicting compliance . METHODS: Fifty mothers of children with sickle cell disease, ages 6 to 60 months, participated in the study . On enrollment, mothers completed surveys assessing their health beliefs regarding sickle cell disease and infections . Compliance was assessed through self-reporting by the mothers and through review of local pharmacy records of penicillin refills . RESULTS: Sixty percent of the mothers reported that they were highly compliant with obtaining the prescribed 14-day refills . Pharmacy records indicated that only 12% actually adhered to this schedule . The self-reports were significantly related to compliance ratings; mothers who admitted less than optimal compliance averaged 42 days between refills, compared with 19 days for mothers who reported good compliance . Varying perceptions identified through the HBM accounted for approximately 30% of the variance in compliance rates . The perceived burdens of picking up the refills and remembering to administer the medication were the most significant factors . CONCLUSIONS: Educational efforts alone are not sufficient to ensure compliance with penicillin prophylaxis . Routinely monitoring compliance through pharmacy records, reviewing parental beliefs about sickle cell disease and infections, and exploring barriers to treatment will promote dialogue about the importance of strict compliance with this relatively simple yet life-saving prophylaxis. J Neurosurg, 2001 Feb, 94(2), 270 - 80 Anticonvulsant effects of gamma surgery in a model of chronic spontaneous limbic epilepsy in rats; Chen ZF et al.; OBJECT: The management of intractable epilepsy remains a challenge, despite advances in its surgical and nonsurgical treatment . The identification of low-risk, low-cost therapeutic strategies that lead to improved outcome is therefore an important ongoing goal of basic and clinical research . Single-dose focal ionizing beam radiation delivered at necrosis-inducing and subnecrotic levels was investigated for its effects on seizure activity by using an established model of chronic recurrent spontaneous limbic seizures in rats . METHODS: A single 90-minute period of repetitive electrical stimulation (inducing stimulus) of the hippocampus in rats elicited a single episode of status epilepticus, followed by a 2- to 4-week seizure-free period . Spontaneous recurrent seizures developed subsequently and persisted for the duration of monitoring (2-10 months) . Simultaneous computerized electroencephalography and video recording were used to monitor the animals . After the establishment of spontaneous recurrent seizures, bilateral radiation centered in the ventral hippocampal formation was administered with the Leksell gamma knife, aided by a stereotactic device custom made for small animals . A center dose of 10, 20, or 40 Gy was administered using a 4-mm collimator . Control animals were subjected to the same seizure-inducing stimulus but underwent a sham treatment instead of gamma irradiation . In a second experiment, the authors examined the effects of gamma irradiation on the proclivity of hippocampal neurons to display epileptiform discharges . Naive animals were irradiated with a single 40-Gy dose, as already described . Slices of the hippocampus were prepared from animals killed between 1 and 178 days postirradiation . Sensitivity to penicillin-induced epileptiform spiking was examined in vitro in slices prepared from control and irradiated rat brains . CONCLUSIONS: In the first experiment, single doses of 20 or 40 Gy (but not 10 Gy) reduced substantially, and in some cases eliminated, behaviorally and electrographically recognized seizures . Significant reductions in both the frequency and duration of spontaneous seizures were observed during a follow-up period of up to 10 months postradiation . Histological examination of the targeted region did not reveal signs of necrosis . These findings indicate that single-dose focal ionizing beam irradiation at subnecrotic dosages reduces or eliminates repetitive spontaneous seizures in a rat model of temporal lobe epilepsy . In the second experiment, synaptically driven neuronal firing was shown to be intact in hippocampal neurons subjected to 40-Gy doses . However, the susceptibility to penicillin-induced epileptiform activity was reduced in the brain slices of animals receiving 40-Gy doses, compared with those from control rats that were not irradiated . The results provide rational support for the utility of subnecrotic gamma irradiation as a therapeutic strategy for treating epilepsy . These findings also provide evidence that a functional increase in the seizure threshold of hippocampal neurons contributes to the anticonvulsant influence of subnecrotic gamma irradiation. Afr J Med Med Sci, 1999 Sep-Dec, 28(3-4), 185 - 7 Routine antenatal syphilis screening--a case against; Obisesan KA et al.; To highlight the deficiencies associated with the routine antenatal syphilis screening as it is done now in the University College Hospital, Ibadan, and determine the cost effectiveness or otherwise of syphilis screening using as parameters the specificity of the method of screening as well as the gravity of the disease . The results of VDRL tests performed on pregnant women attending the antenatal clinic of the University College Hospital, Ibadan, in a 10-year period, January 1988 to December 1997 were analysed for seropositivity as well as congenital syphilis . The case notes of 110 sero-reactive patients were retrieved and analysed for pregnancy outcome . The prevalence rate of seropositive patients was 1.1% . Only 3 of the seropositive had repeated tests and were treated empirically with high doses of penicillin . There was no case of congenital syphilis . This study has shown that the syphilis screening as it is done now is not cost-effective . If VDRL test is to be continued, efforts must be made to reintroduce TPHA-test, which is more specific-specificity; it wastes a lot of time and money of the patients . Hence it is not cost effective . If VDRL test is to be continued efforts must be made to reintroduce TPHA-test, which is more specific. Acta Derm Venereol, 2000 Sep-Oct, 80(5), 378 - 80 A two-step schedule for the treatment of actinomycotic mycetomas; Ramam M et al.; Actinomycotic mycetomas usually respond slowly to treatment with antibiotics . In an attempt to hasten clinical resolution, we used a 2-step regimen consisting of an intensive phase of therapy with penicillin, gentamycin and co-trimoxazole for 5-7 weeks, followed by maintenance therapy with amoxicillin and co-trimoxazole . Seven patients were treated, all of whom showed significant reduction in discharge and swelling after the intensive phase . Maintenance therapy was continued for 2-5 months after the lesions became completely inactive . Five patients completed maintenance therapy, which was given for 6-16 months (mean 10.7 months), and remained free of disease during a mean post-treatment follow-up period of 6.4 months . The other 2 patients also responded satisfactorily and continue to receive maintenance therapy . Side-effects necessitating a modification of the treatment schedule occurred in 2 patients but reversed on discontinuation of the drugs responsible . This treatment schedule produces a rapid clinical response during the initial, intensive phase and promotes compliance with the longer maintenance phase of treatment necessary to achieve a complete cure. Intensive Care Med, 2000 Nov, 26(11), 1698 - 700 Critical illness onychomadesis; Wester JP et al.; OBJECTIVE: To present our observation of the development of a rare nail deformity in the prolonged course of disease of a critically ill patient with a pulmonary abscess . DESIGN: Case report . SETTING: Tertiary referral, 16-bed, level I surgical ICU in an academic hospital . PATIENT: A 48-year-old Caucasian male was treated with penicillin for a pneumococcal meningitis and pneumonia . He developed a large pulmonary abscess of the right upper lobe and needed prolonged mechanical ventilation . Extensive surgical treatment was successful eventually . A remarkable feature concerned the occurrence of onycholysis of all finger nails and toe nails resulting in complete shedding of the nails (onychomadesis) . This phenomenon can be regarded as an extreme manifestation of Beau's lines precipitated by a severe systemic insult . CONCLUSION: We observed the development of onychomadesis in a critically ill patient with a large pulmonary abscess . This association has not been described before. Crit Rev Microbiol, 2000, 26(4), 205 - 20 Penicillin binding proteins, beta-lactams, and lactamases: offensives, attacks, and defensive countermeasures; Koch AL; A strong outer covering of peptidoglycan (the sacculus) is essential for most bacteria . Beta-lactams have evolved billions of years ago and can block saccular growth of the organism . This led to the evolution of beta-lactamases and resistant penicillin binding proteins (PBPs) . With the introduction of lactam antibiotics by the pharmaceutical industry, resistance genes in nature were laterally transferred to antibiotic-treated disease-causing organisms and additional modification of beta-lactamase genes and of the regulatory genes of the mecA region took place . However, it can be concluded that very little of either type of resistance mechanisms represents new basic evolution against the penicillin type antibiotics . In the last 60 years the resistant bacteria in the main arose by movement of genes from other organisms, from minor genetic changes, and from alteration of the regulation of synthesis. Biologist (London), 2000 Jun, 47(3), 115 - 9 The discovery of penicillin: so many get it wrong; Diggins FW; Alexander Fleming discovered penicillin in September 1928, the most likely date being Monday 3 September on his return from holiday in Suffolk . This discovery has attracted more myths and misinformation than probably any other in the history of medicine. Rev Fac Cien Med Univ Nac Cordoba, 2000, 57(1), 51 - 7 {Collagenase production increases in rheumatoid arthritis and osteoarthritis synoviocytes incubated}; Montrull HL et al.; Cartilage is a specialized connective tissue . It contains few cells into an extracell matrix . The matrix mainly constituents are collagen and proteoglycans . Its degradation depends on synoviocytes activity, that secrete metalloproteases, agents to proteoglycans catabolism . There are two types of synoviocytes: macrophagics (type "A:') and fibroblastics (type "B") . The proteoglycan destruction can be LT-dependent or LT-independent . The aim of this work is synoviocytes function ex vivo study, free immune system influence . In order to do it, heparinized synovial fluid samples were obtained from 6 osteoarthritic (OA) and 6 arthritic (RA) both sex untreated patients, diagnosed according ACR criteria, which disease duration was longer than 6 months . Patients average age was 70 +/- 2 years . Control samples were synovial fluid from traumatic arthritis or non inflammatory bone-muscle pathology . Synovial fluid was centifugated at 1500 g for 30 minutes to isolate synoviocytes . Sediment containing cells was 6 hs incubed with Dulbecco-Eagles media, that has HEPES Gibco (26 mM); NaHCO3 (0.5 g/I); glutamine (2 mM), streptomicine (100 mg/l), G-penicillin (1 U/ml); anphotericine B (2.5 mg/l) . Cells calification and viability were cytopathologically determined . Before and after incubation, collagenase activity was measured by ELISA-double-sandwich, using 10 micrograms/ml monoclonal anti-MMPs in phosphate-buffer-saline . The antigen-antibody complex production with inespecific proteins was blocked by bovine seric albumine . Streptavidin peroxidase was added and washed with 2,2,azin,di(3-ethyl-benztazoilinsuiphonic) acid to develop color . The link of labeled antibody by absorbance at 410 nm was determined in ELISA-spectrophothometer . RA patients earlier MMPs synoviocytes production was 1373 +/- 115 ng/ml . Then 6 hs incubation 2143 +/- 132 ng/ml was reached . The increase (56%) had high significance (p < 0.0001) . OA earlier MMPs cells production was 276 +/- 23 ng/ml, but after incubation it reached 542 +/- 47 ng/ml . (96% increased with highly significativa difference too: p < 0.0001) . Microscopic study was carried out before and after incubation, and shows a lot of synoviocytes with plenty of cytopiasme when the collagenase leveis were highest . On the contrary, when low MMPs production by synovial fluid, as no incubated osteoarthritic material, a few cells containing picnotics nucleous were observed . Significant quantitative differences in AR and OA enzymatic secretion were observed . Although in rheumatoid arthritic MMPs leveis synoviocytes production were 4.6 times than OA levels, after 6 hs incubation percentage of increase in OA cells secretion was highest . Described results confirm MMP-1 synthesis by synoviocytes, and these levels correlate with inflammation, more pronounced in acute (RA) than chronic pathology (OA) . Synoviocyte incubation let us to test disease changes in synovial fluid according to cells number and phagocytic activity . Authors agree to assert that synovial fluid may reflect what is happening in an articular cartilago, because SF provides markers of joint disease . MMPs are giving information about pathways involved in OA and RA cartilage degradation. Antimicrob Agents Chemother, 2001 Mar, 45(3), 870 - 7 Penicillin-binding proteins in Leptospira interrogans; Brenot A et al.; The Leptospira interrogans ponA and pbpB genes were isolated and characterized . ponA and pbpB encode the penicillin-binding proteins (PBPs) 1 and 3, respectively . There is little sequence variation between the PBP genes from two L . interrogans strains (serovar icterohaemorrhagiae strain Verdun and serovar pomona strain RZ11) . The deduced L . interrogans PBP 1 and PBP 3 protein sequences from the two strains shared over 50% similarity to homologous proteins from Escherichia coli . It was demonstrated for strain Verdun that ponA and pbpB are transcribed individually from their own promoter . The ponA and pbpB genes from both strains are separated by 8 to 10 kb and oriented such that their transcription is convergent . The L . interrogans PBP 1 and PBP 3 proteins were synthesized in E . coli and were modified with ampicillin using a digoxigenin-ampicillin conjugate . These data show that both genes encode functional PBPs. Biotechnol Bioeng, 2001 Feb 20, 72(4), 379 - 88 Quantitative analysis of Penicillium chrysogenum Wis54-1255 transformants overexpressing the penicillin biosynthetic genes; Theilgaard H et al.; The low penicillin-producing, single gene copy strain Wis54-1255 was used to study the effect of overexpressing the penicillin biosynthetic genes in Penicillium chrysogenum . Transformants of Wis54-1255 were obtained with the amdS expression-cassette using the four combinations: pcbAB, pcbC, pcbC-penDE, and pcbAB-pcbC-penDE of the three penicillin biosynthetic genes . Transformants showing an increased penicillin production were investigated during steady-state continuous cultivations with glucose as the growth-limiting substrate . The transformants were characterized with respect to specific penicillin productivity, the activity of the two pathway enzymes delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine synthetase (ACVS) and isopenicillin N synthetase (IPNS) and the intracellular concentration of the metabolites: delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine (ACV), bis-delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine (bisACV), isopenicillin N (IPN), glutathione (GSH), and glutathione disulphide (GSSG) . Transformants with the whole gene cluster amplified showed the largest increase in specific penicillin productivity (r(p))-124% and 176%, respectively, whereas transformation with the pcbC-penDE gene fragment resulted in a decrease in r(p) of 9% relative to Wis54-1255 . A marked increase in r(p) is clearly correlated with a balanced amplification of both the ACVS and IPNS activity or a large amplification of either enzyme activity . The increased capacity of a single enzyme occurs surprisingly only in the transformants where all the three biosynthetic genes are overexpressed but is not found within the group of pcbAB or pcbC transformants . The indication of the pcbAB and pcbC genes being closely regulated in fungi might explain why high-yielding strains of P . chrysogenum have been found to contain amplifications of a large region including the whole penicillin gene cluster and not single gene amplifications . Measurements of the total ACV concentration showed a large span of variability, which reflected the individual status of enzyme overexpression and activity found in each strain . The ratio ACV:bisACV remained constant, also at high ACV concentrations, indicating no limitation in the capacity of the thioredoxin-thioredoxin reductase (TR) system, which is assumed to keep the pathway intermediate LLD-ACV in its reduced state . The total GSH pool was at a constant level of approx . 5.7 mM in all cultivations . Biochemistry (Mosc), 2000 Dec, 65(12), 1367 - 75 Kinetics of ampicillin synthesis catalyzed by penicillin acylase from E . coli in homogeneous and heterogeneous systems . Quantitative characterization of nucleophile reactivity and mathematical modeling of the process; Youshko MI et al.; Kinetic regularities of the enzymatic acyl group transfer reactions have been studied using ampicillin synthesis catalyzed by E . coli penicillin acylase as an example . It was shown that ampicillin synthesis proceeds through the formation of an acylenzyme-nucleophile complex capable of undergoing hydrolysis . The relative nucleophile reactivity of 6-aminopenicillanic acid (6-APA) is a complex parameter dependent on the nucleophile concentration . The kinetic analysis showed that the maximum yield of antibiotic being synthesized depended only on the nucleophile reactivity of 6-APA, the ratio between the enzyme reactivities with respect to the target product and acyl donor, and the initial concentrations of reagents . The parameters characterizing the nucleophile reactivity of 6-APA have been determined . The algorithm of modeling the enzymatic synthesis has been elaborated . The proposed algorithm allows the kinetics of the process not only in homogeneous, but also in heterogeneous ("aqueous solution-precipitate") systems to be quantitatively predicted and described based on experimental values of parameters of the reaction . It was shown that in heterogeneous "aqueous solution-precipitate" systems PA-catalyzed ampicillin synthesis proceeds much more efficiently compared to the homogeneous solution. Catheter Cardiovasc Interv, 2001 Feb, 52(2), 237 - 9 Syphilitic aortitis; Cohen MG et al.; A case of syphilitic aortitis, complicated by bilateral coronary ostial stenosis, in a 40-year-old man is described . Treatment included coronary artery bypass grafting and a drug regimen of penicillin . At 3-month follow-up, an exercise stress test revealed no signs of ischemia. Enzyme Microb Technol, 2001 Feb 1, 28(2-3), 218 - 224 Production of immobilized penicillin acylase using aqueous polymer systems for enzyme purification and in situ immobilization; Guan YH et al.; A novel approach for the isolation and purification of penicillin acylase (PA), which couples aqueous two-phase partitioning and enzyme immobilization has been investigated.A PA yield of 90% was achieved by treating E . coli cells with 4% butyl acetate, freeze-thawing step, and pressure homogenization . PA purification (93% recovery) was achieved by (1) removing cell debris via precipitation with polyethylene glycol (PEG 2000); (2) aqueous two-phase partitioning using a PEG 2000 + phosphate system (87% recovery).An in situ enzyme immobilization approach, using oxirane acrylic or aldehyde-agarose beads dispersed in the PEG-rich phase, was explored for the conversion of penicillin G to 6-aminopenicillanic acid . An appropriate immobilization reaction time was found . The catalytic performance of the enzyme, when immobilized, was found not to be affected by recycling of the phase-forming components. J Nutr, 2001 Feb, 131(2), 255 - 61 Vitamin A supplementation at birth delays pneumococcal colonization in South Indian infants; Coles CL et al.; Nasopharyngeal colonization is a risk factor for pneumococcal disease, a leading cause of complications and death in infants . We assessed the impact of vitamin A supplementation in reducing pneumococcal colonization in infants from an area with endemic vitamin A deficiency . We recruited 464 2-mo-old infants from a rural area in South India . Infants were randomly assigned to receive two 7000-microg retinol equivalent doses of vitamin A (n = 239) or placebo (n = 225) orally at birth, and nasopharyngeal specimens were collected at ages 2, 4 and 6 mo . We studied the effect of vitamin A on culture-confirmed pneumococcal colonization and on the distribution of pneumococcal serotypes . Analyses were conducted by intention-to-treat . The risk of colonization among infants aged 4 mo who were not colonized by age 2 mo was significantly reduced in the vitamin A group compared with the placebo group {odds ratio 0.51 (0.28, 0.92), P = 0.02) . The odds of colonization were 27% lower in the treatment group than in the placebo group {odds ratio 0.73 (0.48, 1.1), P = 0.13} . No differences were detected in the prevalence of invasive serotypes . The risk of colonization with penicillin-resistant isolates was 74% lower in the vitamin A group than in the placebo group at 2 mo of age . However, the prevalence of penicillin-resistant isolates was only 4% . Neonatal vitamin A supplementation may play a role in lowering morbidity rates associated with pneumococcal disease by delaying the age at which colonization occurs. Antimicrob Agents Chemother, 2001 Feb, 45(2), 616 - 20 Nucleotide sequence and characterization of a novel cefotaxime-hydrolyzing beta-lactamase (CTX-M-10) isolated in Spain; Oliver A et al.; A cefotaxime-resistant, ceftazidime-susceptible Escherichia coli isolate was obtained from a patient with sepsis in 1997, from which a beta-lactamase with a pI of 8.1 was cloned . Cephaloridine and cefotaxime relative hydrolysis rates were 167 and 81, respectively (penicillin G rate = 100), whereas ceftazidime hydrolysis was not detected . The nucleotide sequence revealed a bla gene related to that coding for CTX-M-3 . Despite 21 nucleotide substitutions, only 2 determined amino acid changes (Ala27Val and Arg38Gln) . The amino acid sequence identity between this enzyme, designated CTX-M-10, and the chromosomal beta-lactamase of Kluyvera ascorbata was 81%. Microbiology, 2001 Feb, 147(Pt 2), 325 - 35 Construction and analysis of ss-lactamase-inhibitory protein (BLIP) non-producer mutants of Streptomyces clavuligerus; Thai W et al.; The gene encoding BLIP, a beta-lactamase-inhibitory protein, was disrupted in wild-type Streptomyces clavuligerus and in a clavulanic acid non-producing mutant . The resulting BLIP mutant and BLIP/clavulanic acid double mutant showed no residual proteinaceous beta-lactamase-inhibitory activity, indicating that only a single beta-lactamase-inhibitory protein exists in S . clavuligerus . The lack of any proteinaceous beta-lactamase-inhibitory activity in the bli and bli/claR mutants also indicates that BLP, the BLIP-like protein, encoded by S . clavuligerus does not possess beta-lactamase-inhibitory activity despite its similarity to BLIP . The bli mutant and the bli/claR double mutant did not show any aberrant growth morphology, sporulation defects, or alterations in cephamycin C production or penicillin G resistance when compared to wild-type S . clavuligerus or to the claR single mutant . Mutants bearing the bli gene disruption did show an elevated level of production of clavam-2-carboxylate and hydroxymethyl clavam as well as clavulanic acid . This phenomenon was observed in the middle stages of production of these clavams but was not detected during maximum production . The production of BLIP was also determined to be down-regulated in a ccaR mutant, lacking the pathway-specific transcriptional regulator required for production of cephamycin C and clavulanic acid . Sequencing of the regions flanking the bli gene showed the presence of a partial open reading frame that encodes a DNA-binding protein, and several open reading frames apparently involved in the production of an ABC transporter. Usp Fiziol Nauk, 2000 Jan-Mar, 31(1), 71 - 80 {The experimental development of the concept of O . S . Adrianov on the correlation of functional and neurochemical processes: regulatory peptides in mediator system dysfunction}; Popova NS et al.; The article is devoted to commemoration of full member of Russian Academy of Medical Sciences, Oleg Andreevich Adrianov, who would have celebrated his 75-th anniversary in 1998 . O . S . Adrianov, author of numerous works on physiology and morphology of central nervous system, in the recent years of his was studying the problem of the processes relationship at macro and micro levels of brain organization . Further to the concept created by O.S . Adrianov, data on action of two peptides: delta-sleep and tafcine, on behavior, neurophysiological and neurochemical processes have been consolidated . Experimental data were obtained for rabbits, cats, and dogs, both intact and in the state of pathology (psychomotoric excitement, bradykinesia, penicillin epilepsy) . Impact of peptides on convergation processes is discussed: peptide of delta-sleep depresses reactions of brain structures to photo- and phono-stimulation, and activates the serotoninergic system in general; tafcine enforces the convergation processes and activates the dopaminergic system. Arch Intern Med, 2001 Jan 8, 161(1), 15 - 21 Anaphylaxis in the United States: an investigation into its epidemiology; Neugut AI et al.; BACKGROUND: Anaphylaxis is a severe, life-threatening allergic reaction that affects both children and adults in the United States . However, data regarding the incidence and prevalence of anaphylaxis and the number of deaths caused by it are limited . OBJECTIVE: To provide a better understanding of the magnitude of the problem of anaphylaxis in the United States . METHODS: A thorough review of the current medical literature was conducted to obtain prevalence estimates on each of the 4 major subtypes of anaphylaxis (food, drugs, latex, and insect stings) . We calculated an overall estimate of the risk of anaphylaxis by using only estimates that are specifically derived from epidemiologic studies measuring anaphylaxis in the general population . RESULTS: Known rates or cases of anaphylaxis were 0.0004% for food, 0.7% to 10% for penicillin, 0 . 22% to 1% for radiocontrast media, and 0.5% to 5% after insect stings . There were 220 cases after latex exposure . Considering the 1999 US population of 272 million, the population at risk for anaphylaxis from food is 1099, from penicillin is 1.9 million to 27 . 2 million, from radiocontrast media is 22 000 to 100 000, from latex is 220, and from insect stings is 1.36 million to 13.6 million . These calculations yield a total of 3.29 million to 40.9 million individuals at risk of anaphylaxis . CONCLUSION: The occurrence of anaphylaxis in the US is not as rare as is generally believed . On the basis of our figures, the problem of anaphylaxis may, in fact, affect 1.21% to 15.04% of the US population. Am J Surg, 2000 Nov, 180(5), 345 - 52 The last fifty years of neonatal surgical management; Rowe MI et al.; Neonatal surgical mortality has steadily fallen over the last five decades . Improved survival does not appear to be related to the introduction of new operative procedures . Most of the basic procedures were developed by 1960 . Eight developments appear to be responsible: (1) The growth of pediatric surgery resulted in widespread availability of neonatal surgeons and dissemination of knowledge about newborn surgical emergencies . (2) The parallel growth of pediatric anesthesia, beginning in 1946, provided specialized intraoperative management of the neonate . (3) Understanding neonatal physiology is the key to successful management; major advances occurred between 1950 and 1970 . (4) New inventions revolutionized patient care; the transistor (1947) made it possible for medical devices to sense, amplify and control physiologic responses and opened the communication and computer age . (5) Neonatal mechanical ventilation had a prohibitive mortality and was seldom utilized; the development of CPAP and a continuous flow ventilator in the 1970s allowed safe ventilatory support . (6) Total parenteral nutrition (1968) prevented starvation that frequently affected infants with major anomalies . (7) The effective treatment of infection began with the clinical use of penicillin (1941); antibiotics have reduced mortality but infants suffering from the septic syndrome have a prohibitive mortality; cytokine, proinflammatory agent research, and the development of anti-inflammatory and blocking agents in the 1980s have not affected mortality . (8) The establishment of newborn intensive care units (1960) provided an environment, equipment, and staff for effective physiologic management. Microsc Res Tech, 2001 Jan 1, 52(1), 137 - 52 Porcine Choroid plexus epithelial cells in culture: regulation of barrier properties and transport processes; Haselbach M et al.; The epithelial cells of the choroid plexus are the structural basis of the blood-cerebrospinal fluid (CSF)-barrier . Here we summarise our recent efforts to culture those cells mainly on permeable supports in vitro . Isolated from porcine brains, we report a simple protocol for the primary culture using cytosine arabinoside as an additive that is cytotoxic for other cells except the plexus epithelial cells . Enhanced barrier properties are obtained by withdrawal of serum from the culture medium after confluency is reached . Cells improve their polarity, permeability for hydrophilic substrates is lowered, electrical resistance is increased tenfold, and a pH-gradient is built up across the cell monolayer . Polarised secretion of proteins and most importantly fluid secretion into the apical filter compartment was attained and proven to be dependent on the Na(+),K(+)-ATPase activity . Active transport processes (penicillin G, riboflavin, myo-inositol, ascorbic acid) were studied and clearly showed the involvement of the organic anion transporter . The permeability of the barrier was found to be regulated by cyclic adenosine monophosphate (cAMP) . Moreover, we report that cell proliferation and differentiation is controlled by components of the extracellular matrix . The present culture model could now be used as an in vitro system to quantify drug transport across the blood-CSF-barrier . J Bacteriol, 2001 Jan, 183(2), 628 - 36 pbpB, a gene coding for a putative penicillin-binding protein, is required for aerobic nitrogen fixation in the cyanobacterium Anabaena sp . strain PCC7120; Lazaro S et al.; Transposon mutagenesis of Anabaena sp . strain PCC7120 led to the isolation of a mutant strain, SNa1, which is unable to fix nitrogen aerobically but is perfectly able to grow with combined nitrogen (i . e., nitrate) . Reconstruction of the transposon mutation of SNa1 in the wild-type strain reproduced the phenotype of the original mutant . The transposon had inserted within an open reading frame whose translation product shows significant homology with a family of proteins known as high-molecular-weight penicillin-binding proteins (PBPs), which are involved in the synthesis of the peptidoglycan layer of the cell wall . A sequence similarity search allowed us to identify at least 12 putative PBPs in the recently sequenced Anabaena sp . strain PCC7120 genome, which we have named and organized according to predicted molecular size and the Escherichia coli nomenclature for PBPs; based on this nomenclature, we have denoted the gene interrupted in SNal as pbpB and its product as PBP2 . The wild-type form of pbpB on a shuttle vector successfully complemented the mutation in SNa1 . In vivo expression studies indicated that PBP2 is probably present when both sources of nitrogen, nitrate and N(2), are used . When nitrate is used, the function of PBP2 either is dispensable or may be substituted by other PBPs; however, under nitrogen deprivation, where the differentiation of the heterocyst takes place, the role of PBP2 in the formation and/or maintenance of the peptidoglycan layer is essential. Rev Mal Respir, 2000 Nov, 17(5), 969 - 72 {Acute pleuro-pneumonitis resulting from leptospirosis}; Mennecier B et al.; We report a case of acute pneumonitis with pleural effusion and respiratory distress syndrome that was the inaugural sign of leptospirosis in a 37-year-old patient exposed to rat dejections at home . The patient was given penicillin and oxygen therapy with evacuation of the pleural effusion . Lung manifestations in leptospirosis usually occur as non-specific cough and hemoptysis . Pleural effusion is uncommon . Adult respiratory distress syndrome and profuse hemoptysis can also occur, requiring special care. Vet Res, 2000 Nov-Dec, 31(6), 623 - 34 Inflammatory response to intramuscular implantation of polyacrylonitrile ultrafiltration probes in sheep; Imsilp K et al.; Polyacrylonitrile is used in the manufacture of dialysis membranes . These membranes are fundamental to the functioning of implantable probes for microdialysis and ultrafiltration sampling of tissue fluids . Although in vivo experimentation using polyacrylonitrile has been reported to cause little inflammatory response when implanted subcutaneously, such information is not available for intramuscular implantation in sheep . The procaine and benzathine salts of penicillin are formulated for intramuscular injection . These salts of penicillin or the formulation excipients may cause inflammatory reactions . Use of polyacrylonitrile probes to draw samples from sites at which these formulations have been injected may be compromised by inflammation or direct interaction between formulation excipients and the dialysis membrane . The aim of this project was to describe tissue responses to intramuscular implantation of polyacrylonitrile in the presence and absence of either procaine or procaine plus benzathine salts of penicillin G . Each of 20 normal sheep was implanted with two ultrafiltration probes, one at the site of an injection of procaine or benzathine plus procaine penicillin G . Similar injections were also made at remote intramuscular sites . After 8, 9, and 11 days of the experiment, sheep were killed and the injection and implantation site muscle were excised and prepared for histopathological examination . The implantation of the probe alone caused greater inflammatory response than the injection of procaine or procaine plus benzathine penicillin G at remote intramuscular sites . The histopathological lesions were greatest where the implantation site was coupled with the injection of either formulation of penicillin G . Polyacrylonitrile may not be a suitable dialysis membrane material for intramuscular implantation in sheep. Bioelectrochemistry, 2000 Dec, 52(2), 229 - 38 Agents facilitating the electric field-induced fusion of intact rabbit erythrocytes; Haritou M et al.; In the recent years, the role of specific membrane active agents in the electrofusion process has started to draw attention, and it has been found that the presence of various substances in the cell medium can affect the fusion process either in a positive or negative way . In this work, the effect of several proteins, bivalent cations and antibiotics was tested with respect to their ability to protect intact erythrocytes from hemolysis and facilitate the fusion process . The effect of different sugars was also studied . Among the different proteins, pronase and proteinase were found to be the most effective . With respect to bivalent cations, Ca2+ and Mn2+ were more effective while Mg2+ was less important . From the antibiotics, penicillin caused a negative effect while streptomycin acted positively . Finally, glucose medium was found to be the most effective compared to all sugars tested . The results indicated that there are strong differentiations of the induced effects caused by each substance, and some possible mechanisms of action of these agents on the erythrocyte membrane were discussed. Eur J Dermatol, 2000 Dec, 10(8), 611 - 3 Linear atrophoderma of Moulin; Rompel R et al.; We report a typical case of linear atrophoderma of Moulin that represents a distinct clinical entity . A 17-year-old woman presented with hyperpigmented and atrophic band-like skin lesions measuring 3-5 cm in breadth on the right side of her trunk and on the right buttock, in an arrangement following the system of Blaschko's lines . The skin lesions had a normal texture and showed no signs of inflammation, lilac ring, erythema, induration, sclerosis or depigmentation . Routine laboratory data were normal . Antinuclear antibodies and anti-Scl70-antibodies were negative . Histopathologically, a moderate diffuse hyperpigmentation within the lower epidermis, a focal vacuolar degeneration of the basal layer, and a few dermal perivascular lymphocytes accompanied by signs of pigment incontinence were noted . Intravenous penicillin G was administered for 14 days in a dosage of 10 x 10(6) IU twice daily . This treatment was repeated after 3 and 9 months . No effect was noted . Linear atrophoderma of Moulin takes a chronic course without progression or regression . The disease leads to significant cosmetic impairment and this may cause emotional stress . Due to its relationship to idiopathic atrophoderma of Pierini-Pasini and linear scleroderma, treatment with intravenous penicillin was tried . In our case it had no effect. Trop Med Int Health, 2000 Nov, 5(11), 800 - 4 Impact on perinatal mortality of missed opportunities to treat maternal syphilis in rural South Africa: baseline results from a clinic randomized controlled trial; Rotchford K et al.; OBJECTIVE: To demonstrate the impact on perinatal mortality of inadequate treatment for maternal syphilis despite adequate screening . METHOD: In 12 clinics providing antenatal care in Hlabisa, South Africa 1783 pregnant women were screened for syphilis at their first antenatal visit between June and October 1998 . Pregnancy outcome was determined among those with syphilis . RESULTS: A total of 158 women were diagnosed with syphilis: prevalence 9% (95% CI 8-10%) . Mean gestation at first antenatal visit was 24 weeks . Thirty women (19%) received no treatment and 96 (61%) received all three recommended doses of penicillin . Among those receiving at least one dose, mean delay to the first dose was 20 days . Among those fully treated mean delay to treatment completion was 34 days . Pregnancy outcome was known for 142 women (90%) and there were 17 perinatal deaths among 15 women (11%) . Eleven of 43 women (26%) who received one or fewer doses of penicillin experienced a perinatal death whilst only four of 99 women (4%) who received two or more doses of penicillin did so (P = 0.0001) . Protection from perinatal death increased with the number of doses of penicillin: linear modelling suggests that one dose reduced the risk by 41%, two doses by 65% and three doses by 79%, compared with no doses . A dose-specific, categorical model confirmed reduction in risk by 79% for all three doses . CONCLUSION: Despite effective screening, many pregnant women with syphilis remain inadequately treated, resulting in avoidable perinatal mortality . Delays in starting and finishing treatment, as well as incomplete treatment occur . Near-patient syphilis testing in the antenatal clinic with early treatment could improve treatment of syphilis and reduce perinatal mortality, and a randomized trial to test this is underway. Enzyme Microb Technol, 2000 Dec, 27(10), 766 - 773 Genetic strategies to enhance penicillin acylase production in Escherichia coli; Chou CP et al.; We demonstrated the improvement of penicillin acylase (PAC) production by optimization of the host/vector system using genetic engineering strategies . Several expression plasmids with improved efficiency for the transcription of the pac gene and/or translation of the pac mRNA were constructed . Mutant strains, isolated by a novel screening method, were effective for use as the expression host to produce PAC . The feasibility of using the mutant strains harboring a selection of expression plasmids for the production of PAC was evaluated . The effect of the mutation(s) resulting in the improved PAC producing ability was characterized . While the production of PAC was significantly enhanced using the optimized host/vector system, the formation of PAC inclusion bodies was shown to be another step limiting the production of recombinant PAC. Hautarzt, 2000 Nov, 51(11), 869 - 71 {Recurrent mitigated erysipelas in severe combined immunodeficiency}; Stark R et al.; A patient with congenital severe combined swiss-type immune deficiency suffered from recurrent cellulitis and at least 26 documented cases of bacterial pneumonia . Parenteral immunoglobins prevented the recurrence of the bacterial pneumonias but not of the cellulitis . This was successfully halted by benzathine penicillin injections every four weeks. Hautarzt, 2000 Nov, 51(11), 838 - 45 {Penicillin allergy as a diagnostic problem . Overview and personal studies}; Walker T et al.; BACKGROUND AND OBJECTIVE: Penicillin allergy is a common clinical problem . The distinction between penicillin and para-infectious exanthems is difficult . We investigated the reliability of the history, as well as the sensitivity and specificity of skin tests and specific IgE levels . PATIENTS/METHODS: 160 patients with a history of penicillin allergy were retrospectively evaluated in the outpatient department of a dermatological clinic . RESULTS: Nearly 50% were diagnosed as allergic to penicillin by detection of specific IgE or skin test . About 60% of the patients with immediate type reactions, and 72% with maculo-papular erythema showed positive reactions in skin tests . Significantly more patients were diagnosed as allergic to penicillin by intradermal testing than by prick testing (p < 0.05) . The sensitivity of the specific IgE RAST was 17.9%; the specifity, 89.5% . For the prick test the sensitivity was 8.2%; the specificity 90.8% . For the intradermal test the sensitivity was 26%; the specifity 69.7% . CONCLUSIONS: We suggest a step by step procedure to detect penicillin allergy making the diagnostic results as valid as possible. Biotechnol Appl Biochem, 2000 Dec, 32 ( Pt 3), 173 - 7 Optimization of 6-aminopenicillanic acid (6-APA) production by using a new immobilized penicillin acylase; Torres-Bacete J et al.; A new immobilized penicillin acylase (ECPVA) was obtained by covalent binding of penicillin acylase from Streptomyces lavendulae on Eupergit C . Enzymic hydrolysis of penicillin V catalysed by ECPVA was optimized using a 2(3) factorial design of experiments, and the selected parameters for this study were pH, temperature and substrate concentration . The immobilized enzyme showed an optimal pH value of 9.5-10.5, and an optimal temperature of 60 degrees C, whereas its soluble counterpart showed the same optimal pH value and a lower optimal temperature of 50 degrees C. J Bacteriol, 2001 Jan, 183(1), 200 - 6 Differential responses of Escherichia coli cells expressing cytoplasmic domain mutants of penicillin-binding protein 1b after impairment of penicillin-binding proteins 1a and 3; Chalut C et al.; Penicillin-binding protein 1b (PBP1b) is the major high-molecular-weight PBP in Escherichia coli . Although it is coded by a single gene, it is usually found as a mixture of three isoforms which vary with regard to the length of their N-terminal cytoplasmic tail . We show here that although the cytoplasmic tail seems to play no role in the dimerization of PBP1b, as was originally suspected, only the full-length protein is able to protect the cells against lysis when both PBP1a and PBP3 are inhibited by antibiotics . This suggests a specific role for the full-length PBP1b in the multienzyme peptidoglycan-synthesizing complex that cannot be fulfilled by either PBP1a or the shorter PBP1b proteins . Moreover, we have shown by alanine-stretch-scanning mutagenesis that (i) residues R(11) to G(13) are major determinants for correct translocation and folding of PBP1b and that (ii) the specific interactions involving the full-length PBP1b can be ascribed to the first six residues at the N-terminal end of the cytoplasmic domain . These results are discussed in terms of the interactions with other components of the murein-synthesizing complex. Biotechnol Bioeng, 2001 Jan 20, 72(2), 185 - 93 Energetics of growth and penicillin production in a high-producing strain of Penicillium chrysogenum; vanGulik WM et al.; The results of a large number of carbon-limited chemostat cultures of Penicillium chrysogenum carried out on glucose, ethanol, and acetate as the growth limiting substrate have been used to obtain an estimation of the adenosine triphosphate (ATP) costs for mycelium growth, penicillin production, and maintenance and the overall stoichiometry of oxidative phosphorylation of the fungus . It was found that penicillin production was accompanied by a significant additional energy drain (73 mol of ATP per mole of penicillin-G) from primary metabolism . This finding has been confirmed in independent experiments and has been shown to result in a significantly lower estimate for the maximum theoretical yield of penicillin-G on the carbon source. Anal Biochem, 2000 Dec 15, 287(2), 196 - 202 Fluorescent coupled enzyme assays for D-alanine: application to penicillin-binding protein and vancomycin activity assays; Gutheil WG et al.; D-Alanine (D-Ala) is a ubiquitous constituent of bacterial cell walls . Assays for D-Ala can be used to investigate several aspects of cell wall biosynthesis and the effects of antibiotics on this process . High-sensitivity fluorescent assays for D-Ala were developed in a microtiter plate format based on d-aminoacid oxidase/horseradish peroxidase (DAO/HRP)-coupled reactions . For comparative purposes the classic chromogenic (UV-vis) assay using o-phenylenediamine (OPD) was also adapted to microtiter plates . OPD gave a lower limit of sensitivity of 2 nmol and was linear up to 60 nmol . Two commercially available fluorogenic HRP substrates were then tested in this assay . Amplex Red (AR) gave a lower limit of sensitivity of 2 pmol and was linear up to 400 pmol d-Ala . QuantaBlu (QB) based assays exhibited a lag in their response to D-Ala corresponding to 50 pmol D-Ala . This lag complicated calibration, but could be eliminated by addition of 150 pmol D-Ala to all assays . The QB assays were linear up to 3000 pmol D-Ala and gave a lower limit of sensitivity of 10 pmol . These assays are demonstrated for the characterization of the dd-carboxypeptidase activity of a soluble form of Escherichia coli penicillin-binding protein 5 (PBP 5) against the classic PBP substrate diacetyl-L-Lys-D-Ala-D-Ala . AR and QB based assays gave identical v/E(T) profiles, whereas OPD based assays gave slightly (10%) higher activity . This is consistent with the loss of a small amount of E . coli PBP 5 activity during the dilution necessary prior to its use in the highly sensitive fluorescent assays . These assays were then demonstrated for characterization of vancomycin binding to a D-Ala-D-Ala-based substrate . Tidsskr Nor Laegeforen, 2000 Oct 30, 120(26), 3128 - 32 {Legionella pneumonia--important differential diagnosis in pneumonia after travelling abroad}; Berdal JE; BACKGROUND: Legionella pneumophila is a rare cause of pneumonia in Norway . From 1992 to 1999, only 27 cases were reported to the Norwegian Surveillance System for Communicable Diseases . MATERIAL AND METHODS: Five cases diagnosed at the Akershus Central Hospital over the last three years are presented . RESULTS: All patients acquired their infection while travelling abroad, mainly in the Mediterranean area, and all fell ill within fourteen days of returning home . The course was serious with marked hypoxaemia in three of the patients, and one patient died . Confusion and altered mental state were prominent features of the clinical presentation . INTERPRETATION: Patients returning from visits abroad and presenting with a pneumonia within to weeks after arriving in Norway, should be investigated for Legionella as a causative agent unless there is a prompt response to empiric penicillin therapy. J Neuroradiol, 2000 Sep, 27(3), 207 - 10 {Syphilitic spinal cord gumma}; El Quessar A et al.; Syphilitic spinal cord gumma Syphilitic gummas of the central nervous system are exceptional and are in general described in the brain . We report the case of a Brown-Sequard syndrome in a 25-year-old patient . The cervical myelography and the brain CT as well as the vertebro-occipital junction were normal . The diagnosis of multiple sclerosis was evoked and corticosteroid therapy was initiated . The patient experienced clinical improvement . Two years later, the patient was readmitted . Immunological reactions for syphilis were positive, in serum and CSF . Tests for HIV were negative . The MRI showed a cervical spinal cord process at the C3 level with adhesive spinal associated arachnoiditis . Penicillin therapy was started prior to surgery for the spinal process . The syphilitic nature was confirmed by pathology . To our knowledge, the MR appearance of a syphilitic gumma of the spinal cord has not been described previously in the scientific literature. Genetics, 2000 Dec, 156(4), 1483 - 92 Selected amplification of the cell division genes ftsQ-ftsA-ftsZ in Escherichia coli; Vinella D et al.; Rapidly growing Escherichia coli is unable to divide in the presence of the antibiotic mecillinam, whose direct target is penicillin-binding protein 2 (PBP2), responsible for the elongation of the cylindrical portion of the cell wall . Division can be restored in the absence of PBP2 activity by increasing the concentration of the cell division proteins FtsQ, FtsA, and FtsZ . We tried to identify regulators of the ftsQ-ftsA-ftsZ operon among mecillinam-resistant mutants, which include strains overexpressing these genes . By insertional mutagenesis with mini-Tn10 elements, we selected for insertions that conferred mecillinam resistance . Among 15 such mutants, 7 suppressed the thermosensitivity of the ftsZ84(Ts) mutant, strongly suggesting that they had increased FtsZ activity . In all 7 cases, however, the mutants resulted from a duplication of the ftsQAZ region . These duplications seemed to result from multiple events, suggesting that no simple insertional inactivation can result in a mutant with sufficiently amplified ftsQAZ expression to confer mecillinam resistance . The structure of the duplications suggests a general method for constructing directed duplications of precise sequences. Ann Allergy Asthma Immunol, 2000 Nov, 85(5), 363 - 5 Frequency of systematic reactions to penicillin skin tests; Valyasevi MA et al.; BACKGROUND: Penicillin skin testing is generally considered to be safe when performed sequentially with puncture and intradermal testing although fatalities have been reported . OBJECTIVE: We analyzed the rate of systemic reactions to penicillin skin tests for a period of seven and two-thirds years . METHOD: This retrospective study used a computerized database at the Mayo Clinic . Altogether 1710 patients were skin-tested to penicillin from January 1992 to September 1999 . All patients tested had a history of penicillin allergy . Patients were tested with benzylpenicilloyl polylysine (Pre-Pen) (6.0 X 10(-5) M), freshly prepared penicillin G (10,000 units/ml), and penicilloate (0.01 M) . Prick tests were done first and if negative then intradermal tests . Systemic reactions were evaluated and treated by physicians . RESULTS: Eighty-six patients had positive penicillin skin tests of which two had systemic reactions . Our systemic reaction rate for all patients tested was 0.12%; and 2.3% for the penicillin skin test-positive group, with no fatalities . CONCLUSION: The incidence of systemic reaction to penicillin skin tests is low . Skin prick tests should always be done first . If there is a history of a previous serious reaction, the skin tests-if done-should be diluted to start with . Those doing penicillin skin tests should be prepared to treat a systemic reaction. Folia Microbiol (Praha), 1999, 44(6), 625 - 8 Escherichia coli strain with a deletion of the chromosomal ampC gene marked with TcR, suitable for production of penicillin G acylase; Vizvaryova M et al.; Escherichia coli strain which contains a marker of tetracycline resistance gene (TcR) placed by P1 transduction beside the chromosomal deletion of ampC gene (delta ampC) coding for beta-lactamase was constructed . Such introduction of TcR marker permits a fast and simple selection for the transfer of delta ampC by P1 transduction into industrial E . coli strains . This approach was used for constructing an E . coli strain suitable for penicillin acylase production. Clin Infect Dis, 2000 Dec, 31(6), 1512 - 4 Incidence of imipenem hypersensitivity reactions in febrile neutropenic bone marrow transplant patients with a history of penicillin allergy; McConnell SA et al.; The purpose of this retrospective study was to assess cross-hypersensitivity between imipenem/cilastatin and penicillin in patients with reported penicillin allergies . Medical records of febrile neutropenic, penicillin-allergic bone marrow transplant recipients who received imipenem/cilastatin treatment were retrospectively reviewed . The findings of this study indicate the incidence of cross-reactivity between imipenem/cilastatin and penicillin among patients with a history of penicillin allergy may be lower than previously reported. FEBS Lett, 2000 Nov 24, 485(2-3), 142 - 6 Studies of isopenicillin N synthase enzymatic properties using a continuous spectrophotometric assay; Dubus A et al.; Isopenicillin N synthase (IPNS) from Aspergillus nidulans is a no-heme iron(II)-dependent oxygenase which catalyses, in a single reaction, the bicyclisation of delta-(L-alpha-aminoadipoyl)-L-cysteinyl-D-valine into isopenicillin N, the precursor of all other penicillins, cephalosporins and cephamycins . The IPNS reaction can be followed directly and continuously by a new assay which monitors the absorbance increase at 235 nm characteristic of penicillin nucleus formation . Using this assay, the effects of influential factors affecting the in vitro IPNS enzymatic reaction were investigated . Even under optimal conditions, enzyme inactivation occurred during catalysis . Iron(II) depletion and product inhibition were not the cause of this phenomenon, the addition of antioxidants or reducing agents failed to slow down inactivation or reactivate the enzyme . Therefore, this phenomenon appears to be irreversible and is attributed to oxidative damage caused to the enzyme by reactive oxygen species generated in solution during catalysis . Nevertheless, the steady-state kinetic parameters for the IPNS reaction were determined. Curr Treat Options Neurol, 2000 Jul, 2(4), 375 - 387 Chronic Meningitis; Coyle PK; Treatment of chronic meningitis depends on the underlying cause . Once a specific cause has been established, appropriate targeted therapy is initiated . When the cause is unknown, a decision must be made whether to employ empiric therapy while the diagnostic evaluation is ongoing . This decision is based on three factors: 1) the clinical status of the patient; 2) the most likely cause based on demographic, historical, examination, and initial laboratory data; and 3) the risk-to-benefit ratio of the proposed treatment regimen . Initial efforts focus on deciding whether the meningitis is infectious or noninfectious . When a decision is made to start empiric therapy, antituberculous treatment is generally given for several weeks to judge therapeutic response . Patients who do not respond to antituberculous therapy are next treated with broad-spectrum antibiotics for unusual bacterial pathogens such as Actinomyces (penicillin), the Lyme disease spirochete Borrelia burgdorferi (ceftriaxone), Brucella (doxycycline plus rifampin), or Francisella tularensis (streptomycin plus gentamicin) . Antifungal therapy is generally reserved for patients who are at particular risk for mycotic infection (owing to underlying immunosuppression or an extraneural infection site) . Finally, empiric glucocorticoids are generally reserved for patients with consistently negative culture results and a probable noninfectious cause. Curr Treat Options Neurol, 1999 May, 1(2), 139 - 146 Neuroborreliosis (Nervous System Lyme Disease); Halperin JJ; Treatment of nervous system Lyme disease depends on the severity and site of involvement . Although some data indicate that uncomplicated Lyme meningitis can be treated effectively with oral doxycycline, central nervous system infection (meningitis, radiculitis, encephalomyelitis, and cranial neuritis) is usually treated with parenteral antibiotics for 14 to 30 days, depending on disease severity, as is severe and progressive peripheral nervous system involvement . Ceftriaxone, 2 g/d, is the most commonly used regimen; cefotaxime, 2 g every 8 hours, appears to be equally effective . Penicillin in meningeal doses is also effective, perhaps slightly less so than the third-generation cephalosporins, but it is less convenient to administer . For patients with prohibitive drug allergies, treatment with oral doxycycline in doses of 300 to 400 mg/d may be effective . In patients with facial palsy or with indolent peripheral neuropathies, a trial of oral medication (doxycycline, 100 mg two or three times a day, or amoxicillin, 500 to 1000 mg three times a day for 21 to 30 days) is reasonable . Patients for whom this fails are treated with parenteral medications. Curr Treat Options Cardiovasc Med, 1999 Oct, 1(3), 253 - 258 Rheumatic Fever; Visvanathan K et al.; There have been numerous reports stating that treatment of acute rheumatic fever with either aspirin or corticosteroids does not alter the long-term outcome of rheumatic heart disease . Yet, it should be emphasized that most of these studies were carried out with the first generic corticosteroids before the advent of the more active and more potent corticosteroid agents . In spite of this caveat, there is no question that all the clinical and laboratory parameters of inflammation (erythrocyte sedimentation rate, C-reactive protein) return to normal much more rapidly with corticosteroids than with aspirin alone . It is therefore our belief that steroids should be used when clinical and laboratory evidence of carditis exists, and aspirin should be reserved for cases of acute rheumatic arthritis with no evidence of carditis . The incidence of long-term valvular disease in active carditis may be decreased with steroid therapy . For example, the number of valve replacements differs markedly in centers that do use steroids and in those that do not . In Capetown, South Africa, where steroids are routinely used for carditis, valve replacement is quite rare . In contrast, in Johannesburg, where steroids are rarely used, the rate of valve replacement is quite high . The racial backgrounds of both groups of patients are similar, thus eliminating the question of racial differences . Concerning secondary prophylaxis, there is also controversy concerning the best second-line therapy . It is now well known that monthly intramuscular injections of benzathine penicillin are really effective for only 20 days . Thus, there is a window in which penicillin coverage is not adequate . To circumvent this problem, some investigators give benzathine penicillin every 3 weeks . These injections are quite painful, however, and it has been our "rule" that compliance with this treatment is inversely proportional to the ratio of the size of the child to the mother . In our own experience over 30 years with the follow-up of more than 300 patients with acute rheumatic fever, careful discussion of the consequences of missing oral doses has been adequate to insure proper compliance . An analysis of our patients on oral penicillin prophylaxis compared with other groups using benzathine penicillin revealed that there was very little difference in recurrence rate. Curr Infect Dis Rep, 1999 Apr, 1(1), 39 - 46 Deep Neck Infections; Nicklaus PJ; Deep neck infections are less common in the antibiotic era, but when they do occur they remain serious infections . Historically, these infections carried significant morbidity and mortality due to the proximity of the airway, mediastinum, and other vital structures . Deep neck infections were once routinely treated with penicillin, and if infection progressed to abscess, surgical drainage was performed . In recent years the standard medical, surgical, and diagnostic approaches to deep neck infection have changed, and they continue to evolve . Physicians must be aware of these changes to optimally manage patients with deep neck infections. Proc Natl Sci Counc Repub China B, 2000 Oct, 24(4), 156 - 60 Cultivation of recombinant Escherichia coli to achieve high cell density with a high level of penicillin G acylase activity; Liu YC et al.; A mutant strain of E . coli EP1 harbouring pGL-5 was employed to develop a process for producing penicillin G acylase (PGA) . In comparison with different carbon sources in the medium, it was found that the specific levels of PGA activity obtained in the glucose medium were the lowest . which was likely due to catabolic repression . Phenylacetic acid (PAA) was previously reported to be an regulatory inducer for PGA production, whereas in this study, the addition of PAA repressed both cell growth and enzyme expression . In a fed-batch culture, the increase of specific PGA activity followed the pattern of the cell concentration during the early to middle cell growth phase . With application of pure oxygen aeration and an appropriate medium design, the cell concentration reached 162 (g wet weight/l), which was 2.4 times higher compared to that of the original operation, and a specific PGA activity of 37 (IU/g wet weight) was achieved after 12 h of cultivation. J Biol Chem, 2001 Mar 2, 276(9), 6093 - 7 Epub 2000 Nov 14. The lysis protein E of phi X174 is a specific inhibitor of the MraY-catalyzed step in peptidoglycan synthesis; Bernhardt TG et al.; Coliphage phi X174 encodes a single lysis protein, E, a 91-amino acid membrane protein . Dominant mutations have been isolated in the host gene mraY that confer E resistance . mraY encodes translocase I, which catalyzes the formation of the first lipid intermediate in bacterial cell wall synthesis, suggesting a model in which E inhibits MraY and promotes cell lysis in a manner analogous to cell wall synthesis inhibitors like penicillin . To test this model biochemically, we monitored the effect of E on cell wall synthesis in vivo and in vitro . We find that expression of Emyc, encoding an epitope-tagged E protein, from a multicopy plasmid inhibits the incorporation of {(3)H}diaminopimelic acid into cell wall and leads to a profile of labeled precursors consistent with MraY inhibition . Moreover, we find that membranes isolated after Emyc expression are drastically reduced in MraY activity, whereas the activity of Rfe, an enzyme in the same superfamily, was unaffected . We therefore conclude that E is indeed a cell wall synthesis inhibitor and that this inhibition results from a specific block at the MraY-catalyzed step in the pathway. Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2000 Nov, 90(5), 636 - 40 Effect of systemic penicillin on pain in untreated irreversible pulpitis; Nagle D et al.; OBJECTIVE: The purpose of this prospective, randomized, double-blind study was to determine the effect of penicillin on pain in untreated teeth diagnosed with irreversible pulpitis.Study Design: Forty emergency patients participated, and each had a clinical diagnosis of an irreversible pulpitis . Patients randomly received a 7-day oral dose (28 capsules, 500 mg each, to be taken every 6 hours) of either penicillin or a placebo control in a double-blind manner . No endodontic treatment was performed . Each patient also received ibuprofen; acetaminophen with codeine (30 mg); and a 7-day diary to record pain, percussion pain, and number and type of pain medication taken . RESULTS: The administration of penicillin did not significantly (P >.05) reduce pain, percussion pain, or the number of analgesic medications taken by patients with untreated irreversible pulpitis . The majority of patients with untreated irreversible pulpitis had significant pain and required analgesics to manage this pain . CONCLUSION: Penicillin should not be prescribed for untreated irreversible pulpitis because penicillin is ineffective for pain relief. Mol Microbiol, 2000 Nov, 38(3), 658 - 65 G-protein signalling mediates differential production of toxic secondary metabolites; Tag A et al.; Filamentous fungi elaborate a complex array of secondary metabolites, including antibiotics and mycotoxins . As many of these compounds pose significant economic and health concerns, elucidation of the underlying cellular mechanisms that control their production is essential . Previous work revealed that synthesis of the carcinogenic mycotoxins sterigmatocystin (ST) and aflatoxin (AF) in Aspergillus species is negatively controlled by FadA, the alpha-subunit of a heterotrimeric G-protein . In sharp contrast, we show here that the dominant activating fadA allele, fadAG42R, stimulates transcription of a gene from the A . nidulans penicillin (PN) gene cluster and elevates penicillin production . Thus, FadA has opposite roles in regulating the biosynthesis of a potent antibiotic (PN) and a lethal mycotoxin (ST) in A . nidulans . Furthermore, expression of fadAG42R in Fusarium sporotrichioides increases trichothecene (TR) mycotoxin production and alters TR gene expression . Our findings reveal that a G-protein defines an important control point for differential expression of fungal secondary metabolites within and across fungal genera . These data provide critical evidence suggesting that targeting G-protein signal transduction pathways as a means of controlling or preventing the production of a single mycotoxin could have serious undesirable consequences with regard to the production of other secondary metabolites. Prescrire Int, 2000 Aug, 9(48), 106 - 9 Pneumococcal vaccination for elderly subjects: license extension . Still no proof of clinical efficacy; Homeostatic role of the active transport in elimination of {3H}benzylpenicillin out of the cerebrospinal fluid system; Department of Pharmacology, University of Zagreb School of Medicine, CroatiaCerebral acidic metabolites and penicillin are organic anions which can be carried by active transport into capillaries of the central nervous system (CNS) . However, it is generally believed that these metabolites are mainly delivered from CNS to cerebrospinal fluid (CSF) and eliminated by CSF circulation over cortex and its absorption into dural venous sinuses . To test this hypothesis we studied fate of penicillin ({3H}benzylpenicillin) in the CSF under control conditions and when its active transport was blocked by probenecid . After application of penicillin into cisterna magna of control dogs, it is distributed only in traces to lumbar, ventricular and cortical CSF . However, when active transport of penicillin across capillary wall is blocked by probenecid, its disappearance from cisterna is slowed down and its distribution is greatly enhanced so that at 300 min penicillin concentrations in cisternal, lumbar and cortical CSF approach or equal each other . Disappearance of penicillin from cisternal CSF shows a single exponential course (half-time 30 min) in control, while in probenecid pretreated dogs this is a slow multiexponential process . The results indicate that the active transport across capillary wall in CNS, but not generally postulated unidirectional CSF circulation over cortex and its absorption into dural venous sinuses, is instrumental in elimination of cerebral acidic metabolites and in such a way homeostasis in brain and cerebrospinal fluid is maintained. Prog Urol, 2000 Sep, 10(4), 587 - 9 {Renal actinomycosis with fistulized lumbar abscess}; Mallick S et al.; The authors report a case of renal actinomycosis in an adolescent presenting with two fistulized lumbar abscesses . This rare disease, which generally has a good prognosis, is difficult to diagnose, both clinically and radiologically . The positive diagnosis is based on histological examination, more frequently of the nephrectomy operative specimen, than ultrasound-guided fine needle aspiration biopsy of an atypical renal tumour . Conservative treatment with high-dose penicillin gives excellent results. Allergy Asthma Proc, 2000 Sep-Oct, 21(5), 297 - 9 The value of skin testing for penicillin allergy in an inpatient population: analysis of the subsequent patient management; Warrington RJ et al.; It was decided to assess the value of skin testing in a group of inpatients with a remote history of penicillin allergy, in terms of whether or not beta-lactams were subsequently given, if any adverse reactions occurred as a result of this therapy, and if labeling of the patient record was changed subsequent to skin testing and/or challenge . All patients seen in consultation with a history of penicillin allergy were assessed . When done, skin tests were performed with the major and minor determinants of penicillin and semisynthetic penicillins . Charts were reviewed after discharge in terms of the antibiotics given during admission, adverse events, and the medical record and hospital database labeling for drug allergy at discharge . Skin testing was carried out in 79% of 67 patients assessed and in all, the tests were negative . Beta-lactam therapy was recommended in 51/53 patients but was given in only 57% of these cases . At discharge, 49% of patients' records still carried the penicillin allergy label, despite negative skin testing and/or successful completion of a course of beta-lactam therapy . So, in approximately half of the patients reviewed, beta-lactams were not given despite negative skin tests and a recommendation to do so, if indicated, and 49% of patients were still inappropriately labeled as being penicillin-allergic on discharge. Bratisl Lek Listy, 2000, 101(7), 402 - 8 {Tularemia--an old and a new problem in the South Moravia Region}; Cerny Z; BACKGROUND: The first epidemic of ulceroglandular forms of tularemia acquired in coincidence with the manipulation with tularaemic hares took place in 1936 in the surroundings of Breclav and Valtice . The largest epidemic took place in the 1960's when hundreds of agricultural workers in the initial phases of the production of sugar within sugar refineries were afflicted by pulmonary forms of this disease . In the subsequent period which was interrupted only by smaller local epidemics, the number of new cases were gradually decreasing to the minimum at the beginning of 1990's . However, since 1994, the number of cases has began to increase again, namely those afflicted by ulceroglandular and oroglandular forms . SUBJECTIVES: In consequence of the long absence of this disease in clinical practice, the diagnostic awareness has decreased, and therefore the author has decided to indicate and review the current basic data on epidemiology and clinical manifestations of tularaemia . GROUP OF PATIENTS AND METHODS: The author has analysed the documentation of 577 of adults afflicted by tularemia and medically treated at the clinic of the Faculty Hospital in Brno in the period form 1959 to 1999 . The study reviews the onset of the disease and the pathway of transmission of infection and its clinical manifestation . MAIN RESULTS: Following the long-termed sporadic occurrence of tularaemia after major epidemics of pulmonary forms of this disease in 1960's, interrupted only by smaller local epidemics, the incidence has began to increase again in 1994 . The number of pulmonary forms has decreased whereas the occurrence of ulcerulceroglandular and oroglandular forms has increased . Hares have become the source of infection again . CONCLUSIONS: The fact that tularemia has repeatedly become a threat in Southern Moravia should be taken into account in the assessment of diagnosis in cases with unclear lymphadenitis and febrile states which defy the penicillin treatment especially in winter . (Tab . 3, Fig . 3, Ref . 23.) Rev Inst Med Trop Sao Paulo, 2000 Sep-Oct, 42(5), 287 - 9 Massive ocular hemorrhage resulting in blindness in a patient with the sickle cell trait who developed leptospirosis . Case report; Costa E et al.; This case report describes the findings of a 18 year-old black male from Bahia, a Northeastern state in Brazil, with the sickle cell trait, who developed bilateral hyphema and vitreous hemorrhage with blindness in the course of leptospirosis . The patient started to complain of blurred vision four days after the start of fever and muscular pain and approximately twelve hours after the introduction of penicillin . The severity of the leptospirosis in conjunction with sickle cell trait was considered to be the most likely explanation for this ocular complication. Ophthalmology, 2000 Nov, 107(11), 2015 - 23 Posterior segment manifestations of active ocular syphilis, their response to a neurosyphilis regimen of penicillin therapy, and the influence of human immunodeficiency virus status on response; Browning DJ; OBJECTIVE: To determine the relative frequencies of signs in posterior segment ocular syphilis, the response to a neurosyphilis regimen of penicillin, and differences in findings between human immunodeficiency virus (HIV)-coinfected and -noncoinfected patients in a community setting . DESIGN: Retrospective, noncomparative, consecutive case series . PARTICIPANTS: Fourteen consecutive patients with posterior segment ocular syphilis over a 14-year period within or during the acquired immune deficiency syndrome era . INTERVENTION: Neurosyphilis intravenous penicillin regimen . MAIN OUTCOME MEASURES: Initial and final visual acuity; treponemal and nontreponemal serologic analyses; cerebrospinal fluid cell count, protein, and Venereal Disease Research Laboratory analyses; posterior segment signs; and relapses and recurrences . RESULTS: Blacks and males were predominantly affected . Five (36%) of patients were HIV coinfected, and ocular syphilis led to the HIV infection diagnosis in three . Four (29%) patients had received previous antibiotic therapy for primary or secondary syphilis, raising the suspicion of relapse . Two patients had negative nontreponemal serologic results . All patients responded rapidly to neurosyphilis therapy . One patient subsequently relapsed after neurosyphilis therapy, and a second was reinfected with recurrence of ocular involvement . One previously undescribed retinal manifestation was discovered: a sectorial retinochoroiditis with delayed retinal circulation in the involved area . CONCLUSIONS: Ocular syphilis is a form of neurosyphilis and requires neurosyphilis therapy regardless of when it develops after primary infection . Conventional syphilis staging is of little use in understanding ocular syphilis . A high suspicion for this diagnosis is appropriate, especially in poorer black males with posterior segment inflammatory disease . Human immunodeficiency virus coinfection with ocular syphilis is common, but does not affect response to a neurosyphilis regimen of penicillin in the short term . Awareness of the multiple presentations of posterior segment ocular syphilis will aid ophthalmologists in averting misdiagnosis or delayed diagnosis. Pediatr Dent, 2000 Sep-Oct, 22(5), 401 - 4 Adverse reaction to amoxicillin: a case report; da Fonseca MA; Penicillin is the drug that most often leads to allergic reactions and anaphylaxis . The incidence of adverse events triggered by penicillins is believed to be between 1% and 10% . Up to one-tenth of these episodes are life-threatening, with the most serious reactions occurring in patients with no history of allergy . The case of a 5 year, 3 month-old female who had a severe allergic reaction to amoxicillin prior to a dental appointment is described . The literature on penicillin hypersensitivity is reviewed and recommendations for management of an allergic reaction in the pediatric dental office are discussed. Ugeskr Laeger, 2000 Oct 2, 162(40), 5351 - 3 {Penicillin treatment of acute maxillary sinusitis in adults . A randomized, double-blind, placebo-controlled trial from general practice}; Hansen JG et al.; INTRODUCTION: The aim of the study was to compare the effectiveness of penicillin V with placebo in the treatment of adult patients with acute maxillary sinusitis (in general practice) . MATERIAL AND METHODS: The study was designed as a randomised, double-blind, placebo-controlled trial in 26 Danish general practices . The participants were 133 adult patients with acute maxillary sinusitis clinically diagnosed on maxillary pain and raised values of either C-reactive protein (CRP) or the erythrocyte sedimentation (ESR) rate . Main outcome measures were pain score and illness score, and CRP and ESR values after initiation of treatment . RESULTS: Penicillin V led to a better recovery than placebo . The difference in pain reduction was statistically significant three days after initiation of treatment, whereas no significant difference was found in the reduction in the sense of illness . At the end of the study, significantly more patients in the penicillin group were completely free of pain than in the placebo group . This difference was found only in patients with an initial pain score of more than three . The cure rate was 71% in the penicillin group and 37% in the placebo group . Significantly more patients treated with penicillin achieved normal CRP values than those receiving placebo, respectively 88% and 75% . CONCLUSION: Penicillin V is more effective than placebo in the treatment of acute maxillary sinusitis in adults in general practice, but only in patients with pronounced pain. Chest, 2000 Oct, 118(4), 1106 - 8 A pilot study of penicillin skin testing in patients with a history of penicillin allergy admitted to a medical ICU; Arroliga ME et al.; BACKGROUND: Penicillin skin testing is an accurate method to determine whether a person with a history of penicillin allergy is at risk of having an immediate reaction to penicillin . A patient with a negative reaction to a skin test may be able to use a penicillin compound safely, which could reduce the use of broad-spectrum antibiotics in this patient population . METHODS: We prospectively studied all patients with histories of penicillin allergy who were admitted to a medical ICU during a 3-month period and who received antibiotics . Skin testing was performed with benzylpenicilloyl polylysine and penicillin G . We determined the incidence of true allergy, the percentage of patients in whom antibiotic coverage was modified, and the safety of the test . RESULTS: Two hundred fifty-seven patients were admitted to the medical ICU of The Cleveland Clinic Foundation during the study period . Twenty-four patients (9%), labeled as penicillin allergic and receiving antibiotics, were enrolled . Three patients (13%, 3 of 21) gave histories of type I reaction to penicillin and were not skin tested . Twenty patients (95%, 20 of 21) had negative skin test reactions to penicillin and positive skin test reactions to histamine control . One patient (4%, 1 of 21) with negative skin test reactions to both penicillin and histamine control had a test dose challenge with piperacillin that was well tolerated . There were no adverse events . Antibiotic coverage was changed in 10 patients (48%) as a result of skin testing . CONCLUSION: Most patients with histories of allergy to penicillin have negative reactions to skin tests and may receive penicillin safely . Penicillin skin testing can be utilized as a safe and effective strategy to reduce the use of broad-spectrum antibiotics. Int J Pediatr Otorhinolaryngol, 2000 Oct 16, 55(3), 197 - 201 Alteration of clinical picture and treatment of pediatric acute otitis media over the past two decades; Joki-Erkkila VP et al.; The clinical picture of acute otitis media (AOM) has changed greatly over the last few decades: serious complications have almost disappeared but more and more children suffer from recurrent middle ear infections and prolonged silent effusion . In this retrospective study we registered all AOM attacks among children under 10 in two rural municipal areas of Finland during 12-month-periods in 1978-79 and 1994-95 . In addition to epidemiological data, the clinical picture and given treatments were recorded . Between study periods the number of children with recurrent AOM attacks increased heavily . The percentage of spontaneous otorrhea decreased from 6.0 to 3.3% (P=0.01) and the proportion of afebrile patients increased from 64.0 to 73.4% (P=0.002) . In 1978-79 there were significantly more cases of diagnostic symptoms lasting over 24 h . Prescribing penicillin-V as a primary treatment decreased from 80.2 to 10.5% in favor of broad-spectrum antibiotics . The primary treatment with penicillin-V was associated with a decreased risk of recurrences . Acute tympanocentesis was performed less often and the incidence of surgical treatments (adenoidectomy and/or tympanostomy tube insertion) doubled from 6.2 to 12.4% of the acute cases . Although the clinical picture of AOM has become milder, children are treated with wider spectrum antibiotics . Nevertheless, a large number of children suffer from sequelae and there is a greater need for surgical treatments than 20 years ago. Ann Allergy Asthma Immunol, 2000 Sep, 85(3), 195 - 9 Penicillin allergy: prevalence of vague history in skin test-positive patients; Solensky R et al.; OBJECTIVE: Penicillin (PCN) skin testing is a reliable tool for predicting which patients can safely receive the antibiotic . Depending on the study, 7% to 76% of patients who have a history of PCN allergy have positive PCN skin tests . Many physicians approach patients who have a vague history of PCN allergy less cautiously than they approach those who have a convincing history suggestive of an IgE-mediated reaction . We reviewed the published literature to determine how many patients who had a history of PCN allergy and who were skin test-positive had a vague history of allergy . DATA SOURCES: By cross-referencing the keywords "penicillin" and "skin test," an Ovid MEDLINE search for English language studies published from 1966 to 1998 was performed . STUDY SELECTION: Studies in which history positive/skin test-positive patients were identified, and which contained documentation of the type of previous reaction in these patients, were included in the analysis . The MEDLINE search revealed 295 English language articles, of which 27 fulfilled the inclusion criteria . Three additional studies published prior to 1966 (and therefore not available through MEDLINE) also were found, bringing the total to 30 . A "convincing" history was defined to be one likely to be IgE-mediated (such as anaphylaxis, urticaria, angioedema or pruritic rash) . A "vague" history was one unlikely to be IgE-mediated (such as maculopapular rash, GI symptoms, or an unknown reaction) . RESULTS: Overall, 347/1063, or 33%, of history positive/skin test-positive patients had a vague PCN allergy history, with a range of 0% to 70% among the 30 studies . CONCLUSION: A large proportion of patients who have PCN-specific IgE antibodies, as determined by skin testing, have vague PCN allergy histories . These results therefore, indicate that, like patients with convincing histories, patients with vague allergic histories should undergo PCN skin testing prior to PCN administration. Prim, Care Update Ob Gyns . 2000 Sep 1, 7(5), 186 - 191 Syphilis in women; Edwards RK; Infection with the spirochete Treponema pallidum causes syphilis . Transmission of syphilis occurs through sexual contact with persons who have infectious mucocutaneous lesions . Before the advent of penicillin, this infection was responsible for a large portion of the debilitated patients residing in mental institutions . Later during this century, the disease became somewhat uncommon, only to see a resurgence during the early part of this decade . Syphilis progresses through stages defined as primary, secondary, latent, and tertiary . Diagnosis of this infection is by clinical examination, dark-field microscopy, and serology . Penicillin is the treatment of choice for all stages of syphilis . Doxycycline and tetracycline are acceptable alternatives in some penicillin-allergic patients . If patients are pregnant or have central nervous system involvement, alternative regimens should not be used because of lack of efficacy and/or fetal toxicity . Therefore, these patients must be desensitized and treated with penicillin . No effective vaccine for this infection is currently available. Appl Environ Microbiol, 2000 Oct, 66(10), 4536 - 8 Sulfur regulation of the sulfate transporter genes sutA and sutB in Penicillium chrysogenum; van de Kamp M et al.; Penicillium chrysogenum uses sulfate as a source of sulfur for the biosynthesis of penicillin . Sulfate uptake and the mRNA levels of the sulfate transporter-encoding sutB and sutA genes are all reduced by high sulfate concentrations and are elevated by sulfate starvation . In a high-penicillin-yielding strain, sutB is effectively transcribed even in the presence of excess sulfate . This deregulation may facilitate the efficient incorporation of sulfur into cysteine and penicillin. Korean J Parasitol, 2000 Sep, 38(3), 173 - 5 Postmetacercarial changes in Echinostoma caproni maintained in a defined medium plus calf serum; Fried B et al.; The present study examined postmetacercarial changes in the excysted metacercariae of Echinostoma caproni maintained in the defined medium Mixture 199 plus 20% calf serum for 7 days at 41 degrees C . The gas phase was atmospheric air . Each culture was inoculated with 25 excysted metacerariae . Cultures were maintained upright in closed 15 ml plastic centrifuge tubes each containing 10 ml of medium plus 200 units of penicillin/ml and 200 micrograms of streptomycin/ml . By 4 days in culture, most metacercariae had voided their excretory concretions . Organisms were clumped or solitary at the bottom of the cultures . Many organisms showed flaring of the oral collar and extension of both the collar and tegumentary spines . By 4 days in culture, posterior protuberances or bumps were noted on many of the organisms and some organisms showed abnormal vesicular growths or blebs at their posterior ends . Some mortality was noted in culture by day 5, but most organisms were still alive when the cultures were terminated on day 7. J Neuropsychiatry Clin Neurosci, 2000 Spring, 12(2), 246 - 50 Relationship between MRI findings and prognosis for patients with general paresis; Kodama K et al.; MRI was performed in 7 patients with general paresis before or at a very early stage of treatment . A large dose of antibiotics, mainly penicillin, was given to all patients, and the effects of treatment, the patients' outcome, and MRI findings were investigated . Three of the 7 patients had MRI findings of atrophy of the medial temporal lobe including the hippocampus . In the medial temporal lobe atrophy group, a personality change or general dementia remained even after the treatment was completed, and outcome in social functioning was poor . Medial temporal lobe atrophy may be a poor prognostic sign in general paresis. Clin Exp Allergy, 2000 Oct, 30(10), 1450 - 9 Characterization of primary recall in vitro lymphocyte responses to bacampicillin in allergic subjects; Cederbrant K et al.; BACKGROUND: Antigen-specific cell lines or clones are often used as models of drug-specific allergy . However, cloning procedures are time consuming, and the repeated antigen stimulation cycles as well as the addition of various growth enhancers may affect the in vivo relevance of these systems . OBJECTIVE: Using bacampicillin-allergic subjects, we wanted to investigate the applicability of primary recall in vitro lymphocyte responses to characterize type I and type IV allergy . The sensitivity and specificity of LTT (Lymphocyte transformation test), when used as an in vitro diagnostic tool, were also assessed . METHODS: A total of 39 patients with symptoms of type I (rhinitis) or type IV (allergic contact dermatitis, ACD) allergy following occupational exposure to bacampicillin, were included . Ten individuals without penicillin allergy or occupational exposure to bacampicillin served as controls . All subjects were LTT tested . Four patients with rhinitis and two patients with ACD were available for studying the immunophenotype and the TCR-Vbeta repertoire of bacampicillin induced lymphoblasts as well as the cytokine profiles and expression of the activation markers CD23 and CD134 in primary PBMC cultures . RESULTS: LTT was positive in 87% and at least one of the skin tests was positive in 85% of the patients with allergic symptoms . 69% of the patients with type I allergies were patch test-positive . Results from LTT and skin test correlated in 87% of the cases . The combined sensitivity of LTT and skin tests was 92% . The specificity of LTT was 90% in healthy controls . Bacampicillin induced lymphoblasts were mainly CD4 + in both ACD and rhinitis patients . The TCR-Vbeta profiles of the predominant CD4 + lymphoblasts were heterogeneous with individual skewing towards Vbeta2, Vbeta3, Vbeta5.1 and/or Vbeta14 . An increased expression of IFNgamma was detected in bacampicillin treated PBMC cultures from the ACD but not from rhinitis patients . IL-5 was detected in bacampicillin exposed PBMC cultures from all patients but not from healthy controls . This Th2 environment could also be verified by CD23 and CD134 expression . CONCLUSION: LTT and skin tests are equally sensitive in identifying bacampicillin allergic subjects . When the two tests are combined, the sensitivity increases . The patch test is useful not only for detection of type IV but also for the identification of type I allergies . When using primary PBMC cultures, IFNgamma is the most suitable cytokine to discriminate between type I and type IV allergy . IL-5 can possibly be used as a general marker for bacampicillin induced allergy . Thus, primary cell cultures may be considered as an alternative to T-cell lines or clones for the study of drug induced allergy. Clin Neurophysiol, 2000 Sep, 111 Suppl 2, S27 - 38 Spike-and-wave discharges of absence seizures as a transformation of sleep spindles: the continuing development of a hypothesis; Kostopoulos GK; OBJECTIVES: This review aims to offer a critical account of recent scientific developments relevant to the hypothesis which Pierre Gloor proposed in the 1970s for the generation of spike and wave discharges (SWDs) of primary generalized absence seizures . RESULTS: According to this hypothesis SWDs develop in the same circuits, which normally generate sleep spindles, by an initially cortical transformation of one every two or more spindle waves to a 'spike' component of SWDs, while the next one or more spindle waves are eliminated and replaced by a slow negative wave . This hypothesis was based on experiments in feline generalized penicillin epilepsy showing the possibility of transition from spindles to SWDs, when cortical neurons become hyper-responsive to thalamocortical volleys, which normally induce spindles, and thus engage feedback cortical inhibition, rebound excitation, recurrent intracortical dissemination of excitation during the 'spike' and strong excitation of thalamus for further augmentation of a brain wide synchronous oscillation . In the 1980s, electrophysiological studies in vitro and in vivo revealed the basic features of spindle rhythm generation by neurons in nucleus reticularis thalami and thalamocortical-corticothalamic oscillatory reverberations . CONCLUSIONS: In the light of this knowledge, experimental studies in several genetic and pharmacological animal models of absence seizures, clinical observations and theoretical studies in computer models have considered, tested, modified and challenged this hypothesis . It may still be found useful in the era of dynamic digital EEG analysis of SWDs and its current sources. Clin Neurophysiol, 2000 Sep, 111 Suppl 2, S9 - S18 Physiological basis: how NREM sleep components can promote and REM sleep components can suppress seizure discharge propagation; Shouse MN et al.; OBJECTIVES: To describe how the neural generators of different sleep components can provoke seizure discharge propagation during NREM sleep and can suppress it during REM sleep . METHODS: Experimental manipulations of discrete physiological components were conducted in feline epilepsy models (n=64), mostly in the systemic penicillin epilepsy model of primary generalized epilepsy and the amygdala kindling model of the localization-related seizure disorder, temporal lobe epilepsy . Procedures included seizure induction as well as quantifying norepinephrine concentrations (microdialysis) and the sleep-waking state distribution of seizures before and after lesions, systemic and localized drug administration and/or photic stimulation . RESULTS: (1) Neural generators of synchronous EEG oscillations, including tonic background slow waves and phasic 'arousal' events (sleep EEG transients such as sleep spindles, k-complexes), can combine to promote electrographic seizure propagation during NREM and drowsiness; anti-gravity muscle tone permits seizure-related movement . (2) Neural generators of asynchronous neuronal discharge patterns can reduce electrographic seizures during alert waking and REM sleep; skeletal motor paralysis blocks seizure-related movement during REM . (3) Etiology of the seizure disorder can interact with sleep and arousal mechanisms to determine sleep-waking state distribution of interictal and ictal events . CONCLUSIONS: Differential effects of NREM versus REM sleep components on seizure discharge propagation are to some extent non-specific and in other ways specific to seizure etiology. J Mol Biol, 2000 Sep 29, 302(4), 917 - 26 A fast method to predict protein interaction sites from sequences; Gallet X et al.; A simple method for predicting residues involved in protein interaction sites is proposed . In the absence of any structural report, the procedure identifies linear stretches of sequences as "receptor-binding domains" (RBDs) by analysing hydrophobicity distribution . The sequences of two databases of non-homologous interaction sites eliciting various biological activities were tested; 59-80 % were detected as RBDs . A statistical analysis of amino acid frequencies was carried out in known interaction sites and in predicted RBDs . RBDs were predicted from the 80,000 sequences of the Swissprot database . In both cases, arginine is the most frequently occurring residue . The RBD procedure can also detect residues involved in specific interaction sites such as the DNA-binding (95 % detected) and Ca-binding domains (83 % detected) . We report two recent analyses; from the prediction of RBDs in sequences to the experimental demonstration of the functional activities . The examples concern a retroviral Gag protein and a penicillin-binding protein . We support that this method is a quick way to predict protein interaction sites from sequences and is helpful for guiding experiments such as site-specific mutageneses, two-hybrid systems or the synthesis of inhibitors . J Mol Biol, 2000 Sep 29, 302(4), 887 - 98 Structure of a slow processing precursor penicillin acylase from Escherichia coli reveals the linker peptide blocking the active-site cleft; Hewitt L et al.; Penicillin G acylase is a periplasmic protein, cytoplasmically expressed as a precursor polypeptide comprising a signal sequence, the A and B chains of the mature enzyme (209 and 557 residues respectively) joined by a spacer peptide of 54 amino acid residues . The wild-type AB heterodimer is produced by proteolytic removal of this spacer in the periplasm . The first step in processing is believed to be autocatalytic hydrolysis of the peptide bond between the C-terminal residue of the spacer and the active-site serine residue at the N terminus of the B chain . We have determined the crystal structure of a slowly processing precursor mutant (Thr263Gly) of penicillin G acylase from Escherichia coli, which reveals that the spacer peptide blocks the entrance to the active-site cleft consistent with an autocatalytic mechanism of maturation . In this mutant precursor there is, however, an unexpected cleavage at a site four residues from the active-site serine residue . Analyses of the stereochemistry of the 260-261 bond seen to be cleaved in this precursor structure and of the 263-264 peptide bond have suggested factors that may govern the autocatalytic mechanism . Pediatr Dermatol, 2000 Jul-Aug, 17(4), 302 - 3 Partial lipoatrophy in a child; Kuperman-Beade M et al.; A 2-year-old boy was referred to the pediatric dermatology clinic for evaluation of an idiopathic, hypopigmented, atrophic patch on the right gluteal area . After extensive questioning the patient's parents recalled a penicillin injection several months prior to the appearance of lipoatrophy . There were no associated abnormalities. Structure Fold Des, 2000 Sep 15, 8(9), 971 - 80 Crystal structure of a D-aminopeptidase from Ochrobactrum anthropi, a new member of the 'penicillin-recognizing enzyme' family; Bompard-Gilles C et al.; BACKGROUND: beta-Lactam compounds are the most widely used antibiotics . They inactivate bacterial DD-transpeptidases, also called penicillin-binding proteins (PBPs), involved in cell-wall biosynthesis . The most common bacterial resistance mechanism against beta-lactam compounds is the synthesis of beta-lactamases that hydrolyse beta-lactam rings . These enzymes are believed to have evolved from cell-wall DD-peptidases . Understanding the biochemical and mechanistic features of the beta-lactam targets is crucial because of the increasing number of resistant bacteria . DAP is a D-aminopeptidase produced by Ochrobactrum anthropi . It is inhibited by various beta-lactam compounds and shares approximately 25% sequence identity with the R61 DD-carboxypeptidase and the class C beta-lactamases . RESULTS: The crystal structure of DAP has been determined to 1.9 A resolution using the multiple isomorphous replacement (MIR) method . The enzyme folds into three domains, A, B and C . Domain A, which contains conserved catalytic residues, has the classical fold of serine beta-lactamases, whereas domains B and C are both antiparallel eight-stranded beta barrels . A loop of domain C protrudes into the substrate-binding site of the enzyme . CONCLUSIONS: Comparison of the biochemical properties and the structure of DAP with PBPs and serine beta-lactamases shows that although the catalytic site of the enzyme is very similar to that of beta-lactamases, its substrate and inhibitor specificity rests on residues of domain C . DAP is a new member of the family of penicillin-recognizing proteins (PRPs) and, at the present time, its enzymatic specificity is clearly unique. J Immunol Methods, 2000 Aug 28, 242(1-2), 43 - 51 In vitro reactivity of penicilloyl and penicillanyl albumin and polylysine conjugates with IgE-antibody; Zhao Z et al.; Penicilloylated (BPO) and penicillanylated (BPA) poly-L-lysine (PLL) and human serum albumin (HSA) were prepared and characterized by penamaldate assay and proton NMR spectroscopy . The conjugates were coupled to nitrocellulose (NC) discs and cyanogen bromide activated paper discs and their in vitro reactivities with serum IgE antibodies were examined . Results showed that on paper discs, 55.3 and 83% of the sera reacted with PLL conjugates of BPO and BPA, respectively, while 41.5 and 58.1% reacted with HSA conjugates . On NC discs, HSA conjugates gave better results, 75.6 and 70.7%, respectively for BPO and BPA, compared with 38.6 and 50%, respectively for the PLL conjugates . Overall, the BPA-PLL conjugate on paper discs proved to be the most reactive preparation . Addition of the BPO-PLL paper disc preparation detected more positive sera (85.1%) and we believe that the combined use of these two specificities offers the best test for the detection of penicillin-reactive IgE antibodies. Indian J Biochem Biophys, 2000 Feb, 37(1), 6 - 12 Penicillin acylase catalyzed synthesis of penicillin-G from substrates anchored in cyclodextrins; Prabhu KS et al.; Penicillin acylase (EC 3.5.1.11) catalyses the condensation of phenylacetic acid (PAA) and 6-aminopenicillanic acid (6-APA) to form benzylpenicillin (BP) . Both PAA and 6-APA were found to form host-guest complexes with beta-methylcyclodextrin (beta m-CD) and gamma-cyclodextrin (gamma-CD) respectively . The rate of the reaction catalyzed by the enzyme remained unaffected if one of the substrates used was in the cyclodextrin complexed form . However, in this case, the reaction lasted longer and yielded about 20 per cent more products compared to the condensation reaction involving only uncomplexed substrates . There was distinct increase in the rate of formation of the antibiotic, if both substrates used are in CD-complexed form. Forensic Sci Int, 2000 Sep 11, 113(1-3), 497 - 9 An historical review of criminal abortion; Williams DJ et al.; Criminal abortion is rarely seen in modern Australia . This historical review examine cases seen in Brisbane during the period 1939 and 1947 . Instrumentation or introduction of fluid into the uterus were the common causes of fatalities in this pre-penicillin era. Bull Exp Biol Med, 2000 May, 129(5), 420 - 2 Effect of akatinol (memantine) in central spinal pain syndrome; Zinkevich VA et al.; On the model of central spinal pain syndrome in rats induced by application of penicillin to the dorsal surface of the lumbar spinal cord, akatinol injected intraperitoneally at the peak of syndrome or applied locally simultaneously with