Mol Microbiol, 1995 Oct, 18(1), 123 - 33 Cation-selectivity of the L-glutamate transporters of Escherichia coli, Bacillus stearothermophilus and Bacillus caldotenax: dependence on the environment in which the proteins are expressed; Tolner B et al.; L-Glutamate transport by the H(+)-glutamate and Na(+)-glutamate symport proteins of Escherichia coli K-12 (GltPEc and GltSEc, respectively) and the Na(+)-H(+)-glutamate symport proteins of Bacillus stearothermophilus (GltTBs) and Bacillus caldotenax (GltTBc) was studied in membrane vesicles derived from cells in which the proteins were either homologously or heterologously expressed . Substrate and inhibitor specificity studies indicate that GltPEc, GltTBs and GltTBc fall into the same group of transporters, whereas GltSEc is distinctly different from the others . Also, the cation specificity of GltSEc is different; GltSEc transported L-glutamate with (at least) two Na+, whereas GltPEc, GltTBs and GltTBc catalysed an electrogenic symport of L-glutamate with > or = two H+, i.e . when the proteins were expressed in E . coli . Surprisingly studies in membrane vesicles of B . stearothermophilus and B . caldotenax indicated a Na(+)-H(+)-L-glutamate symport for both GltTBs and GltTBc . The Na+ dependency of the GltT transporters in the Bacillus strains increased with temperature . These observations suggest that the conformation of the transport proteins in the E . coli and the Bacillus membranes differs, which influences the coupling ion selectivity. Proteins, 1995 Oct, 23(2), 290 - 3 Crystallization of a membrane pore-forming protein with mosquitocidal activity from Bacillus thuringiensis subspecies kyushuensis; Li J et al.; CytB, a membrane pore-forming toxin from Bacillus thuringiensis subspecies kyushuensis, is specifically toxic to dipteran insect larvae but broadly cytolytic in vitro . It has been purified in the protoxin form from a recombinant Escherichia coli source and crystals have been obtained which diffract X-rays to at least 2.6 A resolution . The tendency for CytB to aggregate in solution was overcome by including 50 mM of urea or 8 mM of ethanolamine during crystallization . Mutants designed to add or subtract single cysteine residues for the purpose of heavy atom derivative preparation were similarly purified and crystallized . The crystals are hexagonal bipyramids . They belong to space group P6(1)22 (or P6(5)22) with lattice constants a = b = 67.34 A, c = 170.96 A, and contain one molecule of the CytB protoxin (MW 29235) per asymmetric unit and 27% solvent by volume. Med Parazitol (Mosk), 1995 Oct-Dec, (4), 49 - 53 {The outlook for improving the immunoglobulin preparations for the detection and identification of the causative agents of glanders and melioidosis}; Khrapova NP et al.; The paper summarizes the data concerning the production and study of monoclonal antibodies (MAb) to the diagnostically significant glanders and melioidosis bacillus antigens . It evaluates the efficiency of using MAb in the gel immunodiffusion and agglutination tests as a basis of new-generation preparations for fluorescent antibody assay, indirect hemagglutination test which are used while detecting and identifying pathogenic pseudomonads . The paper defines the quality indices for monoclonal luminescent immunoglobulins and provides evidence for the benefits of monoclonal diagnostic agents over polyclonal analogues. Med Parazitol (Mosk), 1995 Oct-Dec, (4), 42 - 5 {Brucellosis in Siberia and the Far East}; Kalinovskii AI et al.; Epidemiological analysis has indicated that rat and reindeer brucellosis foci are of definite value in Siberia and the Far East in the liquidation of brucellosis ones . Foci of cattle and reindeer have been first established, evidence has been provided for the epidemiological significance of fifth-biological variant B . ovis and B . suis, as well as the ecological confinement of peculiar B . rangiferi cultures to the brucellosis foci in the Arctic . To plan antibrucellosis efforts, it is necessary to take into account the incidence of human infection, as well as the insidious circulation of the bacillus in the stock farms. Med Parazitol (Mosk), 1995 Oct-Dec, (4), 3 - 9 {The problem of natural foci of plague: the search for ways for its resolution}; Lomaradskii IV et al.; The paper deals with one of the most important problems of plague natural foci, namely the places and types of its bacillus persistence outside the parasitic system, as well as in the interepizootic period . It also discusses different hypotheses forwarded on the basis of field observations and experimental findings . In particular, it gives the materials suggesting that the plague bacillus can persist in the low-virulent form under natural conditions, as well as due to its transition to a L-form . The paper provides strong evidence for the long-term persistence of the plague bacillus in soil and hole substrates and discloses the adaptive mechanisms ensuring such persistence . Particular emphasis is placed on the complex symbiosis of the plague bacillus with soil microorganisms, including protozoa, and on their persistence in cysts of the latters . Tasks and a trend in further studies required to finally solve the problem are stated in the paper. Med Parazitol (Mosk), 1995 Oct-Dec, (4), 27 - 30 {THe characteristics of the epidemic process of pseudotuberculosis infection in the southern and northwestern foci of Irkutsk Province}; Chesnokova MV et al.; A long-term study of pseudotuberculosis epidemiology has revealed two independent foci in the south and northwest of Irkutsk Province, which differ in the level and pattern of morbidity, seasonal features, patients' age distribution, severity of a clinical course . Each focus is characterized by the clonal structure of the pseudotuberculosis bacillus, which is persistent in terms of the plasmid profile . Strains with the two plasmids 47 mD (pYV) and 82 mD (pVM) circulate in the southern focus, while those with one plasmid 47 mD (pYV) in the northwestern focus . The intensity of an epidemic process is not associated with the presence of the plasmid 82 mD (pVM) . The presence of the latter statistically significantly determines the severity of the clinical course of pseudotuberculosis infection, which seems to be caused by the immunodepressive action of the plasmid pVM. Med Parazitol (Mosk), 1995 Oct-Dec, (4), 14 - 7 {An experimental study of the ability of the hybrid crosses of Citellophillus tesquorum fleas of 2 subspecies to transmit the causative agent of plague}; Nikitin AIa et al.; The ability to transmit of plague bacillus was studied in the first-, third- and fourth-generation hybrids bred from C.t.altaicus and C.t.sungaris . The fleas were inoculated with two virulent strains from the Tuva and Transbaikal natural foci of plague . The bacillus was demonstrated to infect the hybrids, form gizzard block, and be transmitted to white mice, causing the animals' death. Am J Infect Control, 1995 Oct, 23(5), 329 - 36 Development of a research-based protocol to rule out tuberculosis by means of continuous quality improvement techniques; Harmon JC et al.; Centers for Disease Control and Prevention guidelines mandate that patients be kept in respiratory isolation until three negative results of acid-fast bacillus smears, obtained on three separate days, have been documented . This mandate, coupled with the increased prevalence of Tuberculosis and a limited number of isolation rooms, indicated that an effective system needed to be developed to avoid unnecessary time spent in isolation . When it was found that patients spent a mean of 6.6 days in isolation before three negative results of acid-fast bacillus smears were obtained, a quality improvement project was initiated by the department of medicine . A multidisciplinary group with representation from epidemiology, nursing, medicine, the microbiology laboratory, and respiratory therapy reviewed current practice and developed a protocol to rule out tuberculosis . The protocol, which incorporates gastric lavage and sputum induction, facilitates the collection and processing of specimens within a target of 4 days from the time of placement in isolation . The protocol was reinforced by traditional staff education and large posters displayed on nursing units for easy reference . The mean time spent in isolation to rule out tuberculosis was significantly decreased with the protocol to 4.9 days (p < 0.001) . After implementation, 56.6% of patients were out of isolation within 4 days, compared with only 2.6% before the protocol's implementation . This represents both a significant cost saving and improved patient outcomes . Current efforts center on continuous maintenance and improvement of these gains. Int J Biol Macromol, 1995 Oct, 17(5), 259 - 67 Effects of manganese (II) on Bacillus licheniformis ATCC 9945A physiology and gamma-poly(glutamic acid) formation; Cromwick AM et al.; Bacillus licheniformis ATCC 9945A was cultivated in shake flasks using citrate (12 gl-1), glutamate (20 gl-1) and glycerol (80 gl-1) as carbon sources for cell growth and gamma-poly(glutamic acid) (gamma-PGA) production . The effect of the MnSO4 concentration in the medium over a range from 0.0 to 615 microns was studied . The number of viable cells increased for all concentrations of MnSO4 from approximately 10(5) to 10(9) colony-forming units (cfu) ml-1 by the early stationary phase (24 h) . However, after 50 h, the cell viability decreased rapidly for relatively lower MnSO4 concentrations (0.615 and 0 microns) . The utilization of carbon sources by B . licheniformis was greater for cultures containing 33.8 and 615 microns MnSO4 relative to cultures with no added MnSO4 . For example, cultures with 615 microns MnSO4 utilized 37, 54 and 93% and cultures with no added MnSO4 utilized 19, 10 and 17% of glutamate, glycerol and citrate, respectively . The gamma-PGA volumetric yield increased from approximately 5 to 17 gl-1 for corresponding increases in MnSO4 concentration from 0 to 33.8 microns and then decreased at higher MnSO4 concentrations . The stereochemical content of gamma-PGA was found to vary inversely with MnSO4 concentration, and ranged from 59 to 10% L-glutamate units for MnSO4 concentrations of 0 and 615 microns, respectively . For all of the MnSO4 concentrations investigated, the gamma-PGA molecular weights decreased rapidly as the gamma-PGA volumetric yield simultaneously increased for cultivation times from 24 to approximately 50 h . Mw and Mn values after approximately 50 h cultivation times, determined by gel permeation chromatography (GPC), were 1.3 to 1.6 and 0.5 to 0.8 million g mol-1, respectively . A complex gamma-PGA molecular weight distribution that appeared bimodal by GPC analysis due to the presence of a low-molecular-weight product fraction was observed in cultures containing 33.8 and 61.5 microns MnSO4 at extended cultivation times . A high-molecular-weight fraction and the unfractionated gamma-PGA sample from the 33.8 microns MnSO4 culture contained 13 +/- 4 and 30 +/- 1% L-repeat units, respectively . A relationship between the product molecular weight and its stereochemical composition was thus established. Int J Food Microbiol, 1995 Oct, 27(2-3), 175 - 83 Characteristics of some psychrotrophic Bacillus cereus isolates; Dufrenne J et al.; Twelve strains of Bacillus cereus isolated from different food products and foodborne disease outbreaks, and able to grow at temperatures < 7 degrees C, were characterised . Generation times at 7 degrees C varied from 9.4 h up to 75 h . Lag phase of the vegetative cells at 7 degrees C was strongly influenced by the previous temperature history of the cells . Preincubation at 37 degrees C increased the duration of the lag phase drastically . The heat resistance at 90 degrees C (D90 degrees C-values in min) for spores produced at 30 degrees C varied from 2.2 to 9.2 min for 11 strains . One strain, however, showed a D90 degrees C-value of > 100 min . Germination of spores in milk was delayed compared to those grown in brain heart infusion broth (BHI) . All strains showed production of the diarrheal type enterotoxin in BHI . Addition of 50 IU of nisin to skim milk resulted in a decrease of numbers for 9 of the 12 strains tested . At a nisin concentration of 250 IU, a decrease in bacterial numbers was observed for all strains tested. Exp Anim, 1995 Oct, 44(4), 333 - 6 Detection of cilia-associated respiratory bacillus in experimentally and naturally infected mice and rats by the polymerase chain reaction; Goto K et al.; Cilia-associated respiratory (CAR) bacillus was detected by means of the reverse transcription (RT)-polymerase chain reaction (PCR), and the results were compared with those of indirect immunofluorescence test (IFAT) for the detection of the organism . In the experimental infections, 15 mice were in contact with mice previously inoculated with CAR bacillus . Three mice each were tested at days 3, 5, 7, 12 and 20 postexposure . On day 3 postexposure, CAR bacillus was detected in oral swab samples from all 3 mice by RT-PCR, but was not detected in any sampling sites from the mice by IFAT . Total numbers of positive samples from nasal, oral and tracheal swabs obtained through the test were 6/15, 14/15 and 8/15, respectively, by RT-PCR, and 2/15, 6/15 and 3/15, respectively by IFAT . For the detection of CAR bacillus in samples from 52 rats, 34 serum antibody negative rats by enzyme-linked immunosorbent assay were also negative by RT-PCR and IFAT except for one sample from the oral cavity, and all serum antibody positive rats were positive for the organism by RT-PCR but it could not be detected in five of them by IFAT . By means of RT-PCR, no differences in the positive rates depending on sampling sites were observed except in one rat . The RT-PCR was found to be a specific, highly sensitive and reliable procedure for detecting CAR bacillus in mice and rats . The oral cavity was the most suitable site for the diagnosis of the early stage of this infection by RT-PCR. Diagn Cytopathol, 1995 Oct, 13(3), 225 - 8 Detached single cilia: another potential pseudomicrobe seen in bronchoalveolar lavage specimens; Stanley MW et al.; High magnification examination of Romanovsky-stained bronchoalveolar lavage (BAL) material may be needed, if one is to confidently exclude small organisms such as toxoplasmosis, histoplasmosis, and the negative images of mycobacteria . Pseudomicrobes have been described, and detailed criteria must be carefully applied, if incorrect diagnoses are to be avoided . We recently noted numerous free-lying, uniform, eosinophilic, straight or slightly curved bacilliform structures in such a specimen . These were seen on Diff-Quik-stained material, but were extremely pale on Papanicolaou-stained slides . Comparison with the cilia of contaminating bronchial cells showed these pseudomicrobes to have similar length, diameter, and tinctorial properties . Electron microscopy of the sedimented cell pellet showed isolated, detached single cilia (DSC) far removed from any bronchial cells . Twenty BAL's from 13 males and seven females (median age = 48.5) undergoing infection surveillance were then retrospectively reviewed . DSC were present in 16 specimens (80%), but were rare in three of these . Although present in cases with prominent bronchial cell contamination (n = 6 cases), DSC were also encountered when ciliated cells were rare (n = 13), or absent (n = 1) . Familiarity with these distinctive structures should make it unlikely that they will be mistaken for infectious organisms. Antonie Van Leeuwenhoek, 1995 Oct, 68(3), 203 - 8 Hexavalent-chromium reduction by a chromate-resistant Bacillus sp . strain; Campos J et al.; Bacillus strain QC1-2, isolated from a chromium-polluted zone, was selected by its high ability to both tolerate and reduce hexavalent chromium {Cr(VI)} to less-toxic trivalent chromium {Cr(III)} . Cell suspensions of strain QC1-2 rapidly reduced Cr(VI), in both aerobic and anaerobic conditions, to Cr(III) which remained in the supernatant . Cr(VI) reduction was dependent on the addition of glucose but sulfate, an inhibitor of chromate transport, had no effect . Studies with permeabilized cells and cell extracts showed that the Cr(VI) reductase of strain QC1-2 is a soluble NADH-dependent enzyme. J Electron Microsc (Tokyo), 1995 Oct, 44(5), 281 - 8 Ultrahigh-resolution scanning electron microscopy of MDCK cells infected with influenza viruses; Hayase Y et al.; Ultrahigh-resolution scanning electron microscopy of MDCK cells infected with influenza viruses was carried out by the uncoated uranyl acetate staining preparation method . Ridge-like protrusions were detected on virus particles and were considered to be an indication of aggregated glycoprotein spikes . Bundles of filamentous virus particles along with bacillary virus particles were encountered on MDCK cells infected with freshly isolated strains of type A virus . Filamentous virus particles that were twisted like ropes were observed on MDCK cells infected with strains of type B virus. Trends Microbiol, 1995 Oct, 3(10), 401 - 4 Prospects for new vaccines against tuberculosis; Orme IM; The spectacular failure of the existing bacille Calmette-Guerin (BCG) vaccine in recent tuberculosis-control trials has prompted a search for potential replacements . Imaginative approaches currently under development include the modulation of BCG by the production of auxotrophic mutants or cytokine-secreting recombinants, and the development of subunit vaccines based on the major proteins of Mycobacterium tuberculosis. Trends Microbiol, 1995 Oct, 3(10), 397 - 401 BCG vaccination against tuberculosis: past disappointments and future hopes; Roche PW et al.; Three billion doses of the bacille Calmette-Guerin (BCG) vaccine have been given over the past 60 years . BCG has a widely variable ability to protect humans against tuberculosis . This article examines some of the reasons for this variability, and discusses ways in which BCG vaccination might be improved. Int J Epidemiol, 1995 Oct, 24(5), 1042 - 9 Efficacy of BCG vaccination of the newborn: evaluation by a follow-up study of contacts in Bangui; Lanckriet C et al.; BACKGROUND . The efficacy of Bacillus of Calmette and Guerin (BCG) vaccination given at birth is still controversial . We therefore conducted a study in Bangui (Central African Republic) to estimate the protection afforded over the first seven years of life by BCG administered at birth . METHODS . One thousand children who had lived in contact with a recently diagnosed case of contagious tuberculosis were followed up for a period of 6 months in order to detect the occurrence of tuberculosis . Diagnosis of tuberculosis was made through a scoring system . Vaccine efficacy (VE) was calculated on the basis of the relative risk of contracting tuberculosis according to vaccination status . RESULTS . The efficacy of BCG was estimated to be 71% (95% confidence interval: 56-81%) . This result remained practically the same after changing the definition used for tuberculosis cases (VE = 75% for a threshold with a score of 15 instead of 6, VE = 74% when only confirmed cases were considered) . There was no difference between the two groups in the variables measuring intensity of contact with the source of contamination, but there was a difference in age distribution . Vaccine efficacy adjusted for this factor was the same as the crude VE . CONCLUSION . This study, based on a methodology that controls for most of the risks of bias inherent to field efficacy measurement, confirms the protective capacity of neonatal BCG against childhood tuberculosis . Therefore BCG vaccination at birth must remain a public health priority especially in countries with high incidence of the diseasePIP: The efficacy of Bacillus of Calmette and Guerin (BCG) vaccination given at birth is still controversial, therefore a study was conducted in Bangui, Central African Republic, to estimate the protection afforded over the first 7 years of life by BCG administered at birth . 1000 children who had lived in contact with a recently diagnosed case of contagious tuberculosis (TB) were followed up from May 1989 to February 1991 in order to detect the occurrence of TB . 896 of them were considered as vaccinated . Diagnosis of TB was made through a scoring system endorsed by the World Health Organization . Contact children with a score of or= 6 were considered to have TB . Four groups of children were formed in order to calculate the risk of TB in relation to vaccination status: children with TB who had been vaccinated, children with TB who had not been vaccinated, healthy children who had been vaccinated, and healthy children who had not been vaccinated . Vaccine efficacy (VE) was calculated on the basis of the relative risk of contracting TB according to vaccination status . Of the 1000 contact children, 91 had a score of or= 6 . The TB incidence rate was 7.3% in vaccinated children and 25% in nonvaccinated children, which corresponded to a 0.29 relative risk of contracting TB or efficacy of BCG of 71% (95%, confidence interval: 56-81%) . This result remained practically the same after changing the definition used for TB cases (VE = 75% for a threshold with a score of 15 instead of 6, VE = 74% when only confirmed cases were considered) . There was no difference between the 2 groups in the variables measuring intensity of contact with the source of contamination, but there was a difference in age distribution . BCG vaccination at birth must remain a public health priority, especially in countries with high incidence of the disease in view of the protective capacity of neonatal BCG against childhood TB . Skeletal Radiol, 1995 Oct, 24(7), 499 - 503 Musculoskeletal melioidosis: clinical and imaging features; Pui MH et al.; OBJECTIVE . Melioidosis is a tropical infection caused by a gram-negative bacillus, Pseudomonas pseudomallei . The disease manifests initially as localized suppurative lesions and can progress to acute disseminated septicemia with 65-90% mortality if inadequately treated . Musculoskeletal involvement is common . The purpose of this study was to describe the clinical features and imaging appearances of musculoskeletal melioidosis . DESIGN . We retrospectively analyzed the clinical profiles and images of 26 patients diagnosed over a 6-year period as suffering from melioidosis . PATIENTS . The study group comprised 11 patients with musculoskeletal melioidosis and 15 patients with nonmusculoskeletal melioidosis . RESULTS AND CONCLUSIONS . We found that musculoskeletal melioidosis mimicks other infections both clinically and radiologically . Clinical awareness is therefore crucial, as diagnosis can only be established by bacteriological and immunological studies . Prompt treatment with long-term combination antibiotics in high dosages and surgical drainage of abscesses improves survival. Biosci Biotechnol Biochem, 1995 Oct, 59(10), 1994 - 5 Nucleotide sequencing of phenylalanine dehydrogenase gene from Bacillus badius IAM 11059; Yamada A et al.; The nucleotide sequence of the pdh gene coding for PheDH from Bacillus badius IAM 11059 was analyzed . The gene consists of an ORF of 1140 nucleotides which specifies a protein of 380 codons . The primary structure of PheDH is similar to PheDH from B . sphaericus, PheDH from Themoactinomyces intermedius, and leucine dehydrogenase from B . stearothemophilus, etc. Biosci Biotechnol Biochem, 1995 Oct, 59(10), 1856 - 60 Isolation and characterization of a novel endo-beta-galactofuranosidase from Bacillus sp; Ramli N et al.; A soil bacterium capable of growing on a polysaccharide-containing beta(1-->6)galactofuranoside residues derived from the acidic polysaccharide of Fusarium sp . as a carbon source has been isolated . From various bacteriological characteristics, the organism was identified as a Bacillus sp . The bacterium produced beta-galactofuranosidase inductively in the culture media . The most effective inducer for the beta-galactofuranosidase production was a polysaccharide containing beta(1-->5) or beta(1-->6)-linked galactofuranoside residues, but gum arabic, gum guar, gum ghati, arabinogalactam, araban, and pectic acid did not induce the enzyme . The enzyme had three different molecular weight forms . The low molecular-weight form was purified by a combination of Toyopearl HW-55 and DEAE-Toyopearl 650S column chromatographies, and preparative polyacrylamide gel electrophoresis . The molecular weight of the enzyme was estimated to be 67,000 by SDS-polyacrylamide gel electrophoresis . The enzyme was most active at pH 6 and 37 degrees C, and was stable between pH 4 to 8 at 5 degrees C . The action of the enzyme was inhibited by the addition of Cd2+, Co2+, Hg2+, Zn2+, iodoacetic acid, and EDTA . The purified enzyme cleaved beta(1-->5) and beta(1-->6)-linked galactofuranosyl chains . Based upon the mode of liberation of galactofuranosyl residues from pyridylamino-beta(1-->6)-linked galactofuranoside oligomers, the enzyme can be classified as an endo-beta-galactofuranosidase that randomly hydrolyzes the linkage. Biosci Biotechnol Biochem, 1995 Oct, 59(10), 1817 - 24 Reddish Escherichia coli cells caused by overproduction of Bacillus stearothermophilus uroporphyrinogen III methylase: cloning, sequencing, and expression of the gene; Chou PL et al.; During shotgun cloning of an amylase gene, we found a transformant of Escherichia coli with a reddish color . The transformant produced highly water-soluble red pigments the molecular masses of which were less than 3000 . The plasmid harbored by the transformant contained a DNA fragment derived from a strain of Bacillus stearothermophilus . Truncation of the insert DNA showed that an 1.1-kbp Sau 3A-SalI fragment was responsible for the reddish colony . An open reading frame was found in the nucleotide sequence of the 1.1-kbp DNA fragment . The production of the red pigment was accompanied by a colorless 28-kDa protein . The sequence of the 28-kDa protein was highly homologous to bacterial uroporphyrinogen III methylases participating in corrinoid biosynthesis . The 28-kDa protein was found to be a thermostable uroporphyrinogen III methylase. Am J Ind Med, 1995 Oct, 28(4), 469 - 88 Cross-sectional follow-up of a flu-like respiratory illness among fiberglass manufacturing employees: endotoxin exposure associated with two distinct sequelae; Milton DK et al.; Over a period of 10 years, employees in a manufacturing plant experienced sporadic flu-like episodes after work in a basement containing a recirculated washwater mist . We report a cross-sectional study to define the flu-like illness and bioaerosol exposures . High concentrations of gram-negative bacteria (GNB) (> 10(7) cfu/ml) and endotoxin (range 34-46 micrograms/ml) were found in the water . Mist contained > 10(3) cfu/m3 of GNB, and endotoxin up to 13,900 to 27,800 ng/m3 . Few fungi and thermotolerant Bacillus species and no Actinomycetes, Legionella species, or amoeba were found in washwater . Airborne levels of fungi were of the same species and magnitudes as outdoor samples . Subjects volunteered (n = 28) because of a history of flu-like symptoms or were randomly selected (n = 102) from workers with and without current exposure to the basement . No acute cases were examined . Cases did not fulfill criteria for hypersensitivity pneumonitis (HP) and high levels of IgG antibodies to water-borne antigens were not observed . However, among 20 subjects indicating a history of severe flu-like episodes (severe basement flu, SBF), diffusion capacity (DLCO) was significantly lower (p = 0.015) than among other workers . The prevalence of SBF was independent of smoking . Cases occurred in clusters, and SBF was more common among workers with intermittent exposure to the basement (19 cases) than with daily exposure (1 case) . These findings suggest that SBF and associated chronically depressed DLCO resulted from toxic injury following high-level endotoxin exposure . Asthma was prevalent in the study population, particularly among employees with daily, rather than intermittent, exposure to endotoxin-containing mist (odds ratio 6.7, p = 0.02) . Thus, endotoxin exposure in this study was associated with two distinct sequelae depending on the temporal pattern of exposure. Arch Esp Urol, 1995 Oct, 48(8), 791 - 5 {Calmette-Guerin bacillus in the prophylaxis of superficial bladder tumors . Our experience and our doubts}; Moyano Calvo JL et al.; OBJECTIVE: This study was conducted to determine the efficacy of our BCG dose and schedule for prophylactic therapy in superficial bladder tumors . METHODS: Following complete TUR, 81 mg intravesical BCG were administered weekly for six weeks and for nightly another six times . The patients were evaluated every three months by cytology, cystoscopy and routine analysis . RESULTS: 21/25 patients completed the treatment . At 15.5 months median follow-up, 18/21 (85.7%) were disease-free, 2/21 (9.5%) had recurrence and 1/21 (4.7%) showed disease progression . CONCLUSIONS: BCG is the most effective prophylactic therapy for superficial bladder tumors currently available . The ideal dose, treatment schedule and its mechanism of action are as yet unknown. J Inorg Biochem, 1995 Oct, 60(1), 61 - 7 Synthesis, characterization and antibacterial activity of novel Fe(III), Co(II), and Zn(II) complexes with norfloxacin; Gao F et al.; Three novel complexes of norfloxacin (abbreviated as NFL), {M(NFL)2(H2O)2}Cl3.6H2O, (M = Fe, Co), and {Zn(NFL)2}Cl2.7H2O, have been prepared . The compounds were characterized by IR, UV-Vis, NMR spectra, molar conductivity, and elemental analyses . In all of the complexes, the ligand NFL was coordinated through two carboxyl oxygen atoms . Octahedral and tetrahegon geometries have been proposed for Fe(III)-, Co(II)-complexes and Zn(II)-complex, respectively . In vitro test of susceptibility of Fe(III)- and Zn(II)-complexes showed stronger activity than that of norfloxacin against G(-) E.Coli and Bacillus dysenteriae bacteria. J Appl Bacteriol, 1995 Oct, 79(4), 432 - 8 Biological indicators for low temperature steam and formaldehyde sterilization: the effect of defined media on sporulation, growth index and formaldehyde resistance of spores of Bacillus stearothermophilus strains; Wright AM et al.; Preliminary screening was carried out on spores of 29 strains of Bacillus stearothermophilus to determine their potential as biological indicator organisms for low temperature steam and formaldehyde sterilization . Each strain was sporulated on four chemically defined media . Fourteen strains produced satisfactory sporulation on one or more of the media but there was considerable variation in the extent of sporulation . The growth index of the spores, which was dependent on both the strain of organism and the sporulation medium, ranged from 1% to 90% . The spores were appraised on the basis of their resistance to inactivation by 0.5% w/v formaldehyde in aqueous solution at 70 degrees C . The survivor curves obtained could be characterized into five types on the basis of the shape of the curve . Only five strains of Bacillus stearothermophilus produced spores with the characteristics of high resistance, linear semi-logarithmic survivor curve and high growth index that would be required of a potential biological indicator organism. Radiology, 1995 Oct, 197(1), 67 - 72 Bacillary angiomatosis in patients with AIDS: multiorgan imaging findings; Moore EH et al.; PURPOSE: To describe the varied clinical manifestations and imaging findings encountered in bacillary angiomatosis, an infectious complication of the acquired immunodeficiency syndrome (AIDS) . MATERIALS AND METHODS: Clinical, imaging, and histopathologic findings in nine men (aged 26-50 years) with AIDS and bacillary angiomatosis were described . This condition often manifests as vascular skin lesions that resemble those of Kaposi sarcoma, fever, and anemia and is due to infection with Bartonella (Rochalimaea) henselae . RESULTS: Common imaging findings included lung nodules, mediastinal adenopathy, peripheral adenopathy, pleural effusions, ascites, abdominal adenopathy, soft-tissue masses, and low-attenuation lesions in the liver and/or spleen . Most notably, nodes and soft-tissue lesions were dramatically enhanced with injection of contrast material, which is presumably because the lesions are composed to a large extent of well-formed capillaries . CONCLUSION: Bacillary angiomatosis, a treatable infection, should be considered in patients with AIDS, particularly when Kaposi sarcoma is suspected clinically. J Infect Dis, 1995 Oct, 172(4), 1145 - 9 Prevalence of Bartonella henselae antibodies in pet cats throughout regions of North America; Jameson P et al.; Cat exposure has been directly associated with the development of human Bartonella henselae infections, resulting in cat-scratch disease, bacillary angiomatosis, or bacteremia . The prevalence of serum antibody titers to B . henselae was determined for selected pet cats from 33 geographic locations throughout the United States and several areas in western Canada . Seroprevalences paralleled increasing climatic warmth (P < .02) and annual precipitation (P < .03) . These warm, humid areas with the highest seroprevalence would also have the highest number of potential arthropod vectors . The southeastern United States, Hawaii, coastal California, the Pacific Northwest, and the south central plains had the highest average prevalences (54.6%, 47.4%, 40.0%, 34.3%, and 36.7%, respectively) . Alaska, the Rocky Mountain-Great Plains region, and the Midwest had low average prevalences (5.0%, 3.7%, and 6.7%, respectively) . Overall, 27.9% (175/628) of the cats tested were seropositive . The seroprevalence of B . henselae in cats varies throughout the United States and appears to be influenced by climate. Curr Microbiol, 1995 Oct, 31(4), 228 - 33 ssp genes and spore osmotolerance in Bacillus thuringiensis israelensis and Bacillus sphaericus; Cucchi A et al.; It was shown previously that spores and vegetative cells of Bacillus sphaericus (Bf) and Bacillus thuringiensis israelensis (Bti) are very sensitive to osmotic variations . Since spore osmotolerance has been associated with their SASP (small acid soluble spore proteins) content coded by ssp genes, hybridization assays were performed with sspE and sspA genes from B . subtilis as probes and showed that Bti and Bf strains could lack an sspE-like gene . The B . subtilis sspE gene was then introduced into Bti 4Q2 strain; spores were obtained and showed a 65 to 650 times higher level of osmotolerance to NaCl, without affecting other important properties: hypoosmotic resistance in vegetative cells, spore UV resistance, and larvicidal activity against diptera larvae. Nervenarzt, 1995 Oct, 66(10), 785 - 8 {Intracerebral hemorrhage and multiple brain abscesses caused by Bacillus cereus within the scope of acute lymphatic leukemia}; Strittmatter M et al.; Multiple hematogenic brain abscesses in immunosuppression are occasionally caused by rare and primary apathogenic causative agents . We report a first case of an isolated CNS infection by Bacillus cereus, which led to death from multiple brain abscesses and an intracerebral hemorrhage, probably caused by the infection, within 4 days . The underlying disease leading to immunosuppression was acute lymphatic leukemia in complete remission . In spite of antibiotic therapy the chemotherapy-induced neutropenia enabled unhindered spreading of the necrotizing infection, which was verified by histological analysis . The production of potent toxins such as hemolysin and cerelolysin by B . cereus leads to rapid and fulminant tissue destruction usually involving the walls of blood vessels. Appl Environ Microbiol, 1995 Oct, 61(10), 3592 - 7 Trehalose and sucrose protect both membranes and proteins in intact bacteria during drying; Leslie SB et al.; The microorganisms Escherichia coli DH5 alpha and Bacillus thuringiensis HD-1 show an increased tolerance to freeze-drying when dried in the presence of the disaccharides trehalose and sucrose . When the bacteria were dried with 100 mM trehalose, 70% of the E . coli and 57% of the B . thuringiensis organisms survived, compared with 56 and 44%, respectively, when they were dried with sucrose . Only 8% of the E . coli and 14% of the B . thuringiensis organisms survived drying without the sugars . Fourier transform infrared spectroscopy was used to investigate the role of membrane phase transitions in the survival of the organisms during drying and rehydration . Both E . coli and B . thuringiensis showed an increase of 30 to 40 degrees C in the temperature of their phospholipid phase transition when dried without the sugars, while phase transition temperatures of those dried with the sugars remained near those of the hydrated cells . A Fourier transform infrared spectroscopy microscope made it possible to investigate the effects of drying on the protein structure in the intact cells . The amide II peak shifts from 1,543 cm-1 in the hydrated cells to about 1,533 cm-1 in the cells dried without sugar . There is no shift in the amide II peak when the cells are dried with trehalose or sucrose . We attribute the increased survival to the sugars' ability to lower the membrane phase transition temperature and to protect protein structure in the dry state.(ABSTRACT TRUNCATED AT 250 WORDS) South Med J, 1995 Oct, 88(10), 1025 - 30 Tuberculosis screening in adults who have received bacille Calmette-Guérin vaccine; Ballew KA et al.; The incidence of tuberculosis among immigrants to the United States is 12 times that in the native-born population . Screening immigrants for tuberculosis is complicated by the widespread use of bacille Calmette-Guerin (BCG) vaccination . To determine the utility of tuberculin testing in adults who have been vaccinated with BCG vaccine, we studied the tuberculin reactions of 80 adults who came to us for naturalization physical examinations . No adverse effects were reported from tuberculin testing . Subjects from regions with a low prevalence of tuberculosis who had received BCG vaccine were significantly more likely to have a positive reaction than subjects who had not received BCG vaccine . However, among subjects from regions with a high prevalence of tuberculosis, there was no difference in the prevalence of positive reactions between those who reported having BCG vaccinations and those who said they had not . Interpretation of tuberculin reactions in immigrants who receive BCG vaccinations depends on the prevalence of tuberculosis in the country of origin . Adults receiving BCG vaccination who have a positive reaction and no evidence of active tuberculosis should receive prophylactic therapy or be observed carefully. Biochim Biophys Acta, 1995 Sep 27, 1252(1), 35 - 42 Comparison of primary structures and substrate specificities of two pullulan-hydrolyzing alpha-amylases, TVA I and TVA II, from Thermoactinomyces vulgaris R-47; Tonozuka T et al.; Thermoactinomyces vulgaris R-47 produces two alpha-amylases, TVA I, an extracellular enzyme, and TVA II, an intracellular enzyme . Both enzymes hydrolyze pullulan to produce panose, and also hydrolyze cyclodextrins . We cloned and sequenced the TVA I gene . The TVA I gene consisted of 1833 base pairs, and the deduced primary structure was composed of 611 amino-acid residues, including an N-terminal signal sequence consisting of 29 amino-acid residues . The similarity between the amino-acid sequence of mature TVA I with those of other pullulan/cyclodextrin-hydrolyzing enzymes, such as TVA II and Bacillus stearothermophilus neopullulanase, was only 30%, although that of TVA II with neopullulanase was 48% . TVA II prefers specific small oligosaccharides and alpha- and beta-cyclodextrins . Whereas kcat/Km values of TVA I for pullulan were larger than that of TVA II, and TVA II could not hydrolyze starch completely . TVA II was inhibited by maltose, the hydrolysate of starch, which seems to be the reason for inefficient hydrolysis of starch . These kinetic properties indicate that TVA I and TVA II have differential physiological roles in sugar metabolism extracellularly and intracellularly, respectively. Biochemistry, 1995 Sep 26, 34(38), 12302 - 10 Probing enzymic transition state hydrophobicities; Wangikar PP et al.; Hydrophobic interactions are important in numerous biological processes; however, the nature and extent of hydrophobic interactions in nonaqueous enzymology remain poorly defined . We have estimated the free energies of enzyme--substrate hydrophobic interactions for a model reaction catalyzed by subtilisin BPN'(from Bacillus amyloliquefaciens) in various solvents . Transition state stabilization of subtilisin in water has contributions from both ground state destabilization of hydrophobic substrates and intrinsic enzyme--substrate hydrophobic interactions . Both contributions are evident even in hydrophobic organic solvents and can be modified by protein engineering of the enzyme's binding site, as well as by changing the hydrophobicity of the reaction medium . We have also developed a method to estimate the hydrophobicity of the enzymic transition state involving systematic variation of the substrate and solvent hydrophobicities . The observed binding pocket hydrophobicities were directly affected by replacing the Gly166 residue, located at the back of hydrophobic S1 binding pocket of subtilisin BPN', with more hydrophobic amino acids such as alanine and valine . Thus, the observed S1 binding pocket hydrophobicities of the wild-type, G166A, and G166V mutants were measured to be 1.2, 1.8, and 2.6 log P units, respectively . Our method of calculating effective binding pocket hydrophobicity was found to be applicable to other enzymes, including horseradish peroxidase and alpha-chymotrypsin . Measurements of the binding pocket hydrophobicities have significant implications toward tailoring enzyme function in aqueous as well as nonaqueous media. Gene, 1995 Sep 22, 163(1), 65 - 8 Cloning and complete sequence of the DNA polymerase-encoding gene (BstpolI) and characterisation of the Klenow-like fragment from Bacillus stearothermophilus; Phang SM et al.; A fragment of the DNA polymerase I-encoding gene (polI) from Bacillus stearothermophilus (Bst) was obtained by PCR . This was used as a probe to obtain a full-length gene from a Bst genomic DNA (gDNA) plasmid library . Comparison of the sequence to B . caldotenax (Bca) showed about 93% homology at the amino acid (aa) level . A Klenow-like (BstpolIk) clone was developed and the recombinant protein displayed DNA polymerase activity similar to the wild-type BstPolI enzyme. Schweiz Med Wochenschr, 1995 Sep 16, 125(37), 1715 - 25 {Ludwig Lichtheim and Robert Koch . How the discovery of the tubercle bacillus in Switzerland was made known (1882/1883)}; Boschung U et al.; The discovery of the tubercle bacillus by Robert Koch in March 1882 was immediately reported by the medical press, including that of Switzerland . Among the clinicians most active in introducing the new views and techniques to Swiss medical practitioners was Ludwig Lichtheim (1845-1928), professor of internal medicine in Berne (1879-1888), who had been acquainted with Koch since their time in Breslau . Lichtheim's successful efforts are illustrated by quotations from his (unpublished) memoirs. Eur J Biochem, 1995 Sep 15, 232(3), 849 - 58 Crystal and molecular structure at 0.16-nm resolution of the hybrid Bacillus endo-1,3-1,4-beta-D-glucan 4-glucanohydrolase H(A16-M); Hahn M et al.; H(A16-M) is a hybrid endo-1,3-1,4-beta-D-glucan 4-glucanohydrolase from Bacillus . Its crystal structure was refined using synchrotron X-ray diffraction data up to a maximal resolution of 0.16 nm . The R value of the resulting model is 14.3% against 21,032 reflections > 2 sigma . 93% of the amino acid residues are in the most favorable regions of the Ramachandran diagram, and geometrical parameters are in accordance with other proteins solved at high resolution . As shown earlier {Keitel, T., Simon, O., Borriss, R . & Heinemann, U . (1993) Proc . Natl Acad . Sci . USA 90, 5287-5291}, the protein folds into a compact jellyroll-type beta-sheet structure . A systematic analysis of the secondary structure reveals the presence of two major antiparallel beta-sheets and a three-stranded minor mixed sheet . Amino acid residues involved in catalysis and substrate binding are located inside a deep channel spanning the surface of the protein . To investigate the stereochemical cause of the observed specificity of endo-1,3-1,4-beta-D-glucan 4-glucanohydrolases towards beta-1,4 glycosyl bonds adjacent to beta-1,3 bonds, the high-resolution crystal structure has been used to model an enzyme-substrate complex . It is proposed that productive substrate binding to the subsites p1, p2 and p3 of H(A16-M) requires a beta-1,3 linkage between glucose units bound to p1 and p2. Eur J Biochem, 1995 Sep 15, 232(3), 839 - 43 Abnormally high pKa of an active-site glutamic acid residue in Bacillus circulans xylanase . The role of electrostatic interactions; Davoodi J et al.; The active site of Bacillus circulans xylanase (1,4-beta-D-xylanohydrolase, EC 3.2.1.8) contains two glutamic acid residues, Glu78 and Glu172, which are crucial for the catalytic activity of the enzyme . Fourier-transform infrared spectroscopy was used to determine the ionization state of these residues as a function of pH . For the wild-type enzyme, titration of one of the carboxylate groups occurs at pH 6.8 . This titration is absent in the Glu78-->Gln and Glu172-->Gln variants of the enzyme . This, together with crystallographic data, indicates that Glu172 has an abnormally high pKa of 6.8, caused largely by electrostatic interactions of this residue with the proximal Glu78 . Differential scanning calorimetry experiments with the wild-type xylanase and a number of its mutants have shown that the presence of two nearby carboxyl groups results in a pH-dependent destabilization of the protein structure. Carbohydr Res, 1995 Sep 15, 275(1), 147 - 54 Structure of the capsular polysaccharide from Alteromonas sp . CMM 155; Zubkov VA et al.; Capsular polysaccharide (CPS) was obtained by water-saline extraction of the Alteromonas sp . CMM 155 . On the basis of solvolysis with anhydrous HF and 1H- and 13C-NMR spectral data, including NOE experiments, it was concluded that the capsular polysaccharide had the following structure containing novel N-acyl-amino sugar and bacillosamine residues: --> 3)-alpha-D-GalpNAc-(1 --> 4)-alpha-L-GalApNAc(1 --> 3)- alpha-D-QuipNAc4NAc-(1 --> 3)-beta-D-Quip4NAlaAc-(1 --> Biochem J, 1995 Sep 15, 310 ( Pt 3), 939 - 43 Effect of replacement of ferriprotoporphyrin IX in the haem domain of cytochrome P-450 BM-3 on substrate binding and catalytic activity; Modi S et al.; Bacillus megaterium cytochrome P-450 BM-3 (coded by gene CYP102) is a catalytically self-sufficient mono-oxygenase, with both cytochrome P-450 and NADPH:cytochrome P-450 reductase domains, that catalyses the hydroxylation of fatty acids . The natural ferriprotoporphyrin IX has been removed from the haem domain of cytochrome P-450 BM-3 by treatment with acidified acetone, and it has been shown that, under carefully controlled conditions, haem can be added back to the resultant apoprotein to obtain a fully reconstituted haem domain with spectroscopic, substrate-binding and catalytic properties indistinguishable from those of the native domain . Replacement of the natural haem with ferriprotoporphyrin IX dimethyl ester yields a protein which has a higher affinity for the substrate dodecanoic acid and (in the presence of the reductase domain) the same catalytic rate as the native haem domain . Replacement with ferrimesoporphyrin IX yields a protein with the same affinity for substrate, but a reduced catalytic turnover . These results suggest that the haem moiety has a role in the creation of the binding pocket for substrate, and that modification of the electron density on the haem iron effects the catalytic rate. EMBO J, 1995 Sep 15, 14(18), 4578 - 88 Protein-rRNA binding features and their structural and functional implications in ribosomes as determined by cross-linking studies; Urlaub H et al.; We have investigated protein-rRNA cross-links formed in 30S and 50S ribosomal subunits of Escherichia coli and Bacillus stearothermophilus at the molecular level using UV and 2-iminothiolane as cross-linking agents . We identified amino acids cross-linked to rRNA for 13 ribosomal proteins from these organisms, namely derived from S3, S4, S7, S14, S17, L2, L4, L6, L14, L27, L28, L29 and L36 . Several other peptide stretches cross-linked to rRNA have been sequenced in which no direct cross-linked amino acid could be detected . The cross-linked amino acids are positioned within loop domains carrying RNA binding features such as conserved basic and aromatic residues . One of the cross-linked peptides in ribosomal protein S3 shows a common primary sequence motif--the KH motif--directly involved in interaction with rRNA, and the cross-linked amino acid in ribosomal protein L36 lies within the zinc finger-like motif of this protein . The cross-linked amino acids in ribosomal proteins S17 and L6 prove the proposed RNA interacting site derived from three-dimensional models . A comparison of our structural data with mutations in ribosomal proteins that lead to antibiotic resistance, and with those from protein-antibiotic cross-linking experiments, reveals functional implications for ribosomal proteins that interact with rRNA. Biochem Biophys Res Commun, 1995 Sep 14, 214(2), 602 - 7 Expression and synergistic effect of three types of crystal protein genes in Bacillus thuringiensis; Park HW et al.; Bacillus thuringiensis NT0423 newly isolated from sericultural farms in Korea produces quite atypical bipyramidal crystals of a common major band of ca . 130 kDa, and has dual specificity against Lepidoptera and Diptera . To enforce the Diptera toxicity of B . thuringiensis NT0423, cryIVD and cytA genes were transformed into B . thuringiensis NT0423 . The transformant B . thuringiensis PT0529 was obtained from introduction of pCG5 into B . thuringiensis NT0423 by electroporation . The expression of crystal proteins in B . thuringiensis PT0529 was characterized by SDS-PAGE and electron microscopy . The results showed that crystalline inclusions of host, CryIVD and CytA were clearly observed in B . thuringiensis PT0529 . Furthermore, the toxicity of B . thuringiensis PT0529 against Diptera was highly enforced by synergistic effect. Biochim Biophys Acta, 1995 Sep 14, 1258(2), 122 - 34 Relation between micellar structure of model bile and activity of esterase; Uematsu S et al.; In a model bile solution composed of lecithin (L)-bile salt (B), the solubilization of lipid and the accessibility of enzyme to the lipid were examined by observation of EPR spectra and measurement of enzyme activity . The lifetime of the spin probe in the micellar phase was estimated to be approx . 1 microsecond by means of line shape analysis . Both population and lifetime increased with temperature and the molar ratio of lecithin to bile salt (L/B) . The EPR data indicated that simple micelle of bile salt, mixed disk micelle of bile salt-lecithin, and multi-lamellar mixed disk micelle can exist in a model bile solution, depending on the L/B molar ratio across a range from 0 to 1.5 . The maximal power of the mixed disk micelle to solubilize cholesteryl ester in the model bile at a L/B molar ratio of 1:1 was confirmed by EPR measurement of cholesteryl 12-DOXYL-stearate . Observation of the enzyme activity on a mixture of model bile and substrate at 37 degrees C revealed selective accessibility of cholesterol esterase (bovine pancreas) to mixed disk micelle, of cholesterol oxidase (Streptomyces cinnamomeus) to both simple and mixed disk micelle, and of pancreatic lipase (porcine pancreas) to both simple micelle and an oil droplet of substrate . The temperature-dependent activity of cholesterol oxidase to cholesterol in mixed disk micelle can be explained in terms of mesomorphic phase transition of lecithin side chains followed with fluidity of liquid crystal phase . Regarding phospholipase C from Bacillus cereus, though the selective accessibility to the micelles was not observed at 37 degrees C, a decrease in activity for mixed disk micelle could be found at lower temperatures. Proc Natl Acad Sci U S A, 1995 Sep 12, 92(19), 8768 - 72 The urease locus of Mycobacterium tuberculosis and its utilization for the demonstration of allelic exchange in Mycobacterium bovis bacillus Calmette-Guérin; Reyrat JM et al.; The ureABC genes of Mycobacterium tuberculosis were cloned . By using a set of degenerate primers corresponding to a conserved region of the urease enzyme (EC 3.5.1.5), a fragment of the expected size was amplified by PCR and was used to screen a M . tuberculosis cosmid library . Three open reading frames with extensive similarity to the urease genes from other organisms were found . The locus was mapped on the chromosome, using an ordered M . tuberculosis cosmid library . A suicide vector containing a ureC gene disrupted by a kanamycin marker (aph) was used to construct a urease-negative Mycobacterium bovis bacillus Calmette-Guerin mutant by allelic exchange involving replacement of the ureC gene with the aph::ureC construct . To our knowledge, allelic exchange has not been reported previously in the slow-growing mycobacteria . Homologous recombination will be an invaluable genetic tool for deciphering the mechanisms of tuberculosis pathogenesis, a disease that causes 3 x 10(6) deaths a year worldwide. Arch Biochem Biophys, 1995 Sep 10, 322(1), 43 - 52 Stringency of substrate specificity of Escherichia coli malate dehydrogenase; Boernke WE et al.; Malate dehydrogenase and lactate dehydrogenase are members of the structurally and functionally homologous family of 2-ketoacid dehydrogenases . Both enzymes display high specificity for their respective keto substrates, oxaloacetate and pyruvate . Closer analysis of their specificity, however, reveals that the specificity of malate dehydrogenase is much stricter and less malleable than that of lactate dehydrogenase . Site-specific mutagenesis of the two enzymes in an attempt to reverse their specificity has met with contrary results . Conversion of a specific active-site glutamine to arginine in lactate dehydrogenase from Bacillus stearothermophilus generated an enzyme that displayed activity toward oxaloacetate equal to that of the native enzyme toward pyruvate (H . M . Wilks et al . (1988) Science 242, 1541-1544) . We have constructed a series of mutants in the mobile, active site loop of the Escherichia coli malate dehydrogenase that incorporate the complementary change, conversion of arginine 81 to glutamine, to evaluate the role of charge distribution and conformational flexibility within this loop in defining the substrate specificity of these enzymes . Mutants incorporating the change R81Q all had reversed specificity, displaying much higher activity toward pyruvate than to the natural substrate, oxaloacetate . In contrast to the mutated lactate dehydrogenase, these reversed-specificity mutants were much less active than the native enzyme . Secondary mutations within the loop of the E . coli enzyme (A80N, A80P, A80P/M85E/D86T) had either no or only moderately beneficial effects on the activity of the mutant enzyme toward pyruvate . The mutation A80P, which can be expected to reduce the overall flexibility of the loop, modestly improved activity toward pyruvate . The possible physiological relevance of the stringent specificity of malate dehydrogenase was investigated . In normal strains of E . coli, fermentative metabolism was not affected by expression of the mutant malate dehydrogenase . However, when expressed in a strain of E . coli unable to ferment glucose, the mutant enzyme restored growth and produced lactic acid as the sole fermentation product. J Biol Chem, 1995 Sep 8, 270(36), 21299 - 306 Evidence for a bifurcation of the mitogenic signaling pathway activated by Ras and phosphatidylcholine-hydrolyzing phospholipase C; Bjorkoy G et al.; NIH 3T3 cells stably transfected with the gene encoding phosphatidylcholine-hydrolyzing phospholipase C (PC-PLC) from Bacillus cereus display a chronic elevation of intracellular diacylglycerol levels and a transformed phenotype . We have used such PC-PLC-transformed cells to evaluate the roles of the cytoplasmic serine/threonine kinases Raf-1, zeta protein kinase C (zeta PKC) and protein kinase A (PKA) in oncogenesis and mitogenic signal transduction elicited by phosphatidylcholine hydrolysis . We demonstrate here that stable expression of dominant negative mutants of both zeta PKC and Raf-1 lead to reversion of PC-PLC-transformed cells . Interestingly, expression of kinase defective zeta PKC also reverted NIH 3T3 cells transformed by the v-Ha-ras oncogene . Activation of PKA in response to elevation of cAMP levels also lead to reversion of PC-PLC-induced transformation, implicating PKA as a negative regulator acting downstream of PC-PLC . On the other hand, inhibition or depletion of phorbol ester responsive PKCs attenuated but did not block the ability of PC-PLC-transformed cells to induce DNA synthesis in the absence of growth factors . These results clearly implicate both Raf-1 and zeta PKC as necessary downstream components for transduction of the mitogenic/oncogenic signal generated by PLC-mediated hydrolysis of phosphatidylcholine and suggest, together with other recent evidence, a bifurcation in the signaling pathway downstream of PC-PLC. Ann Soc Belg Med Trop, 1995 Sep, 75(3), 201 - 10 {Epidemic of bacillary dysentery in the Rwanda refugee camps of the Goma region (Zaire, North Kivu) in August 1994}; Milleliri JM et al.; The authors describe the extension of an outbreak of bacillary dysentery among the Rwandese populations seeking refuge in the region of Goma, Zaire in august 1994 . Analysis of the epidemiological surveillance data and of the bacteriological laboratory results of the Bioforce, show that this epidemic was probably facilitated by the preceding cholera outbreak . In such circumstances, rapid sterilization of the virus reservoir, by short course treatments, might be beneficial in limiting the extension of the epidemic. Protein Eng, 1995 Sep, 8(9), 939 - 45 Contribution of a disulfide bridge to the stability of 1,3-1,4-beta-D-glucan 4-glucanohydrolase from Bacillus licheniformis; Pons J et al.; Bacillus 1,3-1,4-beta-glucanases possess a highly conserved disulfide bridge connecting a beta-strand with a solvent-exposed loop lying on top of the extended binding site cleft . The contribution of the disulfide bond and of both individual cysteines (Cys61 and Cys90) in the Bacillus licheniformis enzyme to stability and activity has been evaluated by protein engineering methods . Reduction of the disulfide bond has no effect on kinetic parameters, has only a minor effect on the activity-temperature profile at high temperatures, and destabilizes the protein by less than 0.7 kcal/mol as measured by equilibrium urea denaturation at 37 degrees C . Replacing either of the Cys residues with Ala destabilizes the protein and lowers the specific activity . C90A retains 70% of wild-type (wt) activity (in terms of Vmax), whereas C61A and the double mutant C61A-C90A have 10% of wt Vmax . A larger change in free energy of unfolding is seen by equilibrium urea denaturation for the C61A mutation (loop residue, 3.2 kcal/mol relative to reduced wt) as compared with the C90A mutation (beta-strand residue, 1.8 kcal/mol relative to reduced wt), while the double mutant C61A-C90A is approximately 0.8 kcal/mol less stable than the single C61A mutant . The effects on stability are interpreted as a result of the change in hydrophobic packing that occurs upon removal of the sulfur atoms in the Cys to Ala mutations. J Appl Toxicol, 1995 Sep-Oct, 15(5), 365 - 73 Semiautomated quantification of cytotoxic damage induced in cultured insect cells exposed to commercial Bacillus thuringiensis biopesticides; Tayabali AF et al.; A convenient in vitro bioassay based on semiautomated quantification of live-cell reduction of tetrazolium dyes--3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and 2,3-bis(2-methoxy-4-nitro-5-sulphophenyl)-2H-tetrazolium-5-corb oxanilide sodium salt (XTT)--to formazan was developed and used to evaluate cytotoxic effects of two commercial insecticides (BT) derived from Bacillus thuringiensis subsp . kurstaki (Btk) . Comparison of two target insect cell lines MG1 (Trichoplusia ni, cabbage looper midgut) and Sf9 (Spodoptera frugiperda, fall army worm oocyte) revealed similar cell-dependent responses in mitochondrial-associated electron transport activity . The 50% inhibition of formazan production (ID50) obtained by exposing these cells to 20 microM 2,4-dinitrophenol or 5 mM sodium azide occurred in the range 10(-7)-10(-6) International Units (IU) of BT cell(-1) 24 h(-1) . Damage to cell adhesion and cytoarchitecture, revealed by light and electron microscopic analysis, increased with BT exposure and dose . MTT was superior to XTT as a cytotoxic indicator in kinetic studies related to spores, a major component of BT . Unless blocked by antibiotic (gentamicin), vegetative growth resulting from spore germination was the major cause of toxicity . The ID50 exposure time using vegetative Btk cells was approximately 0.1-0.2 times that required for BT spores, with or without intact parasporal proteins present . This difference in exposure is an indirect measure of the time required for spores to germinate and produce vegetative cells . The assay methodology developed here, if linked with suitable target and non-target animal cell types, should have broad application for conducting standardizable estimates of cytotoxic potential of any microbe-based biotechnology product. Antibiot Khimioter, 1995 Sep, 40(9), 17 - 9 {Preparation and antiviral properties of Bacillus intermedius ribonuclease modified by chloranhydride of adamantanecarboxylic acid}; Kurinenko BM et al.; Bacillus intermedius ribonuclease modified by the residue of adamantane carboxylic acid was prepared . When the cells of chick embryo fibroblasts infected by the fowl plague virus were exposed to the modified ribonuclease, the antiviral activity proved to be higher by comparison to that of the native enzyme . The chemotherapeutic index of the RNAse after the modification increased 4 times. Eur Respir J Suppl, 1995 Sep, 20, 633s - 648s Mobile genetic elements in mycobacteria; Dale JW; Mobile genetic elements, ranging from plasmids and bacteriophages to insertion sequences and transposons, have come to play key roles in many aspects of basic and applied research in mycobacteriology . Plasmids and bacteriophages have been widely used as cloning vectors, especially for constructing recombinant vaccines based on bacille Calmette-Guerin (BCG); composite transposons have also been used for this purpose . At the same time, insertion sequences have proved invaluable for diagnostic and epidemiological studies, and transposon mutagenesis provides a useful method for inactivating and marking selected mycobacterial genes . Plasmids are commonly found in many mycobacterial species, notably M . avium, although not in M . tuberculosis; the biological significance of these plasmids (if any) is mostly unknown . Insertion sequences, and other repetitive elements, have also been characterized from many mycobacterial species . Special attention is paid to IS6110/IS986, from M . tuberculosis, and the IS900 family from M . avium and related organisms; the latter includes the recently described highly mobile element IS1110 . The emphasis of the paper is on the molecular biology and significance of plasmids and insertion sequences/transposons, in mycobacteria and in bacteria of plasmids and insertion sequences/transposons, in mycobacteria and in bacteria in general, and their applications as cloning vectors and in transposon mutagenesis. J Am Mosq Control Assoc, 1995 Sep, 11(3), 329 - 34 Role of larval cadavers in recycling processes of Bacillus sphaericus; Becker N et al.; The influence of larval cadavers of Culex pipiens on recycling processes of Bacillus sphaericus was investigated by bioassays and spore counts in the laboratory . Studies conducted with 3 different B . sphaericus concentrations (0.005, 0.01, 0.05 mg B . sphaericus/liter) indicated that the presence of cadavers in the water contributed to the maintenance of toxic levels of B . sphaericus . Larval cadavers seem to contain all the nutrients necessary both for vegetative multiplication and for toxin synthesis associated with the sporulation process . Bioassays of B . sphaericus revealed that the mortality of Culex pipiens remained on a high level over a period of 26 days when larval cadavers were added every second day to the test vessels . This result was supported by a sharp increase in spore density when cadavers were added at the same interval . The test series showed B . sphaericus recycles in intact cadavers of Culex pipiens, whereas this phenomenon could not be observed when crushed cadavers were used in the trials . Therefore, our results demonstrated that for successful recycling processes it seems of crucial importance that infected cadavers remain intact at least for a certain time and also that the dosage of the applied B . sphaericus plays a major role in recycling processes whereas larval density is only of minor importance to these processes. Acta Trop, 1995 Sep, 60(1), 47 - 57 The potential for malaria control with the biological larvicide Bacillus thuringiensis israelensis (Bti) in Peru and Ecuador; Kroeger A et al.; A study on the efficacy of Bti spraying in mosquito breeding places was undertaken in the Pacific coast of Peru and Ecuador and in the Amazon area of Peru . It was shown that Bti is a powerful larvicide for Anopheles larvae, although it sinks quickly, whereas Anopheles larvae feed at the water surface . The duration of its effect was less than 7 days with the exception of the Amazon area of Peru, where it was approximately 10 days . In two study areas, Bti was sprayed weekly over periods of 10 and 7 weeks, respectively, and the adult mosquito densities were monitored . The Anopheles adult density (bites per person per hour on human baits) was reduced by an average of 70% in one area and by up to 50% in the other . This means that Bti spraying can potentially be an important component of a modified malaria control strategy. J Microencapsul, 1995 Sep-Oct, 12(5), 515 - 23 Control of mosquito larvae by encapsulated pathogen Bacillus thuringiensis var . israelensis; Elcin YM; Bacillus thuringiensis var . israelensis (B.t.i.) containing alginate microcapsules were prepared in order to maintain durable formulations which could resist several effects causing reduced efficiency during applications . B.t.i . spores were harvested through NYSM agar plates and encapsulated in Ca-alginate (0.5-2.0% w/v) gels without any significant loss of sporal or larvicidal activity . The effect of acidic pH on the larvicidal toxin was tested using Culex sp . larvae in the laboratory . The alginate microcapsules pretreated with saturated KH2PO4 solution gave larvicidal activity after 24-48 h, by bioassay . Suspension and encapsulated forms of the pathogenic bacterium were exposed to pH variations (3.0-10.0), UV light and high temperature (50 degrees C) . Durability to Pb++, Cu++, Fe++ compounds and phenol was also examined . As the alginate content increased, stability of B.t.i . drastically increased against the tested effects, but to obtain useful releasing microcapsules, 1.0-1.5% w/v alginate concentrations were found to be optimum. Mikrobiologiia, 1995 Sep-Oct, 64(5), 623 - 9 {Biosynthesis of alkaline extracellular proteinases in Bacillus intermedius}; Itskovich EL et al.; The conditions for growth and serine proteinase biosynthesis were studied in the batch culture of Bacillus intermedius . The synthesis of the enzyme was found to be inhibited by glucose and by the amino acid mixture and activated by the addition of organic compounds-casein and gelatin-to the medium . Inorganic phosphate in the medium increased the enzyme yield . The enzyme activity was shown to be twofold higher upon the addition of corn extract (1.8%) in the medium than in the medium with meat peptone (1.7-2.2%) and inorganic phosphate (0.28-0.30 miligram). Mikrobiologiia, 1995 Sep-Oct, 64(5), 596 - 600 {Effect of membranotropic physiologically-active substances on the growth of Saccharomyces cerevisiae cultures}; Kupriianova-Ashina FG et al.; The effect of Bacillus intermedius RNase and yeast autoregulatory d2 factor on growth of Saccharomyces cerevisiae is reported . It is shown that 0.01 microgram/ml of RNase stimulates the growth of the yeast on malt wort and molasses, the peak of the effect being observed for the enzyme added in the middle exponential phase . Certain concentrations of the d2 factor can also potentiate yeast growth . The effect is more pronounced if the d2 factor and RNase are added in combination rather than singly . Although the cell membrane is a target for both d2 factor and RNase, the mechanisms of their action are presumably different. Yakugaku Zasshi, 1995 Sep, 115(9), 756 - 64 {Study on surfactin, a cyclic depsipeptide . I . Isolation and structure of eight surfuctin analogs produced by Bacillus natto KMD 2311}; Kanatomo S et al.; Crude surfactin was simply prepared from the culture filtrate of Bacillus natto KMD 2311 twice by acidification of the filtrate and extraction of the precipitate with ethanol . Eight surfactin analogs were isolated from the crude surfactin by RP-HPLC and gel filtration . The structure of each analog was deduced by means of amino acid composition of the acid hydrolysate and FAB-MS measurement to be a cyclic depsipeptide containing a hydroxyfatty acid . The structure of the hydroxyfatty acid moieties was elucidated as n- or iso- or anteiso-3-hydroxyfatty acid composed of carbon number 13-16 by GC analysis and EI-MS after the methanolysis of the analogs . The amino acid sequence of the peptide portion was assigned as acyl-Glu-Leu-Leu-Val-Asp-Leu-Leu by EI-MS for eight analogs . The isolated four compounds were found to be identical with the known surfactin analogs, A1, B1, B2 and C1 . Although surfactin A2 and C2 had not been isolated, their structures were deduced to be a surfactin analog . Surfactin A3 and D were novel analogs . The acyl groups of surfactin A2, A3, C2 and D were anteiso-3-hydroxytridecanoic acid, n-3-hydroxytridecanoic acid, anteiso-3-hydroxypentadecanoic acid and iso-3-hydroxyhexadecanoic acid, respectively. Yakugaku Zasshi, 1995 Sep, 115(9), 742 - 8 {Examination of toxin production from environmental Bacillus cereus and Bacillus thuringiensis}; Mikami T et al.; Emetic toxin- and entero toxin-producing activities were examined for 524 strains of Bacillus cereus and 90 strains of Bacillus thuringiensis to determine the distribution and contamination in natural environment, including foods, vegetable, soil and etc . Emetic toxin was assayed by the production of acid from HEp-2 cells induced by the culture supernatants of Bacillus, which was developed as an improved method for the detection of B . cereus heat-stable emetic-toxin by us . In the 524 strains of B . cereus tested, 11 out of 60 strains obtained from bean paste and 5 out of 20 strains obtained from peach produced emetic toxin which was detected by the HEp-2 cell method . All these emetic toxin-producing isolate strains were classified into H1 serotype . The 90 strains of B . thuringiensis did not produce emetic toxin, but the 47 strains produced entero toxin. Bull Pan Am Health Organ, 1995 Sep, 29(3), 226 - 36 Restriction fragment length polymorphism (RFLP) analysis and tuberculosis epidemiology; Gomez Marin JE et al.; In order to study polymorphisms of the DNA insertion sequence 6110 (IS6110) in Mycobacterium tuberculosis strains isolated from Colombian patients, together with resistance to antituberculous medications in the Department of Quindio, Colombia, a prospective study was conducted using a consecutive sample of 59 patients with symptomatic pulmonary tuberculosis whose cases had been confirmed by bacilloscopy, both with and without a history of treatment . The patients, who were participating in the Tuberculosis Control Program of the Regional Health Institute of Quindio in Armenia, included all individuals attending local health centers and hospitals between March and July 1993 who were referred to the regional institute . Sputum specimens from each patient were cultured and subjected to drug sensitivity tests . Subsequently, restriction fragment length polymorphisms (RFLP) of IS6110 from 27 patients were analyzed . The patients' treatment histories were used to classify their cases according to WHO criteria . Forty-five cultures were found positive, 44 for M . tuberculosis and 1 for M . africanum . Initial drug resistance was observed in 4 of 42 new cases, or 9.5% (95% CI: 0.6, 18), 2 showing resistance to isoniazid (INH) and 2 to isoniazid plus streptomycin (INH-SM) . Acquired resistance was observed in 2 of the 3 chronic cases and relapses, the bacteria being resistant to isoniazid, rifampicin, and streptomycin (INH-RM-SM) in one case and to isoniazid, ethambutol, rifampicin, and streptomycin (INH-EMB-RM-SM) in the other . In those 27 strains subjected to RFLP analysis, the number of copies of IS6110 ranged from 6 to 17 . Similarity coefficients revealed five distinct groups of strains . Overall, the RFLP analysis permitted most of the strains to be distinguished from one another, implying that the polymorphisms involved are sufficient to permit effective employment of this technique, which appears to have considerable potential for use in epidemiologic studies and in work designed to provide a basis for tuberculosis control program decision-making. Am J Respir Crit Care Med, 1995 Sep, 152(3), 1041 - 9 BCG priming enhances endotoxin-induced acute lung injury independent of neutrophils; Tasaka S et al.; Bacillus Calmette Guerin (BCG) is known to increase susceptibility to endotoxin in some animal species . We investigated the effect of BCG-priming and the role of neutrophils in the priming process on the pathogenesis of acute lung injury caused by intravenously administered Escherichia coli endotoxin (LPS) . Guinea pigs were divided into seven groups: (1) control (n = 8), (2) BCG-alone (n = 6), (3) cyclophosphamide (CPA)-alone (n = 6), (4) CPA+LPS (n = 6), (5) LPS-alone (n = 6), (6) BCG+LPS (n = 6), and (7) BCG+CPA+LPS (n = 6) . A BCG dose of 8 mg/kg was injected subcutaneously 10 d before the study . CPA was administered intraperitoneally to induce peripheral neutropenia . Animals were observed for 4 h after intravenous administration of 0.2 mg/kg of LPS . The plasma TNF level was measured 2 h after LPS challenge . Lung wet-to-dry weight ratio, {125I} albumin leakage in lung tissue, differential cell count in bronchoalveolar lavage (BAL) fluid, and histopathologic features were examined immediately after death . Although the LPS-alone group showed PMN accumulation in lung tissue, neither excess lung water nor increased albumin leakage was induced by this dose of LPS . The BCG+LPS group showed increased lung water, histopathologic edema, and increases in BAL fluid cell counts and plasma TNF in comparison with the LPS-alone group . The BCG+CPA+LPS group also showed enhanced lung injury comparable to that seen in the BCG+LPS group . In both the CPA-alone and the CPA+LPS groups, no parameter was increased as compared with those in the control group.(ABSTRACT TRUNCATED AT 250 WORDS) Ann Emerg Med, 1995 Sep, 26(3), 290 - 5 Delayed recognition and infection control for tuberculosis patients in the emergency department; Moran GJ et al.; STUDY OBJECTIVE: The recent increase in tuberculosis (TB) cases may have an important effect on emergency department infection-control measures . We describe infection-control interventions for TB patients admitted through the ED and hypothesize that ED suspicion of TB is associated with more rapid isolation and treatment . DESIGN: Retrospective chart review . SETTING: The ED of a 400-bed urban, university-affiliated county hospital . PARTICIPANTS: Fifty-five patients with TB culture-positive and acid-fast bacillus stain-positive respiratory specimens who were evaluated in the ED during 1991 and 1992 . RESULTS: We identified cases from the mycobacteriology log . Demographic and historical data and time elapsed before initiation of infection-control measures and TB therapy were recorded . We assessed the relationships of individual clinical findings and the ED presumptive diagnosis of TB (predictor variables) to time elapsed before isolation and therapy (outcome variables) with the log-rank test . The median time (interquartile range) from ED registration to isolation was 8 hours (range, 3 to 13 hours) . An ED presumptive diagnosis of TB was made in 71% of cases and was significantly associated with shorter time elapsed before isolation (5 hours {range, 2 to 10 hours} versus 21 hours {range, 11 to 111 hours}; P < .001) and less time elapsed before therapy (12 hours {range, 9 to 22 hours} versus 128 hours {68 to 374 hours}; P < .001) . We found TB exposure, radiographic changes typical of TB, absence of HIV risk factors, presence of cough, and sputum production to be associated with more rapid isolation . CONCLUSION: Among patients with active pulmonary TB in the ED, TB is often unsuspected and isolation measures are often not used . ED suspicion of TB is associated with more rapid isolation and treatment. South Med J, 1995 Sep, 88(9), 969 - 72 Fatal Bacillus cereus meningoencephalitis in an adult with acute myelogenous leukemia; Marley EF et al.; Bacillus cereus, a ubiquitous, endospore-forming, aerobic gram-positive bacillus, is primarily associated with toxin-mediated food poisoning . Frequently, isolates of Bacillus species from clinical specimens are discussed as contaminants . We report a rapidly fatal case of disseminated infection due to B cereus in a patient receiving induction chemotherapy for M0 acute leukemia . A short clinical syndrome of nausea and vomiting preceded neurologic symptoms . Autopsy showed extensive meningoencephalitis with subarachnoid hemorrhage and multiple liver abscesses . Areas of necrosis were devoid of any inflammatory response consistent with a severely immunocompromised state . The organism was isolated from immediate premortem and autopsy blood specimens . This case illustrates the possibility and severity of true B cereus infections in immunocompromised patients, the clinicopathologic features of which are as yet not well defined. J Infect Dis, 1995 Sep, 172(3), 698 - 705 Recombinant bacille Calmette-Guérin priming against measles; Fennelly GJ et al.; Live attenuated measles virus (MV) vaccines are ineffective in young infants because of neutralization by maternal antibody . An immunization strategy that may permit priming for T cell memory for MV in infants at or shortly after birth uses recombinant bacille Calmette-Guerin expressing the full-length MV nucleocapsid (N) protein (rBCG::N) . C3H/He mice immunized with rBCG::N developed T cell responses and ELISA antibodies to the N protein and low levels of neutralizing antibody after intracranial infection with MV strain CAM/R40 . There was considerable reduction in the virus titer recovered from brain homogenates, a decrease in the incidence and severity of histologic encephalitis, and a decrease in mortality in rBCG::N-printed C3H/He mice compared with control mice . Given the limitations of existing live attenuated MV vaccines, these results encourage the further testing of rBCG::N vaccines in primate models. J Biol Chem, 1995 Sep 1, 270(35), 20309 - 15 The CryIA(c) receptor purified from Manduca sexta displays multiple specificities; Masson L et al.; The kinetic binding characteristics of four Bacillus thuringiensis CryI insecticidal crystal proteins to a Cry-binding protein, purified from Manduca sexta brush-border vesicles, were analyzed by an optical biosensor . This 120-kilodalton binding protein, previously determined to be aminopeptidase N, was converted to a 115-kilodalton water-soluble form by removing the attached glycosylphosphatidylinositol anchor with phospholipase C . The solubilized form recognized the three major subclasses of CryIA toxins but not CryIC even though all four CryI proteins are toxic to larvae of M . sexta . CryIA(a) and CryIA(b) toxins bound to a single site on the solubilized aminopeptidase N molecule whereas CryIA(c) bound to two distinct sites . Apparent kinetic rate constants were determined for each binding reaction . All three CryIA toxins exhibited moderately fast on rates (approximately 10(-5) M-1 s-1) and a slow reversible off rate (approximately 10(-3) s-1) . Although the second CryIA(c)-binding site retained a moderately fast association rate, it was characterized by a rate of dissociation from the amino-peptidase an order of magnitude faster than observed for the other CryIA-binding sites . CryIA(c) binding to both sites was strongly inhibited in the presence of N-acetylgalactosamine (IC50 = 5 mM) but not N-acetylglucosamine, mannose, or glucose . CryIA(a) and CryIA(b) binding were unaffected in the presence of the same sugars . Our results serve to illustrate both the complexity and the diverse nature of toxin interactions with Cry-binding proteins. Int J Lepr Other Mycobact Dis, 1995 Sep, 63(3), 391 - 4 Relapses during long-term follow up with drug-susceptible M . leprae among multibacillary leprosy patients treated with multidrug therapy regimens; case reports; Thomas A et al.; A controlled clinical trial in highly bacilliferous multibacillary leprosy patients was initiated in 1977 . We report here two cases of relapse during long-term follow up of patients 15 years after the start of treatment . The patients reported here were treated with rifampin, isoniazid, clofazimine and dapsone for the first 3 months followed by clofazimine and dapsone until 84 months in one case; the other case received the same treatment but had received dapsone alone for 60-80 months . The relapses occurred 6(1/2) 7(1/2) years after therapy was discontinued. J Antibiot (Tokyo), 1995 Sep, 48(9), 967 - 72 Sattabacins and sattazolins: new biologically active compounds with antiviral properties extracted from a Bacillus sp; Lampis G et al.; A series of new compounds with antiviral properties were isolated from a Bacillus sp . strain B-60 . They were named sattabacin (1), hydroxysattabacin (2), sattazolin (3) and methylsattazolin (4) . The biologically active compounds were recovered from fermentation broth by ethyl acetate extraction and silica-gel column fractionation . Their antiviral activity was mainly expressed against the Herpes simplex viruses type 1 and 2 . The compound 3 showed a selective inhibition of protein synthesis in Herpesvirus-infected cells. Immunology, 1995 Sep, 86(1), 97 - 103 Cytokine profile of circulating T cells of leprosy patients reflects both indiscriminate and polarized T-helper subsets: T-helper phenotype is stable and uninfluenced by related antigens of Mycobacterium leprae; Misra N et al.; Cytokine profiles of circulating mononuclear cells were studied with the aim of delineating T-cell subsets in leprosy patients with active disease . Using reverse transcriptase-polymerase chain reaction (RT-PCR) for cytokine mRNA and enzyme-linked immunoassay (ELISA) for the secreted products, interferon-gamma (IFN-gamma), interleukin-4 (IL-4), IL-6 and granulocyte-macrophage colony-stimulating factor (GM-CSF) were studied . Three antigens, native Mycobacterium leprae, a recombinant antigen LSR/A15 of M . leprae and peptide 624 spanning 58-77 amino acids of the latter, were used to induce cytokine expression and release . Half of the subjects, irrespective of the clinical type or antigen used, showed a mixed T-helper type 0 (Th0)-like cytokine pattern, with evidence of the concomitant presence of IFN-gamma and IL-4 . The remainder showed a polarized pattern based on the type of leprosy . Lepromatous patients with disseminated disease had Th2-type cytokines, with IL-4 but not IFN-gamma . In contrast, tuberculoid leprosy patients with localized disease showed a Th1-like profile, with the presence of IFN-gamma but not IL-4 . Of interest was the stability of the Th phenotype for M . leprae-related antigens . Both the recombinant and the peptide antigens induced the same phenotype as the natural M . leprae bacillus in all except four of 45 leprosy patients. Microbiologia, 1995 Sep, 11(3), 343 - 50 Detection of Bacillus larvae spores in Argentinian honeys by using a semi-selective medium; Alippi AM; A semi-selective medium for the detection in Argentinian honeys of spores of Bacillus larvae, a pathogen of American foulbrood, was developed . The technique involves dilution of samples (1:2) in phosphate buffer, concentration of spores by centrifugation and heat treatment prior to inoculation . Two media (JNxPa and JNxPb) were prepared from J-agar, to which nalidixic acid and pipemidic acid were added . Both JNxP media were reliable for the isolation of B . larvae colonies and, at the same time, prevented the development of other Bacillus species which normally develop on the plates before B . larvae spores can germinate. PDA J Pharm Sci Technol, 1995 Sep-Oct, 49(5), 235 - 8 Heat resistance of Bacillus coagulans spores suspended in various parenteral solutions; Moldenhauer JE et al.; Bacillus coagulans, ATCC 51232 (formerly referred to as FRR B666), was previously reported to be a potentially excellent biological indicator . The organism was grown on a relatively simple sporulation media and the spores were evaluated for moist heat resistance in various parenteral solutions . Resistance was determined using the survivor curve method . For those solutions which appeared to be inhibitory to the outgrowth of Bacillus coagulans, ATCC 51232, further investigations were performed to evaluate the inhibitory effect . The results obtained demonstrated that the organism had the highest resistance in Calcium Gluconate Injection; USP (100 mg/mL) at 120 degrees C. Ann Clin Lab Sci, 1995 Sep-Oct, 25(5), 409 - 13 Granulomatous hepatitis as a complication of intravesical Bacillus Calmette-Guerin therapy for bladder carcinoma; Arzt MR et al.; Bacillus Calmette-Guerin (BCG) therapy has been proven to be effective treatment for superficial bladder carcinoma with complications that are usually minor and self limited . Although rare, granulomatous hepatitis can develop, and it is important to recognize it and treat it promptly . The hepatitis which occurs appears to be the result of a hypersensitivity reaction to antigens present in the BCG vaccine . Pathological findings include hepatocellular necrosis and pleomorphism, microgranulomas and non-caseous epitheliod granulomas with giant cells . A case of BCG-hepatitis, which developed in a 74-year-old white male following immunotherapy for superficial bladder carcinoma, is described. Nippon Hinyokika Gakkai Zasshi, 1995 Sep, 86(9), 1493 - 6 {Tuberculous spondylitis after intravesical BCG instillation: a case report}; Sugita Y et al.; We report a case of tuberculous spondylitis after intravesical bacillus Calmette-Guerin (BCG) instillation . A 71-year-old man was administered BCG (80 mg per week) for 8 weeks for prophylactic treatment of bladder cancer . After the first instillation he experienced miction pain, pollakisuria, and febrile episodes . Two months after the completion of BCG instillation, he complained to back pain and spinal X-ray showed a lytic lesion of Th7 vertebra . Diagnosis of metastatic transitional cell carcinoma was made based on MRI and bone scan . But pathological findings at laminectomy revealed tuberculous spondylitis . Antituberculous therapy (SM, RFP, and INH) was instituted and anterior supine fusion was performed . Now he is free from bladder cancer and tuberculous infection . Intravesical BCG instillation is effective for superficial bladder cancer, but it should be kept in mind that complications related to this treatment could occur and the adequate antituberculosis treatment has to be insisted if indicated. Gene, 1995 Aug 30, 162(1), 75 - 9 Nucleotide sequence analysis of the 23S ribosomal RNA-encoding gene of Bartonella bacilliformis; Minnick MF et al.; We report the cloning and characterization of the 23S ribosomal RNA (rRNA)-encoding gene (rDNA) of Bartonella bacilliformis (Bb) . The 2821-bp gene is preceded by an 11-nucleotide (nt) inverted repeat (IR) located 81 nt upstream in the tRNA(Ala)-23S rDNA intergenic spacer . The gene is followed by an 8-nt IR, five nt downstream in the 23S-5S rDNA intergenic spacer . The nt sequence of the Bb 23S rDNA is most similar to the 23S rDNA of Rhodopseudomonas palustris (Rp), with 85.4% sequence identity . The Bb 23S rDNA has 77.8% identity to the same gene from Escherichia coli (Ec) . Secondary structure predictions indicate that the large subunit (LSU) Bb rRNA contains two smaller stem-loops at nt 1459-1544 and 1658-1685, as compared to the corresponding loops from Ec (nt 1405-1597 and 1707-1751, respectively) . In addition, the Bb 23S rRNA has a large 72-nt stem-loop at nt 130-201, as compared to the Ec 18-nt stem-loop (nt 131-148) . There is no Bb homologue of the 15-nt stem-loop at the 3' end of the Ec molecule (nt 2791-2805) . The Bb 23S rRNA lacks the large stem-loop present in the LSU rRNA of closely related Rhodobacter sphaeroides (Rs) (nt position 1225-1331) which is thought to be involved in cleavage of 23S RNA precursor molecules into 16S and 14S rRNA species . This is the first LSU rDNA nt sequence for any member of the Bartonellaceae family. J Mol Biol, 1995 Aug 18, 251(3), 432 - 47 Unfolding simulations of the 85-102 beta-hairpin of barnase; Pugliese L et al.; Molecular dynamics simulations are used to investigate the unfolding reaction of an isolated beta-hairpin formed by residues 85 to 102 of barnase, a ribonuclease from Bacillus amyloliquefaciens . This peptide was considered following evidence from experimental studies that it may act as an initiation site for barnase folding by adopting a native-like conformation early during the folding process . Three successive molecular dynamics simulations of about 300 ps each were carried out for an all-atom model of the hairpin in water at 300 K, 450 K, and 600 K, respectively . A detailed analysis of all three simulations is presented . In particular we investigate the behavior of the backbone hydrogen bonds, and of hydrophobic interactions between side-chains, where distinction is made between contributions from native and non-native contacts, respectively . Furthermore, we investigate peptide water interactions and monitor the presence and size of empty cavities . The behavior of the hairpin in the three simulations, when considered sequentially, describes a process whereby a native-like conformation evolves to an unfolded state . Unfolding starts at the beginning of the 450 K simulation with the loss of two hydrogen bonds at the free hairpin extremities . At about the same time, the centrally located H-bonds are weakened and exchange more frequently with water, but the turn tightens up as the beta-sheet extends into the turn region . All this is accompanied by a volume expansion and the formation of a large hydrophobic side-chain cluster promoted by both native and highly fluctuating non-native apolar contacts involving residues 87 to 90 and 95 to 99 . This collapsed but more loosely packed state, essentially stabilized by hydrophobic interactions, is stable throughout the entire 450 K simulation and for about 150 ps at 600 K, after which point it proceeds rapidly to completely denatured conformations . This behavior presents clear analogies with known features of the unfolding reaction of complete proteins . It may indicate that this beta-hairpin has a well-defined conformation on its own, which would be in agreement with its role as an initiation site for folding. EMBO J, 1995 Aug 15, 14(16), 4056 - 64 X-ray crystallography shows that translational initiation factor IF3 consists of two compact alpha/beta domains linked by an alpha-helix; Biou V et al.; The structures of the two domains of translational initiation factor IF3 from Bacillus stearothermophilus have been solved by X-ray crystallography using single wavelength anomalous scattering and multiwavelength anomalous diffraction . Each of the two domains has an alpha/beta topology, with an exposed beta-sheet that is reminiscent of several ribosomal and other RNA binding proteins . An alpha-helix that protrudes out from the body of the N-terminal domain towards the C-terminal domain suggests that IF3 consists of two RNA binding domains connected by an alpha-helix and that it may bridge two regions of the ribosome . This represents the first high resolution structural information on a translational initiation factor. Biochem J, 1995 Aug 15, 310 ( Pt 1), 255 - 61 Structural analysis of inositol phospholipids from Trypanosoma cruzi epimastigote forms; Bertello LE et al.; Inositol phospholipids (IPL) from epimastigote forms of Trypanosoma cruzi have been investigated by metabolic labelling with {3H}palmitic acid and by GLC-MS analysis of the lipids obtained from non-labelled parasites . The IPL fraction was separated into phosphatidylinositol (PI) and inositol-phosphoceramide subfractions, the latter accounting for 80-85% of the total IPL . The neutral lipids released from the IPLs by PI-specific phospholipase C (PI-PLC) from Bacillus thuringiensis were analysed by silica-gel and reverse-phase TLC for the radioactive lipids and by GLC-MS for the non-radioactive samples . Ceramides containing dihydrosphingosine and sphingosine with C16:0 and C18:0 fatty acids were identified . The main component in the {3H}palmitic acid-labelled ceramides was palmitoyldihydrospingosine, while in the non-labelled sample the ceramides contained mainly sphingosine . This could reflect partial uptake of phospholipid from the medium . The PI contain both alkylacyl- and diacyl-glycerol lipids, with the ether lipid being more abundant . The latter was identified as 1-O-hexadecylglycerol esterified by C18:2 and C18:1 fatty acids . Interestingly, the same lipid had been identified in the anchor of the 1G7 glycoprotein of T . cruzi metacyclic forms. J Immunol, 1995 Aug 15, 155(4), 2057 - 66 Dissection of strain difference in acquired protective immunity against Mycobacterium bovis Calmette-Guérin bacillus (BCG) . Macrophages regulate the susceptibility through cytokine network and the induction of nitric oxide synthase; Yoshida A et al.; Protection against infection with intracellular pathogens operates in two stages, early innate resistance and late acquired protective immunity (API), in inbred mouse strains . Although both C57BL/10 (B10) and BALB/c mice bear the susceptible phenotype of innate resistance, Calmette-Guerin bacillus (BCG) vaccination generated efficient API in B10 but not in BALB/c mice . Employing a specific nitric oxide (NO) synthase inhibitor, we revealed that NO production plays a pivotal role in the API of B10 mice . Consistent with this, expressions of the inducible isoform of NO synthase (iNOS) protein and mRNA were significantly higher in the spleen of B10 mice than in that of BALB/c mice . Furthermore, IFN-gamma, a potent inducer of iNOS, and mRNAs for IL-12 (p40); an inducer of IFN-gamma and IL-2 were also vigorously expressed in the spleen of B10 mice compared with that of BALB/c mice . In an attempt to clarify the mechanism by which the different capacities for API are generated, we analyzed the cytokine network between T cells and macrophages in both B10 and BALB/c mice . We found that multiple functions of macrophages, which include capacities to express IL-12 (p40) mRNA in response to BCG and to express mRNAs for iNOS and IL-12 (p40) in response to IFN-gamma, were impaired in BALB/c mice as compared with B10 mice . However, T cells appeared to express comparable level of IFN-gamma mRNA in both strains when stimulated with IL-12 . Taken together, these results indicate that the macrophage functions play a pivotal role in both the induction and effector phases of API to determine the susceptibility of mice to BCG infection. FEBS Lett, 1995 Aug 14, 370(1-2), 93 - 6 Alteration in relative activities of phenylalanine dehydrogenase towards different substrates by site-directed mutagenesis; Seah SY et al.; Glycine-124 and leucine-307 of phenylalanine dehydrogenase from Bacillus sphaericus were altered by site-specific mutagenesis to the corresponding residues in leucine dehydrogenase: alanine and valine, respectively . These two residues have previously been implicated from molecular modelling as important in determining the substrate discrimination of the two enzymes . Single and double mutants displayed lower activities towards L-phenylalanine and enhanced activity towards almost all aliphatic amino acid substrates tested compared to the wild-type, thus confirming the predictions made from molecular modelling. Anal Biochem, 1995 Aug 10, 229(2), 313 - 6 Chromogenic assay for phospholipase C from Bacillus cereus; Hergenrother PJ et al.; A standard chromogenic assay for phosphatidylcholine determination has been modified in the development of a facile method for measuring the rate parameters (Km and Vmax) for phosphatidylcholine-preferring phospholipase C from Bacillus cereus (PLCBc) . The assay is applied to the determination of the Ki for a known inhibitor of PLCBc. Health Policy Plan, 1995 Sep, 10(3), 284 - 95 Increasing the utilization of cost-effective health services through changes in demand; Barlow R et al.; Attaining efficiency in a health care system with a budget constraint involves increasing the utilization of the most cost-effective services . This can be achieved by adjustments to prices, cost curves, or demand curves . In this paper, the potential for demand curve adjustments is examined by selecting two apparently cost-effective services (prenatal care and childhood immunization against tuberculosis), and analyzing the factors explaining their utilization . Data from recent household surveys in Burkina Faso and Niger are used . A multivariate analysis of utilization employs income, price, and taste variables . Utilization is highly sensitive to the distance which must be travelled to the health facility, a price, and taste variables . Utilization is highly sensitive to the distance which must be travelled to the health facility, a price variable . Members of certain ethnic groups tend to use the services less, other things being equal . The importance of demand-side factors like ethnicity points to certain kinds of policy interventions like information, education and communication activities which could increase the utilization of cost-effective servicesPIP: Two surveys undertaken by the Health Financing and Sustainability Project in Burkina Faso and Niger in 1994 provided the data for analysis . Prenatal care information was collected in households from women who had been pregnant during the previous 12 months . Bacillus Calmette-Guerin (BCG) immunization information was available only from the Niger survey . In Burkina Faso only 48% of women received prenatal care and only 33% did in the Niger sample . A multivariate analysis of utilization of these 2 services employed income, price, and taste variables (women's age, education, and ethnicity) . The low utilization of services was examined using independent variables . Regarding prenatal care the effect of income was visible in Niger, but it was absent in Burkina Faso . Ethnicity had an important effect on prenatal care . In Burkina Faso it was much less likely to get prenatal care for Fulani or Gourmantche women rather than Mossi . Also in Niger among the Fulani and Hausa low utilization was prevalent . The price variables also affected utilization, particularly the distance to the health facility . On the other hand, user charges stimulated utilization because enhanced services . Teenagers were less likely to get their babies immunized, although they were just as likely to get prenatal care . In Niger 26% of girls received BCG compared to 22% of boys . Part of the powerful effect of ethnicity was explained by French colonial rule, when Mossi and Zarma groups were incorporated into the colonial administration . This suggests the need for certain policy interventions, such as information, education, and communication activities to increase utilization of cost effective services . J Biol Chem, 1995 Aug 4, 270(31), 18615 - 25 The molecular cloning and characterization of BM1P1 and BM1P2 proteins, putative positive transcription factors involved in barbiturate-mediated induction of the genes encoding cytochrome P450BM-1 of Bacillus megaterium; He JS et al.; Analysis of a 1.3-kilobase segment of 5'-flanking DNA from the barbiturate-inducible P450BM-1 gene (CYP106) of Bacillus megaterium revealed two open reading frames . One, BM1P1, encodes 98 amino acids and is located 267 base pairs upstream from the sequence encoding cytochrome P450BM-1 but in the opposite orientation . The second, BM1P2 (88 amino acids), is 892 base pairs upstream from the P450BM-1 coding sequence and in the same coding strand . The expression of BM1P1 and BM1P2 was strongly stimulated in cells grown in the presence of pentobarbital, and the BM1P1 gene product exerted positive control on expression of P450BM-1 . When a 177-base pair fragment encompassing the overlapping promoter regions of the P450BM-1 and BM1P1 genes was used as a probe in DNA binding assays, the BM1P1 and BM1P2 gene products and Bm3R1 (the repressor protein regulating the barbiturate-mediated expression of P450BM-3) could bind individually, but the addition of BM1P1 or BM1P2 to a binding mixture containing Bm3R1 completely prevented the appearance of a Bm3R1 binding band . When a 208-base pair fragment containing a Barbie box sequence and located upstream of the 177-base pair fragment was used as a probe, only a Bm3R1 binding band was detected . Although neither BM1P1 and BM1P2 appeared to bind to this 208-base pair fragment, their presence strongly inhibited the binding of Bm3R1 to the same probe . The evidence suggests that BM1P1 and BM1P2 may, in part, act as positive regulatory proteins involved in the expression of the P450BM-1 gene by interfering with the binding of the repressor protein, Bm3R1, to the regulatory regions of P450BM-1. J Biol Chem, 1995 Aug 4, 270(31), 18606 - 14 Transcriptional regulation of the genes encoding cytochromes P450BM-1 and P450BM-3 in Bacillus megaterium by the binding of Bm3R1 repressor to Barbie box elements and operator sites; Liang Q et al.; We previously reported (Liang, Q., He, J.-S., and Fulco, A.J . (1995) J . Biol . Chem . 270, 4438-4450) that Bm3R1, a repressor regulating the expression of P450BM-3 in Bacillus megaterium, could bind to Barbie box sequences in the 5'-flanking regions of barbiturate-inducible genes . We've now shown that pentobarbital does not inhibit in vitro binding of Bm3R1 to the P450BM-3 and P450BM-1 Barbie boxes (BB3 and BB1), although the palindromic operator sequence (OIII) of P450BM-3 did have a strong competitive effect on such binding . G39E-Bm3R1, a mutant of Bm3R1, did not bind to either Barbie box . In the presence of Bm3R1, portions of the regulatory regions of P450BM-3 and P450BM-1 were protected from DNase I digestion . These included 11 of the 15 base pairs of BB3 plus 7 base pairs 3' to BB3, BB1 plus 16 base pairs 3' to BB1, and, in the 5'-flanking region of P450BM-1, segments covering most of two palindromic sequences (OII and OIII) of 24 and 52 base pairs . These DNase I-protected regions (including OIII) showed considerable sequence identity, especially in a conserved poly(A) motif . Barbiturates did not inhibit binding of Bm3R1 to OI.OII in vitro while G39E-Bm3R1 did not bind . The regulatory effects of Bm3R1 on P450BM-1 and P450BM-3 were also evaluated in vivo using heterologous chloramphenicol acetyltransferase constructs and Western blotting . In the G39E mutant strain, both P450BM-1 and P450BM-3 were constitutively expressed, and the regulatory proteins Bm1P1 and Bm3P1, although still pentobarbital-inducible, had significantly higher basal levels of synthesis . In toto, our results show that Bm3R1 represses both P450BM-1 and P450BM-3 expression and that it may effect this by coordinate binding to operator and Barbie box sequences to produce looping of the P450BM-1 and P450BM-3 regulatory regions through protein-protein interaction. J Biol Chem, 1995 Aug 4, 270(31), 18396 - 400 Molecular cloning and nucleotide sequences of the genes for two essential proteins constituting a novel enzyme system for heptaprenyl diphosphate synthesis; Koike-Takeshita A et al.; The genes encoding two dissociable components essential for Bacillus stearothermophilus heptaprenyl diphosphate synthase (all-trans-hexparenyl-diphosphate:isopentenyl-diphosphate hexaprenyl-trans-transferase, EC 2.5.1.30) were cloned, and their nucleotide sequences were determined . Sequence analyses revealed the presence of three open reading frames within 2,350 base pairs, designated as ORF-1, ORF-2, and ORF-3 in order of nucleotide sequence, which encode proteins of 220, 234, and 323 amino acids, respectively . Deletion experiments have shown that expression of the enzymatic activity requires the presence of ORF-1 and ORF-3, but ORF-2 is not essential . As a result, this enzyme was proved genetically to consist of two different protein compounds with molecular masses of 25 kDa (Component I) and 36 kDa (Component II), encoded by two of the three tandem genes . The protein encoded by ORF-1 has no similarity to any protein so far registered . However, the protein encoded by ORF-3 shows a 32% similarity to the farnesyl diphosphate synthase of the same bacterium and has seven highly conserved regions that have been shown typical in prenyltransferases (Koyama, T., Obata, S., Osabe, M., Takeshita, A., Yokoyama, K., Uchida, M., Nishino, T., and Ogura, K . (1993) J . Biochem . (Tokyo) 113, 355-363). Neurosci Lett, 1995 Aug 4, 195(2), 101 - 4 Monomers and dimers of acetylcholinesterase in human meningioma are anchored to the membrane by glycosylphosphatidylinositol; Saez-Valero J et al.; Amphiphilic monomers and dimers of acetylcholinesterase (AChE) and hydrophilic tetramers of butyrylcholinesterase (BuChE) were released by extracting human meningioma with Tris-saline and Tris-saline-Triton X-100 buffers . The amphiphilic or hydrophilic behavior of the AChE and BuChE forms was assessed by sedimentation analysis, hydrophobic chromatography and Triton X-114 phase-partitioning . A significant fraction of the amphiphilic AChE species was converted into hydrophilic components by incubation of the soluble enzyme with phosphatidylinositol-specific phospholipase C (PIPLC) from Bacillus thuringiensis, this fraction being increased by a double treatment with PIPLC and alkaline hydroxylamine . A significant amount of the membrane-bound AChE was released by incubation with PIPLC . These results demonstrate that AChE forms in meningioma are attached to the membrane via glycosylphosphatidylinositol, although part of the enzyme forms are resistant to PIPLC. Singapore Med J . 1995 Aug;36(4):446. Melioidosis presenting as epididymo-orchitis; Koh KB; We report a rare case of suppurative epididymo-orchitis caused by Pseudomonas pseudomallei in a 56-year-old male . This is a gram negative bacillus found mainly in tropical zones . Diagnosis was reached by culture of the organisms after drainage of the scrotal abscess, and the patient was treated by a course of oral chloramphenicol 500 mg qid for 6 months. J Chemother, 1995 Aug, 7(4), 338 - 43 Once-daily dosing regimen for aminoglycoside plus betalactam combination therapy of serious lower respiratory tract infections; Paradisi F et al.; Aminoglycosides are important antibacterial agents for treatment of serious gram-negative bacillary infections including lower respiratory tract infection . Once-daily aminoglycosides result in higher peak and lower trough plasma concentrations than conventional multiple daily dosing regimens; once-daily aminoglycoside therapy is equally effective, generally less toxic and much less expensive and therefore this regimen is more and more frequently used for treatment of suspected or confirmed gram-negative bacillary infections and of febrile episodes in neutropenic patients, in particular in combination with an appropriate betalactam antibiotic . Despite the lack of studies on this topic, once-daily aminoglycosides in combination with a betalactam agent can be used in subjects with lower respiratory tract infection, including patients with cystic fibrosis, in which tobramycin appears to be the aminoglycoside antibiotic of choice. Clin Infect Dis, 1995 Aug, 21(2), 352 - 60 Neuroretinitis, aseptic meningitis, and lymphadenitis associated with Bartonella (Rochalimaea) henselae infection in immunocompetent patients and patients infected with human immunodeficiency virus type 1; Wong MT et al.; Bartonella (Rochalimaea) henselae causes a variety of diseases, including bacillary angiomatosis, peliosis hepatis, lymphadenitis, aseptic meningitis with bacteremia, and cat-scratch disease (CSD) . Cases of B . henselae-related disease were collected from September 1991 through November 1993 . Patients with suspected CSD, unexplained fever and lymphadenitis, or suspected B . henselae infection who were seen in the Infectious Diseases Clinic at Wilford Hall Medical Center (Lackland Air Force Base, TX) underwent physical and laboratory examinations . In addition to three previously described cases, 23 patients with R . henselae-related infection were identified . The patients included 19 immunocompetent individuals presenting with lymphadenitis (11), stellate neuroretinitis (5), Parinaud's oculoglandular syndrome with retinitis (1), chronic fatigue syndrome-like disease (1), and microbiologically proven adenitis without the presence of immunofluorescent antibodies to B . henselae (1) and four patients infected with human immunodeficiency virus type 1 presenting with isolated lymphadenitis (1), diffuse upper-extremity adenitis (1), neuroretinitis (1), and aseptic meningitis (1) . A couple with neuroretinitis and their pet cat, a persistently fatigued patient, and a patient with Parinaud's oculoglandular syndrome were shown to have bacteremia . Tissue cultures were positive for B . henselae in three recent cases of adenitis . Twenty-two patients were exposed to cats . This series further demonstrates the similarities between B . henselae-related diseases and CSD and identifies several new syndromes due to B . henselae. Braz J Med Biol Res, 1995 Aug, 28(8), 843 - 51 New vaccines against tuberculosis; Silva CL; It has proved difficult to vaccinate effectively against tuberculosis with mycobacterial components or even with whole dead mycobacteria; protection was always inferior to that obtained with the live attenuated vaccine known as bacillus Calmette-Guerin (BCG) . We have found that this may no longer be the case . Expression of the gene for a single mycobacterial antigen (Mycobacterium