Bacillus

Appl Environ Microbiol, 1995 Apr, 61(4), 1649 - 52
Cocultivation of the amoeba Naegleria fowleri and the amoebicin- producing strain Bacillus licheniformis M-4; Lebbadi M et al.; Antagonism between Bacillus licheniformis M-4 and the pathogenic amoeba Naegleria fowleri HB-1 during cocultivation was influenced by the composition of the medium and the initial amoeba/bacterium ratio . While a ratio of 50 caused complete lysis of amoebae in soil extract with 0.3% glucose (SEG) before 72 h, this ratio had to be at least 12-fold lower in order to obtain similar results in Cline medium . Sporulation of B . licheniformis M-4 took place much earlier in SEG . Amoebicin production was stimulated by the presence of amoebae by either shortening the time of production (as in SEG) or increasing the amount of amoebicins released (as in Cline medium) . Electron microscopy showed that amoebae cocultivated in the Cline medium contained bacteria enclosed in digestive vacuoles, while amoebae from SEG cocultures did not.

Appl Environ Microbiol, 1995 Apr, 61(4), 1469 - 74
Nucleotide sequence and expression of kerA, the gene encoding a keratinolytic protease of Bacillus licheniformis PWD-1; Lin X et al.; Bacillus licheniformis PWD-1 (ATCC 53757) secretes keratinase, a proteolytic enzyme which is active on whole feathers . By amino acid sequence similarity and phenylmethylsulfonyl fluoride inhibition, the keratinase was demonstrated to be a serine protease . The entire nucleotide sequence of the coding and flanking regions of the keratinase structure gene, kerA, was determined . A fixed oligonucleotide primer derived from the N-terminal sequence of the purified enzyme and a second random oligonucleotide primer were used in a procedure called PCR walking, which was developed to amplify and sequence the upstream and downstream regions of kerA . Another method, PCR screening, was conducted with a lambda phage vector with inserted PWD-1 genomic DNA fragments as templates and with the known sequences of the vector arms and the N-terminal sequence of the enzyme as primers . PCR amplification and sequence analysis of the lambda library completed the entire kerA sequence and established a set of gene deletions . The kerA gene shares a 97% sequence identity with the gene encoding subtilisin Carlsberg from B . licheniformis NCIMB 6816 . The putative promoters, ribosome binding sites, and transcriptional terminators are also similar in these two bacteria . The deduced amino acid sequences indicate only three amino acid differences between the two mature proteases . Northern (RNA) analysis demonstrates that transcriptional regulation controls kerA expression on different growth media.

Am J Trop Med Hyg, 1995 Apr, 52(4), 354 - 9
A review of bartonellosis in Ecuador and Colombia; Alexander B; A review of the literature regarding bartonellosis or Carrion's disease in Colombia and Ecuador is presented, together with observations made by the author in areas of both countries from which the disease has been recorded . There is evidence from pre-Columbian artifacts that verruga peruana, the cutaneous form of the disease, was present in Ecuador at least 1,000 years prior to the arrival of Europeans . These artifacts were discovered in the coastal province of Manabi, a low-lying area very different from the high Andean valleys of Peru with which bartonellosis is normally associated . Most of the cases recorded in recent years from this coastal area . The disease does not appear to have occurred in Colombia before the 1930s and only one case has been reported during the past 40 years . The possibility of many more subclinical cases being present in both Ecuador and Colombia is discussed, together with the possibility that the acquired immunodeficiency syndrome epidemic will reveal a higher prevalence among the inhabitants of endemic areas than previously suspected . Although the suspected vector of Bartonella bacilliformis, the sand fly Lutzomyia verrucarum, has not been recorded from Ecuador or Colombia, related species are present in endemic areas and may be involved in transmission.

Int J Syst Bacteriol, 1995 Apr, 45(2), 409 - 11
Antibiotic susceptibility as a taxonomic characteristic of the genus Bacillus; Reva ON et al.; A large number of Bacillus strains assigned to different species were tested to determine their susceptibilities to antibiotics . Some clear differences between species were observed . The antibiotic susceptibilities of strains isolated from natural sources seemed to be stable and to reflect adaptation of the strains to specific conditions in certain ecological niches . A method for data processing which can be used for rapid species identification is described.

Int J Syst Bacteriol, 1995 Apr, 45(2), 212 - 7
Random amplified polymorphic DNA fingerprinting of mosquito-pathogenic and nonpathogenic strains of Bacillus sphaericus; Woodburn MA et al.; Random amplified polymorphic DNA fingerprinting was used to examine 31 mosquito-pathogenic and 14 nonpathogenic strains of Bacillus sphaericus . We verified that DNA bands that migrated the same distance in an agarose gel were homologous by using PCR-generated probes made from the random amplified polymorphic DNA bands . The band patterns obtained with eight primers were analyzed by using the Jaccard coefficient and unweighted pair group with arithmetic average clustering . Pathogenic strains belonging to DNA homology group IIA were similar to strains belonging to nonpathogenic homology groups at an average level of similarity of 6.3% . Individual serotypes were clearly identified among the pathogenic strains . This suggests that there is overall genetic homogeneity among strains within serotypes . It is also consistent with the uniform toxicity pattern found for each serotype (unlike the toxin diversity found in Bacillus thuringiensis serotypes) . These results, together with DNA homology data, support the proposal that a new species should be described for the pathogenic strains.

J Econ Entomol, 1995 Apr, 88(2), 270 - 7
Increased efficacy of Bacillus thuringiensis subsp . kurstaki in combination with tannic acid; Gibson DM et al.; We identified tannic acid as an inexpensive additive that increased the efficacy of sublethal concentrations of Bacillus thuringiensis subsp . kurstaki (Berliner) . Tannic acid mimicked the active constituents contained in an aqueous, tannin-rich extract of Taxus baccata (L.) bark that retarded development of Heliothis virescens (F.) larvae at 10,000 ppm; most larvae remained in first and second stage when treated with 250-10,000 ppm of tannic acid . Instar development of Trichoplusia ni (Hubner) larvae was affected in a concentration-dependent manner by 2.5-500 ppm of tannic acid . In subsequent bioassays, tannic acid at 25-500 ppm in combination with B . thuringiensis (1.63 micrograms {AI}/ml diet) yielded mean mortalities of 57-75%, whereas treatments with B . thuringiensis alone produced 10% mortality . Mean mortalities in the 3.0, 4.5, and 6.75 micrograms (AI) B . thuringiensis per milliliter of diet treatments (5.5; 8.0, and 30%, respectively) were significantly higher in the presence of 250 and 2,500 ppm tannic acid; in these treatments we observed 78-94% mortality . Addition of tannic acid increased the activity of concentrations of 3-4.5 micrograms (AI) B . thuringiensis per milliliter of diet to approximately that of a concentration of 13 micrograms (AI) B . thuringiensis per milliliter of diet alone (85-95% mortality) . Although deaths caused by a formulation of B . thuringiensis + tannic acid occurred more slowly than with high rates of B . thuringiensis alone, such formulations would have the advantages of arresting development, minimizing foliar damage, and decreasing the concentration of B . thuringiensis used.

J Bacteriol, 1995 Apr, 177(8), 1981 - 8
Analysis of a novel gene and beta-galactosidase isozyme from a psychrotrophic Arthrobacter isolate; Gutshall KR et al.; We have characterized a new psychrotrophic Arthrobacter isolate which produces beta-galactosidase isozymes . When DNA from this isolate was transformed into an Escherichia coli host, we obtained three different fragments, designated 12, 14, and 15, each encoding a different beta-galactosidase isozyme . The beta-galactosidase produced from fragment 12 was of special interest because the protein subunit was smaller (about 71 versus 116 kDa) than those typically encoded by the lacZ family . The isozyme encoded by fragment 12 was purified, and its activity and thermostability were examined . Although the enzyme is highly specific towards beta-D-galactoside substrates, its levels in the isolate do not increase in cells grown with lactose . Nucleotide sequence determination showed that the gene encoding isozyme 12 is not similar to the other members of the lacZ family but has regions similar to beta-galactosidase isozymes from Bacillus stearothermophilus and B . circulans . Addition of the isozyme 12 sequence to the database made it possible to examine these enzymes as possible members of a new, separate family . Our analysis of this new family showed some conserved amino acids corresponding to the lacZ acid-base catalytic region but no homology with the nucleophilic region . On the basis of these comparisons, we designated this a new lacG family.

J Clin Psychiatry, 1995 Apr, 56(4), 161 - 6
Bacillary angiomatosis: a treatable cause of acute psychiatric symptoms in human immunodeficiency virus infection; Baker J et al.; BACKGROUND: Bacillary angiomatosis is a systemic infection that has been most commonly reported in the setting of immunosuppression, especially human immunodeficiency virus (HIV) disease . METHOD: We report two patients who had bacillary angiomatosis who presented with psychiatric symptoms . RESULTS: The first patient presented with marked exacerbation of previous depressive disease . The second patient presented with new psychotic symptoms . In both cases psychiatric symptoms did not resolve until antibiotic treatment was given . CONCLUSION: Our report expands the clinical spectrum of bacillary angiomatosis and identifies a new cause of treatable psychiatric disease in HIV-infected persons.

Br J Cancer, 1995 Apr, 71(4), 801 - 7
Bacillus Calmette-Guerin (BCG) enhances monocyte- and lymphocyte-mediated bladder tumour cell killing; Pryor K et al.; A cytotoxicity assay was used to study the action of bacillus Calmette-Guerin (BCG) and cytokines on four human bladder cancer cell lines . Monocytes and lymphocytes from peripheral blood were incubated with or without BCG or cytokines for 24 h, after which {3H}thymidine-labelled target cells were added and the 72 h percentage specific release determined . BCG had a direct cytotoxic effect against tumour cells and significantly enhanced monocyte/macrophage and enhanced lymphocyte cytotoxicity against one cell line (UCRU-BL-17) . Supernatants (SNs) from BCG-activated monocytes/macrophages and lymphocytes increased the percentage specific release of {3H}thymidine from UCRU-BL-17 cells . Interferon alpha (IFN-alpha) and interleukin 2 (IL-2) were cytotoxic towards UCRU-BL-17 . No synergy occurred between BCG and cytokines at the concentrations tested . The results suggest that BCG is superior to IFN-alpha, interferon gamma (IFN-gamma) and IL-2 in enhancing cell-mediated cytotoxicity.

Chest, 1995 Apr, 107(4), 1032 - 4
Sterilization of talc for pleurodesis . Available techniques, efficacy, and cost analysis; Kennedy L et al.; Although talc has been used as a pleurodesis agent since 1935, a sterilization protocol has not been established . We obtained USP asbestos-free talc from six different suppliers and sterilized each using dry heat, gamma irradiation, and ethylene oxide gas . Aerobic, anaerobic, and fungal cultures were obtained prior to sterilization, and 1, 30, and 90 days after sterilization . Bacillus species were cultured from all six unsterilized specimens and coagulase-negative Staphylococcus grew from two unsterilized specimens . No growth of organisms was found following any method of sterilization . The cost of sterilization per 5-g packet of talc was $4.74, $7.85, and $16.25 for heat, ethylene oxide, and gamma irradiation, respectively . In conclusion, untreated talc is not sterile . Sterilization by prolonged dry heat exposure, ethylene oxide gas, and gamma irradiation are all effective, with dry heat being the least expensive.

Clin Exp Immunol, 1995 Apr, 100(1), 26 - 31
Induction of tumour necrosis factor-alpha (TNF-alpha) mRNA in bladders and spleens of mice after intravesical administration of bacillus Calmette-Guérin; Shin JS et al.; Intravesical bacillus Calmette-Guerin (BCG) therapy is highly effective in the therapy of carcinoma in situ of the bladder, but the mechanism of BCG immunotherapy is not clearly understood . We studied the production of TNF-alpha in spleens and bladders of mice after intravesical BCG or BCG/interferon-gamma (IFN-gamma) instillation . Significant change of TNF-alpha mRNA expression of spleens and bladders of C3H/He mice was observed after intravesical BCG instillation, although intravesical IFN-gamma therapy 3 days after BCG instillation to maintain the activated state of monocyte/macrophage lineage cells did not show a significant change of TNF-alpha mRNA, compared with that of BCG therapy alone . Maximal production of TNF-alpha mRNA in spleens of mice was seen after the first or second intravesical BCG instillation, and production of TNF-alpha mRNA in bladders was also increased after intravesical BCG instillation . The increment of TNF-alpha production by BCG stimulation in HL-60, a promyelocytic leukaemic cell line, and peripheral blood mononuclear cells in vitro may support the in vivo effect of BCG therapy on the bladder . These data show that local production of TNF-alpha as well as systemic production by intravesical BCG treatment may correlate with one of the mechanisms of BCG immunotherapy of superficial bladder cancer.

Am J Infect Control, 1995 Apr, 23(2), 152 - 5
Implementing a tuberculosis control program; Williams J et al.; Between January 1989 and December 1990, 26 patients acquired multidrug-resistant tuberculosis at our institution . Their exposures occurred when they were admitted to a ward where a patient with acid fast bacillus smear-positive pulmonary tuberculosis was also admitted . In 20 cases, the infectious patients were not isolated until the sputum smears were positive . When the outbreak was recognized in the spring of 1990, the infection control department undertook a risk assessment and instituted measures that would become the tuberculosis control program . Since then, administrative and environmental controls have been implemented, education programs are ongoing, personal protective equipment is in use, and a more aggressive employee health testing program is underway . The steps we took and the barriers we had to overcome to implement our plan are included in this article.

Arukoru Kenkyuto Yakubutsu Ison, 1995 Apr, 30(2), 69 - 79
{Effect of Bacillus natto-fermented product (BIOZYME) on blood alcohol, aldehyde concentrations after whisky drinking in human volunteers, and acute toxicity of acetaldehyde in mice}; Sumi H et al.; Effects of Bacillus natto-fermented product (BIOZYME) on blood alcohol and aldehyde concentrations after drinking whisky (corresponding to 30-65 ml ethanol) were studied in 21 healthy volunteers . When 100 ml of BIOZYME was orally administrated to the volunteers before drinking whisky, the time delay of both blood factors to attain maximum concentrations were observed . The maximum decrease in blood alcohol and aldehyde concentrations were about 23% and 45% (p < 0.005), respectively, at 1 hr after drinking whisky . The aldehyde lowering effect of BIOZYME was continued for at least 4 hr after whisky drinking . Concentration of the breath alcohol was also sharply decreased by BIOZYME administration . The breath alcohol concentration in the administered group (0.18 +/- 0.11 mg/l) was found to be lowered about 44% than that of the control group (0.32 +/- 0.11 mg/l) (p < 0.0005, n = 21), at 1 hr after drinking whisky . In acute toxicity experiments of aldehyde in mice (12 mmol AcH/mg), BIOZYME showed the survival effect as with alpha-D-Ala (134% increase of the living, at 40 min after i.p . administration) (p < 0.005, n = 22) . These findings reveal the Bacillus natto produced BIOZYME as a reasonable, safety and useful anti-hangover agent.

Mol Microbiol, 1995 Apr, 16(2), 365 - 72
Saturation of penicillin-binding protein 1 by beta-lactam antibiotics in growing cells of Bacillus licheniformis; Lepage S et al.; With the help of a new highly sensitive method allowing the quantification of free penicillin-binding proteins (PBPs) and of an integrated mathematical model, the progressive saturation of PBP1 by various beta-lactam antibiotics in growing cells of Bacillus licheniformis was studied . Although the results confirmed PBP1 as a major lethal target for these compounds, they also underlined several weaknesses in our present understanding of this phenomenon . In growing cells, but not in resting cells, the penicillin target(s) appeared to be somewhat protected from the action of the inactivators . In vitro experiments indicated that amino acids, peptides and depsipeptides mimicking the peptide moiety of the nascent peptidoglycan significantly interfered with the acylation of PBP1 by the antibiotics . In addition, the level of PBP1 saturation at antibiotic concentrations corresponding to the minimum inhibitory concentrations was not constant, suggesting that additional, presently undiscovered, factors might be necessary to account for the experimental observations.

Int J Food Microbiol, 1995 Apr, 25(2), 131 - 9
Prevalence of Bacillus cereus in selected foods and detection of enterotoxin using TECRA-VIA and BCET-RPLA; Rusul G et al.; Enterotoxigenic Bacillus cereus was detected in cooked foods (17), rice noodles (3), wet wheat noodles (2), dry wheat noodles (10), spices (8), grains (4), legumes (11) and legume products (3) . One hundred ninety-four (42.3%), 70 (15.3%) and 23 (5.2%) of the 459 presumptive B . cereus colonies isolated from PEMBA agar were identified as B . cereus, Bacillus thuringiensis and B . mycoides, respectively . B . cereus isolates were examined for growth temperature, pH profile and enterotoxin production using both TECRA-VIA and BCET-RPLA kits . One hundred seventy-eight (91.8%) and 164 (84%) of the strains were enterotoxigenic as determined using TECRA-VIA and BCET-RPLA, respectively . Eighty-two (50%) of the enterotoxigenic strains were capable of growing at 5 degrees C, and 142 (86.6%) grew at 7 degrees C within 7 days of incubation . The enterotoxigenic strains did not grow at pH 4.0 but 69 (42.0%) of the strains were able to grow at pH 4.5 within 7 days at 37 degrees C . The isolates were resistant to ampicillin (98.8%), cloxallin (100%) and tetracycline (61.0%), and susceptible to chloroamphenicol (87%), erythromycin (77.4%), gentamycin (100%) and streptomycin (98.7%).

Indian J Biochem Biophys, 1995 Apr, 32(2), 100 - 5
Activity and stability of Bacillus cereus penicillinase entrapped in aerosol OT reverse micelles; Chakravarty K et al.; The kinetics of enzyme catalyzed hydrolysis of penicillin G and stability of the enzyme alpha-penicillinase, entrapped in aerosol OT reverse micellar droplets have been investigated spectrophotometrically . Various physical parameters, such as, water pool size (related to Wo), pH and temperature, were optimized for maximum activity of penicillinase in water/aerosol OT/isooctane reverse micelles . The enzyme showed maximum activity of Wo - 14 and pH, 7.0 . At any temperature the enzyme was to be more active in reverse micelles than in aqueous solution . At optimum conditions of Wo, pH and temperature the enzyme was 100% more active in reverse micelles than its maximum activity in aqueous solution . In both the systems, the activity starts falling at and above 25 degrees C . CD Spectral studies showed that the enzyme in reverse micelles possesses more helical structure than it has in aqueous solution and at the optimum conditions in which it showed maximum activity, the alpha-helicity was also maximum . The enzyme was very stable in reverse micelles at and above room temperature compared to the same in aqueous solution.

Biotechnol Appl Biochem, 1995 Apr, 21 ( Pt 2), 233 - 43
Cyclodextrin glycosyltransferase may be the only starch-degrading enzyme in Bacillus macerans; Nogrady N et al.; Cyclodextrin glycosyltransferase (CGTase) was released into the culture fluid by Bacillus macerans predominantly in the late stationary phase of growth and during autolysis in the presence of either glucose or starch as a carbon source . In both cases significant soluble intracellular enzyme activity could be observed in the early stationary phase, and a low non-soluble intracellular CGTase activity could be demonstrated also in the exponential growth phase in the presence of starch . At the end of the exponential phase the non-soluble specific intracellular enzyme activity was found to be constant with a value of 0.63 +/- 0.06 nkat/10(9) viable cells . Since amylase activity could not be detected in any intracellular or extracellular sample taken at any culture time, we conclude that cellbound CGTase is the only starch-digesting enzyme in growing B . macerans and, hence, may be fully responsible for the degradation of starch in the culture fluid.

Proc Natl Acad Sci U S A, 1995 Mar 28, 92(7), 2805 - 9
Serological response patterns of melanoma patients immunized with a GM2 ganglioside conjugate vaccine; Kitamura K et al.; Gangliosides, such as GM2, GD2, GD3, and 9-O-acetyl GD3, are receiving attention as targets for antibody-based and vaccine-based therapies of melanoma . GM2 appears to be a particularly immunogenic ganglioside in humans, as indicated by the presence of naturally occurring IgM anti-GM2 antibodies in approximately 5% of humans and the fact that immunization with irradiated GM2-expressing melanoma cells or purified GM2 adherent to bacillus Calmette-Guerin elicits GM2 antibodies of low to moderate titers in a high proportion of vaccinated patients . To develop vaccines that consistently induce high titers of IgM as well as IgG anti-GM2 antibodies, vaccines containing GM2 conjugated to keyhole limpet hemocyanin as the carrier protein and QS-21 as the adjuvant have been constructed . The serological response of vaccinated patients was monitored by ELISA using purified GM2 ganglioside for IgM and IgG anti-GM2 antibodies and for GM2 cell surface-reactive antibodies by immune adherence assays and cytotoxic tests (IgM antibodies) and mixed hemadsorption assays (IgG antibodies) . The majority of vaccinated patients developed IgM and IgG antibodies detectable by ELISA . In most cases, the results of IgM ELISA correlated with assays for cell surface-reactive IgM antibodies . This was not true for IgG anti-GM2 antibodies, where strong discrepancies were seen between high titers in ELISA and little or no reactivity in mixed hemadsorption tests for cell surface-reactive antibodies . These IgG antibodies (and the less frequent IgM antibodies that show similar discrepancies) may be directed against GM2 determinants that are buried, hidden, or not present on GM2-expressing target cells . With regard to a major objective of ganglioside vaccines--i.e., generation of cytotoxic antibodies--the GM2-keyhole limpet hemocyanin/QS-21 vaccine is clearly superior to the previously tested GM2/bacillus Calmette-Guerin vaccine . However, variability in patient response and lack of persistence of high-titered IgM cytotoxic antibodies in many patients are problems that remain to be solved.

Biochim Biophys Acta, 1995 Mar 22, 1234(2), 173 - 83
Purification of deformin, an extracellular protein synthesized by Bartonella bacilliformis which causes deformation of erythrocyte membranes; Xu YH et al.; A factor capable of deforming erythrocyte membranes, found in the culture supernatants of Bartonella bacilliformis, was purified 1840-fold using hydrophobic, ion exchange and gel exclusion chromatography . The final fractions contained a single detectable polypeptide species, referred to as deformin, having a molecular weight of 67000 by SDS-PAGE and a native molecular weight of 130,000 by gel exclusion chromatography or velocity sedimentation in a glycerol gradient . Erythrocytes treated with deformin acquire trenches, indentations, and invaginations which could be reversed by vanadate, dilauroylphosphatidylcholine (DLPC), or by raising the internal Ca2+ concentrations with the inophore A23187 . Internal vacuoles also form . Erythrocytes treated with trypsin or neuraminidase are much more sensitive to deformin than untreated erythrocytes; erythrocytes treated with phospholipase D are less sensitive to deformin . This protein may play a role in causing the severe anemia which can result as a consequence of infection by B . bacilliformis.

Med Clin (Barc), 1995 Mar 18, 104(10), 365 - 8
{The epidemiology of tuberculosis in El Ferrol}; Garcia Rodriguez JF et al.; BACKGROUND: This study was undertaken to know the frequency of tuberculosis in El Ferrol and to contribute to the knowledge of the situation in Spain . METHODS: A retrospective study of all the cases of tuberculosis diagnosed in the Hospital A . Marcide-Novoa Santos (El Ferrol, Spain) from 1990 to 1993 was performed . RESULTS: Seven hundred twenty-four patients were diagnosed, with a mean annual prevalence of 83.3/100,000 inhabitants . Six hundred sixty-four cases (430 males {64.8%}) were evaluated . The mean age was 35.5 +/- 19 years with 58.9% under the age of 35 . 98.7% of the patients lived in the health care area and 73.2% were admitted, with 13.7% having previous history of tuberculosis . Sixty-one cases (11.1%; Cl: 8.25-13.7) had HIV infection . Diagnosis was microbiological in 505 cases (76%), anatomopathological in 60 (9%) and in 99 (14.9%) diagnosis was achieved by clinical and radiological criteria . Pulmonary localization (67.2%) was the most frequent form and was predominant in males, while lymph node and osteoarticular localizations were more frequent in women . The incidence of bacilliferous patients was 30.7/100,000 inhabitants . A delay of more than one month took place in the diagnosis of 66.4% of the bacilliferous patients . CONCLUSIONS: The incidence of tuberculosis in El Ferrol is very high with an important delay in the diagnosis of bacilliferous patients . The high percentage of patients admitted to hospital carries considerable costs in the treatment of the disease.

J Biol Chem, 1995 Mar 17, 270(11), 6412 - 9
Mutations in domain I of Bacillus thuringiensis delta-endotoxin CryIAb reduce the irreversible binding of toxin to manduca sexta brush border membrane vesicles; Chen XJ et al.; Site-directed mutagenesis was used to generate CryIAb mutants at the selected N-terminal positions to study the function of domain I . Structurally stable mutant proteins were tested for toxicity, receptor binding kinetics, and pore function . Substitutions of tyrosine at position 153 with arginine (Y153R) or alanine (Y153A) did not affect toxicity appreciably, whereas replacing this tyrosine with aspartic acid (Y153D) resulted in a great loss of toxicity . Mutation of alanine at position 92 to glutamic acid (A92E) almost completely abolished toxicity . The initial receptor binding was unchanged as measured by competition binding assays among all mutant proteins . Reduced pore function, however, was observed for mutants A92E and Y153D as tested by voltage clamping . Further studies with specially designed association and dissociation binding assays showed that irreversible binding of these two mutant toxins to Manduca sexta brush border membrane vesicles was significantly reduced . The decrease in irreversible binding was correlated with the changes in toxicity and may reflect a severely disturbed membrane insertion process in these two mutant toxins, leading to reduced pore function and toxicity . The results support the model that domain I is involved in membrane integration and pore formation.

J Immunol, 1995 Mar 15, 154(6), 2753 - 63
Involvement of IFN-gamma in Bacillus Calmette-Guérin-induced but not in tumor-induced sensitization to TNF-induced lethality; Cauwels A et al.; In healthy mice, murine (m) TNF is fairly lethal, whereas human (h) TNF (a selective murine TNF-R55 agonist) is rather harmless . However, we and others observed that mice suffering from a bacterial infection, such as Bacillus Calmette-Guerin (BCG), or bearing i.m . some types of tumor, develop a hypersensitivity to the IL-6-inducing and lethal properties of hTNF . This is a cardinal problem as it severely limits the potential use of hTNF-R55-specific agonists for systemic treatment of human cancer . Using mice carrying a targeted disruption in the gene encoding the IFN-gamma receptor (IFN-gamma Ro/o), we here report that endogenous IFN-gamma plays a crucial role in the development of TNF hypersensitivity during BCG infection . Indeed, both the lethality and the IL-6 induced by hTNF were drastically reduced in IFN-gamma Ro/o mice as compared with control mice . These results demonstrate that the enhancement of TNF effects is at least an equally important mechanism by which IFN-gamma contributes to BCG-induced hypersensitivity as the previously described augmentation of TNF production . Experiments in athymic nude mice, either depleted of NK cells or not, revealed that the latter cell population is an important source of the sensitizing IFN-gamma during BCG infection . In contrast, IFN-gamma Ro/o mice were as susceptible as control mice to the sensitizing effects of tumors . mTNF, which interacts with both mTNF-R55 and mTNF-R75 and causes lethality on its own, is as toxic in IFN-gamma Ro/o mice as in wt control mice; this means that TNF-induced IFN-gamma does not play a role in mTNF-induced lethality.

Biochem J, 1995 Mar 15, 306 ( Pt 3), 727 - 33
Interaction of component enzymes with the peripheral subunit-binding domain of the pyruvate dehydrogenase multienzyme complex of Bacillus stearothermophilus: stoichiometry and specificity in self-assembly; Lessard IA et al.; The interaction between the pyruvate decarboxylase (E1) component and a di-domain (lipoyl domain plus peripheral subunit-binding domain) from the dihydrolipoyl acetyltransferase (E2) component of the Bacillus stearothermophilus pyruvate dehydrogenase multienzyme complex was investigated . Only 1 mol of di-domain (binding domain) was bound to 1 mol of heterotetrameric E1 (alpha 2 beta 2) and the binding was without effect on the kinetic activity of E1 . Similarly, the di-domain bound to separate E1 beta subunits at a maximal polypeptide chain ratio of 1:2, but no detectable interaction was found with the E1 alpha subunit . However, addition of the monomeric E1 alpha subunit to an E1 beta-di-domain complex generated a fully functional E1 (alpha 2 beta 2)-di-domain complex, indicating that the E1 beta subunit plays the critical part in binding the E1 component to the di-domain and suggesting that no chaperonin is needed in vitro to promote the assembly of the three separate proteins . Mixing the E1 and dihydrolipoyl dehydrogenase (E3) components in the presence of di-domain revealed that E1 and E3 cannot bind simultaneously to the same molecule of di-domain, a new feature of the assembly pathway and an important factor in determining the ultimate structure of the assembled enzyme complex.

Biochemistry, 1995 Mar 14, 34(10), 3368 - 76
Site-directed mutations in tyrosine 195 of cyclodextrin glycosyltransferase from Bacillus circulans strain 251 affect activity and product specificity; Penninga D et al.; Tyrosine 195 is located in the center of the active site cleft of cyclodextrin glycosyltransferase (EC 2.4.1.19) from Bacillus circulans strain 251 . Alignment of amino acid sequences of CGTases and alpha-amylases, and the analysis of the binding mode of the substrate analogue acarbose in the active site cleft {Strokopytov, B., et al . (1995) Biochemistry 34, (in press)}, suggested that Tyr195 plays an important role in cyclization of oligosaccharides . Tyr195 therefore was replaced with Phe (Y195F), Trp (Y195W), Leu (Y195L), and Gly (Y195G) . Mutant proteins were purified and crystallized, and their X-ray structures were determined at 2.5-2.6 angstrum resolution, allowing a detailed comparison of their biochemical properties and three-dimensional structures with those of the wild-type CGTase protein . The mutant proteins possessed significantly reduced cyclodextrin forming and coupling activities but were not negatively affected in the disproportionation and saccharifying reactions . Also under production process conditions, after a 45 h incubation with a 10% starch solution, the Y195W, Y195L, and Y195G mutants showed a lower overall conversion of starch into cyclodextrins . These mutants produced a considerable amount of linear maltooligosaccharides . The presence of aromatic amino acids (Tyr or Phe) at the Tyr195 position thus appears to be of crucial importance for an efficient cyclization reaction, virtually preventing the formation of linear products . Mass spectrometry of the Y195L reaction mixture, but not that of the other mutants and the wild type, revealed a shift toward the synthesis (in low yields) of larger products, especially of beta- and gamma- (but no alpha-) cyclodextrins and minor amounts of delta-, epsilon-, zeta- and eta-cyclodextrins.(ABSTRACT TRUNCATED AT 250 WORDS)

J Biol Chem, 1995 Mar 10, 270(10), 5490 - 4
Cloning and expression of a receptor for an insecticidal toxin of Bacillus thuringiensis; Vadlamudi RK et al.; Environmentally friendly toxins of Bacillus thuringiensis are effective in controlling agriculturally and biomedically harmful insects . However, little is known about the insect receptor molecules that bind these toxins and the mechanism of insecticidal activity . We report here for the first time the cloning and expression of a cDNA that encodes a receptor (BT-R1) of the tobacco hornworm Manduca sexta for an insecticidal toxin of B . thuringiensis . The receptor is a 210-kDa membrane glycoprotein that specifically binds the cryIA(b) toxin of B . thuringiensis subsp . berliner and leads to death of the hornworm . BT-R1 shares sequence similarity with the cadherin superfamily of proteins.

FEBS Lett, 1995 Mar 6, 360(3), 217 - 22
Delta-endotoxins induce cation channels in Spodoptera frugiperda brush border membranes in suspension and in planar lipid bilayers; Lorence A et al.; Membrane potential measurements using a fluorescent dye indicated that two specific toxins active against Spodoptera frugiperda larvae (CryIC and CryID) cause immediate permeability changes in midgut epithelial brush border membrane vesicles (BBMV) . The initial response and the sustained permeability change are cationic, not very K+ selective, and occur at in vivo lethal doses (nM) . The toxin response has a different ion selectivity and is more sensitive to Ba2+ than the intrinsic cation permeability of BBMV . Experiments incorporating BBMV into planar lipid bilayers (PLB) demonstrated that these vesicles contain cation channels (31, 47 and 76 pS) . A 2-40 fold conductance increase was induced by nM concentrations of toxin in PLB containing BBMV . Cationic single channel transitions of 50, 106, 360 and 752 pS were resolved . Thus, Bacillus thuringiensis delta-endotoxins induce an increase in cation membrane permeability involving ion channels in BBMV-containing functional receptors.

J Mol Biol, 1995 Mar 3, 246(4), 545 - 59
Crystal structure of calcium-depleted Bacillus licheniformis alpha-amylase at 2.2 A resolution; Machius M et al.; The three-dimensional structure of the calcium-free form of Bacillus licheniformis alpha-amylase (BLA) has been determined by multiple isomorphous replacement in a crystal of space group P4(3)2(1)2 (a = b = 119.6 A, c = 85.4 A) . The structure was refined using restrained crystallographic refinement to an R-factor of 0.177 for 28,147 independent reflections with intensities FObs > 0 at 2.2 A resolution, with root mean square deviations of 0.008 A and 1.4 degrees from ideal bond lengths and bond angles, respectively . The final model contains 469 residue, 237 water molecules, and one chloride ion . The segment between Trp182 and Asn192 could not be located in the electron density, nor could the N and C termini . Cleavage of the calcium-free form of BLA was observed after Glu189, due to a Glu-C endopeptidase present in trace amounts in the preparation . BLA did not crystallize without this cleavage under the conditions applied . BLA exhibits the characteristic overall topological fold observed for other alpha-amylases and related amylolytic enzymes: a central domain A containing an alpha/beta-barrel with a large protrusion between beta-strand 3 and alpha-helix 3 (domain B) and a C-terminal greek key motif (domain C) . Unlike in the other enzymes, domain B possesses a beta-sheet made up of six loosely connected, twisted beta-strands forming a kind of a barrel with a large hole in the interior . Topological comparisons to TAKA-amylase, pig pancreatic alpha-amylase and cyclodextrin glycosyltransferase reveal a very high structural equivalence for large portions of the proteins and an exceptionally pronounced structural similarity for calcium binding, chloride binding and the active site . None of the theories proposed to explain the enhanced thermostability of BLA showed a satisfactory correlation with the three-dimensional structure . Instead, sequence comparisons to the less thermostable bacterial alpha-amylase from Bacillus amyloliquefaciens (BAA) indicate that some ionic interactions present in BLA, but which cannot be formed in BAA, might be responsible for the enhanced thermostability of BLA.

J Mol Biol, 1995 Mar 3, 246(4), 511 - 21
The crystal structure of holo-glyceraldehyde-3-phosphate dehydrogenase from the hyperthermophilic bacterium Thermotoga maritima at 2.5 A resolution; Korndorfer I et al.; The crystal structure of holo-glyceraldehyde-3-phosphate dehydrogenase from the hyperthermophile Thermotoga maritima was determined by Patterson search methods using the known structure of the Bacillus stearothermophilus enzyme . The structure was refined at a resolution of 2.5 A to an R-factor of 16.63% for 26289 reflections between 8.0 A an 2.5 A with F > 2 sigma(F) . The crystallographic asymmetric unit contains two monomers related by approximate 2-fold symmetry and a tetramer is built up by crystallographic symmetry . The root-mean-square deviation of Ca positions of glyceraldehyde-3-phosphate dehydrogenase from T . maritima and B . stearothermophilus is 0.83 A in the NAD+ binding domains and smaller close to the cofactor . In contrast, the largest deviations in the catalytic domains are found at residues involved in coordination of sulphate ion SO4 339, which most likely marks the site of the attacking inorganic phosphate ion in catalysis . A large number of extra salt-bridges may be an important factor contributing to the high thermostability of this protein.

MMWR Morb Mortal Wkly Rep, 1995 Mar 3, 44(8), 137 - 40
Exposure of passengers and flight crew to Mycobacterium tuberculosis on commercial aircraft, 1992-1995; Effect of inorganic salts et al.; Vector Control Research Centre, Indian Council of Medical Research, Pondicherry, IndiaVarious inorganic salts and commonly used soaps and detergents were tested in the laboratory for their effect on the dissolution and larvicidal residual activity of a slow-release alginate encapsulated granular formation of Bacillus sphaericus . Fluoride, chloride and sulphate salts and a detergent powder affected the residual activity of this formulation drastically by rupturing it but did not effect its larvicidal activity . Nitrates and phosphates of sodium and potassium also had the same effect but to a moderate level . The safest concentration of these water impurities for effective functioning of the alginate encapsulated B . sphaericus formulation have been determined.

Am J Gastroenterol, 1995 Mar, 90(3), 485 - 8
Malignant histiocytosis in a patient presenting with hepatic dysfunction and peliosis hepatis; Fine KD et al.; In this article, we report the case of a 36-yr-old patient presenting with manifestations of portal hypertension, hepatic dysfunction, and fever who proved to have peliosis hepatis on liver biopsy . A thorough work-up revealed no obvious etiology . At autopsy, malignant histiocytosis of the liver and bone marrow was diagnosed . This case represents the first report of the association of peliosis hepatis with this rare histiocytic neoplasm and exemplifies the need for persistence in the search for malignancy, particularly hematological malignancy, in the patient with unexplained peliosis . The clinical similarity of peliosis hepatis associated with hematological malignancy and bacillary peliosis is also discussed.

Pediatrics, 1995 Mar, 95(3), 414 - 8
Bacillus Calmette-Guérin complications in children born to HIV-1-infected women with a review of the literature; O'Brien KL et al.; OBJECTIVE . To compare the risk of complications following Bacillus Calmette-Guerin (BCG) vaccination among children by maternal and infant HIV-1 infection status as part of an investigation of an outbreak of BCG complications . METHODS . A nonconcurrent cohort study of BCG complications among 125 infants born to HIV-1 seropositive and 166 infants born to HIV-1 seronegative mothers was conducted in Cite Soleil, Haiti . Infants were examined at regular intervals until 15 months of age, and complications from BCG were documented . An investigation of BCG vaccination practices was conducted . RESULTS . Mild or moderate complications occurred among 16 of 166 (9.6%) infants born to HIV-1 seronegative mothers compared with 4 of 13 HIV-1-infected infants (30.8%, P = .04) and 10 of 75 (13.3%, P = .39) uninfected infants born to HIV-1-infected mothers . No serious complications were noted . The outbreak of complications was associated with administration of 2.0 to 2.5 times the recommended dose of BCG vaccine . CONCLUSIONS . This and five other cohort studies indicate that there may be a small increased risk of complications following BCG vaccination among HIV-1-infected children, but the reactions are usually mild and the risk does not outweigh the benefits of BCG vaccination in populations at high risk of tuberculosis during infancy and childhood.

J Urol, 1995 Mar, 153(3 Pt 2), 929 - 33
A randomized study of intravesical mitomycin C, bacillus Calmette-Guerin Tice and bacillus Calmette-Guerin RIVM treatment in pTa-pT1 papillary carcinoma and carcinoma in situ of the bladder; Vegt PD et al.; Results of a randomized prospective study are reported in which mitomycin C, Tice bacillus Calmette-Guerin (BCG) and RIVM-BCG were compared in 437 patients with primary or recurrent pTa and pT1 bladder tumors, including carcinoma in situ . The followup (or time in study) varied from 2 to 81 months (mean 36 months) . After complete transurethral resection of all visible tumors the patients were treated with 30 mg . mitomycin C once a week for 4 consecutive weeks and thereafter every month for a total of 6 months, and 5 x 10(8) colony-forming units Tice BCG or RIVM-BCG once a week for 6 consecutive weeks . For papillary tumors mitomycin C and RIVM-BCG treatments were equally effective (p = 0.53), and mitomycin C was more effective than Tice BCG therapy (p = 0.01).

Appl Environ Microbiol, 1995 Mar, 61(3), 959 - 65
Characterization and substrate specificity of an endo-beta-1,4-D-glucanase I (Avicelase I) from an extracellular multienzyme complex of Bacillus circulans; Kim CH; An endo-1,4-beta-D-glucanase I (Avicelase I; EC 3.2.1.4) was purified to homogeneity from an extracellular celluloxylanosome of Bacillus circulans F-2 . The purification in the presence of 6 M urea yielded homogeneous enzyme . The enzyme had a monomeric structure, its relative molecular mass being 75 kDa as determined by gel filtration and 82 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . The pI was 5.4, and the N-terminal amino acid sequence was ASNIGGWVGGNESGFEFG . The optimal pH was 4.5, and the enzyme was stable at pH 4 to 10 . The enzyme has a temperature optimum of 50 degrees C, it was stable at 55 degrees C for 46 h, and it retains approximately 20% of its activity after 30 min at 80 degrees C . It showed high-level activity towards carboxymethyl cellulose (CMC) as well as p-nitrophenyl-beta-D-cellobioside, 4-methylumbelliferyl cellobioside, xylan, Avicel, filter paper, and some cello-oligosaccharides . Km values for birch xylan, CMC, and Avicel were 4.8, 7.2, and 87.0 mg/ml, respectively, while Vmax values were 256, 210, and 8.6 mumol x min-1 x mg-1, respectively . Cellotetraose was preferentially cleaved into cellobiose (G2) plus G2, and cellopentaose was cleaved into G2 plus cellotriose (G3), while cellohexaose was cleaved into cellotetraose plus G2 and to a lesser extent G3 plus G3 . G3 was not cleaved at all . G2 was the main product of Avicel hydrolysis . Xylotetraose (X4) and xylobiose (X2) were mainly produced by the enzyme hydrolysis of xylan . G2 inhibited the activity of carboxymethyl cellulase and Avicelase, whereas Mg2+ stimulated it . The enzyme was completely inactivated by Hg2+, and it was inhibited by a thiol-blocking reagent . Hydrolysis of CMC took place, with a rapid decrease in viscosity but a slow liberation of reducing sugars . On the basis of these results, it appeared that the cellulase should be regarded as endo-type cellulase, although it hydrolyzed Avicel.

Appl Environ Microbiol, 1995 Mar, 61(3), 941 - 3
Isolation and characterization of a cytotoxic metabolite of Talaromyces bacillosporus; Ishii K et al.; A cytotoxic metabolite, talarotoxin, was isolated from a fungus, Talaromyces bacillosporus IFO 8397, cultured on rice . The structure of the toxin was elucidated and found to contain a pyrrolizidinedione connected with a trans delta 1-octalin through a conjugated triene.

Appl Environ Microbiol, 1995 Mar, 61(3), 855 - 9
Stabilization of microbial cytochrome P-450 activity by creation of station-phase conditions in a continuously operated immobilized-cell reactor; Dror Y et al.; Bacillus megaterium (ATCC 13368) exhibits cytochrome P-450 monooxygenase activity (referred to herein as Cyt P-450 meg) catalyzing 15 beta-steroid hydroxylation . This activity belongs to the widespread ferredoxin reductase-ferredoxin-Cyt P-450 type of monooxygenases, providing a representative model system for this type of activity . The level of Cyt P-450 meg activity reaches its maximum in the cells during the stationary phase of the growth curve and is not affected by Cyt P-450 inducers . Here we present the development of an approach for stabilizing the Cyt P-450 meg system so that it performs continuous steroid hydroxylation and will be a model system for Cyt P-450-based detoxification . It is based on cell immobilization and simulation of stationary-phase conditions in a continuously operated fluidized-bed bioreactor . The combination of an appropriate immobilization technique, operational conditions, and medium composition provided a stabilized cell environment resulting in "freezing" of a physiological steady-state analog under stationary phase conditions, allowing stable performance of continuous hydroxylation for several weeks . It is suggested that this approach may be extended for use with other environmentally induced enzymatic activities.

Sante, 1995 Mar-Apr, 5(2), 89 - 94
{The recycling of waste water and mosquitoes}; Karch S et al.; Recycling waste water in the Acheres complex (North-West Paris) is based on both sophisticated industrial techniques and simple agricultural methods . The sewage farms and settling pools provide suitable breeding sites for more than ten mosquito species . Aedes caspius is the major pest for the local population . Moreover Culex pipiens (anthropophilic form) breeds in the sewers of the neighbouring towns . Mosquito control is based early ground treatment of breeding sites . Temephos and fenotrothion are used against A . caspius . Spherimos (Bacillus sphaericus) is used to control C . pipiens . The two insecticides have no adverse effect on humans or the environment and Spherimos is harmless . Aedes pest have been virtually eliminated . Urban Culex control is generally good despite being performed by less well trained municipal employees . In the areas treated by the specialized team of the SIAAP, pest mosquitoes have disappeared . The techniques used in the Acheres complex do not require sophisticated equipment . Thus, if adapted to local ecological, epidemiological and financial conditions they could be transferred to developing countries.

Biosci Biotechnol Biochem, 1995 Mar, 59(3), 529 - 31
The action of Bacillus circulans WL-12 chitinases on partially N-acetylated chitosan; Mitsutomi M et al.; Both chitinase A1 and D from Bacillus circulans WL-12 specifically hydrolyzed the N-acetyl-beta-D-glucosaminidic bonds in 50% N-acetylated chitosan molecules to produce hetero-oligosaccharides with GlcNAc at the reducing end residues, together with GlcNAc and (GlcNAc)2 . GlcN-GlcNAc and GlcN-GlcNAc-GlcNAc were produced as major hydrolysis products with chitinase A1 and D, respectively, but GlcN-GlcNAc was not detected in the digest of 50% N-acetylated chitosan with chitinase D.

Biotechnol Prog, 1995 Mar-Apr, 11(2), 231 - 4
Penicillin-G enhanced production of thuringiensin by Bacillus thuringiensis sp . darmstadiensis; Tzeng YM et al.; The effect of penicillin-G on the production of the potential microbial insecticide thuringiensin by Bacillus thuringiensis sp . darmstadiensis was studied . Shake flask and 3-L jar fermentor studies showed that the addition of 360 units/mL penicillin-G at 9 h, when the fermentable sugar in the medium was about to be mostly consumed, improved thuringiensin production by more than 1-fold relative to the control . The dosage of 360 units/mL penicillin-G had only a modest effect on the growth of the microorganism . However, cell growth was inhibited at higher dosages of the antibiotic . Since penicillin-G could interfere with cell wall synthesis, which facilitated the release of thuringiensin, a high thuringiensin productivity of 2600 mg/L was attained in this study, which is about 2-10-fold higher than those values reported in the literature.

Appl Microbiol Biotechnol, 1995 Mar, 42(6), 878 - 83
One-step enzymatic hydrolysis of starch using a recombinant strain of Saccharomyces cerevisiae producing alpha-amylase, glucoamylase and pullulanase; Janse BJ et al.; A recombinant strain of Saccharomyces cerevisiae was constructed that contained the genes encoding a bacterial alpha-amylase (AMY1), a yeast glucoamylase (STA2) and a bacterial pullulanase (pulA) . The Bacillus amyloliquefaciens alpha-amylase and S . cerevisiae var . diastaticus glucoamylase genes were expressed in S . cerevisiae using their native promoters and the encoded enzymes secreted under direction of their native leader sequences . In contrast, the Klebsiella pneumoniae pullulanase gene was placed under the control of the yeast alcohol dehydrogenase gene promoter (ADC1P) and secreted using the yeast mating pheromone alpha-factor secretion signal (MF alpha 1S) . Transcription termination of the pullulanase gene was effected by the yeast tryptophan synthase gene terminator (TRP5T), whereas termination of the glucoamylase and alpha-amylase genes was directed by their native terminators . Pullulanase (PUL1) produced by recombinant yeasts containing ADC1P MF alpha 1S pulA TRP5T (designated PUL1) was further characterized and compared to its bacterial counterpart (PulA) . The different genes were introduced into S . cerevisiae in different combinations and the various amylolytic Saccharomyces transformants compared to Schwanniomyces occidentalis . Introduction of PUL1 into a S . cerevisiae strain containing both STA2 and AMY1, resulted in 99% assimilation of starch.

Appl Microbiol Biotechnol, 1995 Mar, 42(6), 871 - 7
Inactivation of the major extracellular protease from Bacillus megaterium DSM319 by gene replacement; Wittchen KD et al.; An efficient method for gene replacement in Bacillus megaterium was developed and used to inactivate the chromosomal neutral protease gene (nprM) from strain DSM319 . A temperature-dependant suicide vector was constructed to allow replacement of the normal chromosomal copy with an altered version of the nprM gene . One mutant B . megaterium MS941 was selected for further characterization . Measurement of extracellular protease activity from strain MS941 indicated the existence of an additional minor extracellular protease in B . megaterium . Inhibitor studies revealed that this minor protease, comprising only 1.4% of the wild-type total extracellular protease activities, is a serine-type enzyme.

Bull Pan Am Health Organ, 1995 Mar, 29(1), 37 - 58
Epidemiology of AIDS and tuberculosis; Garcia Garcia ML et al.; This article reviews literature on the epidemiology, pathogenicity, and control of HIV and Mycobacterium tuberculosis coinfection . Regarding pathogenicity, immune system deterioration makes HIV-infected people more likely to develop active tuberculosis on primary or secondary exposure to the bacillus or to suffer reactivation of latent infections, and to experience considerably higher rates of extrapulmonary manifestations, relapses, and death . Regarding epidemiology, as of 1990 there were an estimated 3 million people coinfected with HIV and M . tuberculosis, with some 300,000 active tuberculosis cases and 120,000-150,000 tuberculosis deaths occurring annually among those coinfected . Over 500,000 coinfected people are thought to reside in the Americas, over 400,000 of them in Latin America . In general, the impact of coinfection is evident . Relatively high and increasing prevalences of HIV infection have been detected among tuberculosis patients around the world, and tuberculosis has become a frequent complication of AIDS cases . Moreover, there is no longer any doubt that coinfection obstructs tuberculosis prevention and control . Among other things, it affects BCG vaccination policies, suggests the need to administer preventive chemoprophylaxis to HIV-infected individuals at high risk of harboring or contracting tuberculosis infections, and complicates both detection and treatment of active tuberculosis cases . The recent proliferation of M . tuberculosis strains resistant to multiple drugs, most notably in the United States, compounds the problem . Tuberculosis prevention and control are still technically and economically feasible . However, more must be done to establish surveillance programs with laboratory support . More research is needed to determine what case prevention measures are best-suited to current circumstances and the HIV/AIDS presence . More effective preventive treatment regimens that are well tolerated, well complied with, and do not pose the risk of multiresistance need to be devised . More health workers need to be trained to suspect tuberculosis and to conduct timely and appropriate tests confirming this diagnosis . And finally, more must be done to standardize the types and durations of the various curative treatment regimens employed.

Clin Infect Dis, 1995 Mar, 20(3), 629 - 33
Bacillus licheniformis bacteremia: five cases associated with indwelling central venous catheters; Blue SR et al.; Bacillus species are being more frequently recognized as pathogens in immunocompromised hosts or in patients with cancer and central venous catheters . Only nine cases of Bacillus licheniformis infection have been reported in the English-language literature since 1966 . In a retrospective study we describe six patients and 17 episodes of B . licheniformis bacteremia over a 5-year span . All six patients had either a Hickman or a Broviac catheter in place for more than 3 months . Five of the six patients had multiple clinically significant episodes of bacteremia due to B . licheniformis . The six patients ranged in age from 4 years to 62 years . Two patients had leukemia or lymphoma and three patients had solid tumors, but only one patient was neutropenic . No deaths were related to B . licheniformis bacteremia . B . licheniformis should be considered as a potential pathogen in immunocompromised patients, especially when bacteremia is associated with the presence of long-term central venous catheters . Mortality due to B . licheniformis bacteremia is low, but recurrent bacteremia due to this organism causes significant morbidity and usually necessitates removal of the catheter.

AIDS, 1995 Mar, 9(3), 243 - 51
Safety and immunogenicity of a V3 loop synthetic peptide conjugated to purified protein derivative in HIV-seronegative volunteers; Rubinstein A et al.; OBJECTIVES: To develop a peptide-based model for a preventive vaccine for HIV-1 infection . DESIGN: Phase I trial in HIV-1-seronegative volunteers . PARTICIPANTS: Adult healthy subjects HIV-1-antibody-seronegative in an enzyme-linked immunosorbent assay, screened for tuberculin {purified protein derivative (PPD)} reactivity with 2 tuberculin units PPD-administered intradermally . INTERVENTIONS: Submicrogram doses of a PPD conjugate with a peptide of the primary neutralizing domain (PND) of HIV-1MN (PPD-MN-PND) were administered intradermally to tuberculin skin-test-positive and -negative volunteers . RESULTS: Antibodies to the MN-PND were measured after two immunizations in 10 out of 11 PPD skin-test-positive volunteers . After the fourth immunization high-affinity antibodies were detected, which persisted for over 1 year . High titers of MN-PND-specific immunoglobulin (Ig) G and IgA were detected in the serum and saliva of all volunteers tested . Serum antibodies were cross-reactive with PND peptide from some other HIV-1 strains but neutralized only the HIV-1MN prototype . Human leukocyte antigen (HLA)-B7-restricted MN-PND-specific cytotoxic T lymphocytes (CTL) were also detected . CONCLUSIONS: The PPD-MN-PND vaccine at submicrogram doses is safe and immunogenic in PPD skin-test-positive healthy adult volunteers . Long lasting humoral immune responses in the serum and saliva were possibly accompanied by HLA-B7-restricted CTL responses . This is a vaccine prototype that can be rapidly and inexpensively modified to include other peptide epitopes . It is especially suitable for use in a worldwide multibillion Bacillus Calmette-Guerin (BCG)-primed or tuberculosis-exposed population at risk for HIV-1 infection.

J Clin Microbiol, 1995 Mar, 33(3), 636 - 40
Bacillus cereus phage typing as an epidemiological tool in outbreaks of food poisoning; Ahmed R et al.; Bacillus cereus is responsible for an increasing number of food poisoning cases . By using 12 bacteriophages isolated from sewage, a typing scheme for B . cereus isolates from outbreaks or sporadic cases of food poisoning was developed . The phages belonged to three morphotypes . Ten phages with contractile tails and icosahedral heads were members of the Myoviridae family, and two phages with noncontractile tails belonged to the Siphoviridae family . Phage 11 represented a new species . It had an isometric head and a very long contractile tail with long wavy tail fibers and was one of the largest viruses known . The vast majority of 166 B . cereus strains (161, or 97%) isolated from food poisoning cases were typeable . Of 146 strains isolated from 18 outbreaks, 142 (97%) could be divided into 17 phage types . A good correlation, on the order of 80 to 100%, between phage types of strains isolated from suspected foods and those of strains isolated from stools of symptomatic patients was observed . Most Bacillus thuringiensis strains were also typeable, providing further evidence of the close relatedness of B . cereus and B . thuringiensis . This phage typing scheme can be a valuable epidemiological tool in tracing the origins of food poisoning caused by B . cereus.

J Antibiot (Tokyo), 1995 Mar, 48(3), 226 - 32
Inhibition of the binding of oxidized low density lipoprotein to the macrophages by iturin C-related compounds; Park JK et al.; Binding of modified lipoproteins including oxidized low density lipoprotein (oxidized LDL) to cell surface receptors is an initial step of conversion of monocyte-derived macrophages into lipid-laden foam cells, a key cellular component in the early lesions of atherosclerosis . We have searched for microbial metabolites that inhibit oxidized LDL-induced lipid accumulation in macrophages and isolated three compounds from a strain of Bacillus sp . as inhibitors of oxidized LDL binding . By chemical and spectroscopic analyses, these metabolites were shown to be related to the cyclic lipopeptide iturin C . Two of these compounds were novel metabolites having long chain beta-amino acid moieties of different length . These agents, at concentrations ranging from 5 to 20 microM, inhibited cell surface binding of oxidized 125I-LDL, resulting in reduced intracellular accumulation and degradation of the lipoprotein as well as in reduced cholesteryl ester formation from {14C}oleate in macrophages J774.

J Invertebr Pathol, 1995 Mar, 65(2), 162 - 73
The insecticidal CryIB crystal protein of Bacillus thuringiensis ssp . thuringiensis has dual specificity to coleopteran and lepidopteran larvae; Bradley D et al.; The crystals found in sporulation extracts of Bacillus thuringiensis (Berliner) contain proteins that are highly toxic to insects . Different crystal proteins exhibit distinct specificities for restricted groups of insects . An uncharacterized strain of B . thuringiensis (BtS2), derived from China, was found to carry several crystal protein genes and to be toxic to a wide variety of insects, including some coleopterans . Surprisingly, the coleopteran toxicity was traced to a CryIB-class protein . The previously cloned CryIB protein from B . thuringiensis ssp . thuringiensis strain HD-290-I, which was believed to be lepidopteran-specific, was also found to be toxic to at least two species of coleopteran larvae under certain conditions . In contrast to CryIB toxicity toward lepidopterans, the coleopteran activity of CryIB is enhanced by solubilization and by truncation with trypsin prior to administration . The magnitude of this effect varies with the host species and is reversed for the one lepidopteran tested . These results suggest that, for at least some insects, the apparent host specificity of CryIB may depend both on differences in midgut environment and on differences in toxin-receptor interaction . The results of insect toxicity experiments with a series of deletion mutants allowed definition of a CryIB protein fragment of ca . 65 kDa as the smallest peptide that retains bioactivity against both lepidopteran and coleopteran larvae . Deletions smaller than this resulted in the production of a protein that was nontoxic to both lepidopteran and coleopteran larvae.

Microbiology, 1995 Mar, 141 ( Pt 3), 649 - 54
Effects of signal peptide mutations on processing of Bacillus stearothermophilus alpha-amylase in Escherichia coli; Suominen I et al.; Bacillus stearothermophilus alpha-amylase has a signal peptide typical for proteins exported by Gram-positive bacteria . There is only one signal peptidase processing site when the protein is exported from the original host, but when it is exported by Escherichia coli, two alternative sites are utilized . Site-directed mutagenesis was used to study the processing in E . coli . Processing sites for 13 B . stearothermophilus alpha-amylases carrying mutations in their signal peptide were determined . Processing of the signal peptide was remarkably tolerant to mutations, because switching between the alternative sites was possible . The length and the sequence of the region between the hydrophobic core and the cleavage site was crucial for determining the choice of the processing site . Some mutations more distal to the cleavage site also affected the site preference.

Microbiology, 1995 Mar, 141 ( Pt 3), 629 - 39
Bacillus thuringiensis protoxin: location of toxic border and requirement of non-toxic domain for high-level in vivo production of active toxin; Wabiko H et al.; Insecticidal crystal proteins, or protoxins, of Bacillus thuringiensis are composed of two domains, an amino-terminal half essential for toxicity, and a carboxy-terminal half with an as yet unassigned function . To define the boundary of the two domains, sequential termination codons were introduced from the 3'-end of the DNA sequence encoding the toxic domain of the 1155-residue cry1A(b) gene product . The mutated and the intact genes were placed under the control of the Escherichia coli inducible promoter PrecA, and toxicity of the cell extracts was determined using silkworm larvae . Under non-induced conditions, in which the gene products accumulated to a limited degree, mutations encoding 606 amino acid residues or more were toxic, whereas those encoding 605 residues or less were non-toxic . Comparison of the toxicities and the levels of the toxic proteins suggested that the mutant proteins had comparable activity to that of the intact protoxin . Furthermore, the non-toxic protein seemed to be unstable in the extracts . To investigate the roles of the non-toxic domain, the mutant proteins were overproduced in both E . coli and B . thuringiensis . The intact and the mutated genes carrying natural promoters were introduced into acrystalliferous B . thuringiensis . Upon induction of PrecA in E . coli, and upon sporulation in B . thuringiensis, there was a large accumulation of gene products which formed inclusion bodies . The inclusion bodies of the intact protoxin were active, whereas those of the mutant proteins were inactive . Inclusion bodies of the intact protein could be solubilized in alkali, whereas the mutant inclusion bodies were insoluble . Since solubilization under alkaline conditions in the insect midgut is considered to be the first step of toxic action, the non-toxic domain is required to direct the synthesis of inclusion bodies as an active soluble form.

Eur J Immunol, 1995 Mar, 25(3), 838 - 46
Contribution of alpha/beta and gamma/delta T lymphocytes to immunity against Mycobacterium bovis bacillus Calmette Guérin: studies with T cell receptor-deficient mutant mice; Ladel CH et al.; Mutant mice with defined T cell deficiencies were infected with Mycobacterium bovis bacillus Calmette Guerin (BCG) and the relative contribution of alpha/beta T cells and gamma/delta T cells to the host immune response was assessed . Recombinase activating gene (RAG-1)-/- mutants as well as T cell receptor (TcR) beta-/-, but not TcR-delta-/-, mutants succumbed to M . bovis BCG infection and failed to develop granulomatous lesions . Antigen-induced IFN-gamma production by spleen cells in vitro was abrogated in RAG-1-/- mutants and markedly diminished in TcR-beta-/- and TcR-delta-/- mice . Reconstitution experiments suggest that both alpha/beta and gamma/delta T cells are essential for antigen-specific IFN-gamma secretion . Our data formally prove the crucial role of alpha/beta T cells and reveal accessory functions of gamma/delta T cells in optimum immunity against M . bovis BCG.

Eur J Immunol, 1995 Mar, 25(3), 672 - 6
Interleukin-12 is required for interferon-gamma production and lethality in lipopolysaccharide-induced shock in mice; Wysocka M et al.; Several cytokines, in particular tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma), have been shown to be responsible for pathological reactions which may lead to shock and death observed in infection with Gram-negative bacteria and in response to endotoxins (lipopolysaccharides, LPS) . Priming of mice with the avirulent Bacille Calmette Guerin (BCG) vaccine strain of Mycobacterium bovis increases the sensitivity of mice to the lethal effect of LPS and results in an efficient priming for cytokine production . In response to low doses (1 microgram/mouse) of LPS, BCG-primed mice produce interleukin-12 (IL-12) which controls IFN-gamma production, as demonstrated by the ability of neutralizing anti-IL-12 antibodies to suppress IFN-gamma production . However, the concentration of the biologically active IL-12 p70 heterodimer is similar in the serum of both BCG-primed or unprimed mice, reaching levels of 1-3 ng/ml at 3-6 h after LPS injection, whereas IFN-gamma production was observed only in BCG-primed mice . The priming effect of BCG on IFN-gamma production appears to be mostly due to its ability to increase TNF-alpha production, which acts as cofactor with LPS-induced IL-12 in inducing IFN-gamma production, as shown by the ability of injection of TNF-alpha and LPS (1 microgram/mouse), but not LPS alone, to induce IFN-gamma production . However, in addition to TNF-alpha, other LPS-induced cofactor(s) are required in cooperation with IL-12 to induce optimal IFN-gamma production, because co-injection of TNF-alpha and IL-12, sufficient to induce serum concentrations of both cytokines higher and more persistent than those obtained by injection of LPS, was not sufficient to induce IFN-gamma production in vivo . Neutralizing anti-IL-12 antibodies, in addition to inhibiting the in vivo LPS-induced IFN-gamma production, also completely protect BCG-primed mice injected with up to 10 micrograms of LPS from shock-induced death . Thus, IL-12 is required for IFN-gamma production and lethality in an endotoxic shock model in mice.

Br J Ophthalmol, 1995 Mar, 79(3), 250 - 6
Is leprosy blindness avoidable? The effect of disease type, duration, and treatment on eye damage from leprosy in Uganda; Waddell KM et al.; AIMS--The study was designed to measure the prevalence, range, and severity of eye involvement in leprosy patients; to relate this to disease type, duration, and treatment to identify risk factors; and to provide practical guidelines for programme managers and field staff on the prevention of blindness . METHODS--The visual outcome was assessed in a population based sample of patients in Kasese District, Uganda followed for up to two decades, and related to disease features and treatment . A total of 678 patients responded to an invitation out of 2715 registered since 1973 . RESULTS--Low vision was present in 4.4% of people and blindness in 1.3%, with 1.5% and 0.6% respectively being due to leprosy . Some 12.4% of patients had iritis, of whom 33% had visual loss in one or both eyes, 3.7% of patients had lagophthalmos, and 11.7% had lens opacity . For multi-bacillary (PB) cases, the adjusted odds ratios were: for iritis 4.6 (95% CI 2.6-8.2), for lagophthalmos 1.4 (0.6-3.2), and for lens opacity 1.7 (1.0-3.0) . Potentially sight threatening (PST) lesions were present in 16.8% of patients (95% CI 14.0-19.6) . CONCLUSION--Levels of eye involvement in this study are low compared with many surveys . Visual loss is uncommon and is more often caused by other diseases; in the present era of multidrug therapy (MDT) it is very unlikely to be caused by leprosy . It is more common with advancing age . PST lesions, especially iritis, may occur in both PB and MB cases, even if the diagnosis of leprosy is made early and MDT started immediately; they may occur also after completion of MDT . But eye complications need not proceed to loss of sight if treated promptly, and blindness can be avoided . Training of front line staff is therefore crucial.

J Biochem (Tokyo), 1995 Mar, 117(3), 467 - 70
Homogeneity of the pyruvate dehydrogenase multienzyme complex from Bacillus stearothermophilus; Hiromasa Y et al.; The pyruvate dehydrogenase multienzyme complex was purified from Bacillus stearothermophilus by means of six gel-filtration column chromatographies; once on Cellulofine GCL-2000, twice on Sepharose CL-2B, and three times on Sephacryl S-500HR . The molecular size distribution of the complex was examined in detail by gel-filtration chromatography, analytical and sucrose-density ultracentrifugations, and dynamic light scattering . The complex was found to be homogeneous; a dimeric complex was undetectable even with a high concentration of protein (below 6.8 mg/ml).

Mol Microbiol, 1995 Mar, 15(6), 1049 - 53
Protein kinase-dependent HPr/CcpA interaction links glycolytic activity to carbon catabolite repression in gram-positive bacteria; Deutscher J et al.; CcpA, the repressor/activator mediating carbon catabolite repression and glucose activation in many Gram-positive bacteria, has been purified from Bacillus megaterium after fusing it to a His tag . CcpA-his immobilized on a Ni-NTA resin specifically interacted with HPr phosphorylated at seryl residue 46 . HPr, a phospho-carrier protein of the phosphoenolpyruvate: glycose phosphotransferase system (PTS), can be phosphorylated at two different sites: (i) at His-15 in a PEP-dependent reaction catalysed by enzyme I of the PTS; and (ii) at Ser-46 in an ATP-dependent reaction catalysed by a metabolite-activated protein kinase . Neither unphosphorylated HPr nor HPr phosphorylated at His-15 nor the doubly phosphorylated HPr bound to CcpA . The interaction with seryl-phosphorylated HPr required the presence of fructose 1,6-bisphosphate . These findings suggest that carbon catabolite repression in Gram-positive bacteria is a protein kinase-triggered mechanism . Glycolytic intermediates, stimulating the corresponding protein kinase and the P-ser-HPr/CcpA complex formation, provide a link between glycolytic activity and carbon catabolite repression . The sensitivity of this complex formation to phosphorylation of HPr at His-15 also suggests a link between carbon catabolite repression and PTS transport activity.

Gastroenterol Hepatol, 1995 Mar, 18(3), 125 - 8
{Gastric tuberculosis: a report of 2 cases}; Fernandez de la Puebla Gimenez RA et al.; Two patients with gastric tuberculosis are described . The first presented toxic syndrome, multiple abdominal adenopathies, microcytic anemia and a subcardial ulcer with malignant characteristics at endoscopy . Diagnosis was based on the positivity of Ziehl-Neelsen staining and on the growth of Mycobacterium tuberculosis in the culture of the gastric mucosa . The second patient presented toxic syndrome, fever and a miliary pattern on thoracic radiography . Endoscopy demonstrated an ulcerated nodular lesion with granulomas with acid alcohol resistant bacillus being observed on biopsy . Mycobacterium tuberculosis was found in both the sputum and bronchoaspirate . The evolution of both cases was favorable with specific treatment.

J Am Mosq Control Assoc, 1995 Mar, 11(1), 86 - 9
Control of Anopheles stephensi breeding in construction sites and abandoned overhead tanks with Bacillus thuringiensis var . israelensis; Kumar A et al.; Bacillus thuringiensis (H-14), strain 164 (Bactoculicide) when applied at 1 g/m2 surface area successfully controlled Anopheles stephensi breeding in construction sites, abandoned overhead tanks, and curing waters . Subsequent to application, no pupal production was observed in these habitats for 3, 18, and 21 days, respectively . Based on these findings, inclusion of Bactoculicide in the bioenvironmental vector control strategy is suggested and fortnightly spraying in construction sites at 1 g/m2 surface area is recommended for the containment of vector breeding . However, frequent retreatment of abandoned overhead tanks would be uneconomical and operationally impractical.

J Am Mosq Control Assoc, 1995 Mar, 11(1), 72 - 6
Comparative toxicity of selected larvicides and insect growth regulators to a Florida laboratory population of Aedes albopictus; Ali A et al.; Five organophosphates (OPs) (chlorpyrifos, chlorpyrifos methyl, fenthion, malathion, and temephos), 3 pyrethroids (bifenthrin, cypermethrin, and permethrin), and 2 microbial pesticides (Bacillus thuringiensis serovar.israelensis {B.t.i.} and Bacillus sphaericus) were tested as larvicides against a Florida Aedes albopictus population colonized in the laboratory . In addition, 3 insect growth regulators (IGRS) (diflubenzuron, methoprene, and pyriproxyfen) were evaluated . All OPs, except for malathion, were highly effective as indicated by low LC90s ranging from 0.0069 ppm (chlorpyrifos) to 0.026 ppm (fenthion); the larvae were considered tolerant to malathion (LC90 = 1.043 ppm) . LC90 values of pyrethroids were: 0.0175 ppm (bifenthrin), 0.0079 ppm (cypermethrin), and 0.0031 ppm (permethrin) . Commercial products of B.t.i., Vectobac and Bactimos were considered economically effective against Ae . albopictus larvae but products of B . sphaericus were ineffective (LC90s > 28 ppm) . The IGRs showed exceptional activity . Pyriproxyfen (LC90 = 0.000376 ppm), was 2.23 and 21.5 times more toxic than diflubenzuron and methoprene, respectively . In general, toxicity ranking of chemicals and microbials tested was: IGRs > pyrethroids > OPs > microbials.

J Am Mosq Control Assoc, 1995 Mar, 11(1), 107 - 10
Effect of low temperature on feeding rate of Aedes stimulans larvae and efficacy of Bacillus thuringiensis var . israelensis (H-14); Walker ED; Experiments were conducted to determine the effects of low temperature (0 and 4 degrees C), vs . a high temperature (22 degrees C), on the feeding rate of Aedes stimulans larvae, and their susceptibility to Bacillus thuringiensis var . israelensis (H-14) (B.t.i.) . Third-instar Ae . stimulans slowed but did not halt feeding at 0 and 4 degrees C compared to 22 degrees C . Susceptibility of larvae, as measured by LC50 values, to B.t.i . was highest at 22 degrees C (LC50 = 0.1 ppm), and lower at 4 degrees C (LC50 = 0.2 ppm) and 0 degree C (LC50 = 0.9 ppm) . The data from the feeding and susceptibility experiments suggest that decreased efficacy of B.t.i . at low temperatures may occur because the rate of larval feeding decreases . Low water temperature should be a consideration during operational applications of B.t.i . for control of larvae in cold-water habitats, such as the spring Aedes species.

J Am Mosq Control Assoc, 1995 Mar, 11(1), 1 - 5
Development of a high level of resistance to Bacillus sphaericus in a field population of Culex quinquefasciatus from Kochi, India; Rao DR et al.; Field resistance to Bacillus sphaericus was observed in a population of Culex quinquefasciatus in Kochi, India, exposed to 35 rounds of spraying with a formulation of B . sphaericus 1593 M over a 2-year period . Larvae from the sprayed area gave LC50 and LC90 values that were 146 and 180 times greater than corresponding values for a susceptible strain from an unsprayed locality . When the resistant strain was colonized in the laboratory and subjected to moderate selection pressure at each generation, resistance rapidly increased and by the 18th generation it was 6,223 and 31,325 times greater at the LC50 and LC90 levels in comparison with the susceptible strain . There were no significant differences among 6 susceptible strains tested . Tests were repeated and validated using the standard primary powder SPH88, B . sphaericus 2362 . No cross resistance was observed against B . thuringiensis H-14.

J Mol Cell Cardiol, 1995 Mar, 27(3), 893 - 900
Phospholipid degradation in hypoxic/reoxygenated cardiomyocytes in response to phospholipase C from Bacillus cereus; Forsdahl K et al.; In the present study, we investigated possible mechanisms behind exogenous phospholipase C-induced glycerol production in irreversibly damaged myocytes . Rat ventricular myocytes were preincubated for 60 min in substrate-free Krebs-Henseleit bicarbonate buffer equilibrated with 95% N2-5% CO2 (37 degrees C, pH = 7.4), resulting in exhaustion of cellular high energy phosphates and loss of rod-shaped morphology . At the end of the preincubation period, the incubation vials were divided into two groups; one receiving 10 mU/ml phospholipase C (PC-PLC), whereas the other received an equivalent volume of buffer (control incubations) . Incubation was then continued for another 60 min under 95% air-5% CO2 atmosphere . Samples for measurement of metabolite levels were taken immediately after cell isolation, at the end of the preincubation period and at the end of the normoxic incubation period . During the 60 min incubation period following reoxygenation, glycerol output was markedly higher from PC-PLC treated than from control myocytes . However, the elevated glycerol output from these cells was not accompanied by a simultaneous rise in glycerol-3-phosphate, nor was it inhibited by inclusion of pyruvate in the incubation buffer . On the other hand, glycerol output from PC-PLC treated myocytes was effectively inhibited by a diacylglycerol lipase inhibitor (U-57908, The Upjohn Company) . Analysis of cellular lipids revealed a 22% reduction of phospholipid in PC-PLC treated myocytes (P < 0.02), while the content of triacylglycerol, diacylglycerol and unesterified fatty acids increased by 76, 261 and 103%, respectively (P < 0.02) . No significant changes were observed for these parameters in control myocytes.(ABSTRACT TRUNCATED AT 250 WORDS)

J Pharm Pharmacol, 1995 Mar, 47(3), 177 - 81
Interaction between fibronectin-bearing surfaces and Bacillus Calmette-Guérin (BCG) or gelatin microparticles; Lou Y et al.; Gelatin, prepared commercially by degradation of animal collagen, was studied to see whether it had an affinity for fibronectin, which has a known affinity for collagen, and whether gelatin-based drugs could be used to target fibronectin-excreting tumours . Bacillus Calmette-Guerin (BCG) vaccine, an attenuated strain of Mycobacterium bovis, is currently the most effective treatment for superficial transitional cell carcinoma of the bladder . The living cells of the BCG vaccine associate with the fibronectin-bearing surfaces of the tumour . Using a multi-well culture plate technique, gelatin microparticles were shown to be adsorbed onto murine S180 sarcoma cells and this reaction was substantially inhibited by the addition of human plasma fibronectin . The avidities of various BCG substrains and gelatin microparticles for glass-bound fibronectin were measured and the association constants determined . The gelatin microparticles associated with the fibronectin with equal avidity as the BCG cells . The results suggest that this model system may allow the investigation of gelatin-based drug delivery devices capable of targeting fibronectin-bearing surfaces associated with some tumours.

Dig Dis, 1995 Mar-Apr, 13(2), 108 - 18
Whipple's disease; Fantry GT et al.; Whipple's disease is a chronic systemic infectious disease caused by Tropheryma whippelii that typically involves the small intestine and causes malabsorption . Extraintestinal manifestations such as arthritis and fever are common and often exist prior to the onset of gastrointestinal symptoms . Involvement of the central nervous system can occur and lead to permanent sequelae . Weight loss, hyperpigmentation, and lymphadenopathy are frequent findings . The definitive diagnosis is made by biopsy of the small intestine mucosa which reveals infiltration of the lamina propria of the small intestine with periodic acid-Schiff positive macrophages . Treatment with trimethoprim combined with sulfamethoxazole for 1 year usually results in clinical remission and an excellent prognosis . Recent advances using molecular techniques to identify the uncultured bacillus of Whipple's disease should lead to a better understanding of the pathophysiology and allow for the development of a sensitive noninvasive diagnostic test.

J Clin Microbiol, 1995 Mar, 33(3), 742 - 4
A chemically defined liquid medium that supports primary isolation of Rochalimaea (Bartonella) henselae from blood and tissue specimens; Wong MT et al.; Rochalimaea (Bartonella) henselae is a fastidious, slowly growing, gram-negative bacillus that is an etiologic agent of bacillary angiomatosis, cat scratch disease, and related syndromes . Accumulation of direct microbiologic evidence of the relationship between the organism and the syndromes compatible with cat scratch disease has been hindered by the difficulties in the primary isolation of the organism from infected tissue specimens . A chemically defined liquid medium was developed to support the growth of Rochalimaea species to facilitate study of the organism . This medium was also used successfully to isolate R . henselae from clinical specimens from infected patients and a domestic cat . Recovery of R . henselae in this was more successful than when recovery was attempted on solid agar . This cell-free, extract-free, defined medium additionally supported the growth of Rochalimaea quintana and Afipia felis.

J Bacteriol, 1995 Mar, 177(6), 1444 - 51
The S-layer from Bacillus stearothermophilus DSM 2358 functions as an adhesion site for a high-molecular-weight amylase; Egelseer E et al.; The S-layer lattice from Bacillus stearothermophilus DSM 2358 completely covers the cell surface and exhibits oblique symmetry . During growth of B . stearothermophilus DSM 2358 on starch medium, three amylases with molecular weights of 58,000, 98,000, and 184,000 were secreted into the culture fluid, but only the high-molecular-weight enzyme was found to be cell associated . Studies of interactions between cell wall components and amylases revealed no affinity of the high-molecular-weight amylase to isolated peptidoglycan . On the other hand, this enzyme was always found to be associated with S-layer self-assembly products or S-layer fragments released during preparation of spheroplasts by treatment of whole cells with lysozyme . The molar ratio of S-layer subunits to the bound amylase was approximately 8:1, which corresponded to one enzyme molecule per four morphological subunits . Immunoblotting experiments with polyclonal antisera against the high-molecular-weight amylase revealed a strong immunological signal in response to the enzyme but no cross-reaction with the S-layer protein or the smaller amylases . Immunogold labeling of whole cells with anti-amylase antiserum showed that the high-molecular-weight amylase is located on the outer face of the S-layer lattice . Because extraction of the amylase was possible without disintegration of the S-layer lattice into its constituent subunits, it can be excluded that the enzyme is incorporated into the crystal lattice and participates in the self-assembly process . Affinity experiments strongly suggest the presence of a specific recognition mechanism between the amylase molecules and S-layer protein domains either exposed on the outermost surface or inside the pores . In summary, results obtained in this study confirmed that the S-layer protein from B . stearothermophilus DSM 2358 functions as an adhesion site for a high-molecular-weight amylase.

Biochemistry, 1995 Feb 28, 34(8), 2560 - 5
Role of glycine 212 in the allosteric behavior of phosphofructokinase from Bacillus stearothermophilus; Zhu X et al.; Crystallographic studies indicate that the loop between alpha-helix 8 and beta-strand H (the 8H loop) which borders the effector site of Bacillus stearothermophilus phosphofructokinase (BsPFK) is involved in the allosteric mechanism of the enzyme {Schirmer, T., and Evans, P.R . (1990) Nature 343, 140-145} . The residue at one end of this loop, glycine 212, has been proposed to be a pivot about which the loop hinges . Using site-directed mutagenesis, glycine 212 was replaced with valine (G212V) . Steady-state kinetic analysis and ligand binding studies on the altered and native PFKs showed that the G212V substitution resulted in discernible changes at the effector site . The mutated PFK required a 3-fold higher concentration of the allosteric inhibitor phosphoenolpyruvate than did the native enzyme to cause the same level of inhibition . The altered PFK had a 2-fold higher dissociation constant for the allosteric activator GDP than the wild-type enzyme . More importantly, whereas the native PFK was fully activated by 1 mM GDP from its PEP-inhibited T-state, the altered enzyme was only marginally activated . On the other hand, the G212V mutation resulted in no changes at the catalytic site of BsPFK . The catalytic rate constant kcat remained unchanged . The altered PFK had the same Km values for ATP and fructose-6-phosphate (Fru-6-P) as did the wild-type enzyme . Furthermore, starting from the same PEP-inhibited T-state, both enzymes gave identical sigmoidal responses to increasing Fru-6-P concentration, indicating that Fru-6-P can activate both to the R-state.

J Biol Chem, 1995 Feb 24, 270(8), 3828 - 35
A chimeric bacterial phosphofructokinase exhibits cooperativity in the absence of heterotropic regulation; Byrnes WM et al.; The phosphofructokinases (PFKs) from the bacteria Escherichia coli and Bacillus stearothermophilus differ markedly in their regulation by ATP . Whereas E . coli PFK (EcPFK) is profoundly inhibited by ATP, B . stearothermophilus PFK (BsPFK) is only slightly inhibited . The structural basis for this difference could be closure of the active site via a conformational transition that occurs in the ATP-binding domain of EcPFK, but is absent in BsPFK . To investigate the role of this transition in ATP inhibition of EcPFK, we have constructed a chimeric enzyme that contains the "rigid" ATP-binding domain of BsPFK grafted onto the remainder of the EcPFK subunit . The chimeric PFK has the following characteristics: (i) tetrameric structure and kinetic parameters similar to those of the native enzymes, (ii) insensitivity to regulation by the effector phosphoenolpyruvate despite its ability to bind to the enzyme, and (iii) a sigmoidal (nH around 2) fructose 6-phosphate saturation curve . From the results, it is concluded that the active site regions of the two native enzymes are remarkably similar, but their effector sites and their mechanisms of heterotropic regulation are different . The chimeric subunit is locked in a structure resembling that of activated E . coli PFK, yet the enzyme can exist in two different conformational states . Mechanisms for its sigmoidal kinetics are discussed.

Biochim Biophys Acta, 1995 Feb 22, 1247(1), 97 - 103
The role of histidine residues in the catalytic act of cyclomaltodextrin glucanotransferase from Bacillus circulans var . alkalophilus; Mattsson P et al.; Our previous study on cyclomaltodextrin glucanotransferase (CGTase) by chemical modification implied the importance of one or two histidine residues in the cyclization reaction of the enzyme . Based on a computer modelled three-dimensional structure of the CGTase, five histidine residues were chosen as targets for the site-directed mutagenesis . The histidine residues 98, 140, 233 and 327 were replaced by aspartate and His-177 by proline using polymerase chain reaction-mediated techniques . The CGTase variants H98D, H140D, H233D and H327D resulted in a profound decrease in the cyclizing and amylolytic activities, while mutation H177P had little influence on the activities but affected the thermal stability and the width of the pH optimum . It is suggested that His-98 functions as (or as a significant part of) the subsite 2 for the binding of the substrate in CGTase and therefore H98D destabilizes the intermediate for cyclization, but does not markedly affect the hydrolytic reactions . Mutants H140D and H233D produced only minor amounts of alpha-cyclodextrin, did not exhibit substrate inhibition with maltotriose and showed non-Michaelis-Menten kinetics . It is proposed that the variants H140D, H233D and H327D cause steric hindrances near the active center, while mutation H177D has similar consequences on the same site spatially.

Biochemistry, 1995 Feb 21, 34(7), 2234 - 40
X-ray structure of cyclodextrin glycosyltransferase complexed with acarbose . Implications for the catalytic mechanism of glycosidases; Strokopytov B et al.; Crystals of cyclodextrin glycosyltransferase (CGTase) from Bacillus circulans strain 251 were soaked in buffer solutions containing the pseudotetrasaccharide acarbose, a strong amylase- and CGTase inhibitor . The X-ray structure of the complex was elucidated at 2.5-A resolution with a final crystallographic R value of 15.8% for all data between 8.0 and 2.5 A . Acarbose is bound near the catalytic residues Asp229, Glu257, and Asp328 . The carboxylic group of Glu257 is at hydrogen bonding distance from the glycosidic oxygen in the scissile bond between the B and C sugars (residue A is at the nonreducing end of the inhibitor) . Asp328 makes hydrogen bonds with the 4-amino-4,6-dideoxyglucose (residue B), and Asp229 is in a close van der Waals contact with the C1 atom of this sugar . From this we conclude that in CGTase Glu257 acts as the proton donor and Asp229 serves as the general base or nucleophile, while Asp328 is involved in substrate binding and may be important for elevating the pKa of Glu257 . On the basis of these results it appears that the absence of the C6-hydroxyl group in the B sugar is responsible for the inhibitory properties of acarbose on CGTase . This suggests that the C6-hydroxyl group of this sugar plays an essential role in the catalytic mechanism of CGTase.(ABSTRACT TRUNCATED AT 250 WORDS)

J Biol Chem, 1995 Feb 17, 270(7), 3081 - 8
Crystal structure and site-directed mutagenesis of Bacillus macerans endo-1,3-1,4-beta-glucanase; Hahn M et al.; In beta-glucans those beta-1,4 glycosidic bonds which are adjacent to beta-1,3 bonds are cleaved by endo-1,3-1,4-beta-glucanases (beta-glucanases) . Here, the relationship between structure and activity of the beta-glucanase of Bacillus macerans is studied by x-ray crystallography and site-directed mutagenesis of active site residues . Crystal structure analysis at 2.3-A resolution reveals a jelly-roll protein structure with a deep active site channel harboring the amino acid residues Trp101, Glu103, Asp105, and Glu107 as in the hybrid Bacillus beta-glucanase H(A16-M) (Keitel, T., Simon, O., Borriss, R., and Heinemann, U . (1993) Proc . Natl . Acad . Sci . U.S.A . 90, 5287-5291) . Different mutant proteins with substitutions in these residues are generated by site-directed mutagenesis, isolated, and characterized . Compared with the wild-type enzyme their activity is reduced to less than 1% . Several mutants with isosteric substitutions in Glu103 and Glu107 are completely inactive, suggesting a direct role of these residues in glycosyl bond hydrolysis . The kinetic properties of mutant beta-glucanases and the crystal structure of the wild-type enzyme are consistent with a mechanism where Glu103 and Glu107 are the catalytic amino acid residues responsible for cleavage of the beta-1,4 glycosidic bond within the substrate molecule.

Eur J Biochem, 1995 Feb 15, 228(1), 206 - 10
Resistance in a laboratory population of Culex quinquefasciatus (Diptera: Culicidae) to Bacillus sphaericus binary toxin is due to a change in the receptor on midgut brush-border membranes; Nielsen-Leroux C et al.; Direct binding experiments with isolated brush border membrane fractions (BBMF) from larvae of a susceptible laboratory strain of Culex quinquefasciatus Say, indicated the presence of a single class of Bacillus sphaericus binary toxin receptors . The dissociation constant (Kd) was approximately 11 nM and the maximum binding capacity (Bmax) approximately 8 pmol/mg BBMF protein . Similar binding experiments with a field population of C . quinquefasciatus that had been selected in the laboratory to more than 100,000-fold resistance to B . sphaericus binary toxin failed to reveal the presence of any specific binding . Thus this resistant strain had lost the functional receptor for B . sphaericus toxin . The binding characteristics of BBMF from the F1 larval progeny (susceptible females x resistant males) were very close to those of the parental susceptible strain, consistent with the resistance being recessive.

FEMS Microbiol Lett, 1995 Feb 15, 126(2), 133 - 7
Characterization of a DNA fragment carrying the raw starch-digesting alpha-amylase and salt-dependent alpha-amylase genes from Bacillus circulans F-2; Kim CH; A 5.4 kb HindIII DNA fragment carrying the gene encoding raw starch-digesting alpha-amylase (RSDA), has been previously cloned from Bacillus circulans F-2 and expressed in Escherichia coli {Kim et al . (1990) Biochim . Biophys . Acta 1048, 2233-2238} . Interestingly, when the cell extract of E . coli harboring a plasmid carrying this fragment was incubated with 1 M NaCl, it exhibited about 10 times higher enzyme activity than when assayed without NaCl . Differential zymograms showed two different amylase activities: one for RSDA and the other for a salt-dependent alpha-amylase (SDA) . Even though RSDA activity was detected without NaCl, SDA activity was detected only in high concentrations of NaCl . SDA activity was fully detected at above 1 M NaCl . Results from subcloning of the genes, fractionation analysis of cell extracts, and immunological assays clearly suggested that the two amylases are genetically distinct and that genes for both enzymes are closely linked on the 5.4 kb DNA fragment.

Biochem Pharmacol, 1995 Feb 14, 49(4), 567 - 74
Activation and cytotoxicity of 2-alpha-aminoacyl prodrugs of methotrexate; Smal MA et al.; In an effort to improve the selectivity of the anticancer drug methotrexate (MTX), a series of potential prodrugs in which the 2-amino group was acylated with various alpha-amino acids (as well as L-pyroglutamic acid) was synthesized . Such derivatives are anticipated to be hydrolysed to MTX by appropriate aminopeptidases localized (over-expressed naturally or targeted as anti-tumor antibody conjugates) in the vicinity of the tumor . The L-leucyl, L-valyl, L-isoleucyl, D-alanyl and L-pyroglutamyl derivatives were assessed as to their suitability as prodrugs . Except for the L-pyroglutamyl compound, all derivatives decomposed slowly when incubated in phosphate buffer, pH 7.3; the formation of MTX was minimal . No major differences were observed when serum was included in the incubation medium, except for the L-leucyl compound, which was hydrolysed to MTX . The L-leucyl, L-valyl and L-isoleucyl derivatives were hydrolysed readily to MTX by aminopeptidase M (EC 3.4.11.2), while the L-pyroglutamyl and D-alanyl compounds were activated by pyroglutamate aminopeptidase (EC 3.4.19.3) (from Bacillus amyloliquefaciens) and D-aminopeptidase (from Ochrobactrum anthropi), respectively . When tested for inhibition of the target enzyme dihydrofolate reductase (DHFR; EC 1.5.1.3), 2-L-valyl-MTX showed inhibition two orders of magnitude poorer than that given by MTX, in agreement with the expectation that acylation of the 2-amino group reduces binding to DHFR . After treatment of this derivative with aminopeptidase M, the extent of inhibition correlated with the amount of MTX formed . MTX derivatives alone or in combination with the complementary peptidase were tested for cytotoxicity on murine L1210 cells in culture . The above-listed derivatives were considerably less cytotoxic than MTX, except for the L-leucyl derivative which showed considerable cytotoxicity . When the appropriate exogenous peptidase was included, the cytotoxicity of the activated prodrugs approached that of MTX . These results indicate that 2-L-leucyl-MTX is unsuitable as a prodrug since it is activated prematurely by serum enzymes . Although the L-valyl and L-isoleucyl derivatives do not hydrolyse to MTX in serum and are readily activated, they are not ideal prodrugs since they decompose under physiological conditions; the properties of the decomposition product will have a bearing on the ultimate suitability of these compounds . 2-L-Pyroglutamyl-MTX is the best candidate prodrug, showing stability and ready activation by the appropriate aminopeptidase.

J Biol Chem, 1995 Feb 10, 270(6), 2571 - 8
The assembly and organization of the alpha 5 and alpha 7 helices from the pore-forming domain of Bacillus thuringiensis delta-endotoxin . Relevance to a functional model; Gazit E et al.; The pore-forming domain of Bacillus thuringiensis insecticidal CryIIIA delta-endotoxin contains two helices, alpha 5 and alpha 7, that are highly conserved within all different Cry delta-endotoxins . To gain information on the mode of action of delta-endotoxins, we have used a spectrofluorimetric approach and characterized the structure, the organization state, and the ability to self-assemble and to co-assemble within lipid membranes of alpha 5 and alpha 7 . Circular dichroism (CD) spectroscopy revealed that alpha 7 adopts a predominantly alpha-helical structure in methanol, similar to what has been found for alpha 5, and consistent with its structure in the intact molecule . The hydrophobic moment of alpha 7 is higher than that calculated for alpha 5; however, alpha 7 has a lesser ability to permeate phospholipids as compared to alpha 5 . Binding experiments with 7-nitrobenz-2-oxa-1,3-diazole-4-yl (NBD)-labeled peptide demonstrated that alpha 7 binds to phospholipid vesicles with a partition coefficient in the order of 10(4) M-1 similar to alpha 5, but with reduced kinetics and in a noncooperative manner, as opposed to the fast kinetics and cooperativity found with alpha 5 . Resonance energy transfer measurements between fluorescently labeled pairs of donor (NBD)/acceptor (rhodamine) peptides revealed that, in their membrane-bound state, alpha 5 self-associates but alpha 7 does not, and that alpha 5 coassembles with alpha 7 but not with an unrelated membrane bound alpha-helical peptide . Furthermore, resonance energy transfer experiments, using alpha 5 segments, specifically labeled in either the N- or C-terminal sides, suggest a parallel organization of alpha 5 monomers within the membranes . Taken together the results are consistent with an umbrella model suggested for the pore forming activity of delta-endotoxin (Li, J., Caroll, J., and Ellar, D . J . (1991) Nature 353, 815-821), where alpha 5 has transmembrane localization and may be part of the pore lining segment(s) while alpha 7 may serve as a binding sensor that initiates the binding of the pore domain to the membrane.

J Biol Chem, 1995 Feb 10, 270(6), 2517 - 24
Glycan requirements of glycosylphosphatidylinositol phospholipase C from Trypanosoma brucei . Glucosaminylinositol derivatives inhibit phosphatidylinositol phospholipase C; Morris JC et al.; Glycosylphosphatidylinositol phospholipase C (GPI-PLC) from Trypanosoma brucei and phosphatidylinositol phospholipase C (PI-PLC) from Bacillus sp . both cleave glycosylphosphatidylinositols (GPIs) . However, phosphatidylinositol, which is efficiently cleaved by PI-PLC, is a very poor substrate for GPI-PLC . We examined GPI-PLC substrate requirements using glycoinositol analogs of GPI components as potential inhibitors . Glucosaminyl (alpha 1-->6)-D-myo-inositol (GlcN(alpha 1-->6)Ins), GlcN(alpha 1-->6)Ins 1,2-cyclic phosphate, GlcN(alpha 1-->6)-2-deoxy-Ins, and GlcN(alpha 1-->6)Ins 1-dodecyl phosphonate inhibited GPI-PLC . GlcN(alpha 1-->6)Ins was as effective as Man-(alpha 1-->4)GlcN(alpha 1-->6)Ins; we surmise that GlcN(alpha 1-->6)Ins is the crucial glycan motif for GPI-PLC recognition . Inhibition by GlcN(alpha 1-->6)Ins 1,2-cyclic phosphate suggests product inhibition since GPIs cleaved by GPI-PLC possess a GlcN(alpha 1-->6)Ins 1,2-cyclic phosphate at the terminus of the residual glycan . The effectiveness of GlcN(alpha 1-->6)-2-deoxy-Ins indicates that the D-myo-inositol (Ins) 2-hydroxyl is not required for substrate recognition, although it is probably essential for catalysis . GlcN(alpha 1-->6)-2-deoxy-L-myo-inositol, unlike GlcN(alpha 1-->6)-2- deoxy-Ins, had no effect on GPI-PLC; hence, GPI-PLC can distinguish between the two enantiomers of Ins . Surprisingly, GlcN(alpha 1-->6)Ins 1,2-cyclic phosphate was not a potent inhibitor of Bacillus cereus PI-PLC, and GlcN(alpha 1-->6)Ins had no effect on the enzyme . However, both GlcN(alpha 1-->6)Ins 1-phosphate and GlcN(alpha 1-->6)Ins 1-dodecyl phosphonate were competitive inhibitors of PI-PLC . These observations suggest an important role for a phosphoryl group at the Ins 1-position in PI-PLC recognition of GPIs . Other studies indicate that abstraction of a proton from the Ins 2-hydroxyl is not an early event in PI-PLC cleavage of GPIs . Furthermore, both GlcN(alpha 1-->6)-2-deoxy-Ins 1-phosphate and GlcN(alpha 1-->6)-2-deoxy-L- myo-inositol inhibited PI-PLC without affecting GPI-PLC . Last, the aminoglycoside G418 stimulated PI-PLC, but had no effect on GPI-PLC . Thus, these enzymes represent mechanistic subclasses of GPI phospholipases C, distinguishable by their sensitivity to GlcN(alpha 1-->6)Ins derivatives and aminoglycosides . Possible allosteric regulation of PI-PLC by GlcN(alpha 1-->6)Ins analogs is discussed.

Biochem Biophys Res Commun, 1995 Feb 6, 207(1), 55 - 61
Terminal oxidases of the bb- and caa3-types in Bacillus sp . FTU; Muntyan MS et al.; We previously identified two oxidases in the membranes of bacterium Bacillus sp . FTU . One of them slowly (caa3) and the other rapidly (bo) recombines with carbon monoxide (CO) after laser flash photolysis, in this respect resembling the Escherichia coli bo- and bd-type oxidases, respectively . In the present study we found three copper atoms in the slowly CO-recombining oxidase from Bacillus sp . FTU . In the other oxidase, the copper content is very low and clearly substoichiometric . Reversed-phase chromatography revealed the presence of haems A and C in the Bacillus sp . FTU copper-containing oxidase and haems B and C in the non-copper-containing one . We thus suggest that the Bacillus sp . FTU oxidase rapidly reacting with CO previously attributed to bo-type by analogy in redox spectrum with the E . coli enzyme be redefined as bb-type oxidase.

Biochem Biophys Res Commun, 1995 Feb 6, 207(1), 20 - 4
Improvement of thermal stability of subtilisin J by changing the primary autolysis site; Bae KH et al.; The thermostability of subtilisin J, an extracellular serine protease secreted from Bacillus stearothermophilus, has been improved by changing the primary autolysis site of the Asp-49 mutant protein . Previously we have shown that the Asp-49 mutant protein has proteolytic activity, but so unstable that it was primarily autolyzed in Tyr-58-Gln-59 peptide bond during cultivation (Jang et al . Biochim . Biophys . Acta . 1162, 233-235 1993) . In the present study, to mitigate the autolytic degradation and increase the thermostability, we deleted the Tyr-58 residue using the Asp-49 mutant as a template . This mutant (Asp-49/delta Tyr-58 mutant) protein showed an improved resistance to heat treatment without changing the catalytic efficiency of the enzyme . These results show that change of primary autolysis site can stabilize the subtilisin.

Mol Gen Genet, 1995 Feb 6, 246(3), 301 - 8
Factors regulating cryIVB expression in the cyanobacterium--Synechococcus PCC 7942; Soltes-Rak E et al.; The expression of the larvicidal Bacillus thuringiensis subsp . israelensis cryIVB gene in cyanobacteria has been suggested to be an effective means of controlling mosquito populations . Using a variety of cryIVB constructs, in this study we have examined the effect of Synechococcus PCC 7942 culture age on intracellular toxin levels and have attempted to determine the mechanisms by which cryIVB gene expression is regulated . The data suggest that specific degradation of the cryIVB mRNA limits toxin production; however, the addition of cyanobacterial 3' untranslated DNA sequences to the cryIVB gene did not improve mRNA stability or toxin levels . An analysis of the cryIVB sequence and comparison of codon usage patterns with highly expressed cyanobacterial genes suggest that inefficient translation and intragenic ribosomal binding sites impede protein synthesis and result in rapid turnover of the toxin mRNA.

Hindustan Antibiot Bull, 1995 Feb-Nov, 37(1-4), 16 - 24
Production of free amino acids by some earthworm-borne microorganisms; Joy EK et al.; Production of free amino acids by some earthworm-borne microorganisms was investigated in three different synthetic media . Among the fungi tried Gliocladium roseum and Heterocephallum aurantiacum; among bacteria screened Bacillus macerans and B . mycoides; and among actinomycetes tested Streptomyces rimosus, S . violans, S . antibiticus, S . corchorusii and S . atroolivaceus produced significant amount of free amino acids . No correlations could be observed between vegetative growth and free amino acid production.

Hindustan Antibiot Bull, 1995 Feb-Nov, 37(1-4), 1 - 8
A carbon-limited medium for growth and sporulation of Bacillus thuringiensis var . kurstaki; Liu WM et al.; A culture medium for batch production of d-endotoxin by Bacillus thuringiensis (B., t.) has been modified . Through batch and continuous cultivation studies, the original medium was diagnosed to be limited in organic nitrogen . Corn steep liquor was found to be an excellent source for the organic nitrogen and its addition resulted in a carbon limited medium and in a significant increase in the amount of spore-toxin complex formed in shake flasks . Results of bioassay, conducted on Trichoplusia ni, suggest enhancement of larvicidal efficacy under carbon-limited growth conditions.

J Econ Entomol, 1995 Feb, 88(1), 97 - 105
Variation in tolerance to Bacillus thuringiensis among and within populations of the spruce budworm (Lepidoptera: Tortricidae) in Ontario; van Frankenhuyzen K et al.; Variation in tolerance to Bacillus thuringiensis Berliner subsp . kurstaki (strain HD-1-S-1980) among and within populations of the spruce budworm, Choristoneura fumiferana (Clemens), was assessed in the laboratory . Force-feeding assays using offspring of females collected as pupae from nine locations throughout Ontario and from a laboratory colony (DCF) demonstrated limited variation in tolerance among populations . Variation among populations was comparable with the variation observed among repeated assays with different batches of larvae from the DCF colony . Population LC50s were not significantly associated with age of the outbreak, host-plant species, incidence of the microsporidian Nosema fumiferanae (Thomson), or size of the female parent . Upper limits for genetic variation in tolerance were estimated by examining variation among full-sibling families within same populations . Mortality of individual families ranged from 6.5 to 70.9% within five field populations and from 2.7 to 93.3% within two laboratory colonies in response to a dose that caused a mean mortality of 40% . Familial factors accounted for 32.8% of the phenotypic variation in response across field populations, as compared with 3% for population factors . These data suggest that the phenotypic variation in tolerance to B . thuringiensis has a substantial genetic component and may provide a basis for evolution of resistance given sufficient selection pressure.

J Econ Entomol, 1995 Feb, 88(1), 21 - 6
Inheritance of resistance to Bacillus thuringiensis subsp . tenebrionis CryIIIA delta-endotoxin in Colorado potato beetle (Coleoptera: Chrysomelidae); Rahardja U et al.; We investigated the genetic inheritance of Colorado potato beetle, Leptinotarsa decemlineata (Say), resistance to Bacillus thuringiensis CryIIIA delta-endotoxin . Standard reciprocal crosses and backcrosses between susceptible (S) and resistant (R) strains were used to determine the characteristics of resistance . Analysis of probit lines from the F1 reciprocal crosses indicated that B . thuringiensis delta-endotoxin resistance was inherited autosomaly without maternal effects . We estimated the degree of dominance to be 0.77 and 0.76 for the (R x S) and (S x R) F1 generations, respectively, indicating that B . thuringiensis CryIIIA delta-endotoxin resistance is conferred by incompletely dominant genes . Chi-square analysis of mortality responses of backcrossed offspring suggested that resistance might be caused by more than one locus . The stability of resistance was also studied by testing seventeen generations of resistant beetles after the selection pressure was removed . When the selection pressure was removed, the resistance level of the selected colony decreased after five generations . The resistance level did not decrease further when the selection was removed for > 12 generations.

Cancer Immunol Immunother, 1995 Feb, 40(2), 103 - 8
The induction of bacillus-Calmette-Guérin-activated killer cells requires the presence of monocytes and T-helper type-1 cells; Thanhauser A et al.; Previously we have described the induction of MHC-unrestricted killer cells against bladder tumour cells by bacillus Calmette-Guerin (BCG), termed BCG-activated killer (BAK) cells . In the present paper we deal with the accessory-cell requirement for the activation of BAK cells . We show that monocytes are required for activating BAK cells, since no cytotoxicity can be induced in the absence of monocytes . Therefore, these phagocytes may represent the first step during the activation cascade of BAK cells . Furthermore, the presence of CD4+ T cells was essential for generating BAK cells: depleting peripheral blood mononuclear cells of CD4+ cells prior to stimulation with BCG abolished the cytotoxicity against bladder tumour cells . Experiments with monoclonal antibodies (mAb) neutralizing the activity of either interleukin-2 (IL-2) or interferon gamma (IFN gamma) underlined the importance of these cytokines: both mAb blocked the induction of BAK cells . Since both cytokines are related to the so-called Th1 pattern of T cells, we consider the second step of the generation of BAK cells as follows: monocytes presenting antigens of BCG trigger Th1-like cells in a preferred manner . These Th1-like T cells secrete IL-2 and IFN gamma and, thus, activate the BAK effector cells . Since CD4+ cells are dominant in the cells infiltrating the bladder wall after intravesical instillation of BCG in vivo, we postulate an important role for the Th1 subpopulation . We further postulate that the occurrence of macrophages in this infiltrate seems to be significant in the maintenance of the relapse-free state of the patient.

Eur J Immunol, 1995 Feb, 25(2), 377 - 84
Immune response to Mycobacterium bovis bacille Calmette Guérin infection in major histocompatibility complex class I- and II-deficient knock-out mice: contribution of CD4 and CD8 T cells to acquired resistance; Ladel CH et al.; Knock-out mice with defined major histocompatibility complex (MHC) deficiencies were infected intravenously with Mycobacterium bovis bacille Calmette Guerin (M . bovis BCG) to assess the relative impact of MHC class I- and II-dependent immune responses . Heterozygous control mice were capable of controlling growth of M . bovis BCG, although infection progressed chronically, as assessed by determination of colony-forming units . Furthermore, infected controls developed granulomatous lesions at the site of mycobacterial growth and delayed-type hypersensitivity (DTH) reactions after challenge with purified protein derivative of tuberculin . In vitro, spleen cells from heterozygous control mice produced high concentrations of interferon-gamma (IFN-gamma) after restimulation with mycobacterial antigens . In contrast, the MHC class II-deficient A beta-/- mice, which are virtually devoid of functional CD4 T cells, succumbed to M . bovis BCG infection . Furthermore, A beta-/- mice lacked DTH reactions to tuberculin and only few minute picnotic lesions were formed in livers of infected mice . Finally, spleen cells from infected A beta-/- mice failed to produce measurable IFN-gamma concentrations after restimulation in vitro with various mycobacterial antigen preparations . The capacity of beta 2-microglobulin (beta 2m)-deficient mice, which are devoid of CD8 alpha/beta T cells, to inhibit growth of M . bovis BCG was only slightly affected at low inocula, although significantly higher colony-forming units were detected in spleens . These knock-out mice developed strong DTH responses to tuberculin and their spleen cells produced high levels of IFN-gamma once reactivated by mycobacterial antigens . Furthermore, in livers of infected beta 2m-deficient mice, extravascular infiltrates developed which were more diffuse than those in infected control littermates . Remarkably, the beta 2m-deficient mice were substantially more susceptible to higher inocula of M . bovis BCG than their control littermates . Our data formally prove the essential role of MHC class II-dependent immune mechanisms in all relevant aspects of immunity to M . bovis BCG . In addition, our findings emphasize an important contribution of MHC class I-dependent immunity to effective anti-mycobacterial protection . We assume that CD4 T cells are highly effective in containing M . bovis BCG within distinct granulomatous lesions, but fail to eradicate their intracellular pathogens . It appears most likely that CD8 T cells are also required to achieve this goal.

Urol Clin North Am, 1995 Feb, 22(1), 139 - 50
A perspective on vesicoureteral reflux; Belman AB; Prevention of UTI appears to be the most important way to avoid the serious complications of vesicoureteral reflux, which then requires early recognition, ideally prior to bacterial invasion . With early evaluation of children noted to have dilated collecting systems in utero and the screening of siblings and offspring of those with reflux, this prevention becomes possible . This screening should be performed in the first weeks to months after birth, before the first UTI . The choice of management appears to be less important than control of infection, because the results of both medical and surgical management are equal; however, because mild-to-moderate (grades I-III) reflux is likely to resolve, it seems appropriate to pursue an aggressive nonsurgical course in these patients, at least until some minimally invasive, safe interventional treatment becomes available . If reflux remains severe (grades IV and V) beyond 24 to 48 months of age, surgical intervention appears appropriate because resolution is unlikely, assuming, of course, that an experienced surgeon performs the procedure . As was evident from the European branch of the IRS, renal scarring occurred most frequently in the few patients who had ureteral obstruction after failed surgical correction . In those who continued to have mild reflux beyond 5 to 7 years of age, a trial of medication is justifiable . If infection occurs during that time and reflux persists, correction should be considered for those with clinical or scan-documented pyelonephritis . Patients who have reflux plus bacteriuria present a special problem because it is unclear whether their risks are increased . Finally, we must forewarn all our female patients with UTI in childhood that they are at risk for bacilluria during pregnancy and may require prophylaxis regardless of the state of their reflux at that time.

J Infect Dis, 1995 Feb, 171(2), 459 - 62
Virulence of coccoid and bacillary forms of Helicobacter pylori in gnotobiotic piglets; Eaton KA et al.; This study sought to determine if coccoid forms of Helicobacter pylori are virulent for gnotobiotic piglets . Coccoid forms were generated by maintaining broth cultures of H . pylori under microaerobic conditions for 16 days . The resulting cultures contained bacteria with a coccoid morphology that could not be cultured in vitro . Coccoid H . pylori did not colonize any of 6 gnotobiotic piglets that were inoculated, whereas bacillary H . pylori colonized 6 of 6 inoculated piglets . Piglets colonized by bacillary H . pylori developed lymphocytic gastritis, but no gastritis developed in piglets inoculated with coccoid H . pylori, and coccoid-inoculated piglets were sero-negative for H . pylori-specific antibody . Thus, coccoid H . pylori appears to be a degenerate nonviable morphologic phase.

AJR Am J Roentgenol, 1995 Feb, 164(2), 283 - 6
Helicobacter pylori and peptic ulcer disease; Cello JP; Since William Beaumont's classic description of gastric physiology in 1847, the focus of clinical practice and basic research concerning gastric and duodenal ulcers has centered on the secretion of acid and pepsin . In addition to these well-known factors (largely determined by heredity, age, and oral intake), the search for other agents that alter the mucosal barrier has identified a bacillus, Helicobacter pylori, as a significant agent in the development of chronic gastritis and benign gastric and duodenal ulcers . This review explores the scientific evidence for an important causal role of H . pylori in the development of peptic ulcers on the basis of pathologic, pathophysiologic, and pharmacologic data.

J Bacteriol, 1995 Feb, 177(3), 745 - 9
Cloning and characterization of the gene for a protein thiol-disulfide oxidoreductase in Bacillus brevis; Ishihara T et al.; The gene (bdb) for protein thiol-disulfide oxidoreductase cloned from Bacillus brevis was found to encode a polypeptide consisting of 117 amino acid residues with a signal peptide of 27 residues . Bdb contains a well-conserved motif, Cys-X-X-Cys, which functions as the active center of disulfide oxidoreductases such as DsbA, protein disulfide isomerase, and thioredoxin . The deduced amino acid sequence showed significant homology with those of several bacterial thioredoxins . The bdb gene complemented the Escherichia coli dsbA mutation, restoring motility by means of flagellar and alkaline phosphatase activity . The Bdb protein overproduced in B . brevis was enzymatically active in both reduction and oxidization of disulfide bonds in vitro . Immunoblotting indicated that Bdb could function at the periphery of the cell.

Infect Immun, 1995 Feb, 63(2), 632 - 9
Extracellular virulence factors in Bacillus cereus endophthalmitis: methods and implication of involvement of hemolysin BL; Beecher DJ et al.; Bacillus cereus is a common cause of highly fulminant posttraumatic and metastatic endophthalmitis . Exotoxins or enzymes likely contribute to the severity of the infection, but specific virulence factors have not been identified . We developed two methods for the identification of B . cereus ocular virulence factors . In an in vitro assay that allows screening of multiple samples, retinal toxicity was estimated by measuring the release of lactate dehydrogenase from retinal buttons treated with B . cereus toxins . The results from this assay were confirmed with a sterile endophthalmitis model in which the histopathologic effect of intravitreally injected toxins was assessed . We tested pure hemolysin BL (HBL), a tripartite dermonecrotic vascular permeability factor of B . cereus, and crude exotoxin (CET) preparations, consisting of concentrated, cell-free B . cereus culture supernatant . In the in vitro assay, both CET and HBL caused rapid release of lactate dehydrogenase and retinal disintegration . In vivo, the toxins caused endophthalmitis clinically characteristic of B . cereus within 4 h . Histological changes included rapid retinal necrosis and detachment, choroidal edema, detachment and disruption of the retinal pigment epithelium, and rapid infiltration of polymorphonuclear leukocytes . Neutralization of HBL in CET preparations inhibited toxicity in vitro by 54%, and pure HBL was less toxic than CET with equal HBL contents in both methods . The results suggest that B . cereus ocular virulence is multifactorial and that HBL contributes to virulence.

Glycoconj J, 1995 Feb, 12(1), 99 - 107
Glycan structure of the S-layer glycoprotein of Bacillus sp . L420-91; Kosma P et al.; Preliminary taxonomic characterization of isolate L420-91 has revealed that this organism is closely related to the species Bacillus aneurinolyticus . The bacterium is covered by a squarely arranged crystalline surface layer composed of identical glycoprotein subunits with an apparent molecular mass in the range of 109 kDa . A total carbohydrate content of approximately 3.5% (wt/wt) was determined in the purified surface layer glycoprotein . Glycopeptides were obtained after exhaustive Pronase digestion and purification including gel filtration, ion exchange chromatography and HPLC . From the combined evidence of composition analysis . Smith degradation and nuclear magnetic resonance spectroscopy experiments we propose the following structure for the glycan chain of the surface layer glycoprotein: {formula: see text}

Biol Chem Hoppe Seyler, 1995 Feb, 376(2), 81 - 90
Determination of peptide regions exposed at the surface of the bacterial ribosome with antibodies against synthetic peptides; Herfurth E et al.; We synthesized six peptides corresponding to regions that are predicted to be surface-exposed of the following ribosomal proteins: protein L2, positions (D263-K272); protein L5, positions (I136-G150); protein L25, positions (Q75-D90); protein S3, positions (Q222-K232) derived from Escherichia coli; and protein L2, positions (K257-K275), and protein S3, positions (R130-T150) from Bacillus stearothermophilus . These peptides were employed to raise ribosomal protein-cross-reactive antibodies . The anti-peptide antisera reacted specifically with their parent proteins, as demonstrated by immunoblotting experiments . In a competition assay proteins L2 from E . coli and B . stearothermophilus as well as proteins L5 and L25 from E . coli were found to be accessible to the respective anti-peptide antibodies in the 50S subunits, but not in 70S ribosomes, proving their location at the 50S interface which is covered by the 30S subunit in the 70S complex . Two of the anti-peptide antisera directed against sequences deduced from protein S3 of E . coli and B . stearothermophilus reacted with 30S subunits as well as with 70S ribosomes, demonstrating their location at the backside, which is exposed to solvent . Thus, by the strategy applied specific short peptide stretches were located at the surface of the ribosome.

Mol Microbiol, 1995 Feb, 15(3), 403 - 10
Alkaliphiles: 'basic' molecular problems of pH tolerance and bioenergetics; Krulwich TA; Alkaliphilic Bacillus species provide experimental opportunities for examination of physiological processes under conditions in which the stress of the extreme environment brings issues of general biological importance into special focus . The alkaliphile, like many other cells, uses Na+/H+ antiporters in pH regulation, but its array of these porters, and other ion-flux pathways that energize and support their activity, result in an extraordinary capacity for pH homeostasis; this process nonetheless becomes the factor that limits growth at the upper edge of the pH range . Above pH 9.5, aerobic alkaliphiles maintain a cytoplasmic pH that is two or more units below the external pH . This chemiosmotically adverse delta pH is bypassed by use of an electrochemical gradient of Na+ rather than of protons to energize solute uptake and motility . By contrast, ATP synthesis occurs via completely proton-coupled oxidative phosphorylation that proceeds just as well, or better, at pH 10 and above as it does in the same bacteria growing at lower pH, without the adverse pH gradient . Various mechanisms that might explain this conundrum are described, and the current state of the evidence supporting them is summarized.

Int J Pept Protein Res, 1995 Feb, 45(2), 122 - 8
Mechanism of solvent-induced thermal stabilization of alpha-amylase from Bacillus amyloliquefaciens; Rajendran S et al.; The transition temperature of irreversible thermal inactivation of alpha-amylase from Bacillus amyloliquefaciens was estimated to be 60 degrees C . At this temperature, the enzyme inactivation followed first-order kinetics, having a half-life (t 1/2) of 12 min with a rate constant (k) of 0.06 min-1 . Conformational change was a prerequisite for this thermal inactivation . This is governed by stepwise temperature-dependent phenomena . Among the solvent stabilizers tested, the enzyme was thermally stable in presence of DMSO and PEG 300 and the stabilizing efficiency of these cosolvents was concentration-dependent . The enzyme was partially stabilized by 5.0 M DMSO and 1.9 M PEG 300 up to 78 degrees C . However, above 78 degrees C the enzyme was inactivated in these cosolvents also . The mechanism of stabilization has been explained by preferential hydration of the enzyme in these structure stabilizing solvents by exclusion from the protein surface and interface by measurement of partial specific volume in these cosolvents . The data suggest a high value of preferential interaction parameter, (delta g3/delta g2)tau, mu 1, mu 3 being -0.606/g/g g/g in 40% DMSO and a low value of -0.025 g/g in 5% glycerol . The preferential interaction parameters in sucrose and glycerol suggests that (delta g3/delta g2)tau, mu 1, mu 3m is highest of -0.420 g/g in 10% glycerol than any other cosolvent.

Biosci Biotechnol Biochem, 1995 Feb, 59(2), 236 - 9
Effect of galactooligosaccharides on calcium absorption and preventing bone loss in ovariectomized rats; Chonan O et al.; The effects of galactooligosaccharides (GOS), a mixture of galactosyl oligosaccharides formed from lactose by the transgalactosyl reaction of beta-D-galactosidase derived from Bacillus circulans, on calcium absorption and prevention of bone loss were examined in ovariectomized (OVX) Wistar rats . Rats fed on a diet containing GOS absorbed calcium more efficiently than those on the control diet after 8-10 days and 18-20 days, and the bone (femur and tibia) ash weight and tibia calcium content of OVX rats fed on the GOS diet were significantly higher than those of the control animals . Although the serum total cholesterol of the ovariectomized rats was significantly elevated, GOS produced a significant hypocholesterolemic effect in the OVX rats . GOS, which is fermented by bacteria in the lower part of the intestine, enhanced volatile fatty acid production, and thus prevented bone loss and lower serum total cholesterol concentration in the ovariectomized rats.

Biosci Biotechnol Biochem, 1995 Feb, 59(2), 231 - 5
Identification of DNA-binding proteins changed after induction of sporulation in Bacillus cereus; Matsuno K et al.; DNA-binding proteins were extracted from both exponentially growing cells of Bacillus cereus ts-4 and cells that were induced to sporulate at different stages of chromosome replication, by using a double-stranded B . cereus ts-4 DNA-cellulose column . Two-dimensional electrophoresis of the proteins found that the amounts of 17 proteins changed drastically after induction of sporulation at all the stages . For 8 of those proteins, the largest or the smallest amount was found in the cells which were induced to sporulate 40 min after the initiation of chromosome replication, the sensitive stage for sporulation . The N-terminal amino acids of 6 proteins among the selected proteins were sequenced . The sequence obtained from a 59-kDa protein had sequence similarity (> 45%) to GroEL from several bacterial species . In addition, the sequences from 76- and 52-kDa proteins matched deduced amino acid sequences of a Mycobacterium leprae gene showing homology to the bacteria atp operon and the B . subtilis guaB for IMP dehydrogenase, respectively.

Curr Microbiol, 1995 Feb, 30(2), 97 - 103
Bacteria produce the volatile hydrocarbon isoprene; Kuzma J et al.; Various bacterial species, both Gram-negative and Gram-positive, were found to produce the volatile hydrocarbon isoprene (2-methyl-1,3-butadiene) . Out of the tested cultures, Bacillus produced the most isoprene . The production of isoprene from bacteria was confirmed by gas chromatography-mass spectrometry . Media and growth effects on isoprene production were investigated: growth in rich media led to higher levels of isoprene than growth in minimal media, and highest isoprene emission rates were seen in log-phase cultures . Temperature profiles for bacterial isoprene production showed an optimum of 45 degrees C and were suggestive of an enzymatic mechanism for isoprene formation.

Compend Contin Educ Dent, 1995 Feb, 16(2), 136 - 8, 140, 142 passim; quiz 148
An overview of the oral manifestations of AIDS-related Kaposi's sarcoma; Flaitz CM et al.; Acquired immunodeficiency syndrome-associated Kaposi's sarcoma (AIDS-KS) is the most common malignancy in human immunodeficiency virus infection and is seen most often in homosexual men . The oral cavity is frequently involved by AIDS-KS and may represent the initial site of this malignancy in up to 60% of patients . A number of treatment modalities, including systemic and localized chemotherapy and radiotherapy, are available for AIDS-KS . The initial diagnosis of AIDS-KS requires microscopic evaluation of biopsy material because this disease can mimic a number of intraoral lesions, including atrophic candidiasis, erythroplakia, pyogenic granuloma, bacillary angiomatosis, median rhomboid glossitis, hemangioma, and lymphoma . Overall, treatment of AIDS-KS does not significantly affect the prognosis or survival of AIDS patients, however, treatment can alleviate aerodigestive and/or respiratory dysfunction, allow for adequate nutritional intake, and improve the quality of life for these patients.

Neuroscience, 1995 Feb, 64(4), 967 - 77
Demyelination in the central nervous system following a delayed-type hypersensitivity response to bacillus Calmette-Guérin; Matyszak MK et al.; We describe here a model of delayed-type hypersensitivity response in the CNS directed against a non-CNS antigen . The results presented in this paper show that bacillus Calmette-Guerin sequestrated behind the blood-brain barrier provokes an immune-mediated assault leading to bystander myelin damage . The delayed-type hypersensitivity response was induced by the intracranial injection of heat-killed bacillus Calmette-Guerin followed by subcutaneous immunization two to six weeks later . A single intracranial injection of bacillus Calmette-Guerin resulted in a rapid myelomonocytic response which persisted for approximately two weeks . By four weeks the inflammatory cells were no longer detected . Serum proteins were also excluded from the CNS parenchyma at this time . However, immunohistochemical staining with anti-bacillus Calmette-Guerin antiserum revealed the presence of bacillus Calmette-Guerin debris at the site of the original intracranial injection, indicating that the inflammatory response failed to clear the mycobacterium fully . Following peripheral sensitization with bacillus Calmette-Guerin in complete Freund's adjuvant, a strong delayed-type hypersensitivity response was detected at the site of bacillus Calmette-Guerin deposits in the CNS . An extensive inflammatory lesion was spread over a large area of the dorsal hippocampus . The lesion was composed predominantly of mononuclear phagocytes and T cells . Staining with anti-myelin basic protein antiserum showed bystander myelin damage . Delayed-type hypersensitivity responses were studied over several months and were still detected in the CNS five months after peripheral immunization.

J Neurol, 1995 Feb, 242(3), 147 - 52
Rapid diagnosis of tuberculous meningitis by polymerase chain reaction assay of cerebrospinal fluid; Lin JJ et al.; A polymerase chain reaction (PCR) method for the rapid diagnosis of tuberculous meningitis (TBM) was used to study prospectively 47 cerebrospinal fluid (CSF) samples from 45 patients . Twenty CSF samples were from patients with clinically suspected TBM and another 27 samples came from patients without clinically suspected TBM . Mycobacterial DNA was detected in 15 CSF samples (14 from patients with clinically suspected TBM and 1 from a patient not suspected of having TBM) . Of the PCR-positive samples, 4 were also positive for mycobacterial culture . However, 32 PCR-negative samples were all culture-negative . All samples were negative for the acid-fast bacillus by direct smear . The single PCR-positive patient in the clinically unsuspected TBM group was initially diagnosed as suffering from aseptic meningitis on the basis of his clinical features . The mycobacterial culture of his CSF specimen was also positive and a revised diagnosis of an aseptic type of TBM was made . The estimations of specificity and sensitivity in this study were 100% and 70% respectively . The results showed that using a PCR to detect mycobacterial DNA in CSF for the early diagnosis of TBM is not only a rapid but also an accurate method.

Immunology, 1995 Feb, 84(2), 317 - 21
Infection of Mycobacterium bovis bacillus Calmette-Guérin in antibody-mediated gamma delta T-cell-depleted mice; Nabeshima S et al.; To evaluate the hypothesis that gamma delta T cells participate in protective immunity against mycobacterial infection, we depleted gamma delta T cells from mice by administration of anti-T-cell receptor (TCR)gamma delta monoclonal antibody (mAb) and analysed protection against Mycobacterium bovis bacillus Calmette-Guerin (BCG) . The gamma delta T-cell-depleted mice did not show any exaggerated bacterial multiplication compared with control mice . In contrast, alpha beta T-cell-depleted mice, which were administrated anti-TCR alpha beta mAb before BCG infection, showed a depressed protective immunity . These results suggest that gamma delta T cells are not essential for coping with a primary BCG infection.

Biol Pharm Bull, 1995 Feb, 18(2), 350 - 4
Inactivation of blasticidin S by Bacillus cereus . V . Purification and characterization of blasticidin S-deaminase mediated by a plasmid from blasticidin S resistant Bacillus cereus K55-S1; Nawa K et al.; Blasticidin S (BS) deaminase (BSR) from a BS-resistant strain, Bacillus cereus K55-S1, was purified to homogeneity . Molecular weights determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and by gel filtration on HPLC are about 15500 and 35000, respectively, indicating the enzyme is a homodimer . The amino acid composition and N-terminal sequence of BSR are the same as those deduced from the nucleotide sequence of the BS-resistant gene, bsr . The optimum temperature and pH for enzyme activity are 60-65 degrees C and near 10.0, respectively . The activity of BSR is inhibited by Cu2+, Hg2+, and p-chloromercuric benzoate (PCMB) . Inhibition by PCMB or HgCl2 is reversible by the addition of SH reagents . The enzyme catalyzes the deamination of BS and its derivatives, but not cytosine nucleosides.

Tuber Lung Dis, 1995 Feb, 76(1), 59 - 64
Dysregulation of homeostasis of blood T-lymphocyte subpopulations persists in chronic multibacillary pulmonary tuberculosis patients refractory to treatment; Bose M et al.; DESIGN: The dysregulation of homeostasis of blood-T lymphocyte subpopulations was studied in 21 cases of chronic, multibacillary pulmonary tuberculosis refractory to treatment . The clinico-bacteriological and immunological parameters studied in these cases (Gr A) were compared with those of a group of 10 newly-diagnosed drug sensitive cases of pulmonary tuberculosis (Gr B) at the beginning of the study and after 3 months of chemotherapy for tuberculosis . The chronic cases were treated with drugs selected from a reserve line . 10 normal healthy individuals were included in this study as a control group . RESULTS: At the beginning of the study the mean CD4/CD8 lymphocyte ratios in the refractory cases (0.69) and the newly diagnosed cases (0.81) were significantly lower than those of the normal control subjects (1.84) . After 3 months of chemotherapy all but 3 of the newly-diagnosed cases showed clinical improvement, and all became sputum-negative . Their CD4/CD8 ratio recorded a rise to near normal (1.54) . On the contrary, following 3 months of reserve-line regimen, only 7 of the 21 group A cases showed sputum conversion . In all of the refractory cases, irrespective of sputum conversion, the CD4/CD8 ratio remained low (1.05) . CONCLUSION: This probably indicates that due to a long-standing bacillary load in drug resistant pulmonary tuberculosis patients the dysregulation of homeostasis of blood-T lymphocytes becomes persistent . This in turn delays their clinical and immunological recovery, even when therapy is adequate.

Tuber Lung Dis, 1995 Feb, 76(1), 28 - 34
The influence of BCG immunization on tuberculin reactivity in healthy Chilean women in the third trimester of pregnancy; Sepulveda RL et al.; SETTING: Tuberculin testing is an accepted method for screening pregnant women for tuberculosis . The interpretation of tuberculin reactivity in bacillus Calmette-Guerin (BCG)-immunized pregnant women is still in debate . OBJECTIVE: Four related issues were addressed: (1) The effect of pregnancy on tuberculin reactivity; (2) the effect of age differential on tuberculin reactivity; (3) the effect of repeated immunization with BCG; and (4) the risk of developing tuberculosis during pregnancy or a 3-year post-partum period . DESIGN: We performed tuberculin testing in 840 healthy Chilean women in the 32nd to 34th week of pregnancy; 807 had been immunized with 1 or more doses of BCG . There were 3 age groups: 177 were < or = 19 years old, 534 were 20-29 years old, and 129 were > or = 30 years old . All women in the study were followed at least 3 years post-partum . RESULTS: Women < or = 19 years old and non-pregnant women of similar age studied in the same geographical area had a similar distribution of the size of tuberculin reactions . Over 50% of all BCG-immunized women in each group had tuberculin reactions > or = 10 mm . A differential effect of different doses of BCG was significant only in 20- to 29-year-old women . None of the women in this study developed tuberculosis during pregnancy or a 3-year post-partum observation period . CONCLUSIONS: Healthy, BCG-immunized pregnant women may have positive tuberculin reactions without having an increased risk for tuberculosis . The incidence of tuberculosis and the BCG immunization status need to be considered in the development of policy recommendations for diagnosis and treatment of Mycobacterium tuberculosis infection in pregnant women.

Zhonghua Wai Ke Za Zhi, 1995 Feb, 33(2), 119 - 22
{Attachment of bacillus Calmette-Guerin to bladder cancer cells and the expression of interleukin-6 biological activity}; Zhong H et al.; In order to elucidate the mechanisms of antitumor efforts of bacillus Calmette-Guerin (BCG), the effects of attachment of BCG to bladder carcinoma cell line EJ and the role of fibronectin in the in vitro experiment were studied . At the same time, the IL-6 biological activity in the medium of EJ cells was determined and the IL-6 biological activity in presence of BCG compared . The results indicated that BCG attached strongly to EJ cells, and the maximal attachment to the cells with BCG occurred 48 hours after culture . Fibronectin could significantly increase the attachment and not acclerate the proliferation of EJ cells . In the presence of fibronectin-conditioned medium, the maximal percentage of the tumor cells with attached BCG was 74.22% . Under this condition of experiment, there was IL-6 biological activity in the medium of EJ cells . The treatment of EJ cells with BCG did not regulate the expression of IL-6.

J Biochem (Tokyo), 1995 Feb, 117(2), 276 - 82
Three conserved glycine residues in valine activation of gramicidin S synthetase 2 from Bacillus brevis; Saito M et al.; The translated product from the gene fragment containing the second and third domains of gramicidin S synthetase 2 was purified to an essentially homogeneous state . It showed valine- and ornithine-activating activity and the second domain was proved to be the valine-activating domain . Three mutant genes from Bacillus brevis Nagano, BI-3, E-4, and E-5 strains, which encode defective valine-activating domains of gramicidin S synthetase 2, were sequenced . By comparison with the wild-type gene, single point mutations of guanine to adenine were found at the three conserved glycine codons; the 5303rd guanine in BI-3, the 5378th guanine in E-4, and the 4967th guanine in E-5, which corresponded to codon changes of the 1768th glycine to glutamic acid and the 1793rd and the 1656th glycine to aspartic acid . Loss of valine-adenylation activity by mutation at the 1656th glycine proved the direct participation of the TSGT/STGXPKG motif in the adenylation reaction, and suggests that this glycine residue with the conserved lysine residue of the motif forms the phosphate-binding loop for ATP-binding . The 1793rd glycine is a member of the YGXTE motif which was also conserved among adenylate-forming enzymes except acetyl-CoA synthetases . The 1768th glycine residue appears to maintain the conformation of the active site for aminoacyl adenylation since this residue is retained among the adenylate-forming enzymes, though flanking regions are not conserved . These results suggest that these glycine residues are essential for adenylate formation in the antibiotic peptide synthetase family and some other adenylate-forming enzymes.

J Nutr Sci Vitaminol (Tokyo), 1995 Feb, 41(1), 51 - 60
Purification and properties of branched chain amino acid aminotransferase from gramicidin S-producing Bacillus brevis; Kanda M et al.; The branched chain amino acid aminotransferase {EC 2.6.1.42} was purified to a homogeneous state from a gramicidin S-producing strain of Bacillus brevis . The enzyme had a molecular weight of about 93,000 and consisted of two identical subunits, each with a molecular weight of about 47,000 . One pyridoxal phosphate is bound per subunit . In addition to branched chain amino acids, the enzyme uses L-phenylalanine and L-tryptophan as the amino donor, indicating that B . brevis branched chain amino acid aminotransferase has a broad substrate specificity for the amino donor . The enzyme utilized 2-oxoglutarate as the amino acceptor . The purified enzyme exhibits its absorption maxima at 332 and 427 nm at neutral pH.

Lab Anim Sci, 1995 Feb, 45(1), 22 - 6
Characterization of cilia-associated respiratory bacillus in rabbits and analysis of the 16S rRNA gene sequence; Cundiff DD et al.; The cilia-associated respiratory (CAR) bacillus is an unclassified, gram-negative bacterium that has been implicated as an etiologic agent of respiratory tract disease in laboratory rodents . A morphologically and antigenically similar organism has been identified in rabbits and is thought to be a related bacterium, although clinical signs of disease and histologic lesions are absent in infected rabbits . To compare the pathogenicity of rat- and rabbit-origin CAR bacillus isolates in rabbits, neonatal rabbits were experimentally infected with CAR bacillus isolates obtained from an infected rat and rabbit . Rabbits experimentally inoculated with rabbit-origin CAR bacillus had a nasal discharge, seroconverted and developed histologic lesions, whereas rabbits inoculated with rat-origin CAR bacillus seroconverted but did not have evidence of colonization of the respiratory tract . The CAR bacillus isolates were further examined at the genetic level by sequencing 1,261 base pairs of the 16S rRNA gene from six CAR bacillus isolates obtained from infected rabbits . A consensus sequence was obtained and compared with the analogous gene sequence data from rat-origin CAR bacillus isolates . Results indicated that these two organisms are distinctly different, with only 48.8% sequence homology . Comparison of the rabbit-origin 16S rRNA gene sequence with the database Genbank indicated that the organism is most closely related to members of the genus Helicobacter . Bacteria with the highest percentage of similarity with the rabbit-origin CAR bacillus were Helicobacter sp . strain Seymour and H . felis, with 91.1 and 90.8%, respectively . Findings of this study indicate that CAR bacillus isolates from rats and rabbits are host-specific and are different bacteria that belong to distinct genera.

Schweiz Rundsch Med Prax, 1995 Feb 1, 84(5), 122 - 6
{Cat scratch (?) disease}; Thurnheer U et al.; The case of a 24 year old female student who developed typical cat scratch disease following an insect sting invited to review the literature . Over the last two years, Rocalimaea henselae could be identified and is now considered to be responsible for the majority of infected cases . It is related to Rochalimaea quintana that provokes trench fever and to Bartonella bacilliformis, known as the inciting agent of the verruca peruana and of Oroya fever . Recently, it has been proposed to integrate Rochalimaea and Bartonella into the family of Bartonellaceae . The diverse and fascinating array of clinical syndromes caused by R . henselae as well as their treatment are described . Capability to affect organs other than skin and lymph nodes is primarily but not exclusively associated with immunodeficiencies . In countries like Switzerland little information on these clinical pictures is available . However, it is important to recognize the typical symptoms and signs in order to initiate the appropriate and necessary examinations.

Eur J Biochem, 1995 Feb 1, 227(3), 687 - 93
Substrate specificity and detailed characterization of a bifunctional amylase-pullulanase enzyme from Bacillus circulans F-2 having two different active sites on one polypeptide; Kim CH et al.; Bacillus circulans F-2 amylase-pullulanase enzyme (APE) displayed dual activity with respect to glycosidic bond cleavage . The enzyme was active on alpha-1,6 bonds in pullulan, amylopectin, and glycogen, while it showed alpha-1,4 activity against malto-oligosaccharides, amylose, amylopectin, and soluble starch, but not pullulan . Kinetic analysis of the purified enzyme in a system which contained both pullulan and amylose as two competing substrates was used to distinguish the dual specificity of the enzyme from the single-substrate specificity known for pullulanases and alpha-amylases . Enzyme activities were inhibited by some metal ions, and by metal-chelating agents with a different mode . The enzyme-inhibitory results of amylase and pullulanase with Hg2+ and Co2+ ions were different, indicating that the activation mechanisms of both enzyme activities are different . Cyclomaltoheptaose inhibited both alpha-amylase and pullulanase activities with inhibition constants (Ki) of 0.029 and 0.06 mg/ml, respectively . Modification with 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide confirmed a carboxy group at the active sites of both enzymes . The N-terminal sequence of the enzyme was: Ala-Asp-Ala-Lys-Lys-Thr-Pro- Gln-Gln-Gln-Phe- Asp-Ala-Leu-Trp-Ala-Ala-Gly-Ile-Val-Thr-Gly-Thr-Pro-Asp-Gly-Phe . The purified enzyme displayed Michaelis constant (Km) values of 0.55 mg/ml for amylose, and 0.71 mg/ml for pullulan . When both amylose and pullulan were simultaneously present, the observed rate of product formation closely fitted a kinetic model in which the two substrates are hydrolyzed at different active sites . These results suggest that amylopullulanases, which possess both alpha-1,6 and alpha-1,4 cleavage activities at the same active site, should be distinguished from APEs, which contain both activities at different active sites on the same polypeptide . Also, it is proposed that the Enzyme Commission use the term 'amylase-pullulanase enzyme' to refer to enzymes which act on starch and cleave both alpha-1,6-bonds in pullulan and alpha-1,4 bonds in amylose at different active sites.

Med Clin (Barc), 1995 Jan 28, 104(3), 81 - 4
{Tuberculosis in La Rioja from 1988 to 1993 . Clinical course, localization, influence of human immunodeficiency virus infection, and sensitivity of mycobacterial tests}; Labarga Echeverria P et al.; BACKGROUND: The present study was performed to analyze the evolution of the incidence of tuberculosis in La Rioja (Spain) from January 1988 to March 1993 . The influence of factors such as HIV infection and alcoholism was analyzed . The sensitivity of the bacilloscopy in the diagnosis was studied . METHODS: The cases were collected from the Regional Tuberculosis Program of the Public Health Department in La Rioja, with clinical histories being collected from the Internal Medicine Department of the Hospital San Millan in Logrono . Cases of tuberculosis were microbiologically confirmed by cultures of Mycobacterium tuberculosis . RESULTS: The total number of cases studied was 649 with the incidence of tuberculosis in La Rioja being 51 cases/100,000 inhabitants in 1992 . The mean age of all the patients was 45 years with 80% of the tuberculosis being pulmonary and 20% extrapulmonary . Twelve percent of the patients were HIV+; 18% being extrapulmonary tuberculosis and 10.8% pulmonary tuberculosis, p < 0.02 . Pleural and urogenital tuberculosis were present in fewer HIV+ patients . Bacilloscopy was negative requiring a culture for diagnosis in 28.6% of the cases with this circumstance more frequently occurring in the pleural and urogenital forms of the diseases . Treatment has evolved from the use of ethambutol in the first few years to the present use of pyrazinamide . CONCLUSIONS: No trend towards an increase in the incidence of tuberculosis in the time period studied was observed in La Rioja . Extrapulmonary tuberculosis was the type of tuberculosis most frequently found in HIV positive patients, with pleural and urogenital forms of the disease being less frequently found in these patients . In these case the bacilloscopy was also found to be negative more frequently.

J Biol Chem, 1995 Jan 27, 270(4), 1489 - 92
Construction and function of chimeric beta subunits containing regions from the beta subunits of the F1F0 ATPases of Escherichia coli and Bacillus megaterium; Matten SR et al.; The highly conserved beta subunit of the Escherichia coli F1F0 ATPase was divided into three sections, each of which was exchanged with the homologous section of the beta subunit of the obligate aerobe Bacillus megaterium . Plasmids coding for the resultant six chimeric beta subunits varied in their abilities to complement two E . coli beta mutants as measured by testing transformed cells for aerobic growth on a nonfermentable carbon source or anaerobic growth on rich medium containing glucose . Two chimeras were able to restore both growth on succinate and anaerobic growth on rich medium . The genetic results corresponded to increased levels of membrane-bound ATPase and ATP synthase activities . These chimeric subunits were therefore capable of being assembled into functional E . coli ATPase complexes . The results indicate that chimeric beta subunits can be used to analyze assembly of the beta subunit and that the final 181 amino acids of the beta subunit might contain a region involved in functional energy coupling.

Presse Med, 1995 Jan 21, 24(3), 183 - 8
{Cat-scratch disease and disease caused by Bartonella (Rochalimaea)}; Drancourt M et al.; The aetiology of cat scratch disease remains controversial since both Afipia felis and Bartonella (Rochalimaea) henselae have been isolated from diseased lymph nodes . Bartonella henselae, Bartonella (Rochalimaea) quintana and Bartonella (Rochalimaea) elizabethae cause endocarditis and Bartonella bacilliformis cause septicemia (Oroya's fever) in non-immunocompromized patients, and Bartonella henselae and Bartonella quintana cause fever, bacillary angiomatosis, and visceral peliosis in human immunodeficiency virus-infected patients . Bartonella quintana is the historical agent of trench fever and we recently isolated it from chronic adenopathy . The diagnosis of Afipia felis and Bartonella infections relies upon the isolation of the bacterium from blood, node tissue after inoculation of cell cultures systems and molecular identification, and upon the serology . In vitro both species are sensitive to aminoglycosides, and we recommend aminoglycosides be included in antibiotic regimens for treating cat scratch disease and Bartonella infections.

Biochem Biophys Res Commun, 1995 Jan 17, 206(2), 577 - 83
A bifunctional monocyclic beta-lactam cross-links across the active site of beta-lactamase; Ahluwalia R et al.; A 4-alkoxy-2-azetidinone behaves as a bifunctional active site-directed inhibitor of the class A beta-lactamase from Bacillus cereus 569/H . It cross-links SER 70 and LYS 234 as it binds in a approximately 1:1 ratio . The cross-linked enzyme is irreversibly inhibited while the secondary structure is partially stabilized under conditions when the native enzyme is otherwise converted to a form with no detectable secondary structure by circular dichroism.

FEMS Microbiol Lett, 1995 Jan 15, 125(2-3), 287 - 92
Cloning and sequencing of the pac gene encoding the penicillin G acylase of Bacillus megaterium ATCC 14945; Martin L et al.; The pac gene encoding the penicillin G acylase (PGA) of Bacillus megaterium ATCC 14945 has been cloned in Escherichia coli HB101 (proA, leuB) using a selective minimal medium containing phenylacetyl-L-leucine instead of L-leucine . The nucleotide sequence of this gene has been determined and contains an open reading frame of 2406 nucleotides . The deduced amino acid sequence shows significant similarity with other beta-lactam acylases . Although the PGA of B . megaterium is extracellular, the enzyme produced in E . coli appears to have a cytoplasmic localization.

Proc Natl Acad Sci U S A, 1995 Jan 3, 92(1), 195 - 9
FALL-39, a putative human peptide antibiotic, is cysteine-free and expressed in bone marrow and testis; Agerberth B et al.; PR-39, a proline/arginine-rich peptide antibiotic, has been purified from pig intestine and later shown to originate in the bone marrow . Intending to isolate a clone for a human counterpart to PR-39, we synthesized a PCR probe derived from the PR-39 gene . However, when this probe was used to screen a human bone marrow cDNA library, eight clones were obtained with information for another putative human peptide antibiotic, designated FALL-39 after the first four residues . FALL-39 is a 39-residue peptide lacking cysteine and tryptophan . All human peptide antibiotics previously isolated (or predicted) belong to the defensin family and contain three disulfide bridges . The clone for prepro-FALL-39 encodes a cathelin-like precursor protein with 170 amino acid residues . We have postulated a dibasic processing site for the mature FALL-39 and chemically synthesized the putative peptide . In basal medium E, synthetic FALL-39 was highly active against Escherichia coli and Bacillus megaterium . Residues 13-34 in FALL-39 can be predicted to form a perfect amphiphatic helix, and CD spectra showed that medium E induced 30% helix formation in FALL-39 . RNA blot analyses disclosed that the gene for FALL-39 is expressed mainly in human bone marrow and testis.

Acta Microbiol Pol, 1995, 44(3-4), 315 - 316
Identification of a new restriction endonuclease R . BcrAI, from Bacillus cremoris; Piekarowicz A et al.; Site specific restriction endonuclease R . BcrAI has been purified from Bacillus cremoris . The enzyme recognize the sequence 5' CTCTTC 3'.

Emerg Infect Dis, 1995 Jan-Mar, 1(1), 16 - 21
Unraveling mysteries associated with cat-scratch disease, bacillary angiomatosis, and related syndromes; Regnery R et al.; The search for the infectious agents responsible for cat-scratch disease, bacillary angiomatosis, and related syndromes has a long and often circuitous history . Recognition of the etiologic agents and a new understanding of the fundamental features of the epidemiology and natural history of modern day Bartonella (formerly Rochalimaea)-associated diseases culminate a multipartite story that combines clinical medicine, traditional microbiology, and novel technological approaches to solve a long-standing enigma.

Nucleic Acids Symp Ser, 1995, (34), 99 - 100
Structure-function relationship of the xylanase from alkaliphilic Bacillus sp . strain 41M-1; Nakamura S et al.; Xylanase J from alkaliphilic Bacillus sp . strain 41M-1 contains a family G catalytic domain at the N-terminus, followed by a linker sequence and a functionally-unknown C-terminal domain . The mutational analysis of xylanase J indicated that Glu-93, Glu-183, Trp-18, Trp-86, Tyr-84 and Tyr-95 play an important role in the catalytic activity . A deletion derivative of xylanase J lacking the C-terminal domain retained its activity, suggesting that the C-terminal domain does not directly involved in catalysis.

Folia Microbiol (Praha), 1995, 40(4), 413 - 6
Prolonged survival of AVN Wistar rats with transplanted Yoshida sarcoma and increase of granular lymphocytes after administration of Bacillus firmus and their crude lipids; Stepankova R et al.; Bacillus firmus is a Gram-positive, aerobic, sporulating, nonpathogenic air contaminant which, according to earlier findings, is a strong polyclonal activator of B lymphocytes . The crude lipids of this microbe induced significant resistance of mice against listerial infection . The administration of bacterin, like that of crude lipids obtained by the extraction of cell suspension with chloroform-methanol to rats, strain AVN Wistar, transplanted later with Yoshida sarcoma, significantly prolonged the survival of the animals in comparison with the control group . At the same time the number of granular lymphocytes was increased . The destruction of tumor cells in the peritoneal exudate of immunostimulated rats was also determined.

Int Urol Nephrol, 1995, 27(6), 723 - 33
Evaluation of a low-dose intravesical bacillus Calmette-Guérin (Tokyo strain) therapy for superficial bladder cancer; Takashi M et al.; To examine whether intravesical instillation of low-dose bacillus Calmette-Guerin (BCG) therapy is effective with low toxicity, we reviewed data for 111 patients with superficial bladder cancer (stages Ta and T1) . Among them, 74 received the BCG treatment for prophylaxis of intravesical recurrence after transurethral resection, and the remaining 37 therapeutically for Ta or T1 tumours . The patients were divided into two groups by instillation dose of BCG (Tokyo 172 strain): 40 mg (n = 55) and 80 mg (n = 56), and statistically compared for recurrence, antitumour effect and toxicity . The mean instillations were done 8.4 times in the 40 mg dose group and 8.6 times in the 80 mg dose group . Among the 74 patients with BCG therapy for prophylaxis those with a previous episode of bladder cancer treatment (n = 47) experienced a significantly (p = 0.006) shorter recurrence-free interval than those with no episode (n = 27) . Among 47 patients with a previous treatment episode, those receiving the 80 mg dose demonstrated a significantly longer recurrence-free interval than those given the 40 mg dose (p = 0.03) . Among the 27 patients without previous treatment no significant difference in recurrence-free intervals was found between the two dose groups . Univariate and multivariate analyses using Cox's proportional hazards model confirmed the above findings . The recurrence index was also significantly reduced after BCG therapy in the 80 mg and 40 mg groups and similar antitumour effects for Ta or T1 tumours were observed in the two dose groups . The degree of toxicity due to BCG therapy was significantly (p = 0.02) lower with the 40 mg dose . The present study suggests that (1) a 40 mg BCG low-dose (Tokyo 172 strain) regimen is useful for preventing recurrence, with sufficient therapeutic efficacy and low frequency of toxicity, among patients without a previous treatment, and (2) prophylactic effects with the 80 mg dose regimen are much superior for previous treatment cases.

Ciba Found Symp, 1995, 195, 123 - 32; discussion 132-6
T cells and cytokines in intracellular bacterial infections: experiences with Mycobacterium bovis BCG; Kaufmann SH et al.; Intracellular bacteria reside in mononuclear phagocytes, and protective immunity is dominated by T lymphocytes . Mycobacterium bovis bacillus Calmette-Guein (BCG) infection of mice represents an excellent model for studying immune mechanisms involved in defence against persistent intracellular bacteria that cause chronic disease . Gene disruption mutant mice include: A beta-/-, which lack conventional CD4+ T cell receptor alpha/beta (TCR alpha/beta) T lymphocytes; beta 2 microglobulin -/-, which lack conventional CD8+ TCR alpha/beta lymphocytes; TCR beta-/-, which lack all TCR alpha/beta lymphocytes; TCR delta-/-, which lack all TCR gamma/delta lymphocytes; and RAG-1-/- mutants, which lack mature T and B lymphocytes . Studies of these mutants suggest that CD4+ TCR alpha/beta, CD8+ TCR alpha/beta and TCR gamma/delta T lymphocytes all contribute to immunity against M . bovis BCG . Activation of antibacterial effector functions in macrophages by T helper 1 (Th1) cell-derived gamma-interferon (IFN-gamma) is central to protection . In contrast, Th2 cells are only marginally involved . Activation of Th1 and Th2 cells is regulated by interleukin 10 (IL-10) and IL-12, which are induced early in infection with M . bovis BCG . Although IL-12 is stimulated by M . bovis BCG in immunocompetent mice, studies with IFN-gamma receptor-deficient and tumour necrosis factor alpha (TNF-alpha) receptor-deficient mutant mice suggest that M . bovis BCG-induced IL-12 secretion depends on IFN-gamma and TNF-alpha . Hence, IL-12 cannot be the first cytokine produced during M . bovis BCG infection.

Pneumoftiziologia, 1995 Jan-Jun, 44(1-2), 29 - 32
{The role of the epidemiological survey in childhood tuberculosis}; Mihailescu P et al.; All epidemiological inquiries in tuberculous children are centralized and processed in the Institute of Pneumophtisiology . The analysis of 4,171 inquiries processed during 1985-1992 shows: after 1985 an increase of diseased children is noted . This increase is higher in gipsy children, whose weight reached 38.7% out of the total in 1992; the tb . children were found out mainly by symptoms and in 20-25% by ascendent epidemiological inquiry; the bacillary sources which infected the children have been found out by ascendent epidemiological inquiry in 70% of cases; the infection sources consisted mainly in new lung tb . cases in adults (78-82%), relapses (6-8%) and chronic cases (12-14%) . In certain areas, the weight of chronic cases as infection sources exceded 20%.

Yi Chuan Xue Bao, 1995, 22(4), 322 - 8
{Molecular cloning of cellulase gene from the bacillus}; Guan J et al.; The CMCase gene from the Bacillus sp . strain E2 was cloned into E . coli DH5 alpha F' using plasmid pBR322 as a vector . Recombinant plasmid pBG3 was isolated from transformants producing CMCase, and the cloned CMCase gene was found to be in a 4.0kb HindIII fragment . The CMCase gene was subcloned in a functional state on a 2.4kb DNA fragment . The optimum pH and temperature for activity of CMCase encoded by pBG3 were pH 6.5 and 55 degrees C respectively . The CMCase could not hydrolyze any natural cellulose examined . Eighty-three percent of the CMCase was detected in the inner cellular fraction, 9% and 8% were in the periplasmic space and medium respectively . Result of Southern hybridization indicates that the cloned DNA fragment was derived from Bacillus sp . strain E2 chromosomal DNA.

Rev Mal Respir, 1995, 12(6), 593 - 9
{Palliative percutaneous treatment under x-ray computed tomographic control of inoperable pulmonary aspergilloma . Apropos of 30 cases}; Giron J et al.; The authors report 30 cases of the percutaneous treatment of symptomatic pulmonary aspergilloma by injection of amphotericine paste in patients who were not considered to be operable . The treated aspergillomas had developed as a sequel to bacilliary infection and pulmonary fibrosis . Surgery was contraindicated in these patients on account of severe respiratory failure . The authors specify the technique for the preparation of the paste and for the type of percutaneous injection, the aim being to obtain complete filling of the cavity and creating an anaerobic environment for the aspergillus . The contribution of this technique for the non-surgical treatment of patients appears interesting but should be carried on a larger series to identify the exact indications and the interaction with other new treatments which have just appeared.

Nucleic Acids Symp Ser, 1995, (33), 5 - 7
Protein recognition of a ribosomal RNA tertiary structure; Draper DE et al.; Ribosomal protein L11 recognizes a highly conserved, 58 nucleotide domain of large subunit ribosomal RNA . This domain has a set of tertiary interactions that are specifically stabilized by Mg2+ and NH4+ ions . The protein recognizes this tertiary structure, in the sense that ions stabilizing the tertiary structure also promote L11 binding, and a heat stable form of the protein (from Bacillus stearothermophilus) prevents RNA unfolding . We also find that these RNA-binding properties are confined to a approximately 75 amino acid domain of the protein . The larger issue of L11 function within the ribosome is discussed in light of these findings.

Arch Inst Pasteur Madagascar, 1995, 62(1), 99 - 102
{Peritoneal tuberculosis in Madagascar, 55 cases at the Soavinandriana Hospital Center in Antananaviro}; Peghini M et al.; This retrospective study conducted in Antananarivo for a 42 months period, from September 1991 to March 1994, allowed to record 55 peritoneal tuberculosis, all of them in malagasy patients . The sex ratio was 0,83 and the mean age 36 years old . Ascites puncture was done each time there was an effusion (44 cases) . For all cases . For all cases, the diagnosis based on laparoscopy allowed an investigation of the liver and the peritoneum, and 10 peritoneum biopsies could be done . The tubercular bacillus has been isolated in 2 ascites fluids out of 8 incubations, and in 5 biopsies of peritoneal granulation out of 8 . For 8 cases, another tubercular localization has been discovered: 5 pleurisis, 2 evolutive tuberculosis and 1 pericardial effusion . All the patients have been put under antitubercular treatment and have regained health . Peritoneal tuberculosis has become rare in the West but is still a frequent pathology mostly in young women, in Africa and up in Madagascar.

Arch Inst Pasteur Madagascar, 1995, 62(1), 52 - 8
{Evolution of the Mycobacteria Laboratory of the Pasteur Institute of Madagascar from 1991 to 1994}; Menard D et al.; In 1991, the Laboratory of Mycobacteria was a small laboratory, part of the Clinical Biology Centre (CBC) of the Institut Pasteur de Madagascar: 656 pathological samples have been analysed for the account of the CBC and the National Control Programme activities . Within 4 years, the number of samples tested increased by more than threefold and the technical ability has evolved in an important way, specially for the identification and the antibiotic sensitivity testing . The scientific equipment have been modernized and the rooms surface increased by fourfold . In 1995, this laboratory was officially designated as the National Reference Laboratory for the culture, the identification and antibiogramme for the account of the National Control programme and for the private clinicians . It also participates to the tuberculosis research programmes of Institut Pasteur de Madagascar . It is associated to the Laboratory of Mycobacteria in the Institut d'Hygiene Sociale of Antananarivo which is the National Reference Laboratory for the bacilloscopy, the teaching and the supervision of the peripheral laboratories.

Biopolymers, 1995, 37(6), 411 - 9
The suitability of a monofunctional reagent of an undecagold cluster for phasing data collected from the large ribosomal subunits from Bacillus stearothermophilus; Bartels H et al.; An electron density map of the large ribosomal subunit from Bacillus stearothermophilus was obtained at 26 A resolution by single isomorphous replacement (SIR) from a derivative formed by specific quantitative labeling with a dense undecagold cluster . For derivatization, a monofunctional reagent of this cluster was bound to a sulfhydryl group of a purified ribosomal protein, which was in turn reconstituted with core particles of a mutant lacking this protein . The native, mutated, and derivatized 50S ribosomal subunits crystallize under the same conditions in the same space group . Under favorable conditions, crystals of the derivatized subunit proved to be isomorphous with the native ones, whereas the crystals of the mutant may have somewhat different packing . After resolving the SIR phase ambiguity by solvent flattening, the electron density shows a packing that is consistent with the noncrystallographic symmetry found by Patterson searches as well as with the motif observed in electron micrographs of thin sections of the crystals . These studies established that phase information can be obtained from heavy metal clusters, even when the crystals under investigation are unstable and weakly diffracting . These results encouraged further effort at the construction of specifically derivatized crystals from other ribosomal particles that diffract to higher resolution.

Biochimie, 1995, 77(4), 256 - 61
Expression and secretion of malarial parasite beta-tubulin in Bacillus brevis; Bell A et al.; Microtubule inhibitors are active against the human malarial parasite Plasmodium falciparum, but whether these drugs actually interact with parasite tubulins is not known . It has not previously been possible to produce mg quantities of isolated, soluble tubulin subunits for drug-binding experiments . A cDNA encoding P falciparum beta-tubulin was expressed and the protein secreted in Bacillus brevis . With the addition of EGTA to the culture medium, which increases shedding of proteins from the cell surface, up to 2 mg/l recombinant beta-tubulin was obtained in supernatants . It is not clear why B brevis is able to secrete this normally cytoplasmic protein, but the secretion levels of recombinant proteins may be related to the net charge of the first few residues of the mature polypeptide.

Crit Rev Oncog, 1995, 6(1), 47 - 56
Verruga peruana: an infectious endemic angiomatosis; Caceres-Rios H et al.; Microbial-related dysplastic and neoplastic angiomatous proliferative processes are seen with increased frequency, particularly in the acquired immunodeficiency syndrome (AIDS) . The microbial-encoded or -induced mediators of angiopathogenesis in AIDS-associated Kaposi's sarcoma and bacillary angiomatosis are actively being sought . The present review addresses the historical, epidemiologic, clinical, etio- and histopathogenic aspects of the verruga peruana (VP) . VP is a disease thus far endemic to high Andean valleys and characterized by dermal angioblastic proliferation in association with reactivation of latent Bartonella bacilliformis organisms . VP closely resembles AIDS-associated angiopathogenic manifestations at the clinical, histopathologic, and etiologic levels and therefore has been proposed as a model for the study of angiogenesis and endothelial cell dysplasia and neoplasia . Moreover, the recent epidemic outbreaks in endemic areas, the increased frequency of international travel to the region, the variable incubation period, and the possibility of not recognizing VP due to its rarity further underscore the relevance of studying this rare disorder and of including it in the differential diagnosis of angiomatous-proliferative disorders.

Biomed Pharmacother, 1995, 49(7-8), 369 - 74
Triggering effects of BCG vaccine on antitumor and interleukin-1 secretory activity of T cell lymphokine-primed macrophages; Seledtsova GV et al.; By using an in vitro model, this work demonstrates that BCG (Bacillus Calmette-Guerin) vaccine is able to trigger activation of T cell lymphokine-primed murine macrophages (Mo) for both tumoricidal cytotoxicity and interleukin-1 (IL-1) secretion . The results presented support the view that BCG vaccine may essentially activate primed, but not intact Mo; and that macrophage-mediated both antitumor and immunostimulatory effects of BCG vaccine observed in the body may be largely defined by functional activity of T-lymphocytes responsible for antigen-induced production of macrophage-activating lymphokines.

Vet Res Commun, 1995, 19(5), 425 - 32
Some pharmacokinetic parameters of doxycycline in east African goats after intramuscular administration of a long-acting formulation; Ole-Mapenay IM et al.; A compartmental and non-compartmental study was carried out on five adult goats following intramuscular administration of doxycycline at 20 mg/kg bodyweight . The concentration of the drug in serum was determined by a microbiological assay employing Bacillus cereus var mycoides (ATCC 11778) as the test organism . The mean serum concentration (Cmax) and the time of maximum concentration (Tmax) were 1.87 micrograms/ml and 0.85 h, respectively . Using compartmental analysis, the plasma concentration-time curve of doxycycline best fitted a three-compartment open model with first-order absorption . A three-phase disposition of doxycycline was found, the terminal elimination half-life being approximately 40 h . The statistical moment theory was mainly used for non-compartmental analysis . The value obtained for the mean residence time (MRT) was 16.4 h . The mean values for the volume of distribution at steady state (Vdss), determined by compartmental and non-compartmental analyses, were 8.73 and 13.19 L/kg, respectively . There were no statistically significant differences when the major pharmacokinetic parameters were compared . It was concluded that the pharmacokinetic behaviour of doxycycline in goats after intramuscular administration is characterized by a three-compartment model with a slow terminal elimination phase . Based on current knowledge, this could be due to enterohepatic recycling and/or flip-flop kinetics . The study indicated that a single intramuscular administration of 20 mg/kg of doxycycline may only provide therapeutic concentrations for up to 24 h owing to slow absorption at the injection site.

Mem Inst Oswaldo Cruz, 1995 Jan-Feb, 90(1), 97 - 107
Transfer of toxin genes to alternate bacterial hosts for mosquito control; Orduz S et al.; Mosquitoes are vector of serious human and animal diseases, such as malaria, dengue, yellow fever, among others . The use of biological control agents has provide an environmentally safe and highly specific alternative to the use of chemical insecticides in the control of vector borne diseases . Bacillus thuringiensis and B . sphaericus produce toxic proteins to mosquito larvae . Great progress has been made on the biochemical and molecular characterization of such proteins and the genes encoding them . Nevertheless, the low residuality of these biological insecticides is one of the major drawbacks . This article present some interesting aspects of the mosquito larvae feeding habits and review the attempts that have been made to genetically engineer microorganisms that while are used by mosquito larvae as a food source should express the Bacillus toxin genes in order to improve the residuality and stability in the mosquito breeding ponds.

Mem Inst Oswaldo Cruz, 1995 Jan-Feb, 90(1), 81 - 6
Biochemistry and mode of action of the Bacillus sphaericus toxins; Davidson EW; Bacillus sphaericus produces at least two toxins which are highly toxic to mosquito larvae . The binary toxin, which is comprised of proteins of 51.4 and 41.9 kDa, is present in all highly insecticidal strains . The 100 kDa SSII-1 toxin is present in most highly insecticidal as well as the weakly insecticidal strains . The current status of studies on biochemistry and mode of action of these toxins is reviewed.

Mem Inst Oswaldo Cruz, 1995 Jan-Feb, 90(1), 75 - 9
Exploration of receptor binding of Bacillus thuringiensis toxins; Kwak IS et al.; Wild type and mutant toxins of Bacillus thuringiensis delta-endotoxins were examined for their binding to midgut brush border membrane vesicles (BBMV) . CryIAa, CryIAb, and CryIAc were examined for their binding to Gypsy moth (Lymantria dispar) BBMV . The binding of CryIAa and CryIAc was directly correlated with their toxicity, while CryIAb was observed to have lower binding than expected from its toxicity . The latter observation confirms the observation of Wolfersberger (1990) . The "rule" of reciprocity of binding and toxicity is apparently obeyed by CryIAa and CryIAc, but broken by CryIAb on L . dispar . Alanine substitutions were made in several positions of the putative loops of CryIAa to test the hypothesis that the loops are intimately involved in binding to the receptor . The mutant toxins showed minor shifts in heterologous binding to Bombyx mori BBMV, but not enough to conclude that the residues chosen play critical roles in receptor binding.

Mem Inst Oswaldo Cruz, 1995 Jan-Feb, 90(1), 69 - 74
Mechanism of action of Bacillus thuringiensis toxins; Gill SS; The selectivity of Bacillus thuringiensis toxins is determined both by the toxin structure and by factors inherent to the insect . These toxins contain distinct domains that appear to be functionally important in toxin binding to protein receptors in the midgut of susceptible insects, and the subsequent formation of a pore in the insect midgut epithelium . In this article features necessary for the insecticidal activity of these toxins are discussed . These include toxin structure, toxin processing in the insect midgut, the identification of toxin receptors in susceptible insects, and toxin pore formation in midgut cells . In addition a number of B . thuringiensis toxins act synergistically to exert their full insecticidal activity . This synergistic action is critical not only for expressing the insecticidal activity of these toxins, but could also play a role in delaying the onset of insect resistance.

Mem Inst Oswaldo Cruz, 1995 Jan-Feb, 90(1), 65 - 8
Multilocus enzyme electrophoresis study of Bacillus sphaericus; Zahner V et al.; Multilocus enzyme electrophoresis (MLEE) has been used in the study of some Bacillus species . In this work we applied MLEE and numerical analysis in the study of the Bacillus sphaericus group . B . sphaericus can be distinguished from other entomopathogenic Bacillus by a unique allele (NP-4) . Within the species, all insect pathogens were recovered in the same phenetic cluster and all of these strains have the same band position (electrophoresis migration) on the agarose gel (ADH-2) . The entomopathogenic group of B . sphaericus seems to be a clonal population, having two widespread frequent genotypes (zymovar 59 and zymovar 119).

Mem Inst Oswaldo Cruz, 1995 Jan-Feb, 90(1), 135 - 8
Bacillus thuringiensis: fermentation process and risk assessment . A short review; Capalbo DM; Several factors make the local production of Bacillus thuringiensis (Bt) highly appropriate for pest control in developing nations . Bt can be cheaply produced on a wide variety of low cost, organic substrates . Local production results in considerable savings in hard currency which otherwise would be spent on importation of chemical and biological insecticides . The use of Bt in Brazil has been limited in comparison with chemical insecticides . Although Bt is imported, some Brazilian researchers have been working on its development and production . Fermentation processes (submerged and semi-solid) were applied, using by-products from agro-industries . As the semi-solid fermentation process demonstrated to be interesting for Bt endotoxins production, it could be adopted for small scale local production . Although promising results had been achieved, national products have not been registered due to the absence of a specific legislation for biological products . Effective actions are being developed in order to solve this gap . Regardless of the biocontrol agents being considered atoxic and harmless to the environment, information related to direct and indirect effects of microbials are still insufficient in many cases . The risk analysis of the use of microbial control agents is of upmost importance nowadays, and is also discussed.

Mem Inst Oswaldo Cruz, 1995 Jan-Feb, 90(1), 131 - 3
Biological control program against simuliids in the State of São Paulo, Brazil; Araujo-Coutinho CJ; In Brazil, the use of biological vector-control methods has been largely confined to experimental research, with little or no application of such techniques by public institutions responsible for implementing control programs . The notable exceptions have been the black fly control program carried out by the Health Secretariat in the State of Sao Paulo . Since the 1980s, Sao Paulo's "Superintendencia de Controle de Endemias" has been conducting studies on the viability of using Bacillus thuringiensis (H-14) for simuliid control, and the results have been so encouraging that the agency has now incorporated this method into its Simuliid Control Program.

Mem Inst Oswaldo Cruz, 1995 Jan-Feb, 90(1), 125 - 9
Bacillus sphaericus mosquito pathogens in the aquatic environment; Yousten AA et al.; The fate of Bacillus sphaericus spores in the aquatic environment was investigated by suspending spores in dialysis bags in fresh and seawater . Spore viability was lost more rapidly in seawater . Neither B . sphaericus nor B . thuringiensis israelensis (B.t.i.) spores mixed with pond sediment appeared to attach to the sediment . However, rapid decrease in B.t.i . toxicity suggested attachment of parasporal bodies to sediment . B . sphaericus toxin settled more slowly and less completely . B . sphaericus spores fed to larvae of four aquatic invertebrates were mostly eliminated from the animal gut in less than one week . An exception was the cranefly (Tipula abdominalis) where spores persisted in the posterior gut for up to five weeks.

Mem Inst Oswaldo Cruz, 1995 Jan-Feb, 90(1), 121 - 4
Some environmental and biological factors influencing the activity of entomopathogenic Bacillus on mosquito larvae in Brazil; Consoli RA et al.; The influence of environmental and biological factors on the efficacy of Bacillus thuringiensis serovar israelensis and B . sphaericus as mosquito larvicides are reviewed . The importance of strain dependence, cultivating media/methods, mosquito species/specificity, formulations and their relation to mosquito feeding habits, as well as temperature, solar exposure, larval density and concomitant presence of other aquatic organisms are addressed with reference to the present status of knowledge in Brazil.

Mem Inst Oswaldo Cruz, 1995 Jan-Feb, 90(1), 115 - 9
Integrated control measures against Culex quinquefasciatus, the vector of filariasis in Recife; Regis L et al.; Integrated control measures against Culex quinquefasciastus have been implemented in a pilot urban area in Recife, Brazil . About 3,000 breeding sites found within the operational area were responsible for very high mosquito densities recorded during the pretrial period . Physical control measures have been applied to cess pits before starting a series of 37 treatments of the other sites with Bacillus sphaericus strain 2362, over 27 months . In spite of the difficulties due to environmental conditions, very significant reductions in preimaginal population of C . quinquefasciatus were achieved and, as a consequence, low adult mosquito densities were maintained for a relatively long period of time . Entomological and environmental data gathered in this pilot project can contribute to design an integrated mosquito control program in Recife city.

J Urol, 1995 Jan, 153(1), 53 - 6
Intravesical bacillus Calmette-Guerin for the treatment of superficial transitional cell carcinoma of the prostatic urethra in association with carcinoma of the bladder; Schellhammer PF et al.; Between 1981 and 1989, 83 male patients with stages Ta, Tis and T1 transitional cell carcinoma were treated with bacillus Calmette-Guerin (BCG) . Of 17 patients with carcinoma in situ of the prostatic urothelium 13 had identifiable prostatic ducts and periurethral ductal transitional cell carcinoma was identified in 7 . At a median followup of 64 months (range 29 to 90) 12 of 17 patients (70%) had a complete response in the prostatic urethra . Among the 10 patients with mucosal carcinoma without ductal involvement 8 responded as did 4 of the 7 with mucosal and ductal involvement . A total of 9 patients had persistent tumor or recurrence in the bladder or prostate . Two men had recurrence in the prostatic urethra and, due to age and co-morbidity, both were treated by transurethral resection and fulguration . Cystectomy was performed in the remaining 7 patients . Three of 31 patients (10%) whose prostate urethral biopsies were negative before BCG therapy had a positive biopsy afterwards . After treatment with BCG, the actuarial curves for cancer specific, progression-free and overall survivals showed no statistical difference between male patients with an initially positive or initially negative prostatic urethral biopsy . BCG is a reliable agent for initial therapy of superficial prostatic transitional cell carcinoma . Careful followup can identify persistent tumor, recurrences or progression that identifies patients for whom cystectomy is appropriate.

J Exp Biol, 1995 Jan, 198 ( Pt 1), 91 - 6
Bacillus thuringiensis delta-endotoxin and larval Manduca sexta midgut brush-border membrane vesicles act synergistically to cause very large increases in the conductance of planar lipid bilayers; Martin FG et al.; Brush-border membrane vesicles prepared from midguts of Manduca sexta larvae were incorporated into planar phospholipid bilayers . Addition of Bacillus thuringiensis delta-endotoxin to the buffered salt solutions bathing these bilayers resulted in large irreversible increases in conductance . At pH 9.6, the smallest toxin-dependent increase in bilayer conductance observed was 13 nS . Similar conductance increases were never observed in the absence of delta-endotoxin or in delta-endotoxin-treated bilayers not containing components of insect brush-border membranes.

Appl Environ Microbiol, 1995 Jan, 61(1), 98 - 102
Specific oligonucleotide primers for detection of lecithinase-positive Bacillus spp . by PCR; Schraft H et al.; An assay based on the PCR has been developed to facilitate detection and identification of Bacillus cereus in foods . Three primers for the PCR have been designed within the sequence for cereolysin AB, a cytolytic determinant that encodes lecithin-hydrolyzing and hemolytic activities of B . cereus . With the PCR and hybridization, the specificity of the primers was tested with 39 isolates of the B . cereus group, with 17 other Bacillus spp., and with 21 non-Bacillus strains . Results demonstrate a high specificity of the three oligonucleotides for isolates of the B . cereus group . With a combined PCR-hybridization assay, the detection limit for B . cereus in artificially contaminated milk was 1 CFU/ml of milk.

Appl Environ Microbiol, 1995 Jan, 61(1), 326 - 32
Purification and characterization of ferulate and p-coumarate decarboxylase from Bacillus pumilus; Degrassi G et al.; Bacillus pumilus PS213 isolated from bovine ruminal fluid was able to transform ferulic acid and p-coumaric acid to 4-vinylguaiacol and 4-vinylphenol, respectively, by nonoxidative decarboxylation . The enzyme responsible for this activity has been purified and characterized . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of crude extract from a culture induced by ferulic acid or p-coumaric acid shows three bands that are not present in the crude extract of an uninduced culture, while the purified enzyme shows a single band of 23 kDa; the molecular mass calculated by size exclusion chromatography is 45 kDa . Enzyme activity is optimal at 37 degrees C and pH 5.5 and is not enhanced by any cation . Kinetic studies indicated a Km of 1.03 mM and a Vmax of 0.19 mmol.min-1/mg.liter-1 for ferulic acid and a Km of 1.38 mM and a Vmax of 0.22 mmol.min-1/mg.liter-1 for p-coumaric acid.

Appl Environ Microbiol, 1995 Jan, 61(1), 170 - 4
Purification and characterization of alpha-L-arabinofuranosidase from Bacillus stearothermophilus T-6; Gilead S et al.; Bacillus stearothermophilus T-6 produced an alpha-L-arabinofuranosidase when grown in the presence of L-arabinose, sugar beet arabinan, or oat spelt xylan . At the end of a fermentation, about 40% of the activity was extracellular, and enzyme activity in the cell-free supernatant could reach 25 U/ml . The enzymatic activity in the supernatant was concentrated against polyethylene glycol 20000, and the enzyme was purified eightfold by anion-exchange and hydrophobic interaction chromatographies . The molecular weight of T-6 alpha-L-arabinofuranosidase was 256,000, and it consisted of four identical subunits as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration . The native enzyme had a pI of 6.5 and was most active at 70 degrees C and at pH 5.5 to 6.0 . Its thermostability at pH 7.0 was characterized by half-lives of 53, 15, and 1 h at 60, 65, and 70 degrees C, respectively . Kinetic experiments at 60 degrees C with p-nitrophenyl alpha-L-arabinofuranoside as a substrate gave a Vmax, a Km, and an activation energy of 749 U/mg, 0.42 mM, and 16.6 kcal/mol, (ca . 69.5 kJ/mol), respectively . The enzyme had no apparent requirement for cofactors, and its activity was strongly inhibited by 1 mM Hg2+ . T-6 alpha-L-arabinofuranosidase released L-arabinose from arabinan and had low activity on oat spelt xylan . The enzyme acted cooperatively with T-6 xylanase in hydrolyzing oat spelt xylan, and L-arabinose, xylose, and xylobiose were detected as the end reaction products.(ABSTRACT TRUNCATED AT 250 WORDS)

Appl Environ Microbiol, 1995 Jan, 61(1), 122 - 9
Isolation of extracellular 28- and 42-kilodalton beta-1,3-glucanases and comparison of three beta-1,3-glucanases produced by Bacillus circulans IAM1165; Aono R et al.; Bacillus circulans IAM1165 produces three major extracellular beta-1,3-glucanases (molecular masses, 28, 42, and 91 kDa) during the stationary phase of growth . The 28- and 42-kDa enzymes were purified to homogeneity from the culture supernatant in this study . The properties of these two enzymes were examined, together with those of the 91-kDa enzyme previously isolated . The enzymatic properties of the 28- and 42-kDa beta-1,3-glucanases closely resemble each other . The enzymes belong to a category of endo type 1,3-beta-D-glucan glucanohydrolases . The enzymes were active at pH 4.0 to 7.0 . The optimum temperature of the reactions was 60 degrees C when laminarin (a soluble beta-1,3-glucan) was used as the substrate at pH 7.0 . The enzymes hydrolyzed barley glucan and lichenan (beta-1,3-1,4-glucans) more effectively than laminarin . Of the three enzymes, the 42-kDa enzyme lysed fungal cell walls the most effectively.

Arch Bronconeumol, 1995 Jan, 31(1), 32 - 4
{Coexistence of bronchogenic carcinoma and active pulmonary tuberculosis}; Martinez Moragon E et al.; We describe 9 patients with coincident active tuberculosis of the lung and bronchogenic carcinoma who were all diagnosed within a 5-year period . All were men, with a mean age of 55 +/- 10 years . The principal symptom was toxic syndrome lasting 1-3 months and the most common X-ray sign was alveolar consolidation . Analysis of tissue samples revealed squamous carcinoma (4 cases), adenocarcinoma (4 cases, one of which was bronchioloalveolar) and non-small cell carcinoma (1 case) . Tumoral stage was often advanced: IV (44%) and III (33%) . The tuberculosis bacillus was isolated in sputum (100%) and in bronchial aspirate (50%); no in vitro resistance was observed . Three patients were treated with radical surgery and three with radiotherapy . Average survival was 3 months in this series.

J Invertebr Pathol, 1995 Jan, 65(1), 61 - 7
Germination of Bacillus thuringiensis spores in bacteriophagous nematodes (Nematoda: Rhabditida); Borgonie G et al.; During screening of Bacillus thuringiensis isolates for nematicidal activity it was observed that spores of B . thuringiensis germinated in the intestine of bacteriophagous nematodes in the presence of antibiotics . This phenomenon was studied more closely by scanning electron microscopy . The nematodes were fed with bacterial spore-crystal mixtures in axenic culture medium supplemented with tetracyclin and chloramphenicol . Germination of spores was rare but was more frequently observed in Panagrellus redivivus than in other nematode species investigated . Germination of spores in the nematode intestine resulted in the colonization of the entire nematode within 24 hr . Crude nematode tissue preparations supported germination and subsequent growth of B . thuringiensis spores and vegetative cells . The mechanism for the loss of antibiotic activity in the nematode intestine is unknown . Since B . thuringiensis requires a nutrient-rich environment for reproduction, e.g., a cadaver, bacteriophagous nematodes may serve as suitable hosts for B . thuringiensis.

J Invertebr Pathol, 1995 Jan, 65(1), 35 - 42
Interaction of the insecticidal crystal protein CryIA from Bacillus thuringiensis with amino acid transport into brush border membranes from Bombyx mori larval midgut; Parenti P et al.; The activities of three related Bacillus thuringiensis delta-endotoxins, designated CryIA(a), CryIA(b), and CryIA(c), as inhibitors of K(+)-dependent amino acid transport into membrane vesicles prepared from the anterior and posterior portions of Bombyx mori larval midgut were measured . Under experimental conditions similar to those occurring in vivo (membrane potential approximately -100 mV, inside negative; pH 7.2in/8.8out; an inwardly directed K+ gradient) CryIA(a) toxin produced a clear dose-dependent inhibitory effect on leucine uptake by both the anterior and the posterior gut membrane vesicles, giving half-maximal inhibition constants (IC50) of 2.6 +/- 0.3 and 2.1 +/- 0.2 microgram toxin/mg membrane protein, respectively . The other two delta-endotoxins were practically inactive . A dose-dependent inhibition of amino acid transport by CryIA(a) toxin was also observed on the carrier-mediated K(+)-independent component, i.e., the leucine-only form . This result strongly indicates that the activity of the K+/amino acid cotransporter is directly affected after binding of delta-endotoxin to the brush border membrane . When the extravesicular pH was lowered to pH 7.2, the interaction of CryIA(a) toxin with the brush border appeared more complex, as suggested by the Hill coefficient of the dose-response curves higher than 1 . In conclusion, our data indicate that (i) CryIA(a) toxin specifically inhibited K+/leucine symport along the length of the midgut; (ii) the interaction between cotransporters and toxin is affected by the pH of the medium; and (iii) the K+/leucine cotransporter or a strictly associated protein may serve as membrane receptor for CryIA(a) delta-endotoxin in the B . mori larval midgut.

Plant Mol Biol, 1995 Jan, 27(1), 91 - 104
Inheritance of gusA and neo genes in transgenic rice; Peng J et al.; Inheritance of foreign genes neo and gusA in rice (Oryza sativa L . cv . IR54 and Radon) has been investigated in three different primary (T0) transformants and their progeny plants . T0 plants were obtained by co-transforming protoplasts from two different rice suspension cultures with the neomycin phosphotransferase II gene {neo or aph (3') II} and the beta-glucuronidase gene (uidA or gusA) residing on separate chimeric plasmid constructs . The suspension cultures were derived from callus of immature embryos of indica variety IR54 and japonica variety Radon . One transgenic line of Radon (AR2) contained neo driven by the CaMV 35S promoter and gusA driven by the rice actin promoter . A second Radon line (R3) contained neo driven by the CaMV 35S promoter and gusA driven by a promoter of the rice tungro bacilliform virus . The third transgenic line, IR54-1, contained neo driven by the CaMV 35S promoter and gusA driven by the CaMV 35S . Inheritance of the transgenes in progeny of the transgenic rice was investigated by Southern blot analysis and enzyme assays . Southern blot analysis of genomic DNA showed that, regardless of copy numbers of the transgenes in the plant genome and the fact that the two transgenes resided on two different plasmids before transformation, the introduced gusA and neo genes were stably transmitted from one generation to another and co-inherited together in transgenic rice progeny plants derived from self-pollination . Analysis of GUS and NPT II activities in T1 to T2 plants provided evidence that inheritance of the gusA and neo genes was in a Mendelian fashion in one plant line (AR2), and in an irregular fashion in the two other plant lines (R3 and IR54-1) . Homozygous progeny plants expressing the gusA and neo genes were obtained in the T2 generation of AR2, but the homozygous state was not found in the other two lines of transgenic rice.

Nord Med, 1995, 110(2), 45 - 7
{Tuberculosis control in Scandinavia}; Romanus V et al.; Compared with the rest of the world, the situation regarding tuberculosis is favourable in the Nordic countries . In 1993, the incidence for persons born in respective country (per 100,000 of the population) was 4.1 in Denmark, 3.2 in Iceland and Sweden, 3.8 in Norway and 10.0 in Finland . The respective average figures for foreign-born residents were 6-14 times higher . Programmes for contending with tuberculosis have been uniform in the Nordic countries, with the exception of BCG (bacillus Calmette-Guerin) vaccination policy, though childhood tuberculosis is rare in the area . However, tuberculosis represents a manifest problem among the foreign-born population, first and foremost among the increasing number of refugees.

Clin Exp Immunol, 1995 Jan, 99(1), 82 - 9
Lysis of human macrophages by cytolytic CD4+ T cells fails to affect survival of intracellular Mycobacterium bovis-bacille Calmette-Guérin (BCG); Fazal N et al.; Human CD4+, mycobacteria-specific, cytolytic T cell clones were used to lyse BCG-infected macrophages, and the effect on the subsequent growth and viability of the organisms was examined . The survival of released bacteria following cell lysis was assessed by both 3H-uridine labelling and colony-forming unit (CFU) estimation . The results indicate that even when effective antigen-specific or lectin-mediated cytolysis of the infected macrophages was achieved, there was no evidence for a direct mycobactericidal effect on the intracellular bacteria . This remained the case even if the period of co-culture of T cells and macrophages was extended up to 48 h . Pretreatment of the macrophages with interferon-gamma (IFN-gamma) was not able to act together with T cell-mediated lysis to produce inhibition of mycobacterial growth.

J Fam Pract, 1995 Jan, 40(1), 76 - 80
BCG vaccination and the PPD test: what the clinician needs to know; Ciesielski SD; The resurgence in tuberculosis necessitates careful surveillance and sensitive detection of cases . The purified protein derivative (PPD) test is the foundation of tuberculosis control . Primary care clinicans are encountering increasing numbers of persons immigrating from countries in which BCG (bacille Calmette-Guerin) vaccination is common . Many health care providers believe that previous BCG vaccination usually results in a positive PPD test, and therefore consider BCG vaccination status when interpreting a positive result on the PPD test . All articles listed in MEDLINE that included BCG and PPD as key words, a total of 62, were reviewed . Articles published before computerization of the medical literature, a total of 35, were reviewed in Index Medicus . This literature review indicates that there is little relationship between BCG vaccination and PPD positivity, and that BCG vaccination status should not be considered in the interpretation of a positive PPD test.

Infect Immun, 1995 Jan, 63(1), 259 - 63
Promotion of cytotoxic T-cell generation in mixed leukocyte culture by phosphatidylinositol-specific phospholipase C from Bacillus thuringiensis; Rahman SM et al.; Phosphatidylinositol-specific phospholipase C (PIPLC) from Bacillus thuringiensis, which cleaves phosphatidylinositol or glycosylphosphatidylinositol on the external cell surface to generate a second messenger for intracellular signal transduction (S . Rahman et al., FEBS Lett . 303:193-196, 1992), was found to preferentially promote the generation of alloantigen-specific cytotoxic T lymphocytes in mixed leukocyte culture . PIPLC affected an early stage of cytotoxic T-lymphocyte activation in culture, and there was no evidence of any soluble cellular mediators of this PIPLC action . PIPLC neither enhanced overall cell proliferation nor noticeably promoted interleukin-2 and -4 production in mixed leukocyte culture . The relative population size of Ly-2+ T cells was increased, however, in a late mixed leukocyte culture with PIPLC . In addition, PIPLC enhanced an anti-CD3 monoclonal antibody-induced early increase in {Ca2+}i . These results suggest a new parasite (bacterium)-oriented mechanism for enhancing antigen-driven host cytotoxic T-lymphocyte immunity which does not include promotion of interleukin-2 production.

Curr Genet, 1995 Jan, 27(2), 135 - 41
Expression cloning, purification and characterization of a beta-1,4-galactanase from Aspergillus aculeatus; Christgau S et al.; Expression cloning has been used to isolate a cDNA encoding beta-1,4-galactanase from the filamentous fungus Aspergillus aculeatus . A cDNA library was prepared from mycelia, inserted in a yeast expression vector and transformed into Saccharomyces cerevisiae . Thirteen clones secreting galactanase activity were identified from a screening of approximately 2.5 x 10(4) yeast colonies . All clones expressed transcripts of the same galactanase gene . The cDNA was re-cloned in an Aspergillus expression vector and transformed into Aspergillus oryzae . The recombinant enzyme had a molecular weight of 44,000 Da, an isoelectric point of pH 2.85, a pH optimum of pH 4.0-4.5, and a temperature optimum of 45-65 degrees C, which is similar to values obtained for a beta-1,4-galactanase purified from A . aculeatus . The enzyme degraded unsubstituted galactan to galactose and galactobiose . The deduced primary sequence of the enzyme showed no apparent homology to any known enzyme, in accordance with this being the first reported beta-1,4-galactanase cDNA . However, the deduced amino-acid sequence of a Bacillus circulans DNA sequence containing an open reading frame (ORF) with no known function, showed 36% identity and 60% similarity to the galactanase amino-acid sequence.

Plant Foods Hum Nutr, 1995 Jan, 47(1), 39 - 47
Anti-hypertensive substances in fermented soybean, natto; Okamoto A et al.; Natto is a traditional Japanese fermented food made by fermenting boiled soy beans with Bacillus natto . Its contents of inhibitors against the angiotensin converting enzyme (ACE, EC3.4.15.1) were investigated . Relatively strong inhibitory activity (IC50: 0.4 mg/ml, 11.8 inhibition units/g natto) was detected in natto extracts and the inhibitory activity observed in the viscous fraction was more potent than in the bean extract . Two groups of inhibitors in the viscous material, high and low molecular weight inhibitors, were resolved by dialysis test . The inhibitor of high molecular weight was a protein with low IC50 value (0.12 mg/ml) . The two types of low molecular weight inhibitors were detected in ethanol extracts (IC50: 0.53 mg/ml and 0.95 mg/ml) and they were found to be stable over a wide range of pH and temperature up to 100 degrees C . They were different in the mode of ACE inhibition . One is competitive, and the other noncompetitive against the hydrolysis of Bz-Gly-His-Leu by ACE.

J Basic Microbiol, 1995, 35(2), 83 - 92
Phenol and cresol metabolism in Bacillus pumilus isolated from contaminated groundwater; Gunther K et al.; From an aquifier contaminated with phenolic compounds seven bacterial strains able to grow on phenol and several mono- and disubstituted alkylphenols as sole source of carbon and energy were isolated . Five isolates belong to the genus Pseudomonas, two to the genus Bacillus . The isolate most active in utilization of the applied xenobiotics was identified as Bacillus pumilus and used for the investigation of the degradation pathways in liquid cultures . Cells of this strain precultured on phenol were able to utilize para-cresol as sole carbon source via the oxidation of the methylsubstituent and intradiol ring cleavage of the resulting protocatechuic acid, whereas an intradiol ring fission of the intermediate 4-methylcatechol led to 4-methylmuconolactone as dead end-product . Cells precultured on meta- and ortho-cresol were able to utilize the respective compounds as sole carbon sources via 3-methylcatechol, which induced the following extradiol ring fission pathway . Cells precultured on phenol were able to cooxidize meta- as well as ortho-cresol to 3-methylcatechol, which was cleaved via an intradiol ring fission, finally leading to the dead end-product 2-methylmuconolactone.

Genet Eng (N Y), 1995, 17, 99 - 117
Structure, function and engineering of Bacillus thuringiensis toxins; Thompson MA et al.; Nature has provided potent insecticidal toxins as fermentation products of many Bacillus thuringiensis strains . Elucidation of structure-function relationships for this class of natural toxins is in its early stages . Both direct experimentation and application of theoretical structure-function principles emerging from the rapidly growing field of protein structure are accelerating understanding of these toxins . Coupled with the increasing demand for biologically sound pesticides, the benefits from engineering nature's toxins for improved performance set the stage for exciting, fast growth in discovery, characterization and commercialization of new active ingredients for biopesticides and transgenic plants.

Probl Tuberk, 1995, (2), 24 - 6
{Antibodies to Bacillus megaterium H glycoprotein in patients with lung diseases}; Autenshlius AI et al.; Enzyme immunoassay performed in patients with malignant and benign affections of the lungs determined antibodies (Abs) to glycoprotein isolated from saprophyte microorganism Bacilus megaterium H . In pulmonary tuberculosis and nonspecific pulmonary inflammation the above Abs occurred in patients with fibrous-proliferative changes and associated neoplastic conditions . In lung cancer the Abs were detected in well and moderately differentiated tumors . Nondifferentiated and poorly differentiated variants did not exhibit any rise in Abs to glycoprotein.

Appl Microbiol Biotechnol, 1995 Jan, 42(5), 697 - 702
Improved production of the insecticidal CryIVD protein in Bacillus thuringiensis using cryIA(c) promoters to express the gene for an associated 20-kDa protein; Wu D et al.; Previous studies have shown that a 20-kDa protein enhances production of the insecticidal CytA and CryIVA proteins of Bacillus thuringiensis in Escherichia coli as well as CytA production and crystal formation in B . thuringiensis . To determine whether the 20-kDa protein could enhance CryIVD production, an expression vector was constructed with the 20-kDa open-reading frame under control of cryIA(c) promoters and the cryIVD gene under control of its own promoter . Acrystalliferous cells of B . thuringiensis transformed with this plasmid, designated pWF53, produced large bitrapezoidal CryIVD crystals that averaged 1.3 x 0.92 x 0.31 microns, approximately fivefold larger in volume than wild-type CryIVD crystals, and 1.7 fold the volume of crystals produced using the cryIVD operon, which contains the cryIVD gene and the gene for the 20-kDa protein . These results demonstrate that the 20-kDa protein significantly improves net synthesis of CryIVD and promotes CryIVD crystal formation . Improved production of proteins as diverse as CryIVD and CytA by the 20-kDa protein indicates this protein may be useful in facilitating the production of other proteins.

Curr Microbiol, 1995 Jan, 30(1), 45 - 7
The level of enzymes involved in the allantoin metabolism of Bacillus fastidiosus grown under different conditions; Muruke MS et al.; Bacillus fastidiosus was cultivated in batch and continuous culture on various carbon and nitrogen sources . The enzymes involved in allantoin degradation (allantoinase, urease, carboligase) of B . fastidiosus were hardly affected by either carbon or nitrogen source . In contrast, the enzymes involved in glycerol utilization (glycerol kinase, glycerol 3-phosphate dehydrogenase) were induced during growth on glycerol, but were not affected by the amount of allantoin present.

PDA J Pharm Sci Technol, 1995 Jan-Feb, 49(1), 50 - 2
Effect of 24 hour room temperature hold time on the heat resistance of Bacillus coagulans spores suspended in 10% Calcium Gluconate, USP; Rubio SL et al.; In an industrial application, biological indicator test units can often remain at room temperature for several hours prior to a steam sterilization validation cycle . A study was performed using the biological indicator Bacillus coagulans suspended in 10% Calcium Gluconate, USP, to determine if room temperature hold time had an effect on the resistance of the biological indicator . The results indicated that no significant change in resistance was seen for hold times up to 24 hours at room temperature.

PDA J Pharm Sci Technol, 1995 Jan-Feb, 49(1), 29 - 31
Effect of rubber stopper composition, preservative pretreatment and rinse water temperature on the moist heat resistance of Bacillus stearothermophilus ATCC 12980; Rubio SL et al.; Bacillus stearothermophilus spores (liquid suspension) were inoculated onto rubber stoppers and exposed to sublethal steam sterilization cycles at 120 degrees C . The D-values were determined using the fraction-negative method . An increase in heat resistance (D-value) of 200%-400% was observed when the spore suspension was inoculated onto rubber stoppers . The D-values ranged from 4.90-6.96 minutes 120 degrees C . No significant effect was seen when different preservatives were added to the stoppers nor when hot or cold rinse water temperatures were used after processing.

Br J Dermatol, 1995 Jan, 132(1), 113 - 5
Bacillary angiomatosis with cutaneous and oral lesions in an HIV-infected patient from the U.K; Levell NJ et al.; We describe an HIV positive patient with histologically confirmed cutaneous and oral lesions of bacillary angiomatosis, an unusual clinical presentation . As far as we are aware, this is the first reported case of cutaneous bacillary angiomatosis in the U.K.

Schweiz Monatsschr Zahnmed, 1995, 105(4), 455 - 60
{The Chemiklaven 7000/8000 on the test bench}; Guggenheim B; Sterilization of metal instruments in Chemiclaves is undoubtedly the most protective mode of sterilization, because corrosion and formation of spots is minimal . In the past sterilization with Chemiclaves was criticised, because of the emission of formaldehyde by older models and the questionable efficacy of formaldehyde sterilization for wrapped instruments . In the present investigation the capability of the newest Chemiclaves (7000/8000) to sterilize dental instruments was investigated . To monitor sterilization efficacy filter paper strips loaded with spores of Bacillus stearothermophilus or dental instruments directly contaminated with these spores were used as biological indicators . The performance of the Chemiclaves was assessed with and without a full load of instruments; after running half of the program cycle, the spore strips in the chamber being either unpacked or packed into paper bags . In addition it was investigated, whether it was feasible to use a formaldehyde free sterilization solution (Steri-Vap) with these sterilizers . The daily performance of the Chemiclaves in a dental practice and the emission of formaldehyde were tested in a room < 10 m3, in which both apparatus were operated at the same time . The Chemiclaves 7000/8000 sterilized reliably under the set test condition, however, the formaldehyde-free sterilization solution did not lead to an acceptable efficacy of sterilization . The new Chemiclaves efficiently adsorbed formaldehyde vapours . Harvey sterilizers require a meticulous observation of the producer's instruction, because in comparison to saturated steam the chemical vapours have a lower heat capacity . The Chemiclaves 7000/8000 are recommended for the sterilization of well cleaned, dry instruments wrapped in original MDT paper bags.

Eur Urol, 1995, 27 Suppl 1, 9 - 12
BCG treatment of existing Ta, T1 tumours or carcinoma in situ of the bladder; Akaza H; There are theoretical limits to the efficacy of intravesical instillation of anticancer agents as a method to prevent recurrence of superficial bladder cancer . Studies on the direct efficacy of intravesical instillation of bacillus Calmette-Guerin (BCG) to treat existing papillary tumours (Ta, T1) have shown a complete response rate of 66.4% and a partial response rate of 20.8% . These figures represent potent efficacy similar to that of BCG in the treatment of carcinoma in situ (CIS) . Moreover, this efficacy was found to persist over a long period of time, and the subsequent recurrence rate was markedly reduced . In cases with multiple tumours, or in subjects where concurrent CIS is strongly suspected, these findings suggest that intravesical instillation of BCG might best be performed prior to transurethral resection of bladder tumours.

Eur Urol, 1995, 27 Suppl 1, 23 - 8
Practical approaches to the prevention and treatment of adverse reactions to BCG; van der Meijden AP; Compared to intravesical chemotherapy, instillations with bacillus Calmette-Guerin (BCG) may provoke more frequently local and systemic reactions . As well as irritative bladder symptoms, BCG therapy may cause systemic side-effects, such as mild malaise, fever and, less commonly, life-threatening sepsis . It is important that risk factors for these adverse events are recognized, and that traumatic catheterization and instillations in inflamed or damaged bladders are avoided . Once recognized, virtually all side-effects can be treated successfully . Treatment options may include delaying or withholding BCG instillations, or using antipyretics and analgesics for reducing bladder symptoms . Treatment with tuberculostatic drugs for up to 6 months may be necessary in cases of severe systemic or local toxicity . In vitro, BCG is very susceptible to most antibiotics and tuberculostatic agents . Clinical trials are currently investigating whether the prophylactic use of tuberculostatic agents, such as isoniazid, or dose reduction in BCG can prevent adverse effects without impairing the antitumour efficacy of BCG.

Eur Urol, 1995, 27 Suppl 1, 2 - 8
BCG in perspective: advances in the treatment of superficial bladder cancer; Lamm DL; The management of superficial bladder cancer has advanced significantly in recent years . Controlled clinical trials suggest the benefits of cytotoxic chemotherapy apply primarily to well-differentiated tumours, are short-term and do not include a reduction in disease progression . In contrast, intravesical bacillus Calmette-Guerin (BCG) immunotherapy is effective in high-grade tumours, provides long-term protection from tumour recurrence and reduces disease progression . Controlled clinical trials have consistently demonstrated that BCG provides superior protection from tumour recurrence compared with thiotepa or doxorubicin . In comparisons with mitomycin C, only two of six studies found a significant advantage for BCG therapy; both studies evaluating high-risk patients showed a significant reduction in tumour recurrence with BCG compared with mitomycin C . Controlled trials have demonstrated that newer chemotherapies and more intensive regimens are not superior to standard thiotepa treatment . However, controlled comparisons of immunotherapy suggest that BCG is superior to both interferon and keyhole-limpet haemocyanin . Comparative studies showing BCG to be superior to chemotherapy and other immunotherapies have employed what we now know to be suboptimal BCG regimens . Using three additional weekly BCG (Connaught) treatments at 3 months, complete response in carcinoma in situ is increased from 73 to 87% . In patients who are disease free, maintenance BCG using three weekly treatments at 6-month intervals improves long-term disease-free status in stage Ta and T1 bladder cancer from 50 to 83% . Such maintenance therapy improves the excellent 86% survival obtained with a single 6-week course of BCG to 92% (p < 0.04).

Eur Urol, 1995, 27 Suppl 1, 19 - 22
Improving the efficacy of BCG immunotherapy by dose reduction; Pagano F et al.; Several clinical trials have shown intravesical bacillus Calmette-Guerin (BCG) to be effective in the prophylaxis of papillary tumour recurrences and in the therapy of bladder carcinoma in situ (CIS), as well as in delaying progression to muscle invasion . Nevertheless, the optimal regimen of BCG therapy for superficial bladder cancer has still to be defined . In a previous phase II trial, a low-dose regimen (BCG Pasteur strain, 75 mg) was able to achieve clinically significant response rates with a decrease in side-effects compared with other reported studies using standard-dose BCG . However, a phase III randomized trial--low dose versus standard dose (BCG Pasteur strain, 75 vs . 150 mg)--was considered necessary to clarify definitively the relationships between dose, efficacy and toxicity . The results of the interim analysis of 183 patients (performed in 1993) have shown response rates to be better in patients submitted to a low-dose BCG regimen (p = 0.0009) and a significant decrease in most of the common side-effects (cystitis, fever, haematuria; p < 0.05) . Breaking down the results by stage, no differences in response rates were found in patients with stage TaM (70 vs . 62% in low-dose and standard-dose regimens, respectively, p = 0.5) . In T1M and CIS stages, 82 and 0 (p = 0.07), and 64 and 0% (p = 0.0003) of patients were free of tumour following low-dose and standard-dose therapy, respectively . An additional 6-week course in patients who failed the induction course retrieved additional responses in both groups . No differences in progression rates were observed.(ABSTRACT TRUNCATED AT 250 WORDS)

Eur Urol, 1995, 27 Suppl 1, 13 - 8
Improving the safety of BCG immunotherapy by dose reduction . Cooperative Group CUETO; Martinez-Pineiro JA et al.; The preliminary results of a randomized, multicentre, prospective trial conducted by the Spanish Oncology Group (CUETO) are presented . The primary aim of the trial was to determine whether or not lowering the dose of bacillus Calmette-Guerin (BCG) could reduce toxicity without compromising efficacy . From February 1991 to June 1992, 500 patients with superficial bladder cancer (TaG2-3, T1G1-3) entered the trial . Of these patients, 252 were included in group A (BCG, 81 mg) and 248 in group B (BCG, 27 mg) . There were no significant differences in clinical and pathological characteristics between the two groups . Following treatment, significant differences arose between groups A and B in local severe toxicity (22.6 vs . 4.2%, respectively; p < 0.01) . Systemic toxicity was more common in group A than in group B (p < 0.01): pulmonary effects (2.3 vs . 0.4%), fever (26.9 vs . 12.9%) and malaise (16.2 vs . 8.4%), all p < 0.01 . Regarding efficacy, 204 patients in group A and 210 in group B were evaluated after a mean follow-up of 18.6 months . No significant differences in recurrence rate (18.1 vs . 19.5%) or progression rate (2.4 vs . 4.8%) were found . However, in patients with superficial bladder cancer with high risk of recurrence and progression (carcinoma in situ and G3), caution must be exercised in selecting the dose.

Trans R Soc Trop Med Hyg, 1995 Jan-Feb, 89(1), 37 - 40
Tuberculin sensitivity and HIV-1 status of patients attending a sexually transmitted diseases clinic in Lusaka, Zambia: a cross-sectional study; Duncan LE et al.; A cross-sectional study to estimate the prevalence of latent tuberculosis (TB) in a group of Zambians at high risk of human immunodeficiency virus type 1 (HIV-1) infection and to examine the effect of HIV-1 infection on the tuberculin response was conducted in the University Teaching Hospital in Lusaka, Zambia during July to September 1990 . Patients were selected from those presenting to the out-patient clinic for first referral with either sexually transmitted or skin disease . 268 adults were included in the study; 158 (59%; 95% confidence interval {CI} = 53-65%) were HIV-1 antibody positive . Of 82 HIV-1 negative participants who returned for Mantoux skin test reading, 51 (62%; 95% CI = 57-67%) had a positive test reaction (diameter > or = 10 mm) after receiving 2 units of RT-23 tuberculin . Of 106 HIV-1 positive participants who returned, only 32 (30%; 95% CI = 26-34%) had a diameter > or = 10 mm . Nine (28%) of the HIV-1 positive and Mantoux positive participants had large reactions > or = 30 mm, compared to 4 (8%) of the HIV-1 negative, Mantoux positive participants (P = 0.03) . Results in the HIV-1 negative group indicated a prevalence of latent TB of 62% in this population . HIV-1 infection was associated with a much higher frequency of negative response to tuberculin and with a few large skin test responses . Thus, in populations where HIV seropositivity is high, Mantoux skin tests cannot be used to assess those with latent TB who might benefit from chemoprophylaxisPIP: A cross-sectional study of the Mantoux response and HIV-1 status of a sample of patients with sexually transmitted diseases and skin diseases in Lusaka, Zambia, sought to estimate the prevalence of latent tuberculous infection . The sample was selected from patients attending the sexually transmitted diseases/dermatology section at the University Teaching Hospital, Lusaka, Zambia, between July and September 1990 . A questionnaire regarding socioeconomic factors, history of TB, contact with TB, location and documentation of bacillus Calmette-Guerin (BCG) scar(s) and history of BCG vaccination was completed, and a physical examination for acquired immune deficiency syndrome (AIDS) was carried out . The Mantoux result was recorded as the average diameter of induration, measured in 2 perpendicular directions by the pen and palpation method . A total of 158 patients (59%) were HIV-1 positive . Of the 66 women who took part, 46 (70%) were HIV-1 positive; of the 201 men, 112 (56%) were HIV-1 positive (p = 0.06) . 232 patients had sexually transmitted diseases, the commonest being genital ulceration; 123/231 (53%) were HIV-1 positive . The remaining 36 patients had skin diseases, the commonest being herpes zoster; 32/36 (89%) were HIV-1 positive . Of the 267 patients remaining in the study, 193 (72%) returned to have their Mantoux test read, 188 within 48-72 h . 106 (67%) HIV-1 positive patients and 82 (75%) HIV-1 negative patients returned . Of the 82 HIV-1 negative patients, 51 (62%) had a Mantoux reaction or= 10 mm; 55 (67%) had a reaction or= 5 mm . Of the 106 HIV-1 positive patients, only 32 (30%) had a Mantoux reaction or= 10 mm; 35 (33%) had a response or= 5 mm . Comparing HIV-1 negative and HIV-1 positive participants gave a significant odds ratio of 3.85 for a Mantoux response or= 10 mm . Among the individuals with a Mantoux reaction or= 10 mm, 9/32 (28%) of HIV-1 positive participants had a megareaction or= 30 mm, while megareactions occurred in 4/51 (8%) of HIV-1 negative participants (odds ratio 4.6) .

Eur Urol, 1995, 27(2), 89 - 95
Bacillus Calmette-Guérin in superficial bladder cancer: consensus and controversies; Vegt PD et al.; In this overview, Bacillus Calmette-Guerin (BCG) immunotherapy in superficial bladder cancer items are discussed on which consensus has been reached and on which controversies exist . The evaluation of the optimal route of administration has shown that intravesical instillation of BCG alone is accepted as the best route of administration . In searching for the appropriate BCG strain, the analysis of the results of 6 substrains has made clear that no particular strain has shown superiority over others . In finding the optimal treatment schedule there is strong evidence that maintenance therapy is superior to induction therapy alone . No consensus has been reached about the optimal dose for BCG therapy nor about how the toxicity of BCG treatment can be reduced . Although some reports have stated that BCG immunotherapy is superior to chemotherapy for the treatment of patients with superficial bladder cancer, more data are needed to prove this statement . In conclusion, although BCG has been proven to be very effective in the treatment of patients with superficial bladder cancer, it is certainly not a panacea for all patients with superficial bladder cancer.

Urol Res, 1995, 22(6), 373 - 6
Drug therapy of bacillus Calmette-Guérin sepsis; Koukol SC et al.; Intravesical bacillus Calmette-Guerin (BCG) is widely used for the treatment of transitional cell carcinoma of the bladder . Although it is usually well tolerated, sepsis can occur, which has resulted in at least eight deaths {3} . The survival of Connaught BCG-infected mice treated with single and combination antibiotic and steroid therapy was evaluated . Triple-drug therapy with isoniazid, rifampin, and prednisolone resulted in 53% survival compared with 25% survival in the control group (P = 0.0209) . A survival of only 10.5% was observed with treatment using prednisolone alone . This survival was worse than that of the control group (25%), and approached statistical significance (P = 0.0669) . Our data suggest that BCG sepsis probably has components of both a hypersensitivity reaction and bacterial sepsis; they support the current use of combination antibiotic and steroid therapy for treatment of BCG sepsis in humans, but argue against treatment with steroids alone.

Rev Med Interne, 1995, 16(3), 191 - 4
{Osteoarticular tuberculosis . Diagnostic contribution of local sampling}; Lemaitre F et al.; Bone and joint tuberculosis have recently gained a renewal of interest, especially with the spread of HIV infection which may increase its frequency . Bone and joint locations of tuberculosis are pauci-bacillary often requiring local sampling in order to confirm the diagnosis and to initiate early therapy . From 1983 to 1992 we have studied 19 patients with bone and joint tuberculosis . Seventeen local sampling were performed: 12 biopsies and five abscess punctures . Pathological examination of samples disclosed diagnosis of tuberculosis in eight cases out of 12 . Among the remaining four patients, direct smear was positive once, and cultures grew Mycobacterium tuberculosis in two, yielding the diagnosis in 11 out of the 12 patients . Bacteriological and pathological examinations were non contributive in only one patient . Microbiological examination of pus disclosed two positive direct smear and three positive cultures . Treatment lasted 9 to 18 months . The outcome was favourable in all patients.

Mikrobiol Z, 1995 Jan-Feb, 57(1), 3 - 13
{The distribution of bacteria of the genus Bacillus into clusters by their spectrum of antagonistic activity}; Smirnov VV et al.; The antagonistic activity of Bacillus strains against a lot of the pathogenic test-cultures was determined . It was established that strains-antagonists might be divided into several groups with a range of the most susceptible test-cultures typical of each group . The method of the rapid screening of strains-antagonists with the desirable antagonistic quality was suggested.

Clin Infect Dis, 1995 Jan, 20(1), 136 - 42
Bacille Calmette-Guérin vaccination for the prevention of tuberculosis in health care workers; Brewer TF et al.; For 60 years vaccination with bacille Calmette-Guerin (BCG) has been used for the prevention of tuberculosis in health care workers . In 1988 the U.S . Advisory Committee on Immunization Practices removed the category of health care worker from the list of persons for whom vaccination with BCG should be considered . Nosocomial epidemics of tuberculosis, especially those caused by multidrug-resistant strains, have led to the reconsideration of vaccination with BCG for this population . We review the available studies of the efficacy of BCG vaccine in health care workers . Although the studies had too many methodological flaws to be combined in a quantitative meta-analysis, they suggest that vaccination with BCG is effective in reducing the incidence of tuberculosis among health care workers.

Clin Infect Dis, 1995 Jan, 20(1), 126 - 35
Relationship between bacille Calmette-Guérin (BCG) strains and the efficacy of BCG vaccine in the prevention of tuberculosis; Brewer TF et al.; Bacille Calmette-Guerin (BCG) vaccination for the prevention of tuberculosis has been used in humans since 1921 . Furthermore, for > 60 years it has been possible to separate BCG strains (defined here as a BCG vaccine maintained in a particular laboratory and used in a particular trial or set of trials) on the basis of in vitro and in vivo tests . Investigators have concluded that differences in the BCG strains used in efficacy trials on humans may be responsible for the wide range in levels of protection from tuberculosis reported in those trials . We review the development of the separate strains used in the trials included in a recent meta-analysis and examine data for and against the protective efficacy of different BCG strains . The difficulties in correlating results of in vitro and in vivo tests with protective efficacy in humans are discussed . The limited data available from human studies suggest that the BCG strain used for vaccination is not a significant determinant of the overall efficacy in the prevention of tuberculosis.

Vet Pathol, 1995 Jan, 32(1), 63 - 5
Naturally occurring Tyzzer's disease in a puppy; Young JK et al.; A naturally occurring case of Tyzzer's disease due to infection with Bacillus piliformis in a wolf-dog hybrid resulted in widely disseminated lesions, including severe myocarditis, hepatitis, enterocolitis, intestinal leiomyositis, and adrenal cortical adenitis . Previously reported lesions for canine Tyzzer's disease have been limited to hepatic necrosis and a necrotizing enterocolitis.

Curr Opin Rheumatol, 1995 Jan, 7(1), 48 - 53
Musculoskeletal syndromes associated with malignancies; Seda H et al.; Literature on the association of malignancies with various rheumatic disorders published over the past year is summarized in this review . The possible roles of methotrexate treatment in predisposing to the development of lung cancer and Felty's syndrome in predisposing to non-Hodgkin's lymphoma in rheumatoid arthritis patients are discussed . The increased occurrence of monoclonal gammopathies and non-Hodgkin's lymphoma in patients with Sjogren's syndrome is reported . The possible increased frequency of malignancies among patients with systemic lupus erythematosus (SLE), scleroderma, and polymositis-dermatomyositis is revisited; of interest, the overlapping clinical features of non-Hodgkin's lymphoma and SLE are presented, as well as the increased occurrence of ovarian cancer in patients (especially older women) with dermatomyositis . The proceedings of the first International Workshop on Hypertrophic Osteoarthropathy, as well as the association of this syndrome with nasopharyngeal carcinoma in the childhood years, are presented . Finally, postchemotherapy and post-bacille Calmette-Guerin rheumatism are described.

Mikrobiologiia, 1995 Jan-Feb, 64(1), 18 - 22
{The effect of Bacillus intermedius RNAase on the growth and development of Bacillus mucilaginosus}; Kupriianova-Ashina FG et al.; The effect of Bacillus intermedius RNAase on vegetative cells and spores of siliceons bacteria Bacillus mucilaginosus was investigated . It is shown that the enzyme stimulates the growth of vegetative cells of B . mucilaginosus at a concentration of 10 mkg/ml and promotes germination of spores at a 1000-fold lesser concentration (0.1 mkg/ml) . The spores at stages of activation and initiation appear to be most susceptible to the enzyme action . The effect of RNAase on B . mucilaginosus multiplication correlates with intensification of bacterial leaching of bauxites.

Yao Xue Xue Bao, 1995, 30(1), 46 - 58
{Morphological and histological studies of the Chinese drug lao-guan-cao}; Zhang YY et al.; Lao-guan-cao has long been used in Chinese traditional medicine . It is used as an antirheumatic and circulation promoting drug for the treatment of acute and chronic rheumatalgia; also used as a detoxicant for enteritis and bacillary dysentery . The dried aerial parts of Erodium stephanianum Willd . and Geranium wilfordii Maxim . (family Geraniaceae) have been specified in the Chinese Pharmacopoeia (1990 ed.) According to the literature and the authors' market investigations, the commercial drug Lao-guan-cao chiefly consists of the aerial parts of Erodium stephanianum Willd . Also those rather popularly used in several provinces are the aerial parts of Geranium nepalense Sweet and G . sibiricum L . In this paper, the morphological and histological characters of the crude drugs derived from E . stephanianum Willd., G.nepalense Sweet and G.sibiricum L . are described . It is found that the following characters are useful for distinguishing the drugs derived from the above mentioned three species . (1) The shape and size of leaf; (2) the number and arrangement of the vascular bundles of stem; (3) the number of the vascular bundles in midrib and layers of palisade cells of leaf; (4) the presence or absence of stomata in upper epidermis; (5) the characters of cells containing crystals in sepal; (6) the characters of epidermis cells, cells containing crystals and fibers of pericarp; (7) the types of hairs and glandular hairs on epidermis of stem, leaf, sepal and pericarp; (8) the characters of anticlinal walls of epidermal cells of seed coat.

Arch Virol, 1995, 140(7), 1173 - 80
Ultrastructure and immuno-cytochemistry of BHK-21 cells infected with a modified Bucyrus strain of equine arteritis virus; Wada R et al.; Morphogenesis of a modified Bucyrus strain of equine arteritis virus (EAV) in BHK-21 cells was studied . Bacillary tubules were first detected in the cytoplasm 8 h after infection, and mature virions 79 to 122 nm in diameter, 101 nm on average, were mostly observed in the cisternae of the rough endoplasmic reticulum (RER) at 12 h or later . They had isometrical cores and morphological subunits in the outer layer . Budding occurred from the RER and the outer nuclear membrane, but not from the cell surface . Structural linkage was detected between the tubule and the virus core . Aberrant strands were occasionally demonstrated within the nucleus 12 h after infection, and immunofluorescence and immunogold labeling revealed viral antigen also in the nucleus.

Can J Infect Control, 1995 Summer, 10(2), 41 - 4
Tuberculin skin testing in a hospital and two chronic care facilities in Prince Edward Island; Langille DB et al.; OBJECTIVE: To determine the prevalence of initial and booster positive responses to tuberculin skin testing among staff of an acute care hospital, and among staff and residents of two chronic care facilities on Prince Edward Island (PEI) . PATIENTS AND METHODS: Two hundred and eighty-six staff from the hospital and the facilities, and 164 residents from the facilities were given tuberculin testing using 5 tuberculin units (TU) of purified protein derivative (PPD) of tuberculin . A two-step booster test was performed on 125 of 173 staff (73%) aged 35 years or older who initially tested negative, and on 138 of 141 residents (97.9%) at the chronic care homes . RESULTS: The overall rate of initial tuberculin positivity among staff at the facilities was 15.7% . Among residents, the rate of positivity was 14.0% . Histories given by institutional staff of having previously received Bacille bilie de Calmette-Guerin vaccine (BCG) were associated with tuberculosis (TB) test reactivity, but not with mean reaction size . Staff with a history of BCG who tested positive did not differ in age from those testing negative . Institutional staff whose work involved patient contact were more likely to have a positive test than those whose work did not involve such contact . Booster positivity among those older than 35 years of age ranged from 2.0 to 5.2% . CONCLUSIONS: These results suggest that, for health care institutions in PEI, tuberculin testing should be performed on all new or current hospital staff at risk of TB exposure, all new or current staff of chronic care facilities, and all new or current residents of chronic care facilities unless contraindicated . Booster testing should also be performed on all institutional staff and residents 35 years of age or older if their initial tuberculin test is negative.

Can J Infect Control, 1995 Summer, 10(2), 37 - 40
Bacillus species pseudobacteremia following hospital construction; Loeb M et al.; A 13-fold increase (0.08 to 1.05%) in the isolation rate of Bacillus species from in-patient blood cultures led to the investigation of pseudobacteremia over a four-month period . Data on blood isolates (BACTEC NR 660, bottle types 6A and PEDS) of Bacillus species were compiled between July and October 1993 . BACTEC bottles, as well as their plastic lids and rubber septa (representing 20% of stored bottles), and alcohol swabs, iodine preparation pads and butterfly collection systems were cultured . Air plates inside the BACTEC 660 instrument were cultured, as well as swabs of both the BACTEC needles and the interior of the BACTEC handling area . To investigate carry-over contamination, sterile BACTEC bottles were tested immediately following bottles containing Bacillus species . Growth of Bacillus species was obtained from 16% of the plastic lids and 1.3% of the rubber septa . No growth was obtained from other cultures . The outbreak coincide with construction on a driveway of the hospital over the area where BACTEC bottles were stored . Upon completion of construction in November 1993, the isolation of Bacillus species returned to baseline . No pseudobacteremia isolates occurred in areas where the trained intravenous team was assigned . In conclusion, hospital construction leading to airborne spread of Bacillus species may cause Bacillus species pseudobacteremia, and failure to disinfect blood culture bottles adequately may predispose to such an outcome.

Urol Int, 1995, 54(3), 137 - 41
Evaluation of cellular immunity following bacillus Calmette-Guérin therapy in patients with superficial bladder cancer; Sarica K et al.; Despite its marked effectiveness in the prevention of tumor recurrences, a great deal of information on the mode of action of Bacillus Calmette-Guerin (BCG) as an antitumor therapy modality is still lacking . In this prospective study, by performing lymphocyte subset analysis and quantitative assessment of delayed hypersensitivity skin reactions before transurethral resection of the detected tumor and 3-6 months after intravesical BCG administration in 23 patients with superficial bladder carcinoma, we tried to demonstrate the immunostimulatory effect of BCG therapy . We had 4 recurrences at 6 months' follow-up . Evaluation of lymphocyte subset analysis readings in our group revealed a statistically significant difference (p < 0.05) in CD4+/CD8+ ratio between baseline values and that obtained following BCG administration at 3 and 6 months . However, there was no statistically significant increase of this value in 4 patients who had tumor recurrences . Evaluation of delayed hypersensitivity skin test score results revealed a statistically significant increase in the whole group at 3 months of follow-up (p < 0.05) but the same evaluation at 6 months of follow-up showed no statistically significant difference with respect to this evaluation . Again, no statistically significant difference was found in 4 patients who had tumor recurrences . These results support the idea that BCG-associated antitumor activity is an immune-mediated reaction and the assessment of T-lymphocyte subsets together with quantitative evaluation of delayed hypersensitivity skin reactions would give us a definite idea about the immunotherapeutic effects of BCG in such patients.

Rev Inst Med Trop Sao Paulo, 1995 Jan-Feb, 37(1), 7 - 12
Pattern of mycobacterial antigen detection in leprosy; Pagliari C et al.; Immunohistochemistry reaction (Peroxidase anti-peroxidase--PAP) was carried out on fifty-two skin biopsies from leprosy patients with the purpose to identify the antigenic pattern in mycobacteria and to study the sensitivity of this method . Five different patterns were found: bacillar, granular, vesicular, cytoplasmatic and deposits, classified according to the antigenic material characteristics . Deposits (thinely particulate material) appeared more frequently, confirming the immunohistochemistry sensitivity to detect small amounts of antigens even when this material is not detected by histochemical stainings.

Rev Argent Microbiol, 1995 Jan-Mar, 27(1), 21 - 7
{Bacteria of the genus Bacillus in chicken carcasses and hamburgers}; Iacona VA et al.; The incidence of mesophilic aerobic sporulate bacteria in chicken carcasses and hamburgers was studied and the species of isolated sporulate bacteria were identified . Forty seven eviscerated carcasses from a processing plant of Entre Rios province (Argentina) were analyzed together with fifty samples of hamburgers from two supermarkets of Santa Fe city . All carcasses resulted in contamination with aerobic mesophilic bacteria in the range from 6 x 10(3) to 1.2 x 10(6) CFU/ml liquid washed, and 94% them with sporulate bacteria, the threshold being under 100 CFU/ml (Figure 1) . Hamburgers from both places resulted with aerobic mesophilic bacteria in 100% of the cases, in the range of 1 x 10(5)-3.3 x 10(6) CFU/g for supermarket A and 2.2 x 10(5) to 1.7 x 10(7) CFU/g for supermarket B; the incidence of sporulate bacteria was between 4.3 x 10(2) and 1.2 x 10(4) CFU/g for A, while the range for B was 6.2 x 10(2) and 3.8 x 10(4) CFU/g (Figure 2) . Two hundred and fourteen Bacillus Genus strains were isolated and purified from the carcasses and five hundred and ninety five from hamburgers . B . subtilis and B . megaterium were most involved in carcasses; while B . licheniformis, B . subtilis and B . pumilus were in hamburgers from supermarket A and B . subtilis and B . pumilus were found in supermarket B (Table 1) . The presence of B . cereus was also found, although in low levels, in all the samples (Table 1) . Pollution levels with aerobic mesophilic bacteria are high in both kinds of samples.(ABSTRACT TRUNCATED AT 250 WORDS)

Virchows Arch, 1995, 427(1), 85 - 9
Identification of mycobacteria to the species level by automated restriction enzyme fragment length polymorphism analysis; Totsch M et al.; An automated method for the restriction fragment length polymorphism (RFLP) analysis for the differentiation of mycobacteria to the species level is described . After polymerase chain reaction (PCR) amplification of a sequence of the gene encoding the 65-kDa surface antigen common to all mycobacteria the product was investigated by RFLP analysis . For accurate determination of fragment sizes the asymmetrically fluorescein-labelled PCR product was partially digested with restriction site enzymes BstEII and HaeIII . The fragments obtained were analysed electrophoretically using an automated laser fluorescence DNA sequencer . Determination of fragment sizes revealed a deviation of +/- 1 base pair (bp; 0.6%) when compared to expected sizes . The validity of this approach was confirmed by analysing mycobacterial DNA obtained from pure cultures of Mycobacterium (M.) tuberculosis and alcohol-fixed smears as well as paraffin-embedded sputa of patients with culture-proven tuberculosis . Additionally a diagnostic algorithm was established by investigation of cultured M . bovis, M . bovis bacille Calmette-Guerin, M . avium, M . intracellulare and M . fortuitum . The method allows the identification of restriction enzyme sites which are only 40 bp apart . Partial restriction enzyme digestion of asymmetrically fluorescence-labelled PCR products will presumably lead to the discovery of new restriction enzyme sites.

Eur Urol, 1995, 27(4), 334 - 8
Late effects of intravesical bacillus Calmette-Guérin immunotherapy on bladder mucosa infiltrating lymphocytes: an immunohistochemical study; Boccafoschi C et al.; During intravesical bacillus Calmette-Guerin (BCG) treatment for the prophylaxis of recurrent superficial bladder carcinoma, patients typically show a local inflammatory response involving mainly T lymphocytes, most of which have the helper-induced phenotype (CD4+) (CD4+/CD8+ ratio > 1) . To evaluate whether this immunophenotypic profile of the lymphocytes persists also after the completion of this immunotherapy, we examined bladder biopsy specimens during the posttreatment follow-up period of 24 patients, previously submitted to a 2-year BCG administration . The intensity of inflammatory response differed among the patients and in 10 of them even between the scar and the normal mucosa of the bladder . A reversal to the pretreatment CD4+/CD8+ ratio < 1 occurred in the majority of subjects, including the 3 patients with histologically confirmed tumour recurrence . In addition, 11 tumour-free patients showed prevailing CD4+ cells in the scar mucosa and prevailing CD8+ in the normal mucosa of their bladder or vice versa . From these findings it appears that the long-term host response to BCG does not depend exclusively on an intense, long-lasting local mononuclear immune reaction.

Hautarzt, 1995 Jan, 46(1), 39 - 43
{Bacillary angiomatosis}; Plettenberg A et al.; Bacillary angiomatosis (BA) is a rare infectious disease usually associated with HIV infection . Recent molecular biologic investigations confirm that both Rochalimaea henselae and Rochalimaea quintana can cause BA . The bacteria can be identified by Warthin-Starry staining and electron microscopy . The typical clinical signs are solitary or multiple dermal or subcutaneous nodules . Bone, liver, spleen and other organs may also be involved . We describe the clinical and histological features of a 39-year-old HIV-infected patient with cutaneous and bony lesions of BA . All manifestations of BA disappeared during therapy with erythromycin.

Microbiol Immunol, 1995, 39(8), 619 - 22
Monoclonal antibody developed against a hemolysin of Bacillus thuringiensis; Matsuyama J et al.; A total of five hybridoma cell lines that produced monoclonal antibodies (MAb) against a hemolysin (Bt-hemolysin) produced by Bacillus thuringiensis were established and characterized . All of these monoclonal antibodies reacted similarly not only to Bt-hemolysin but also to a hemolysin (Bc-hemolysin) produced by B . cereus, suggesting that the two hemolysins are immunologically indistinguishable . The MAb developed in this study was also successfully applied for rapid and simple purification of both Bt- and Bc-hemolysins by immunoaffinity column chromatography . The partial N-terminal amino acid sequence of the purified hemolysins was determined to be Ile-Glu-Gln-Thr.

Rocz Panstw Zakl Hig, 1995, 46(1), 91 - 7
{The influence of diesel fuel oil on the number of bacteria, fungi, actinomycetes and soil microbial biomass}; Michalcewicz W; In laboratory conditions the influence of Diesel fuel oil on the total number bacteria, fungi, actinomycetes and content of microbial biomass in soil was studied . The fuel oil was applied at dosage of 1, 2 and 5 g/100 g of soil . Samples of soil were analysed 1, 7, 14, 21 and 150 days after oil applied . Microbiological analyses involved determinations of the total number of bacteria and actinomycetes (on Bunta-Roviry medium) and fungi (on Martin's agar) by the plate method . The content of microbial biomass in soil was determined using the physiological method of Anderson and Domsch . Diesel fuel oil application caused the changes of total number of investigated groups of soil microorganisms . These results were confirmed by the investigated groups of soil microorganisms . These results were confirmed by the investigation on microbial biomass in soil . In the most of cases the fuel oil was the most stimulated for the number of bacteria and the most inhibited for actinomycetes in comparison with soil without oil (control) . From soil with Diesel fuel oil addition Pseudomonas, Arhtrobacter, Bacillus and Penicillium, Aspergillus, Fusarium, Trichoderma strains were isolated . The possibility of the fuel oil as source of carbon utilization by these strains will be investigate.

Biochemistry, 1994 Dec 27, 33(51), 15408 - 17
Crystal structure of the ferredoxin I from Desulfovibrio africanus at 2.3 A resolution; Sery A et al.; The crystal structure of the ferredoxin I from the sulfate-reducing bacterium Desulfovibrio africanus (DaFdI) has been solved and refined by X-ray diffraction . The crystals are orthorhombic with a = 96.6 A, b = 58.1 A, and c = 20.7 A, space group P2(1)2(1)2, and two ferredoxin molecules per asymmetric unit . The initial electron density map has been obtained by combining phasing by molecular replacement methods, anomalous scattering, and noncrystallographic averaging . The final crystallographic R factor is 0.182 with 10-2.3 A resolution data . In parallel, the amino acid sequence was redetermined . This showed that DaFdI contains 64 residues (instead of 61) including one free cysteine, one histidine, and one tryptophan in the C-terminal part of the molecule . The current molecular model includes the two molecules of the asymmetric unit, 67 water molecules, and one sulfate ion . The DaFdI overall folding very closely resembles that of ferredoxins of known structure . Comparisons with the single cluster ferredoxins from Desulfovibrio gigas and Bacillus thermoproteolyticus show that the presence or the absence of a disulfide bridge does not significantly affect the folding of the other half of the molecule, including the characteristic alpha-helix of the single cluster ferreddoxins . Like other ferredoxins or analogs, the {4Fe-4S} iron--sulfur cluster presents, at 2.3 A resolution, a cubane-like geometry . By contrast, its immediate environment is different as it includes, besides the four cysteic sulfur ligands, the sulfur atom of the free cysteine . This sulfur atom, which is buried within the protein, is in van der Waals contact with one labile sulfur of the cluster and one liganded cysteic sulfur . The association of a {4Fe-4S} cluster with one free cysteic sulfur is similar to that previously found in both X-ray structures of Azotobacter vinelandii and Peptococcus aerogenes {Stout, C . D . (1989) J . Mol . Biol . 205, 545-555; Backes, G., et al . (1991) J . Am . Chem . Soc . 113, 2055-2064} . Chemical sequence analysis suggests that this characteristic {4Fe-4S} cluster sulfur environment is widely distributed among ferredoxins.

FEMS Microbiol Lett, 1994 Dec 15, 124(3), 271 - 5
Role of the exosporium in the stability of the Bacillus sphaericus binary toxin; Nicolas L et al.; The persistence of toxicity of the Bacillus sphaericus 1593 binary toxin was compared when produced in B . sphaericus, inside the exosporium, or in a recombinant B . thuringiensis strain, outside the exosporium . The stability of the toxin crystal was affected by temperature and quality of the water, but not by the location of the production in the bacterial cell.

Biochem J, 1994 Dec 15, 304 ( Pt 3), 895 - 902
The alpha-5 segment of Bacillus thuringiensis delta-endotoxin: in vitro activity, ion channel formation and molecular modelling; Gazit E et al.; A peptide with a sequence corresponding to the highly conserved alpha-5 segment of the Cry delta-endotoxin family (amino acids 193-215 of Bacillus thuringiensis CryIIIA {Gazit and Shai (1993) Biochemistry 32, 3429-3436}), was investigated with respect to its interaction with insect membranes, cytotoxicity in vitro towards Spodoptera frugiperda (Sf-9) cells, and its propensity to form ion channels in planar lipid membranes (PLMs) . Selectively labelled analogues of alpha-5 at either the N-terminal amino acid or the epsilon-amine of its lysine, were used to monitor the interaction of the peptides with insect membranes . The fluorescent emission spectra of the 7-nitrobenz-2-oxa-1,3-diazole-4-yl (NBD)-labelled alpha-5 peptides displayed a blue shift upon binding to insect (Spodoptera littoralis) mid-gut membranes, reflecting the relocation of the fluorescent probes to an environment of increased apolarity, i.e . within the lipidic constituent of the membrane . Moreover, midgut membrane-bound NBD-labelled alpha-5 peptides were protected from enzymic proteolysis . Functional characterization of alpha-5 has revealed that it is cytotoxic to Sf-9 insect cells, and that it forms ion channels in PLMs with conductances ranging from 30 to 1000 pS . A proline-substituted analogue of alpha-5 is less cytolytic and slightly more exposed to enzymic digestion . Molecular modelling utilizing simulated annealing via molecular dynamics suggests that a transbilayer pore may be formed by alpha-5 monomers that assemble to form a left-handed coiled coil of approximately parallel helices . These findings further support a role for alpha-5 in the toxic mechanism of delta-endotoxins, and assign alpha-5 as one of the transmembrane helices which form the toxic pore . The suggested role is consistent with the recent finding that cleavage of CryIVB delta-endotoxin in a loop between alpha-5 and alpha-6 is highly important for its larvicidal activity {Angsuthanasombat, Crickmore and Ellar (1993) FEMS Microbiol . Lett . 111, 255-262}.

Biochemistry, 1994 Dec 13, 33(49), 14858 - 70
1H, 13C, and 15N resonance assignments and secondary structure analysis of the HU protein from Bacillus stearothermophilus using two- and three-dimensional double- and triple-resonance heteronuclear magnetic resonance spectroscopy; Vis H et al.; Nearly complete 1H, 13C, and 15N resonance assignments have been obtained for the protein HU from Bacillus stearothermophilus (dimer, 19.5 kDa) using double- and triple-resonance 2D and 3D NMR experiments . This has resulted in assignments of 91% of the observable protons, 98% of all 13C, and 92% of all 15N nuclei . NOEs obtained from a 3D time-shared NOESY-(13C,15N)-HSQC spectrum, exchange data of amide protons, and chemical shifts of the 1H alpha, 1HN, 13C beta, 13C alpha, 13CO, and 15N nuclei have been used to identify the secondary structure elements . Three alpha-helices (residues 3-13, 18-37, and 83-90) and three extended strands (residues 40-45, 48-62, and 67-82) have been found in HU . The arrangement of these elements of secondary structure is very similar to the X-ray structure {Tanaka et al . (1984) Nature 310, 376-381; White et al . (1989) Proteins 5, 281-288} . The conformation of the proposed DNA-binding region of HU, i.e., an antiparallel beta-hairpin, was not observed previously in the X-ray structure . In the NMR structure long range NOEs in the beta-arm region (residues 53-76) suggest a distortion between residue Pro-72 and Ala-73 and between Pro-63 and Gln-64 with concomitant distortions in the opposite strand . The NOE data indicate further that the loop region in the DNA-binding arms of HU is arranged as a type I beta-turn from Pro-63 to Gly-66.

FEBS Lett, 1994 Dec 12, 356(1), 30 - 2
Biochemical approaches of increasing thermostability of beta-amylase from Bacillus megaterium B6; Ray RR et al.; Studies on the irreversible thermoinactivation of beta-amylase from Bacillus megaterium B6 exposed to 60 degrees C revealed that the deactivation mechanism probably results from the oxidation of thiols present at the active site of the enzyme . Several attempts were made to increase its thermostability, which indicated that Mn2+ played a key role in determining thermostability and partially reactivating the inactivated enzyme . Immobilization of beta-amylase through gel-entrapment and covalent crosslinking brought about a remarkable increase in thermotolerance with about a 14-fold increase in catalytic half-life.

Nature, 1994 Dec 8, 372(6506), 552 - 5
Systemic and mucosal immunity induced by BCG vector expressing outer-surface protein A of Borrelia burgdorferi; Langermann S et al.; The bacillus Calmette-Guerin (BCG) is a live attenuated strain of Mycobacterium bovis which offers potential advantages as a vector for mucosal delivery of antigens . Recombinant BCG elicits protective humoral immune responses to a variety of antigens . Furthermore, BCG binds specifically to microfold cells present in the epithelium overlying lymphoid follicles throughout the mucosal immune system . Here we show that a single intranasal vaccination with recombinant BCG expressing the outer-surface protein A antigen from B . burgdorferi results in a prolonged (more than one year) protective systemic IgG response and a highly sustained secretory IgA response which is disseminated throughout the mucosal immune system . Furthermore, intranasal immunization induces marked, organized lymphocyte accumulation in the proximal nasopharyngeal lymphoid tissue as well as at distal mucosal sites; the appearance and persistence of lymphoid aggregates correlates with the secretory immune responses . Thus intranasal immunization with recombinant BCG is a powerful method for inducing long-lasting secretory and systemic immune responses.

Gene, 1994 Dec 2, 150(1), 71 - 3
Sequence of the cloned bseCIM gene: M.BseCI reveals high homology to M.BanIII; Rina M et al.; The bseCIM gene, encoding M.BseCI methyltransferase (MTase) from a Bacillus stearothermophilus strain, has been previously cloned and expressed in Escherichia coli {Rina and Bouriotis, Gene 133 (1993) 91-94} . The nucleotide (nt) sequence of a 2357-bp BspMII-EcoRI fragment encoding bseCIM has now been determined . The sequence predicts a MTase of 579 amino acids (aa), 66.7 kDa . Comparison of the deduced aa sequence of M.BseCI with sequences of various MTases revealed a significant homology to m6A-MTases, especially to its isoschizomer M.BanIII from Bacillus aneurinolyticus.

Gene, 1994 Dec 2, 150(1), 203 - 4
Cloning and sequencing of the sspF (originally 0.3 kb) genes from Bacillus cereus and Bacillus megaterium; Loshon CA et al.; The sspF gene (originally 0.3 kb) of Bacillus cereus and B . megaterium has been cloned and sequenced, and the predicted amino acid sequences of the gene products (SspF) compared to that of B . subtilis SspF . These proteins exhibit an average of 74% sequence identity across species, suggesting they may play some important role in either sporulation or the dormant spore.

Eur J Biochem, 1994 Dec 1, 226(2), 277 - 83
Multiple proline substitutions cumulatively thermostabilize Bacillus cereus ATCC7064 oligo-1,6-glucosidase . Irrefragable proof supporting the proline rule; Watanabe K et al.; Nine residues of Bacillus cereus ATCC7064 oligo-1,6-glucosidase were replaced stepwise with proline residues . Of the nine residues, Lys121, Glu208 and Glu290 were at second sites of beta turns; Asn109, Glu175 and Thr261 were at N-caps of alpha helices; Glu216, Glu270 and Glu378 were in coils within loops . The replacements were carried out in the order, Lys121-->Pro, Glu175-->Pro, Glu290-->Pro, Glu208-->Pro, Glu270-->Pro, Glu378-->Pro, Thr261-->Pro, Glu216-->Pro and Asn109-->Pro . The resultant nine active mutant enzymes contained 1-9 more proline residues than B . cereus oligo-1,6-glucosidase . The thermostability of these mutants was additively enhanced with the increase in the number of proline residues introduced . The increase in the thermostability was most remarkable when proline residues were introduced at second sites of beta turns or at N-caps of alpha helices . The above results afforded irrefragable proof for the proline rule as an effective principle for increasing protein thermostability {Suzuki, Y., Oishi, K., Nakano, H . & Nagayama, T . (1987) Appl . Microbiol . Biotechnol . 26, 546-551}.

Cancer Immunol Immunother, 1994 Dec, 39(6), 401 - 6
Induction of TH1- and TH2-associated cytokine mRNA in mouse bladder following intravesical growth of the murine bladder tumor MB49 and BCG immunotherapy; McAveney KM et al.; Productive immunity to murine and human parasites is associated with the development of a type I T cell response (interferon-gamma-producing) while type II responses (interleukin-4-producing) suppress the development of delayed-type hypersensitivity (DTH) and the elimination of the parasite . To determine if a similar regulatory pathway might exist in tumor systems and may be effected by immunotherapeutic manipulation, we have studied the localized cytokine response to the murine bladder tumor MB49 growing intravesically in syngeneic mice . Intravesical growth of MB49 results in the host-derived expression of mRNA for both interleukin-4 (IL-4) (TH2) and interferon gamma (IFN gamma) (TH1), as well as tumor necrosis factor alpha (TNF alpha) expression of indeterminate origin . Intravesical instillation of bacillus Calmette-Guerin (BCG), highly effective in eliminating bladder tumors clinically and in experimental systems, results in IFN gamma and TNF alpha mRNa production in the bladder wall, but no IL-4 . Following BCG treatment of intravesical MB49, the number bladders expressing IL-4 mRNA decreases, while IFN gamma and TNF alpha expression remains constant . These results are consistent with the mechanism of action of BCG involving the generation of an enhanced TH1 immune milieu in the bladder wall, which may contribute to the generation of productive tumor-specific immunity.

Chest, 1994 Dec, 106(6 Suppl), 293S - 296S
Surgical adjuvant therapy for stage II and stage III adenocarcinoma and large cell undifferentiated carcinoma; Holmes EC; The Lung Cancer Study Group (LCSG) randomized 141 patients with resected stage II and III adenocarcinoma and large cell undifferentiated carcinoma to receive postoperative combined chemotherapy with cyclophosphamide, doxorubicin, and cisplatin (CAP) chemotherapy or bacillus Calmette-Guerin (BCG) and levamisole immunotherapy . Careful intraoperative staging was performed on all patients . Before randomization, patients were stratified by stage, weight loss, cardiac arrhythmia, and institution . Prognostic variables such as stage, age, weight loss, and nodal involvement were equally distributed between the two groups . Disease-free survival was significantly prolonged in the group receiving chemotherapy . There was no evidence of a deleterious effect of the immunotherapy . This study indicates that postoperative CAP chemotherapy is effective in prolonging disease-free survival in these patients.

Chest, 1994 Dec, 106(6 Suppl), 287S - 292S
A placebo-controlled randomized double-blind study of adjuvant intrapleural BCG in patients with resected T1N0, T1N1, or T2N0 squamous cell carcinoma, adenocarcinoma, or large cell carcinoma of the lung . LCSG Protocol 771; Gail MH; This article reviews the design and findings of LCSG Protocol 771, a randomized double-blind comparison of postoperative adjuvant intrapleural bacillus Calmette-Guerin (BCG) against saline solution placebo control in 473 patients with resected T1N0, T1N1, or T2N0 non-small cell lung cancer . Patients were randomized from August 30, 1977, through October 20, 1980, and follow-up ended on January 1, 1990 . There was no evidence of improved survival or time to recurrence among patients given BCG in contrast to earlier promising findings . A calculation suggests that false-positive results due to chance are not uncommon in small preliminary studies, indicating the need for larger confirmatory trials such as LCSG Protocol 771 . Other contributions from this Protocol are also reviewed, including data on BCG toxicity, immunologic effects, patterns of recurrence, and identification of prognostic risk groups.

J Bacteriol, 1994 Dec, 176(23), 7182 - 9
Comparative studies of S-layer proteins from Bacillus stearothermophilus strains expressed during growth in continuous culture under oxygen-limited and non-oxygen-limited conditions; Sara M et al.; The specific properties of S-layer proteins from three different Bacillus stearothermophilus strains revealing oblique, square, or hexagonal lattice symmetry were preserved during growth in continuous culture on complex medium only under oxygen-limited conditions in which glucose was used as the sole carbon source . When oxygen limitation was relieved, amino acids became metabolized, cell density increased, and different S-layer proteins from wild-type strains became rapidly replaced by a new common type of S-layer protein with an apparent subunit molecular weight of 97,000 which assembled into an identical oblique (p2) lattice type . During switching from wild-type strains to variants, patches of the S-layer lattices characteristics for wild-type strains, granular regions, and areas with oblique lattice symmetry could be observed on the surface of individual cells from all organisms . The granular regions apparently consisted of mixtures of the S-layer proteins from the wild-type strains and the newly synthesized p2 S-layer proteins from the variants . S-layer proteins from wild-type strains possessed identical N-terminal regions but led to quite different cleavage products upon peptide mapping, indicating that they are encoded by different genes . Chemical analysis including N-terminal sequencing and peptide mapping showed that the oblique S-layer lattices synthesized under increased oxygen supply were composed of identical protein species.

Infect Immun, 1994 Dec, 62(12), 5305 - 11
Non-major histocompatibility complex-restricted cytotoxic activity of blood mononuclear cells stimulated with secreted mycobacterial proteins and other mycobacterial antigens; Ravn P et al.; Several observations indicate that non-major histocompatibility complex (MHC)-restricted cytotoxicity, mediated for example by natural killer cells and lymphokine-activated killer cells, may serve as an important antimicrobial defense mechanism . The purpose of the present study was to investigate the influences of different mycobacterial antigens on non-MHC-restricted cytotoxicity and further to investigate the ways by which various lymphocyte subpopulations contribute to the development of this cytotoxicity . Non-MHC-restricted cytotoxicity was induced following stimulation of mononuclear cells with tuberculin purified protein derivative, Mycobacterium bovis bacillus Calmette-Guerin (BCG), short- and long-term culture filtrates of virulent Mycobacterium tuberculosis H37Rv, and 30-31-kDa secreted mycobacterial protein . These antigens also induced proliferation and production of gamma interferon . The CD4+ cells proliferated and expressed interleukin-2 receptors following stimulation with mycobacterial antigens . Depletion studies after antigen stimulation showed that the cytotoxic effector cells were CD16+ CD56+ and CD4-; the CD4+ cells alone did not mediate non-MHC-restricted cytotoxicity . To evaluate the influence of CD4+ cells on the development of non-MHC-restricted cytotoxicity, blood mononuclear cells were depleted of CD4+ cells before antigen stimulation . When mononuclear cells were incubated with purified protein derivative or short-term culture filtrate in the absence of CD4+ cells, cytotoxic activity was reduced . This reduction was abolished by interleukin-2 but not by gamma interferon . We conclude that several mycobacterial antigens are able to induce non-MHC-restricted cytotoxicity . This study indicates that non-MHC-restricted cytotoxicity following stimulation with mycobacterial antigens is induced by cytokines released by antigen-specific activated CD4+ cells.

Pediatr Dermatol, 1994 Dec, 11(4), 338 - 41
Bacillary angiomatosis in an immunocompetent child: the first reported case; Paul MA et al.; Bacillary angiomatosis, an infectious process associated with Rochalima spp., was thought until recently to be restricted to HIV-infected or otherwise immunosuppressed patients . In 1993, bacillary angiomatosis was reported in several immunocompetent adults . An extensive literature review failed to find references to bacillary angiomatosis in immunocompetent children . We describe a 6-year-old female who presented with a single, rapidly growing, friable, erythematous papule on her neck . Histologic examination of a biopsy specimen confirmed the diagnosis of bacillary angiomatosis . The patient was otherwise healthy, and her physical examination was normal . Laboratory studies, including HIV serology, were normal . The patient was treated with six weeks of oral erythromycin without evidence of recurrence . We present and discuss the implications of the first case of bacillary angiomatosis in an immunocompetent child.

Clin Infect Dis, 1994 Dec, 19(6), 1084 - 91
Serological analysis of pulmonary and extrapulmonary tuberculosis with enzyme-linked immunosorbent assays for anti-A60 immunoglobulins; Zou YL et al.; IgA, IgG, and IgM antibodies to mycobacterial antigen A60 were measured by ELISA in blood, pleural fluid, and cerebrospinal fluid from 560 patients with pulmonary and/or extrapulmonary tuberculosis who were being treated at hospitals in northern China and from 734 uninfected controls . Among 529 healthy persons (most of whom had been vaccinated with bacille Calmette-Guerin {BCG} and 287 of whom were tuberculin-positive), the rate of false-positive results was negligible; this observation ruled out interference of remote BCG vaccination with A60 assays at the chosen cutoff level . Rates of positivity for IgM and IgG, respectively, were 80% and 36% among patients with active primary pulmonary tuberculosis, 31% and 88.5% among patients with active postprimary pulmonary tuberculosis, 0 and 41% among patients with inactive pulmonary tuberculosis, and 30%-61% and 69%-86% among patients with extrapulmonary tuberculosis . Paired samples of blood and pleural fluid from patients with pleurisy contained IgA antibody to A60 at equal titers; in contrast, most patients with tuberculous meningitis (100% of whom had a positive ELISA result) had higher levels of IgG antibody to A60 in cerebrospinal fluid than in blood--proof of intrathecal synthesis.

Scand J Urol Nephrol, 1994 Dec, 28(4), 365 - 8
Bacillus Calmette-Guérin therapy for high-risk superficial bladder cancer; Thanos A et al.; Twelve consecutive patients with carcinoma in situ (CIS) and 17 with T1G3 transitional cell cancer of the bladder were given intravesical Bacillus Calmette-Guerin (BCG) therapy after transurethral resection of all visible lesions . BCG (150 mg in 50 ml normal saline) was given once weekly for 12 weeks and then once monthly for 6 months . The median follow-up was 2.5 years in the CIS group and 3 years in the T1G3 group, with cystoscopy, cytology and random biopsies performed every 3 months . Two of the 12 CIS tumours relapsed, with stage progression (T2G3) . Of the 17 T1G3 transitional cell cancers, two relapsed with Ta-1, G1-2 and four progressed to T2-3G3 requiring cystectomy (3) or irradiation (1) . One of these four patients died of metastatic disease . BCG therapy thus was effective in preventing progression CIS, but less useful for such purpose in T1G3 transitional cell tumours, for which it cannot be recommended as standard treatment.

Wei Sheng Wu Xue Bao, 1994 Dec, 34(6), 449 - 56
{Studies on the ecological distribution of Bacillus thuringiensis in forest soils of China}; Dai L et al.; A total of 384 forest soils samples at 0-5cm depth collected from 8 forest site zones (13 Natural Reserves) in China were examined for the distribution of Bacillus and Bacillus thuringiensis . The 79 cultures (4.21%) in 1873 isolates of Bacillus collected referable to the Bacillus thuringiensis . Soil pH, moisture content and soil nutrient were determined . Rules of the ecological distribution of Bacillus thuringiensis of 8 forest site zones in China were analysed . Toxicities against 6 species of insect were tested . Strain some efficient serains were obtained . It was shown that resource of Bacillus thuringiensis is much fruitful in forest soils of China.

Bull Pan Am Health Organ, 1994 Dec, 28(4), 312 - 23
HIV/AIDS and its interaction with tuberculosis in Latin America and the Caribbean; Zacarias F et al.; At present, human immunodeficiency virus (HIV) is thought to have infected over 17 million people worldwide, over 1 million in North America and roughly 2 million in Latin America and the Caribbean . By comparison, infection with the tuberculosis (TB) agent Mycobacterium tuberculosis is far more common, current estimates indicating that roughly one-third of the world's population is infected . These two infections tend to aggravate each other . That is, HIV leads to a progressive immune system depression that favors reactivation of TB in people with latent tuberculous infections; it promotes progression of TB primary infections or reinfections to full-blown tuberculous disease; and it fosters TB transmission, because those simultaneously infected with HIV and M . tuberculosis tend to develop a bacilliferous and contagious TB that can be transmitted to other susceptible individuals, even though the latter are HIV-negative . In addition, this coinfection tends to promote circulation of drug-resistant M . tuberculosis and to produce peculiar manifestations that complicate TB diagnosis, treatment, and control . Overall, it seems clear that the growing threat posed by these associated agents demands effective action in the form of well-coordinated measures involving throughgoing participation by all countries.

Protein Expr Purif, 1994 Dec, 5(6), 547 - 52
An improved system for ribonuclease Ba expression; Okorokov AL et al.; The extracellular ribonuclease from Bacillus amyloliquefaciens (barnase, RNase Ba) is a well-characterized enzyme extensively used in structure-function studies . A new system for efficient expression and purification of barnase has been developed . The strong regulated expression cassette with the Pr promoter of lambda phage and the cooperative expression of barnase and barstar under its control have been applied to expression of these proteins in Escherichia coli . The expression cassette containing the Pr promoter of E . coli lambda phage under cI repressor regulation and the nucleotide sequence coding for barnase and barstar structural genes were merged into the plasmid pTN441, which was used for large-scale barnase production . The phoA signal peptide was used to express the target protein into cell periplasm . The purification of RNase Ba was carried out in two steps: the initial sample was concentrated followed by RP-HPLC . The system provides a stable yield of homogeneous protein of about 100-150 mg per liter of culture medium.

Epidemiol Mikrobiol Imunol, 1994 Dec, 43(4), 158 - 61
{Drug resistance in M . tuberculosis in Prague 1987-1992}; Havelkova M et al.; In 1987-1992 in Prague drug resistance of M . tuberculosis to four standard antituberculotics (isoniazide, streptomycin, rifampicin, ethambutol) was recorded in 39 patients, i.e . in 1.5-5% of patients with bacillary tuberculosis . Initial resistance was found in 25, secondary resistance in 13 subjects, in one patient both types of resistance were observed . In both groups men and patients born before 1941 predominated . In patients with initial resistance findings rated as small or medium-sized predominated, while in secondary resistance half the findings were medium-sized or extensive; an extrapulmonary form was recorded in one female patient . In the group of patients with an initially resistant M . tuberculosis monoresistant strains predominated and the highest ratio was accounted for by strains resistant to isoniazide; in patients with secondary resistance strains with combined resistance to two or more drugs predominated, however in all instances a combination of isoniazide with other antituberculotics was involved . With regard to the changing epidemiological situation as regards tuberculosis (in particular the arrested decline of the incidence of bacillary tuberculosis and the increase of drug resistance of M . tuberculosis), the authors recommend further monitoring and epidemiological analysis of the incidence of strains with initial and secondary resistance, systematic external checks of laboratory technique used for testing the sensitivity to antituberculotics and its centralization as well as the introduction of analyses of restrictive fragments of the DNA genome (RFLP method) to monitor the spread of tuberculous mycobacteria in the population.

Gaoxiong Yi Xue Ke Xue Za Zhi, 1994 Dec, 10 Suppl, S102 - 8
Dengue vector control: present status and future prospects; Yap HH et al.; Dengue Fever (DF) and Dengue Haemorrhagic Fever (DHF) have been the most common urban diseases in Southeast Asia since the 1950s . More recently, the diseases have spread to Central and South America and are now considered as worldwide diseases . Both Aedes aegypti and Aedes albopictus are involved in the transmission of DF/DHF in Southeast Asian region . The paper discusses the present status and future prospects of Aedes control with reference to the Malaysian experience . Vector control approaches which include source reduction and environmental management, larviciding with the use of chemicals (synthetic insecticides and insect growth regulators and microbial insecticide), and adulticiding which include personal protection measures (household insecticide products and repellents) for long-term control and space spray (both thermal fogging and ultra low volume sprays) as short-term epidemic measures are discussed . The potential incorporation of IGRs and Bacillus thuringiensis-14 (Bti) as larvicides in addition to insecticides (temephos) is discussed . The advantages of using water-based spray over the oil-based (diesel) spray and the use of spray formulation which provide both larvicidal and adulticidal effects that would consequently have greater impact on the overall vector and disease control in DF/DHF are highlighted.

Appl Environ Microbiol, 1994 Dec, 60(12), 4614 - 6
Identification and analysis of the antigens detected by two commercial Bacillus cereus diarrheal enterotoxin immunoassay kits; Beecher DJ et al.; The usefulness of two commercial immunoassays for the detection of diarrheal enterotoxin of Bacillus cereus is unclear because the identity of the enterotoxin(s) has not been proven and the kits detect different proteins . We found that the Bacillus cereus Enterotoxin-Reversed Passive Latex Agglutination kit (Oxoid) detects the L2 component from hemolysin BL, and the Bacillus Diarrhoeal Enterotoxin Visual Immunoassay (Tecra) detects two apparently nontoxic proteins.

Appl Environ Microbiol, 1994 Dec, 60(12), 4596 - 9
Molecular analysis of mercury-resistant Bacillus isolates from sediment of Minamata Bay, Japan; Nakamura K et al.; Bacillus isolates from Minamata Bay sediment were selected for the ability to volatilize mercury from a range of organomercurials (including methylmercury) . Chromosomal DNA from 74 of 78 such strains hybridized with the mer mercury resistance operon DNA from marine Bacillus sp . strain RC607 (Y . Wang, M . Moore, H . S . Levinson, S . Silver, C . Walsh, and I . Mahler, J . Bacteriol . 171:83-92, 1989) . The most frequent classes with regard to restriction nuclease site maps of the mer operon for the new isolates were identical to or closely related to the mer determinant of strain RC607 . PCR amplification analysis with primers designed from the strain RC607 mer operon gave products of precisely the predicted size with the 74 Minamata Bay isolates.

Biosci Biotechnol Biochem, 1994 Dec, 58(12), 2283 - 5
Site-directed mutagenesis of the Asp-197 and Asp-202 residues in chitinase A1 of Bacillus circulans WL-12; Watanabe T et al.; The contribution of the Asp-197 and Asp-202 residues in chitinase A1 of Bacillus circulans WL-12 to the catalytic reaction was studied by site-directed mutagenesis of these residues . A kinetic analysis of the purified mutant chitinases suggests the involvement of both the Asp-197 and Asp-202 residues in the catalytic events of this enzyme, although the effects of mutations of Asp-197 were less severe than those of the other mutations.

Zentralbl Hyg Umweltmed, 1994 Dec, 196(4), 360 - 6
The effect of different water-insoluble anorganic salts on the resistance and storage time of Bacillus stearothermophilus spores used for biological indicators; Muller HE; The survival of Bacillus stearothermophilus spores was investigated over 7 years . The spores were precipitated with 18 insoluble inorganic salts . There were hydroxides, carbonates, oxalates, phosphates, and sulfate of aluminium, barium, calcium, iron, magnesium, manganese, and tin . The stability of the spores to dry and moist heat was strongest in precipitates of barium and calcium salts . Most phosphates and the heavy metals caused a rapid inactivation of spores . Finally, the survival of the spores in the 7 years of observation was the best in barium sulfate and calcium oxalate.

Mol Microbiol, 1994 Dec, 14(5), 939 - 46
Characterization of a gene responsible for the Na+/H+ antiporter system of alkalophilic Bacillus species strain C-125; Hamamoto T et al.; An alkali-sensitive mutant, 38154, of the alkalophilic Bacillus sp . strain C-125 could not grow at an alkaline pH . The nucleotide sequence of a 3.7 kb parental DNA fragment that recovers the growth of 38154 at alkaline pH has four open reading frames (ORF1-4) . By subcloning the fragment, we demonstrated that a 0.25 kb DNA region is responsible for the recovery . Direct sequencing of the mutant's corresponding region revealed a G to A substitution . The mutation resulted in an amino acid substitution from Gly-393 to Arg of the putative ORF1 product, which was deduced to be an 804-amino-acid polypeptide with a molecular weight of 89,070 . The N-terminal part of the putative ORF1 product showed amino acid similarity to those of the chain-5 products of eukaryotic NADH quinone oxidoreductases . Membrane vesicles prepared from 38154 did not show membrane potential (delta psi)-driven Na+/H+ antiporter activity . Antiporter activity was resumed by introducing a parental DNA fragment which recovered the mutant's alkalophily . These results indicate that the mutation in 38154 affects, either directly or indirectly, the electrogenic Na+/H+ antiporter activity . This is the first report which shows that a gene responsible for the Na+/H+ antiporter system is important in the alkalophily of alkalophilic microorganisms.

Mol Microbiol, 1994 Dec, 14(5), 851 - 60
Specificity domain localization of Bacillus thuringiensis insecticidal toxins is highly dependent on the bioassay system; Masson L et al.; The Bacillus thuringiensis crylA(a) and crylA(c) gene specificity regions were probed by creating and testing hybrid toxins both in vivo and in vitro against cultured insect cells or dissociated midgut epithelial cells . Toxin threshold dose determinations revealed that CrylA(c) is highly active against cultured Choristoneura fumiferana cells (CF-1) whereas CrylA(a) is nontoxic . In live insect bioassays, a reversed order of toxicity was observed . Hybrid analysis revealed that the CrylA(c) toxicity-determining region is located between codons 258 and 510 . Two smaller subsections of this region (residues 258-358 and 450-510) were able to confer toxicity, although at lower levels, and one region (358-450) was present where progressive substitutions of crylA(a) with crylA(c) sequences had no effect . Exchanging the non-homologous N-terminal regions of CrylA(c) with CrylE suggested that the N-terminus does not play a role in specificity . One hybrid clone, MP80, displays a 99.3% homology to CrylA(b) but shows an 800-fold increase in toxicity to CF-1 cells relative to that shown by CrylA(b) . Direct comparison between live Bombyx mori bioassays and a newly developed in vitro lawn assay using dissociated midgut epithelial cells from the same insect revealed striking differences in toxicity . The toxicity-determining region for B . mori larvae was determined to be between codons 283 and 450, although the 450-620 codon region may exert an influence on toxicity . In general, native or hybrid toxins showing little or no insect intoxication were very active against the epithelial cells, suggesting that factors other than toxin amino acid sequence play an important role in determining toxin specificity.

J Am Mosq Control Assoc, 1994 Dec, 10(4), 534 - 9
Malaria control utilizing Bacillus sphaericus against Anopheles stephensi in Panaji, Goa; Kumar A et al.; In a large malaria endemic area in Panaji city, Goa, India, the weekly application of the biolarvicide Bacillus sphaericus (Strain 101, Serotype H 5a 5b) at the rate of 1 g/m2 in the main Anopheles stephensi larval habitats, viz., curing waters, masonry tanks, and sump tanks (under construction), from April to December 1993 resulted in a sharp decline in the habitat positivity (range 0.13-8.0%) as compared with the rest of the Panaji (range 2.2-30.6%) where temephos (Abate) was used as the larvicide . Bacillus sphaericus spraying also led to a significant decline in anopheline densities in positive habitats (range 0-7.3/10 dips) as compared with control habitats (range 0.9-53.0/10 dips) . Concurrently, malaria incidence observed in the experimental area (slide positivity rate {SPR} range 2.3-7.8%; monthly parasite index {MPI} range 0.18-1.44) was lower than in the control area (SPR range 14.3-25.5%; MPI range 1.75-6.12).

J Am Mosq Control Assoc, 1994 Dec, 10(4), 527 - 33
Protocol for the introduction of new Bacillus thuringiensis Israelensis products into the routine mosquito control program in Germany; Becker N et al.; The efficacy of new and frequently used formulations of Bacillus thuringiensis israelensis (B.t.i.) has been evaluated in the laboratory and in the field conditions under which they will be used in Germany . The principles governing the successful introduction of new formulations of microbial control agents into routine programs have been highlighted . The potency of the formulations in use (Teknar, Bactimos, and Vectobac) and their efficacy against the indigenous mosquito species Aedes vexans, Aedes cantans, and Culex pipiens have been assessed to determine the minimum effective dosage in the laboratory and the optimum effective dosage in small field trials . These results should ensure the proper selection of the most appropriate formulation and dosage for the particular climatic and ecological conditions.

J Am Mosq Control Assoc, 1994 Dec, 10(4), 492 - 5
Evaluation of granular corncob formulations of Bacillus thuringiensis serovar israelensis against mosquito larvae using a semi-field bioassay method; Ali A et al.; A semi-field bioassay method using commonly available laundry tubs evaluated efficacy and residual activity of 6 granular corncob formulations of Bacillus thuringiensis serovar israelensis against laboratory-reared late 3rd- and early 4th-instar larvae of Aedes taeniorhynchus introduced to the tubs . These formulations produced appreciable larval mortalities (up to 75%) for 9 days posttreatment, with indications of having more prolonged activity . The consistent posttreatment larval mortality trends and the elucidated efficacy differences between formulations suggest that this bioassay method for the granular formulations is reliable and reproducible.

J Biochem (Tokyo), 1994 Dec, 116(6), 1202 - 4
Mutant genes of gramicidin S synthetase 1 defective in phenylalanine racemization have the same sequence as the wild gene; Hori K et al.; Mutant grs1 genes were cloned and sequenced from the Bacillus brevis Nagano BI-4, C-3, E-1, and E-2 strains, which produce defective gramicidin S synthetase 1 (GS1), lacking racemase activity . Surprisingly, these mutant genes had entirely the same sequence as that of the wild type gene . These mutant strains also produce defective gramicidin S synthetase 2 (GS2), lacking 4'-phosphopantetheine, a prosthetic group of this enzyme . The participation of this group in phenylalanine racemization is suggested.

Can J Microbiol, 1994 Dec, 40(12), 1026 - 34
Production of multiple delta-endotoxins by Bacillus thuringiensis: delta-endotoxins produced by strains of the subspecies galleriae and wuhanensis; Chestukhina GG et al.; A method was developed to assess the number of delta-endotoxins contained in Bacillus thuringiensis entomocidal crystals . It utilized proteolytic conversion of 130-kDa protoxin into 60- to 65-kDa "true" toxin via limited proteolysis with trypsin and separation of stable N-terminal domains by fast-performance liquid chromatography . Immunodiffusion experiments and N-terminal sequence determination (applied to the major component isolated by SDS-PAGE) completed the analysis of the crystal protein composition . The application of this approach to crystals produced by cells of B . thuringiensis subsp . galleriae and wuhanensis allowed us to identify at least seven and eight different delta-endotoxins, respectively . Among those delta-endotoxins assigned to previously described families, CryIA, CryID, CryIF, and CryIG were found, as well as crystal proteins, which possess N-terminal amino acid sequences very different from those of all known delta-endotoxins . Possible functional consequences of delta-endotoxin multiplicity are discussed.

Bioorg Khim, 1994 Dec, 20(12), 1334 - 41
{New site-specific endonucleases from strains of Bacillus}; Sokolov NN et al.; In a search for new restriction endonucleases type II, among forty bacterial strains of the Bacillus genus two strains producing site-specific endonucleases have been found . Endonucleases BbvAIII and BspFI, isolated from B . brevis BLM B-677 and B . species F, are shown to be true isoschisomers of BspMII (Kpn2I) and Sau3AI, respectively.

Bioorg Khim, 1994 Dec, 20(12), 1327 - 33
{Isolation, purification, and characteristics of the new restriction enzymes BciBI and BciBII, produced by Bacillus circulans}; Samko OT et al.; New site-specific endonucleases BciBI and BciBII have been detected in Bacillus circulans . The enzymes were purified by fractionation of cell-free extract with polyethylene imine and ammonium sulphate (40-80% of saturation) followed by chromatography on DEAE-sepharose, blue-sepharose and phosphocellulose . The endonucleases BciBI and BciBII were separated only at the final step of the purification--by chromatography on the phosphocellulose column . The yields of BciBI and BciBII were 600 and 10,000 U/g of cells . It was found that restriction endonucleases BciBI and BciBII are isoschizomers of ClaI and BstNI, respectively.

Am J Infect Control, 1994 Dec, 22(6), 329 - 33
Mycobacterium tuberculosis transmission rates in a sanatorium: implications for new preventive guidelines; Jernigan JA et al.; BACKGROUND: In 1990, the Centers for Disease Control and Prevention recommended substituting dust-mist particulate respirators for simple isolation masks in acid-fast bacillus isolation rooms, reasoning that air leaks around the simple masks could result in a higher rate of purified protein derivative skin-test conversion . In 1993, a Centers for Disease Control and Prevention draft guideline proposed that high-efficiency particulate air filter respirators be used instead of dust-mist particulate respirators . Epidemiologic data were not available to assess the importance of these changes or their cost-effectiveness . METHODS: The University of Virginia was affiliated with a tuberculosis hospital from 1979 until 1987 . We surveyed physicians who had served as residents in internal medicine during this period regarding purified protein derivative skin-test history . duration of work at the tuberculosis sanatorium, and any history of unprotected exposures to patients with active pulmonary or laryngeal tuberculosis . Patients with active tuberculosis at the sanatorium were isolated in negative-pressure rooms with UV lights . Physicians wore simple isolation masks in these rooms . RESULTS: Responses were received from 83 former resident physicians . Fifty-two physicians had worked on the tuberculosis wards for a total of 420 weeks, with no subsequent skin-test conversions (95% CI 0 to 1 conversion/8 physician-years) . CONCLUSIONS: These data document a low risk of occupational transmission of Mycobacterium tuberculosis to physicians who wear simple isolation masks in negative-pressure ventilation rooms with UV lights . This low rate predicts that the additional protective efficacy and cost-effectiveness of the more expensive high-efficiency particulate air filter respirators and the respiratory protection program will be low.

S Afr Med J, 1994 Dec, 84(12), 857 - 60
Fatal Capnocytophaga canimorsus (DF-2) septicaemia . A case report; Linton DM et al.; A 45-year-old man died 2 months after being bitten on the hand by a dog . He developed the rare but characteristic clinical picture of fulminant septicaemia and peripheral gangrene caused by a Gram-negative bacillus, Capnocytophaga canimorsus, previously known as dysgonic fermenter type 2 (DF-2), which is an occasional commensal in the oral flora of dogs and cats . This disease must be anticipated and dog bites appropriately managed to avoid the mortality associated with infection by this micro-organism . Initial treatment includes appropriate prophylactic antibiotics and debridement, while early exchange transfusion and emergency amputation may be of value in fulminant cases.

Biosci Biotechnol Biochem, 1994 Dec, 58(12), 2275 - 7
Construction of a new teichuronopeptide-defective derivative from alkaliphilic Bacillus sp . C-125 by cell fusion; Ito M et al.; Cell walls of facultative alkaliphilic Bacillus sp . C-125 are composed of peptidoglycan, teichuronic acid, and teichuronopeptide . A mutant lacking teichuronic acid, or defective in both teichuronic acid and teichuronopeptide has been isolated from the organism . We now constructed another type of a cell-wall defective mutant that was defective in teichuronopeptide but had teichuronic acid, by cell fusion using protoplasts prepared from a wild-type strain and a mutant defective in teichuronopeptide and teichuronic acid . This mutant grew more poorly than wild type or a teichuronic acid-defective strain of C-125 . The growth, however, was faster than that of the parental strain defective in both teichuronic acid and teichuronopeptide.

Virology, 1994 Dec, 205(2), 430 - 7
Detection of rice tungro bacilliform virus gene products in vivo; Hay J et al.; To study the products of the open reading frames (ORFs) of rice tungro bacilliform virus in rice plants the sequences containing ORFs I (encoding a 24-kDa protein, P24) and IV (P46) and the protease and polymerase (reverse transcriptase+RNaseH) domains of ORF III were cloned into a pGEX expression vector . The proteins, which were C-terminal fusions to glutathione S-transferase, were expressed in Escherichia coli and antisera were raised against them which, together with an antiserum against virus particles, was used to probe blots of proteins from infected and uninoculated plants and from virus preparations . The P24 antiserum detected virus-specific proteins of 74, 60, and 52 kDa, which are much bigger than expected . These proteins were found in virus preparations and immunogold labeling suggested that they might be internal in the particles . Virus-specific proteins of 33, 37, 62, and > 150 kDa were revealed by antiserum to virus particles . The antiserum to the protease revealed proteins of 13.5, 37, and 68 kDa both in extracts from infected plants and in purified virus preparations . This antiserum decorated intact virus particles as did the particle antiserum . The polymerase domain antiserum reacted with products of 56, 65, and 68 kDa in extracts from infected plants but not in virus particles . The antiserum to the ORF IV product did not detect any bands in either infected plant extracts or virus preparations . The significance of these products is discussed.

Infect Immun, 1994 Dec, 62(12), 5319 - 26
T-cell determinants and antibody binding sites on the major mycobacterial secretory protein MPB59 of Mycobacterium bovis; Roche PW et al.; Among the first proteins encountered by the host immune system upon infection or vaccination with mycobacteria are those secreted by the bacillus during growth . The antigen 85 complex of Mycobacterium bovis bacillus Calmette-Guerin (BCG) is composed of three closely related members . The mature 85B protein of M . bovis (MPB59) has a high degree of amino acid identity with the M . bovis 85A protein (76%) and the Mycobacterium tuberculosis 85B (99%) and 85A (76%) proteins . We have examined the regions of MPB59 which stimulate human T- and B-cell responses by use of a set of 28 synthetic peptides, 20 amino acids (aa) in length and overlapping by 10 aa . Initial proliferative assays with recombinant MPB59 demonstrated that peripheral blood mononuclear cells from 95% of BCG vaccinees and 52% of tuberculosis patients responded to the whole mature protein . Peripheral blood mononuclear cells from MPB59 responders, but not nonresponders, were stimulated by peptides in a dose-dependent fashion . Five peptides were reactive in more than half of the MPB59 responders . The T-cell-reactive regions were essentially identical in the M . bovis and M . tuberculosis 85B proteins . Subjects with a variety of HLA-DR phenotypes responded to a number of these peptides . The dominant T-cell-reactive regions were distinct from the peptides recognized by sera from tuberculosis patients (aa 71 to 100) and the murine monoclonal antibody HYT27 (aa 61 to 90) . The region reactive with antibodies overlapped part of the MPB59 sequence recently shown to participate in the binding of MPB59 to fibronectin.

Biochem Biophys Res Commun, 1994 Nov 30, 205(1), 113 - 9
Differential hydrolysis of immobilized phosphatidylcholines by phospholipases A2 and C; Delfino JM et al.; A novel phospholipid, 1-fatty acyl-2-(12-aminododecyl) phosphatidylcholine (APC), was synthesized and reacted with two different activated agarose matrices, differing in the spacer arm length: N-hydroxysuccinimidylester agarose (1-atom spacer arm) and N-hydroxysuccinimidylester-6-aminohexanoic acid agarose (8-atom spacer arm) . Both immobilized phosphatidylcholines were readily degraded by Bacillus cereus phospholipase C at similar rates . By contrast, Crotalus adamanteus phospholipase A2 hydrolyzed long-spacer arm phosphatidylcholine, but had less than one tenth of the activity towards the short-spacer arm one . These results are interpreted in terms of a chain length-related steric hindrance caused by the matrix, affecting phospholipase A2 but not phospholipase C activity, supporting the view that the first involves a deeper burrowing of the substrate into the enzyme molecule.

FEBS Lett, 1994 Nov 14, 354(3), 305 - 6
Mutational analysis of the active site of RNase of Bacillus intermedius (BINASE); Yakovlev GI et al.; To elucidate the functional role of some residues in the active site of binase, the extracellular ribonuclease of Bacillus intermedius, we used site-directed mutagenesis . On cleavage of various substrates the catalytic activity of binase mutant His101 Glu is 2.0-2.7% of that for the native enzyme . The decrease in activity is determined mainly by the decrease in molecular rate constant kcat, with almost unchanged affinity of the enzyme for the substrate, characterized by KM . This is the expected result if His101 acts as an general acid, donating a proton to the leaving group on cleavage of a phosphodiester bond . The replacement of Lys26 by Ala causes a reduction in the enzyme activity to 13-33%, depending on the substrate . The activity decreases are due to changes in both kcat and KM for poly(A) and poly(A) but in kcat alone for GpA . In the latter case the effect is far less than that seen in the homologous mutation in the closely related enzyme, barnase.

Biochim Biophys Acta, 1994 Nov 11, 1201(2), 245 - 50
The interaction of cytochrome c and the heme domain of cytochrome P-450BM-3 with the reductase domain of cytochrome P-450BM-3; Klein ML et al.; Cytochrome P-450BM-3 from Bacillus megaterium is a soluble, catalytically self-sufficient fatty acid mono-oxygenase that resembles the Class II P-450 systems of the eukaryotic endoplasmic reticulum . Its single polypeptide chain contains both a P-450 heme domain and an NADPH:P-450 reductase domain, each of which bears significant structural and functional homology with its microsomal counterparts . We report here that cytochrome c, which can accept NADPH-derived electrons from the reductase domain of P-450-BM-3, did not inhibit myristate hydroxylation catalyzed by P-450BM-3 or by two reductase domain mutant enzymes (W574Y, W574F) which have diminished hydroxylase activity relative to wild-type enzyme but retain cytochrome c reductase activity levels comparable to wild-type enzyme . Because reduced cytochrome c generated independently of the reductase domain of P-450BM-3 did not support myristate hydroxylation, it seems likely that cytochrome c binds to a site on the reductase domain which does not overlap the site of the heme domain interaction . We also found that myristate did not inhibit P-450BM-3-mediated cytochrome c reduction . Since neither substrate inhibited the conversion of the other, we conclude that the rate-limiting steps for both myristate hydroxylation and cytochrome c reduction by P-450BM-3 do not involve electron transfer through the reductase domain.

Biochim Biophys Acta, 1994 Nov 10, 1224(2), 247 - 54
Hydrolysis of cell surface inositol phospholipid leads to the delayed stimulation of phosphatidylinositol synthesis in bovine aortic endothelial cells; Sillence DJ et al.; In order to address the issue of how inositol phospholipid synthesis is controlled in a resting cell we looked for enhanced {3H}phosphatidylinositol (PtdIns) labelling in response to the hydrolysis of cell surface PtdIns . Bacillus thuringiensis PtdIns-PLC when added to intact bovine aortic endothelial (BAE) cells rapidly hydrolysed 9.1 +/- 1% of the total cellular PtdIns . This result suggests that BAE cells have a cell surface pool of PTdIns . Hydrolysis of cell surface PtdIns, in contrast to the agonist-stimulated hydrolysis of inner leaflet PtdIns, did not lead to a rapid (minutes) stimulation of PtdIns resynthesis . Prolonged incubation of BAE cells with PtdIns-PLC led to further hydrolysis of PtdIns (up to 20% of total cellular PtdIns) . This second phase of PtdIns-PLC induced hydrolysis was inhibited by the addition of brefeldin A suggesting that it was dependent on vesicular traffic to the plasma membrane from the endoplasmic reticulum . Furthermore, the above result suggests that prolonged incubation of intact cells with PtdIns-PLC leads to the slow depeletion of intracellular PtdIns stores . This second phase of PtdIns-PLC induced hydrolysis was associated with PtdIns resynthesis since prolonged incubation with PtdIns-PLC, but not B . cereus PtdCho-PLC (which does not hydrolyse PtdIns), led to enhanced PtdIns labelling . The results indicate that extracellular PtdIns-PLC induced PtdIns resynthesis may occur due to PtdIns-PLC induced intracellular PtdIns depletion.

Acta Eur Fertil, 1994 Nov-Dec, 25(6), 343 - 6
Puberal hormonal status and vaginal microbiology; Tolino A et al.; The Autors have examinated endocrine function in girls during puberal age . Have been showed datums relative to cultures effected on vaginal secrets withdrawn from little girls in prepuberal age . They have also studied polimicrobic vaginal flora related to sexual hormone levels . Have been controllated correlations between vaginal microbic status and cytohormonal response at different hormonal situation . The Doderlein presence is as much greater so higher is the oestrogen and progesterone (E/P) ratio, while the Doderlein bacillus isn't present when E/P ratio is lower . Perhaps in puberal age other factors than oestrogen and progesterone levels, could influence vaginal polymicrobic flora.

Appl Environ Microbiol, 1994 Nov, 60(11), 4199 - 202
Expression of mosquitocidal toxin genes in a gas-vacuolated strain of Ancylobacter aquaticus; Yap WH et al.; A series of plasmids bearing the binary toxin genes of Bacillus sphaericus 2297 or 2317.3, the 100-kDa toxin gene of B . sphaericus SSII-1, or the 130-kDa (cryIVB) toxin gene of Bacillus thuringiensis subsp . israelensis were constructed and introduced into Ancylobacter aquaticus by electroporation . The transformed A . aquaticus cells exhibited significant toxicity towards mosquito larvae, demonstrating a potential use of recombinant A . aquaticus for biological control of mosquitoes.

J Infect Dis, 1994 Nov, 170(5), 1326 - 30
Differential T cell responses to mycobacteria-secreted proteins distinguish vaccination with bacille Calmette-Guérin from infection with Mycobacterium tuberculosis; Roche PW et al.; The immune responses of healthy recipients of Mycobacterium bovis bacille Calmette-Guerin (BCG) vaccine, tuberculosis (TB) patients, and contacts of TB patients were examined to three major secretory proteins of Mycobacterium tuberculosis, MPB59, MPB64, and MPB70 . MPB59 evoked a T cell response in 78% of BCG vaccines, 62% of TB patients, and 60% of contacts . MPB64 and MPB70 were recognized by < 15% of BCG vaccinees, half of TB patients, and three-quarters of contacts . TB and leprosy patients had antibody responses to MPB59, but few had antibodies to MPB64 or MPB70 . Hybridization of mycobacterial DNA with specific gene probes demonstrated the absence of a gene for MBP64 in the vaccine strain of BCG, but the MPB70 gene was found in all virulent and vaccine BCG strains tested . Since MPB64 and MPB70 can induce delayed-type hypersensitivity reactions in infected animals, either of these proteins may have potential as skin test reagents for detecting infection with M . tuberculosis.

J Bacteriol, 1994 Nov, 176(22), 7032 - 7
Autoprocessing of the protease that degrades small, acid-soluble proteins of spores of Bacillus species is triggered by low pH, dehydration, and dipicolinic acid; Illades-Aguiar B et al.; The sequence-specific protease (termed GPR) that degrades small, acid-soluble proteins (SASP) during germination of spores of Bacillus species is synthesized during sporulation as an inactive precursor termed P46 . Approximately 2 h later in sporulation, P46 is converted proteolytically to a smaller form, termed P41, which is active in vitro, but which does not act significantly on SASP in vivo until spore germination is initiated . While it appears likely that P46-->P41 conversion is an autoprocessing event, the mechanisms regulating P46-->P41 conversion in vivo are not clear . In this work we found that P46-->P41 conversion in vitro was stimulated tremendously in an allosteric manner by pyridine-2,6-dicarboxylic acid (dipicolinic acid {DPA}) plus Ca2+ but not by Ca2+ in combination with a variety of DPA analogs . The processing reaction stimulated by Ca(2+)-DPA was seen at pH 5.1 but not at pH 6.2 or 7, and under these conditions P46-->P41 conversion exhibited a linear time course and was a first-order reaction, indicative of an intramolecular autoprocessing reaction . At pH 5.1, P46-->P41 conversion was stimulated markedly by very high ionic strength . At pHs from 5.1 to 6.6, P46-->P41 conversion also occurred when P46 was dehydrated to approximately 54% relative humidity . This processing was stimulated markedly when dehydration was carried out in the presence of DPA and NaCl but was greatly decreased when dehydration was to 10, 33, or 75% relative humidity . Since previous work has shown that P(46)-->P(41) processing in vivo takes place (i) after a fall in forespore pH to 6.3 to 6.9 and approximately in parallel with (ii) DPA accumulation by the forespore and (iii) dehydration of the forespore, out new finings in vitro suggest that these three changes may synergistically trigger P(46)-->P(41) autoprocessing in the developing forespore . Presumably the conditions in vivo during this authoprocessing preclude significant attack of the P(41) generated on its SASP substrates.

J Bacteriol, 1994 Nov, 176(21), 6464 - 9
Properties of two different Na+/H+ antiport systems in alkaliphilic Bacillus sp . strain C-125; Kitada M et al.; Na+/H+ antiport was studied in alkaliphilic Bacillus sp . strain C-125, its alkali-sensitive mutant 38154, and a transformant (pALK2) with recovered alkaliphily . The transformed was able to maintain an intracellular pH (pHin) that was lower than that of external milieu and contained an electrogenic Na+/H+ antiporter driven only by delta psi (membrane potential, interior negative) . The activity of this delta psi-dependent Na+/H+ antiporter was highly dependent on pHin, increasing with increasing pHin, and was found only in cells grown at alkaline pH . On the other hand, the alkali-sensitive mutant, which had lost the ability to grow above pH 9.5, lacked the delta psi-dependent Na+/H+ antiporter and showed defective regulation of pHin at the alkaline pH range . However, this mutant, like the parent strain, still required sodium ions for growth and for an amino acid transport system . Moreover, another Na+/H+ antiporter, driven by the imposed delta pH (pHin > extracellular pHout), was active in this mutant strain, showing that the previously reported delta pH-dependent antiport activity is probably separate from delta psi-dependent antiporter activity . The delta pH-dependent Na+/H+ antiporter was found in cells grown at either pH 7 or pH 9 . This latter antiporter was reconstituted into liposomes by using a dilution method . When a transmembrane pH gradient was applied, downhill sodium efflux was accelerated, showing that the antiporter can be reconstituted into liposomes and still retain its activity.

Chest, 1994 Nov, 106(5), 1624 - 6
Granulomatous pneumonitis following intravesical BCG . What therapy is needed?
LeMense GP, Strange C.
A 68-year-old man developed fever, cough, and dyspnea after intravesical bacillus Calmette-Guerin (BCG) . Chest radiograph revealed diffuse reticulonodular infiltrates with caseating granulomas on transbronchial biopsy specimen . Cultures were negative and the patient's condition improved with corticosteroids . The mechanism for BCG-induced granulomatous inflammation is poorly understood . Optimal therapy includes corticosteroids.

Clin Exp Immunol, 1994 Nov, 98(2), 229 - 33
Heat shock proteins as carrier molecules: in vivo helper effect mediated by Escherichia coli GroEL and DnaK proteins requires cross-linking with antigen; Barrios C et al.; In the past few years we have shown that mycobacterial heat shock proteins (hsp) of 65 and 70 kD exert a very strong helper effect in mice and monkeys when conjugated to peptides and oligosaccharides and given in the absence of adjuvants . In the present study we show that this adjuvant-free helper effect (i) is not due to lipopolysaccharide (LPS), since it was observed in LPS-resistant mice (C3H/HeJ) immunized with hsp-based constructs containing the malaria peptide (NANP)40, and (ii) is characteristic of hsp, since it was not observed with conjugates containing the mycobacterial p38 antigen, which is not a stress protein . Interestingly, the hsp GroEL and DnaK of Escherichia coli, which share a high degree of homology with the mycobacterial 65-kD and 70-kD hsp, respectively, exhibit a strong in vivo helper effect when conjugated to the (NANP)40 peptide, and the conjugates given in the absence of adjuvants . This in vivo helper behaviour of the GroEL and DnaK proteins corresponds well to that observed with the mycobacterial 65-kD and 70-kD hsp, respectively, since the hsp65- and GroEL-based constructs require previous priming of the animals with live bacille Calmette-Guerin (BCG), which is not needed for the hsp70- and DnaK-based constructs . Finally, using both mycobacterial and E . coli hsp we show that their in vivo helper effect in the absence of adjuvants requires cross-linking to the synthetic peptide . Taken together, our results suggest that the adjuvant-free helper effect observed with mycobacterial and E . coli hsp may be a generalized phenomenon, exhibited by hsp from diverse microorganisms . These findings may find applications in the design of vaccine constructs.

Virology, 1994 Nov 1, 204(2), 626 - 33
Replication intermediates of rice tungro bacilliform virus DNA support a replication mechanism involving reverse transcription; Bao Y et al.; Rice tungro bacilliform virus (RTBV) replication intermediates have been studied in rice plants infected with the virus . Unencapsidated virus-specific molecules were identified which had open circular, linear, supercoiled (SC), strong-stop, single-stranded, linear double-stranded hairpin, and double-stranded with single-stranded extension DNA forms . The structures of these different DNA forms were consistent with the replication model of cauliflower mosaic virus and support other results that reverse transcription is involved in the replication of RTBV . The existence of nonspecific and defective (+)-strand priming is suggested . The relative amount of SC DNAs differs in various tissues of the same plant and in the same tissue at different ages . This indicates host regulation of the virus replication cycle and a feedback regulatory mechanism in controlling the SC DNA level . There are no obvious differences in the composition of the replication intermediates between insect-infected and agroinoculated rice plants.

J Urol, 1994 Nov, 152(5 Pt 1), 1424 - 8
Intravesical bacillus Calmette-Guerin for superficial bladder cancer: experience with Danish 1331 strain; Kamat MR et al.; A total of 95 patients with stage Ta/T1 superficial bladder cancer was eligible for intravesical bacillus Calmette-Guerin (BCG) immunoprophylaxis according to the standard high risk criteria for tumor recurrence and progression . Of these patients 50 agreed to undergo treatment while the remaining 45 refused any intravesical therapy and served as concurrent nonrandomized controls . The patient and tumor characteristics in the 2 groups of patients were identical . All patients underwent complete transurethral resection of the bladder tumor . Therapy consisted of 120 mg . weekly instillations of BCG (Danish 1331 strain) for 6 weeks . Among the 50 patients 33 received BCG initially while 17 received it after failure of intravesical chemotherapy . Responses were evaluated according to standard criteria . Recurrence was noted in 24 patients (48%) in the BCG group compared to 38 (84.4%) in the control group . The recurrence rates per year and per 100 patient-months were significantly reduced in the BCG arm compared to the control arm . The mean interval to first recurrence and the mean recurrence interval were significantly increased in the BCG arm compared to the control arm . The relative risk of recurrence in the BCG group was 0.62 versus 1.63 in the control group . Subgroup analysis showed significant benefit of BCG for patients with single as well as multiple, stages Ta and T1, and grades II and III tumors . Comparison with pretreatment controls in the BCG group revealed a significant reduction in the recurrence rate in those patients after treatment with BCG, which was not seen in the control group . The benefit of BCG was seen in those who received BCG initially as well as in those who received it after failure of intravesical chemotherapy . Multivariate analysis of prognostic factors showed that this benefit in the BCG group was related only to the treatment, while other prognostic parameters, such as tumor stage, grade, number and so forth were not independent prognostic variables . The rate of progression to muscle invasion was not significantly different in the 2 treatment groups . The relapse-free survival in the BCG group was 35.4% at 60 months compared to 11.2% in the control group (p < 0.001) . The side effects of BCG therapy were mild, brief and easily controlled with conservative measures.

J Pharm Pharmacol, 1994 Nov, 46(11), 863 - 6
In-vivo and in-vitro targeting of a murine sarcoma by gelatin microparticles loaded with a glycan (PS1); Lou Y et al.; PS1, a complex polysaccharide derived from Mycobacterium bovis (Bacillus Calmette-Guerin, BCG) with considerable antitumor activity in-vivo, was loaded onto gelatin microparticles (mean diam . 1.45 micron) at a level shown to not produce the burst effect often seen with drug-loaded microparticulate systems . In-vitro dissolution experiments had demonstrated a sustained-release behaviour, with a half-life of approximately 8 h for what is an extremely water-soluble material . These PS1/gelatin systems had no measurable cytotoxicity against an S180 murine sarcoma cell in-vitro although fibronectin-mediated targeting of the microparticles for the tumour cells could be demonstrated . Injection into mice, with the S180 cells, of PS1 solutions or suspensions of PS1-loaded gelatin microparticles resulted in almost identical dose-related suppression for the tumour cell growth . When injected at intervals following injection of the tumour cells, however, for a period of 24-48 h there was a relatively enhanced activity of the formulated PS1, compared with the aqueous solution, after which both formulated and unformulated material became progressively less effective.

J Biochem (Tokyo), 1994 Nov, 116(5), 943 - 5
Prolyl aminopeptidase is not a sulfhydryl enzyme: identification of the active serine residue by site-directed mutagenesis; Kitazono A et al.; Prolyl aminopeptidase (PAP) has been classified as a sulfhydryl enzyme on the basis of its high sensitivity to p-chloromercuribenzoate and heavy metals . Recently, however, the possibility of PAP being instead a serine enzyme has been raised as a result of two observations--the conservation of some residues among the PAPs hitherto sequenced, and a similarity to some serine hydrolases . This is the first report describing an attempt to identify the active residue by site-directed mutagenesis . The pap genes from Bacillus coagulans and Aeromonas sobria, were used for the study . The changes made were Cys62Ser and Ser101Ala for the first enzyme, and Thr92Ala and Ser146Ala for the second . For both enzymes, only the changes made on the serine residues resulted in their complete inactivation, indicating that PAP is a serine peptidase.

Food Addit Contam, 1994 Nov-Dec, 11(6), 641 - 7
Oxytetracycline residue levels in chicken eggs after oral administration of medicated drinking water to laying chickens; Omija B et al.; Twenty laying birds were divided into four groups (n = 5) and given drinking water containing 0, 400, 600, and 800 mg/l of oxytetracycline respectively for 7 days . Eggs were collected continuously for 17 days after drug administration and stored at +4 degrees C . The oxytetracycline residues in yolk and albumen were analysed using a microbiological method with Bacillus cereus var . mycoides ATCC 11778 as the test organism . The mean maximum concentration of oxytetracycline was observed 2 days earlier in the albumen than in the yolk . The mean values in yolk and albumen were 0.526 and 0.280 mg/kg respectively . The depletion period was shorter for albumen than for yolk and oxytetracycline was detected in the yolk and albumen up to days 13 and 10 respectively . Withdrawal periods depended on the concentration of the antibiotic administered . Oxytetracycline residues reached a peak faster in albumen than in yolk, although the residues persisted for longer periods in the yolk.

Vet Microbiol, 1994 Nov, 42(2-3), 135 - 45
Tuberculosis in imported hyrax (Procavia capensis) caused by an unusual variant belonging to the Mycobacterium tuberculosis complex; Cousins DV et al.; Tuberculosis was diagnosed in an adult female hyrax (Procavia capensis) imported from South Africa and held in a captive colony at the Perth Zoo . An organism similar to Mycobacterium microti was isolated from the lung of this animal and the lung of an adult male hyrax in the colony . The organism was not pathogenic to rabbits or guinea pigs . Protein profiles and RFLP patterns using the probes IS6110 and pTBN12 showed both hyrax isolates were identical . These isolates were similar to a M . tuberculosis complex strain isolated from dassies (hyrax) in the late 1950s in South Africa and to M . microti, but appeared to be more closely related to the "dassie bacillus" . It is likely that at least one of the hyrax was infected at the time of collection in South Africa . The finding of tuberculosis in these imported animals highlights concern over the lack of suitable tests for the detection of tuberculosis in unusual animal species such as these, and the problems that can arise as a result of the importation of infected animalsPublication Types:

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