J Eukaryot Microbiol, 1993 May-Jun, 40(3), 336 - 40 Lipophosphoglycan antigen shedding by Leishmania donovani; Kaneshiro ES et al.; The biochemical characterizations of lipophosphoglycans from various Leishmania species reported by other workers may or may not contain several types of lipophosphoglycan molecules . This is the first report in which a specific lipophosphoglycan has been defined by both its antigenic and electrophoretic properties . Furthermore, a purification procedure for this specific lipophosphoglycan is described and some biochemical characterizations are presented . Phospholipase C and the so-called phosphatidylinositol-specific phospholipase C of Bacillus cereus convert the amphipathic form of the lipophosphoglycan antigen to the hydrophilic form . Under equivalent incubation conditions, other phospholipases tested were not effective in conversion of the amphipathic to the hydrophilic form . Since the amphipathic form is present in conditioned media, antigen shedding cannot be explained by phospholipase C digestion of the amphipathic form, which would result in the release of only the hydrophilic form into the medium . Both the pellet and the supernatant fractions of conditioned media contained both forms of the antigen and did not differ in the relative amounts of the two . This observation rules out membrane blebbing as the major mechanism for the release of the amphipathic form. J Eukaryot Microbiol, 1993 May-Jun, 40(3), 323 - 8 Antagonistic action of the bacterium Bacillus licheniformis M-4 toward the amoeba Naegleria fowleri; Cordovilla P et al.; Free-living amoebae belonging to the species Naegleria fowleri are known to be the etiological agents for a form of fulminant meningoencephalitis that is generally fatal (primary amoebic meningoencephalitis) . In a broad bacterial screening from soil and water we have isolated three strains (M-4, D-13 and A-12) belonging to the species Bacillus licheniformis that have remarkable amoebicidal activity against Naegleria sp . and also against different Gram-positive and Gram-negative bacteria . Physical-chemical characteristics, partial purification and biological activities of a substance produced by the M-4 strain have been investigated . This substance (m-4) is stable at high temperature (up to 100 degrees C) and extremes of pH (2.5-9.5) and also at -20 degrees C for months . Its production is greatly influenced by oxygenation of the cultures and is probably related to the sporulation process of the bacterium . Scanning electron microscope observations reveal that amoebae are lysed after a few minutes contact with m-4. Plant Mol Biol, 1993 May, 22(2), 313 - 21 Genetically improved potatoes: protection from damage by Colorado potato beetles; Perlak FJ et al.; Russet Burbank potato plants have been genetically improved to resist insect attack and damage by Colorado potato beetles (Leptinotarsa decemlineata (Say)) by the insertion of a cryIIIA gene encoding the insect control protein of Bacillus thuringiensis var . tenebrionis . A modified gene that dramatically improved plant expression of this protein was utilized . Its expression in Russet Burbank potato plants resulted in protection from damage by all insect stages in the laboratory and in dramatic levels of protection at multiple field locations . Analysis of these genetically modified potatoes indicated that they conform to the standards for Russet Burbank potatoes in terms of agronomic and quality characteristics including taste. J Clin Microbiol, 1993 May, 31(5), 1136 - 42 Intracellular bacteria of porcine proliferative enteropathy: cultivation and maintenance in vitro; Lawson GH et al.; An obligate intracellular bacterium was isolated from the intestines of all 10 cases of porcine proliferative enteropathy from four different pig farms . The organism grew in a rat enterocyte cell line (IEC-18) and was maintained over 20 passages . The growth of the bacteria was assessed by immunostaining of cells exposed to infection . Infection was not associated with morphological cell change, and growth was confined to cells infected at the time of each transfer of infection and the progeny of these cells . The bacterium is a microaerophilic, cell dependent, curved or rod-shaped, gram-negative bacillus that multiplies freely in the enterocyte cytoplasm . Cell cultures containing the intracellular bacteria appear to be free of other microorganisms, including chlamydiae and viruses. J Pediatr, 1993 May, 122(5 Pt 1), 697 - 702 Safety and immunogenicity of bacille Calmette-Guérin, diphtheria-tetanus-pertussis, and oral polio vaccines in newborn children in Zaire infected with human immunodeficiency virus type 1; Ryder RW et al.; OBJECTIVE: To determine the safety and immunogenicity of childhood vaccines in children with perinatally acquired human immunodeficiency virus type 1 (HIV-1) infection . DESIGN: Nonrandomized, prospective cohort study; 12-month follow-up period . SETTING: Obstetric wards and outpatient pediatric clinics at two large hospitals in Kinshasa, Zaire . PATIENTS: A total of 8108 pregnant women were screened for HIV-1 antibodies . The 474 children born to 466 seropositive women identified during screening and the 616 children born to 606 seronegative, age- and parity-matched women were vaccinated . INTERVENTION: The following vaccines were administered at the stated ages: bacille Calmette-Guerin (BCG) vaccine (2 days); trivalent oral Sabin poliomyelitis vaccine (2 days and 6, 10, and 14 weeks); and adsorbed diphtheria-tetanus-pertussis (DTP) vaccine (6, 10, and 14 weeks) . MEASUREMENTS AND MAIN RESULTS: Protective antibody titers to tetanus and poliovirus types 1, 2, and 3 were achieved in 95% of all children . Among children with HIV-1 infection, 70.8% had protective antibody titers to diphtheria compared with 98.5% of uninfected children (p < 0.05) . Geometric mean antibody titers to diphtheria and poliovirus types 1, 2, and 3 were significantly lower in children with HIV-1 infection than in uninfected children . Vaccine-associated side effects were similarly low in all children . CONCLUSIONS: The low incidence of side effects and the high proportion of children with HIV-1 infection who achieved protective postimmunization antibody titers support the continuing use of BCG, DTP, and oral polio vaccines in childhood immunization programs in HIV-1 endemic areas. Curr Opin Oncol, 1993 May, 5(3), 574 - 80 Advances in bladder cancer; Ozen H; Intravesical chemotherapy and immunotherapy are administered to patients with bladder cancer in the hope of decreasing the rate of recurrence and, more importantly, reducing the rate of progression . So far, bacille Calmette-Guerin is the only agent to decrease progression rate and thus increase survival . Bacille Calmette-Guerin has been shown to achieve this outcome by selectively sparing certain sequences during fibronectin degradation; these critical fragments were shown to have motility-stimulating activity, which may then enhance tumor invasion . In vivo and in vitro studies have shown that cathepsin B inhibitors prevent degradation of human basement membrane laminin, which is the first step of tumor invasion . Cathepsin B inhibitors either alone or in combination with bacille Calmette-Guerin may be valuable in preventing the progression of superficial bladder tumors in the future . Clinical studies on bacille Calmette-Guerin have repeatedly shown that a second course of bacille Calmette-Guerin to patients with local failure did not jeopardize patient survival . Adjuvant chemotherapy for patients with locally advanced disease and for those with regionally involved lymph nodes improves progression-free survival . Further controlled studies will establish the verdict for adjuvant chemotherapy . With the advancement of molecular genetic studies relevant to modern technology, our understanding of tumor behavior improves dramatically, and classic knowledge becomes more outdated every day. J Bacteriol, 1993 May, 175(10), 2952 - 60 Full expression of the cryIIIA toxin gene of Bacillus thuringiensis requires a distant upstream DNA sequence affecting transcription; de Souza MT et al.; The cryIIIA gene encoding a coleopteran-specific toxin is poorly expressed in Bacillus thuringiensis when cloned in a low-copy-number plasmid . This weak expression is observed when the gene is cloned only with its promoter and its putative terminator . cryIIIA gene expression was analyzed by using deletion derivatives of a larger DNA fragment carrying the toxin gene and additional adjacent sequences . The results indicate that a 1-kb DNA fragment located 400 bp upstream of the promoter strongly enhances CryIIIA production in B . thuringiensis sporulating cells . Similar results were obtained when the low-copy-number plasmid pHT304 carrying transcriptional fusions between upstream regions of cryIIIA and the lacZ gene was used . Analysis of the start sites, the sizes, and the amounts of cryIIIA-specific mRNAs shows that the enhancement occurs at the transcriptional level by increasing the number of cryIIIA-specific transcripts from the onset of sporulation to about 6 h after the onset of sporulation . The nucleotide sequence of the 1-kb activating fragment and of the 700 bp containing the promoter region and the 5' end of cryIIIA were determined . No potential protein-coding sequences were found upstream of the promoter . The major characteristic of the 1-kb activating fragment is the presence of a 220-bp A + T-rich region. J Infect Dis, 1993 May, 167(5), 1160 - 7 The major secreted antigen complex (Ag 85) from Mycobacterium bovis bacille Calmette-Guérin is associated with protective T cells in leprosy: a follow-up study of 45 household contacts; Launois P et al.; T cell proliferation and interferon (IFN)-gamma secretion were analyzed in 45 leprosy contacts stimulated with antigen 85 (Ag85), the major culture filtrate antigen from Mycobacterium bovis bacille Calmette-Guerin . All 14 Mitsuda reaction-positive contacts reacted to Mycobacterium leprae and Ag85 . Three Mitsuda reaction-negative contacts reacted weakly to M . leprae and Ag85 . The other 28 Mitsuda reaction-negative contacts did not react to M . leprae, but 9 reacted to Ag85 . Thirty-four contacts were retested 16 months later . Eleven contacts initially positive by the Mitsuda test remained lepromin positive and reactive to M . leprae and Ag85 . Fourteen contacts initially negative by the Mitsuda test converted, and all reacted in vitro to M . leprae and Ag85 . Finally, 9 contacts remained Mitsuda test-negative, and 7 were unreactive to Ag85 . In vitro reactivity to Ag85 at baseline in Mitsuda test-negative contacts was associated with subsequent conversion to lepromin reactivity in 7 of 9 subjects . These data suggest that reactive T cells against Ag85 develop very early during M . leprae infection and that Ag85 is a potentially protective T cell immunogen. J Appl Bacteriol, 1993 May, 74(5), 578 - 82 The application of laser diffractometry to study the water content of spores of Bacillus sphaericus with different heat resistances; De Pieri LA et al.; The distribution of water in the protoplast and integument of three populations of Bacillus sphaericus spores was determined by laser diffractometry together with the sizes of their integuments and protoplasts . The spores were produced at 15, 20 and 30 degrees C . Spores grown at 15 degrees C had protoplast and integument water contents similar to those produced at 20 degrees C, while those grown at 30 degrees C had significantly lower water contents than the other two populations . The inner (or protoplast) radii of the spores produced at 15, 20 and 30 degrees C were 0.41 +/- 0.02 microns, 0.42 +/- 0.01 microns and 0.38 +/- 0.02 microns whilst the outer (or whole spore) radii were 0.56 +/- 0.01 microns, 0.58 +/- 0.01 microns and 0.52 +/- 0.02 microns respectively . The ratios of integument to protoplast radius were 0.40 +/- 0.02, 0.43 +/- 0.07 and 0.41 +/- 0.03 respectively. Cancer Res, 1993 May 1, 53(9), 2123 - 7 Genetic construction, expression, and characterization of a single chain anti-carcinoma antibody fused to beta-lactamase; Goshorn SC et al.; We report the genetic construction and expression of a fusion protein between an antibody single chain-linked variable domain fragment specific for human carcinomas and beta-lactamase II from Bacillus cereus . Sequences encoding the variable regions of the L6 monoclonal antibody were assembled so as to be separated from each other by an 18-amino acid linker and from the mature form of beta-lactamase by a 6-amino acid linker . The construct was placed under the transcriptional regulation of the lac promoter, and the PelB signal sequence was used to direct export of the fusion protein to the periplasmic space of Escherichia coli . After induction, biologically active material was recovered from both culture supernatants and cell lysates . Affinity chromatography yielded about 2.5 micrograms of protein/ml of initial culture volume . The fusion protein was shown to bind to tumor cells at least as well as chemically prepared F(ab') and to maintain beta-lactamase activity at a level similar to that of the native enzyme . Tumor cells coated with the fusion protein were sensitive to a cephalosporin mustard prodrug in a dose-dependent fashion comparable to that of enzyme chemically conjugated to F(ab') . This article demonstrates the feasibility of using single chain-linked variable domain-enzyme fusion proteins for the activation of anticancer prodrugs. J Bacteriol, 1993 May, 175(9), 2762 - 6 Large-scale recrystallization of the S-layer of Bacillus coagulans E38-66 at the air/water interface and on lipid films; Pum D et al.; S-layer protein isolated from Bacillus coagulans E38-66 could be recrystallized into large-scale coherent monolayers at an air/water interface and on phospholipid films spread on a Langmuir-Blodgett trough . Because of the asymmetry in the physiochemical surface properties of the S-layer protein, the subunits were associated with their more hydrophobic outer face with the air/water interface and oriented with their negatively charged inner face to the zwitterionic head groups of the dipalmitoylphosphatidylcholine and dipalmitoylphosphatidylethanolamine (DPPE) monolayer films . The dynamic crystal growth at both types of interfaces was first initiated at several distant nucleation points . The individual monocrystalline areas grew isotropically in all directions until the front edge of neighboring crystals was met . The recrystallized S-layer protein and the S-layer-DPPE layer could be chemically cross-linked from the subphase with glutaraldehyde. Int J Biochem, 1993 May, 25(5), 689 - 96 Proof of alkaline phosphodiesterase I as a phosphatidylinositol-anchor enzyme; Nakabayashi T et al.; 1 . Ectoenzyme release from kidney brush border membranes of Rattus norvegicus and Sus scrofa domesticus by phosphatidylinositol-specific phospholipase C (PIPLC) of Bacillus thuringiensis was studied . 2 . The levels of specific activities of ectoenzymes in R . norvegicus kidney brush border membranes were higher than those in S . scrofa domesticus . About 10-fold higher values were found for specific activities of alkaline phosphatase and gamma-glutamyl transpeptidase in R . norvegicus . 3 . Alkaline phosphodiesterase I, alkaline phosphatase and 5'-nucleotidase were released from both R . norvegicus and S . scrofa domesticus brush border membranes, while gamma-glutamyl transpeptidase and dipeptidyl peptidase IV were not solubilized . The enzyme release by the action of PIPLC was suppressed when purified anti-PIPLC antibody was added to the reaction mixture . This suggests that enzyme release must be due to the direct action of PIPLC on kidney brush border membranes . 4 . The released alkaline phosphodiesterase I from kidney of S . scrofa domesticus had a molecular weight of 240,000 and was activated by Mg2+ and Ca2+, but strongly inhibited by EDTA. Res Immunol, 1993 May, 144(4), 233 - 43 Leishmania major infection in BALB/c mice: protection or exacerbation by treatment with different doses of BCG; Stefani MM et al.; The effect of live bacillus Calmette-Guerin (BCG), administered intraperitoneally to BALB/c mice, upon the development of lesions induced by subcutaneous infection with Leishmania major was examined . Lesions in mice given 10(7) BCG colony-forming units (CFU) 9 days before challenge with L . major were less severe and contained significantly fewer parasites than those of similarly infected control mice not given BCG . This effect of treatment with high doses of BCG upon the development of leishmanial lesions was observed using L . major promastigotes and amastigotes, whether or not 10(6) live BCG was included in the parasite inoculum . Lesions in mice given 5 x 10(4) BCG CFU 14 days before infection with L . major contained significantly fewer parasites than those of control mice not given BCG . Mice treated with low doses of BCG and infected with an L . major inoculum also comprising BCG exhibited larger lesions that contained more parasites . Interestingly, compared to naive mice infected with L . major, infection of naive mice with L . major mixed with live BCG consistently led to the development of more severe lesions that contained higher numbers of parasites . No correlation was found between the effect of BCG on the development of lesions induced by L . major and the amounts of IFN gamma, IL5 and TNF produced after in vitro antigenic challenge of either draining lymph node or spleen cells, the antigenic challenge being either live BCG or live L . major. J Invertebr Pathol, 1993 May, 61(3), 260 - 6 Use of synthetic peptides to probe functional domains of a Bacillus thuringiensis toxin; Pang AS; Two 10-residue peptides exhibiting sequence homology to CryIA(a) toxin were chemically synthesized . One corresponds to a segment from residues 48-57 (peptide A) and is common to all CryIA toxins and the second corresponds to a segment from residues 348-357 (peptide B) and is specific to the CryIA(a) toxin . Antibodies were raised in rabbits against both peptides . Antipeptide A did not protect either Bombyx mori or Choristoneura fumiferana larvae against CryIA(a) toxin . However, antipeptide B offered significant protection to B . mori and somewhat less protection to C . fumiferana larvae . Neither antiserum interfered with the binding of CryIA(a) toxin to brush border membrane vesicles of either insect . The results suggest that the segment from residues 348-357 in CryIA(a) toxin is important for expression of toxicity in both insects but not for binding. Mikrobiol Zh, 1993 May-Jun, 55(3), 104 - 10 {The role of Bacillus thuringiensis in natural biocenoses}; Golovko AE et al.; The results of the author's data and those from literature on ecology and genetic exchange of Bacillus thuringiensis strains have been analyzed in this brief review . It has been concluded that there is no strict confinement of B . thuringiensis strains of the particular serotypes to certain habitat and that the bacteriocinogenicity is of great importance in the intraspecies competition and stability of strains in nature . It has been emphasized that the plasmid conjugation process in B . thuringiensis can serve as a main factor of genetic variability to establish ecological placement of these bacteria. Rinsho Byori, 1993 May, 41(5), 543 - 7 {The role of the clinical microbiology laboratory in the past}; Kudo H; Over the past 150 years, since Koch first isolated the tubercle bacillus, clinical microbiologists have used culturing techniques to demonstrate of the presence of pathogenic microorganisms associated with infectious diseases in clinical specimens . The development of medicine and chemotherapy and the improvement of sanitary conditions resulted in a marked decrease in infectious diseases . In stead of infections due to virulent pathogens, opportunistic infections in the compromised hosts by non-virulent or weakly virulent bacteria have been on the rise . As a result, determining the pathogenicity of an isolated bacteria becomes important . The past two decades have seen extensive efforts to exploit the potential of automation in clinical microbiology and to develop increasingly rapid procedures . New technology, such as molecular genetics, has also been introduced into clinical microbiology . For more correct and precise diagnoses of infectious diseases, the clinical microbiology laboratory should be enlarged as an "Infectious Disease Laboratory" to include an extended work on not only the detection of pathogens from the specimens of infectious disease, but also serodiagnostic tests and immunological tests of hosts . The "Infectious Disease Laboratory" would also play a key role in the control of nosocomial infection working with the infection control committee and infectious disease clinicians. J Biochem (Tokyo), 1993 May, 113(5), 607 - 13 Purification and cDNA cloning of bovine liver 5'-nucleotidase, a GPI-anchored protein, and its expression in COS cells; Suzuki K et al.; A glycosylphosphatidylinositol (GPI)-anchored protein, 5'-nucleotidase {EC 3.1.3.5}, was released from the membrane of bovine liver by use of phosphatidylinositol-specific phospholipase C (PI-PLC) of Bacillus thuringiensis and purified by several column chromatographies to a homogeneous state . The purified protein has an apparent molecular mass of 61 kDa, as estimated by SDS-polyacrylamide gel electrophoresis . From the partial amino acid sequence of a tryptic peptide, mixed oligonucleotides were synthesized and used to screen a lambda gt11 liver cDNA library, and one positive clone, pE1, was isolated . Since the insert of the clone lacked the NH2-terminal coding region, another lambda gt11 liver cDNA library was screened by using a synthetic probe corresponding to the 5' region of the insert of pE1 . Three additional cDNA clones were obtained . Sequencing of these cDNAs revealed an open reading frame that encodes a 574-residue polypeptide with a calculated mass of 63,084 Da . The predicted structure showed two highly hydrophobic stretches at both ends of the protein, like those of rat and human 5'-nucleotidases . The NH2-terminal 26 residues comprise a signal peptide and the COOH-terminal hydrophobic stretch may serve as a signal for the posttranslational GPI modification . An expression vector of the cDNA, pSVNT, was constructed in a mammalian expression vector pSVL and the 5'-nucleotidase activity was transiently expressed in COS-1 cells . The expressed activity was about 8 times higher than the pSVL-transfected control activity . PI-PLC released 45% of the transiently expressed 5'-nucleotidase activity, indicating that the cDNA isolated here encodes this enzyme expressed as a GPI-anchored protein. Hinyokika Kiyo, 1993 May, 39(5), 463 - 5 {Kidney-conservative surgery followed by bacillus Calmette-Guerin instillation therapy for transitional cell carcinoma of solitary renal pelvis: a case report}; Ao T et al.; We report a case (62-year-old male) of transitional cell carcinoma (grade 2) of solitary right renal pelvis which was successfully treated with tumor excision followed by bacillus Calmette-Guerin instillation therapy . He had a past history of left radical nephroureterectomy for renal pelvic tumor 11 years earlier . Kidney-preserved surgery with BCG instillation therapy is expected to be an alternative for radical forms of therapy, especially in the patients with a solitary kidney. Anticancer Res, 1993 May-Jun, 13(3), 705 - 8 The anti-metastatic effect of intravenously-inoculated BCG on prostate tumor cells; Pollard M et al.; The complication of metastasis from the primary tumor site to the distant organ is difficult to control . Tumor cells usually spread through the vascular system: lymphatics and/or blood; or by direct extension into adjacent sites . Transplantable prostate adenocarcinoma (PA-III) cells in L-W rats produces a tumor in the implant site and then spreads via the ipsilateral lymphatic route to the lungs in which new visible tumors develop . Viable bacillus Calmette-Guerin (BCG) was inoculated SC or IV into rats which were then inoculated either SC or IV with PA-III cells . They were examined thereafter for primary and for lung tumors . IV-inoculated BCG generated a strong intravascular intervention mechanism on metastatic PA-III cells in L-W rats . This immobilization mechanism was not demonstrable in rats that had been inoculated SC with BCG. Res Microbiol, 1993 May, 144(4), 271 - 8 Immunological localization of Bacillus thuringiensis serovar israelensis toxins in midgut cells of intoxicated Anopheles gambiae larvae (Diptera: Culicidae); Ravoahangimalala O et al.; Fourth instar larvae of Anopheles gambiae were intoxicated with doses of purified crystals from Bacillus thuringiensis serovar israelensis (Bti) corresponding to 50-fold the LC50 after 24 h . Midguts were dissected after various contact times, then processed for immuno-light and -electron microscopy . Immunodetection on thin sections was performed using affinity-purified rabbit IgG against Bti crystal CryIVD or CytA polypeptides, in combination with anti-rabbit IgG/peroxidase . Both polypeptides were detected by optical and electron microscopy after 15 min of contact with Bti crystals on the apical brush border of midgut cells, but only in the gastric caeca and posterior stomach . No specific signal was detected in the other parts of the midgut, i.e . the cardia cells and the anterior stomach . These results confirm that mosquito midgut cells are the primary target for the toxins and that binding to specific receptors on the apical microvilli membrane is the initial step of delta-endotoxin action. Nippon Saikingaku Zasshi, 1993 May, 48(3), 541 - 50 {Fragmentation and solubilization of filamentous appendages of Bacillus cereus spores}; Kozuka S; Filamentous appendages were removed from the surfaces of Bacillus cereus spores by treatment with sodium thioglycolate and purified by filtration through a glass microfiber paper and a membrane filter . The basic structure of the isolated appendages resembled that of the native appendages on the spores . The results of electron microscopical analysis, amino acid analysis, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) attested the purity of the preparation . The purified filamentous appendages were fragmented by treatment with 2N sodium hydroxide at 30C but neither with SDS nor mercaptoethanol, and the fragmentation progressed with the prolongation of the treatment period . When observed under an electron microscope, each fragmented appendage appeared to be rod shaped with an apparent axial hole and helical or disk-like arrangement of subunits . In the agar gel immunodiffusion test, a soluble fraction of the appendages obtained by treatment with sodium hydroxide formed only a single precipitation line against the antibody to the intact appendages . The molecular mass of the subunit separated by SDS-PAGE was approximately 10kDa . The 10-kDa protein was distributed over the surface of the intact appendages in immunoelectron microscopy . The antigenicity of the appendage subunits disappeared upon treatment with 2N sodium hydroxide for longer than 24h . Neither intact appendages nor intact spores agglutinated sheep or guinea-pig erythrocytes. Appl Microbiol Biotechnol, 1993 May, 39(2), 197 - 203 Cloning and sequence analysis of the cyclomaltodextrinase gene from Bacillus sphaericus and expression in Escherichia coli cells; Oguma T et al.; The gene for cyclomaltodextrinase (CDase; EC 3.2.1.54) from Bacillus sphaericus E-244 was cloned in the recombinant plasmid pCD629 . Sequencing a portion of pCD629 revealed a unique open reading frame of 1,773 nucleotides coding for a 591-amino-acid polypeptide . The deduced polypeptide sequence showed about 50% homology with that of a neopullulanase, and was slightly homologous to those of the cyclodextrin glucanotransferases and the alpha-amylases . The optimum pH, specific activity and Km value for beta-cyclodextrin of the CDase that has been produced in Escherichia coli cells were 8.0, 16.4 units/mg protein, and 0.41 mM, respectively . These values were almost identical to those from B . sphaericus E-244. J Biochem (Tokyo), 1993 May, 113(5), 637 - 41 Induction of antibiotic production by protease in Bacillus brevis (ATCC 8185); Oyama M et al.; Bacillus brevis (ATCC 8185) produces an antibiotic peptide, linear gramicidin, in the early stationary growth phase . Since we observed that preculture in milk medium is essential for production of the antibiotic in broth medium, we studied the role of the preculture in the antibiotic production . We found that addition of the supernatant of the precultured milk medium was sufficient to induce gramicidin production in broth medium . Fresh milk medium had no effect . The effector substance in the overnight cultured milk medium was labile at both acidic and alkaline pH and was destroyed by heat . We also found that addition of a protease (for example, bovine pancreas chymotrypsin or Streptomyces griseus protease) instead of the supernatant could induce the gramicidin production . Addition of Mn2+ was not required for the protease-induced production of gramicidin . It is known that B . brevis cells secrete protease into milk medium . But neither use of the protease-pretreated broth medium nor addition of casamino acids to broth medium induced gramicidin production . These results suggest that B . brevis cells secrete a factor for linear gramicidin production, that the inducing factor is protease and that the target of the protease is a substance(s) produced by the bacteria. J Am Vet Med Assoc, 1993 Apr 15, 202(8), 1249 - 54 Field trial of four cowside antibiotic-residue screening tests; Sischo WM et al.; Four commercially available screening tests for antibiotic residues in milk were evaluated for their ability to correctly identify the antibiotic status of cows . A field trial, which included 199 cows from 2 herds, was conducted . Sensitivity, specificity, and likelihood ratios for a positive test result were calculated by using the Bacillus stearothermophilus var calidolactis disk assay as the reference test . The relationship of risk factors to the probability of a false-positive result for each screening test was modeled by use of unconditional logistic regression . The risk factors evaluated in these models were loge somatic cell count (scc), intramammary infection, herd, milk appearance, time milk sample frozen before tested, days in lactation, parity, and manufacturer's lot number . The risk factors log(e) scc and intramammary infection were forced into all models . The overall specificities for the 4 tests ranged from 0.78 to 0.95, whereas likelihood ratios for a positive test result ranged from 4.54 to 20.0 . When the confounding of cofactors was controlled in the logistic model, there was a positive effect of log(e) scc on the probability of a false-positive result for 3 of the screening tests, that is, for incremental increases in log(e) scc, there was an increasing likelihood for a false-positive result . In some of the tests, parity and intramammary infection also influenced the likelihood of a false-positive result . The goal of cowside testing is to assist in the production of high-quality, antibiotic residue-free milk from dairies.(ABSTRACT TRUNCATED AT 250 WORDS) J Biol Chem, 1993 Apr 15, 268(11), 8240 - 5 Evidence that the CryIA crystal protein from Bacillus thuringiensis is associated with DNA; Bietlot HP et al.; Toxin generated by activation of the Bacillus thuringiensis CryIA(c) crystal protein (protoxin) with bovine trypsin was separated into two components by anion-exchange chromatography . One component (T2) was DNA-associated toxin, and the other was the DNA-free toxin (T1) . Only one major protoxin component was observed, and it was found to be associated with DNA . The DNA from the T2 toxin varied in size from 100 to 300 base pairs, whereas the crystal and the solubilized protoxin contained 20-kilobase DNA as the major DNA component . DNase treatment converted the T2 toxin to the DNA-free T1 toxin . In contrast, the DNA in the crystal and the solubilized protoxin was resistant to DNase digestion and was not dissociated from the protein by 1.5 M NaCl . The protoxin and DNA appeared to elute as a complex with a molecular mass of > 2 x 10(6) Da on gel-filtration chromatography . No toxin was generated from the protoxin with trypsin after extensive digestion of the protoxin with DNase or dissociation of the DNA by succinylation of the lysine residues . It is proposed that DNA binds to the COOH-terminal half of the crystal protein and is essential for maintaining the conformational integrity required for crystal formation and generation of toxin. Eur J Biochem, 1993 Apr 15, 213(2), 683 - 7 Identification of charge-transfer transitions in the optical spectrum of low-spin ferric cytochrome P-450 Bacillus megaterium; McKnight J et al.; The optical, low temperature magnetic circular dichroism (MCD) and EPR spectra of low-spin Fe(III) cytochrome P-450 BM-3 from Bacillus megaterium, and its imidazole adduct have been measured . The MCD spectra locate new transitions over 600-700 nm and 800-1300 nm . The latter are assigned to porphyrin (a1u)-Fe(III) (dyz) charge-transfer (CT) transitions . In the case of the imidazole adduct the energy of this transition fits well to the theoretical model of Gadsby and Thomson {Gadsby, P . M . A . & Thomson, A . J . (1990) J . Amer . Chem . Soc 112, 5003-5011} . For the native enzyme, the energy of the CT band suggests co-ordination by a strongly H-bonded water ligand and the axial thiolate form of cysteine . Two transitions between 600-700 nm are detected in both derivatives . A theoretical analysis and fit of the MCD magnetisation properties shows that these transitions are polarised XY and XZ, respectively . They are assigned as thiolate sulphur py-d-shell and pz-d-shell CT transitions, analogous to the well known 695 nm band of methionine-histidine co-ordinated haem as in cytochrome c . They should prove usefully diagnostic of cysteine/Fe(III) conformational changes or protonation. Carbohydr Res, 1993 Apr 7, 242, 173 - 80 Identification of 2-amino-2,6-dideoxy-D-glucose (D-quinovosamine), isolated from the cell walls of the alkaliphilic Bacillus sp . Y-25, by 500-MHz 1H NMR spectroscopy; Ito M et al.; Cell walls of alkaliphilic Bacillus strain Y-25 are composed of gamma-peptidoglycan and two acidic polymers . An amino sugar, which was a min component of one acidic polymer, did not correspond to any of the commercially available hexosamines . The amino sugar was isolated from the hydrolysate of the acidic polymer, purified, and identified as D-quinovosamine (2-amino-2,6- dideoxy-D-glucose) by 500-MHz NMR spectroscopic analysis and polarimetry. Biochemistry, 1993 Apr 6, 32(13), 3429 - 36 Structural and functional characterization of the alpha 5 segment of Bacillus thuringiensis delta-endotoxin; Gazit E et al.; One of the most conserved sequences in various delta-endotoxins is the 30 amino acid long block I . Block I of cryIIIA delta-endotoxin contains a 23 amino acid amphiphilic alpha-helix termed alpha 5 . The potential involvement of this alpha 5 helix in the toxic mechanism of delta-endotoxin was examined . For this purpose, a peptide corresponding to the alpha 5 segment and its proline incorporated analogue (P-alpha 5) were synthesized and characterized . The alpha-helical content of the peptides, assessed in methanol by circular dichroism (CD), was 58% and 24% for alpha 5 and P-alpha 5, respectively . To monitor the interaction of alpha 5 peptides with phospholipid membranes, they were selectively labeled at their N-terminal amino acids with the fluorescent probes 7-nitrobenz-2-oxa-1,3-diazol-4-yl (NBD) or carboxyfluorescein . Fluorometric studies allowed the calculation of membrane surface partition constants, which were about 10(4) M-1 for both alpha 5 and P-alpha 5, and revealed that their N-terminals are located within the lipid bilayers . The shape of the binding isotherms indicated that alpha 5 aggregated in both zwitterionic and acidic vesicles . Functional characterization of the alpha 5 peptides was determined by assessing their ability to dissipate a diffusion potential from sonicated small unilamellar vesicles (SUV) composed of zwitterionic or acidic phospholipids and to lyse human erythrocytes . alpha 5 was much more active than P-alpha 5 in both assays . Moreover, membrane-bound alpha 5 was more protected from enzymatic proteolysis than P-alpha 5.(ABSTRACT TRUNCATED AT 250 WORDS) J Biol Chem, 1993 Apr 5, 268(10), 7553 - 61 Critical residues involved in FMN binding and catalytic activity in cytochrome P450BM-3; Klein ML et al.; Cytochrome P450BM-3 from Bacillus megaterium is a soluble, catalytically self-sufficient fatty acid mono-oxygenase that, in structural organization and amino acid sequence, resembles the Class II (microsomal) P450 systems . Its single polypeptide chain contains both a P450 heme domain and an NADPH:P450 reductase domain, each of which bears significant homology with its microsomal counterparts . We report here the critical nature of three amino acids in the reductase domain of this enzyme with respect to FMN binding and catalytic activity . We used site-directed mutagenesis to change glycine 570 to bulkier amino acids; none of these mutant enzymes contained FMN after purification . We also made substitutions for tryptophan 574 and tyrosine 536, which by sequence analogy (Porter, T . D . (1991) Trends Biochem . Sci . 16, 154-158) were proposed to bind FMN through stacking of the aromatic rings with the isoalloxazine ring of the flavin . Mutants of tryptophan 574 which retained the aromatic side chain contained no less than 0.85 mol of FMN per mol of enzyme, while aspartate and glycine substitutions yielded enzymes which did not incorporate FMN . Substitution of tyrosine 536 with aspartate gave an enzyme which contained 0.44 mol of FMN per mol of enzyme but was inactive as a fatty acid hydroxylase and had only 2% of wild-type cytochrome c reductase activity, while the glycine mutant at this position bound no FMN . Furthermore, although all of the mutant enzymes contained 1 mol of FAD per mol of enzyme, the Y536D mutant and those entirely lacking FMN retained no more than 40% of wild-type ferricyanide reductase activity . By assaying these enzymes in the presence of added FMN, we were able to assess the relative importance of the residues in the wild-type sequence with respect to their contribution to FMN binding . In addition, the aromatic mutants of tryptophan 574, which were nearly as active in cytochrome c reduction as wild-type P450BM-3, were only 20% as active in myristate hydroxylation as the wild-type enzyme, suggesting that this amino acid plays an important role in the flow of electrons between the P450 heme and reductase domains. J Biol Chem, 1993 Apr 5, 268(10), 6932 - 8 Kinetic and stereochemical comparison of wild-type and active-site K145Q mutant enzyme of bacterial D-amino acid transaminase; Bhatia MB et al.; D-Amino acid transaminase (EC 2.6.1.21), from Bacillus sp . YM-1, a thermostable enzyme with pyridoxal 5'-phosphate as coenzyme and a target for the design of novel antimicrobial agents, catalyzes the reversible transfer of an amino group between D-alanine and alpha-ketoglutarate to form pyruvate and D-glutamate, respectively . To explore the catalytic role of Lys-145, which binds the coenzyme, a site-specific mutant enzyme, K145Q (in which Lys-145 had been mutated to glutamine) constructed earlier (Futaki, S., Ueno, H., Martinez del Pozo, A., Pospischil, M . A., Manning, J . M., Ringe, D., Stoddard, B., Tanizawa, K., Yoshimura, T., and Soda, K . (1990) J . Biol . Chem . 265, 22306-22312) was compared to the wild-type enzyme for its kinetic parameters . Initial velocity studies and partial reaction isotope exchange experiments showed that the low activity of the mutant enzyme (about 1.5% the activity of the wild-type enzyme with saturating substrates) is an intrinsic property, confirming that contaminating enzymes do not account for the low activity of the K145Q mutant enzyme . The rates of the forward reaction for both wild-type and mutant enzymes were 30-40 times higher than the rates of the reverse reaction . KM values for the four substrates were 10 to 100 higher for the mutant compared to the wild-type enzyme . Whereas D-alanine is preferred over L-alanine by the wild-type enzyme (10(3) higher kcat/KM for D- over L-alanine), the K145Q enzyme does not efficiently discriminate between L- and D-alanine . Both wild-type and mutant enzymes also catalyze the slow racemization of L- and D-alanine . Proton NMR studies showed that wild-type enzyme catalyzed a time-dependent exchange of the C alpha proton of D-alanine with solvent D2O and a slow exchange of the alpha proton of L-alanine; the latter slow exchange rate is the same for the C alpha proton of both L- and D-alanine with the K145Q mutant enzyme . Thus, in addition to binding pyridoxal 5'-phosphate, the active-site Lys-145 of D-amino acid transaminase is involved in several other important functions, i.e . it optimizes catalytic efficiency and it maintains stereochemical fidelity . The steady-state kinetic results on the K145Q mutant enzyme together with the findings on the relative racemization rates and the NMR protein exchange data suggest that an alternate base catalyzes abstraction of the alpha proton of substrate in this mutant D-amino acid transaminase. J Gen Microbiol, 1993 Apr, 139 ( Pt 4), 775 - 80 Properties of the cold-labile NAD(+)-specific glutamate dehydrogenase from Bacillus cereus DSM 31; Jahns T et al.; Nicotinamide-adenine-dinucleotide-specific glutamate dehydrogenase (NAD-GDH; EC 1.4.1.3) from Bacillus cereus DSM 31 was enriched 260-fold . The molecular mass was determined by gel filtration to be 270 kDa (+/- 25 kDa) . The enzyme was highly specific for the coenzyme NAD(H) and catalysed both the formation and the oxidation of glutamate . Apparent Km values of 7.7 mM for glutamate and 0.56 mM for NAD+ during oxidative deamination were measured . Both in crude cell-free extracts and in enriched preparations the enzyme was extremely unstable, especially at low temperatures . The loss of activity in the cold was found to be due to the dissociation of the holoenzyme into catalytically inactive subunits of molecular mass 48 kDa (+/- 5 kDa), indicating that the native enzyme has a hexameric structure . The activity was restored under certain conditions, and no instability of the enzyme in the cold was observed in undisrupted cells. An Med Interna, 1993 Apr, 10(4), 188 - 94 {Prevention of tuberculosis infection}; Prados C et al.; Tuberculosis (TB) is a disease caused by a mycobacterium, whose incidence has increased in the past years . This increase is related to the adquired immunodeficiency syndrome (AIDS) . Due to its high prevalence, Spain is considered a developing country . The tuberculous infection depends on the degree of functionality of the alveolar macrophages that stimulate the lymphocytes and isolate the bacillus . The infection by mycobacterias can be quantified by means of the cutaneous reaction against tuberculin and mantoux, allowing us to select the subjects that must receive prophylaxis . For its correct interpretation, it is currently recommended to avoid BCG vaccination of children, except in countries with high prevalence of TB. J R Coll Physicians Lond, 1993 Apr, 27(2), 169 - 74 Microbes, molecules and man . The Mitchell Lecture 1992; Moxon ER; Robert Koch, the discoverer of the tubercle bacillus, has had a seminal influence on the extraordinary progress in the field of infectious diseases in the past 100 years . Koch's postulates defined the germ theory of disease causation . They have now been confirmed and brought up to date by the application of molecular techniques . Developments in molecular genetics have helped in the elucidation of the pathogenesis of meningitis caused by H influenzae and the role of type b capsule as a determinant of bacterial virulence--and satisfied the requirements of Koch's molecular postulates . This new knowledge has contributed to the development of a successful immunoprophylactic strategy for eliminating Hib disease . Studies in Oxford over the past eight years have confirmed the effectiveness and safety of a routine immunisation programme for the UK. Int J Syst Bacteriol, 1993 Apr, 43(2), 221 - 31 Characterization of Bacillus brevis with descriptions of Bacillus migulanus sp . nov., Bacillus choshinensis sp . nov., Bacillus parabrevis sp . nov., and Bacillus galactophilus sp . nov; Takagi H et al.; Thirty-five Bacillus brevis strains obtained from culture collections, including protein-producing isolates, were taxonomically studied by using numerical analysis, DNA base composition, and DNA-DNA hybridization . Six DNA relatedness groups were represented, and these groups correlated well with clusters based on the numerical analysis . The B . brevis strains were separated into B . brevis sensu stricto, four new species, and an unidentified species of the genus Bacillus . Bacillus migulanus sp . nov., Bacillus choshinensis sp . nov., Bacillus parabrevis sp . nov., and Bacillus galactophilus sp . nov . are proposed. Nippon Hinyokika Gakkai Zasshi, 1993 Apr, 84(4), 656 - 61 {Long-term results and risk factors of tumor recurrence in patients with superficial bladder cancer who were treated by intravesical bacillus Calmette-Guerin (BCG) instillation}; Tachibana M et al.; An evaluation was made of the long-term results and risk factors of tumor recurrence in patients with superficial bladder cancer who were treated with intravesical bacillus Calmette-Guerin (BCG) and who had a mean follow-up period of 57 months . Eligible for the study were a total of 102 patients who were treated by transurethral resection of tumors and were considered to be free from tumor from 1984 to 1989 . A suspension containing 80 mg Tokyo 172 strain BCG in 50 ml normal saline was given intravesically to 50 patients once a week for six weeks without further maintenance instillation . To the other 52 patients was additionally given monthly intravesical BCG instillation for 12 months . The actuarial nonrecurrent rates according to sex, different BCG treatment protocol, tumor grade, tumor status including primary or recurrent tumors, and solitary or multiple tumors were estimated by Kaplan-Meier method . The estimated three-, five- and seven-year actuarial nonrecurrent rates in all 102 cases were 77.3%, 68.5% and 60.6%, respectively . When nonrecurrent rates were compared to tumor characteristics, no statistically significant differences were observed between primary and recurrent tumors or between solitary and multiple tumors estimated by generalized Wilcoxon method . On the other hand, when nonrecurrent rates were compared with tumor grades, grade 3 tumor showed a 40.0% three-year nonrecurrent rate . The differences between grade 3 and grade 1 and between grade 3 and grade 2 were statistically significant (p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) FEMS Microbiol Lett, 1993 Apr 1, 108(2), 139 - 43 Expression of bacteriophage PhiX174 lysis gene E in Staphylococcus carnosus TM300; Halfmann G et al.; Expression of the cloned PhiX174 gene E causes lysis of the Gram-negative bacterium Escherichia coli, which led to the proposal that a two-membrane system is necessary for the protein E lysis function . Gene E was cloned in an E . coli/Bacillus subtilis shuttle vector and expressed in the Gram-positive bacterium Staphylococcus carnosus TM300 . Regulated gene E expression had a lethal effect on S . carnosus; however, no lysis was detected, lending support to the hypothesis. Biotechnol Appl Biochem, 1993 Apr, 17 ( Pt 2), 205 - 16 Cyclomaltodextrin glucanotransferase from Bacillus circulans E 192: nitration with tetranitromethane; Villette JR et al.; Nitration of tyrosine residues was performed on Bacillus circulans E 192 cyclomaltodextrin glucanotransferase (CGTase) using tetranitromethane (TNM) . A maximum of 15 out of 28 tyrosine residues is modified with 8 mM TNM, entailing a concomitant loss of enzymic activity and tryptophan fluorescence . Spectroscopic studies suggest that these two phenomena are related to an impairment of the enzyme conformation as a consequence of the tyrosine nitration . The presence of 5 mM acarbose during the CGTase nitration results in the protection of one tyrosine residue and the rate of inactivation is reduced 9.4-fold . These results support a contribution of a tyrosine residue in the CGTase catalytic site . The nitration of CGTase also entails a decrease in the enzyme's affinity for a beta-cyclodextrin (beta-CD) co-polymer . Kinetic and analytical investigations on isolated modified enzymes support the concept that this phenomenon is unrelated to the modification of tyrosine residues, but rather concerns a side reaction of the reagent occurring at the raw-starch-binding site of the CGTase. Appl Environ Microbiol, 1993 Apr, 59(4), 1253 - 8 Expression and secretion of Bacillus amyloliquefaciens alpha-amylase by using the yeast pheromone alpha-factor promoter and leader sequence in Saccharomyces cerevisiae; Southgate VJ et al.; Replacement of the regulatory and secretory signals of the alpha-amylase gene (AMY) from Bacillus amylolique-faciens with the complete yeast pheromone alpha-factor prepro region (MF alpha 1p) resulted in increased levels of extracellular alpha-amylase production in Saccharomyces cerevisiae . However, the removal of the (Glu-Ala)2 peptide from the MF alpha 1 spacer region (Lys-Arg-Glu-Ala-Glu-Ala) yielded decreased levels of extracellular alpha-amylase. Appl Environ Microbiol, 1993 Apr, 59(4), 1131 - 7 Cloning of a novel cryIC-type gene from a strain of Bacillus thuringiensis subsp . galleriae; Kalman S et al.; A novel cryIC-type gene was isolated from a strain of Bacillus thuringiensis subsp . galleriae . A new polymerase chain reaction (PCR) technique with a set of several oligonucleotide primer pairs specific to the cryIC gene was used to screen a number of B . thuringiensis strains . PCR amplified several DNA fragments ranging from 100 bp to 1 kb for B . thuringiensis strains containing a cryIC gene . PCR fragments amplified from the Bacillus thuringiensis subsp . galleriae HD29 DNA differed from the fragments amplified from other cryIC-containing strains, indicating strain HD29 contained a novel cryIC-type gene . To isolate crystal genes homologous to cryIC, an HD29 gene library was probed with a 984-bp fragment of the amino-terminal coding region of the cryIC gene cloned from Bacillus thuringiensis subsp . aizawai HD229 . A putative toxin gene was isolated from a phage that hybridized strongly to the cryIC probe . Translation of the putative toxin DNA sequence revealed an open reading frame of 1,176 amino acids whose predicted molecular mass was 132.8 kDa . Comparisons of the toxin gene sequence with sequences of other cry genes indicated that this gene is a subclass of cryIC . We propose to designate this gene cryIC(b) . In Escherichia coli, the cryIC(b) gene produced a protein of approximately 130 kDa toxic to Spodoptera exigua and Trichoplusia ni. Biochem J, 1993 Apr 1, 291 ( Pt 1), 151 - 5 An overview of the kinetic parameters of class B beta-lactamases; Felici A et al.; The catalytic properties of three class B beta-lactamases (from Pseudomonas maltophilia, Aeromonas hydrophila and Bacillus cereus) were studied and compared with those of the Bacteroides fragilis enzyme . The A . hydrophila beta-lactamase exhibited a unique specificity profile and could be considered as a rather specific 'carbapenemase' . No relationships were found between sequence similarities and catalytic properties . The problem of the repartition of class B beta-lactamases into sub-classes is discussed . Improved purification methods were devised for the P . maltophilia and A . hydrophila beta-lactamases including, for the latter enzyme, a very efficient affinity chromatography step on a Zn(2+)-chelate column. Vet Hum Toxicol, 1993 Apr, 35(2), 141 - 3 Sump additives as a source of bioaerosols in a school building; Thorne PS; An investigation was launched following complaints of poor air quality and building-related illness in a public elementary school . Occlusion of air intakes put the building under negative pressure and caused vents from a below-ground sump to become air intakes . Outside air drawn through the sump pit traveled into the adjacent main air handling unit and was disseminated throughout the building . Sump additives introduced in an attempt to counteract foul odors contained spores of Bacillus species, which appeared as bioaerosols throughout the school . Viable microbial sampling identified B subtilis, B cereus, and B licheniformis in the sump room and classrooms at levels as high as 760 colony forming units/m3 (CFU/m3) . Concentrations of CO2 in classrooms were 1250 ppm, indicating inadequate makeup air . Remediation was accomplished by opening the air intakes, isolating the sump room from the air handling system, venting the sump to the outside, and flushing the sump with fresh water on a regular basis. J Bacteriol, 1993 Apr, 175(8), 2271 - 7 Cloning and nucleotide sequence of the Myxococcus xanthus lon gene: indispensability of lon for vegetative growth; Tojo N et al.; The lon gene of Escherichia coli is known to encode protease La, an ATP-dependent protease associated with cellular protein degradation . A lon gene homolog from Myxococcus xanthus, a soil bacterium which differentiates to form fruiting bodies upon nutrient starvation, was cloned and characterized by use of the lon gene of E . coli as a probe . The nucleotide sequence of the M . xanthus lon gene was determined . It contains an open reading frame that encodes a 92-kDa protein consisting of 817 amino acid residues . The deduced amino acid sequence of the M . xanthus lon gene product showed 60 and 56% identity with those of the E . coli and Bacillus brevis lon gene products, respectively . Analysis of an M . xanthus strain carrying a lon-lacZ operon fusion suggested that the lon gene is similarly expressed during vegetative growth and development in M . xanthus . In contrast to that of E . coli, the M . xanthus lon gene was shown to be essential for cell growth, since a null mutant could not be isolated. J Bacteriol, 1993 Apr, 175(8), 2248 - 54 Relevance of charged groups for the integrity of the S-layer from Bacillus coagulans E38-66 and for molecular interactions; Sara M et al.; In this paper, the importance of charged amino and carboxyl groups for the integrity of the cell surface layer (S-layer) lattice from Bacillus coagulans E38-66 and for the self-assembly of the isolated subunits was investigated . Amidination of the free amino groups which preserved their positive net charge had no influence on both . On the other hand, acetylation and succinylation, which converted the amino groups into either neutral or negatively charged groups, and amidation of carboxyl groups were accompanied by the disintegration or at least by the loss of the regular structure of the S-layer lattice . Treatment of S-layer monolayers with the zero-length cross-linker carbodiimide led to the introduction of peptide bonds between activated carboxyl groups and amino groups from adjacent subunits . This clearly indicated that in the native S-layer lattice the charged groups are located closely enough for direct electrostatic interactions . Under disrupting conditions in which the S-layer polypeptide chains were unfolded, 58% of the Asx and Glx residues could be amidated, indicating that they occur in the free carboxylic acid form . As derived from chemical modification of monolayer self-assembly products, about 90% of the lysine and 70% of the aspartic and glutamic acid residues are aligned on the surface of the S-layer protein domains . This corresponded to 45 amino groups and to 63 carboxyl groups per S-layer subunit . Labelling experiments with macromolecules with different sizes and charges and adsorption studies with ion-exchange particles revealed a surplus of free carboxyl groups on the inner and on the outer faces of the S-layer lattice . Since the carboxyl groups on the outer S-layer face were accessible only for protein molecules significantly smaller then the S-layer protomers or for positively charged, thin polymer chains extending from the surface of ion-exchange beads, the negatively charged sites must be located within indentations of the corrugated S-layer protein network . This was in contrast to the carboxyl groups on the inner S-layer face, which were found to be exposed on elevations of the S-layer protein domains (D . Pum, M . Sara, and U.B . Sleytr, J . Bacteriol . 171:5296-5303, 1989). Am Rev Respir Dis, 1993 Apr, 147(4), 958 - 61 Quantitative bacillary response to treatment in HIV-associated pulmonary tuberculosis; Brindle RJ et al.; A group of 122 patients with culture-proven pulmonary tuberculosis were recruited to examine the concentrations of Mycobacterium tuberculosis in sputum and the relationship to HIV-1 antibody status . They were followed for up to 28 days from the start of antituberculous chemotherapy to assess the early bacillary response to two chemotherapeutic regimens . Of 67 treated with streptomycin, thiacetazone, and isoniazid 17 were HIV positive, and subsequently 55, of whom 20 were HIV positive, were treated with streptomycin, rifampin, isoniazid, and pyrazinamide . The mean initial concentration of M . tuberculosis in the sputum of the HIV-negative patients was significantly higher than in HIV-positive patients (6.95 and 6.34 log colony-forming units respectively; p = 0.019) . The HIV-positive patients had less radiologic evidence of disease and significantly fewer zones of lung affected with cavities . The response to treatment was similar, but with HIV-positive patients more likely to become culture negative by 28 days . The differences that exist between HIV-positive and HIV-negative patients are minor, and standard regimens are at least as effective in HIV-positive patients in the first month of treatment. J Clin Microbiol, 1993 Apr, 31(4), 972 - 4 Rapid screening test for enterotoxin-producing Bacillus cereus; Jackson SG; Culture supernatants of 30 enterotoxin-producing Bacillus cereus isolates produced a characteristic progressive destruction of McCoy cell monolayers . Enterotoxin-negative B . cereus and other group 1 Bacillus spp . caused no monolayer disruption . The McCoy cell tissue culture system appears to provide a rapid screening assay for detection of enterotoxin-producing B . cereus. Am J Public Health, 1993 Apr, 83(4), 583 - 5 Complications of BCG vaccinations in rural Haiti; Bonnlander H et al.; This study investigated an outbreak of axillary lymphadenitis and abscesses after Bacillus Calmette-Guerin vaccination among rural Haitian children treated at the Hospital Albert Schweitzer from January 1986 through March 1991 . Seventy-seven cases of vaccine-related complications were identified, all among children immunized before the age of 1 year . The proportions of children with complications were 0.017% for 1986 through 1989, 0.91% for 1990, and 2.2% for January through March 1991. Virology, 1993 Apr, 193(2), 621 - 30 Nucleotide sequence and genomic organization of rice tungro spherical virus; Shen P et al.; Rice tungro disease is caused by a combination of two viruses: rice tungro spherical virus (RTSV) and rice tungro bacilliform virus (Jones et al . (1991) J . Gen . Virol . 72, 757-761.) . The genome of RTSV is a single-stranded polyadenylated RNA . We present here the 12,433-nucleotide complete sequence of RTSV genomic RNA and its deduced coding regions . This sequence contains a large open reading frame (ORF) which initiates following a 514-nucleotide 5' leader sequence and is capable of encoding a viral polyprotein of 390.3 kDa . Two viral subgenomic RNAs of ca . 1.2 and 1.4 kb, respectively, were detected in RTSV-infected leaf tissues and mapped by S1 nuclease protection assay . These RNAs were determined to be congruent with the genomic RNA sequence proximal to the 3' terminus and could contain up to two small ORFs in their 5' to 3' orientation . There are at least three capsid protein subunit cistrons near the N-terminus of the large ORF . A computer-aided search of the C-terminal half of the large ORF revealed conserved protein sequence motifs for a viral RNA polymerase, proteinase, and a putative NTP-binding protein . These sequence motifs are arranged in a manner that resembles those of picorna-like viruses . Taken together, these data indicate that RTSV is a distinct type of positive-strand RNA virus . The evolutionary relationships between RTSV and other picorna-like plant viruses are discussed. J Urol, 1993 Apr, 149(4), 744 - 8 Intravesical bacillus Calmette-Guerin immunoprophylaxis of superficial bladder cancer: results of a controlled prospective trial with modified treatment schedule; Melekos MD et al.; A controlled prospective trial on 94 patients evaluated the efficacy of intravesical Pasteur strain bacillus Calmette-Guerin (BCG) administration as prophylaxis against tumor recurrences after complete endoscopic resection of superficial bladder cancer . The treatment schedule, consisting of an initial 6-week course of instillations and a single quarterly maintenance dose to the responders, was modified to those of the latter who were at high risk for recurrence and who received an additional separate 4-week course of therapy . The percentage of the patients treated prophylactically with BCG and who remained free of recurrences (68%, mean followup 33.8 months) was significantly higher than that of the controls who underwent transurethral resection only (41%, mean followup 30.2 months) . In terms of relative risk of recurrences, recurrence rate per 100 patient-months and disease-free interval, comparisons between the 2 groups of patients revealed a significant benefit for the BCG group overall as for those subjects having stages Ta and T1 tumors, multifocal tumors, a history of disease, and grades 2 and 3 carcinoma . Drug-induced toxicity was acceptable . Our study suggests that our modified treatment protocol is notably safe and effective against recurrent superficial bladder cancer. J Immunol, 1993 Apr 1, 150(7), 3101 - 7 Immunization with recombinant BCG-SIV elicits SIV-specific cytotoxic T lymphocytes in rhesus monkeys; Yasutomi Y et al.; Because the transmission of HIV is likely to occur through cell-associated virus, an effective HIV vaccine should be capable of eliciting HIV-specific CTL . We have employed the simian immunodeficiency virus (SIV)/rhesus monkey model to explore the use of the attenuated tuberculosis bacillus, Calmette Guerin bacillus (BCG), as a vaccine vehicle to elicit AIDS virus-specific CTL . BCG was engineered to express SIVmac gag under the control of hsp70 regulatory sequences . Immunization with this rBCG-SIVmac gag elicited MHC class I-restricted, CD8+ SIVmac gag-specific CTL in rhesus monkeys . In fact, SIVmac gag-specific CTL could be cloned readily from peripheral blood lymphocytes of these immunized monkeys . These findings provide further evidence for the power of BCG as a vaccine vector and its continued exploration as a vehicle for eliciting HIV-specific immunity. Zhonghua Bing Li Xue Za Zhi, 1993 Apr, 22(2), 80 - 2 {Localization and significance of lysozyme in tuberculosis}; Zhang SF; The relationship between expression of lysozyme and tuberculosis lesions was studied in 132 patients . The positive rate of lysozyme expression in 132 tuberculosis cases was 83.3% . Lysozyme was mainly distributed intracellularly, and the positive rates in different types, i.e . in cases with lesions of caseous necrosis, cellular nodes, and fibroid nodes were 90.1%, 79.3% and 63.3% respectively (P < 0.05 between lesions of caseous necrosis and fibroid nodes) . The positive rate of lysozyme in cases of infection associated with L-form tubercule bacillus was 64.4%, which was higher in cases with positive L-form antigen than that in antigen negative cases (P < 0.025) . The results indicated that expression of lysozyme was rather active, and might be considered as a marker for the prognosis of tuberculosis treatment. J Korean Med Sci, 1993 Apr, 8(2), 135 - 44 Analysis of the immunologic mechanism of intravesical bacillus Calmette-Guerin therapy for superficial bladder tumors: distribution and function of immune cells; Chung JY et al.; Intravesical bacillus Calmette-Guerin (BCG) administration has been used as an adjuvant therapy after transurethral resection for superficial bladder cancer, but the exact mechanisms of its antitumor activity are not yet known . The aim of this study was to characterize the immunologic aspects of antitumor activity of BCG using an animal model . C3H/He inbred mice and murine bladder tumor cell line, MBT-2 were used . The changes in immune cells such as helper T cells, suppressor T cells, macrophages and natural killer cells in the bladder and spleen were analysed by immunohistochemical method in intravesical BCG instilled in normal bladder, MBT-2 implanted after electrocauterization of the bladder mucosa and MBT-2 implanted and intravesical BCG treated group . The changes in natural killer cell activity of the splenocytes and peritoneal lymphocytes were evaluated using 51chromium release assay at regular time intervals following intraperitoneal BCG instillation . The prophylactic anticancer effect was evaluated by observing the tumor growth in the intravesically BCG treated group after intravesical MBT-2 implantation . In immunohistochemical examination, a remarkable infiltration of macrophage and helper T cell was observed in the lamina propria of the bladder, and the helper and suppressor T cells ratio (Th/Ts ratio) was increased after intravesical BCG therapy . In 51chromium release assay, enhanced natural killer cell activity of the splenocytes and peritoneal lymphocytes was observed after intraperitoneal BCG inoculation . The growth of implanted tumor was suppressed following intravesical instillation of BCG . These results suggest that the antitumor activity of BCG is not related to the simple inflammatory reaction but to the local and systemic immune response in which helper T lymphocytes and mononuclear cells play an important role. J Gen Microbiol, 1993 Apr, 139 ( Pt 4), 661 - 7 A partial physical map for the chromosome of alkalophilic Bacillus sp . strain C-125; Sutherland KJ et al.; Bacillus sp . strain C-125 has been chosen as a model alkalophilic bacterium to understand how adaptation to growth at high pH is achieved . To aid genetic analysis, we have started characterization of its genome . By using the two infrequently-cutting restriction endonucleases, AscI and Sse8387I, in conjunction with pulsed-field electrophoretic techniques, the size of the genome was found to be 3.7 Mb . Southern blot analysis of single, double and partial digests of Bacillus sp . strain C-125 DNA, using AscI-linking clones, gene probes and purified Bacillus sp . strain C-125 restriction fragments, allowed a putative chromosome map to be constructed. Int J Immunopharmacol, 1993 Apr, 15(3), 371 - 82 delta-9-Tetrahydrocannabinol inhibits cell contact-dependent cytotoxicity of Bacillus Calmétte-Guérin-activated macrophages; Burnette-Curley D et al.; The effect of delta-9-tetrahydrocannabinol (delta-9-THC), the major psychoactive component of marijuana, on the capacity of Bacillus Calmette-Guerin (BCG)-activated macrophages to lyse L929 tumor cells, Naegleria fowleri amoebae, and herpes simplex virus-infected cells was examined . Delta-9-THC inhibited tumoricidal and amoebicidal activity in a dose-related manner . Antiviral activity was decreased when mice received 25 and 50 mg/kg delta-9-THC . The cannabinoid did not directly suppress the activation of macrophages as determined by levels of 5'-nucleotidase activity and did not inhibit splenic T-lymphocytes of BCG-recipient mice from producing interferon gamma . Nomarski optics microscopy, scanning electron microscopy, and radiolabeling binding studies demonstrated that macrophages from delta-9-THC-treated mice retained their capacity to attach to their targets . These results suggest that delta-9-THC suppresses cell contact-dependent amoebicidal, tumoricidal, and antiviral activities of activated macrophages at a stage following effector cell-target cell conjugation. J Bacteriol, 1993 Apr, 175(7), 2137 - 42 Two glucose transport systems in Bacillus licheniformis; Tangney M et al.; Bacillus licheniformis NCIB 6346 showed active accumulation of glucose which was inhibited by agents which affect the transmembrane proton gradient . Phosphotransferase (PTS) activity, identified as phosphoenolpyruvate-dependent phosphorylation of glucose, was found in cell extracts but could not be demonstrated in cells permeabilized with toluene when assays were conducted at pH 6.6 . The same was true for mannitol and fructose phosphotransferase activities . Cells grown on fructose accumulated glucose at a slower rate than glucose-grown cells, and extracts prepared from them did not contain glucose PTS activity . Examination of the effects of analogs on glucose uptake and phosphorylation showed that 2-deoxyglucose was not a PTS substrate, but did markedly inhibit glucose uptake, with stronger inhibition in cells grown on fructose . Glucose accumulation by whole cells grown on glucose became less sensitive to the uncoupler tetrachlorosalicylanilide (TCS) as the pH was raised from 6.6 to 8.0, while in fructose-grown cells TCS was equally effective across this pH range . PTS activity was exhibited by toluene-treated cells at pH 7.5 and above, although the system itself in extracts was not affected by pH in the range of 5.0 to 8.0 . The results are consistent with the presence of two glucose transport systems, one a PTS and the other operating by an alternative mechanisms, and suggest that the PTS in B . licheniformis may be regulated in a pH-dependent manner. Wei Sheng Wu Xue Bao, 1993 Apr, 33(2), 115 - 21 {Purification and properties of beta-mannanases from alkalophilic Bacillus N16-5}; Tian X et al.; Three extracellular alkaline beta-mannanases from an alkalophilic Bacillus N16-5 were purified to electrophoretic homogeneity by (NH4)2SO4 precipitation, DEAE-Sephadex A-25 chromatography, hydroxyapatite chromatography and preparative electrophoresis . Molecular weights and pI values of the beta-mannanases (M-1, M-2 and M-3) were 51000, 38000 and 34700 by SDS-PAGE and 4.3, 2.5 and 2.5 by PAGEIEF, respectively . The optimum pH for enzyme catalysis were 9.0 for M-1 and 10.0 for M-2 and M-3, respectively . All of three enzymes were the most active at 70 degrees C . The activities of three enzymes were strongly inhibited by Ag1+, Hg2+ and Mn2+ . Michaelis constants (Km) of the enzyme M-1, M-2 and M-3 for mannase from konjak were 2.9, 1.7 and 12.5 mg/ml, maximum velocities (Vmax) for the saccharide were 27500, 47500 and 15700 mumol.min-1.mg-1, respectively . Konjak was hydrolyzed by the enzyme M-1, and major components in digests were mono-, di-, tri- and tetra-saccharides. Mol Ecol, 1993 Apr, 2(2), 65 - 78 Variable stability of antibiotic-resistance markers in Bacillus cereus UW85 in the soybean rhizosphere in the field; Halverson LJ et al.; We compared the stability of antibiotic-resistance markers in strains derived from Bacillus cereus UW85 in culture media and in the soybean rhizosphere in a growth chamber and in the field . We studied two independent, spontaneous mutants resistant to neomycin, three independent, spontaneous mutants resistant to streptomycin, and strains carrying plasmid pBC16, which encodes tetracycline resistance . Antibiotic-resistance markers were maintained in populations of all UW85 derivatives in culture and in the rhizosphere of soybeans grown in soil in a growth chamber . In two field experiments, antibiotic resistance was substantially lost in rhizosphere populations of B . cereus as early as 14 or as late as 116 days after planting . To distinguish between death of the inoculated strain and loss of its marker, we tested populations of B . cereus for other phenotypes (orange pigmentation, plasmid-borne resistance to tetracycline, and biocontrol activity) that are typical of UW85-derivatives used as inoculum, but atypical of the indigenous populations of B . cereus, and these phenotypes were maintained in populations from which the marker was lost . In general, neomycin-resistance markers were maintained at a higher frequency than streptomycin-resistance markers, and maintenance of antibiotic-resistance markers varied with position on the root and with the year of the experiment . In a semi-defined medium, the UW85 derivatives grew at the same rate as the wild type at 28 degrees C, but most grew more slowly than the wild type at 16 degrees C, demonstrating that antibiotic resistance can affect fitness under some conditions . The results suggest that the stability of antibiotic-resistance markers should be assessed in the ecosystems in which they will be studied. Chest, 1993 Apr, 103(4), 1087 - 90 Use of BCG vaccine in shelters for the homeless . A decision analysis; Nettleman MD; As a result of many interacting variables, including crowded shelters and limited access to health care, homeless persons are at high risk for tuberculosis . Using traditional approaches, control of tuberculosis in this population has been difficult . Decision analysis was used to investigate the cost-effectiveness of BCG (bacillus Calmette-Guerin) vaccination in persons attending homeless shelters . This vaccination was cost-effective over a wide range of assumptions . Using conservative assumptions, a vaccine that was at least 40 percent effective would result in a net cost savings . If the efficacy of the vaccine were 50 percent, $4,000 would be saved, 12 life-years gained, and 23 cases of active tuberculosis prevented for every 1,000 persons vaccinated . Further study of the BCG vaccine in homeless persons and other populations at risk is warranted. J Formos Med Assoc, 1993 Apr, 92(4), 317 - 23 Gram-negative bacillary meningitis in adults; Hsu GJ et al.; To evaluate the clinical aspects of gram-negative bacillary meningitis (GNBM), we reviewed 41 adult patients with bacteriologically proven gram-negative bacillary meningitis, seen from 1985 to 1990 . Thirty-two patients had post-neurosurgical GNBM and nine patients had spontaneous GNBM . Spontaneous GNBM appeared to have a sudden onset, a relatively fulminant course, and was caused most often by Escherichia coli . Post-neurosurgical GNBM, however, had a more insidious onset, a more protracted course, and was more often caused by nosocomial organisms which were resistant to multiple antibiotics . The overall mortality was 39% . Patients treated with combined aminoglycoside therapy had a lower mortality rate than those treated with intravenous aminoglycoside (17% vs 48%) . The use of third-generation cephalosporins has made a significant therapeutic advance in the treatment of GNBM, with a lower mortality of 21% . We recommend treatment of GNBM with third-generation cephalosporins and aminoglycosides . If aminoglycosides are to be employed, it is suggested that they be administered both intravenously and directly into the central nervous system. Tuber Lung Dis, 1993 Apr, 74(2), 96 - 9 DNA analysis demonstrates that mycococcus forms are not mycobacteria; de Wit D et al.; Previous experimental evidence has suggested that the mycobacteria may exist in morphological forms other than the well characterised acid-fast bacillus . However none of these studies have been able to show conclusively that these variant forms are derived from the mycobacteria and therefore much debate has centered around the exact nature of these organisms . In this study we have examined stored cultures of the mycococcus form of Mycobacterium bovis BCG and M . phlei which were prepared by Csillag in 1972 and 1969 . Restriction fragment patterns of the DNA of the variant forms and the parent mycobacteria were not similar and chromosomal DNA from the parent mycobacteria did not hybridise with the DNA of the variant forms . Furthermore biochemical studies indicate that the variant forms are environmental contaminants . Although this study shows conclusively that the mycococcus is not derived from the mycobacteria, we believe that the search for variant morphological or metabolic forms of the mycobacteria should remain an active area of investigation, but that no study should be considered complete without the full use of newer molecular technology. J Bacteriol, 1993 Apr, 175(8), 2314 - 20 Cytotoxicity and ADP-ribosylating activity of the mosquitocidal toxin from Bacillus sphaericus SSII-1: possible roles of the 27- and 70-kilodalton peptides; Thanabalu T et al.; Clones expressing regions of the 100-kDa Bacillus sphaericus SSII-1 mosquitocidal toxin (Mtx) as fusion proteins with glutathione S-transferase were constructed, and the toxin-derived peptides were purified . The in vitro ADP-ribosylation activities of these peptides and their effects on larvae and cells in culture were studied . Mtx25 (amino acids 30 to 493) was found to ADP-ribosylate two proteins with molecular masses of 38 and 42 kDa, respectively, in Culex quinquefasciatus (G7) cell extracts, in addition to ADP-ribosylating itself . Mtx21 (amino acids 30 to 870; or a combination of Mtx25 and Mtx26 (amino acids 259 to 870) caused mortality in C . quinquefasciatus larvae . Mtx25, Mtx26, or Mtx24 (amino acids 30 to 276) alone and Mtx24 in combination with Mtx26 were not toxic to larvae . Mtx21 and Mtx26 produced marked morphological changes in G7 cells and to a lesser extent in Aedes aegypti cells but had no effect on Anopheles gambiae or HeLa cells . Thus, a domain in the N-terminal region of the Mtx protein is sufficient for ADP-ribosylation of C . quinquefasciatus cell protein, and a domain in the C-terminal region is sufficient for toxicity to cultured C . quinquefasciatus cells; however, both regions are necessary for toxicity to mosquito larvae. Biosci Biotechnol Biochem, 1993 Apr, 57(4), 584 - 90 Importance of the central region of 130-kDa insecticidal proteins of Bacillus thuringiensis var . israelensis for their activity in vivo and in vitro; Yoshida K et al.; To delineate the mosquitocidal regions of the ISRH3 (CryIVB) and ISRH4 (CryIVA) proteins, which are two of the mosquitocidal 130-kDa proteins contained in the crystalline protein bodies (CPBs) of Bacillus thuringiensis var . israelensis (BTI), a deletion analysis of these protein genes has been done . Based on the evidence that each 130-kDa protein had two mosquitocidal regions, N-terminal and C-terminal ones, and these two regions shared a common part in the center of the 130-kDa proteins, deleted genes on this region were constructed . As the protein products which lacked the central region had reduced activities, the central region could be important for the mosquitocidal activity . The mosquitocidal and non-mosquitocidal truncated gene products of 130-kDa protein genes were also applied to a cultured lepidopteran cell line, TN-368 . The mosquitocidal proteins caused the swelling and disruption of the cells in spite of the insecticidal specificity of CPBs of BTI, but the non-mosquitocidal proteins did not . Therefore, TN-368 cells were sensitive to the mosquitocidal fragments of 130-kDa proteins of BTI under the assay conditions used. Appl Microbiol Biotechnol, 1993 Apr, 39(1), 63 - 8 Production and immobilization of a proteinase-reduced cyclodextrin glycosyltransferase preparation; Steighardt J et al.; Cyclodextrin glycosyltransferase (CGTase) was produced by a 3-day cultivation of Bacillus macerans growing in a natural medium containing grated-potatoes . Besides CGTase the culture supernatant contained a mixture of serine proteinases with a predominant subtilisin-like activity . By fractionated precipitation with ammonium sulphate the CGTase (molecular mass approximately 70 kDa) was concentrated and largely separated from the proteinases (molecular mass approximately 28 kDa) . Among the various immobilization methods and carrier materials tested the enriched CGTase was covalently bound preferably onto porous glass beads using glutardialdehyde as cross-linker . The discontinuous conversion of soluble starch into beta-cyclodextrin was carried out with native as well as immobilized CGTase over 24 h . The batch re-usability of the fixed enzyme proved to be at least 20 times with a residual CGTase activity of 65%. Immunology, 1993 Apr, 78(4), 635 - 42 Induction of an auto-anti-IgE response in rats . IV . Effects on mast cell degranulation; Jaffery G et al.; Induction of an auto-anti-IgE (auto-aIgE) response in the rat inhibits both total and specific IgE production and alters the distribution of mast cell (MC) subpopulations identified by differential Alcian blue/safranin staining . We have extended these observations by characterizing the auto-aIgE antibodies and determining their effects on MC degranulation in vitro and in vivo . An auto-aIgE response was induced in bacillus Calmette-Guerin (BCG)-primed rats by injecting a conjugate of highly purified rat IgE myeloma (IR2) coupled to tuberculin-derived purified protein derivative (PPD) . Anti-IgE autoantibodies were almost exclusively IgG2a . The intradermal injection of auto-aIgE into untreated rats induced local MC degranulation as shown by a strong immediate skin response . Histologically there was evidence of significant degranulation of safranin staining connective tissue MC (SMC) in the skin but not of the Alcian blue staining MC (ABMC) in the sub-epidermal region . The induced degranulation was epsilon-chain specific; immunopurified anti-idiotypic antibodies raised to the IgE IR2 myeloma had no MC degranulating activity . When administered locally, auto-aIgE inhibited a subsequent passive cutaneous anaphylaxis (PCA) response elicited by anti-ovalbumin IgE . In addition, the PCA response was significantly decreased in animals with an ongoing auto-aIgE response . Immunopurified auto-aIgE also induced histamine release in vitro from rat peritoneal MC . These results are discussed in the context of naturally occurring autoantibodies to IgE present in patients with allergic disease. Biochem Biophys Res Commun, 1993 Mar 31, 191(3), 922 - 7 Specific interaction of guanidine hydrochloride with essential carboxyl group of xylanase from alkalothermophilic Bacillus sp; Chauthaiwale J et al.; Experimental evidence for the specific interaction of guanidine hydrochloride with the carboxyl group of xylanase has been presented for the first time . Guanidine hydrochloride (0.1 M) inactivated the xylanase from alkalothermophilic Bacillus sp . to 50% without affecting the conformation of the protein as determined by the fluorometric profile . The kinetic analysis indicated a competitive type of inhibition and a requirement of 1.4 molecules of guanidine hydrochloride per molecule of the enzyme for inhibition . Maximum inhibition occurred at the pH which is optimum for the enzyme activity . The reaction of guanidine hydrochloride with the enzyme prior to modification by Woodward's Reagent K, a specific inhibitor of the carboxyl group, made it inaccessible for modification as indicated by absorbance data at 340 nm . Urea, sodium dodecyl sulphate, LiCl, KCl and NaCl at 0.1 M concentration each had negligible effect on the enzyme activity. Biochemistry, 1993 Mar 23, 32(11), 2845 - 52 Significance of hydrophobic S4-P4 interactions in subtilisin 309 from Bacillus lentus; Bech LM et al.; The subtilisins have an extended substrate binding cleft comprising at least 8 subsites . Two pockets at the S1 and S4 sites are particularly conspicuous, and the interactions between substrate and these two pockets are very important for the substrate specificity . Phe residues have mutationally been introduced at one of positions 102, 128, 130, and 132 of the subtilisin Savinase from Bacillus lentus to investigate the effects of introducing bulky groups along the rim of the S4 binding pocket . It is shown that the marked P4 preference of wild-type Savinase for aromatic groups is eliminated by the Gly102-->Phe and Ser128-->Phe mutations, indicating that bulky groups at positions 102 and 128 block the S4 binding site . In contrast, the activity toward hydrophilic P4 residues is not nearly as affected by these mutations, suggesting that the binding mode of the P4 side chain is dependent on its properties . Introduction of a bulky -CH2-S-CH2-CH2-pyridyl group at position 128, by mutational incorporation of Cys followed by chemical modification with 2-vinylpyridine, has essentially the same effect . The Ser130-->Phe mutation hardly affects the activity of the enzyme while the Ser-->Phe mutation at position 132 renders the preference for hydrophobic groups in P4 even more pronounced . This mutation furthermore affects the size of the S4 pocket . An analysis of double mutants at positions 132 and 104 suggests that the S4 region is flexible and is adjusted upon binding of substrates. Eur J Biochem, 1993 Mar 15, 212(3), 801 - 9 Structural studies of the O-antigenic polysaccharide of Fusobacterium necrophorum; Hermansson K et al.; The O-specific polysaccharide component of the lipopolysaccharide produced by Fusobacterium necrophorum is of the teichoic acid type, with repeating units connected by phosphoric diester linkages . Dephosphorylation of the polysaccharide by treatment with aqueous hydrogen fluoride yielded a carbohydrate composed of a trisaccharide linked to an acidic component . This product, and the polysaccharide, were investigated by chemical methods and 1H-, 13C-, 31P- and 15N-NMR spectroscopy and the former also by fast-atom-bombardment mass spectrometry . It is proposed that the polysaccharide is composed of repeating units having the following structure, in which Fuc represents fucose (6-deoxy-galactose), Am represents an acetamidino group and Sug 2,4-diamino-2,4,6-trideoxy-D-glucose ('bacillosamine') acetylated at the 2-position and acylated with a (S)-3-hydroxybutanoic acid at the 4-position . The acid was identified as a 2-amino-2-deoxy-2-C-methyl-pentonic acid (2-amino-2-methyl-3,4,5-trihydroxypentanoic acid) . The configuration of this acid remains to be determined . {formula: see text} Proc Natl Acad Sci U S A, 1993 Mar 15, 90(6), 2355 - 9 Structure of glycosomal glyceraldehyde-3-phosphate dehydrogenase from Trypanosoma brucei determined from Laue data; Vellieux FM et al.; The three-dimensional structure of glycosomal glyceraldehyde-3-phosphate dehydrogenase {D-glyceraldehyde-3-phosphate:NAD+ oxidoreductase (phosphorylating), EC 1.12.1.12} from the sleeping-sickness parasite Trypanosoma brucei was solved by molecular replacement at 3.2-A resolution with an x-ray data set collected by the Laue method . For data collection, three crystals were exposed to the polychromatic synchrotron x-ray beam for a total of 20.5 sec . The structure was solved by using the Bacillus stearothermophilus enzyme model {Skarzynski, T., Moody, P . C . E . & Wonacott, A . J . (1987) J . Mol . Biol . 193, 171-187} with a partial data set which was 37% complete . The crystals contain six subunits per asymmetric unit, which allowed us to overcome the absence of > 60% of the reflections by 6-fold density averaging . After molecular dynamics refinement, the current molecular model has an R factor of 17.6% . Comparing the structure of the trypanosome enzyme with that of the homologous human muscle enzyme, which was determined at 2.4-A resolution, reveals important structural differences in the NAD binding region . These are of great interest for the design of specific inhibitors of the parasite enzyme. Brain Res, 1993 Mar 5, 605(1), 155 - 63 Membrane-bound choline-O-acetyltransferase in rat hippocampal tissue is anchored by glycosyl-phosphatidylinositol; Smith LK et al.; In an earlier study, we presented evidence to suggest that some of the particulate choline-O-acetyltransferase (ChAT) in rat hippocampal tissue might be linked to membranes by a glycosyl-phosphatidylinositol (GPI) anchor . In the present report, we attempted to determine if any of this GPI-anchored ChAT might be intracellular . Internalization of phosphatidylinositol-specific phospholipase C (PI-PLC) from Bacillus thuringiensis into rat hippocampal synaptosomes by the DMSO (dimethyl sulfoxide) freeze/thawing procedure caused an increase in cytosolic and a decrease in membrane-bound ChAT . Incubation of a plasma membrane enriched subcellular fraction at 16 degrees C relative to 4 degrees C led to a conversion of the membrane-bound, amphiphilic ChAT into hydrophilic ChAT . This conversion was blocked by zinc, an inhibitor of GPI-PLC . The cytosolic fraction of ChAT immunoreacted on western blots with an antibody directed against the cross-reacting determinant (CRD) of the GPI anchor . We suggest that some of the membrane-bound ChAT in rat hippocampal tissue is GPI-anchored intracellularly; also, that an endogenous GPI-PLC-like enzyme acts to release it into the cytosol. J Mol Biol, 1993 Mar 5, 230(1), 323 - 41 The high-resolution structure of the peripheral subunit-binding domain of dihydrolipoamide acetyltransferase from the pyruvate dehydrogenase multienzyme complex of Bacillus stearothermophilus; Kalia YN et al.; The three-dimensional structure of a 43-residue active, synthetic peptide encompassing the peripheral subunit-binding domain of dihydrolipoamide acetyltransferase from the pyruvate dehydrogenase multienzyme complex of Bacillus stearothermophilus has been determined by means of a multi-cooling dynamical simulated annealing protocol using restraints derived from 1H nuclear magnetic resonance spectroscopy . A total of 442 experimentally derived restraints including 13 dihedral angle (phi, chi 1) restraints were used . A final set of 35 structures was calculated with a root-mean-square deviation from the mean co-ordinates of 0.36 A for the backbone atoms and 0.96 A when side-chain heavy atoms were included for the well-defined region comprising residues Val7 to Leu39 . Although assignments were made and sequential connectivities observed for the N-terminal six and C-terminal four residues, the absence of long-range NOEs suggests that the terminal regions are largely unstructured . The binding domain contains two short parallel alpha-helices (residues Val7 to Lys14 and Lys32 to Leu39), a3(10)-helix (residues Asp17 to Val21) and a structured loop made up of overlapping beta-turns (residues Gln22 to Leu31), which enclose a close-packed hydrophobic core . The loop is stabilized to a large extent by Asp34 . This residue is conserved in all peripheral subunit-binding domains and its carboxylate side-chain forms a set of side-chain-main-chain hydrogen bonds with the main-chain amide protons of Gly23, Thr24, Gly25 and Leu31 and a side-chain-side-chain hydrogen bond with the hydroxyl group of Thr24 . We propose that a peripheral subunit-binding site may be located in the loop region, which contains a series of highly conserved residues and provides a number of potential recognition sites . The structured region of the binding domain, comprising 33 residues, represents an exceptionally short amino acid sequence with defined tertiary structure that has no disulphide bond, ligand or cofactor to stabilize the fold . It may be approaching the lower size limit for a three-dimensional structure possessing features characteristic of larger structures, including a close-packed, non-polar interior . The organization of the side-chains in the hydrophobic core may have implications for de novo protein design. Yakugaku Zasshi, 1993 Mar, 113(3), 264 - 71 {Inducible resistance to macrolide-lincosamide-streptogramin type B antibiotics in Bacillus licheniformis: common structures of macrolide antibiotics capable of inducing the resistance}; Kobayashi H et al.; Whether or not resistance to macrolide-lincosamide-streptogramin type B antibiotics (MLS) can be induced by many macrolide antibiotics (Mac), was inquired in Bacillus licheniformis EMR . Resistance to MLS in the strain was induced by erythromycin, oleandomycin, clarithromycin, roxithromycin, narbomycin, picromycin, kujimycin A or B, mycinamicin I, or rosamicin . On the contrary, josamycin, spiramycin, tylosin, rokitamycin, midecamycin, and miokamycin as well as lincosamide and streptogramin type B antibiotics could not induce MLS-resistance . The results suggest that two common chemical residues of the inducer Mac, that is, 1) a single monosaccharide at C5 in the 14- and 16-membered lactone rings, and 2) one polar group such as dimethylamino or methoxyl at C3' in the sugar, are likely to be responsible for showing the activity of MLS-resistance inducer in Bacillus licheniformis EMR. Plant Mol Biol, 1993 Mar, 21(6), 1131 - 45 The reconstruction and expression of a Bacillus thuringiensis cryIIIA gene in protoplasts and potato plants; Adang MJ et al.; A Bacillus thuringiensis (B.t.) cryIIIA delta-endotoxin gene was designed for optimal expression in plants . The modified cry gene has the codon usage pattern of an average dicot gene and does not contain AT-rich nucleotide sequences typical of native B.t . cry genes . We assembled the 1.8 kb cryIIIA gene in nine blocks of three oligonucleotide pairs . For two DNA blocks, the polymerase chain reaction was used to enrich for correctly ligated pairs . We compared modified cryIIIA gene with native gene expression by electroporation of dicot (carrot) and monocot (corn) protoplasts . CryIIIA-specific RNA and protein was detected in carrot and corn protoplasts only after electroporation with the rebuilt gene . Transgenic potato lines were generated containing the redesigned cryIIIA gene under the transcriptional control of a chimeric CaMV 35S/mannopine synthetase (Mac) promoter . Out of 63 transgenic potato lines, 58 controlled first-instar Colorado potato beetle (CPB) larvae in bioassays . Egg masses which produced ca . 250,000 CPB larvae were placed on replicate clones of 56 transgenic potatoes . No CPB larvae developed past the second instar on any of these plants . Plants expressing high levels of delta-endotoxin were identified by their toxicity to more resistant third-instar larvae . We show there was good correlation between insect control and the levels of delta-endotoxin RNA and protein. Mol Biol (Mosk), 1993 Mar-Apr, 27(2), 416 - 28 {Comparison of the heat stability and structure close homologs--Bacillus amyloliquefaciens ribonuclease and Bacillus intermedius 7P ribonuclease}; Makarov AA et al.; Parameters of heat denaturation and intrinsic fluorescence of barnase and its close homologue, binase, in the pH region 2-6 have been determined . Barnase heat denaturation (pH 2.8-5.5) proceeds according to the "all-or-none" principle . Barnase denaturation temperature is lower than that of binase and this difference increases from 2.5 degrees C at pH 5 to 7 degrees C at pH 3 . Enthalpy values of barnase and binase denaturation coincide only at pH 4.5-5.5, but as the pH decreases the barnase denaturation enthalpy decreases significantly and in this respect it differs from binase . The fluorescence and CD techniques do not reveal any distinctions in the local environment of aromatic residues in the two proteins, and the obtained difference in the parameters of intrinsic fluorescence is due to fluorescence quenching of the barnase Trp-94 by the His-18 residue, which is absent in binase . Secondary structures of both native and denaturated proteins also do not differ . Some differences have been found in the barnase and binase electrostatic characteristics, revealed in the character of the dipole moment distribution. J Biochem (Tokyo), 1993 Mar, 113(3), 355 - 63 Thermostable farnesyl diphosphate synthase of Bacillus stearothermophilus: molecular cloning, sequence determination, overproduction, and purification; Koyama T et al.; The structural gene for thermostable farnesyl diphosphate synthase from Bacillus stearothermophilus was cloned, sequenced, and overexpressed in Escherichia coli cells . A 1,260-nucleotide sequence of the cloned fragment was determined . This sequence specifies an open reading frame of 891 nucleotides for farnesyl diphosphate synthase . The deduced amino acid sequence shows a 42% similarity with that of E . coli FPP synthase {Fujisaki et al . (1990) J . Biochem . 108, 995-1000} . Comparison with prenyltransferases from a wide range of organisms, from bacteria to human, revealed the presence of seven highly conserved regions . In contrast to thermolabile prenyltransferases, which have four to six cysteine residues, the thermostable farnesyl diphosphate synthase carries only two cysteine residues . This enzyme is also unique in that some of the amino acids that are fully conserved in equivalents from other sources are replaced by functionally different amino acids . Construction of an overproducing strain provided a sufficient supply of this enzyme and it was purified to homogeneity . The purified recombinant enzyme is immunochemically identical with the native B . stearothermophilus enzyme, and it is not inactivated even after treatment at 65 degrees C for 70 min. Insect Biochem Mol Biol, 1993 Mar, 23(2), 273 - 83 In vitro and in vivo proteolysis of the Bacillus thuringiensis subsp . israelensis CryIVD protein by Culex quinquefasciatus larval midgut proteases; Dai SM et al.; Proteases with trypsin-, chymotrypsin- and thermolysin-like specificity were detected in Culex quinquefasciatus larval midguts . Their activities were monitored by N-terminal amino acid sequence analysis of the Bacillus thuringiensis subsp . israelensis CryIVD toxin proteolytic fragments . These proteases are located in the larval midgut and in different fractions obtained during the preparation of brush border membrane vesicles . The activity of the midgut proteases increased with an increase in pH . Both the chymotrypsin- and thermolysin-like activities are involved in the processing of solubilized CryIVD toxin, whereas an additional trypsin-like protease is necessary for the CryIVD parasporal inclusion processing . The solubilized CryIVD toxin was first cleaved between Thr347 and Phe348 and between Phe348 and Tyr349, generating a 40-kDa N-terminal fragment and a 32.5-kDa C-terminal fragment . The C-terminal domain was resistant to further processing, with only a small amount of a 31-kDa product appearing due to the action of a thermolysin-like protease . However, the N-terminal domain was very unstable, and was further degraded to about 30 kDa . Unlike the solubilized CryIVD toxin, the processing of the CryIVD parasporal inclusion was very slow at neutral pH . Three protease-resistant products were detected at pHs higher than 9.5 with an overnight incubation at 37 degrees C . The 30- and 28.5-kDa C-terminal peptides are proteolytic products of trypsin- and chymotrypsin-like proteases, respectively; while the 28-kDa N-terminal peptide has 27 amino acids deleted from the N-terminal end by a thermolysin-like protease. Biokhimiia, 1993 Mar, 58(3), 340 - 7 {Characteristics of phospholipid hydrolysis kinetics by phospholipase C from Bacillus cereus . Hydrolysis of phosphatidylinositol in various aggregated states}; Selishcheva AA et al.; The process of phospholipase C hydrolysis of phosphatidylethanolamine (PE) in the form of mixed micelles phospholipid-detergent or in the form of vesicules in the mixture with phosphatidylcholine (PC) was studied . The size of the micelles was measured by dynamic light-scattering and their structure was determined by 31P-NMR spectroscopy . It was found that the kinetics of PE hydrolysis in the micelles by phospholipase C from Bacillus cereus do not follow Michaelis-Menten equation, but at all concentrations studied PE was hydrolyzed significantly slower than PC . The rate of PC hydrolysis was measured in the previous work . The hydrolysis of PE in PC-vesicules was followed with use of Victoria blue R dye method . It was shown that the rate of PE hydrolysis in the vesicules is similar to that of PC. Clin Exp Dermatol, 1993 Mar, 18(2), 133 - 7 Bacillary epithelioid angiomatosis in acquired immunodeficiency syndrome (AIDS)--clinicopathological and ultrastructural study of a case with a review of the literature; Innocenzi D et al.; Bacillary epithelioid angiomatosis (BEA) is a rare cutaneous disorder usually affecting patients with human immunodeficiency virus (HIV) infection often misdiagnosed as a vascular tumour . We describe a 51-year-old, HIV-positive, Caucasian, homosexual male who developed scattered papulo-nodular lesions with clinicopathological and ultrastructural features of BEA . He had a dramatic therapeutic response to systemic antibiotics . There has been a lack of such reports in the European literature . The differential diagnosis is discussed and a brief review of the English literature to date is included. Appl Environ Microbiol, 1993 Mar, 59(3), 927 - 32 Purification, characterization, gene cloning, and sequencing of a new beta-glucosidase from Bacillus circulans subsp . alkalophilus; Paavilainen S et al.; An intracellular beta-glucosidase was purified from cell extracts of Bacillus circulans subsp . alkalophilus by NAD affinity and high-performance anion-exchange chromatographies . The enzyme was active against a wide range of aryl-beta-glucosides and beta-linked disaccharides . The structural gene for beta-glucosidase was cloned in Escherichia coli . The beta-glucosidase gene consisted of an open reading frame of 1,350 bp encoding a protein of 450 amino acids with a calculated M(r) of 51,303 . The enzyme exhibited from 45 to 66% identity with five bacterial beta-glucosidases. Appl Environ Microbiol, 1993 Mar, 59(3), 815 - 21 High-level cryIVD and cytA gene expression in Bacillus thuringiensis does not require the 20-kilodalton protein, and the coexpressed gene products are synergistic in their toxicity to mosquitoes; Chang C et al.; Interactions among the 20-kDa protein gene and the cytA and cryIVD genes located in a 9.4-kb HindIII fragment were studied . A series of plasmids containing a combination of these different genes was constructed by using the Escherichia coli/Bacillus thuringiensis shuttle vector pHT3101 . The plasmids were then used to transform an acrystalliferous strain, cryB, derived from B . thuringiensis subsp . kurstaki . The results from sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot analyses suggest that although the 20-kDa protein is required for the efficient CytA protein production in E . coli, it is not required in B . thuringiensis . With or without the truncated 20-kDa protein gene, the CtyA and/or CryIVD proteins are produced and form parasporal inclusions in B . thuringiensis cells . However, more-efficient expression is obtained when a second protein, probably acting as a chaperonin, is present . In addition, the time course studies show that the CytA and CryIVD proteins are coordinately produced . Both the crude B . thuringiensis culture and purified inclusions from each recombinant B . thuringiensis strain are toxic to Culex quinquefasciatus larvae . The parasporal inclusions formed in B . thuringiensis cells are mosquitocidal, with CytA synergizing CryIVD toxicity. Gut, 1993 Mar, 34(3), 371 - 4 Antibodies to Mycobacterium paratuberculosis and nine species of environmental mycobacteria in Crohn's disease and control subjects; Stainsby KJ et al.; Cultural and serological studies have provided limited, often conflicting, evidence of a role for mycobacteria in the pathogenesis of Crohn's disease . Interest has focussed on Mycobacterium paratuberculosis, previously considered to be common in the environment with no major role as a human pathogen . Whether a specific serum antibody response to mycobacteria occurs in Crohn's disease or ulcerative colitis was investigated . Sera from patients with Crohn's disease (n = 38), ulcerative colitis (n = 15), and a healthy control population (n = 30) were assayed in an enzyme linked immunosorbent assay (ELISA) using eight filtered sonicate mycobacterial preparations and a purified protein derivative made from the bovine tubercle bacillus . In addition, IgG, IgM, and IgA levels to M paratuberculosis were determined in sera from patients with active (n = 24) or inactive (n = 29) Crohn's disease and the control populations . There was strong evidence of contact with environmental mycobacteria in all patients and control populations, with the greatest responses to preparations of M avium, M tuberculosis, and M kansasii . A large proportion of patients with Crohn's disease had antibodies that bound most antigens tested but there were no statistical differences between these values and those of the control population . Similarly, there were no differences in antibody levels to M paratuberculosis in patient and control groups . Although a subset of patients with active Crohn's disease (25%) had IgG concentrations that exceeded the control mean by more than 2 SD, this phenomenon may not be specific to Crohn's disease: 20% of a small group of patients with coeliac disease had similarly raised IgG levels to M paratuberculosis . These findings do not provide serological evidence of a role for this organism in the pathogenesis of Crohn's disease. FEMS Microbiol Lett, 1993 Mar 1, 107(2-3), 179 - 83 Analysis of common antigen of flagella in Bacillus cereus and Bacillus thuringiensis; Murakami T et al.; Flagellar antigen of Bacillus cereus H.1 was purified and tested for serodiagnostic antigen by ELISA . The antibody against the flagellar antigen of B . cereus H.1 reacted not only with the homologous specific antigen but also reacted with the flagellar antigens of 23 strains of B . cereus . This common flagellar antigen of B . cereus was found to be due to 61-kDa protein by SDS-PAGE and immunoblot assay . Monoclonal antibody H15A5 against common antigenic epitope of B . cereus also reacted with flagellar antigens of 21 strains of Bacillus thuringiensis by ELISA . This monoclonal antibody reacted with the 61-kDa protein of the flagella of B . cereus H.1 and H.2 and B . thuringiensis Kurstaki HD1, Alesti and Aizawai juroi by immunoblot analysis . These results indicated that the common antigenic epitope of the 61-kDa protein existed in the flagella both of B . cereus and B . thuringiensis. Trop Geogr Med, 1993 Mar, 45(1), 2 - 5 The approach to the leprosy problem in the past in Indonesia . A historical review of leprosy control activities in Indonesia during the last centuries; Zuiderhoek B; Due to the severe disabilities leprosy has always been a disease which appealed to the imagination . In bygone centuries cause and treatment were unknown and the fear to be infected was enormous . In Indonesia medical officers struggled with the problem . As early as the 17th century the disease was described in detail by Ten Rhijne . In the 19th century leprosy was considered hereditary . After the discovery of the leprosy bacillus by Hansen in 1873, confusion continued as the bacillus could not be cultivated . Many therapies were tried, but with no result . At first patients were isolated in leprosaria, later on a more humane system of house-isolation was introduced . Since 1932 Indonesian medical officers have played a prominent part in research and the determination of the future approach to the problem . Seen against the background of our present knowledge about the disease, it is interesting to follow the struggle against leprosy in the past. J Bacteriol, 1993 Mar, 175(6), 1814 - 22 Electron microscopic analysis and structural characterization of novel NADP(H)-containing methanol: N,N'-dimethyl-4-nitrosoaniline oxidoreductases from the gram-positive methylotrophic bacteria Amycolatopsis methanolica and Mycobacterium gastri MB19; Bystrykh LV et al.; The quaternary protein structure of two methanol:N,N'-dimethyl-4-nitrosoaniline (NDMA) oxidoreductases purified from Amycolatopsis methanolica and Mycobacterium gastri MB19 was analyzed by electron microscopy and image processing . The enzymes are decameric proteins (displaying fivefold symmetry) with estimated molecular masses of 490 to 500 kDa based on their subunit molecular masses of 49 to 50 kDa . Both methanol:NDMA oxidoreductases possess a tightly but noncovalently bound NADP(H) cofactor at an NADPH-to-subunit molar ratio of 0.7 . These cofactors are redox active toward alcohol and aldehyde substrates . Both enzymes contain significant amounts of Zn2+ and Mg2+ ions . The primary amino acid sequences of the A . methanolica and M . gastri MB19 methanol:NDMA oxidoreductases share a high degree of identity, as indicated by N-terminal sequence analysis (63% identity among the first 27 N-terminal amino acids), internal peptide sequence analysis, and overall amino acid composition . The amino acid sequence analysis also revealed significant similarity to a decameric methanol dehydrogenase of Bacillus methanolicus C1. Chest, 1993 Mar, 103(3), 839 - 43 Empyema of the thorax in adults . Etiology, microbiologic findings, and management; Alfageme I et al.; The etiology, microbiologic findings, and management of 82 episodes of empyema treated by our unit over a period of 6 years were analyzed . Average patient age was 54 years . Eighty-two percent had underlying disease such as alcoholism (29 percent), malignancy (23 percent), and diabetes mellitus (20 percent) . Sixty (73 percent) had an empyema develop secondary to a bronchopulmonary infection . Other etiologies were as follows: infradiaphragmatic sepsis, five cases; iatrogenic, ten cases; and idiopathic, seven cases . Cultures were positive in 76 cases and negative in the remaining 6 (2 positive Gram stains, 1 positive under bacilloscopy, and 3 were sterile) . Anaerobes were isolated from 25 and aerobes from 47 of the positive cultures . A single bacteria was isolated from 43 and multiple organisms (average: 2.63/case) grew on the remaining 33 positive cultures . Length of hospitalization averaged 37 days . Seven patients received antibiotics only, thoracentesis was performed on three, intercostal chest tube drainage was required in 72, and more aggressive surgery was performed on 12 patients (7 with fibrothorax and 5 with pneumonectomy) . Streptokinase was instilled into the pleural space of eight patients with good results . Pleural drainage superinfection occurred at a rate of 8.5 percent . Nine patients died; the remaining recovered . Only three deaths came about as a direct result of the empyema. Cancer, 1993 Mar 1, 71(5), 1846 - 7 Pulmonary granulomata . A complication of intravesical administration of bacillus Calmette-Guerin for superficial bladder carcinoma; Smith RL et al.; Intravesical administration of bacillus Calmette-Guerin (BCG) is an effective treatment for superficial carcinoma of the bladder . The authors report a pulmonary complication characterized by miliary infiltration on chest roentgenogram and caseating granulomata on lung biopsy specimens . This case and three prior reports suggest that this complication may be a form of hypersensitivity rather than true BCG infection. Am J Clin Pathol, 1993 Mar, 99(3), 244 - 8 Granulomatous inflammation in bladder wash specimens after intravesical bacillus