Nucleic Acids Res, 1985 Nov 25, 13(22), 8207 - 17 Molecular cloning and the nucleotide sequence of the Mr 28 000 crystal protein gene of Bacillus thuringiensis subsp . israelensis; Waalwijk C et al.; The Mr 28.000 crystal protein gene of Bacillus thuringiensis subspecies israelensis has been cloned into pBR322 as part of a 9.7 kb HindIII fragment . From hybridization experiments of recombinant p425 DNA with B.t . subspecies israelensis RNA from different stages of growth it was concluded that transcription of the gene is restricted to early sporulation stages . Nucleotide sequence analysis revealed the presence of a large open reading frame with a coding capacity of 249 amino acids (Mr 27.340) . Nuclease S1 mapping demonstrated that transcription starts 44 nucleotides upstream of the initiation codon . A Shine-Dalgarno sequence (AAGGAG) was found 10 nucleotides upstream of the translation startpoint . At the 3'-end of the gene a complex secondary structure was found immediately after the stop-codon . Despite the presence of these regulation signals only limited expression in E . coli was detected . This can be explained by assuming that B.t . subsp . israelensis promotor sequences are poorly recognized by E . coli RNA polymerase. Biochemistry, 1985 Nov 19, 24(24), 6876 - 87 Cryoenzymology of Bacillus cereus beta-lactamase II; Bicknell R et al.; The effects of cryosolvents and subzero temperatures on the metalloenzyme beta-lactamase II from Bacillus cereus have been investigated . Preliminary experiments led to the selection of suitable systems for the study of beta-lactamase II catalysis at low temperatures, namely, cobalt(II) beta-lactamase II hydrolysis of benzylpenicillin in 60% (v/v) ethylene glycol and zinc beta-lactamase II hydrolysis of the chromophoric cephalosporin nitrocefin in 60% (v/v) methanol . Progress curves for the hydrolysis of benzylpenicillin by cobalt beta-lactamase II in 60% (v/v) ethylene glycol at temperatures below -30 degrees C consisted of a transient followed by a steady-state phase . The amplitude of the transient implied a burst whose magnitude was greater than the concentration of enzyme, and the proposed mechanism comprises a branched pathway . The kinetics for the simplest variants of such pathways have been worked out, and the rate constants (and activation parameters) for the individual steps have been determined . The spectrum of the enzyme changed during turnover: when benzylpenicillin was added to cobalt beta-lactamase II, there was a large increase in the cysteine-cobalt(II) charge-transfer absorbance at 333 nm . This increase occurred within the time of mixing, even at -50 degrees C . The subsequent decrease in A333 was characterized by a rate constant that had the same value as the "branching" rate constant of the branched-pathway mechanism . This step is believed to be a change in conformation of the enzyme-substrate complex . Single-turnover experiments utilized the change in A333, and the results were consistent with pre-steady-state and steady-state experiments . When a single-turnover experiment at -48 degrees C was quenched with acid, the low molecular weight component of the intermediate was shown to be substrate . The mechanism advanced for the hydrolysis of benzylpenicillin by cobalt beta-lactamase II involves two noncovalent enzyme-substrate complexes that have been characterized by their electronic absorption spectra . When manganese beta-lactamase II was used, the same features (implying a branched pathway) were evident; these experiments were carried out at ordinary temperatures and did not utilize a cryosolvent . The hydrolysis of nitrocefin by zinc beta-lactamase II has been studied concurrently in 60% (v/v) methanol . Progress curves were triphasic . There were two transients preceding the linear steady-state phase . The stoichiometry of the burst again implied a branched pathway.(ABSTRACT TRUNCATED AT 400 WORDS) J Am Vet Med Assoc, 1985 Nov 15, 187(10), 1047 - 8 Bovine abortion caused by Bacillus cereus; Schuh J et al.; Bacillus cereus was identified as an infrequent abortigenic agent in cattle . Necrotizing placentitis with no or sporadic lesions in fetal tissues was seen . Bacillus cereus was isolated in pure culture from fetal tissue and/or placenta . The recent identification of a bovine abortion caused by B cereus, prompted a retrospective survey of the pathology files . Eight of 947 bovine abortions were attributed to B cereus . Bacillus cereus is often mistaken as a contaminant in bovine abortion because of the failure to identify lesions in fetal tissues compatible with bacterial invasion . A necrotizing toxin may be responsible for the placentitis, with expulsion of the fetus before bacterial colonization. J Appl Bacteriol, 1985 Nov, 59(5), 479 - 86 The release of dipicolinic acid during heating and its relation to the heat destruction of Bacillus stearothermophilus spores; Mallidis CG et al.; The rates of dipicolinic acid (Dpa) release and the rates of death were studied for spores of five strains of Bacillus stearothermophilus . It was observed that a highly significant relationship exists between the rate of Dpa release and rate of spore death for the four out of five strains tested and for all test temperatures . At 115 degrees C the rate of Dpa release was found to be faster than the rate of death, equal at 120 degrees C and slower at 125 degrees C . The role of Dpa in heat resistance was considered and a theory is proposed to explain the mechanism by which the heat resistance of bacterial spores is overcome. J Appl Bacteriol, 1985 Nov, 59(5), 407 - 11 Determination of the heat resistance of spores using a solid heating block system; Mallidis CG et al.; A simple and accurate technique for the determination of the heat resistance of spores is described . The technique combines a modified capillary tube method with a solid heating block . The come-up time of spore suspensions was found to be short and simple and accurate technique is suggested for the correction of the come-up times . Experimental results are presented for the destruction of spores of Bacillus stearothermophilus at 120 degrees which indicates the accuracy and reproducibility of the new method. J Dairy Sci, 1985 Nov, 68(11), 3031 - 6 Quantitative assay for antibiotics used commonly in treatment of bovine infections; Bishop JR et al.; Delvotest-P and Bacillus stearothermophilus Difco disc assay procedures were utilized, and assay sensitivity for detecting various antibiotics was tested . Bacillus stearothermophilus spores used in the disc assay were purchased from two laboratories and compared . The dyes methylene blue, 2,3,5-triphenyltetrazolium chloride, orange-O, and bromcresol purple were incorporated separately into agar used for the disc assay to determine if sharper, clearer zones could be produced . None of the dyes had an observable effect on zone size or clarity . Using the Difco disc assay, quantitation of penicillin, cephapirin, and oxytetracycline was accomplished with reliable precision by relating the size of the zone of inhibition to the log of the antibiotic concentration . The Delvotest-P and Difco disc assays were equal in sensitivity, as were spore suspensions used from the two laboratories. Equine Vet J, 1985 Nov, 17(6), 445 - 8 BCG treatment of periocular sarcoid; Lavach JD et al.; Twenty-six horses and five mules with periocular sarcoids were treated with intralesional injections of a purified bacillus of Calmette and Guerin (BCG) cell walls in oil suspension . All sarcoids were cured and the horses and mules remained free from recurrence of sarcoid during the two-year follow-up period. J Bacteriol, 1985 Nov, 164(2), 938 - 40 Nitrogen catabolite repression of the L-asparaginase of Bacillus licheniformis; Golden KJ et al.; We report the presence of a single L-asparagine aminohydrolase activity (EC 3.5.1.1) in extracts of Bacillus licheniformis A5 . The synthesis of the enzyme was apparently under nitrogen catabolite repression control. Acta Cytol, 1985 Nov-Dec, 29(6), 961 - 6 Fine needle aspiration cytology of granulomatous prostatitis induced by BCG immunotherapy of bladder cancer; Stilmant M et al.; Six patients treated with intravesical bacillus Calmette-Guerin (BCG) for superficial bladder cancer had granulomatous prostatitis demonstrated histologically by transperineal needle biopsy . Four of the six patients also underwent transrectal fine needle aspiration (FNA) for cytologic study . The diagnosis of granulomatous prostatitis was made cytologically in all four without knowledge of the histologic findings . Granulomatous prostatitis appears to be common following intravesical BCG treatment; these cases show that FNA cytology can be recommended as a method for diagnosing this complication. Can J Microbiol, 1985 Nov, 31(11), 994 - 9 Catalase, superoxide dismutase, and the production of O2-sensitive mutants of Bacillus coagulans; Vassilyadi M et al.; A number of facultatively anaerobic members of the genus Bacillus were screened for their catalase, diaminobenzidine peroxidase, and superoxide dismutase activities . A strain of Bacillus coagulans (7050) lacking peroxidatic activity and containing single catalatic and superoxide dismutase activities was selected . Responses of the superoxide dismutase activity and catalase level to the partial pressure of oxygen, and Fe and Mn levels, as well as to aerobic and fermentative metabolism, were determined . There appeared to be a relationship between high endogenous catalase levels and the high H2O2 evolution and KCN insensitivity of B . coagulans respiration . Bacillus coagulans 7050 was mutagenized with N-methyl-N'-nitro-N-nitrosoguanidine and screened for the expression of oxygen intolerance . All of the 38 stable oxygen sensitive mutants obtained had very low or completely absent catalatic activity and catalase protein . No mutant lacked superoxide dismutase, although five showed significantly lowered levels of the enzyme . Exogenous bovine liver catalase restored aerotolerance and reduced cell pleomorphism in the mutants. J Bacteriol, 1985 Nov, 164(2), 712 - 6 Export of alpha-amylase by Bacillus amyloliquefaciens requires proton motive force; Muren EM et al.; The secretion of protein directly into the extracellular medium by Bacillus amyloliquefaciens, a gram-positive bacterium, was shown to be dependent on proton motive force . When the electrochemical membrane potential gradient of protons was dissipated either by uncouplers or by valinomycin in combination with K+, a precursor form of alpha-amylase accumulated on the cellular membrane . The proton motive force could be dissipated without altering the intracellular level of ATP, indicating that the observed inhibition of export was not the result of decreased ATP concentration. J Immunol, 1985 Nov, 135(5), 3417 - 23 Relationship between membrane potential changes and superoxide-releasing capacity in resident and activated mouse peritoneal macrophages; Kitagawa S et al.; In an attempt to understand better the molecular basis for the enhanced respiratory burst of activated macrophages (M phi), we investigated the relationship between stimulus-induced changes in membrane potential and release of superoxide anion (O2-) in mouse peritoneal M phi . Resident M phi and M phi elicited by injection of lipopolysaccharide (LPS-M phi) or obtained from animals infected with bacille Calmette-Guerin (BCG-M phi) were used . LPS-M phi and BCG-M phi showed more pronounced changes in membrane potential (depolarization) and greater release of O2- on contact with phorbol myristate acetate (PMA) than did resident macrophages . The lag time between addition of stimulus and onset of release of O2- was reduced in activated compared with resident cells . Membrane potential changes began 60 to 90 sec before release of O2- could be detected in each cell type . The dose-response curves for triggering of membrane potential changes and O2- release by PMA were identical . The magnitude of membrane potential changes and of O2- release in LPS-M phi and BCG-M phi declined progressively during in vitro culture, and values on day 3 approached those in resident macrophages ("deactivation") . Extracellular glucose was required for effective stimulated change in membrane potential and O2- release . These findings indicate that membrane potential changes are closely associated with O2- -releasing capacity in macrophages, and that the systems that mediate membrane potential changes and production of O2- develop or decline concomitantly during activation or deactivation of the cells . Although the plasma membrane was highly depolarized by high extracellular K+ or by the sodium ionophore gramicidin, O2- release was not induced by these maneuvers, indicating that changes in membrane potential by themselves are not sufficient to trigger the respiratory burst in macrophages . Release of O2- was not impaired in buffers in which Na+ was completely replaced with equimolar concentrations of K+ or choline+; thus, induction or maintenance of the respiratory burst in M phi does not require an influx of Na+. Zhonghua Zhong Liu Za Zhi, 1985 Nov, 7(6), 432 - 4 {Effect of sodium selenite on the antitumor function of spleen lymphocytes in mice}; Wang LM; Sodium selenite in drinking water (1 ppm) was given to normal and hepatoma-bearing mice for 14 days . Spleen lymphocytes were incubated with hepatoma cells and 3H-TdR incorporation was measured . Sodium selenite increased the ability of spleen cells to inhibit 3H-TdR incorporation into hepatoma cells by more than 40% . In the meantime, in the spleen lymphocytes of normal and hepatoma-bearing mice, the activities of ATPase, acid phosphatase and alkaline phosphatase were definitely higher than that of the control (ATPase increased by 35.5% and 65.0%; acid phosphatase increased by 113.0% and 93.0%; alkaline phosphatase by 64.7% and 69.0%, respectively) . These results suggest that sodium selenite (1 ppm), like immunologic stimulator such as bacillus Calmette Guerin (BCG), be able to stimulate the immunologic function of organism against hepatoma. Appl Environ Microbiol, 1985 Nov, 50(5), 1196 - 9 Stability of the larvicidal activity of Bacillus thuringiensis subsp . israelensis: amino acid modification and denaturants; Pfannenstiel MA et al.; The Bacillus thuringiensis subsp . israelensis mosquito larvicidal toxin is not a sulfhydryl-activated toxin . The protein disulfide bonds were cleaved and blocked without loss of toxicity . In contrast, modification of the lysine side chains eliminated toxicity . Additionally, the toxin was resistant to high concentrations of salt (8 M NaBr), organic solvents (40% methanol), denaturants (4 M urea), and neutral detergents (10% Triton X-100) . However, it was inactivated by both positively and negatively charged detergents and by guanidine hydrochloride. J Biochem (Tokyo), 1985 Nov, 98(5), 1211 - 9 Regulation and properties of glutamine synthetase purified from Bacillus cereus; Matsuoka K et al.; The glutamine synthetase from Bacillus cereus IFO 3131 was purified to homogeneity . The enzyme is a dodecamer with a molecular weight of approximately 600,000, and its subunit molecular weight is 50,000 . Both Mg2+ and Mn2+ activated the enzyme as to the biosynthesis of L-glutamine, but, unlike in the case of the E . coli enzyme, the Mg2+-dependent activity was stimulated by the addition of Mn2+ . The highest activity was obtained when 20 mM Mg2+ and 0.5 mM Mn2+ were added to the assay mixture . For each set of optimal assay conditions, the apparent Km values for glutamate, ammonia and a divalent cation X ATP complex were 1.03, 0.34, and 0.40 mM (Mn2+: ATP = 1: 1); 14.0, 0.47, and 0.91 mM (Mg2+: ATP = 4: 1); and 9.09, 0.45, and 0.77 mM (Mg2+: Mn2+: ATP = 4: 0.2: 1), respectively . At each optimum pH, the Vmax values for these reactions were 6.1 (Mn2+-dependent), 7.4 (Mg2+-dependent), and 12.9 (Mg2+ plus Mn2+-dependent) mumoles per min per mg protein, respectively . Mg2+-dependent glutamine synthetase activity was inhibited by the addition of AMP or glutamine; however, this inhibitory effect was suppressed in the case of the Mg2+ plus Mn2+-dependent reaction . These results suggest that the activity of the B . cereus glutamine synthetase is regulated by both the intracellular concentration and the ratio of Mn2+/Mg2+ in vivo . Also in the present investigation, a potent glutamine synthetase inhibitor(s) was detected in crude extracts from B . cereus. J Biochem (Tokyo), 1985 Nov, 98(5), 1147 - 56 Complete nucleotide sequence of a gene coding for heat- and pH-stable alpha-amylase of Bacillus licheniformis: comparison of the amino acid sequences of three bacterial liquefying alpha-amylases deduced from the DNA sequences; Yuuki T et al.; The gene coding for the heat-stable and pH-stable alpha-amylase of Bacillus licheniformis 584 (ATCC 27811) was cloned in Escherichia coli and the nucleotide sequence of a DNA fragment of 1,948 base pairs containing the entire amylase gene was determined . As inferred from the DNA sequence, the B . licheniformis alpha-amylase had a signal peptide of 29 amino acid residues and the mature enzyme comprised 483 amino acid residues, giving a molecular weight of 55,200 . The amino acid sequence of B . licheniformis alpha-amylase showed 65.4% and 80.3% homology with those of heat-stable Bacillus stearothermophilus alpha-amylase and relatively heat-unstable Bacillus amyloliquefaciens alpha-amylase, respectively . Nevertheless, several regions of the alpha-amylases appeared to be clearly distinct from one another when their hydropathy profiles were compared. Biokhimiia, 1985 Nov, 50(11), 1859 - 65 {Limited proteolysis of human leukocyte interferon-alpha2 and localization of the antigenic determinant binding the monoclonal antibodies}; Kostrov SV et al.; Large peptide fragments of human leucocyte interferon-alpha 2 (INF-alpha 2) were obtained by limited proteolysis with trypsin, pepsin, thermolysine and Bacillus amyloliquefaciens intracellular serine proteinase . The ability of the fragments to bind murine monoclonal antibodies NK2 raised against INF-alpha 2 was studied by the immunoblotting technique . The region of sequence 110-149 is the most sensitive to proteolytic attack, being probably exposed on the surface of the INF-alpha 2 molecule . INF-alpha 2 fragments 1-139, 1-147, 1-149 are capable of binding antibodies, whereas fragments 1-109 and 1-112 do not bind antibodies NK2 . A comparison of the primary structure of human leucocyte and murine leucocyte INF families in the region of sequence 110-139 and an analysis of the ability of human INF differing in amino acid sequences to bind antibodies NK2 demonstrated that the antigenic determinant for antibodies NK2 is the sequence Glu114-Asp115-Ser116-Ile117 of the INF-alpha 2 molecule. Cell Immunol, 1985 Oct 15, 95(2), 443 - 9 Effect of methanol extraction residue tubercle bacillus fraction treatment in vivo and in vitro on the release and activity of suppressor factor inhibiting the efferent phase of contact sensitivity in mice; Zimber C et al.; BALB/c mice subjected to injection of dinitrobenzenesulfonate (DNBSO3) and skin painting with dinitrofluorobenzene (DNFB) produce splenic T-suppressor (Ts) cells that, when transferred to DNFB-sensitized recipients, suppress the efferent (eff) phase of contact sensitivity (CS) . Splenocyte populations containing such Ts-eff cells release a specific soluble suppressor factor (SSF) in vitro that similarly suppresses CS in sensitized recipient mice . Treatment with the methanol extraction residue (MER) tubercle bacillus fraction, a known immunomodulating agent, of animals exposed to DNBSO3 and DNFB ("DNBSO3-DNFB" donors) prevented release of SSF by their spleen cells at in vitro incubation . Incubation of DNBSO3-DNFB donor splenocytes with MER in vitro also abolished SSF release, and treatment with MER of test animals prior to DNFB sensitization prevented the suppressive action of subsequently administered SSF . The observations are discussed with regard to the potentiating capacities of MER on cellular immunity and states of resistance. Biochem Biophys Res Commun, 1985 Oct 15, 132(1), 19 - 27 Identification of the peptides of the crystals of Bacillus thuringiensis var israelensis involved in the mosquito larvicidal activity; Sriram R et al.; Tryptic digestion of the proteins from the purified crystals of B.thuringiensis var israelensis resulted in the decline of high molecular weight peptides without the loss of mosquito larvicidal activity, measured after immobilization of the digests with DEAE- Sephadex A 50 beads . Amongst the peptides generated (less than 44 kDa), a 21 kDa peptide was immunoreactive to the crystal antiserum . Analysis of the peptides released from spores of the toxic (Cry+) and non-toxic (Cry-) strains has revealed a pattern in which only the 26kDa peptide was missing in the Cry-strain . Sporulation and crystal formation were dissociated by the addition of the antibiotic netropsin, which could also inhibit the crystal assembly, without considerable decrease of the larvicidal activity and retention of the 26kDa peptide . These results implicate the 26kDa peptide in the larvicidal action. Biochemistry, 1985 Oct 8, 24(21), 5852 - 7 Reversible dissociation of dimeric tyrosyl-tRNA synthetase by mutagenesis at the subunit interface; Jones DH et al.; Dimeric tyrosyl-tRNA synthetase from Bacillus stearothermophilus exhibits half-of-the-sites reactivity and negative cooperativity in binding of tyrosine . Protein engineering has been applied to the enzyme to determine whether it can be reversibly dissociated into monomers and if the monomers are active . The target for mutation is the residue Phe-164 . The side chain of Phe-164 in one subunit interacts with its symmetry-related partner in the other . Mutation of Phe-164----Asp-164 gives a mutant {TyrTS(Asp-164)} that undergoes dissociation at high pH when the aspartate residues are ionized . The monomer is inactive and does not bind tyrosine . Dissociation is enhanced at low concentrations of enzyme by a mass action effect . Kinetic and binding measurements on TyrTS(Asp-164) with tyrosine and tyrosyl adenylate show that the monomer has very weak affinity for these ligands . Accordingly, dimerization is favored by high concentrations of tyrosine and ATP since the dimeric form has a high affinity for the ligands . The presence of tRNA does not encourage dimer formation, and so it must bind to the monomer . TyrTS(Asp-164) is fully active at pH 6 where dimerization is favored but has low activity at pH 7.8 where dissociation is favored . It should now prove possible to engineer heterodimers that may be used to investigate the subunit interactions further. Biochemistry, 1985 Oct 8, 24(21), 5858 - 61 Fine structure-activity analysis of mutations at position 51 of tyrosyl-tRNA synthetase; Fersht AR et al.; Residue Thr-51 at the active site of tyrosyl-tRNA synthetase (Bacillus stearothermophilus) has been replaced with all the smaller amino acids by protein engineering to investigate direct and indirect effects of mutation on substrate binding and catalysis . The gamma-hydroxyl group of Thr-51 was thought to be 0.5 A too far from the ribose ring oxygen of ATP to form a hydrogen bond . Consistent with this, it is found that mutation of Thr-51----Cys-51, which should place the gamma-thiol group within its correct distance for hydrogen bonding, increases the affinity of the enzyme for ATP . Other mutations (Ser-51, Ala-51, and Gly-51) show the contributions to binding of the other atoms in the side chain of Thr-51 . A family of enzymes has been produced, TyrTS(Thr-51) (wild type), TyrTS(Ala-51), TyrTS(Cys-51), and TyrTS(Pro-51), in which the value of kcat/KM for ATP in aminoacylation increases along the series . This is achieved by the value of KM decreasing significantly (2.5, 1.25, 0.29, and 0.019 mM, respectively) while there are smaller decreases in kcat (4.7, 4.0, 2.9, and 1.8 s-1, respectively) . These variations cause each one of the enzymes to be more active than the others at particular concentrations of ATP . For example, at concentrations of ATP greater than 5.9 mM, TyrTS(Thr-51) is the most active, while TyrTS(Ala-51), TyrTS(Cys-51), and TyrTS(Pro-51) are the most active at 5.9-2.2, 2.2-0.42, and less than 0.42 mM ATP, respectively . Interestingly, position 51 shows variation in tyrosyl-tRNA synthetases isolated from different organisms. Hinyokika Kiyo, 1985 Oct, 31(10), 1701 - 7 {Intravesical Bacillus Calmette-Guerin for treatment of bladder tumor}; Uchida T et al.; The patients with bladder tumor (11 tumor) were given intravesical Bacillus Calmette-Guerin (BCG) therapy . The concentration of BCG solution varied from 80 to 240 mg in 40 ml of normal saline . The intravesical instillation therapy with this solution was repeated 6 times at a week interval . Seven of the tumors showed excellent response and had completely disappeared upon endoscopic examination . All of the highly responded tumors were small papillary lesions of low grade and low stage of malignancy . Regarding side effects, irritable bladder occurred in 70%, elevation of body temperature of over 37 degrees C in 40% and gross hematuria in 30% of the patients . Among them, one patient was treated with INH (0.3 g/day) for 2 weeks with satisfactory remission of the symptoms . Though the number of cases and follow up period are not satisfactory enough, topical therapy with BCG can be said to be an effective therapeutic measure against early stage urotherial tumors. Mol Cell Biochem, 1985 Oct, 68(2), 131 - 7 Bacitracin increases size of parasporal crystals and spores in Bacillus thuringiensis; Garcia-Patrone M; The effect of bacitracin on Bacillus thuringiensis is described . When added after the end of the exponential phase, it produces a marked increase in the size of spores and parasporal crystals . The cultures to which the antibiotic was added are able to produce more crystal proteins than non-treated cultures . The protein composition of crystals from bacitracin-treated cultures is the same as that of crystals purified from control cultures. Urology, 1985 Oct, 26(4 Suppl), 18 - 26 Overview of treatment of superficial bladder cancer; Soloway MS; Seventy per cent to 80 per cent of patients with bladder cancer will have their initial tumor confined to the mucosa (Stage Ta or Tcis, carcinoma in situ) or lamina propria (Stage T1), and at least 50 per cent of them will have a true recurrence or new recurrence despite resection of their initial tumor . If the tumor is of low grade and confined to the mucosa, intravesical chemotherapy might be considered either as treatment if the neoplasm is too extensive to resect completely or if multiple prior endoscopic resections have failed, or, alternatively, as prophylaxis against a subsequent tumor after complete endoscopic removal of the existing neoplasm . Thiotepa, until recently the most commonly used agent, completely eradicates all obvious tumor in approximately 30 per cent of patients, and it is effective in preventing subsequent tumor . Myelosuppression occurs in up to 20 per cent of patients receiving this agent, so careful monitoring of the white blood cell and platelet counts is mandatory . Mitomycin has low risk of myelosuppression and is effective in both the treatment and prophylaxis of superficial bladder cancer; the complete response rate is the same whether patients had an initial low-grade papillary or high-grade tumor (carcinoma in situ) . Doxorubicin hydrochloride and bacillus Calmette-Guerin (BCG) are other agents that have been used for intravesical therapy . Chemical cystitis, the primary side effect of doxorubicin, has caused discontinuation of treatment in 15 per cent to 20 per cent of patients . The efficacy of doxorubicin varies among those reporting its use . BCG has been shown to be effective in both treatment and prophylaxis, with response rates similar to those reported for mitomycin; however, a comparative trial has not been performed . There is a need to standardize the potency of each BCG vial and to determine fully the necessity of concomitant intradermal administration. Exp Parasitol, 1985 Oct, 60(2), 239 - 44 Nematoda: susceptibility of the egg to Bacillus thuringiensis toxins; Bottjer KP et al.; Crystalline toxins from Bacillus thuringiensis israelensis and B.t . kurstaki were lethal in vitro to eggs of the nematode Trichostrongylus colubriformis . The LD50 values for the two toxins were 0.38 ng and 37.5 micrograms total protein/ml, respectively . After 1 week at ambient temperature, the LD50 of B.t . kurstaki decreased to less than 4 micrograms/ml . Toxin from B.t . israelensis had no effect within 48 hr on survival of adult nematodes or on their feeding in vitro . Third-stage larvae of T . colubriformis were also unaffected by B.t . israelensis toxin . Exposure of third-stage larvae of Nippostrongylus brasiliensis to 1.1 micrograms total protein/ml of B.t . israelensis for 4 hr had no effect on their infectivity in mice, based on recovery of helminths at 7 days after infection . Similar exposure of 5-day-old N . brasiliensis and subsequent transfer into the intestine of mice gave recoveries that were similar to the untreated control . Thirty strains of B . thuringiensis caused mortality in nematode eggs, but over a 77,000-fold range of activity was found, based on the LD50 values . Toxin from B.t . israelensis was lethal to eggs of six zooparasitic and one free-living species of nematode, but the LD50 values varied 28-fold . Addition of B.t . israelensis to feces that contained eggs of T . colubriformis reduced subsequent recovery of larvae, with an LD50 of 260 ng/g of feces. J Ultrastruct Res, 1985 Oct-Nov, 93(1-2), 27 - 32 Light-triggered organization of the stigma in dark-grown nondividing cells of Euglena gracilis; Osafune T et al.; Dark-grown cells of Euglena gracilis Klebs var . bacillaris Cori contain amorphous stigma material . When these cells are placed on resting medium for 3 days in darkness, the cells cease division; the organization of a normal stigma from the amorphous material requires continuous illumination for 72-96 hr . We have now found that if dark-grown cells are placed on resting medium for 8 days, a 40-min light pulse is sufficient to cause normal organization of the stigma in a subsequent 72-hr dark period . Thus stigma development is light-dependent at 3 days of resting but becomes light-triggered at 8 days . Other examples of light-triggered phenomena in Euglena are discussed and a model based on turnover of protein molecules repressing development that are ordinarily removed by exposure to light is presented; it is suggested that as the cells become more starved their ability to replace repressor molecules removed by light becomes limited and the system thereby becomes light-triggered rather than light-dependent. Mikrobiyol Bul, 1985 Oct, 19(4), 235 - 7 {Food poisoning caused by Bacillus cereus}; Doganay M et al.; An outbreak of food poisoning was observed in Sivas on March 27, 1985 involving 25 patients who work at the same place . Clinical picture was characterized with nausea, abdominal pain and watery diarrhea . B . cereus was isolated from the stool of 4 patients . No other enteropathogens were found. Biochem J, 1985 Oct 1, 231(1), 83 - 8 Single-turnover and steady-state kinetics of hydrolysis of cephalosporins by beta-lactamase I from Bacillus cereus; Bicknell R et al.; The kinetics of the hydrolysis of two cephalosporins by beta-lactamase I from Bacillus cereus 569/H/9 has been studied by single-turnover and steady-state methods . Single-turnover kinetics could be measured over the time scale of minutes when cephalosporin C was the substrate . The other substrate, 7-(2',4'-dinitrophenylamino)deacetoxycephalosporanic acid, was hydrolysed even more slowly, and has potential for use in crystallographic studies of beta-lactamases . Comparison of single-turnover and steady-state kinetics showed that, for both substrates, opening the beta-lactam ring (i.e . acylation of the enzyme) was the rate-determining step . Thus the non-covalent enzyme-substrate complex is expected to be the intermediate observed crystallographically. J Bacteriol, 1985 Oct, 164(1), 302 - 9 Trypsinlike enzymes from dormant and germinated spores of Bacillus cereus T and their possible involvement in germination; Boschwitz H et al.; Trypsin-like enzymes were studied in dormant, activated, and germinated spores of Bacillus cereus T . Dormant spores contained two heat-labile enzyme activities . One was extractable with 2 M KCl and hydrolyzed azo-albumin . The second, a trypsinlike activity, was not extractable with 2 M KCl and hydrolyzed benzoyl-L-arginine-p-nitroanilide . Because of their heat instability, these two enzyme activities are probably not involved in the germination of heat-activated spores . Upon germination of heat-treated spores, a trypsinlike protease which was not detected in intact dormant spores was activated or exposed . This enzyme, when measured in intact germinated spores, hydrolyzed benzoyl-DL-arginine-p-nitroanilide but not azo-albumin and was inhibited in situ by sulfhydryl-blocking reagents such as p-chloromercuribenzoic acid and Hg2+ . There was a correlation between the inhibition of germination and enzymatic activity by sulfhydryl-blocking reagents . The enzyme was also inhibited by leupeptin, tosyl-L-lysine chromoethyl ketone, and tosyl-L-arginine methyl ester . Good correlation existed between the inhibition of germination and enzymatic activity by these agents . Electron micrographs showed that in the presence of trypsin inhibitors, the spores did not lose their cortex . The protein extracts of the inhibited spores formed a somewhat different electrophoretic pattern in sodium dodecyl sulfate-polyacrylamide gel electrophoresis than the protein extracts of dormant or germinated spores. J Bacteriol, 1985 Oct, 164(1), 223 - 9 Cloning and sequencing of the metallothioprotein beta-lactamase II gene of Bacillus cereus 569/H in Escherichia coli; Hussain M et al.; The structural gene for beta-lactamase II (EC 3.5.2.6), a metallothioenzyme, from Bacillus cereus 569/H (constitutive for high production of the enzyme) was cloned in Escherichia coli, and the nucleotide sequence was determined . This is the first class B beta-lactamase whose primary structure has been reported . The amino acid sequence of the exoenzyme form, deduced from the DNA, indicates that beta-lactamase II, like other secreted proteins, is synthesized as a precursor with a 30-amino acid N-terminal signal peptide . The pre-beta-lactamase II (Mr, 28,060) is processed in E . coli and in B . cereus to a single mature protein (Mr, 24,932) which is totally secreted by B . cereus but in E . coli remains intracellular, probably in the periplasm . The expression of the gene in E . coli RR1 on the multicopy plasmid pRWHO12 was comparable to that in B . cereus, where it is presumably present as a single copy . The three histidine residues that are involved (along with the sole cysteine of the mature protein) in Zn(II) binding and hence in enzymatic activity against beta-lactams were identified . These findings will help to define the secondary structure, mechanism of action, and evolutionary lineage of B . cereus beta-lactamase II and other class B beta-lactamases. Appl Environ Microbiol, 1985 Oct, 50(4), 737 - 42 Protease activation of the entomocidal protoxin of Bacillus thuringiensis subsp . kurstaki; Andrews RE Jr et al.; Two isolates of Bacillus thuringiensis subsp . kurstaki were examined which produced different levels of intracellular proteases . Although the crystals from both strains had comparable toxicity, one of the strains, LB1, had a strong polypeptide band at 68,000 molecular weight in the protein from the crystal; in the other, HD251, no such band was evident . When the intracellular proteases in both strains were measured, strain HD251 produced less than 10% of the proteolytic activity found in LB1 . These proteases were primarily neutral metalloproteases, although low levels of other proteases were detected . In LB1, the synthesis of protease increased as the cells began to sporulate; however, in HD251, protease activity appeared much later in the sporulation cycle . The protease activity in strain LB1 was very high when the cells were making crystal toxin, whereas in HD251 reduced proteolytic activity was present during crystal toxin synthesis . The insecticidal toxin (molecular weight, 68,000) from both strains could be prepared by cleaving the protoxin (molecular weight, 135,000) with trypsin, followed by ion-exchange chromatography . The procedure described gave quantitative recovery of toxic activity, and approximately half of the total protein was recovered . Calculations show that these results correspond to stoichiometric conversion of protoxin to insecticidal toxin . The toxicities of whole crystals, soluble crystal protein, and purified toxin from both strains were comparable. J Clin Invest, 1985 Oct, 76(4), 1514 - 21 Bacillus Calmette-Guérin-stimulated neutrophils release chemotaxins for monocytes in rabbit pleural spaces and in vitro; Antony VB et al.; Neutrophils are often seen first at sites of granulomatous inflammation but their contribution to monocyte recruitment and granuloma formation is unknown . We tested the hypothesis that neutrophils release chemotaxins which attract monocytes . We found that rapid accumulations of fluid and influxes of neutrophils followed by monocytes occurred in bacillus Calmette--Guerin (BCG)-sensitized rabbits given BCG intrapleurally but did not occur in nitrogen mustard-treated (neutropenic) BCG-sensitized rabbits given BCG intrapleurally--unless the rabbits were also given intrapleural injections of neutrophils . We also found monocyte chemotaxins in pleural spaces of control and neutrophil-reconstituted neutropenic but not in neutropenic rabbits given BCG intrapleurally . Moreover, pleural fluid monocyte chemotaxins had molecular weights (12,000-15,000 and 1,000) that were similar to molecular weights of monocyte chemotaxins present in supernatants from mixtures of neutrophils and BCG in vitro . In addition, intrapleural injection of neutrophils and BCG or supernatants from in vitro mixtures of neutrophils and BCG (but not neutrophils or BCG alone) increased the numbers of monocytes and 3H cell pellet activity in pleural fluids from untreated neutropenic rabbits or neutropenic rabbits previously injected intravenously with 3{H}methyl thymidine-labeled monocytes . Furthermore, fewer BCG were recovered from pleural fluids of BCG-sensitized control compared to neutropenic rabbits given BCG, and at autopsy 10 d after instillation of BCG, control but not neutropenic rabbits had well-defined granulomas without adhesions on their pleural surfaces . Our results suggest that BCG stimulates neutrophils to release chemotaxins that recruit monocytes, and that these responses might contribute to granuloma formation in tuberculous pleurisy. Immunology, 1985 Oct, 56(2), 377 - 9 Susceptibility and HLA-B27 in post-dysenteric arthropathies; van Bohemen CG et al.; A recent outbreak of bacillary dysentery in The Netherlands revealed that, despite the close association of HLA-B27 with post-dysenteric or reactive arthritis (ReA), not even in one family did all HLA-B27 positive patients infected by an arthritogenic bacterium, develop ReA . This dissociation shows that additional factors beside B27 may determine susceptibility to ReA. J Bacteriol, 1985 Oct, 164(1), 107 - 13 Immunoelectron microscopic double labeling of alkaline phosphatase and penicillinase with colloidal gold in frozen thin sections of Bacillus licheniformis 749/C; Guan T et al.; The subcellular distribution of alkaline phosphatase and penicillinase was determined by double labeling frozen thin sections of Bacillus licheniformis 749/C with colloidal gold-immunoglobulin G (IgG) . Antipenicillinase and anti-alkaline phosphatase antibodies were used to prepare complexes with 5- and 15-nm colloidal gold particles, respectively . The character of the labeling of membrane-bound alkaline phosphatase and penicillinase was different: the immunolabels for alkaline phosphatase (15-nm particles) were bound to a few sites at the inner surface of the plasma membrane, and the gold particles formed clusters of various sizes at the binding sites; the immunolabels for penicillinase (5-nm particles), on the other hand, were bound to the plasma membrane in a dispersed and random fashion . In the cytoplasm, immunolabels for both proteins were distributed randomly, and the character of their binding was similar . The labeling was specific: pretreating the frozen thin sections with different concentrations of anti-alkaline phosphatase or penicillinase blocked the binding of the immunolabel prepared with the same antibody . Binding could be fully blocked by pretreatment with 800 micrograms of either antibody per ml. Appl Environ Microbiol, 1985 Oct, 50(4), 984 - 8 Micro-lipid-droplet encapsulation of Bacillus thuringiensis subsp . israelensis delta-endotoxin for control of mosquito larvae; Cheung PY et al.; The crystal delta-endotoxin of Bacillus thuringiensis subsp . israelensis is less toxic to larvae of Anopheles freeborni than to larvae of Aedes aegypti . However, when solubilized crystal was used, larvae from both species showed similar sensitivities . This effect presumably was due to the differences in feeding behavior between the two mosquito larvae when crystal preparations are used . A procedure is described whereby both crystal and solubilized B . thuringiensis subsp . israelensis toxin were emulsified with Freund incomplete adjuvant, with retention of toxicity . The use of Freund incomplete adjuvant also allowed one to assay the solubilized toxin at a low nanogram level . Furthermore, coating the toxin with lipophilic material altered the buoyancy of the toxin and reversed the sensitivities of the two mosquito larvae toward the B . thuringiensis subsp . israelensis toxin . This difference in buoyancy was determined by using an enzyme-linked immunosorbent assay that was specific for the toxic peptides . These data indicate that economically feasible buoyant formulations for the B . thuringiensis subsp . israelensis crystal can be developed. Immunology, 1985 Oct, 56(2), 235 - 43 Effect of Bacillus Calmette-Guérin on the in vitro generation of cytotoxic T lymphocytes . II . Role of interleukin-1-like factors and of soluble suppressor factors; Mellow L et al.; The injection of BCG vaccine in C57BL/6J mice results in the suppression of the generation of cytotoxic T lymphocytes (CTL) in mixed lymphocyte cultures (MLC) and of mitogenic reactions to concanavalin A (Con A) . Suppression is mediated by macrophage-like suppressor cells . Since previous work had indicated that suppression involved the inhibition of the production of interleukin-2 (IL-2), the effects of BCG on interleukin-1 (IL-1), a monokine required for IL-2 production, were investigated . It was found that the release of IL-1-like activity in spleen cell cultures stimulated with LPS or Con A was increased by previous BCG treatment of the cell donors . In MLC, the release of IL-1-like activity was also increased by BCG . However, the detection of IL-1-like activity in MLC supernatants was prevented by the presence of a suppressor factor . In this case, the IL-1-like activity could be separated with gel filtration from the suppressor factor which had higher molecular weight . The production of IL-1-like activity by CBA/J spleen cells, which are not suppressed by BCG, was not significantly different from that of C57BL/6J cells, which are markedly suppressed . Moreover, the addition of IL-1 to the BCG-suppressed cultures not only did not restore normal reactivity, but actually further suppressed CTL formation . It was concluded that BCG-induced suppression cannot be attributed to decreased IL-1 activity . The suppressor factor discovered during these investigations may have a role in this type of suppression. J Infect Dis, 1985 Oct, 152(4), 811 - 6 Efficacy of ciprofloxacin in experimental arthritis caused by Escherichia coli--in vitro-in vivo correlations; Bayer AS et al.; Ciprofloxacin, a new carboxyquinolone, has potent in vitro bactericidal activity against the major aerobic, gram-negative bacillary pathogens that cause human pyoarthroses . We investigated the in vivo efficacy of ciprofloxacin in a rabbit model of septic arthritis due to Escherichia coli . Animals received either ciprofloxacin (80 mg/kg per day) or gentamicin (5 mg/kg per day) . Ciprofloxacin was rapidly bactericidal in vivo and was significantly more effective in reducing the numbers of E . coli in synovial tissue than was gentamicin at days 10 and 17 of therapy (P less than .0005 and P less than .05, respectively) . Similarly, ciprofloxacin was significantly more active than was gentamicin in reducing the numbers of E . coli in joint fluid on day 10 of therapy (P less than .0005); however, by day 17 of therapy, the numbers of E . coli in joint fluid were not significantly different in the two therapy groups . Neither regimen was effective in preventing the development of postinfectious inflammatory synovitis . There was no in vivo development of resistance to either antibiotic during therapy . Ciprofloxacin therapy was associated with significantly higher bactericidal titers in serum and joint fluid than were observed with gentamicin therapy (P less than .0005) . Ciprofloxacin warrants further in vivo evaluation in invasive E . coli infections. J Biol Chem, 1985 Sep 25, 260(21), 11393 - 5 Reconstitution of a bacterial Na+/H+ antiporter; Seto-Young D et al.; Membrane proteins from alkalophilic Bacillus firmus RAB were extracted with octylglucoside, reconstituted into liposomes made from alkalophile lipids . The proteoliposomes were loaded with 22Na+ . Imposition of a valinomycin-mediated potassium diffusion potential, positive out, resulted in very rapid efflux of radioactive Na+ against its electrochemical gradient . That the Na+ efflux was mediated by the electrogenic Na+/H+ antiporter is indicated by the following characteristics that had been established for the porter in previous studies: dependence upon an electrical potential; pH sensitivity, with activity dependent upon an alkaline pH; inhibition by Li+; and an apparent concentration dependence upon Na+ that correlated well with measurements in cells and membrane vesicles. FEBS Lett, 1985 Sep 23, 189(2), 207 - 11 The amino acid sequence of the zinc-requiring beta-lactamase II from the bacterium Bacillus cereus 569; Ambler RP et al.; The amino acid sequence of the zinc-requiring beta-lactamase II from Bacillus cereus strain 569 has been determined . It consists of a single polypeptide chain of 227 residues . It is the only example so far fully characterized of a class B beta-lactamase, and is structurally and mechanistically distinct from both the widely distributed class A beta-lactamases (such as the Escherichia coli RTEM enzyme) and from the chromosomally encoded class C enzymes from Gram-negative bacteria. Biochim Biophys Acta, 1985 Sep 20, 831(1), 133 - 41 Charged detergents enhance the activity of phospholipase C (Bacillus cereus) towards micellar short-chain phosphatidylcholine; el-Sayed MY et al.; Phospholipase C (phosphatidylcholine cholinephosphohydrolase, EC 3.1.4.3) (Bacillus cereus) activity toward diheptanoylphosphatidylcholine is increased 50-100% by low concentrations of both positively and negatively charged detergents . Zwitterionic and nonionic detergents have no such activating effect . This charged detergent activation requires an interface, since comparable detergent concentrations have no effect on the hydrolysis rate of monomeric dihexanoylphosphatidylcholine . From NMR and diacylglycerol solubility studies it is suggested that activation results from detergent interacting with diacylglycerol to accelerate product release from the enzyme. Biochem J, 1985 Sep 15, 230(3), 829 - 32 The association of penicillin-binding proteins with cell elongation and septum formation in Bacillus megaterium; Todd JA et al.; Analysis of the penicillin-binding proteins in the membranes from germinated spores of Bacillus megaterium and a filamentous mutant indicated that (a) no specific synthesis of any penicillin-binding protein occurred before or during two relatively synchronous cell divisions, (b) penicillin-binding proteins 1, 3a and 3b may be involved in cell elongation and (c) filamentation of the mutant may be due to a decrease in the concentration of penicillin-binding protein 1. Biochemistry, 1985 Sep 10, 24(19), 5106 - 9 Probing histidine-substrate interactions in tyrosyl-tRNA synthetase using asparagine and glutamine replacements; Lowe DM et al.; We have analyzed the interactions of a histidine residue with a substrate using site-directed mutagenesis . Previous studies on tyrosyl-tRNA synthetase from Bacillus stearothermophilus have shown that a histidine residue (His-48) makes an interaction with ATP, which is improved on mutating Thr-51----Pro-51 . We find on replacing His-48 in wild-type enzyme with either asparagine or glutamine that Asn-48 is equally as good as His-48 but His-48----Gln-48 leads to a far lower activity . The side chain of an asparagine residue may be superimposed on that of a histidine so that the amide-NH2 group of asparagine occupies the same position as the pi-N of histidine, whereas the equivalent -NH2 group of glutamine may be superimposed upon the tau-N . This suggests that it is the pi-N of histidine that hydrogen bonds with ATP and that there is no significant electrostatic interaction between the histidine and ATP . Incorporating the Pro-51 mutation into each of the Asn-48 and Gln-48 mutants gives an improvement in the affinity of the enzyme for ATP, but this improvement is less than that seen with the wild-type enzyme. Mikrobiologiia, 1985 Sep-Oct, 54(5), 770 - 3 {Possible means for increasing exotoxin synthesis by a Bacillus thuringiensis culture--the producer of bitoxibacillin}; Abrosimova LI et al.; Various procedures were studied for obtaining Bacillus thuringiensis strains of serotype I which synthesized exotoxins . Mutant clones with elevated exotoxin synthesis could be selected by treating the cells with N-methyl-N-nitro-N-nitrosoguanidine . The frequency of (+) variant selection was from 17 to 12 X 10(-2) . The clones of S and R types differed in the insecticide activity of the exotoxin . Its yield could be increased by optimizing the composition of growth media . The strain specificity of B . thuringiensis producing the exotoxin was assayed in terms of carbon and nitrogen requirements. J Assoc Off Anal Chem, 1985 Sep-Oct, 68(5), 1018 - 20 Antibiotic identification by high voltage electrophoresis bioautography; Lott AF et al.; The high voltage electrophoresis bioautography method is applicable to meat, milk, and animal feeds . Meat is freeze-dried, powdered, and extracted with acetonitrile-water (9 + 1), and the extract is concentrated by evaporation at room temperature . Milk is examined directly or following acetonitrile-water extraction . Feed is extracted with acetonitrile-water . Samples or extracts are applied to preliminary assay plates of antibiotic medium No . 1 at pH 6 and 8, seeded with Micrococcus luteus (ATCC 9341), M . luteus DHSR (ATCC 9341A), Bacillus cereus (ATCC 11778), or B . cereus K250 TR (NCIB 11183), and nutrient agar at pH 7 seeded with B . subtilis BGA . Inactivation of penicillinase indicates beta-lactam antibiotics . Addition of trimethoprim increases sensitivity to sulfonamides . After 18-24 h incubation at 30 degrees C, plates yielding clear inhibition zones guide selection of conditions for subsequent electrophoresis bioautography . Extracts are applied (5-100 microL) to 10 mm diameter wells on electrophoresis plates 60 cm long and 40 cm wide, with a gel depth of 1.6 mm . The support medium is 1% agar and 1% agarose in Tris/succinic acid buffers pH 6 and pH 8 . A potential of 1500 V is applied for 1.5 h at 15 degrees C . Following electrophoresis, the migrated antibiotics are visualized by over-layering with antibiotic medium No . 1, pH 6 or 8, seeded with M . luteus or B . cereus spore suspension; plates are incubated for 18-24 h at 30 degrees C . Identification is based on results of preliminary screening together with electrophoretic migration distances and inhibition zone appearances compared with standards. J Urol, 1985 Sep, 134(3), 531 - 2 Durable response of a carcinoma in situ of the renal pelvis to topical bacillus Calmette-Guerin; Herr HW; The first long-term remission of high grade, flat carcinoma in situ of the renal pelvis treated with topical bacillus Calmette-Guerin is presented. Clin Immunol Immunopathol, 1985 Sep, 36(3), 275 - 88 Binding, endocytosis, and degradation of model immune complexes by murine macrophages at various levels of activation; Finbloom DS; To determine if stimulation of macrophages enhances their capacity to recognize and process immune complexes, the binding, endocytosis, and degradation of soluble model immune complexes in murine macrophages have been measured . These complexes are composed of covalently crosslinked dimers and heavy oligomers of murine IgG antidinitrophenyl antibodies . Resident (unstimulated), inflammatory (thioglycolate-elicited), and activated (bacillus Calmette-Guerin-elicited) macrophages were studied . Cells elicited with either agent bound five times more dimers or heavy oligomers than did resident cells . Stimulated macrophages internalized and then degraded 50-60% of those heavy oligomers initially bound to the surface of the cell . Resident macrophages, however, were about 70% as efficient as stimulated cells at degrading intracellular complexes . Although dimers avidly bound to all three types of macrophages, they were internalized poorly and were degraded minimally . Intracellular complexes were degraded within lysosomes as determined by centrifugation of cell homogenates on Percoll gradients . Following endocytosis of heavy oligomers, Fc receptors on each type of macrophage were down-modulated to 30% of control levels. Mycopathologia, 1985 Sep, 91(3), 159 - 63 Prophage induction and filamentation in Bacillus thuringiensis caused by the genotoxic mycotoxin aflatoxin B1; Auffray Y et al.; Cultures of the lysogenic strain of Bacillus thuringiensis var . tolworthi were made in the presence of various drugs . The determination of bacterial size and plaque forming units (by using an indicator strain of B . thuringiensis var . galleriae) as well as colony forming units were then performed . Treatment of lysogenic cells by aflatoxin B1: provokes the formation of elongated cells (filamentation); induces a pathway that leads to the induction of prophage . Results of the present study indicated that filament formation and bacteriophage induction are two commonplace effects that occur in virtually every member of this cellular population exposed to low doses of certain drugs such as aflatoxin B1 (10 micrograms/ml); all of which have in common the ability to produce damaging changes in DNA . The following findings support the hypothesis that error-prone repair mechanisms seem to be present in B . thuringiensis as in Escherichia coli. J Exp Med, 1985 Sep 1, 162(3), 917 - 29 An analysis of in vitro T cell responsiveness in lepromatous leprosy; Kaplan G et al.; In lepromatous leprosy, there is extensive replication of Mycobacterium leprae (M . leprae) within dermal macrophages . This lack of microbial resistance has been attributed to a defective cell-mediated immune response to M . leprae antigens . We have examined the in vitro response of T cells to M . leprae to determine if hyporesponsiveness could be reversed . The study included 40 unselected patients from New York and from Colombia, most with the severe lepromatous form of the disease . We first noted that lepromatous leprosy patients were of two types: those unable to respond, as assessed by T cell proliferation and immune (gamma) interferon (IFN-gamma) release, and a second group, exhibiting low but detectable responses relative to tuberculoid controls . When the effect of exogenous recombinant interleukin-2 (IL-2) on the response to M . leprae antigens was compared in the two groups, many of the low responders, but not the nonresponders, showed enhanced proliferation and IFN-gamma release . To evaluate a possible suppressive effect of monocytes, these cells were eliminated with a cell-specific monoclonal antibody and complement . Depletion of monocytes often expanded preexisting weak responses but did not reverse the anergy of the M . leprae nonresponders . The enhancement was not M . leprae-specific, since it was also observed when bacillus Calmette-Guerin was the antigenic stimulus for proliferation and IFN-gamma production . Removal of the suppressor T cell subset, with OKT8 antibody and complement, also did not restore responses in nonresponder patients . We conclude that a sizable number of lepromatous leprosy patients exhibit a low degree of responsiveness to M . leprae and that the responses can be enhanced in vitro with IL-2 or with monocyte depletion . Nonresponsiveness, however, cannot be reversed . Since currently available assays measure the function of previously sensitized T cells, suppressor mechanisms may yet contribute to defective cell-mediated immunity by impairing the initial sensitization to M . leprae antigens. Ann Inst Pasteur Immunol, 1985 Sep-Oct, 136D(2), 151 - 62 Vaccines against mycobacteria and other intracellular multiplying bacteria; Lagrange PH et al.; As the prototype of a vaccine against mycobacterial infection, the BCG (bacille bilie Calmette et Guerin) has been used against tuberculosis for more than 60 years . It is the only live attenuated vaccine used on humans in more than 182 countries or territories in the world, and very few changes have been made in its fabrication and distribution, except for the production of lyophilized seed-lots . However, its history is marked with controversies concerning its innocuity and efficacy . While BCG safety is no longer a matter of debate, the question of its effectiveness is still pertinent, and results in several controlled trials have shown great variability (from 0 to 80%) . The studies of different variables involved in such results have shown statistical bias, and numerous factors are involved in the highly complex interrelationships between the host, the pathogen and environmental factors . World-wide research is now being conducted under the auspices of the World Health Organisation, in order to gain further knowledge of the immunology of tuberculosis and leprosy . Such results are aimed at understanding variations in BCG efficacy and producing strategies for developing new vaccines and alternative methods for prophylaxis and diagnosis . Concerning human infections due to other facultative intracellular multiplying bacteria, there are relatively few vaccines which are able to give long-lasting and efficient protection . Some controversy remains as to the live attenuated mutant GalE S . typhi Ty21a, and there is hope for the new insoluble phenol extract from Brucella abortis, strain B19 . Further research is necessary on the others, for instance, Listeria monocytogenes, Chlamydia trachomatis and Legionella sp. EMBO J, 1985 Sep, 4(9), 2389 - 95 Characteristic views of E . coli and B . stearothermophilus 30S ribosomal subunits in the electron microscope; van Heel M et al.; Large sets of electron microscopic images of the 30S ribosomal subunits of Bacillus stearothermophilus (914 molecules) and Escherichia coli (422 molecules) were analysed with image processing techniques . Using computer alignment and a new multivariate statistical classification scheme, three predominant views of the subunit were found for both species . These views, which together account for approximately 90% of the population of images, were determined to a reproducible resolution of up to 1.7 nm, thus elucidating many new structural details . The angular spread of the molecular orientations around the three main stable positions is remarkably small (less than 8 degrees) . Some of the current models for the small ribosomal subunit are incompatible with our new results. Antibiot Med Biotekhnol, 1985 Sep, 30(9), 662 - 5 {Use of beta-lactamase from Bacillus licheniformis 749/C for solid-phase immunoenzyme analysis}; Levi MI et al.; beta-Lactamase from Bacillus licheniformis 749/c was used as a marker for ELISA . Conjugates of the beta-lactamase with the capsule antigen of the plague causative agent and with the monoclonal antibodies to it were prepared by glutaric aldehyde "linking" . The conjugates preserved high immunospecific and beta-lactamase activity . High sensitivity of the modification of ELISA allowed detecting up to 8 ng/ml of the capsule antigen of the plague causative agent . The enzymatic activity of the beta-lactamase was determined microiodometrically which provided visual recording of the results of ELISA. Int J Lepr Other Mycobact Dis, 1985 Sep, 53(3), 447 - 54 Isolation of characteristic glycolipids possibly included in spherical droplets around M . leprae; Fukunishi Y et al.; The main purpose of this work was to isolate the components in acetone soluble lipids of lepromas of the nine-banded armadillo by high performance liquid chromatography (HPLC), and then to examine the mass spectrometric characteristics of the two peaks (molecular weights 2000 and 1600) found by HPLC . The armadillo had been inoculated with Mycobacterium leprae isolated from a mangabey monkey with naturally acquired leprosy . According to the results of HPLC, gas liquid chromatographic and mass spectral analyses, the GPC peak I lipid at 2000 D was identified as phenolic glycolipid and the GPC peak II lipid at 1600 D, as phthiocerol dimycocerosate . It was thought that the GPC peak I lipid and the GPC peak II lipid were included in the spherical droplets (peribacillary substance) around M . leprae . It was concluded that the microorganisms causing leprosy-like changes in the mangabey monkey were either M . leprae or a very closely related bacillus. J Am Mosq Control Assoc, 1985 Sep, 1(3), 316 - 9 Evaluation of Beecomist-applied Bacillus thuringiensis (H-14) against Anopheles quadrimaculatus larvae in rice fields; Sandoski CA et al.; The Beecomist spray head was evaluated for aerial application of Bacillus thuringiensis var . israelensis (serotype H-14; Bti) at various ultra low volume (ULV) rates against natural populations of Anopheles quadrimaculatus larvae in rice fields . Deposits on Kromekote cards indicated that 0.54 liter/ha of neat Bti penetrated the dense canopy of the rice field . Mean number of droplets 65 cm below canopy level was 4.9 +/- 5.0/100 cm2 . At 1 day posttreatment, applications of 0.54, 0.27, 0.11, 0.07 and 0.04 liter of Bti/ha resulted in reductions of 97.9, 94.4, 93.0, 71.1 and 21.8%, respectively, of An . quadrimaculatus larvae/dip . Calculated lethal field dosages (LFD50 and LFD90) were 0.05 and 0.16 liter/ha, respectively. J Am Mosq Control Assoc, 1985 Sep, 1(3), 369 - 71 Five new mosquito larvicidal strains of Bacillus sphaericus from non-mosquito origins; Lysenko O et al.; Five new strains of Bacillus sphaericus having larvicidal activity similar to that of strains 1593 and 2362 are described . These strains were isolated from caterpillars or grasshoppers, but have no insecticidal activity toward these insects. J Am Mosq Control Assoc, 1985 Sep, 1(3), 310 - 5 Efficacy and field evaluation of Bacillus thuringiensis (H-14) and B . sphaericus against floodwater mosquitoes in California; Mulla MS et al.; The microbial control agents Bacillus thuringiensis (H-14) and B . sphaericus were evaluated in laboratory and field against Psorophora columbiae . Bacillus sphaericus strain 2362 was also tested in the field against Aedes melanimon . Psorophora columbiae was slightly more susceptible than Culex quinquefasciatus to active strains of B . sphaericus . The LC90 for active strains ranged from 0.013 to 0.069 mg/liter . In field trials, aqueous suspensions of primary powder of B . sphaericus 2362 and 1593 yielded 98-99% reduction in larvae at the rates of 0.1 to 0.25 lb/acre of the primary powder . Granular formulations of Bt (H-14) were evaluated against Ps . columbiae, yielding 96-99% control of larvae at rates ranging from 1 to 10 lb/acre of the granules, depending on the potency and type of the formulations . Aedes melanimon was slightly less susceptible than Ps . columbiae to B . sphaericus 2362 . In warmer water a rate of 0.25 lb/acre of the primary powder yielded 88% control, while this same rate in cool weather yielded only 4% reduction . A rate of 0.5 lb/acre of the primary powder was needed to obtain 94% control of larvae in cool weather. J Clin Invest, 1985 Sep, 76(3), 970 - 7 Antigen-induced monocyte procoagulant activity . Requirement for antigen presentation and histocompatibility leukocyte antigen-DR molecules; Schwartz BS; The present study explores the interactions between lymphocytes and monocytes that are required for expression of procoagulant activity (PCA) by monocytes in response to purified protein derivative of the tubercle bacillus (PPD) or tularemia antigen . The PCA response was antigen specific: peripheral blood mononuclear cells (PBM) from donors sensitive to PPD or tularemia showed an increase in PCA only in response to the sensitizing antigen . The PCA was tissue factorlike in that Factors VII and X were required for expression of the activity, whereas Factor VIII was not . Maximum PCA developed only after 36 to 72 h . Fractionation of PBM into lymphocytes and monocytes after antigenic stimulation demonstrated that greater than 90% of the PCA was associated with monocytes . Isolated monocytes or lymphocytes incubated with sensitizing antigen had the same PCA as control cells . Purified lymphocytes that had been pulsed with antigen were unable to elicit a PCA response from monocytes to which they were added . However, adherent monocytes incubated with antigen, then washed free of unbound protein, were able to trigger lymphocytes to become stimulatory for PCA toward responding monocytes . The development of antigen-specific PCA in PBM could be blocked by including a monoclonal antibody to HLA-DR antigen in the incubation . The antibody had no effect on the clotting assay, on preformed PCA, cell viability, or on stimulatory antigen itself . These results indicate that elaboration of PCA by mononuclear cells may be an intrinsic part of the classical immune response to antigen, and may explain the presence of fibrin in immune lesions, as well as the occurrence of thrombotic complications in many immune disorders. J Clin Invest, 1985 Sep, 76(3), 1279 - 82 A human T cell clone that mediates the monocyte procoagulant response to specific sensitizing antigen; Schwartz BS et al.; A panel of human purified protein derivative of the tubercle bacillus (PPD)-reactive T cell clones was derived by cloning out of soft agar followed by cultivation on inactivated feeder cells in the presence of interleukin-2 . 1 of 4 clones tested was able to mediate an increase in monocyte procoagulant activity (PCA) in response to PPD . All four clones had identical surface marker phenotypes (T4+, T8-) and proliferated in response to antigen . The reactive T cell clone possessed no PCA of its own, but upon being presented with PPD was able to instruct monocytes to increase their expression of PCA . Antigen presentation could be performed only by autologous monocytes; allogeneic monocytes from donors unrelated to the donor of the reactive clone could not present antigen to cells of the clone in a way that would initiate the procoagulant response . Cells of the reactive clone did not mediate increased monocyte PCA in response to Candida, even though peripheral blood mononuclear cells from the donor demonstrated increased PCA to both Candida and PPD . Thus, the PCA response to specific antigen can be mediated by a single clone of cells that shows specificity in the recognition of both antigen and antigen presenting cell. FEBS Lett, 1985 Aug 19, 188(1), 91 - 5 Replacement of Mn(III) with Cu(II) in Bacillus stearothermophilus superoxide dismutase . Similarity of the active site to the zinc site of copper/zinc superoxide dismutase; Bannister JV et al.; Copper(II) was substituted for manganese(III) in Bacillus stearothermophilus Mn-superoxide dismutase . The (EPR) spectrum of the Cu(II) is distinctly rhombic, overlapping that of Cu(II) replacing zinc in copper/zinc superoxide dismutase . The copper-cyanide complex of the bacterial enzyme gives an EPR spectrum nearly identical to the cyanide-copper complex of the Cu/Zn enzyme, indicating three nitrogens as metal ligands . These results support the suggestion that metal coordination in B . stearothermophilus Mn-dismutase is a tetrahedral arrangement of three imidazoles and a carboxylate group and indicate copper as a good spectroscopic probe for studying the active site of Mn- and Fe-superoxide dismutases. J Biol Chem, 1985 Aug 15, 260(17), 9784 - 92 In vivo mobility of fatty acid end groups of Bacillus thuringiensis plasma membrane lipids during growth and sporulation; Bechtel DB et al.; The mobility of 13C specifically labeled branched chain end groups of iso-even fatty acids in intact, live Bacillus thuringiensis cells was studied by 13C nuclear magnetic resonance spectroscopy . This study apparently represents the first direct observation of branched chain carbon atoms in living cells . End groups were labeled using DL-{beta, delta, delta'-13C}valine as a precursor chain initiator for iso-even fatty acid synthesis after using L-{delta, delta'-14C}L-valine to determine optimal conditions for labeling of the membrane fatty acid end groups . Cell survival in the NMR was determined for various lengths of time at 28 and 39 degrees C . Subsequently, 13C-labeled vegetative cells, sporulating cells (three stages of development), and purified mature spores were analyzed by 13C NMR using corresponding unlabeled cells as controls . Spin lattice relaxation times (T1) were obtained for the enriched iso-branched region at 23.3 ppm and for the natural abundance peak for the glycerol backbone (carbons 1 and 3) of the membrane lipids at 61.7 ppm . The T1 of the glycerol carbons (0.08 s) did not change significantly with stage of development or temperature . The T1 of the iso-even enriched end group changed dramatically from vegetative cells (0.70s) to sporulating cells (0.28 s) at 28 degrees C . A decrease in the T1 was also observed at 39 degrees C from 0.91 s for vegetative cells to 0.54 s for sporulating cells . Accompanying the reduced mobility indicated by the T1 values, there was a general decline in the signal-to-noise ratios of identically acquired spectra as sporulation continued which culminated in the lack of discernible plasma membrane lipid resonances in purified mature spores . The progressive loss of signal appeared to have resulted from a continuous decline in the fraction of plasma membrane fatty acids with sufficient mobility to give signals above background. Biochim Biophys Acta, 1985 Aug 7, 808(3), 448 - 54 Synthetic abilities of Euglena chloroplasts in darkness; Gomez-Silva B et al.; Protein synthesis, normally a light-dependent process in isolated mature chloroplasts of Euglena gracilis var . bacillaris will take place in darkness if ATP and Mg2+ (ATP/Mg) are supplied . Either 5 or 10 mM ATP plus 15 mM MgCl2 are optimal and rates equal to those in the light can be obtained . Since ATP and Mg2+ are not stoichiometrically related, and since the optimal Mg2+ concentration is similar to that which stabilizes chloroplast ribosomes in vitro, it is suggested that the chloroplast is freely permeable to Mg2+ under these conditions . Protein synthesis under these conditions is not inhibited appreciably by DCMU, FCCP, cycloheximide, or by the addition of ribonuclease, but is highly sensitive to chloramphenicol . Carbon dioxide fixation is also a light-dependent process in isolated mature chloroplasts from Euglena, but addition of ATP (5 mM) and fructose bisphosphate (5 mM) plus aldolase (1.0 unit/ml) (fructose-1,6-bisphosphate/aldolase) yields CO2 fixation rates in darkness that are 43% of those normally obtained in the light . Mg2+ higher than 1.0 mM (e.g., 16 mM) is somewhat inhibitory . Chlorophyll synthesis from 5-aminolevulinate in 36 h developing chloroplasts from Euglena is also light-dependent, but addition of ATP/Mg and fructose-1,6-bis-phosphate/aldolase in darkness brings about the accumulation of a compound having the same RF on chromatography as protochlorophyllide from Barley; a subsequent brief illumination of the chloroplasts converts this compound to a compound with the RF of chlorophyll . Thus Euglena chloroplasts supplied with appropriate additions can carry out protein synthesis, carbon dioxide fixation and most of chlorophyll synthesis in darkness . This versatility is appropriate in photosynthetic organelles isolated from photo-organotrophic cells. Biochem J, 1985 Aug 1, 229(3), 791 - 7 The production and molecular properties of the zinc beta-lactamase of Pseudomonas maltophilia IID 1275; Bicknell R et al.; The production and purification of a tetrameric zinc beta-lactamase from Pseudomonas maltophilia IID 1275 were greatly improved . Three charge variants were isolated by chromatofocusing . The subunits each contain two atomic proportions of zinc and (in two of the variants) one residue of cysteine . The thiol group is not required for activity, nor does it appear to bind to the metal . Replacement of zinc by cobalt, cadmium or nickel takes place at a measurable rate, and gives enzymes that are less active than the zinc enzyme . The properties of this enzyme differ from those of the other known zinc beta-lactamase, beta-lactamase II from Bacillus cereus . The amino acid sequence of the N-terminal 32 residues was determined; there is no similarity to the N-terminal sequences of other beta-lactamases. Ann Allergy, 1985 Aug, 55(2), 167 - 9 Cromolyn prevents bronchospastic attacks caused by Bacillus Calmette Geurin immunotherapy for malignant melanoma patient; Wishnitzer R et al.; Bacillus Calmette Geurin (BCG) skin immunization is an adjuvant therapy for malignant melanoma patients . Adverse reactions to BCG skin immunization therapy are not uncommon and usually present as fever, influenza-like syndrome, pruritus skin rash, ulcers at the injection site, regional lymphadenopathy, and liver dysfunction . To our knowledge, a bronchospastic BCG immunotherapy-related attack has not yet been reported . We present a case of a patient with malignant melanoma who developed an unusual reaction during BCG immunotherapy . He developed asthma-like attacks after treatment, which could be prevented by inhalation of Cromolyn prior to and after therapy. J Bacteriol, 1985 Aug, 163(2), 738 - 47 Purification of the larvicidal toxin of Bacillus sphaericus and evidence for high-molecular-weight precursors; Baumann P et al.; Crystals were purified from spore-crystal complexes of Bacillus sphaericus 2362 by disruption in a French pressure cell followed by centrifugation through 48% (wt/vol) NaBr . Crystals from such preparations had a 50% lethal concentration of 6 ng of protein per ml for the larvae of the mosquito Culex pipiens . When subjected to polyacrylamide gel electrophoresis under denaturing conditions, the proteins in B . sphaericus crystals migrated in positions corresponding to 43, 63, 98, 110, and 125 kilodaltons (kDa); solubilization of the crystal at pH 12 with NaOH eliminated all but the bands at 43 and 63 kDa . Since NaOH-solubilized preparations were toxic to mosquito larvae, these proteins were purified to electrophoretic homogeneity and antiserum was obtained to each . Analysis of the two purified proteins indicated that the 43-kDa protein was toxic to mosquito larvae (50% lethal concentration, 35 ng of protein per ml), whereas the 63-kDa protein was not . Further differences between them were their amino acid compositions, their lack of immunological cross-reactivity, their opposite net charges at pH 7.5, and their susceptibility to digestion by larval midgut proteases (the 63-kDa protein was highly susceptible, whereas the 43-kDa protein was not) . The sequence of the 40 N-terminal residues of the 43-kDa protein was determined and found to contain a high percentage of hydrophobic amino acids . The sequence of the 63-kDa protein could not be determined, since it had multiple N termini . By electrophoretically separating the crystal proteins and then electroblotting onto nitrocellulose paper and visualizing the bands with antisera to the 43- and 63-kDa proteins in conjunction with an immunoblot assay, it was found that the high-molecular-mass crystal proteins (98 to 125 kDa) contained antigenic determinants of both proteins . These results suggested that the lower-molecular-weight crystal proteins detected in polyacrylamide gels after electrophoresis under denaturing conditions were derivatives of one or more of the higher-molecular-weight crystal proteins . In vivo studies of the products of crystal degradation by larvae of Culex pipiens indicated that the high-molecular-weight proteins and the 63-kDa antigenic determinants were rapidly degraded and that a 40-kDa protein related to the 43-kDa toxin persisted for the duration of the experiment (4 h) . Some of the studies performed with B.sphaericus 2362 were extended to strains 1593, 1691, and 2297 of this species with results which indicated a high degree of similarity between the crystal proteins of all these larvicidal strains. Int J Radiat Biol Relat Stud Phys Chem Med, 1985 Aug, 48(2), 223 - 33 Differential modification of oxic and anoxic components of radiation damage in Bacillus megaterium spores by caffeine; Kesavan PC et al.; Studies were carried out on the effect of caffeine on the X-irradiation sensitivity of B . megaterium spores with the following results: Caffeine exerts a concentration-dependent modifying action on oxygen-dependent components of X-ray-induced damage in B . megaterium spore suspensions causing an 'over-O2 effect' at about 1 X 10(-4) mol dm-3, and as the concentration is increased to 1 X 10(-3) mol dm-3 or above, a small but consistent protection is seen . In the absence of O2, at a wide range of concentrations (8.5 X 10(-5) to 1 X 10(-1) mol dm-3), caffeine enhances the inactivation constant, k, from 1.17 to about 1.50 kGy-1 . Both ethanol and t-butanol (5 X 10(-2) mol dm-3) remove the 'over O2-effect' produced by 1.10(-4) mol dm-3 caffeine in O2; such an effect, however, is not accompanied by reduction in the H2O2 concentrations in the spore suspensions . Ethanol prevents caffeine-induced anoxic sensitization, as well as H2O2 buildup . t-BuOH has no influence on either the low dose part of the log fraction survival curve or on the H2O2 yield in the spore suspensions . Caffeine reacts with radiation-induced eaq and .OH with rate constants of 1.5 X 10(10) and 6.9 X 10(9) dm3 mol-1s-1, respectively. J Gen Microbiol, 1985 Aug, 131 ( Pt 8), 2007 - 11 The free lipids of Mycobacterium leprae harvested from experimentally infected nine-banded armadillos; Minnikin DE et al.; The free lipids of a sample of Mycobacterium leprae were extracted by a procedure designed to produce separate non-polar and polar fractions . The composition of these lipids was analysed semi-quantitatively by five special thin-layer chromatographic systems covering the total range of mycobacterial lipid polarities . In order of increasing polarity, the major lipids were dimycocerosates of phthiocerol A, phthiocerol B and phthiodiolone A, glycosyl phenolphthiocerol dimycocerosates and phospholipids, including monoacylphosphatidylinositol di- and pentamannosides . The diacylated forms of these latter lipids, found in most mycobacteria, were not present . The composition of the free lipids of the leprosy bacillus, surveyed over the total polarity range for the first time, showed that the patterns were particularly related to those of Mycobacterium bovis, Mycobacterium kansasii and Mycobacterium marinum. Nature, 1985 Aug 1-7, 316(6027), 450 - 2 Genes for the major protein antigens of the leprosy parasite Mycobacterium leprae; Young RA et al.; Leprosy, a chronic infectious disease afflicting between 10 and 15 million people, is caused by the obligate intracellular parasite Mycobacterium leprae . Although M . leprae was the first identified bacterial pathogen of man, basic biochemical, immunological, diagnostic and therapeutic investigations have been severely limited because it remains one of the few human pathogens that have not been cultured in vitro . An M . leprae recombinant DNA expression library was constructed to provide a source of genes encoding proteins relevant for such studies . Monoclonal antibodies directed against M . leprae specific antigens have been used to isolate the genes encoding the five most immunogenic protein antigens of the leprosy bacillus . We report here that M . leprae specific epitopes recognized by all of 13 monoclonal antibodies tested were produced by recombinant phage in Escherichia coli. Infect Immun, 1985 Aug, 49(2), 371 - 7 Immunochemical characterization of a protein associated with Mycobacterium leprae cell wall; Gillis TP et al.; A panel of nine monoclonal antibodies to Mycobacterium leprae were used to characterize a protein antigen of the bacillus . Two monoclonal antibodies (IVD8 and IIIE9) were specific for M . leprae and reacted with an epitope (CWPa) present on a protein molecule associated with the cell wall fraction of M . leprae . This protein, designated cell wall-associated protein (CWP), lost its immunoreactivity upon treatment with trypsin and had an apparent molecular weight of 65,000, though additional lower-molecular-weight forms of the protein were observed by immunoblotting . Four other cross-reactive epitopes (CWPb, CWPc, CWPd, and CWPe) were defined on the same molecule using seven independent monoclonal antibodies . Therefore, M . leprae possesses a trypsin-sensitive, heat-stable protein associated with the cell wall which contains at least one species-specific and four cross-reactive antigenic determinants. Cell Immunol, 1985 Aug, 94(1), 265 - 75 Antigens associated with the activation of murine mononuclear phagocytes in vivo: differential expression of lymphocyte function-associated antigen in the several stages of development; Strassmann G et al.; Two well-characterized antigens {Mac-1 and lymphocyte-function-associated antigen (LFA-1)}, expressed on a variety of leukocytes, are members of a family of surface proteins associated with multiple recognition functions . To analyze expression of these proteins during macrophage development, we utilized both radioimmunoassay and flow cytometry . As previously reported, Mac-1 is expressed on murine macrophages in all stages of development . We found LFA-1 to be present on murine mononuclear phagocytes but only in certain stages of their development . Specifically, we found LFA-1 was expressed on murine tissue macrophages but only on those activated in vivo by bacillus Calmette Guerin (BCG) or, to a lesser extent, primed by pyran copolymer . Although LFA-1 was absent on inflammatory (responsive) and resident tissue macrophages it was also present on blood-borne monocytes . Activated macrophages also selectively expressed the H-11 and Ly-6 antigens . Thus, these data indicate that LFA-1 is selectively expressed on mononuclear phagocytes of the tissues but only on those in the primed and activated stages of development. Arch Biochem Biophys, 1985 Aug 1, 240(2), 757 - 67 Chemical modification of liquefying alpha-amylase: role of tyrosine residues at its active center; Kochhar S et al.; Liquefying alpha-amylase from Bacillus amyloliquefaciens was inactivated by treatment with tetranitromethane and N-acetylimidazole . The loss of activity occurred with modification of five tyrosine residues . Preincubation of the enzyme with either the substrate or the competitive inhibitor at saturating levels provided complete protection against inactivation . However, the presence of substrate/inhibitor in the reaction mixture protected only two of the five modifiable tyrosine residues, suggesting the involvement of only two tyrosine residues at the active center . This was confirmed when hydroxylamine treatment of the acetylated enzyme fully restored the enzymatic activity . Both nitration and acetylation increased the apparent Km of the enzyme for soluble starch, which indicated that the tyrosine residues are involved in substrate binding . Reduction of nitrotyrosine residues to aminotyrosine residues failed to restore the enzymatic activity . So, the loss of activity on modification of tyrosine residues was ascribed to conformational perturbances and not simply to the changes in the ionic character of tyrosine residues. J Biochem (Tokyo), 1985 Aug, 98(2), 333 - 9 The specific modification of histidyl residues of inorganic pyrophosphatase from Bacillus stearothermophilus by photooxidation; Shiroya Y et al.; Photooxidation of inorganic pyrophosphatase {pyrophosphate phosphohydrolase EC 3.6.1.1} from Bacillus stearothermophilus in the presence of rose bengal resulted in rapid loss of enzymatic activity . The pH profile of the inactivation rate by the photooxidation showed an inflection point around pH 6.8, suggesting the involvement of histidyl residues in the inactivation . Amino acid analysis revealed that the loss of enzymatic activity was accompanied by the destruction of 3 histidyl residues per molecule . The presence of Mg2+ alone afforded partial protection against the inactivation, whereas inorganic pyrophosphate, the substrate, showed almost no protective effect against inactivation . The photooxidation of inorganic pyrophosphatase altered the circular dichroism spectrum and the difference UV spectrum induced by Mg2+ in the near ultraviolet region . These results suggested that histidyl residues appear to be located at the binding site of Mg2+ and may contribute to the conformational change induced by Mg2+. Clin Exp Immunol, 1985 Aug, 61(2), 450 - 8 Specific cytotoxicity of human lymphocyte subpopulations defined by bacterial adherence; Spear GT et al.; It has been shown that lymphocytes can be subdivided into subpopulations based on their binding of bacteria . Monolayers of immobilized and fixed bacteria were used here to separate T cells into BA-T1T2, adherent to Escherichia coli-2 (EC-2+) and BA-T3T4, non-adherent to this strain of bacteria (Ec-2-) (our denomination) . The cells were activated in mixed lymphocyte cultures (MLC) and tested for cytotoxic activity . The BA-T1T2 cells developed the same cytotoxic activity as the sham-separated T cells whereas BA-T3T4 cells did not become cytotoxic . When the T cells were separated into BA-T2 cells, adherent to Bacillus globigii (Bg+), and BA-T1T3T4, non-adherent, (Bg- cells became cytotoxic . Since BA-T1 cells, which represent 10-20% of T cells, are common to the two populations they appear to contain all T cells needed to develop the specific cytotoxicity for allogeneic cells . When the cells were first activated in MLC for 6 days and then separated by adherence to E . coli-2 or B . globigii, all cytotoxic cells were in the non-adherent fraction . We concluded that the subpopulation of T cells which are Ec-2+Bg- (less than 20%) contain all the cells required for the development of cytoxic cell function and that after activation they become Ec-2-Bg-. FEBS Lett, 1985 Jul 8, 186(2), 229 - 32 Activity and action pattern of Bacillus licheniformis alpha-amylase in aqueous ethanol; Blakeney AB et al.; A purified B . licheniformis alpha-amylase in a mixture of ethanol-aqueous buffer (1:1, v/v) retains half the activity shown in water alone . In ethanol-aqueous buffer (7:3, v/v) about 20% of the activity is retained . The pattern of oligosaccharides produced from amylose changed with ethanol concentration; in aqueous buffer the products are: DP 1 and 2, 33.7%; DP 3, 28.5%; DP 4, 4.4% and DP 5, 33.4% . Whereas in ethanol-aqueous buffer (7:3, v/v) the products are DP 1 and 2, 66.8%; DP 3, 17.3%; DP 4, 4.1% and DP 5, 11.8% . These results suggest that a change in substrate affinity at the active centre subsites is induced in the ethanol-aqueous buffer medium. Mikrobiologiia, 1985 Jul-Aug, 54(4), 683 - 4 {Biological characteristics of the variants of Bacillus thuringiensis subsp . galleriae formed during continuous culture}; Blokhina TP et al.; Bacillus thuringiensis subsp . galleriae S . variants formed during continuous cultivation differ from the parent culture in certain properties . In contrast to the parent R form, their growth in the chemostat does not yield virulent mutants which can cause their lysis on solid media . The chemostat S forms are resistant against virulent phage mutants produced when the R variants are grown under the conditions of continuous cultivation and against a virulent phage obtained from the parent culture 69-6 under the action of vancomycin . The R forms are sensitive to these phages . When the S forms are grown under the chemostat conditions, they do not revert to the R forms . The R and S forms do not differ noticeably in the character of their growth, formation of spores and crystals, and biological activity. Mikrobiologiia, 1985 Jul-Aug, 54(4), 604 - 9 {Growth and development kinetics of Bacillus thuringiensis in batch culture}; Sakharova ZV et al.; The kinetics of Bacillus thuringiensis growth and its assimilation of nutrient substances were studied under the conditions of batch cultivation in a complex medium containing yeast extract and in a chemically defined medium with amino acids . The growth of B . thuringiensis can be divided into five phases: exponential growth; decelerated growth; stationary phase when protein crystals are formed; stationary phase when spores are formed; lysis of sporangia releasing spores . The first phase may in turn be subdivided into three stages according to changes in the specific growth rate and substrate assimilation: a high specific growth rate and no glucose assimilation; an abrupt drop in mu and the beginning of intensive glucose assimilation from the medium; a new rise in the specific growth rate . As follows from the results of studying the kinetics of B . thuringiensis growth in a chemically defined medium, the above changes in the exponential growth phase are due to the fact that the culture assimilates yeast extract components in the complex medium or amino acids in the chemically defined medium during this phase, and then starts to assimilate glucose and ammonium in the following phases of growth. Antibiot Med Biotekhnol, 1985 Jul, 30(7), 490 - 5 {Exopolysaccharide formation during the cultivation of Bacillus polymyxa on different carbohydrate substrates}; Glukhova EV et al.; The effect of the carbohydrate source in the cultivation medium on the intensity of the synthesis of exopolysaccharides of two variants of Bacillus polymyxa was studied . No significant effect of the nature of the carbohydrate source on accumulation of the culture biomass was observed, while the fermentation broth viscosity, the yield of the extracellular polysaccharides and their monosaccharide composition, as well as the ratio of the neutral and acid components significantly changed . Differences in the composition and properties of the exopolysaccharides produced in the cultures of the variant strain under identical conditions were noted . Certain correlation between the exoproduct yield and the quantity of the accumulated biomass, as well as between the composition of the exopolysaccharides and viscosity of the fermentation broth was shown, which ensures a goal-oriented effect on the synthesis of the exopolysaccharide complex by variation of the medium composition. J Biochem (Tokyo), 1985 Jul, 98(1), 157 - 65 Studies on phospholipase A inhibitor in blood plasma . II . Interaction of phospholipase A inhibitor with phospholipase A and its specificity; Miwa M et al.; Non-competitive inhibition of snake venom phospholipase A2 which has been exhibited by bovine plasma phospholipase A inhibitor, a kind of lipoprotein, was not observed unless the inhibitor was preincubated with the enzyme . The inhibition seemed to be due to the formation of the enzyme-inhibitor complex, which was identified by immunoelectrophoresis . The enzyme-inhibitor interaction was observed maximally on incubation at physiological pH, but not below pH 5 . The inhibitor was inactivated by trypsin digestion and heat treatment . It suppressed the phospholipase A2 activities of rat blood plasma as well as of the snake venom and porcine pancreas, but not the enzyme activities such as those of phospholipase C of Bacillus cereus, lipase of porcine pancreas, trypsin, and papain . The inhibitor also showed the ability to decrease membrane-bound phospholipase A1 and A2 activities in intracellular organelles such as plasma membranes, mitochondria, lysosomes, and microsomes . In view of these facts, it was concluded that the plasma inhibitor is specific for phospholipase A. Biol Chem Hoppe Seyler, 1985 Jul, 366(7), 663 - 70 Stability and self-assembly of the S-layer protein of the cell wall of Bacillus stearothermophilus; Jaenicke R et al.; The surface layer of the cell envelope of Bacillus stearothermophilus consists of a regular array of protein subunits . As shown by dodecyl sulfate polyacrylamide gel-electrophoresis and ultracentrifugation, the fully solubilized S-layer protein represents a homogeneous entity with a subunit molecular mass of 115 +/- 5 kDa . Solubilization of the protein may be accomplished at acid pH, or using high concentrations of urea or guanidine X HCl . It is accompanied by (partial) denaturation, thus interfering with the characterization of the protein in its unperturbed native state . Removal of the solubilizing agent by dialysis or dilution allows the S-layer to be reassembled into two-dimensional crystalline lattices identical to those observed in intact cells . To determine the kinetics of association, optimum conditions are found to be rapid mixing with 0.1 M sodium phosphate pH 7.0, 20 degrees C, final protein concentration greater than 10 micrograms/ml . If the time course of the self-assembly is monitored by light scattering, as well as by chemical cross-linking with glutardialdehyde, multiphasic kinetics with a rapid initial phase and slow consecutive processes of higher than second-order are observed . The rapid phase may be attributed to the formation of oligomeric precursors (Mr greater than 10(6) ) . Concentration-dependent light scattering measurements give evidence for a "critical concentration" of association, suggesting that patches of 12-16 protein subunits fuse and recrystallize into the final (native) S-layer structure . Recrystallization tends to be complete. Biochem J, 1985 Jul 1, 229(1), 241 - 9 Role of uricase in the triggering of germination of Bacillus fastidiosus spores; Salas JA et al.; The likelihood that uric acid was the only compound capable of triggering germination of Bacillus fastidiosus spores was reinforced by the finding that ureidoglycollic acid, urea, NH4Cl, 2,8-dihydroxypurine and a combination of L-alanine and O-carbamoyl-D-serine were ineffective as germinants . Uric acid-triggered germination of B . fastidiosus was prevented by a range of inhibitors that also inhibited uricase activity in dormant spore extracts . O2 uptake during germination started immediately after addition of uric acid, possibly as a consequence of the oxidation of uric acid by the enzyme uricase . Germination showed a dependence on uric acid concentration, with a relatively high Km (4-5 mM) . During the first 10 min of germination of heat-activated spores there was no detectable change in the number of spore-cortex reducing groups, indicating that selective cortex hydrolysis is not involved in the trigger mechanism of germination of B . fastidiosus . On the basis of the results, a model is proposed in which re-initiation of uricase activity is the mechanism by which B . fastidiosus spores are triggered to emerge from the dormant state. Appl Environ Microbiol, 1985 Jul, 50(1), 68 - 80 Intestinal microbial flora after feeding phytohemagglutinin lectins (Phaseolus vulgaris) to rats; Banwell JG et al.; Incorporation of purified phytohemagglutinin (PHA) lectins derived from red kidney beans (Phaseolus vulgaris) in the diet of weanling rats will cause growth failure, malabsorption of nutrients, and bacterial overgrowth in the small intestine . These effects are not caused by feeding a similar quantity of PHA to germfree rats . To define the morphological and bacterial changes on the mucosal surfaces of the jejunum, ileum, and cecum in greater detail, we pair fed two groups of weanling rats isocaloric, isonitrogenous diets with or without 0.5% PHA protein . On the jejunal surfaces of control rats, the mucous layer was a confluent covering with sparsely scattered bacteria and protozoa . In PHA-treated rats, the mucous layer was thin and discontinuous, and the microvillous surface of the tissue was extensively populated by bacterial cells of two distinct morphotypes--a gram-negative rod and a gram-positive coccobacillus . In all PHA-treated animals, these bacteria formed adherent monospecific or mixed adherent microcolonies on the tissue surface . Tissue damage was observed in PHA-exposed jejunal tissue as evidenced by vesiculation of the microvillous plasma membrane and by damage to the brush border membrane . On the ileal surfaces of control rats, there was a thick mucous layer within which small numbers of bacteria and protozoa were seen . Segmented filamentous bacteria were anchored in the tissue surface . In PHA-treated rats, the ileal surface was only incompletely covered by a mucous layer, and the overlying mucosal surface was extensively covered by large numbers of protozoan cells (predominantly Hexamita muris) . Most of the ileal surfaces not covered by the mucous layer were occupied and virtually occluded by an overgrowth of these protozoan cells with occasional cells of Giardia muris and the tissue-associated segmented bacillus . In the ceca of control rats, the mucosa was incompletely covered by a discontinuous mucous layer and colonized by an unnamed Spirillum sp., other bacteria, and occasional protozoa . The cecal surfaces of PHA-treated rats retained most of their incomplete overlying mucous layer, which was heavily colonized by the same type of Spirillum sp . seen in untreated animals; intestinal crypts were colonized . These descriptive morphological studies demonstrate that exposure to purified PHA in the diet caused characteristic changes in the microbial ecology of the small intestine . The changes in microbial flora contributed to the malabsorption of nutrients in the small intestines of PHA-fed animals. Arch Intern Med, 1985 Jul, 145(7), 1212 - 6 Improved mortality in gram-negative bacillary bacteremia; McCue JD; From 1979 to 1982, the four years of this study, episodes of gram-negative bacillary bacteremia occurred in a 489-bed community teaching hospital--an increase of 15.9% . Mortality related to bacteremia was 19.4% overall and only 3.2% for the 158 episodes involving nonfatal underlying illnesses, lower figures than those reported in the past . The severity of underlying illnesses in bacteremic patients dominated all other clinical variables that were studied as prognostic factors for the outcome of the episode . The same bacteremia-related mortality was seen in patients who had empirically received (1) multiple-antibiotic regimens in which one or more drugs were active against the pathogenic organism(s), (2) either an appropriate aminoglycoside or beta-lactam antibiotic alone, or (3) both an aminoglycoside antibiotic and a beta-lactam antibiotic active against the pathogenic organism(s). Arch Biochem Biophys, 1985 Jul, 240(1), 1 - 8 Substrate-induced changes in sulfhydryl reactivity of bacterial D-amino acid transaminase; Soper TS et al.; D-Amino acid transaminase from Bacillus sphaericus strain ATCC 14577 is a dimer with eight cysteinyl residues per molecule (T.S . Soper, W.M . Jones, and J.M . Manning (1979) J . Biol . Chem . 254, 10,901-10,905) . The reaction of the cysteinyl residues with a variety of sulfhydryl reagents has been explored to gain insight into the physical environments around these cysteinyl residues in the absence or the presence of substrates . The native enzyme, in the pyridoxal-P conformation, appears to be a symmetrical dimer, whose SH groups react in pairs with anionic reagents such as 5,5'-dithiobis(2-nitrobenzoic acid) or the halo acids . Two SH groups react with either reagent without altering enzymatic activity . Two additional SH groups react with DTNB with loss of catalytic activity . Positively charged reagents such as beta-bromoethylamine are much more effective in inactivating the pyridoxal-P conformation of the enzyme with almost five of the eight SH groups reacting and this results in a significant loss in catalytic activity . The neutral reagent dithiodipyridine is able to detect some asymmetry in the pyridoxal-P conformation . Upon addition of a D-amino acid substrate, the enzyme is transformed into the pyridoxamine-P conformation . This conformation is much more reactive with anionic reagents and much less reactive with cationic reagents, suggesting that there is a significant change in the net charge around one of the SH groups in the pyridoxamine-P conformation . Also, titration with DTNB indicates that the enzyme is a much more asymmetric dimmer in the pyridoxamine-P conformation than in the pyridoxal-P conformation . Thus, upon binding of a D-amino acid substrate, D-amino acid transaminase is transformed into the pyridoxamine-P conformation . This results in a significant change in the environment of four of the sulfhydryl groups of the enzyme . We conclude that the enzyme is transformed from a symmetrical dimer into an asymmetrical dimer and that the net charge of one of the pairs of cysteinyl groups is changed from a net negative charge into a net positive charge . These results suggest that there is a significant conformational change that occurs during the transition from the pyridoxal-P into the pyridoxamine-P form of this transaminase. Vet Med (Praha), 1985 Jul, 30(7), 433 - 41 {The activity of Bacillus thuringiensis var . israelensis against black fly larvae under field conditions in South Bohemia}; Olejnicek J et al.; The effectiveness of Bacillus thuringiensis var . israelensis spore suspension against black fly larvae was tested in two regulated brooks in the Ceske Budejovice district, South Bohemia . The sites under study were small, regulated, paved brooks of trapezoidal profile, with vegetation of different density, and with stream flow rates ranging from 40 to 90 cm/sec . Lyophilized spores (produced by Roger Belon) at a final concentration of 1 X 10(5) spores per ml were used . The spore efficacy was determined in both natural and artificial substrates in the field and under laboratory conditions, on the larvae collected at the control sites . A 90-100% mortality of the larvae was recorded in all parts of the brooks examined both in the natural samples studied in laboratory and in those kept on artificial substrates . In the area of fading spore suspension action, a higher percentage of the last-instar larvae was found in larval population which may be ascribed, in agreement with other authors, to an increased susceptibility of younger larvae to the preparation . The results of the tests demonstrated that Bacillus thuringiensis var . israelensis is an effective agent to control the black fly species occurring in Central Europe and may be of value mainly in the control of some species of veterinary importance overpopulated in regulated brooks. Scand J Immunol, 1985 Jul, 22(1), 93 - 8 Characterization of the specificity of monoclonal antibodies against Mycobacterium tuberculosis by crossed immunoelectrophoresis; Harboe M et al.; The specificity of monoclonal antibodies against Mycobacterium tuberculosis was determined by crossed immunoelectrophoresis . Radiolabelled, non-precipitating monoclonal antibody was incorporated in the top gel together with unlabelled polyvalent, precipitating anti-bacillus Calmette-Guerin (BCG) antibody . Inclusion of labelled antibody in individual precipitate lines was demonstrated by autoradiography . Each monoclonal antibody was localized in a single precipitate line: antibody TB73 reacted with BCG antigen 56; TB71 and TB72 reacted with BCG antigen 78; and TB78 reacted with BCG antigen 82 . The technique permits precise determination of the specificity of monoclonal antibodies at the level of reactivity with native immunogenic components of complex microorganisms without prior antigen purification. J Bacteriol, 1985 Jul, 163(1), 401 - 6 Nucleotide sequence of the Bacillus stearothermophilus alpha-amylase gene; Nakajima R et al.; The nucleotide sequence of the Bacillus stearothermophilus alpha-amylase gene and its flanking regions was determined . An open reading frame was found, comprising a total of 1,647 base pairs (549 amino acids) and starting from a GUG codon as methionine . It was shown by NH2-terminal amino acid sequence analysis that the extracellular amylase consisted of 515 amino acid residues, which corresponded to a molecular weight of 58,779 . Thus the NH2-terminal portion of the gene encodes 34 amino acid residues as a signal peptide . The amino acid sequence deduced from the alpha-amylase gene was fairly homologous (61%) with that of another thermostable amylase from Bacillus amyloliquefaciens. Indian J Lepr, 1985 Jul-Sep, 57(3), 519 - 23 Levels of the third component of complement in Mycobacterium leprae infected intact and immune compromised mice; Vaishnavi C et al.; Normal and immunosuppressed mice were infected with Mycobacterium leprae and the bacillary counts were made from the footpads at 3, 6 and 9 months post inoculation . A decrease in the serum C3 level was observed in the infected groups of animals compared to controls. Indian J Lepr, 1985 Jul-Sep, 57(3), 507 - 13 Primary dapsone resistance in leprosy; Vaishnavi C et al.; 20 carefully selected untreated patients of bacilliferous leprosy were investigated for primary dapsone resistance by foot pad inoculation . Mice were fed on 0.001 g% and 0.01 g% of dapsone during the period of study . Mice in the control group were given normal rodent feed only . Animals were sacrificed from 6 month onwards at 6 week intervals upto 9 months . In two animals the growth of M . leprae was not inhibited by 0.001 g% concentration of dapsone in diet, but was completely inhibited by 0.01 g% of dapsone. Biochimie, 1985 Jul-Aug, 67(7-8), 737 - 43 Engineering of tyrosyl tRNA synthetase; Bedouelle H et al.; The gene encoding the enzyme tyrosyl tRNA synthetase from Bacillus stearothermophilus has been systematically altered using synthetic oligonucleotides as mutagens . The construction of mutations has been facilitated by using strains of bacteria defective in mismatch repair and also by utilising a genetic marker in the M13 strain (such as an amber mutation, or an EcoK or EcoB site) which allows selection for the progeny of M13 replication derived from the minus (mutagenized) strand . Several mutations have been constructed in the ATP binding site to elucidate the roles of individual residues in catalysis and substrate binding and it has even been possible to construct mutants which have improved affinity for ATP . Mutations in various surface lysine and arginine residues have allowed us to identify potential contacts with the tRNA, and indicate that a cluster of basic residues close to the C-terminus of the enzyme probably makes important interactions with the tRNA. Mikrobiologiia, 1985 Jul-Aug, 54(4), 616 - 20 {Determination, by the electro-optical method, of the number of undamaged bacterial cells after extreme exposures}; Brezgunov VN et al.; The results of electro-optical measurements can be used to determine rapidly how the number of intact cells decreases before and after the action of certain extreme factors . The experiments were conducted with suspensions of Escherichia coli and Bacillus thuringiensis cells subjected to the action of heat, toluene, ethanol, and freezing . Electro-optical measurements can make it possible to assay the relative number of cells with both lethal and sublethal damages. Arch Otolaryngol, 1985 Jul, 111(7), 465 - 8 Immunomodulation of nodal lymphocytes in head and neck cancer; Schuller DE et al.; The purpose of this study was to evaluate whether the interaction of human neck node lymphocytes with primary tumor cells could be modulated with the administration of bacillus Calmette-Guerin (BCG) or N-acetylmuramyldipeptide (MDP) . The procedures involve the clonogenic assay applied towards studying the interactions of neck node lymphocytes with the tumor-stem cell population . It has been previously demonstrated that a dynamic interaction occurs between neck nodal lymphocytes and tumor . Bacillus Calmette-Guerin or MDP was incubated for three days with lymphocytes from neck nodes, after which the human squamous tumor cells from the patient were exposed continuously to the resulting lymphokines in soft agar . Ninety percent of the cases studied exhibit inhibition of tumor cell proliferation with BCG- or MDP-treated lymphocytes . The data suggest both BCG and MDP may enhance regional nodal lymphocyte tumor inhibition. South Med J, 1985 Jul, 78(7), 869 - 71 Failure of chloramphenicol and cefotaxime therapy in Klebsiella meningitis: possible role of antibiotic antagonism; Brown TH et al.; We have described a patient with Klebsiella pneumoniae meningitis who was treated with cefotaxime and chloramphenicol concomitantly, and whose slow initial resolution and subsequent relapse plus in vitro evidence of antagonism of cefotaxime appear to indicate that chloramphenicol interfered with the activity of the cephalosporin . Thus, concomitant use of chloramphenicol should probably be avoided or used advisedly in adults with gram-negative bacillary meningitis susceptible to a third generation cephalosporin. J Urol, 1985 Jul, 134(1), 48 - 53 Intravesical bacillus Calmette-Guerin therapy for superficial bladder cancer: effect of bacillus Calmette-Guerin viability on treatment results; Kelley DR et al.; We treated 40 patients with superficial bladder cancer via intravesical bacillus Calmette-Guerin for 1) prophylaxis against tumor recurrence, 2) residual carcinoma or 3) flat carcinoma in situ . A single course of intravesical bacillus Calmette-Guerin therapy was successful in 6 of 11 patients (55 per cent) treated for residual carcinoma and 6 of 12 (50 per cent) treated for carcinoma in situ . Of 17 patients receiving a single course of bacillus Calmette-Guerin for prophylaxis 11 remained free of tumor during short-term followup . A second course of therapy was administered to failures in each treatment category, which resulted in favorable responses in 5 of