Bacillus

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 6 patients treated for prophylaxis, 2 of 5 treated for residual tumor and 3 of 6 treated for carcinoma in situ . Over-all complete responses were achieved in 16 of 17 patients (94 per cent) treated for prophylaxis, 8 of 11 (73 per cent) for residual carcinoma and 8 of 12 (66 per cent) for carcinoma in situ, with a mean followup from the final treatment of 9.3, 12.3 and 7.9 months, respectively . Favorable results occurred more frequently among patients who exhibited a granulomatous inflammatory response in the bladder and delayed hypersensitivity skin test response to purified protein derivative . Marked variability in viability of bacillus Calmette-Guerin organisms was observed among different lots of bacillus Calmette-Guerin, and a direct relationship was observed between bacillus Calmette-Guerin vaccine viability and therapeutic efficacy . Most patients who failed initial therapy with a low viability lot of bacillus Calmette-Guerin responded favorably to re-treatment with a higher viability lot . The results suggest that the level of viability of each lot of bacillus Calmette-Guerin vaccine should be verified before clinical use.

J Urol, 1985 Jul, 134(1), 40 - 7
Bacillus Calmette-Guerin immunotherapy for bladder cancer; Lamm DL; Bacillus Calmette-Guerin immunotherapy has been found by a number of investigators to be effective in the treatment and prevention of superficial bladder cancer . While the optimal protocol for bacillus Calmette-Guerin remains to be determined, experience with 92 randomized and 30 nonrandomized (high risk) patients followed for up to 5 years provides information that may improve future protocols . Side effects of bacillus Calmette-Guerin are observed to increase with increasing frequency and duration of treatment . The protection from tumor recurrence has persisted: only 6 of 30 patients (20 per cent) treated with bacillus Calmette-Guerin have had recurrent tumor compared to 14 of 27 controls (52 per cent, p equals 0.008, chi-square test), and mean time to recurrence increased from 24 to 48 months (p less than 0.005, Savage) . Skin test reactivity to purified protein derivative is particularly useful in predicting response to bacillus Calmette-Guerin immunotherapy . Currently, 60 patients have been randomized to receive bacillus Calmette-Guerin immunotherapy and only 1 of 22 patients (4.5 per cent) in whom the purified protein derivative skin test results converted from negative to positive has had recurrent tumor, compared to 12 recurrences (32 per cent) in patients whose skin tests were positive before treatment or failed to convert following treatment (p equals 0.014, chi-square) . Seven recurrences (33 per cent) developed in 21 patients whose skin tests remained negative (p equals 0.015) and 5 recurrences (29 per cent) developed in 17 patients whose tests previously were positive (p equals 0.068, Fisher's test, not significant) . The benefit of percutaneous bacillus Calmette-Guerin is suggested by the observations that the recurrence rate in patients treated with intravesical bacillus Calmette-Guerin alone is 40 per cent, and all 7 patients whose purified protein derivative skin tests were negative continued to have negative results when percutaneous bacillus Calmette-Guerin was omitted (p equals 0.003) . Among high risk patients a marked decrease in or complete prevention of recurrent tumor was observed in 82 per cent of 22 patients treated previously with chemotherapy and 11 of 14 (78 per cent) with carcinoma in situ have had a complete response.

J Urol, 1985 Jul, 134(1), 36 - 9
The use of bacillus Calmette-Guerin in the therapy of bladder carcinoma in situ; Brosman SA; Intravesical instillations of Tice strain bacillus Calmette-Guerin were given to 33 patients with biopsy proved carcinoma in situ . An induction phase consisting of 12 weekly instillations was followed by a maintenance phase of instillations bimonthly for 3 months and then monthly for 18 months . A total of 6 patients did not complete the induction phase because of adverse reactions but 4 were rendered free of tumor and have had no recurrence . Of the remaining 27 patients 18 became free of tumor after 12 weeks of therapy (3 had recurrences during the maintenance period), 6 after 18 weeks (with 1 recurrence) and 3 after 24 weeks . Thus, 31 of 33 patients (94 per cent) were rendered free of carcinoma in situ . There were 4 recurrences in these patients (13 per cent) . The 27 patients who have remained free of disease have been followed for 1.75 to 8.5 years, with an average of 5.25 years . Side effects, principally bladder irritability, were a problem early in the study . With the use of isoniazid, nonsteroidal anti-inflammatory agents and bladder antispasmodics, the treatment has been safe and tolerated well . The study indicates that bacillus Calmette-Guerin is effective in eliminating carcinoma in situ in most patients, although prolonged treatment may be necessary . Maintenance therapy appears to be of value in reducing the incidence of recurrent tumor.

Cell Immunol, 1985 Jul, 93(2), 398 - 405
Enhanced interleukin 1 (IL-1) production mediated by mouse serum amyloid P component; Sarlo KT et al.; Purified serum amyloid P component (SAP), the major acute-phase reactant of mice, induces enhanced interleukin 1 (IL-1) production by elicited monocytes/macrophages in vitro . SAP also enhanced IL-1 elaboration by macrophages from lipopolysaccharide (LPS)-low responder mice and in the presence of polymyxin B, indicating that the small amounts of LPS present in the SAP preparation did not augment IL-1 production . Concentrations of SAP of 0.1 to 10.0 micrograms/ml enhanced IL-1 production by elicited and bacillus Calmette-Guerin (BCG)-activated peritoneal macrophages, but not by resident peritoneal macrophages . The inflammation-induced monocyte/macrophage population displayed selective binding of SAP . The mouse macrophage line P388D1, also could bind SAP and display enhanced IL-1 production in response to SAP . SAP did not bind to the macrophage cell line RAW264.7 nor did it enhance IL-1 secretion by this line . The results suggest that this acute-phase reactant has the potential to enhance inflammatory and immunological events mediated by IL-1.

Indian J Lepr, 1985 Jul-Sep, 57(3), 483 - 90
Multi-drug therapy in bacilliferous leprosy--two years experience; Kaur S et al.; The data of thirty patients who completed two years of multi-drug therapy (MDT) is analysed in regard to clinical improvement, occurrence of reactions, bacteriological response and compliance to therapy.

Pediatr Pulmonol, 1985 Jul-Aug, 1(4), 207 - 14
Alterations of airway smooth muscle cell membrane by sensitization; Souhrada M et al.; The sensitization of guinea pigs with ovalbumin and Bacillus pertussis vaccine caused an increase (P less than 0.001) in the resting membrane potential (Em) of airway smooth muscle (ASM) cells, from -61.3 +/- 0.5 mV (+/- SE) to -72.7 +/- 0.6 mV (+/- SE) . One, two, and three weeks after resensitization of sensitized animals with ovalbumin, Em further increased (P less than 0.001) to -76.2 +/- 0.2 mV (+/- SE), -77.4 +/- 0.3 mV, and -78.1 +/- 0.5 mV, respectively . Similarly, both in vivo and in vitro passive sensitization caused an increase (P less than 0.001) in Em of ASM cells to -69.5 +/- 0.3 mV and -68.5 +/- 0.4 mV, respectively . ASM preparations isolated from rabbits showed a similar increase (P less than 0.001) in Em after passive in vitro sensitization with serum from ovalbumin-sensitized rabbits . An increase in the contribution of the electrogenic Na+-pump was found to be responsible for the observed changes in Em following both active and passive sensitization . The presence of diphenhydramine (anti-H1), methysergide, indomethacin, 5,8,11,14-eicosatetraenoic acid (ETYA), FPL 55712 (a leukotriene receptor blocker), phenoxybenzamine, and disodium cromoglycate (a stabilizing agent) during passive in vitro sensitization failed to prevent an increase in the Em of ASM cells . However, incubation of normal guinea pig and rabbit ASM preparations with heated serum (60 degrees C for 2 hours) from ovalbumin-sensitized guinea pigs or rabbits partially inhibited (in guinea pigs) or completely abolished (in rabbits) such an increase.(ABSTRACT TRUNCATED AT 250 WORDS)

Biochem Biophys Res Commun, 1985 Jun 28, 129(3), 965 - 71
Bacterial lipopeptides induce ion-conducting pores in planar bilayers; Maget-Dana R et al.; Bacterial lipopeptides, known for their antibiotic activities, have been tested for their ability to interact with lipid membranes . These lipopeptides, Iturin A, Bacillomycin L and D and Peptidolipin NA present analogous structural characteristics: a heptapeptidic cycle is linked to a hydrocarbon chain . We present evidence that these lipopeptides modify the conductance of planar bilayers by forming ion-conducting pores.

Biochim Biophys Acta, 1985 Jun 14, 835(1), 92 - 7
The biosynthesis of leukotriene B4, the predominant lipoxygenase product in rabbit alveolar macrophages, is enhanced during immune activation; Hsueh W et al.; Rabbit alveolar macrophages synthesize prostaglandins in response to various stimuli . We have previously shown that prostaglandin production was decreased in immunologically activated (live Bacillus Calmette-Guerin (BCG)-injected) animals . (Hsueh, W., Lamb, R . and Gonzalez-Crussi, F . (1982) Biochim . Biophys . Acta 710, 406-414) . In the present study, we examined the lipoxygenase products of alveolar macrophages from normal and immunologically activated rabbits injected intravenously with live BCG or complete Freund's adjuvant . We found: unstimulated lung macrophages produced no detectable leukotrienes; the predominant lipoxygenase product upon stimulation was leukotriene B4; alveolar macrophages did not significantly degrade leukotriene B4 into its 20-hydroxy derivative, and the total degradation of leukotriene B4 during 90 min of incubation was minimal; the production of leukotriene B4 reached the peak at 30 min after A23187 stimulation, while zymosan caused a much slower and smaller release; following stimulation, immunologically activated lung macrophages produced more leukotriene B4 than resident macrophages . It is possible that the increased leukotriene B4 production in immunologically activated lung macrophages was related to the immunoregulatory function of this substance, such as enhancing cytotoxicity, interferon production and proliferation of suppressor-cytotoxic T cells.

Biochemistry, 1985 Jun 4, 24(12), 2822 - 6
Gramicidin K, a new linear channel-forming gramicidin from Bacillus brevis; Koeppe RE 2nd et al.; A new gramicidin has been isolated from a commercial mixture of gramicidins A, B, and C . This new molecule, designated gramicidin K, contains formyl and ethanolamine blocking groups, has a molecular weight approximately 20% higher than gramicidin A, and is strongly retained on reversed-phase liquid chromatographic columns . Gramicidin K can be resolved into two components, one of which contains tyrosine . In lipid bilayer membranes, both components form channels of considerably longer lifetime and somewhat lower conductance than gramicidin A . Gramicidin K appears to be a lipopeptide that consists of a fatty acyl chain attached to the ethanolamine of gramicidin A.

Eur J Biochem, 1985 Jun 3, 149(2), 331 - 6
Structural studies on the linkage unit between poly(N-acetylglucosamine 1-phosphate) and peptidoglycan in cell walls of Bacillus pumilus AHU 1650; Kojima N et al.; Structural studies were carried out on the polymer chains and their linkage regions in two kinds of teichoic acids, poly(N-acetylglucosamine 1-phosphate) {poly(GlcNAc-1-P)} and glycerol teichoic acid, bound to peptidoglycan in the cell walls of Bacillus pumilus AHU 1650 . The poly(GlcNAc-1-P)-glycan complex isolated from lysozyme digests of the cell walls contained mannosamine and glycerol as minor components . On the basis of proton NMR spectroscopic data and isolation of N-acetylglucosamine 4-phosphate from acid hydrolysates, the poly(GlcNAc-1-P) was shown to be a polymer in which N-acetylglucosamine 1-phosphate units are joined at C-4 of the glucosamine residues . Mild alkaline hydrolysis of the poly(GlcNAc-1-P)-glycan complex gave a mannosamine-linked glycan fragment and the acidic polymer fraction that contained glycerol residues . Mild acid treatment of the mannosamine-linked glycan fragment gave the linkage disaccharide, ManNAc(beta 1----4)GlcNAc, whereas the acidic polymer fraction was degraded by this treatment into N-acetylglucosamine 4-phosphate and a glycerol-containing fragment characterized as P-(Gro-P)7 (Gro = glycerol) . On the other hand, direct mild acid hydrolysis of the complex gave a fragment characterized as P-(Gro-P)7-ManNAc(beta 1----4)GlcNAc . These results lead to a conclusion that in the cell walls the poly(GlcNAc-1-P) chain is attached to peptidoglycan through a linkage unit, (Gro-P)7-ManNAc(beta 1----4)GlcNAc . By means of similar procedures, it was shown that the other cell wall polymer, glycerol teichoic acid, is also attached to peptidoglycan through the same disaccharide, ManNAc(beta 1----4)GlcNAc.

Eur J Biochem, 1985 Jun 3, 149(2), 363 - 73
Nucleotide sequence and high-level expression of the major Escherichia coli phosphofructokinase; Hellinga HW et al.; The gene for the major phosphofructokinase enzyme in Escherichia coli, pfkA, has been sequenced . Comparison of the amino acid sequence with other phosphofructokinases showed that this enzyme is related to the Bacillus stearothermophilus and rabbit muscle enzymes, but is different from the second, minor phosphofructokinase found in E . coli . The region which has been sequenced comprises the complete pfkA--tpi interval on the E . coli genetic map . Two other genes have been identified from the nucleotide sequence: a gene for a periplasmic sulphate-binding protein, sbp, and for a membrane-bound enzyme, CDP-diglyceride hydrolase, cdh . This establishes the complete gene arrangement in this region as pfkA-sbp-cdh-tpi . The pfkA gene has been subcloned into a high-copy-number plasmid under the control of a strong, chimaeric promoter which arose as an artefact in the construction of the plasmid gene bank from which the original pfkA recombinant was isolated . A specialised recombinant has been constructed which carries a 1.4 X 10(3)-nucleotide insert containing just the pfkA gene flanked by two HindIII recognition sites providing a simple system for the recloning of this gene into different vectors . This recombinant expresses the enzyme at high levels (40-50% of total cell protein is active, soluble phosphofructokinase) . This expression system is now being used to study the enzyme using 'reverse genetics'.

Zentralbl Bakteriol Mikrobiol Hyg {B}, 1985 Jun, 181(1-2), 139 - 50
{Determination of the killing rate of Bacillus spores by peracetic acid}; Botzenhart K et al.; The sporocidal properties of peracetic acid (PAA) at defined concentrations were characterized by determination of decrease in PAA after addition of D-glucose, human albumin and suspensions of spores; concentration of PAA, which inactivates 10(6)-10(7) spores of Bacillus cereus, B . subtilis, B . megaterium and B . licheniformis within 10-30 min; inactivation constant k, decimal reduction time D and the sporocidal index (mg PAA X min X ml-1) at that concentration . In contrast to albumin, the spore suspension caused relatively little reduction of PAA concentration (less than 5% at the concentrations used) . B . cereus, B . subtilis and B . megaterium had similar inactivation rates with k-values in the range of 0.368 and 0.541 min-1, D-values between 4.26 min and 6.26 min at 0.2 mg/ml PAA . B . lichenformis was much more resistant showing a k = 0.345 min-1 and D = 6.66 min at 3.0 mg/ml PAA . The sporocidal index of B . lichenformis was 180 mg X min X ml-1 while the three other species had sporocidal index-values of 7 mg X min X ml-1.

Zh Mikrobiol Epidemiol Immunobiol, 1985 Jun, (6), 75 - 7
{Changes in the cellular reactivity of guinea pigs sensitized with Bacillus thuringiensis}; Padalkin VP et al.; Changes in the cell-mediated responsiveness of the body under the action of different variants of B . thuringiensis have been studied in experiments on guinea pigs . The data thus obtained indicate that the development of sensitization occurs in the animals, which is manifested by the increase of the sensitivity of leukocytes to the specific allergen and by the increase of the phagocytic activity of peritoneal macrophages . The most pronounced changes in the immune responsiveness of guinea pigs have been observed after the parenteral administration of B . thuringiensis var . galleriae.

Ann Gastroenterol Hepatol (Paris), 1985 Jun, 21(3), 135 - 6
{Pseudotumoral form of hepatic tuberculosis . Apropos of a case report}; Bernard E et al.; Concerning one case of autonomous hepatic tuberculosis in a pseudo-tumoral aspect, the difficulties of making the diagnosis are brought to light . Liver biopsy is the best examination to clarify things, either performed under ultrasonic control or during surgery . This biopsy permits pathologic examination and culture of a liver fragment in Lowenstein medium . But in fact, only isolation of the tubercle bacillus at the liver level confirms presence of tuberculosis.

J Clin Hosp Pharm, 1985 Jun, 10(2), 203 - 9
A note on the contamination of eye-drops following use by hospital out-patients; Ford JL et al.; Examination of 273 eye-drops, returned to a hospital pharmacy by out-patients showed that 27% were contaminated . Common contaminants included Bacillus spp, Staphylococcus albus and Sarcina spp . Those eye-drops containing the preservatives chlorbutol or thiomersal showed above average levels of contamination, thus questioning the suitability of these preservatives for ophthalmic preparations.

Arch Biochem Biophys, 1985 Jun, 239(2), 327 - 33
Failure of an alkalophilic bacterium to synthesize ATP in response to a valinomycin-induced potassium diffusion potential at high pH; Guffanti AA et al.; Starved whole cells of the obligately alkalophilic Bacillus firmus RAB synthesize ATP upon addition of L-malate at pH 9.0 as expected of an aerobic organism that grows oxidatively on nonfermentable carbon sources at pH values as high as 11.0 . The current study was a detailed examination of the perplexing inability of such cells to exhibit ATP synthesis in response to a valinomycin-mediated potassium diffusion potential at pH 9.0 . While there were minor differences in the patterns of generation of the potential and the proton influx that accompanies its generation in the three different buffering systems employed, the magnitude of the transmembrane electro-chemical potential of protons was at least as high as pH 9.0 as at pH 7.0 . Nevertheless, a diffusion potential consistently energized ATP synthesis at pH 7.0 but not at 9.0; these findings were independent of the presence or absence of Tris or of Na+ . By contrast, the artificial electron donor ascorbate, in the presence of phenazine methosulfate, energized ATP synthesis by the starved whole cells at both pH values . The same phenomenon, i.e., efficacy of a respiration-derived potential but not of a diffusion potential at pH 9.0, was demonstrated in ADP + Pi-loaded membrane vesicles . On the other hand, electrogenic Na+-coupled solute transport could be energized by both ascorbate/phenazine and methosulfate and a diffusion potential in the vesicles at pH 9.0 . The results are discussed in connection with models of a localized path of proton flow between proton pumps and the ATP synthase.

J Gen Virol, 1985 Jun, 66 ( Pt 6), 1287 - 96
The nucleocapsid of Berne virus; Horzinek MC et al.; In Berne virus-infected cells and in gradient-purified virions two major proteins with mol . wt . of 20K and 22K were detected . The 22K species is thought to represent the main envelope polypeptide; in infectious culture media it was present in a low envelope polypeptide; in infectious culture media it was present in a low density substructure which could be quantitatively converted into slowly sedimenting material by detergent treatment . The 20K polypeptide (accounting for about 80% of the 14C-amino acid label in the virion) was phosphorylated, occurred in an intracellular substructure of higher density than the virion (rho = 1.36 g/ml in CsCl) and was the only viral protein possessing RNA-binding properties; it was recognized preferentially by heterologous animal sera in immune precipitation . The 20K species is therefore identified as the main capsid protein . Two additional polypeptides (19K and 17K) were regularly detected in extracts of infected cells; they appeared to share oligopeptides with the 20K protein and are interpreted as being proteolytic cleavage products . The nucleocapsid of Berne virus was visualized after ether treatment as a flexible bacilliform structure with conspicuous transverse striation . Demonstration of a 20K nucleocapsid protein further supports the authors' proposal that Berne virus is a representative of a new family of enveloped RNA viruses (Toroviridae).

Proc Natl Acad Sci U S A, 1985 Jun, 82(11), 3539 - 42
Alternative conformations in Escherichia coli 16S ribosomal RNA; Klein BK et al.; Partially denatured 16S rRNA from 30S ribosomes shows features of secondary structure in electron microscopy that correspond to the well accepted secondary structure model derived from chemical modification and phylogenetic data . However, a very different conformation is seen in precursor 16S rRNA sequences contained within 30S pre-rRNA transcripts: the major 5'-terminal loop is absent, and several additional quite stable large loops, symmetrically placed in the molecule, are present . Features of the alternative structure are also seen in mature 16S rRNA from Escherichia coli and from two Bacillus species when heated in certain buffers . Microscopy thus reveals specific features of alternative conformations and their relative stabilities, suggesting a possible transition during ribosome formation.

J Bacteriol, 1985 Jun, 162(3), 1120 - 5
Molecular cloning of an ornithine-activating fragment of the gramicidin S synthetase 2 gene from Bacillus brevis and its expression in Escherichia coli; Krause M et al.; Partially digested chromosomal DNA of Bacillus brevis ATCC 9999, a producer of the cyclic peptide antibiotic gramicidin S, was ligated into the BamHI site of the Escherichia coli expression vector pUR2-Bam . The ligated molecules were used to transfer E . coli to ampicillin resistance . Of 5 X 10(3) colonies tested by in situ immunoassay for a cross-reaction with antibodies against the gramicidin S synthetase 2, 6 colonies were found to be immunoreactive . A clone designated MK2, which had a 3.9-kilobase insert of B . brevis DNA, directed in E . coli under the lac promoter control the synthesis of polypeptides that were cross-reactive with the antibody to the gramicidin S synthetase 2 . Partial purification of the gene products by gel filtration revealed a major fraction with an approximate molecular weight of 140,000 and with specific ornithine-dependent ATP-32PPi and 2'-dATP-32PPi exchange activities . These unique activities of the gramicidin S synthetase 2 were not detected in the E . coli strain harboring the vector.

Cell Immunol, 1985 Jun, 93(1), 189 - 98
The effect of hydrocortisone and cyclosporin A on bacillus Calmette-Guérin epitheloid cell granulomas; Gupta SK et al.; The injection of bacillus Calmette-Guerin (BCG) intradermally into the ear of guinea pigs leads to the formation in the draining lymph node of granulomas containing epithelioid cells with rough endoplasmic reticulum (RER) and an absence of phagocytosed material . BCG granulomas in hydrocortisone- or cyclosporin A (CsA)-treated animals contain mononuclear phagocytes with no RER . In CsA-treated animals, these cells contain fragments of phagocytosed organisms . CsA was given at two doses, 25 mg/kg orally and 50 mg/kg ip . The higher dose completely suppressed the delayed hypersensitivity (DH) response to purified protein derivative (PPD) but the lower dose did not affect the level of the DH response, but had a profound effect on epithelioid cell formation . The role of lymphokines in the maturation of the monocyte/macrophage to epithelioid cells is discussed.

Int J Radiat Biol Relat Stud Phys Chem Med, 1985 Jun, 47(6), 621 - 7
The contribution of endogenous and exogenous effects to radiation-induced damage in the bacterial spore; Jacobs GP et al.; Radical scavengers such as polyethylene glycol 400 and 4000 and bovine albumin have been used to define the contribution of exogenous and endogenous effects to the gamma-radiation-induced damage in aqueous buffered suspensions of Bacillus pumilus spores . The results indicate that this damage in the bacterial spore is predominantly endogenous both in the presence of 1 atmosphere of oxygen, and in anoxia.

South Med J, 1985 Jun, 78(6), 731 - 2
Successful treatment of gram-negative bacillary meningitis with imipenem/cilastatin; Rodriguez K et al.; A patient with meningitis caused by a strain of Actinetobacter anitratus that was resistant to all commercially available antibiotics was treated with imipenem/cilastatin administered intravenously in a dose of 1 gm of imipenem every six hours . The minimal inhibitory concentration of imipenem against the isolate was less than or equal to 0.04 micrograms/ml . The patient tolerated the drug well and was cured after 12 days of therapy.

Mol Gen Mikrobiol Virusol, 1985 Jun, (6), 21 - 3
{Characteristics of Bacillus thuringiensis phages with circular permutation in the DNA molecule}; Smirnova TA et al.; Bacteriophages Tm2 and Tg27 of different origins but identical in biological properties have been compared . Physicochemical characteristics of bacteriophages have revealed the existence of end repeats and circular permutation of phage DNA . Phages Tm2 and Tg27 share the same dimensions of incapsulated DNA, differing in the sizes of phage genome and end repeats . Bacteriophage Tm2 genome is 45.2 kb . long with the end repeats containing 5.7% . The genome of Tg27 is 42.53 kb and 11.8% of end repeat . The bacteriophages relation has been confirmed by heteroduplex and restriction analysis . Tm2 and Tg27 share 84% of homology . Two regions of nonhomology are found representing a single-stranded loop and equishouldered vesicle with the sizes 2.19 kb and 5.03 kb, respectively.

J Hosp Infect, 1985 Jun, 6(2), 194 - 200
Pseudo-outbreak of Bacillus species: related to fibreoptic bronchoscopy; Goldstein B et al.; In July and August 1980, Bacillus spp . were isolated with unusual frequency from bronchial washings obtained during bronchoscopy performed with fibreoptic equipment . Clinical disease in culture-positive patients was not apparent; but Bacillus spp . were recovered from one component of the bronchoscope, the automatic suction valve . It appeared that secretions became contaminated during passage through the valve en route to a collection device . After institution of controls directed primarily at proper storage and maintenance, the number of isolations of Bacillus spp . decreased significantly . Thus, investigation revealed a site for contamination not previously described and the results suggest that in future outbreaks, cultures of the component parts may permit localization of the site of contamination which may help focus control measures.

J Biol Chem, 1985 May 25, 260(10), 6273 - 80
Delineation of a toxin-encoding segment of a Bacillus thuringiensis crystal protein gene; Schnepf HE et al.; Crystals of Bacillus thuringiensis subsp . kurstaki HD-1-Dipel contain a Mr 134,000 protoxin which can be cleaved by proteolysis to a peptide of Mr approximately 70,000; this peptide is lethal to lepidopteran larvae . We have analyzed the peptides produced by recombinant Escherichia coli strains bearing deletions and fusions of the protoxin gene in order to delineate the portion of the gene which encodes the toxic peptide . The recombinant strains produced the toxic peptide as well as larger peptides whose size was related to the length of the deleted gene . The results indicate that the amino-terminal 55% of the protoxin protein is sufficient for toxicity . While two different gene fusions to the 10th codon allowed the synthesis of toxic polypeptides, fusions to the 50th codon did not . 3' end deletions up to the 645th codon allowed synthesis of the toxic peptide whereas a deletion to the 603rd codon yielded a non-toxic peptide . Some of the 5' and 3' end alterations to the gene caused changes in the proteolytic cleavage patterns of the polypeptides synthesized by E . coli, suggesting that the alterations led to conformational changes in the proteins . The presence of different 3' end segments affected the levels of synthesis of the altered crystal proteins.

J Biol Chem, 1985 May 25, 260(10), 6264 - 72
The amino acid sequence of a crystal protein from Bacillus thuringiensis deduced from the DNA base sequence; Schnepf HE et al.; We have determined the nucleotide sequence of a 4222-base segment of DNA which contains the promoter, the coding region, and the terminator of a crystal protein gene cloned from a Bacillus thuringiensis plasmid . A sequence of 1176 amino acids encoding a Mr 133,500 peptide was deduced from the single open reading frame . This protein-coding region was analyzed for codon usage, predicted hydropathy, and predicted secondary structure . Examination of the base sequence revealed the presence of several inverted and direct repeats located in both the coding and noncoding regions . S1 nuclease mapping was used to locate the transcription termination point at a site following a potentially very stable stem-and-loop structure.

Eur J Biochem, 1985 May 2, 148(3), 539 - 43
Purification and partial characterization of the extracellular gamma-D-glutamyl-(L)meso-diaminopimelate endopeptidase I, from Bacillus sphaericus NCTC 9602; Garnier M et al.; The gamma-D-glutamyl-(L)meso-diaminopimelate endopeptidase, or endopeptidase I, from Bacillus sphaericus 9602 was purified to apparent protein homogeneity . The purification was achieved by a six-step procedure: ammonium sulfate fractionation, phenyl-Sepharose chromatography, two consecutive DEAE-Trisacryl chromatographies, chromatofocusing and Sephacryl S-200 permeation chromatography . The enzyme was purified 5000-fold with a 38% recovery of lytic activity . It is an acidic protein (pI 5.4) of hydrophobic nature . Kinetic studies have shown a Km value of 0.57 mM and an apparent Vmax of 8.3 mumol min-1 (mg enzyme)-1 with N-acetylmuramyl-L-alanyl-gamma-D-glutamyl-(L)meso-diaminopimelyl (L)-D-{14C}alanine as substrate . The enzyme was inhibited by o-phenanthroline and EDTA and was reactivated by zinc, cobalt and manganese ions; thus endopeptidase I is a metallo enzyme, probably a zinc enzyme . Moreover it is a heat-stable protein with an apparent inactivation temperature of 80 degrees C.

Eur J Biochem, 1985 May 2, 148(3), 479 - 84
Structural studies on the acidic polysaccharide of Bacillus cereus AHU 1356 cell walls; Kojima N et al.; Structural studies were carried out on the acidic polymer fraction isolated from lysozyme digests of the N-acetylated cell walls of Bacillus cereus AHU 1356 . The acidic polymer fraction contained glucosamine, galactose, rhamnose, glycerol and phosphorus in a molar ratio of 1:1:2:1:1, together with small amounts of glycopeptide components and muramic acid 6-phosphate . The hydrogen fluoride treatment led to removal of glycerol and phosphorus from the polymer without loss of other components . Results of the NaIO4 oxidation, methylation and proton magnetic resonance spectroscopy of the native and dephosphorylated preparations, in combination with data of the analysis of oligosaccharides obtained from partial hydrolysis of polysaccharide, led to the most likely structure of the repeating units of the acidic polysaccharide chain, ----4)N-acetylglucosaminyl-(alpha 1----3)rhamnosyl(alpha 1----3)galactosyl(alpha 1----4){sn-glycerol 1-phospho-2}rhamnosyl(alpha 1----.

J Biochem (Tokyo), 1985 May, 97(5), 1477 - 85
Proline iminopeptidase from Bacillus coagulans: purification and enzymatic properties; Yoshimoto T et al.; Proline iminopeptidase {EC 3.4.11.5} was purified about 2,700-fold from cell-free extract of Bacillus coagulans by a series of column chromatographies on DEAE-Toyopearl, PCMB-T-Sepharose, and hydroxyapatite, and gel filtration on Sephadex G-150 . The purified enzyme was homogeneous as judged by disc gel electrophoresis . The enzyme was most active at pH 7.3 with Pro-beta-naphthylamide (Pro-2-NNap) as the substrate, and hydrolyzed Pro-X (X = amino acid including proline, peptide, amide, and arylamide) bonds when the proline residue was at the amino terminus . Pro-D-amino acid bonds were also susceptible to the enzyme . The enzyme was completely inhibited by p-chloromercuribenzoate (PCMB) and partially by proline but not by metal chelators, diisopropylphosphorofluoridate (DFP), or phenylmethanesulfonyl fluoride (PMSF) . The enzyme inactivated with PCMB was reactivated by incubation with 2-mercaptoethanol . These results and the chromatographic profile on PCMB-T-Sepharose suggest that the enzyme is a sulfhydryl enzyme . The isoelectric point of the enzyme was 4.0, and the molecular weight of the enzyme was estimated to be 40,000 by gel filtration on Sephadex G-100 and 35,000 by sodium dodecyl sulfate (SDS) gel electrophoresis, indicating that the enzyme exists as a monomer.

J Urol, 1985 May, 133(5), 786 - 8
Monitoring intravesical bacillus Calmette-Guerin treatment of bladder carcinoma with flow cytometry; Staiano-Coico L et al.; Flow cytometry of bladder irrigation specimens was studied in 22 patients with low stage bladder carcinoma who were treated by transurethral resection of visible tumor followed in 3 to 5 weeks by a course of intravesical bacillus Calmette-Guerin . The most informative examinations were just before the first bacillus Calmette-Guerin treatment, 6 weeks after completing a 6-week course of treatment (3 months) and at 9 months . Of the patients 10 had recurrent tumors after therapy; recurrence was anticipated correctly by flow cytometry at the 12-week followup examination in 6 of the 10 patients and suspected in another . Of 12 patients who remained clinically free of disease for a minimum of 15 months after bacillus Calmette-Guerin therapy flow cytometry identified correctly 7 at 12 weeks, while 1 had a partial response and the remaining 4 reverted to a negative status at 9 months . Of interest, only 4 of the 22 patients were free of disease by flow cytometry at the start of bacillus Calmette-Guerin treatment despite attempted ablation of the tumor by transurethral resection, suggesting that intravesical administration of bacillus Calmette-Guerin destroys existing carcinoma in situ in some cases.

J Cell Biol, 1985 May, 100(5), 1690 - 7
Localization of phosphatidylcholine in outer envelope membrane of spinach chloroplasts; Dorne AJ et al.; We have examined the effects of phospholipase C from Bacillus cereus on the extent of phospholipid hydrolysis in envelope membrane vesicles and in intact chloroplasts . When isolated envelope vesicles were incubated in presence of phospholipase C, phosphatidylcholine and phosphatidylglycerol, but not phosphatidylinositol, were totally converted into diacylglycerol if they were available to the enzyme (i.e., when the vesicles were sonicated in presence of phospholipase C) . These experiments demonstrate that phospholipase C can be used to probe the availability of phosphatidylcholine and phosphatidylglycerol in the cytosolic leaflet of the outer envelope membrane from spinach chloroplasts . When isolated, purified, intact chloroplasts were incubated with low amounts of phospholipase C (0.3 U/mg chlorophyll) under very mild conditions (12 degrees C for 1 min), greater than 80% of phosphatidylcholine molecules and almost none of phosphatidylglycerol molecules were hydrolyzed . Since we have also demonstrated, by using several different methods (phase-contrast and electron microscopy, immunochemical and electrophoretic analyses) that isolated spinach chloroplasts, and especially their outer envelope membrane, remained intact after mild treatment with phospholipase C, we can conclude that there is a marked asymmetric distribution of phospholipids across the outer envelope membrane of spinach chloroplasts . Phosphatidylcholine, the major polar lipid of the outer envelope membrane, is almost entirely accessible from the cytosolic side of the membrane and therefore is probably localized in the outer leaflet of the outer envelope bilayer . On the contrary, phosphatidylglycerol, the major polar lipid in the inner envelope membrane and the thylakoids, is probably not accessible to phospholipase C from the cytosol and therefore is probably localized mostly in the inner leaflet of the outer envelope membrane and in the other chloroplast membranes.

J Bacteriol, 1985 May, 162(2), 571 - 8
Protoplast dehydration correlated with heat resistance of bacterial spores; Nakashio S et al.; Water content of the protoplast in situ within the fully hydrated dormant bacterial spore was quantified by use of a spore in which the complex of coat and outer (pericortex) membrane was genetically defective or chemically removed, as evidenced by susceptibility of the cortex to lysozyme and by permeability of the periprotoplast integument to glucose . Water content was determined by equilibrium permeability measurement with 3H-labeled water (confirmed by gravimetric measurement) for the entire spore, with 14C-labeled glucose for the integument outside the inner (pericytoplasm) membrane, and by the difference for the protoplast . The method was applied to lysozyme-sensitive spores of Bacillus stearothermophilus, B . subtilis, B . cereus, B . thuringiensis, and B . megaterium (four types) . Comparable lysozyme-resistant spores, in which the outer membrane functioned as the primary permeability barrier to glucose, were employed as controls . Heat resistances were expressed as D100 values . Protoplast water content of the lysozyme-sensitive spore types correlated with heat resistance exponentially in two distinct clusters, with the four B . megaterium types in one alignment, and with the four other species types in another . Protoplast water contents of the B . megaterium spore types were sufficiently low (26 to 29%, based on wet protoplast weight) to account almost entirely for their lesser heat resistance . Corresponding values of the other species types were similar or higher (30 to 55%), indicating that these spores depended on factors additional to protoplast dehydration for their much greater heat resistance.

J Immunol, 1985 May, 134(5), 3432 - 8
Induction of protective immunity against Schistosoma mansoni by a non living vaccine . I . Partial characterization of antigens recognized by antibodies from mice immunized with soluble schistosome extracts; James SL et al.; A single intradermal injection of frozen and thawed schistosomula in conjunction with the bacterial adjuvant Mycobacterium bovis strain Bacille Calmette Guerin, Phipps substrain (BCG) induced significant levels of resistance to challenge Schistosoma mansoni infection in C57BL/6 mice . Immunization with the aqueous fraction remaining after 100,000 X G centrifugation of the larval lysate was also protective under these conditions, suggesting that some immunogenic determinants may not be membrane associated . Frozen-thawed cercariae and soluble components of adult worms also protected against challenge infection in these experiments . These observations indicate that soluble immunogens are present in both early and late developmental stages of the parasite, and therefore may be good candidate antigens for an immunochemically defined vaccine against schistosomiasis . Induction of humoral reactivity against soluble or membrane antigens was examined in mice protected against cercarial challenge by prior exposure to frozen-thawed larvae, soluble larval, or soluble adult antigens plus BCG . Animals that were immunized with frozen-thawed larvae produced low but significant levels of antibodies against larval surface antigens when examined by indirect immunofluorescence or by immunoprecipitation of surface-labeled schistosomula . Mice immunized with soluble antigens, however, showed negligible antibody reactivity against surface membrane antigens . Because mice immunized with soluble antigens were resistant to challenge infection, these results strongly suggest that anti-surface membrane reactivity is not required in the mechanism of protective immunity in this model . Sera from mice immunized with either total freeze-thaw larval lysate or soluble schistosome extracts all showed strong reactivity against soluble antigens, as detected by ELISA . Western blot analysis showed these antisera to react with a restricted number of high m.w . antigens that were present both in schistosomula and in adult worms . These antigens are therefore likely to play a major role in the development of resistance in this model as immunogens and/or as targets of protective immune response.

J Infect, 1985 May, 10(3), 228 - 32
Dissemination of Bacillus cereus in a maternity unit; Youngs ER et al.; During a period of 4 months, 54 of 1059 (5%) specimens received for bacterial culture from neonates in a maternity unit yielded Bacillus cereus . Of the strains isolated, 48 originated from 259 umbilical cord swabs examined . Furthermore, 88% of the isolates belonged to a single, previously undescribed serotype . Strains of the same serotype were also found to be widely distributed in the environment throughout the maternity unit . The primary source of the organism, however, could not be established.

Mol Cell Biochem, 1985 May, 67(1), 77 - 81
Induction of a cytochrome P-450-dependent fatty acid monooxygenase in Bacillus megaterium by a barbiturate analog, 1-{2-phenylbutyryl}-3-methylurea; Wen LP et al.; In previous publications from our laboratory, we reported that a soluble, cytochrome P-450-dependent fatty acid monooxygenase from Bacillus megaterium ATCC 14581 can be induced by phenobarbital and a variety of other barbiturates . The tested barbiturates showed an excellent correlation between increasing lipophilicity and increasing inducer potency (Kim BH, Fulco AJ; Biochem Biophys Res Commun 116: 843-850, 1983) . The only exception proved to be mephobarbital (N-methylphenobarbital) which, although more lipophilic than phenobarbital, is not an inducer of fatty acid monooxygenase activity . We have now found that 1-{2-phenylbutyryl}-3-methylurea (PBMU), an acylurea that can be derived from mephobarbital by hydrolytic cleavage of the barbiturate ring, is an excellent inducer of this activity . Paradoxically, the addition of mephobarbital to the bacterial growth medium containing PBMU significantly enhances the apparent potency of the acylurea to induce fatty acid monooxygenase activity as measured in cell-free extracts . When cell-free extracts of cells grown separately in PBMU or mephobarbital are mixed no enhancement of activity is seen . This finding suggests that the effect of mephobarbital is to somehow increase the efficiency of PBMU as an inducer of the P-450-dependent fatty acid monooxygenase rather than to induce an activator of this enzyme or a rate-limiting component of the monooxygenase system . Finally, both mephobarbital and PBMU induce the synthesis of total cytochrome P-450 in B . megaterium although PBMU is a much more potent P-450 inducer . For cytochrome P-450 induction, however, there is no synergistic or even additive effect when mephobarbital and PBMU are used together in the bacterial growth medium.

Am Rev Respir Dis, 1985 May, 131(5), 760 - 3
Techniques of DNA hybridization detect small numbers of mycobacteria with no cross-hybridization with non-mycobacterial respiratory organisms; Shoemaker SA et al.; The traditional methods used in identifying mycobacteria, such as acid-fast bacillus stains and culture, are often time-consuming, insensitive, and nonspecific . As part of an ongoing program to improve diagnosis and characterization of mycobacteria, we have found that deoxyribonucleic acid (DNA) hybridization techniques using isotopically labeled, single-stranded, total DNA can be used to detect as little as 10(-4) micrograms of Mycobacterium tuberculosis (MTb) DNA . This amount of DNA represents approximately 2 X 10(4) genomes . We have also shown the MTb DNA is sufficiently different from the DNA of non-mycobacterial microorganisms such that cross-hybridization with MTb DNA does not occur under the hybridization conditions we employed . We speculate that DNA hybridization techniques may allow the rapid, sensitive, and specific identification of mycobacteria.

Am J Pathol, 1985 May, 119(2), 223 - 35
Strain variation of bacillus Calmette-Guerin-induced pulmonary granuloma formation is correlated with anergy and the local production of migration inhibition factor and interleukin 1; Kobayashi K et al.; Pulmonary granulomatous inflammation was induced by the intratracheal injection of viable bacillus Calmette-Guerin (BCG) into genetically high granuloma responder (C57BL/6J and BALB/c) and low responder (CBA/J) mice with and without immunization by methylated bovine serum albumin in complete Freund's adjuvant . Significant migration inhibition factor (MIF) and interleukin 1 (IL 1) activities were detected in aqueous lung granuloma extracts prepared from high responder mice bearing BCG-induced granulomatous inflammation . Interleukin 2 activity was not detected . Very low MIF and IL 1 activities were detected in extracts from low responder mice . Furthermore, high responder, but not low responder, mice showed marked suppression of in vivo and in vitro manifestations of cell-mediated immunity to both specific and nonspecific antigens . In contrast, humoral antibody response was not affected significantly . The kinetics of anergy in granuloma-bearing mice correlated closely with the appearance of MIF and IL 1 activities in the lesions . Thus, genetically determined granuloma response to BCG and the expression of anergy in various strains of mice were well associated with in vivo release of MIF and IL 1 . These results indicate that the genetic ability or inability to mount a granulomatous inflammatory response to BCG may extend to the capacity of cells within the lesions to generate soluble mediator(s) which is also responsible for anergy in granuloma-bearing mice.

Arch Pathol Lab Med, 1985 May, 109(5), 465 - 6
Clues to the early diagnosis of Mycobacterium avium-intracellulare infection in patients with acquired immunodeficiency syndrome; Polis MA et al.; Four patients with acquired immunodeficiency syndrome with bacillemia from Mycobacterium avium-intracellulare presented with early pathologic clues of a disseminated mycobacterial infection . All had persistent fevers with negative diagnostic workups for other usual pathogens seen in patients with acquired immunodeficiency syndrome . Two patients had prolonged clearance of the bacillemia on a drug regimen of ansamycin, clofazimine, and amikacin sulfate.

J Bacteriol, 1985 May, 162(2), 682 - 92
Isolation of a Bacillus stearothermophilus mutant exhibiting increased thermostability in its restriction endonuclease; Hendrix JD et al.; A procedure was developed for the selection of spontaneous mutants of Bacillus stearothermophilus NUB31 that are more efficient than the wild type in the restriction of phage at elevated temperatures . Inactivation studies revealed that two mutants contained a more thermostable restriction enzyme and one mutant contained three times more enzyme than the wild type . The restriction endonucleases from the wild type and one of the mutants were purified to apparent homogeneity . The mutant enzyme was more thermostable than the wild-type enzyme . The subunit molecular weight, amino acid composition, N-terminal and C-terminal amino acid residues, tryptic peptide map, and catalytic properties of the two enzymes were determined . The two enzymes have similar catalytic properties, but the molecular size of the mutant enzyme is approximately 6 to 7 kilodaltons larger than that of the wild-type enzyme . The mutant enzyme contains 54 additional amino acid residues, of which 26 to 28 are aspartate/asparagine, 8 to 15 are glutamate/glutamine, and 8 to 9 are tyrosine residues . The two enzymes contained similar amounts of the other amino acids, identical N-terminal residues, and different C-terminal residues . Tryptic peptide analyses revealed a high degree of homology between the two enzymes . The increased thermostability observed in the mutant enzyme appears to have been achieved by a mutation that resulted in the addition of amino acid residues to the wild-type enzyme . A number of mechanisms are discussed that could account for the observed difference between the mutant and wild-type enzymes.

Plasmid, 1985 May, 13(3), 211 - 4
Construction and characterization of plasmid vectors for cloning in the entomocidal organism Bacillus sphaericus 1593; Norton NB et al.; A plasmid vector for cloning in Bacillus sphaericus 1593 was constructed in B . subtilis from two parent plasmids, pBC16 and pBD64 . When characterized, the 3.9-MDa chimeric plasmid pNN101 was found to consist of the MspI fragment containing the chloramphenicol acetyltransferase (CAT) gene from pBD64 inserted into an MspI site in pBC16 . pNN101 was shown to replicate, express, and be stably maintained in B . sphaericus 1593 without affecting the mosquito larvicidal activity of this organism . A derivative of this plasmid, pNN302, was constructed in which a unique HindIII site was introduced into the CAT gene without loss of chloramphenicol resistance.

Mikrobiologiia, 1985 May-Jun, 54(3), 421 - 8
{Composition and properties of the outer cell wall layer in Bacillus brevis--the producer of gramicidin S}; Simakova IM et al.; Two membrane antigens were found by cross immunoelectrophoresis in the cell walls of Bacillus brevis var . G.-B., R form, which started to synthesize gramicidin S (20 mg per 1 ml of cultural broth) . The cell wall contained no membrane components in cells at the beginning of the logarithmic growth phase . The protein with a molecular mass of 100 kDa is a component of the cell wall outer layer . The protein is not digested by trypsin or pronase when it comprises the cell walls of cells synthesizing gramicidin S . In the preparation of isolated cell walls, this protein becomes susceptible to the action of the above proteases only when the peptidoglycan layer is broken down by lysozyme . Electron microscopy of cells treated with proteases and shadowed with a metal revealed that many cells lacked the cytoplasm . Therefore, the outer layer of B . brevis R cell wall contains small regions susceptible to the action of protease along with regions composed of the 100 kDa protein and resistant to these enzymes . It is possible that the small regions contain membrane components.

Antimicrob Agents Chemother, 1985 May, 27(5), 798 - 801
Relationship between butirosin biosynthesis and sporulation in Bacillus circulans; Nam DH et al.; The relationship between butirosin biosynthesis and certain biochemical characteristics related to sporulation in a strain of Bacillus circulans NRRL B-3313 was examined . The cellular content of dipicolinic acid increased while the amount of poly-beta-hydroxybutyrate decreased with changes in antibiotic productivity . Oligosporogenous mutants failed to synthesize the antibiotic and to degrade poly-beta-hydroxybutyrate . These observations suggest that spore formation may be related to antibiotic production in this strain of B . circulans.

Am J Vet Res, 1985 May, 46(5), 1133 - 6
DNA hybridization procedure to detect pseudorabies virus DNA in swine tissue; McFarlane RG et al.; A DNA hybridization technique was developed to detect the presence of pseudorabies virus (PRV) DNA . P Nick translated probes of high specific activity were prepared from transformed Escherichia coli plasmids into which Bacillus amyloliquefaciens H (Bam H1) restriction fragments of PRV DNA had been inserted . Swine cellular DNA and tissue culture PRV DNA were digested with Bam H1, separated by agarosegel electrophoresis, transferred onto nitrocellulose paper, hybridized to the radioactive probes, and washed under high stringency conditions; autoradiographs were then prepared . Under the optimal hybridization conditions described, the detection limit of these probes was 10(-11)g of PRV DNA . In reconstruction experiments, 3 of the selected probes cross hybridized with digested swine cellular DNA, and 4 probes did not . The addition of polyuridylic acid and polyguanylic acid to the hybridization reactions did not alter the amount of hybridization . The results indicated that this procedure may be useful for studying the latency of pseudorabies viral infection.

Biochem Biophys Res Commun, 1985 Apr 30, 128(2), 728 - 32
Germinability of coat-lacking spores of Bacillus megaterium; Nakatani Y et al.; Upon treatment with acid, the germinability of both intact and coat-lacking spores of Bacillus megaterium ATCC 19213 exhibited similar features . Namely, when the spores previously germinated by alanine in the presence of phosphate buffer were converted to H-spores by treatment with nitric acid, germination proceeded at a very low speed in a same germination medium . When H-spores converted to Ca-spores by treatment with calcium acetate and subsequently germinated, germination proceeded at a speed higher than that of native spores and occurred even in the absence of buffer . These results suggest that the site of exchangeable cations concerned with germinability must not exist in the coat.

Biochim Biophys Acta, 1985 Apr 29, 828(3), 375 - 9
Changes in the state of subunit association of lactate dehydrogenase from Bacillus stearothermophilus; Clarke AR et al.; Time-resolved measurements of the fluorescence anisotropy of an extrinsic dye-group attached to lactate dehydrogenase from B . stearothermophilus revealed that the rotational correlation time of the enzyme at low concentrations is 55 ns, while at high enzyme concentrations or in the presence of fructose 1,6-bisphosphate (Fru-1,6-P2) the correlation time increases to 95 ns . These correlation times are consistent with a change in Mr from 85 000 +/- 12 000 (dimer) to 150 000 +/- 22 000 (tetramer) and show that the tetrameric state can be induced either by raising the protein concentration or by the addition of the ligand . We have confirmed this change in molecular weight by gel-filtration experiments . In the ligand-induced tetramer, two Fru-1,6-P2 molecules are bound.

Z Gesamte Inn Med, 1985 Apr 15, 40(8), 239 - 41
{Prognosis of new cases of lung tuberculosis in 1962 and 1972 10 years later}; Werner E; At present a good prognosis of pulmonary tuberculosis is possible in the case of well-timed diagnosis and optimum treatment . Controlled use of all necessary specific drugs is a prerequisite to a success . The analogous courses of bacillary and non-bacillary pulmonary tuberculosis confirm our experiences that culture-negative respiratory tuberculosis is existing in an important number . Therefore, this factor will have to be put into our epidemiologic and clinical considerations.

Cell Immunol, 1985 Apr 15, 92(1), 1 - 13
Characterization of cytotoxic cells generated from in vitro cultures of murine bone marrow cells; Klimpel GR et al.; Bone marrow cells cultured for 5-6 days generate cytotoxic activity against a number of natural killer (NK)-susceptible tumor cells . In this study, these bone marrow cytotoxic cells were compared to cells with NK activity obtained either from spleen cells activated in vitro with interferon (IFN-alpha/beta) or mitogen or from peritoneal exudate cells (PEC) obtained 4 days after bacillus Calmette-Guerin (BCG) infection . Splenic and PEC cytotoxic cells were shown to be Thy 1.2+, NK 1.1+, Asialo GM+1, Lyt 1.2-, Lyt 2.2- . In contrast, bone marrow cytotoxic cells were Thy 1.2+, NK 1.1-, Lyt 1.2-, Lyt 2.2- and expressed low levels of Asialo GM1 antigen (Asialo GM +/- 1) . Precursor cells for bone marrow cytotoxic activity were shown to be Thy 1.2-, NK 1.1-, Lyt 1.2-, Lyt 2.2- but also expressed low levels of Asialo GM1 antigen (Asialo GM +/- 1) . Cytotoxic activity for both bone marrow and spleen cells peaked in the low-density fractions of discontinuous Percoll density gradients . The cytotoxic activity of these bone marrow cells was augmented by pretreatment with IFN (-alpha/beta, -gamma) or soluble factors (IFN free) from activated EL-4 thymoma cells . Surprisingly, the ability of bone marrow cells to generate high levels of cytotoxic activity following in vitro culture appeared to be associated primarily with mice which were of the H-2b haplotype.

Virology, 1985 Apr 15, 142(1), 98 - 111
Transcription of Bacillis subtilis plasmid pBD64 and expression of bacteriophage SPO1 genes cloned therein; Curran JF et al.; Plasmid pBD64, a vector which is useful for cloning in Bacillis subtilis (T . J . Gryczan, A . G . Shivakumar, and D . Dubnau (1980), J . Bacteriol . 141, 246-253), has at least three substantial transcription units . Two of these include the single EcoRI, XbaI, and BamHI sites, while the other includes the single BglII site . Each of these transcripts was synthesized in the counterclockwise direction, relative to the pBD64 restriction map . No transcripts were detected in the opposite direction . Infection by bacteriophage SPO1 caused a substantial decrease in each of these transcripts . No new pBD64 transcripts were detected during SPO1 infection . Various SPO1 genes, cloned at several of these pBD64 sites, were tested for expression by observing their capacity to complement SPO1 mutants . Several middle and late genes were expressed substantially, regardless of the orientation in which the fragments were inserted . Since transcription from the vector could cause expression only in one orientation, this argues that the necessary transcription originated at SPO1 promoters, and, thus, that SPO1 middle and late promoters can be active in thymine-containing DNA.

J Biochem (Tokyo), 1985 Apr, 97(4), 1085 - 92
A common linkage saccharide unit between teichoic acids and peptidoglycan in cell walls of Bacillus coagulans; Kojima N et al.; Teichoic acid-glycopeptide complexes were isolated from lysozyme digests of the cell walls of Bacillus coagulans AHU 1631, AHU 1634, and AHU 1638, and the structure of the teichoic acid moieties and their linkage regions was studied . On treatment with hydrogen fluoride, each of the complexes gave a hexosamine-containing disaccharide, which was identified to be glucosyl(beta 1----4)N-acetylglucosamine, in addition to dephosphorylated repeating units of the teichoic acids, namely, galactosyl(alpha 1----2)glycerol and either galactosyl(alpha 1----2){glucosyl(alpha 1----1/3)}glycerol (AHU 1638) or galactosyl(alpha 1----2){glucosyl(beta 1----1/3)}glycerol (AHU 1631 and AHU 1634) . From the results of Smith degradation, methylation analysis, and partial acid hydrolysis, the teichoic acids from these strains seem to have the same backbone chains composed of galactosyl(alpha 1----2)glycerol phosphate units joined by phosphodiester bonds at C-6 of the galactose residues . The presence of the disaccharide, glucosyl(beta 1----4)N-acetylglucosamine, in the linkage regions between teichoic acids and peptidoglycan was confirmed by the isolation of a disaccharide-linked glycopeptide fragment from each complex after treatment with mild alkali and of a teichoic acid-linked saccharide from each cell wall preparation after treatment with mild acid . Thus, it is concluded that despite structural differences in the glycosidic branches, the teichoic acids in the cell walls of the three strains are linked to peptidoglycan through a common linkage saccharide, glucosyl (beta 1----4) N-acetylglucosamine.

Biochem J, 1985 Apr 1, 227(1), 183 - 9
Electrophoretic characterization of hepatic alkaline phosphatase released by phosphatidylinositol-specific phospholipase C . A comparison with liver membrane and serum-soluble forms; Kominami T et al.; Alkaline phosphatase was solubilized from plasma membrane of rat liver with butanol-ol, bile acids or sodium deoxycholate, and electrophoretically compared with a soluble form in serum which was derived from the liver . The three enzyme preparations from the plasma membrane migrated at the same position on polyacrylamide-gel electrophoresis in the presence of either Triton X-100 or sodium dodecyl sulphate . The mobility of them, however, was distinctly different from that of the serum-soluble form of the liver-derived alkaline phosphatase . On the other hand, phosphatidylinositol-specific phospholipase C isolated from Bacillus cereus was used to release alkaline phosphatase from plasma membrane . The released alkaline phosphatase was demonstrated to have the same mobility as the serum-soluble form on polyacrylamide-gel electrophoresis in the presence or absence of detergents . The phospholipase C also converted the butan-1-ol-extracted membrane form into the serum-soluble form . The results suggest that release of alkaline phosphatase from the liver into serum is not simply caused by a detergent effect of bile salts, but involves an enzymic hydrolysis of phosphatidylinositol, with which alkaline phosphatase may strongly interact in the membrane.

J Urol, 1985 Apr, 133(4), 598 - 601
The management of superficial bladder tumors and carcinoma in situ with intravesical bacillus Calmette-Guerin; deKernion JB et al.; A phase II study was performed to assess the role of bacillus Calmette-Guerin as a prophylaxis against recurrent stages O and A bladder tumors, and in the treatment of existing superficial bladder tumors and carcinoma in situ . Tice strain bacillus Calmette-Guerin (1 vial, 2 to 8 times 10(8) organisms in 60 cc saline) was instilled intravesically without cutaneous inoculation . Instillations were given weekly for 6 weeks and then monthly or until recurrence in 22 patients with a history of recurrent tumors, while 22 with existing stages O and A transitional cell carcinoma, and 19 with carcinoma in situ were treated weekly for 8 weeks and then monthly for 12 months or until failure . Complications included cystitis in 88 per cent of the patients (severe in 20 per cent), fever in 15 per cent, a flu-like syndrome in 13 per cent, edema and pruritis in 1.5 per cent, and ureteral stenosis in 1.5 per cent . Twelve patients (19 per cent) did not complete the study owing to toxicity . Of the patients in the prophylaxis group 67 per cent have had no tumor recurrence 10 to 26 months (mean 15 months) after therapy . Of the patients with existing tumors 36 per cent had complete regression following bacillus Calmette-Guerin therapy and 23 per cent had a partial response . Among the patients with carcinoma in situ 13 (68 per cent) had reversal to normal urothelium and 3 (16 per cent) had marked improvement . None of the patients had recurrence at 11 to 20 months . Intravesical Tice strain bacillus Calmette-Guerin is effective as a prophylaxis against recurrent superficial bladder tumors and in the treatment of carcinoma in situ.

Appl Environ Microbiol, 1985 Apr, 49(4), 904 - 7
Mechanism of action of aflatoxin B1 in Bacillus megaterium; Tiwari RP et al.; Bacillus megaterium cells from various growth phases were equally susceptible to the lethal effects of aflatoxin B1 . Known surfactants (EDTA and Tween-80) accentuated the effects of aflatoxin B1 . Viability and inulin uptake in aflatoxin B1-exposed cells decreased considerably . The effect was concentration dependent . A straight-line relationship observed in the death curve indicated a single target for aflatoxin B1 action in B . megaterium . Leakage of intracellular constituents in B . megaterium was also concentration dependent, and this can be related to the extent of cell membrane damage.

Indian J Lepr, 1985 Apr-Jun, 57(2), 278 - 81
Immunological potential of a cultivable mycobacterial strain M . habana against leprosy bacillus in mouse foot pad; Singh NB et al.; A strain of a typical mycobacteria M . habana originally afforded protection against M . tuberculosis challenge in mice, was tested for its immunological potential against leprosy bacillus in the mouse foot pad . The vaccine strain M . habana has arrested the growth of M . leprae into the mouse foot pad better than BCG (Phipps) and unvaccinated control.

J Appl Bacteriol, 1985 Apr, 58(4), 351 - 4
A note on an antimicrobial agent in disc paper and a modified assay procedure for detecting antimicrobials in milk; Goodman GD et al.; Commercially available disc paper used for antibiotic sensitivity tests was shown to contain an unidentified, water-soluble agent inhibitory to Bacillus stearothermophilus which gave false positive results in an antimicrobial assay for milk . The agent however, was found to be inactive against a range of common pathogenic bacteria . It was easily removed by washing the discs in distilled water before use.

Mol Gen Mikrobiol Virusol, 1985 Apr, (4), 22 - 5
{Effect of the pC194 plasmid on the development of Bacillus thuringiensis phages}; Lugovskaia EV et al.; Bacillus thuringiensis var . galleriae strains were transformed by plasmid pC194, coding for chloramphenicol resistance (CmR) . Efficiency of plating and the yields of bacteriophages Tg13 and Tg27 maturating in CmR transformant cells were decreased for 2-3 orders as compared with the ones in parental strains . The CmR transformants are characterized by the increased level of spontaneous induction of bacteriophage Tg22.

J Biochem (Tokyo), 1985 Apr, 97(4), 977 - 82
Preparation of non-reducing-end substituted p-nitrophenyl alpha-maltopentaoside (FG5P) as a substrate for a coupled enzymatic assay for alpha-amylases; Omichi K et al.; p-Nitrophenyl O-6-deoxy-6-{(2-pyridyl)amino}-alpha-D-glucopyranosyl-(1----4)-O-alpha-D - glucopyranosyl-(1----4)-O-alpha-D-glucopyranosyl-(1----4)-O-alpha-D- glucopyranosyl-(1----4)-alpha-D-glucopyranoside, FG5P, was prepared, taking advantage of the action of Bacillus macerans cyclodextrin glucanotransferase on a mixture of O-6-deoxy-6-{(2-pyridyl)-amino}-alpha-D-glucopyranosyl-(1----4)-O-alpha- D- glucopyranosyl-(1----4)-O-alpha-D-glucopyranosyl-(1----4)-O-alpha-D- glucopyranosyl-(1----4)-O-alpha-D-glucopyranosyl-(1----4)-D-glucose and p-nitrophenyl alpha-glucoside . The maltopentaose derivative is resistant to alpha-glucosidase and is suitable as a substrate for the alpha-amylase assay coupled with alpha-glucosidase in which the activity of alpha-amylase is determined by measuring the amount of p-nitrophenol liberated by alpha-glucosidase from p-nitrophenyl alpha-glucoside and p-nitrophenyl alpha-maltoside produced by the action of alpha-amylase . This alpha-amylase assay method was applied for determination of alpha-amylases in human serum.

Antibiot Med Biotekhnol, 1985 Apr, 30(4), 288 - 92
{Differences in the cytotoxicity mechanisms of pancreatic and microbial ribonucleases}; Kurinenko BM et al.; The effect of pancreatic and microbial ribonucleases (RNAses) on incorporation of labeled precursors, i . e . 14C-uridine, 14C-thymidine and 14C-glycine into the biopolymers of transplantable cell cultures, as well as their effect on radioactivity of the acid insoluble fraction of the cells labeled with 14C-orotic acid was studied . It was shown that the pancreatic RNAses and RNAses of Streptomyces rimosus in a dose of 1 ED50 determined by the index of the cell monolayer intactness inhibited incorporation of the labeled precursors . The inhibition was rather deep and almost equal . The enzymes lowered also radioactivity of the cells labeled with 14C-orotic acid in proportion to the time of the cell incubation in the presence of the enzymes . On the contrary, RNAses of Bacillus sp . used in analogous doses was significantly less intensive in inhibition of the precursor incorporation and almost did not lower radioactivity of the acid insoluble fraction of the cells totally labeled with 14C-orotic acid.

Antibiot Med Biotekhnol, 1985 Apr, 30(4), 257 - 61
{Superoxide dismutase activity of Bacillus brevis var . G.-B . cells, producer of gramicidin C, in relation to cultivation conditions}; Udalov TP et al.; The activity of superoxide dismutase (SOD) in the cells of the microorganisms producing gramicidin S, B . brevis var . G.-B., ranged from 7 to 22 units per mg protein . When the culture was grown in a fermenter, the decrease in the concentration of dissolved oxygen in the medium to the minimum values corresponded to the decrease in the enzyme activity . By the end of the fermentation process, when consumption of oxygen by the culture lowered and its level in the medium increased, the enzyme activity increased . When the culture was grown in flasks, no effect of the medium aeration level on the enzyme activity was noted . This might be associated with rather high levels of dissolved oxygen in the medium in the flasks under all aeration conditions . Addition of manganese ions to the cultivation medium activated SOD in the cells of B . brevis . Synthesis of gramicidin S by the bacterial culture had no effect on the activity of SOD in the cells.

FEBS Lett, 1985 Mar 25, 182(2), 265 - 8
Relationship between Na+-dependent cytoplasmic pH homeostasis and Na+-dependent flagellar rotation and amino acid transport in alkalophilic Bacillus; Sugiyama S et al.; The cytoplasmic pH homeostasis of alkalophilic Bacillus strains required the presence of Na+ in the medium, and Li+ was found to be equivalently substitutable for Na+ . Flagellar rotation and amino acid transport of these bacteria also required Na+ but Li+ was not substitutable for Na+ . Na+ concentration of about 1 mM was enough for the cytoplasmic pH homeostasis, while more than 10 mM Na+ was required for the full activities of flagellar rotation and amino acid transport . The addition of 150 mM ethanolamine to the cells at pH 9.6 disrupted the pH homeostasis and increased the cytoplasmic pH close to the external pH . Under this condition, however, flagellar rotation and amino acid transport were not so much affected . Thus, it is clear that flagellar rotation and amino acid transport themselves require the presence of Na+ in the medium, independent of the Na+-dependent cytoplasmic pH homeostasis.

Cancer, 1985 Mar 15, 55(6), 1236 - 43
Prospectively randomized trial of adjuvant active-specific immunotherapy for human colorectal cancer; Hoover HC Jr et al.; Over the last four years a guinea pig model of active-specific immunotherapy (ASI) with a syngeneic tumor cell:bacillus calmette-Guerin (BCG) vaccine was translated into a prospectively randomized, controlled clinical trial in patients with colorectal cancer . Primary tumors from patients undergoing standard surgical resection were dissociated enzymatically and cryopreserved by techniques that maintain cell viability . Patients with transmural extension of tumor or nodal metastases were randomized into groups treated by resection alone (control) or resection plus ASI . With a mean follow-up of 28 months (range, 14-24), only 3 of 20 treatment patients had recurrences and none have died, whereas 9 of 20 control patients had recurrences and 4 died . These differences are statistically significant and are sufficiently encouraging to warrant expansion of these studies into other research centers.

Biochemistry, 1985 Mar 12, 24(6), 1298 - 306
Methyl branching in short-chain lecithins: are both chains important for effective phospholipase A2 activity?
DeBose CD, Burns RA Jr, Donovan JM, Roberts MF.
Several seven-carbon fatty acyl lecithins with varied acyl chain branching have been synthesized and characterized as potential phospholipase A2 substrates . Micellar bis(4,4-dimethylpentanoyl) phosphatidylcholine, bis(5-methylhexanoyl)phosphatidylcholine, bis(3-methylhexanoyl)phosphatidylcholine, and bis(2-methylhexanoyl)phosphatidylcholine are poor substrates for phospholipase A2 (Naja naja naja) . These branched lecithins also inhibit the hydrolysis of diheptanoylphosphatidylcholine by the enzyme with Ki values comparable to or smaller than the apparent Km of the linear compound . The terminally branched lecithins are excellent substrates for another surface-active hydrolytic enzyme, phospholipase C from Bacillus cereus . When only one acyl chain bears a methyl group, the hybrid lecithins 1-heptanoyl-2-(2-methylhexanoyl)phosphatidylcholine and 1-(3-methylhexanoyl)-2-heptanoylphosphatidylcholine are substrates comparable to diheptanoylphosphatidylcholine . Analysis of micellar structure and dynamics by 1H and 13C NMR spectroscopy, quasi-elastic light scattering, and comparison of critical micellar concentrations indicates little significant difference in the conformation and dynamics of these seven-carbon fatty acyl lecithin micelles, even when the methyl groups are adjacent to the carbonyls . Phospholipase A2 UV difference spectra induced by phospholipid binding imply different enzyme conformations or aggregation states caused by linear-chain and asymmetric-chain lipids compared to bis(methylhexanoyl)phosphatidylcholines . The differences in hydrolytic activity of phospholipase A2 against the branched-chain micellar lecithins can then be attributed to an enzyme-lipid interaction at the active site . The species with both fatty acyl chains branched bind to phospholipase A2 but are not turned over rapidly . Since poor enzymatic activity only occurs for lecithins with both chains methylated, the interaction of both chains with the enzyme must be important for catalytic efficiency.

Biochim Biophys Acta, 1985 Mar 8, 838(3), 302 - 11
Purification and characterization of pentobarbital-induced cytochrome P-450BM-1 from Bacillus megaterium ATCC 14581; Schwalb H et al.; When Bacillus megaterium ATCC 14581 is grown in the presence of barbiturates, a cytochrome P-450-dependent fatty acid monooxygenase (Mr 120000) is induced (Kim, B.-H . and Fulco, A.J . (1983) Biochem . Biophys . Res . Commun . 116, 843-850) . Gel filtration chromatography of a crude monooxygenase preparation from pentobarbital-induced B . megaterium indicated that not all of the induced cytochrome P-450 present in the extract was accounted for by this high-molecular-weight component . Further purification revealed the presence of two additional but smaller cytochrome P-450 species . The minor component, designated cytochrome P-450BM-2, had a molecular mass of about 46 kDa, but has not yet been completely purified or further characterized . The major component, designated cytochrome P-450BM-1, was obtained in pure form, exhibited fatty acid monooxygenase activity in the presence of iodosylbenzenediacetate, and has been extensively characterized . Its Mr of 38000 makes it the smallest cytochrome P-450 yet purified to homogeneity . Although it is a soluble protein, a complete amino acid analysis indicated that it contains 42% hydrophobic residues . By the dansyl chloride procedure the NH2-terminal amino acid is proline; the penultimate NH2-terminal residue is alanine . The absolute absorption spectra of cytochrome P-450BM-1 show maxima in the same general regions as do P-450 cytochromes from mammalian or other bacterial sources, but they differ in detail . The oxidized form of P-450BM-1 has absorption maxima at 414, 533 and 567 nm, while the reduced form has peaks at 410 and 540 nm . The absorption maxima for the CO-reduced form of P-450BM-1 are found at 415, 448 and 550 nm . Antisera from rabbits immunized with pure P-450BM-1 strongly reacted with and precipitated this P-450, but showed no detectable affinity for either the 46 kDa P-450 or the 120 kDa fatty acid monooxygenase.

Southeast Asian J Trop Med Public Health, 1985 Mar, 16(1), 54 - 8
Bioassays of Bacillus sphaericus (strain 1593) against mosquitoes of public health importance in Malaysia; Cheong WC et al.; The pathogenicity of Bacillus sphaericus strain 1593 was tested against laboratory-reared larvae of four local species of mosquitoes of public health importance in Malaysia; Aedes aegypti, Anopheles balabacensis, Mansonia uniformis and Culex quinquefasciatus . The bacteria was shake-cultured at 28 +/- 1 degrees C for three days, using Glucose-Yeast Extract Salts medium . After which, the spores and vegetative cells were harvested and stored at 4 degrees C before use . Conditions for bioassays were mean temperature of 25 +/- 1 degrees C and relative humidity 65 +/- 5.0 . Twenty third-instar larvae of each species were assayed in 90 ml of diluted spore solution . Each concentration and a control were replicated three times for each bioassay . Larval mortalities at 24 hours and 48 hours were taken and analyzed through Probit Analysis using a computer (IBM 370) . LC50 values after 48 hours of exposure showed an increasing order of larval susceptibility as follows: Ae . aegypti (417.70 x 10(4)), An . balabacensis (45.84 x 10(4)), Ma . uniformis (18.23 x 10(4)) and Cx . quinquefasciatus (4.14 x 10(4) spores/ml) . With the ability to kill 90% of the Cx . quinquefasciatus larvae tested with just a concentration of 10(5) spores/ml, B . sphaericus (strain 1593) has shown good potential as a biocontrol agent for this species of mosquito.

Rev Infect Dis, 1985 Mar-Apr, 7 Suppl 1, S185 - 90
Selective immunization: protection of the individual; Dudgeon JA; Selective immunization was adopted in the United Kingdom on account of doubts about the duration of vaccine-induced immunity . Selective immunization of a particular age group had already been established as an effective procedure with bacille Calmette-Guerin (BCG) vaccine, so that practical experience was available . Initially, girls aged 11-14 years were vaccinated, but the program was extended over a 14-year period to include other groups, mainly susceptible women and other at-risk individuals . Cases of congenital rubella continue to occur, although there are signs of a decline in incidence . The number of terminations of pregnancy related to rubella in any form is high . The vaccination acceptance rate for girls aged 10-14 years increased from 35% in 1972 to 85% in 1982 . The low rate of measles vaccination is considered to be a major reason for continuing with the present scheme rather than changing to the American or Swedish schemes of vaccination of preschool children with combined vaccines . A new initiative to promote rubella vaccination has recently been introduced in the United Kingdom in which charitable institutions would work together with health authorities to promote immunization against rubella.

J Epidemiol Community Health, 1985 Mar, 39(1), 20 - 6
Tuberculosis: spatial and demographic incidence in Bradford, 1980-2; Froggatt K; Between 1980 and 1982 the incidence of tuberculosis in Bradford Health Authority was approximately 20 times higher among the New Commonwealth and Pakistani population than the rest of the population . It was also possible to see a clustering of cases within this time period, spatially and demographically--in certain age groups and by sex . The difference between the two populations was not due to race but rather reflects the socioeconomic position that the New Commonwealth and Pakistani population has within Bradford as a whole . The incidence of non-Asian tuberculosis rose in 1982 due to an outbreak among unvaccinated young adults . Much still needs to be done to eradicate the environmental conditions within which the tubercle bacillus thrives as well as to educate the population at risk.

Fam Pract, 1985 Mar, 2(1), 30 - 4
Family tuberculosis contacts: resource-contingent management; Pust RE; Recent findings in tuberculosis research have questioned the efficacy of bacille Calmette-Guerin (BCG) vaccination and demonstrated the effectiveness of combined-drug chemotherapy and isoniazid (INH) chemoprophylaxis, both in regimens of under 12 months duration . Because of the renewed emphasis on drug treatment in tuberculosis control, family physicians and the health personnel they supervise need to be involved in this effort . Despite differences in health care resources in different regions, rational and effective management of active cases and their contacts in the family can be devised . While the priority remains treatment of the active index case, family physicians have a unique opportunity to utilize family relationships to find and to treat other active cases and to reinforce compliance with INH chemoprophylaxis by high-risk family contacts.

Virology, 1985 Mar, 141(2), 190 - 200
Structure of phage phi 29 connector protein assembled in vivo; Carrascosa JL et al.; The protein p10 that forms the connector of phage phi 29, has been produced in Escherichia coli harboring a plasmid that carried the gene coding for this protein . The connector protein is assembled in a 13.4-S oligomer that has an apparent molecular weight of 460,000, suggesting that it is a dodecamer . The purified oligomers have been studied by electron microscopy of the isolated particles as well as by image-processing techniques (Fourier and rotational filtering) of artificially induced two-dimensional aggregates . The results show that the purified p10 is assembled in a circular structure with a hole in its center and 12 morphological units in the periphery . Both the morphology and the dimensions of this p10 oligomer are very similar to those of the upper neck collar extracted from phi 29 viral particles . The results strongly suggest the close relationship between the p10 oligomers assembled in E . coli and the ones produced in phi 29 infected Bacillis subtilis.

Arch Microbiol, 1985 Mar, 141(2), 143 - 50
Regulation of lysine and dipicolinic acid biosynthesis in Bacillus brevis ATCC 10068: significance of derepression of the enzymes during the change from vegetative growth to sporulation; Rao AS; Lysine biosynthetic pathway enzymes of Bacillus brevis ATCC 1068 were studied as a function of stage of development (growth and sporulation) . The synthesis of aspartic-2-semialdehyde dehydrogenase (ASA-dehydrogenase), dihydrodipicolinate synthase (DHDPA-synthase), DHDPA-reductase and diaminopimelate decarboxylase (DAP-decarboxylase) was found not to be co-regulated, since lysine was not a co-repressor for these enzymes . Unlike the aspartokinase isoenzymes, the other enzymes of the lysine pathway were not derepressed in thiosine-resistant, lysine-excreting mutants . Thus, the aspartokinase isoenzymes were the key enzymes during growth and regulation of lysine biosynthesis through restriction of L-ASA synthesis via feedback control by lysine on the aspartokinases was therefore suggested . In contrast to other Bacillus species, the levels of the lysine biosynthetic pathway enzymes of strain ATCC 10068 were not derepressed during the change from vegetative growth to sporulation . Two control mechanisms, enabling the observed preferential channelling of carbon for the synthesis of spore-specific diaminopimelic acid (DAP) and dipicolinic acid (DPA) were a) loss of DAP-decarboxylase, b) inhibition of DHDPA-reductase by DPA . Increase in the level of the DAP pool during sporulation, as a consequence of the loss of DAP-decarboxylase, and its relevance to the non-enzymatic formation of DPA has been discussed.

Prikl Biokhim Mikrobiol, 1985 Mar-Apr, 21(2), 184 - 9
{Properties of the phospholipases C from Bacillus cereus}; Gerasimene GB et al.; Three different phospholipases C--the so-called phospholipase C which hydrolyses phosphatidylcholine (Pch-PLC), sphingomyelinase (SM-PLC) and phosphatidylinositol hydrolysing phospholipase C (PI-PLC)--were separated from the culture filtrate of Bacillus cereus using column chromatography on DEAE-sephadex A-50 . The pI values were estimated to be 5.0 +/- 0.3 for PI-PLC and 5.3 +/- 0.2 for SM-PLC . The effect of some bivalent cations was studied . Metal ions had no effect on the activity of PI-PLC, whereas Mg2+ and Co2+ in concentrations from 1 to 5 mM highly activated SM-PLC . Mg2+ failed to activate PCh-PLC, and Co2+ even inhibited it . EDTA of o-phenantroline had a rather inhibitory effect on Pch-PLC, but they almost did not affect SM-PLC.

Am Rev Respir Dis, 1985 Mar, 131(3), 466 - 9
Life-threatening complications associated with Bacillus cereus pneumonia; Bekemeyer WB et al.; Bacillus species are identified as pathogens in lung and pleural space infections with increasing frequency . We report a patient who developed life-threatening complications of Bacillus cereus pneumonia, including massive hemoptysis, acute respiratory failure, tension pneumothorax, empyema, and bronchopleural fistula . We also review the pertinent literature concerning the associated underlying disorders, complications, and therapy of Bacillus species pulmonary infections.

Mikrobiologiia, 1985 Mar-Apr, 54(2), 181 - 5
{Amino acid and mineral element content and the activity of various enzymes in germinating spores of Bacillus thuringiensis}; Alekseev AN et al.; Changes in the content of dipicolinic acid and mineral elements were studied in the process of Bacillus thuringiensis spore germination . The spores released up to 28% of dipicolinic acid and 18% of calcium at the activation stage, and 93 and 91%, respectively, at the initiation stage . At the same time, the content of Mg, Mn, Zn and P decreased while K, Na and Fe accumulated in the spores . The activities of total and serine proteases, alkaline phosphatase, NADH dehydrogenase and aldolase increased in the extract of initiated spores . The content of glutamate decreased in the free amino acid pool as early as by the 30th second of the initiation stage.

Mol Biol (Mosk), 1985 Mar-Apr, 19(2), 349 - 58
{Immune electron microscopy in the study of biological matter}; Kretova AF et al.; A brief review of literature data and our investigations on the antibodies used for specific labeling in electron microscopy is presented . Considered are the problems connected with structure and function of separate components of bacterial viruses revealed by means of specific antibodies . The results of fine differentiation of antigenic components in the case of phages of the colidysentery group allowed to elucidate the functional role of the adsorption apparatus in the course of phage interaction with the bacterial cell . The topology of structural proteins (gene-products 35, 36, 37) of the tail's long strands for phages T4, DDVI+h and DDVIh is determined . Antigenic properties of proteins that are found in the composition of two forms of Bacillus mycoides are demonstrated immune-electronmicroscopically . On the basis of this finding, a conclusion was made that one of these phages acted as precursor, the other--as satellite during the simultaneous development of these phages in the bacterial cell . It was also established that temperate and virulent phages are related antigenically, which proves that lysogenic bacteria can be one of the phage sources on the environmental conditions . Visualization of non-ribosomal genes of procaryots that code for structural proteins of a defective phage proves the efficiency of the immune-electronmicroscopic method for investigating of biological objects.

Clin Allergy, 1985 Mar, 15(2), 101 - 15
A longitudinal study of possible allergy to enzyme detergents; Pepys J et al.; One hundred and thirty-six subjects with asthma and/or hay fever were surveyed for possible sensitization to the Bacillus licheniformis protease present in the commercial antigenically identical enzymes (Alcalase and Maxatase) used in household detergents . Prick-test reactions reported as minimally positive were given by one subject to both commercial enzyme preparations at the same standard non-irritating concentrations and by four others to one but not the other antigenically identical enzyme . Two of the five, and an additional seven, gave weak prick tests to the purified enzyme (Koch-Light Protease) at test protein concentrations 100 times stronger and to which non-specific irritation occurs . The findings are strongly against interpretation of these reactions as positive and were on no occasion comparable with those in sensitive workers . The skin-test reactions are attributable to non-specific effects . Neither enzyme-specific RAST IgE tests nor clinical histories suggested clinical sensitivity . Enzyme-specific RAST IgE for eighty-eight subjects over a period of 5 years showed no correlation with exposure . None had developed specific IgE levels like those of sensitized workers, and their RAST levels did not differ significantly from cord blood, unexposed subjects and prick-test-negative workers . The eight with the largest increase in specific IgE were prick-test-negative and their clinical reactions to enzyme detergents were comparable to those of non-enzyme detergents . There is no evidence from this study that exposure to modern, non-dusty household enzyme-containing detergents is likely to lead to clinical sensitization.

Appl Environ Microbiol, 1985 Mar, 49(3), 706 - 8
Only part of the protoxin gene of Bacillus thuringiensis subsp . berliner 1715 is necessary for insecticidal activity; Wabiko H et al.; Escherichia coli strains harboring deletion mutations of the insecticidal protoxin gene of Bacillus thuringiensis subsp . berliner 1715 were constructed . Although these strains did not produce intact protoxin, cell extracts from one of the mutants were extremely toxic to tobacco hornworm (Manduca sexta) larvae, indicating that only a part of the protoxin gene is required for insecticidal activity.

J Am Mosq Control Assoc, 1985 Mar, 1(1), 69 - 75
Control of snow pool mosquitoes with Bacillus thuringiensis serotype H-14 in mountain environments in California and Oregon; Eldridge BF et al.; Studies were conducted in mountainous areas of California and Oregon to test the effectiveness of Bacillus thuringiensis serotype H-14 in controlling larvae of snow pool Aedes mosquitoes, and also the effect of such larval control on the density of adult mosquitoes . California and Oregon studies showed that a wettable powder formulation of B.t.i . was effective in controlling mosquito larvae . In Oregon, treatment was effective even at water temperatures as low as 5 degrees C . Sampling of adult mosquitoes at campgrounds where intensive larval control was done failed to demonstrate a lowering of adult mosquito density in comparison with untreated control areas . Flight range studies showed that snow pool Aedes species can fly distances of up to 2 km, suggesting that infiltration from outside the treated area was one cause of failure . Inadequate seasonal timing appeared to be an additional factor . Using a simple computer model, we estimated that larval mosquito control with B.t.i . could be done for about $5,000 per season for a 500 ha area similar to our test area, a cost of $10 per ha ($4 per acre).

J Am Mosq Control Assoc, 1985 Mar, 1(1), 63 - 8
A chironomid (Diptera: Chironomidae) midge population study and laboratory evaluation of larvicides against midges inhabiting the lagoon of Venice, Italy; Ali A et al.; Chironomid larval densities in the saltwater lagoon of Venice, Italy, were assessed in the spring of 1984 . Four organophosphates; chlorpyrifos, temephos, fenthion and fenitrothion, and three pyrethroids; cypermethrin, permethrin and deltamethrin, were tested in the laboratory against field-collected larvae . Three industrial formulations of Bacillus thuringiensis var . israelensis (B.t.i.) were also tested as midge larvicides . Only Chironomus salinarius occurred in the benthic samples taken from different sections of the lagoon . The densities of this species ranged from 0 to 38,976 larvae/m2 . The highest mean larval density of 15,673/m2 was encountered in a section of the lagoon adjacent to Venice airport and receiving large quantities of raw sewage . The lowest mean density (less than 1 larva/m2) existed in another area of the lagoon receiving discharge from chemical industry . Cypermethrin and permethrin were 21-233X more active against the larvae than the four organophosphates . Chlorpyrifos was the most active organophosphate . Formulations of B.t.i . were economically ineffective against the larvae.

J Am Mosq Control Assoc, 1985 Mar, 1(1), 51 - 5
Efficacy of Bacillus thuringiensis (H-14) for larval Aedes mosquito control in intermountain meadows in Wyoming; Jones CJ et al.; One square meter field enclosures made of steel flashing, and natural ponds were used to determine the efficacy of Bactimos formulations of Bacillus thuringiensis (H-14) for control of late instar Aedes dorsalis, Ae . fitchii and Ae . melanimon mosquito larvae in intermountain meadows . Low temperatures caused decreased efficacy of the formulations tested in the laboratory, and spring temperature extremes lowered efficacy in field tests . Adequate control of fourth instar larvae was obtained in field studies in a 24 hour period when water temperatures were greater than or equal to 12 degrees C at a treatment rate of 0.1 mg/liter.

J Am Mosq Control Assoc, 1985 Mar, 1(1), 38 - 42
Efficacy of granular formulations of Bacillus thuringiensis (H-14) for the control of Anopheles larvae in rice fields; Lacey LA et al.; Three granular formulations of Bacillus thuringiensis (H-14) were applied to dense stands of maturing and mature rice for control of Anopheles crucians and An . quadrimaculatus . Aerial applications of the Vectobac granule (200 ITU/mg) at 5.6, 11.2 and 22.4 kg/ha to 0.4 ha plots resulted in 92, 94 and 96% reduction 48 hr after application, respectively, in populations predominantly consisting of late instars . The Bactimos granule (175 ITU/mg) was applied by aircraft to 0.4 ha plots and by a Cyclone seeder to 100 m2 plots at 2.8, 5.6 and 11.2 kg/ha, resulting in 100% reduction after 48 hr in natural populations of predominantly early instar larvae at all 3 rates with each type of application . Older instars confined to sentinel cages responded with 92, 98 and 100% mortality, respectively . Complete mortality was also observed at the same time in natural populations of predominantly young larvae in similar plots treated with Teknar granules (104 ITU/mg) at 1.7, 3.0 and 7.5 kg/ha . Near complete mortality (98-99%) was also observed in the older larvae used in the sentinel cages.

Med Clin North Am, 1985 Mar, 69(2), 243 - 56
Gram-negative bacillary meningitis; LeFrock JL et al.; The incidence of gram-negative bacillary meningitis has increased significantly in the past two decades . Approximately two thirds of all reported cases have occurred after neurosurgical procedures . With the development of the newer cephalosporins, the overall mortality rate has decreased from 40 to 80 per cent to 10 to 20 per cent.

J Biochem (Tokyo), 1985 Mar, 97(3), 911 - 21
Ectoenzyme release from rat liver and kidney by phosphatidylinositol-specific phospholipase C; Taguchi R et al.; Ectoenzyme release from rat liver and kidney by phosphatidylinositol (PI)-specific phospholipase C of Bacillus thuringiensis was studied . Alkaline phosphatase and 5'-nucleotidase were released from rat kidney slices to extents of up to 60% and 30%, respectively . Release of alkaline phosphatase was observed at lower amounts of PI-specific phospholipase C than that of 5'-nucleotidase . Both enzymes were more easily released from microsomal fractions or free cells . From kidney cells, alkaline phosphatase was released without cell lysis, and more than 80% release of alkaline phosphatase was observed at 3.8% hydrolysis of PI . Isoelectric focusing profiles of alkaline phosphatase released by PI-specific phospholipase C were significantly different from the control in the cases of both rat liver and kidney . Lubrol-solubilized alkaline phosphatase was eluted at the void volume of a Toyopearl HW-55 column, while the enzyme obtained by further treatment with PI-specific phospholipase C was eluted in the lower-molecular-weight region corresponding to 100,000-110,000 daltons . Furthermore, Lubrol-solubilized phosphatase became more thermostable on treatment with PI-specific phospholipase C.

Acta Radiol Oncol, 1985 Mar-Apr, 24(2), 189 - 91
Influence of catalase on the radiation sensitizing effect of misonidazole; Gazso GL et al.; The radiation modifying action of misonidazole and catalase was investigated in Bacillus megaterium spores at various oxygen concentrations . Catalase (120 micrograms/ml) decreased the radiation sensitizing action of misonidazole . Misonidazole as an electron affinic radiation sensitizer enhanced the build up of H2O2, thus promoting the reaction with catalase . Protection by catalase was not enough to eliminate the total radiation sensitizing effect of misonidazole.

Cancer Treat Rep, 1985 Mar, 69(3), 251 - 8
Combined-modality treatment of inoperable lung cancer (i.v . immunotherapy, chemotherapy, and radiotherapy); Robinson E et al.; Patients with inoperable non-small cell carcinoma of the bronchus were treated by iv methanol extraction residue of bacillus Calmette-Guerin (MER) followed by combination chemotherapy with methotrexate, doxorubicin, cyclophosphamide, and lomustine (MACC) . Radiotherapy was added in patients with localized disease . MER was given iv in four weekly courses . The dose ranged between 0.1 and 0.8 mg/m2/course and was determined according to the skin reactivity to MER . None of the 19 patients treated with iv MER had an objective tumor response during immunotherapy . Five of 16 evaluable patients receiving MACC (31%) achieved partial response . The overall median survival for all 17 patients treated was 11 months according to the protocol . During immunotherapy the following side effects were observed: all patients had fever and chills and most had malaise, nausea and/or vomiting, and headache . Transient abnormalities of liver function tests were found in six patients, and in one patient roentgenographic changes suggestive of lung granulomas were seen . The side effects were more severe during the first course than in the further treatments . Side effects of the MACC combination were comparable to those observed in previously reported studies . Immunological monitoring performed during immunotherapy revealed that iv MER had an unfavorable effect on the immune system . Following four courses there was a decrease in the in vivo skin reactivity to MER and to five recall antigens . Twenty-four hours after the initiation of iv MER there was a tendency for decrease in the lymphoproliferation to phytohemagglutinin, an increase in the indices of the adherent cell suppressor system and the prostaglandin-related suppressor system, and an increase in the percentage of adherent mononuclear cells . No consistent changes in serum lysozyme were found . A transient increase in the total number of peripheral blood leukocytes and a decrease in the number of lymphocytes were noticed 24 hours after iv MER . Taking into account the results of the immunological studies and the overall therapeutic results, we do not feel justified in continuing to use this combined modality treatment.

J Am Mosq Control Assoc, 1985 Mar, 1(1), 1 - 7
The story of Bacillus thuringiensis var . israelensis (B.t.i.); Margalit J et al.; The isolation and the characteristics of Bacillus thuringiensis var . israelensis is reviewed . Included in the review are full descriptions of the nomenclature, mosquito target range of this potent mosquitocidal bacterium, as well as the genetics and biochemistry of the toxins.

J Cell Sci, 1985 Mar, 74, 137 - 52
Electron probe X-ray microanalysis of the effects of Bacillus thuringiensis var kurstaki crystal protein insecticide on ions in an electrogenic K+-transporting epithelium of the larval midgut in the lepidopteran, Manduca sexta, in vitro; Gupta BL et al.; An alkaline hydrolysate of Bacillus thuringiensis var kurstaki HD1 (Btk) parasporal crystals was administered at 25 micrograms ml-1 (f.c.) to isolated, short-circuited, midguts of tobacco hornworm (Manduca sexta) larvae . The short-circuit current (s.c.c.), a precise measure of K+ active transport, was inhibited by 78% in 10 min in Btk-treated midguts as compared to controls . The elemental concentrations of K, together with Na, Mg, P, S, Cl and Ca, as well as the water content, were determined by electron probe X-ray microanalysis (EPXMA) in the muscle cells, columnar cells and goblet cells, as well as in the extracellular goblet cavity and the bathing media . The average K concentration in the goblet cell cavity was 129 mmol/kg wet wt in control midguts but only 37 mmol/kg wet wt in Btk-treated midguts . The elemental concentrations, including that of K, in other cell compartments were much less affected by Btk, but a rise in total cell calcium is suggested . It has been previously suggested that in vitro Btk acts specifically on limited regions of the apical membrane of the midgut epithelial cells . The simplest interpretation of the EPXMA results would be that initially Btk interacts specifically with the goblet cell apical membrane, which bounds the goblet cavity and contains the K+ pump responsible for the s.c.c . and high transepithelial potential difference (p.d.) . Such interaction results in a rapid disruption of K+ transport across the goblet cell apical membrane, leading to dissipation of the K+ gradient and loss of p.d . The histopathological changes previously reported by other workers would then be a consequence of K+ pump inhibition causing changes in the intracellular pH, Ca2+ etc . Some possible molecular bases for these specific interactions between Btk and cell membrane are discussed.

J Gen Microbiol, 1985 Mar, 131 ( Pt 3), 437 - 49
Effect of gramicidin S on the transcription system of the producer Bacillus brevis Nagano; Frangou-Lazaridis M et al.; The effect of the peptide antibiotic gramicidin S, produced by Bacillus brevis Nagano, was tested on the transcription system of the producer by using in vivo, semi in vitro and in vitro systems for studies of RNA synthesis . The effects of other peptide antibiotics (linear gramicidin, tyrocidine and tyrothricin) were also tested for comparison . It was found that (a) RNA polymerase isolated from either gramicidin S-producing or non-producing strains had a similar structure and requirements and that (b) the presence of gramicidin S caused a very strong inhibition of the in vitro transcription system . We present evidence that this inhibition is most probably through formation of a complex between the antibiotic and the DNA . In vivo studies indicate that transcription during growth and sporulation is not affected by gramicidin S and the implication is made that gramicidin S inhibits transcription during germination and outgrowth.

Mikrobiologiia, 1985 Mar-Apr, 54(2), 245 - 51
{Growth limitations and biosynthesis of gramicidin in Bacillus brevis var . G.-B}; Lykov VP et al.; Gramicidin S biosynthesis was studied in Bacillus brevis var . G.-B . during its batch and continuous cultivation when the culture growth was limited with nutrient sources (glycerol, ammonium nitrogen, phosphate), oxygen deficiency and the action of a physical factor (a low temperature) . The antibiotic biosynthesis was shown to be induced by a change in the growth rate caused by the action of any factor decelerating the growth . The authors propose a mathematical model for the antibiotic synthesis, biomass accumulation and the utilization of a substrate limiting the growth . The model is based on the age separation of cells . The model is analyzed in terms of optimizing the one-stage continuous cultivation process . The model allows one to calculate optimal conditions of the antibiotic synthesis in the process of one-stage continuous cultivation.

Biochemistry, 1985 Feb 26, 24(5), 1073 - 9
Purification and characterization of a membrane-associated phosphatidylserine synthase from Bacillus licheniformis; Dutt A et al.; A CDP-diacylglycerol-dependent phosphatidylserine synthase was solubilized from Bacillus licheniformis membranes and purified to near homogeneity . The purification procedure consisted of CDP-diacylglycerol-Sepharose affinity chromatography followed by substrate elution from blue dextran-Sepharose . The purified preparation showed a single band with an apparent relative molecular mass of 53 000 daltons when subjected to sodium dodecyl sulfate--polyacrylamide gel electrophoresis . Proteolytic digestion of the enzyme yielded a smaller (41 000 daltons) active form . The preparation was free of any phosphatidylglycerophosphate synthase, phosphatidylserine decarboxylase, CDP-diacylglycerol hydrolase, and phosphatidylserine hydrolase activities . The utilization of substrates and the formation of products occurred with the expected stoichiometry . Radioisotopic exchange patterns between related substrate and product pairs suggest a sequential Bi-Bi reaction as opposed to the ping-pong mechanism exhibited by the well-studied phosphatidylserine synthase of Escherichia coli {Larson, T . J., & Dowhan, W . (1976) Biochemistry 15, 5212-5218} . The B . licheniformis enzyme was also found to be markedly dissimilar to the E . coli enzyme with regard to association with detergent micelles, affinity for ribosomes, and antigenicity.

Nucleic Acids Res, 1985 Feb 25, 13(4), 1389 - 97
Does the 'non-coding' strand code?
Sharp PM.
The hypothesis that DNA strands complementary to the coding strand contain in phase coding sequences has been investigated . Statistical analysis of the 50 genes of bacteriophage T7 shows no significant correlation between patterns of codon usage on the coding and non-coding strands . In Bacillus and yeast genes the correlation observed is not different from that expected with random synonymous codon usage, while a high correlation seen in 52 E . coli genes can be explained in terms of an excess of RNY codons . A deficiency of UUA, CUA and UCA codons (complementary to termination) seems to be restricted to the E . coli genes, and may be due to low abundance of the relevant cognate tRNA species . Thus the analysis shows that the non-coding strand has the properties expected of a sequence complementary to a coding strand, with no indications that it encodes, or may have encoded, proteins.

FEBS Lett, 1985 Feb 25, 181(2), 313 - 7
Loss of liposome binding of NADH dehydrogenase from alkalophilic Bacillus on subtilisin digestion; Xuemin X et al.; Alkalophile NADH dehydrogenase consisting of two 65-kDa subunits was changed by subtilisin into an enzyme species consisting of two 38-kDa subunits . The amino acid composition and enzyme activity per molecule of the subtilisin-treated enzyme were almost the same as those of the native enzyme, respectively . On mixing with phospholipid liposome, the conformation of the native enzyme was changed, as suggested by the changes in the type of Arrhenius plot and of CD spectrum and enzyme activity . These conformational properties of the subtilisin-treated enzyme, on the other hand, were not affected by liposome . Gel filtration of the subtilisin-treated enzyme mixed with the liposome showed no binding of the protein to liposome.

Eur J Biochem, 1985 Feb 15, 147(1), 41 - 6
Structural studies on the linkage unit between poly(galactosylglycerol phosphate) and peptidoglycan in cell walls of Bacillus coagulans; Kaya S et al.; Structural studies were carried out on the linkage units in the teichoic-acid--glycopeptide complexes isolated from lysozyme digests of the cell walls of Bacillus coagulans AHU 1366 . On treatment with 47% hydrogen fluoride, the complexes gave a disaccharide characterized as glucosyl(beta 1----4)N-acetylglucosamine together with major fragments, galactosyl(alpha 1----2)glycerol . By means of Smith degradation and partial acid hydrolysis, the teichoic acid chain was shown to be composed of the repeating units, galactosyl(alpha 1----2)glycerol-3(1)-phosphate, which were joined by phosphodiester bonds at C-6 of the galactose residues . The mild alkaline hydrolysis of teichoic-acid-linked glycan fragments yielded teichoic acid chains and disaccharide-linked glycan fragments, from which the disaccharide, glucosyl(beta 1----4)N-acetylglucosamine, was liberated by mild acid hydrolysis, whereas the same disaccharide linked to the teichoic acid chain was obtained by direct heating of the cell walls at pH 2.5 . In addition, the presence of specialized glycerol phosphate units in the linkage region was shown by the isolation of tris(glycerol phosphate)3-glucosyl(beta 1----4)N-acetylglucosamine from the products of the Smith degradation of the teichoic-acid--glycopeptide complexes . Thus, it is concluded that the poly(galactosylglycerol phosphate) chain in the cell walls of B . coagulans AHU 1366 is linked to peptidoglycan through a novel linkage unit, bis(glycerol phosphate)-3-glucosyl(beta 1----4)N-acetylglucosamine.

Cancer, 1985 Feb 15, 55(4), 707 - 12
A randomized trial of adjuvant chemotherapy and immunotherapy in Stage I and Stage II cutaneous melanoma . An interim report; Sterchi JM et al.; Seventy patients with Stage I and II malignant melanoma were randomized to treatment with either intravenous dacarbazine alone or intravenous dacarbazine plus intradermal injection of the methanol-extracted residue of bacillus Calmette-Guerin (BCG) . Analysis of treatment failed to reveal a statistically significant difference between these two forms of treatment in Stage I and II patients . It is possible that chemoimmunotherapy may increase survival in Stage II patients, but this possibility should be interpreted with caution.

Nature, 1985 Feb 28-Mar 6, 313(6005), 803 - 6
Cloning and expression in Escherichia coli of the gene for human tumour necrosis factor; Shirai T et al.; Tumour necrosis factor (TNF) was found originally in mouse serum after intravenous injection of bacterial endotoxin into mice primed with viable Mycobacterium bovis, strain Bacillus Calmette-Guerin (BCG) . TNF-containing serum from mice is cytotoxic or cytostatic to a number of mouse and human transformed cell lines, but less or not toxic to normal cells in vitro . It causes necrosis of transplantable tumours in mice . TNF also occurs in serum of rat, rabbit and guinea pig . Rabbit TNF has been purified recently to give a single band on SDS-polyacrylamide gel electrophoresis (PAGE) . The purified TNF had a relative molecular mass (Mr) 40,000 +/- 5,000 measured by gel filtration, and 17,000 by SDS-PAGE . Its isoelectric point is 5.0 +/- 0.3 . The necrotic activity in vivo and the cytotoxicity in vitro are produced by the same substance . The gene encoding TNF has been identified in a human genomic DNA library using as a probe a cloned cDNA encoding a portion of rabbit TNF . The regions of this gene encoding an amino-acid sequence corresponding to mature TNF have been expressed in Escherichia coli and the product of this expression isolated in pure form and shown to produce necrosis of murine tumours in vivo.

Vet Immunol Immunopathol, 1985 Feb, 8(3), 225 - 44
Immune responsiveness and lymphoreticular morphology in cattle fed hypo- and hyperalimentative diets; Fiske RA et al.; Immune responses and lymphoreticular morphology were studied in 3 groups of yearling Hereford steers fed hypoalimentative, maintenance and hyperalimentative diets for 142 days . Significant decreases in plasma protein levels and circulating lymphocyte levels occurred in low energy intake steers . Percent circulating lymphocytes bearing surface immunoglobulins and serum levels of IgG and IgM did not vary significantly within or between groups . Antibody responses to Brucella abortus bacterin inoculated on day 63 were similar in all 3 groups . Antibody responses to chicken erythrocytes inoculated on day 88 were significantly lower in hypoalimentated steers . Differences between groups in lymphocyte blastogenic responses to phytohemagglutinin (PHA), concanavalin A (Con A) and pokeweed mitogen (PWM) were not significant . Hyperalimentated steers had significantly depressed PWM responses compared to a baseline value established for that group . In addition, hypoalimentated steers tended to have elevated responses to PHA, although differences were not significant . There were no significant differences between groups in dermal hypersensitivity responses to tuberculin following sensitization with viable Bacillus Calmette-Guerin (BCG) . The thymuses of hypoalimentated steers were markedly atrophied but lymph nodes and splenic white pulp were normal . Thymus, lymph nodes and spleen were normal in hyperalimentated steers.

Parasitology, 1985 Feb, 90 ( Pt 1), 101 - 10
Attempts to induce resistance to Schistosoma mansoni and S . haematobium in Kenyan baboons (Papio anubis) using non-specific immunostimulants; Sturrock RF et al.; Non-specific immunostimulants were used in an attempt to protect baboons from infection by schistosomes . Subcutaneous vaccination with cord factor (4.50 mg) and muramyl dipeptide (4.56 mg) 6 days before percutaneous exposure to 3000 Schistosoma haematobium cercariae/baboon (c.p.b.) failed to protect naive baboons: baboons with a 7-month-old, 5000 c.p.b . S . haematobium primary infection had developed too strong a natural immunity to detect any protection attributable to vaccination . Subcutaneous vaccination with 0.4 ml of Bacillus Calmette-Guerin (BCG, 1-8 X 10(8) colony forming units/ml) 4 days before exposure to 1000 c.p.b . S . mansoni gave a significant (38%) reduction in worm load compared with controls . However, vaccination with 0.8 (intramuscular) and 0.2 (intradermal) ml of BCG 11 days before exposure to S . mansoni 800 c.p.b . did not protect naive baboons, nor did it significantly reduce challenge worm recovery from baboons with a 13-week-old, 500 c.p.b . S . mansoni primary infection . Obvious pathology was seen at the site of vaccination in the first but not the second BCG experiment . These results partly support the findings in mice that non-specific macrophage and monocyte activators give partial protection against schistosome infections but they also illustrate that rodents and primates do not necessarily react identically . Hence, findings from rodent models should be extrapolated to man with some caution.

J Appl Bacteriol, 1985 Feb, 58(2), 207 - 14
Biological indicators for low temperature steam formaldehyde sterilization: effect of defined media on sporulation, germination index and moist heat resistance at 110 degrees C of Bacillus strains; Hoxey EV et al.; Choice of a biological indicator depends upon selecting a strain with the optimum balance of desirable properties . Screening 20 strains of Bacillus spp . for sporulation on three defined media has shown the wide variation that occurs in requirements for sporulation and properties of the resultant spores . Comparison of germination index and moist heat resistance of resultant spores suggest that a combination of high germination index, high heat resistance and linear inactivation may not be possible.

J Bacteriol, 1985 Feb, 161(2), 789 - 91
Mineralization and heat resistance of bacterial spores; Marquis RE et al.; The heat resistances of the fully demineralized H-form spores of Bacillus megaterium ATCC 19213, B . subtilis var . niger, and B . stearothermophilus ATCC 7953 were compared with those of vegetative cells and native spores to assess the components of resistance due to the mineral-free spore state, presumably mainly from dehydration of the spore core, and to mineralization . Mineralization greatly increased heat resistance at lower killing temperatures but appeared to have much less effect at higher ones.

Vet Immunol Immunopathol, 1985 Feb, 8(3), 273 - 87
Response of the lymph node to tumour antigen and Bacillus Calmette-Guerin in cattle bearing tumours of the ethmoturbinate region; Vikram Reddi M et al.; The immunological response of the Ln subiliacus (externus) in cattle bearing ethmoid carcinoma was studied . The lymph node was stimulated in vivo by I/D administration of tumour antigen and Bacillus Calmette-Guerin and the changes in the lymph node were studied at 3-day intervals up to 15 days . The histological changes in the lymph node were assessed and the functional activity of the activated macrophages in the lymph node impression smears was evaluated using Nitroblue tetrazolium salt reduction test . Similar studies were also undertaken in age-matched healthy nontumour bearing cattle . On stimulation with BCG and tumour antigen there was stimulation of the T cell dependent area . However, the response to the tumour antigen was quicker but shorter in duration compared with that to BCG . The response in healthy animals was proportionately much less . The studies indicated that tumour-bearing animals which were mostly in stage II were immunocompetent.

Urology, 1985 Feb, 25(2), 119 - 23
Experience with intravesical bacillus Calmette-Guèrin therapy of superficial bladder tumors; Herr HW et al.; Between March, 1978, and July, 1981, 86 patients with polychronotopic superficial papillary bladder tumors and concurrent carcinoma in situ were randomized to receive either transurethral resection alone (43) or TUR plus BCG (43) . The results indicate that BCG is not only active in preventing recurrences of new tumors but also effective for the diffuse, flat carcinoma in situ.

J Bacteriol, 1985 Feb, 161(2), 784 - 5
Molecular cloning and expression of the xylanase gene of alkalophilic Bacillus sp . strain C-125 in Escherichia coli; Honda H et al.; The gene for xylanase A of alkalophilic Bacillus sp . strain C-125 was cloned in Escherichia coli with pBR322 . The plasmid pCX311 contained 2.6- and 2.0-kilobase-pair HindIII fragments . The characteristics of the purified pCX311-encoded xylanase were the same as those of purified xylanase A from alkalophilic Bacillus sp . strain C-125.

Zhonghua Min Guo Wei Sheng Wu Ji Mian Yi Xue Za Zhi, 1985 Feb, 18(1), 44 - 53
{Extracellular thermostable alpha-amylase from Bacillus licheniformis B7}; Tan JA et al.; A thermostable alpha-amylase producing bacterium has been isolated from soil and identified as Bacillus licheniformis B7 . Cultural conditions of this strain for alpha-amylase production were studied . The optimum temperature and incubation period for enzyme production were 50 degrees C and 7 days respectively . Starch with a concentration of 2.5% was the best carbon source for enzyme production . Besides the above conditions, the addition of 0.05% FeCl3 X 6H2O, 0.6% yeast extract, 0.35% peptone, 0.2% Na-citrate and 0.008% CaCl2 X 2H2O to the growth medium produced maximum enzyme activity . The alpha-amylase produced by this strain showed maximal activity at 90 degrees C and pH 9.0.

Mol Cell Biochem, 1985 Feb, 66(1), 13 - 20
Active site studies on Bacillus amyloliquefaciens alpha-amylase (I); Dua RD et al.; Modification of liquefying alpha-amylase by diethylpyrocarbonate or its photo-oxidation in the presence of rose bengal caused rapid loss of enzyme activity . The photo-oxidation followed pseudo-first-order kinetics giving maximal value at pH 8.0 . The photo-oxidized enzyme showed a characteristic increase in absorbance at 250 nm which was directly proportional to the extent of inactivation . Diethylpyrocarbonate at low concentration at pH 6.0 and 30 degrees C completely inactivated alpha-amylase . Inactivation followed pseudo-first-order kinetics . The reaction order with respect to inactivation by diethylpyrocarbonate-modified enzyme showed increased absorbance at 240 nm which was reversed completely upon treatment with NH2OH at 30 degrees C for 16 hr . Calculating the histidine residues being modified from the increase in absorbance at 240 nm showed that three residues were ethoxyformylated on treatment with diethylpyrocarbonate, of which only one was found at the active site . Substrate and competitive inhibitor protects the enzyme against both, photo-oxidation, and modification by diethylpyrocarbonate, confirming that histidine plays an essential role at the alpha-amylase active site.

Proc Natl Acad Sci U S A, 1985 Feb, 82(4), 1035 - 9
Two alkaline phosphatase genes positioned in tandem in Bacillus licheniformis MC14 require different RNA polymerase holoenzymes for transcription; Hulett FM et al.; Southern transfer analysis of Bacillus licheniformis MC14 DNA, using as probe a DNA fragment from within the coding region of a previously cloned alkaline phosphatase (APase) gene, revealed a second area of hybridization adjacent to the cloned APase gene . A second APase gene (APase II) was subcloned from the same plasmid clone, pMH8, from which the first APase gene (APase I) had been subcloned . The two genes are arranged in tandem with several hundred base pairs separating them . Immunoblot analysis showed that both code for Mr 60,000 proteins that crossreact with anti-APase . Both proteins enzymatically cleave 5-bromo-4-chloro-3-indolyl phosphate . In vitro transcription showed that APase I and APase II are transcribed in the same direction but that the two genes require different forms of Bacillus RNA polymerase: sigma 55- and sigma 37-containing RNA polymerase holoenzymes, respectively.

Biochem Biophys Res Commun, 1985 Jan 31, 126(2), 966 - 73
An active center tryptophan residue in liquefying alpha-amylase from Bacillus amyloliquefaciens; Kochhar S et al.; Liquefying alpha-amylase from Bacillus amyloliquefaciens was inactivated on treatment with N-bromosuccinamide . Preincubation of the enzyme with either of the substrate, or competitive inhibitor provided significant protection against inactivation . The relationship between activity loss and the number of tryptophan residues modified, as well as presence of substrate/inhibitor in the reaction mixture, demonstrated that only one of three modifiable tryptophan residues is at or near the active center . The apparent Km of the modified enzyme for soluble starch increased manifold, thus implicating the sensitive tryptophan residue in the substrate binding region of the enzyme.

Biochem Biophys Res Commun, 1985 Jan 31, 126(2), 961 - 5
Separation of the cytolytic and mosquitocidal proteins of Bacillus thuringiensis subsp . israelensis; Hurley JM et al.; The cytolytic and mosquitocidal proteins of Bacillus thuringiensis subsp . israelensis were isolated from parasporal crystals and subsequently separated from each other . The proteins were separated by gel filtration chromatography and their molecular weights were estimated by both gel filtration chromatography and SDS-polyacrylamide gel electrophoresis . The apparent molecular weights of the mosquitocidal protein and the cytolytic protein were estimated to be 65,000 daltons and 28,000 daltons, respectively.

Biochem Biophys Res Commun, 1985 Jan 31, 126(2), 953 - 60
Diversity of protein inclusion bodies and identification of mosquitocidal protein in Bacillus thuringiensis subsp . israelensis; Lee SG et al.; Bacillus thuringiensis subsp . israelensis produces, during sporulation, protein inclusion bodies of wide ranging sizes, all of which are toxic to mosquitoes . Two proteins are present in the smallest protein bodies (less than 0.2 micron dia.), but the number of proteins increases with increasing size of protein bodies . The largest bodies (greater than 1.5 micron dia.) contain seven proteins . All of the proteins are synthesized at different times during sporulation and are added to developing protein bodies in a stepwise manner . The protein component responsible for mosquitocidal activity is a 65,000-dalton protein, that is present in all of the protein bodies.

Schweiz Med Wochenschr, 1985 Jan 19, 115(3), 93 - 6
{Aminoglycoside levels in bronchial secretions}; Mombelli G; Aminoglycosides are of limited clinical efficacy in gram-negative bacillary pneumonia, although they are commonly employed to treat this infection . This poor efficacy has been related in part to host factors (abnormalities in the lung, immunocompromise) and in part to pharmacologic factors . In particular, aminoglycoside levels in bronchial secretions are often borderline or inadequate in relation to the minimal inhibitory concentrations for most gram-negative strains . The low aminoglycoside concentrations result from poor penetrance into the respiratory tract or from local inactivation of these drugs, but basically reflect their low therapeutic-to-toxic ratio . The endotracheal injection of aminoglycosides resulted in high bactericidal activity within the bronchial lumen and in increased clinical efficacy, without increasing systemic toxicity . In view of the potential dangers of topical antibiotics, however, endotracheal treatment should be confined to selected patients.

Can Med Assoc J, 1985 Jan 15, 132(2), 137 - 40
Tuberculosis in female nurses in British Columbia: implications for control programs; Burrill D et al.; All 57 cases of active tuberculosis in women in nursing and related assisting occupations (henceforth called nurses) notified in British Columbia between 1969 and 1979 were reviewed . This represented a mean annual incidence of active tuberculosis of 2.6/10 000, similar to that in other women, adjusted for age and birthplace . The rate varied according to birthplace: among nurses born in Canada the rate was 2.0, almost twice that of other women born in Canada, and among those born in Asia it was 24.8, less than half that of other women born in Asia . The nurses born in Canada who had received BCG (bacille Calmette-Guerin) during their training were least likely to contract tuberculosis, the incidence rate being comparable to that among other women . Those whose results of tuberculin testing were negative but who were not vaccinated were twice as likely to contract tuberculosis, whereas those whose results were positive at the start of training were four times as likely to contract tuberculosis . The feasibility and implications of a tuberculosis screening and surveillance program are discussed.

J Biol Chem, 1985 Jan 10, 260(1), 616 - 23
Fatty acid biosynthesis in Mycobacterium tuberculosis var . bovis Bacillus Calmette-Guérin . Purification and characterization of a novel fatty acid synthase, mycocerosic acid synthase, which elongates n-fatty acyl-CoA with methylmalonyl-CoA; Rainwater DL et al.; A crude extract from Mycobacterium tuberculosis var . bovis Bacillus Calmette-Guerin was previously shown to incorporate methylmalonyl-CoA into mycocerosic acids, exemplified by 2,4,6,8-tetramethyloctacosanoic acid, and malonyl-CoA into n-fatty acids (Rainwater D . L., and Kolattukudy, P . E . (1983) J . Biol . Chem . 258, 2979-2985) . The presence of several fatty acid synthases with differences in substrate preference and product chain length was detected in the crude extract of M . tuberculosis var . bovis . Among them was a mycocerosic acid synthase which was purified to homogeneity using anion-exchange chromatography, gel filtration, affinity chromatography, and hydroxylapatite chromatography . This fatty acid synthase elongated long-chain fatty acyl-CoA primers using methylmalonyl-CoA and NADPH to produce multimethyl-branched mycocerosic acids . The enzyme was specific for methylmalonyl-CoA and would not incorporate malonyl-CoA into fatty acids . It elongated n-C6 to n-C20 CoA esters to generate primarily the corresponding tetramethyl-branched mycocerosic acids . Exogenous {1-14C}acyl-CoA and trideuteromethylmalonyl-CoA were incorporated into the multimethyl-branched fatty acids . Dodecyl sulfate electrophoresis showed that the enzyme had a molecular weight of 238,000, whereas gel filtration showed a native molecular weight of 490,000, indicating that the enzyme is composed of two monomers of identical molecular weight . The enzyme contained an acyl carrier protein-like segment as indicated by incorporation of {1-14C} pantothenate into the 238-kDa protein and production of 1 mol of taurine/mol of the monomer upon hydrolysis of performic acid-oxidized enzyme . It is concluded that the mycocerosic acid synthase is a multifunctional enzyme similar to the well-characterized multifunctional fatty acid synthases except for the substrate specificity.

J Mol Biol, 1985 Jan 5, 181(1), 145 - 6
Crystallization and preliminary X-ray data for the exocellular beta-lactamase of Bacillus licheniformis 749/C; Dideberg O et al.; The exocellular beta-lactamase from Bacillus licheniformis 749/C has been crystallized from polyethylene glycol solution at pH 5.5 . An X-ray examination of the monoclinic crystals shows the space group is P21, with unit cell dimensions a = 66.77 A, b = 93.77 A, c = 43.57 A and beta = 104.5 degrees . The asymmetric unit consists of two molecules of 28,500 Mr each . The crystals are suitable for structure analysis to at least 2 A resolution.

IARC Sci Publ, 1985, (60), 375 - 82
Treatment of Burkitt's lymphoma: the African experience; Olweny CL et al.; Although Burkitt's lymphoma (BL) can be treated by surgery, radiotherapy and immunotherapy, chemotherapy is the mainstay of treatment . This paper summarizes the various clinical trials undertaken in Africa over the past decade . The single most effective drug for BL is cyclophosphamide (CPM) . Given alone for remission induction, CPM is as effective as combinations consisting of either CPM, vincristine (VCR) and methotrexate (MTX) or CPM, VCR and cytosine arabinoside (Ara-C) . Survival data indicate that single-dose CPM is comparable to multiple doses . Thus, maintenance therapy may not be necessary, and may in fact worsen the final outcome . Intrathecal (IT) MTX given together with systemic therapy significantly delays central nervous system (CNS) relapse, which is not prevented by cerebrospinal irradiation . For established CNS disease, IT-Ara-C for three days followed by MTX on the fourth day is effective . Bacillus Calmette-Guerin scarification, while provoking measurable responses in vivo and in vitro, had no measurable, specific anti-tumour reaction, since no effect was observed on relapse rate, duration of remission or survival . High-dose CPM produces objective responses in patients previously resistant to conventional doses . Teniposide (VM 26) is currently undergoing phase 2 trial, and definite short-lived responses have been recorded.

Geogr Med, 1985, 15, 1 - 16
Tuberculosis epidemiological situation in the world and in Hungary (1882-1982); Nemeth T et al.; The centenary of the discovery of the tuberc bacillus (Koch-centenary) threw light on the unsolved problems of the epidemiology of tuberculosis in the world . Despite of the definitively significant progress made since the turn of the century the world-wide elimination and eradication of tuberculosis is not yet accomplished . Deficiencies in the fight against tuberculosis in the developing countries which represent two-third of the world play firstly a role in it . The number of new tuberculosis cases keeps being about 5.000 yearly in Hungary thus the present programme requires modification . An increased fight against tuberculosis on a global scale may be successful only with international collaboration and with higher rate of financial support provided by the highly developed industrial countries.

Cancer Drug Deliv, 1985 Spring, 2(2), 127 - 32
Enhanced antitumor effects with intralymphatic delivery using bacillus Calmette-Guerin in animal models; Jeglum KA et al.; This study compares the effectiveness of various routes of administration of Bacillus Calmette-Guerin (BCG) utilizing the New Zealand white (NZW) rabbit V2 carcinoma model . The routes compared were intratumor, intravenous, scarification, subcutaneous, and intralymphatic . Primary tumor regression, disease-free interval, and survival were measured . The disease-free interval and survival for the intralymphatic group were significantly longer (p less than 0.05) than all groups except the scarification group . That group had a prolonged survival as compared with all groups except the intralymphatic group . The intralymphatic route of administration was the most effective method in causing local tumor regression and curtailing metastasis . This study clearly demonstrates that the intralymphatic route requires further mechanistic studies and clinical investigation as a means for delivering biological response modifiers.

Bull Pan Am Health Organ, 1985, 19(2), 206 - 8
The epidemiology of tuberculosis in Chile; Heliobacterium and the origin of chrysoplasts; Chrysoplasts, golden-yellow and brown photosynthetic membrane-bounded plastids, photosynthetic organelles of algae such as phaeophytes (brown seaweeds), bacillariophytes (diatoms) and chrysophytes (golden-yellow algae including silicoflagellates), are hypothesized to have originated from brownish photoheterotrophic bacteria such as the newly discovered anaerobic nitrogen-fixing Heliobacterium . The consequences of this hypothesis as well as the data required to verify or disprove it are presented.

Arkh Patol, 1985, 47(7), 34 - 41
{Myocardial morphology in endotoxic shock}; Iakovlev MIu; The myocardium of rabbits with endotoxin shock produced by two intravenous injections (0.1 and 0.3 mg/kg) of the typhoid fever bacillus antigen was studied histologically, histochemically, histoenzymatically and by polarization microscopy . Animals were killed by decapitation 0,5-1,5, 5-10, 24-30, 48-80, 120-194 hours after the 2nd injection; animals dying in the course of experiment were also examined . Morphological substrate of heart insufficiency, morphofunctional heterogeneity of cardiomyocytes of both inner and external myocardial layers were revealed . Hypoxia of a complex origin is the main cause of the above alterations.

Intervirology, 1985, 24(1), 33 - 41
Assembly and accumulation sites of maize mosaic virus in its planthopper vector; Ammar ED et al.; The morphology, assembly, and accumulation sites of rhabdovirus particles in Peregrinus maidis planthoppers infected with a Hawaiian isolate of maize mosaic virus (MMV) were studied . These particles were usually bullet-shaped, but were sometimes bacilliform, and averaged 234 and 247 nm, respectively, in length and 60 nm in width . They were found in most acini of the principal and accessory salivary glands and in brain, nerve ganglia, leg muscle, foregut, midgut, trachea, epidermis, and fat and connective tissues . In most tissues MMV particles accumulated mainly within intracytoplasmic, dilated cisternae that were connected to the perinuclear space . However, in the salivary glands virus particles accumulated mainly in intercellular and extracellular spaces and were found in secretion vesicles . MMV particles appeared to bud from three types of membranes: (i) inner, and rarely outer, nuclear membranes of cells in most tissues examined; (ii) intracytoplasmic membranes, e.g., endoplasmic reticulum in salivary glands; and (iii) plasma membranes of salivary gland cells and nerve axons . The plasma membrane has not been reported previously as a budding site for plant rhabdoviruses, although it is known as a major assembly site for animal rhabdoviruses.

Folia Parasitol (Praha), 1985, 32(3), 271 - 7
The efficacy of Bacillus thuringiensis var . israelensis against larvae of the blackfly Odagmia ornata (Meig.) (Simuliidae) at low temperatures; Olejnicek J et al.; The effect of the suspension of Bacillus thuringiensis var . israelensis spores on larvae of the blackfly Odagmia ornata was studied in the laboratory and under field conditions of a natural biotope in southern Bohemia . The preparation Moskitur was used and its effect was tested in laboratory at temperatures 0.1-2.9 degrees C and 17-19 degrees C . Although O . ornata larvae were able to filter feed on a lethal dose of the preparation even at a lower temperature than 2.9 degrees C, no marked manifestation of mortality was observed at low temperatures in comparison with a control sample.

Acta Microbiol Hung, 1985, 32(1), 65 - 73
Partial purification and some properties of tryptophan decarboxylase from a Bacillus strain; Buki KG et al.; Bacteria of different origin were screened for tryptophan decarboxylase activity . The best producer belonged to an unidentified taxonomic entity of the genus Bacillus . In complete medium it produced tryptamine from tryptophan . The decarboxylase could partially be purified from the cells by sonication and DEAE-cellulose chromatography . The enzyme had an Mr of 150 000 and a pH optimum of about 7, was stable up to 37 degrees C, and its Km was about 0.3 mM for tryptophan . The enzyme needed pyridoxal phosphate for maximum activity.

Folia Microbiol (Praha), 1985, 30(4), 342 - 8
Enhancement of bacitracin biosynthesis by branched-chain amino acids in a regulatory mutant of Bacillus licheniformis; Supek V et al.; DL-4-Azaleucine-resistant mutant of Bacillus licheniformis azlr-1 isolated after N-methyl-N'-nitro-N-nitrosoguanidine mutagenesis, was a better bacitracin producer than the parent strain . In the minimal medium, the antibiotic biosynthesis was 4 times higher in the mutant than in the parent strain and less dependent on L-leucine addition . In the complex fermentation medium, the yield was 18-20% higher in the mutant strain . Transaminase B activity measured in the crude extract revealed that the branched-chain amino acid biosynthetic enzymes were 5-10 times derepressed supplying bacitracin synthetase with enhanced quantity of isoleucine and leucine, the building units of bacitracin molecule.

Drugs, 1985, 29 Suppl 5, 162 - 6
A study of the penetration of temocillin in the cerebrospinal fluid; Bruckner O et al.; Temocillin concentrations in serum and cerebrospinal fluid (CSF) samples from 8 patients were assayed by high pressure liquid chromatography . It was possible to determine sequential series of ventricular (CSF) and serum concentrations in 4 adult neurosurgical patients with slight to moderate impairment of blood-CSF barriers, because serial CSF samples were obtained from external ventricular drains . In 4 other patients with meningitis, temocillin was given in addition to the regular treatment schedule and 6 'spot' concentrations of temocillin in lumbar CSF and serum samples were determined . Temocillin CSF concentrations in these subjects seemed to be inadequate for the treatment of Gram-positive bacterial meningitis and only partially valuable for the treatment of Gram-negative bacillary meningitis.

Nauchnye Doki Vyss Shkoly Biol Nauki, 1985, (6), 89 - 91
{Effect of bacitracin on the sporulation of Bacillus licheniformis 28 KA}; Egorov NS et al.; The bacitracin effect on spore germination in Bacillus licheniformis 28 KA--the producer of this antibiotic on media of various composition has been studied . Different intensity of antibiotic synthesis has been revealed on various media . A high content of presynthesized bacitracin has an inhibitory effect on spore germination in B . licheniformis.

Tissue Cell, 1985, 17(3), 379 - 94
Effect of sublethal Bacillus thuringiensis crystal endotoxin treatment on the larval midgut of a moth, Manduca: SEM study; Spies AG et al.; The effect of a single, sublethal dose of B . thuringiensis crystal endotoxin on the midgut of the moth Manduca sexta larvae was monitored during acute and recovery stages . Initially both goblet and columnar cells swelled . Many columnar cells produced membrane extrusions . In some cases the affected cells ruptured, extruding cellular debris into the midgut lumen . Following the acute stage, the midgut tissue recovered, the damaged cells being extruded into the midgut lumen apparently as newly regenerated cells rose to take their place . The insects appeared to recover completely and continue normal development.

Respiration, 1985, 47(4), 285 - 92
Paradoxical functions of alveolar macrophages from Calmette-Guérin bacillus-immunized rats; Shiratori Y et al.; The effect of autologous alveolar macrophages on the lymphoproliferative response to mitogens was investigated using Calmette-Guerin bacillus (BCG)-immunized rats . The proliferation of lymphocytes obtained from normal rats was suppressed when the lymphocytes were cultured with alveolar macrophages at any proportion, but there was an enhancement in the lymphoproliferative response when the lymphocytes from BCG-immunized rats were incubated with alveolar macrophages at the ratio of 100:1 . The supernatant of alveolar macrophage cultures from both normal and BCG-immunized rats showed a suppressive effect on lymphocytes while the supernatant of lymphocyte-alveolar macrophage cultures from BCG-immunized rats enhanced the lymphoproliferation . The results indicate that there are soluble factors elaborated by alveolar macrophages which have opposite effects on lymphoproliferation, and that these factors may be involved in the development of the hyperimmune state of the lung in tuberculosis.

Oncology, 1985, 42(4), 259 - 64
Comparative studies between liposomes containing muramyl dipeptide and various immunomodulators on activation of mouse peritoneal macrophages and NK cells; Sakita M et al.; We compared the effects among muramyl dipeptide (MDP), liposome-encapsulated MDP (liposome MDP), bacillus Calmette-Guerin (BCG) and OK-432 on cytotoxic activity of mouse peritoneal macrophages (PM) and natural killer (NK) cells in vitro and in vivo, and their tumor-inhibitory effects against MH134 ascitic tumors in C3H/He mice . The cytotoxicity of PM induced by free MDP was lower than that induced by BCG, but a significantly higher cytotoxicity was induced by liposomes containing MDP and OK-432 . The peritoneal NK cells were not activated by MDP, liposome MDP or BCG, but OK-432 profoundly augmented peritoneal NK activity . Growth inhibition of ascitic tumor was not observed in free MDP and BCG intraperitoneally treated mice, but moderate growth inhibition was noted in liposome-MDP-treated mice; and in OK-432-treated mice, marked tumor growth inhibition and prolongation of survival time were observed . These results suggested that OK-432 is more advantageous in controlling malignant tumor growth in vivo than free MDP, liposome MDP or BCG because of its ability to activate both macrophages and NK cells.

Ann Inst Pasteur Microbiol, 1985 Jan-Feb, 136A(1), 91 - 8
How does a Bacillus split its septum right down the middle?
Koch AL, Kirchner G, Doyle RJ, Burdett ID.
This is a speculative paper which considers the possible ways that Gram-positive cells might employ to achieve an even thickness of the two daughter poles resulting from the fission of the cross-wall . The first is that the protonmotive force generated by the extrusion of protons at the cytoplasmic membrane acts to inhibit autolysins to a distance of about 25 nm . The second has to do with the stresses that develop as the poles form . On the tacit assumption that the autolysins will function faster when their substrates are under tension, it is shown how this, too, can lead to even bisection of the cross-wall . These possibilities are not alternative, both probably function.

Toxicon, 1985, 23(1), 145 - 55
Physiological actions of phosphatidylinositol-specific phospholipase C from Bacillus thuringiensis on KB III cells: alkaline phosphatase release and growth inhibition; Ikezawa H et al.; Phosphatidylinositol-specific phospholipase C of Bacillus thuringiensis caused the release of alkaline phosphatase from KB III cells and plasma membrane preparations prepared from the cells . Phosphatidylinositol-specific phospholipase C added to the culture of KB III cells inhibited cell growth by 30% . The release of alkaline phosphatase induced by phospholipase C was dependent on, or proportional to, the reaction time and the concentrations of the phospholipase C, KB III cells and plasma membrane preparation . The Arrhenius plot for phosphatase-release reaction showed a single break at 18.1 degrees C for KB III cells or at 27.3 degrees C for the plasma membrane preparation . The activation energies of the phosphatase-release reaction were 3.2 and 29.2 kcal/mol for KB III cells, and 4.3 and 26.5 kcal/mol for the plasma membrane preparation . The released alkaline phosphatase had a mol . wt of 105,000 and isoelectric points of 4.05 (major) and 4.4 (minor).

Pharmacology, 1985, 30(4), 181 - 7
Immunologic and anti-immunosuppressive effects of vitamin A; Nuwayri-Salti N et al.; The effects of vitamin A alcohol on cell-mediated immunity in vitro and its ability to prevent the immunosuppressive effects of prednisolone and cyclophosphamide in vivo were studied in mice . Lymphocytes from Calmette-Guerin bacillus (BCG) sensitized mice were stimulated specifically with purified protein derivative (PPD) and nonspecifically with phytohemagglutinin (PHA) . In the vitamin A injected animals there was significant enhancement of the spleen lymphocyte transformation not only in the PPD-sensitive cells but also in the T cells at large . In addition, vitamin A was able to restore to normal the cellular and humoral forms of immunity in prednisolone and cyclophosphamide-treated animals . It is suggested that vitamin A in nontoxic doses may have a role in enhancing the responses to weak immunogens and in reversing immunosuppression.

Jpn J Antibiot, 1985 Jan, 38(1), 69 - 73
{Microbial flora of the cervix from rupture of the membranes and the antibacterial effect of amniotic fluid}; Chimura T et al.; The endocervical flora was examined in 20 with rupture of membranes, 20 pregnant women served as control . The isolated organisms were aerobic and anaerobic in both groups, whereas P . cepacia was common in group of rupture of membranes, but another microorganisms were not statistically significant . Culture specimens taken from the vagina in nonpregnant women showed a higher presence of total bacteria than did those in pregnant women . The MICs of ampicillin (ABPC) were measured against isolated aerobic organisms from rupture of membranes . The distribution of sensitivity of Bacillus sp . was 0.20 microgram/ml, but another organisms were inhibited in concentration of higher than 3.13 micrograms/ml . The antibacterial effect of amniotic fluid during 24 weeks of gestation on S . aureus and S . pyogenes, but during 34 to 39 weeks of gestation the amniotic fluid has inhibitory effect only on the growth of K . pneumoniae . In contrast, the growth of E . coli was not inhibited by amniotic fluid during 24 to 39 weeks of gestation.

Bull Soc Pathol Exot Filiales, 1985, 78(1), 15 - 27
{Epidemiologic aspects of tuberculosis in Yaounde (Republic of Cameroon)}; Heyraud JD et al.; A retrospective investigation was carried out in March 1984 on files of 447 tubercular patients traced in 1982 in the Unit of Pneumophthisiology of the Jamot Hospital in Yaounde . It demonstrates that: 80% of the patients have a pulmonary localization, 95% of the pulmonary tuberculosis are diagnosed by bacilloscopy often after a long latency, 60% of two-sided forms and 64% of forms with excavation, by radiological examination, 65% of the recorded patients live outside Yaounde, a one year standard treatment follow-up leads to 50% cures, 50% of the patients do not come back before six months and 50% after six months . The epidemiological findings done from this investigation lead to suggest adaptations of the fight-programme relevant to the bacilloscopic tracing, the curative protocols and the BCG vaccination.

Arch Fr Pediatr, 1985 Jan, 42(1), 35 - 6
{Acute endocarditis caused by Kingella kingae in an infant}; La Selve H et al.; A case of bacterial endocarditis in a one year-old boy is reported . There was no underlying heart disease . The organism was a Kingella kingae, an aerobic Gram negative bacillus, a normal inhabitant of the upper respiratory tract . It has rarely been implicated as a pediatric pathogen . Occasionally it can cause bone and joint infections and exceptionally endocarditis.

J Med Chem, 1985 Jan, 28(1), 3 - 9
1H-2-Benzopyran-1-one derivatives, microbial products with pharmacological activity . Conversion into orally active derivatives with antiinflammatory and antiulcer activities; Shimojima Y et al.; A novel gastroprotective substance, 6-{{1(S)-{3(S),4-dihydro-8-hydroxy-1-oxo-1H-2-benzopyran-3-yl} -3-methylbutyl}amino}-4(S),5(S)-dihydroxy-6-oxo-3(S)-ammoniohexanoate (AI-77-B, 1), isolated from a culture broth of Bacillus pumilus AI-77, was chemically modified to prodrugs that are active by oral dosing . Compound 1 was lactonized and then monoalkylated at the primary amine position . Six N-alkylated gamma-lactone derivatives of 1 (with alkyl chains being methyl 5a, ethyl 5b, n-propyl 5c, n-butyl 5d, n-pentyl 5e, or n-hexyl 5f) were synthesized and eight compounds including 1 and gamma-lactone derivative 2 were compared for their gastroprotective activities and blood levels after oral administration in rats . Further, chloroform-water partition coefficients of 5a-f were also compared as a measure of lipid solubility . The protective effects of these compounds on stress ulcers were mutually related to blood levels of dealkylated compounds (1 and 2) . Parent compound 1 was detected in blood at 1 h after each of 5a-d was administered . When 5b or 5c was administered, high activity and high blood levels of 1 were observed in comparison with those levels obtained with 5a or 5d . Neither 5e nor 5f were detected in any amount in blood by oral administration without special formulation due to extremely low solubilities and agglutinative properties in intestinal fluid . Interestingly, 5b and 5c were found to have antiinflammatory activities in addition to potent antiulcerogenicity action.

Infect Immun, 1985 Jan, 47(1), 294 - 300
Enhancement of experimental bacteremia and endocarditis caused by dysgonic fermenter (DF-2) bacterium after treatment with methylprednisolone and after splenectomy; Butler T et al.; The dysgonic fermenter-2 bacterium is a newly recognized fastidious gram-negative bacillus that causes bacteremia and sometimes endocarditis in immunocompromised persons after they are bitten by dogs . To develop an experimental model of this infection, we placed polyethylene catheters across the aortic valves of New Zealand white rabbits, which were inoculated intravenously the next day with dysgonic fermenter-2 bacteria . After 1 week, the rabbits were killed and the endocardial vegetations were homogenized for quantitative culture . Large inocula (1.3 X 10(10) to 2.1 X 10(10) viable bacteria) were required to produce infected vegetations . All infected rabbits had negative blood cultures at the time of autopsy and most developed serum agglutinins against dysgonic fermenter-2 bacteria . Three daily injections of methylprednisolone (30 mg/kg), starting the day before inoculation, significantly increased the incidence of endocarditis and the number of bacteria per gram of infected vegetation (P less than 0.05) . Treatment with methylprednisolone prolonged the initial bacteremia and caused significant increases in the numbers of bacteria per gram of blood, spleen, and liver compared with those of untreated controls (P less than 0.05) . Rabbits that had previously undergone splenectomy showed prolongation of the initial bacteremia but no significant increase in the incidence of infected vegetations . These results showed that the dysgonic fermenter-2 bacterium is a pathogen that causes endocarditis in rabbits but that it requires a large inoculum and produces blood culture-negative infections . Treatment with methylprednisolone enhances infection by prolonging the initial bacteremia and probably by diminishing bactericidal activity in the vegetations.

Microbiol Immunol, 1985, 29(12), 1151 - 62
Synthesis and deposition of spore coat proteins during sporulation of Bacillus megaterium; Imagawa M et al.; Rabbit (anti-spore coat protein) IgG was prepared by immunization with coat proteins extracted with sodium dodecyl sulfate and dithiothreitol from isolated spore coats of Bacillus megaterium ATCC 12872 . Coat proteins were detected from 3 hr after the end of exponential growth (t3) in the mother cell cytoplasmic fraction by sandwich enzyme immunoassay using this antibody . The proteins in the forespore coat protein fraction increased from t3 and reached a plateau at t10 . Immunoblot analysis for the coat proteins in sporulating cells revealed the sequential synthesis of various proteins in the mother cell cytoplasmic fraction and simultaneous deposition of the same proteins as in the forespore coat fraction . These results suggest that turnover of precursor proteins of the spore coat is very rapid if precursor proteins are produced and they are proteolytically processed to produce mature proteins . Specific antibody to the 48,000-dalton protein, which is a major protein, did not cross-react with any other major (36,000, 22,000, 19,500, and 17,500-dalton) proteins . Specific antibody to the 22,000-dalton protein did not cross-react with the 48,000, 36,000, 19,500, 17,500, and 16,000-dalton proteins, but did cross-react with the 44,000, 25,000, and 12,000-dalton proteins.

Microbiol Immunol, 1985, 29(12), 1139 - 49
Germination of the decoated spores of Bacillus megaterium; Nakatani Y et al.; Decoated spores of Bacillus megaterium ATCC 12872 were prepared by extracting the inner coat components with an alkaline solution containing sodium dodecyl sulfate and dithiothreitol (SDS-DTT) from outer coat-deficient mutant spores, which were produced from one of the mutants isolated and named MAE-05 by us . The decoated mutant spores germinated as well as the intact spores of the mutant and the parent, indicating that the outer and inner spore cats cannot be essential structures for the initiation of germination . When the SDS-DTT-treated MAE-05 spores were converted to H-spores by incubation in citrate-phosphate buffer (pH 3.5) at 30 C for 3 hr, they lost their germinability by glucose and KNO3 . Ca-spores, prepared by treating H-spores with 10 mM calcium acetate at 37 C for 60 min, regained the germinability . Experiments on the interaction of 45Ca with the cortex and the inner membrane isolated from H-spores suggested that the calcium present in the inner membrane might be related to germinability.

Microbios, 1985, 44(177), 33 - 44
Mechanism of Ca2+ and dipicolinic acid requirement for L-alanine induced germination of Bacillus cereus BIS-59 spores; Kamat AS et al.; Spores prepared from different sporulating media containing varying amounts of Ca and dipicolinic acid (DPA), exhibited differential responses to germination in L-alanine (0.25 M) . Ca-spores with moderately high Ca and DPA contents could be triggered to germination by L-alanine, whereas P-spores with low contents of Ca and DPA could not be germinated by L-alanine unless Ca2+ or DPA was exogenously added . The initiation of L-alanine induced germination by P-spores in the presence of 45CaCl2 was associated with a marked uptake of 45Ca2+ . Experiments involving stepwise extraction of 45Ca from prelabelled spores indicated that a part of the spore calcium may be involved in L-alanine induced germination . Both Ca2+ and DPA seemed to have a stimulatory effect on the incorporation of 14C-L-alanine.

Mem Inst Oswaldo Cruz, 1985 Jan-Mar, 80(1), 1 - 9
{Evaluation of the incidence and toxicity of samples of Bacillus cereus in various classes of foods sold and consumed in the State of Rio de Janeiro}; Rabinovitch L et al.; One hundred and fourteen strains of Bacillus cereus were isolated during presumptive plate-counts from 18 groups of industrialized, non-industrialized, crude or cooked food, belonging to 10 separate classes . Specific presumptive counts ranged from 10(2) to 6 X 10(3)/g or ml . Among these isolates, 13 strains were derived from 3 outbreaks of food poisoning (involving a minimum of 57 people), as determined by the assayed bacteriological quality of the ingested foods . As an adopted procedure to correlate toxicity and ability to promote illness in man, culture fluids of all strains were assayed to determine their ability to increase vascular permeability (APC) to cause necrosis in rabbits skin and to kill albino mice . APC was positive in 86.85% of the 114 strains, death of albino mice occurred in 65.79% and a combination of APC and death was observed in 59.65% . APC plus necrosis, or only necrosis, occurred with 34.21% of the culture fluids . Death, APC and death with or without necrosis, were demonstrated in the strains implicated with illness . This confirmed the known individuality of action exhibited by certain B . cereus food-borne toxigenic factors . The low presumptive counts of this bacterium in the order of 10(2)-10(3)/g or ml found in food, implicated or not with illness, suggests that the recommended number of B . cereus per g or ml of food sample should be reevaluated in our country . Furthermore, a wider range of food should be brought under bacteriological sanitary control for this species.

Microbiol Immunol, 1985, 29(8), 689 - 99
Collapse of cortex expansion during germination of Bacillus megaterium spores; Nakatani Y et al.; When spores of Bacillus megaterium ATCC 12872 were incubated with CdCl2, they germinated without decomposition of the cortex . Moreover, the volume ratio of cortex to protoplast-plus-cortex, C/(P+C), of the CdCl2-germinated spores was reduced . Incubation of isolated cortex with the divalent compounds Cd2+, Ca2+, and Mg2+ reduced the gel volume to about 1/5 but incubation with a nonionic compounds, glucose, did not . The spores with reduced C/(P+C) were observed in the early period of glucose-induced germination . The time required for a 50% change in cortex morphology to occur was 2.5 min, which corresponds well with the time for 50% loss of heat resistance . This time was shorter than that necessary for release of peptidoglycan fragments and hydrolysis of cortex glycan chains . These data indicate that cortex hydrolysis is not related to the initiation of germination . 50% of the dipicolinic acid, calcium and magnesium were released at 3.4, 4.0, and 2.4 min, respectively . These results suggest that collapse of cortex expansion by the interaction of cortex with dipicolinic acid and cations released from the core, or exogenous ionic germinants is an important step in the initiation of germination.

Gene, 1985, 37(1-3), 125 - 30
Nucleotide sequence analysis of DNA replication origins of the small Bacillus bacteriophages: evolutionary relationships; Yoshikawa H et al.; The ends of the small Bacillus phage genomes serve as origins and termini of their DNA replication . We have determined nucleotide sequences at the termini of four different phage DNAs and compared them with those of phi 29 DNA which has been described previously . A high degree of homology was found at the extreme ends of DNAs from phi 29, phi 15 (group A), M2Y and Nf (group B) . 17 bp at the far left of the DNAs are identical . A highly conserved dodecanucleotide sequence, CCATTTCCCCAT, was also found in the righthand terminus of all these phage DNAs, at positions 27-38 from the end . Nucleotide sequences of phage GA-1 are not very similar to those of the other phages . Examination of the 5'-terminal and 3'-terminal sequences of all the phages suggests that stable 'panhandle' structures are unlikely to be formed via base pairing of both ends . However, thermodynamically more stable panhandle structures might be formed by displaced single-stranded DNA, although this requires rather large loops.

Gene, 1985, 35(1-2), 151 - 7
Genes for Bacillus megaterium small, acid-soluble spore proteins: nucleotide sequence of two genes and their expression during sporulation; Fliss ER et al.; The complete nucleotide (nt) sequence of two Bacillus megaterium genes coding for small, acid-soluble spore proteins (SASP), termed C-1 and C-2, has been determined . The nt sequences of the genes are greater than 98% identical in the coding regions, greater than 90% identical in approx . 180 bp and approx . 50 bp of upstream and downstream flanking sequences, respectively, and exhibit features conserved in related B . megaterium SASP genes . Northern blot analyses showed that the SASP-C-1 and/or C-2 genes are transcribed during sporulation in parallel with the related SASP-C and C-3 genes . The promoter regions of the SASP-C-1 and C-2 genes were localized, based on the sizes of their mRNAs and the positions of transcription termination sequences . The SASP-C-1 and C-2 genes' promoter regions exhibit significant homology with those for the SASP-C and C-3 genes.

Microbiol Immunol, 1985, 29(2), 119 - 26
Inhibition of cortex hydrolysis during spore germination by CdCl2; Nakatani Y et al.; When the spores of Bacillus megaterium QM B1551 (ATCC 12872) were incubated with 5 mM CdCl2 at 30 C, they underwent the early germination events, such as loss of heat resistance and release of calcium dipicolinate, in the same way as when they were germinated by glucose + KNO3 . However, germination by CdCl2 caused no increase in the reducing groups in the cortex and no excretion of glucosamine-containing materials due to the hydrolysis of the cortex peptidoglycan . Addition of CdCl2 at any time during germination by glucose + KNO3 inhibited the release of glucosamine-containing materials from the spores, whereas removal of cadmium from the CdCl2-germinated spores by treatment with cysteine restored the hydrolysis of peptidoglycan . These results suggested that CdCl2 caused the early events of spore germination but prevented the spores from undergoing the events following germination by inhibiting the enzymatic lysis of the cortex peptidoglycan . The conclusion from the study is that cortex degradation is not always required for the initiation of germination.

Dev Comp Immunol, 1985 Winter, 9(1), 11 - 20
Protein analysis of earthworm coelomic fluid . III . Isolation and characterization of several bacteriostatic molecules from Eisenia fetida andrei; Vaillier J et al.; A bacteriostatic activity in Eisenia fetida andrei cell free coelomic fluid is described . This activity is detected by growth inhibition of a bacteria Bacillus megaterium . Gel filtration analysis revealed eleven coelomic fluid protein fractions designated A, B,..J . Antibacterial activity was mainly found within fractions B and C . Chromatofocusing resolved fractions B-C into five different peaks named alpha BC, beta BC,.. . epsilon BC . Antibacterial activity appeared mediated by three different proteins characterized by their molecular weights (20,000, 40,000 and 45,000) and their isoelectric points (4.9, 5.75 and 6.0) . These bacteriostatic proteins possess either hemolysis or hemagglutination activities . The polymorphic aspect of this humoral antibacterial defense is discussed.

Microbiol Immunol, 1985, 29(1), 21 - 37
Properties and origin of filamentous appendages on spores of Bacillus cereus; Kozuka S et al.; Some physical, chemical, and immunological properties of filamentous appendages and the exosporium on the spores of Bacillus cereus were examined for the purpose of elucidating the origin of filamentous appendages . The main components of both filamentous appendages and the exosporium were protein and their amino acid compositions were similar in point of a high content of glycine, alanine, threonine, valine, and acidic amino acids and a low content of basic and sulphur-containing amino acids . Treatment with 1 N NaOH at 50 C solubilized the isolated appendages completely and the isolated exosporia partially . In both preparations the solubilized proteins consisted of highly acidic monomeric subunits with molecular weights between 2,000 and 5,000 . Treatment of the spores with 2% 2-mercaptoethanol at 37 C resulted in the isolation of long filamentous appendages without segmentation . When the spores were treated with 10% 2-mercaptoethanol, there was partial destruction of the exosporium as well as detachment of the filamentous appendages . There was a common antigenic component in the exosporium and the tips of the filamentous appendages . Five strains of B . cereus having a common appendage antigen also had a common exosporium antigen, whereas six other strains had neither a common appendage antigen nor a common exosporium antigen . From these facts it was concluded that the filamentous appendages arose from the exosporium.

Comput Appl Biosci, 1985, 1(1), 23 - 7
Microcomputer assisted identification of Bacillus species; Bryant TN et al.; A microcomputer based system for the identification of unknown isolates of Bacillus species is described . The identification matrix includes 78 test probabilities for 38 recognised species and other groups in the genus Bacillus and it is based on the work of Logan and Berkeley (1984) . Morphological characters together with the results of tests using API 20E and API 50CHB, read after 24 and 48 h incubation, are used to obtain a probabilistic identification of an unknown aerobic endospore forming rod . Any differences between the observed and expected results for any identified organism are listed . Identification can be attempted on the basis of a limited set of test results, although this is rarely if ever done with this largely API based system, and if the unknown cannot be successfully identified then a set of additional tests can be selected which should permit identification . The computer system can store and recall test results entered for any isolate . This feature allows the accumulation of data on isolates which could be used to update the identification matrix in future taxonomic studies.

Zentralbl Allg Pathol, 1985, 130(4), 299 - 306
{Cohnheim's inflammation doctrine and the current debate}; Doerr W; In a historical view the development of the concept of inflammation and the assessment of its importance are considered in the light of the situation in the field of pathologic anatomy in the last century dominated as it was by the work of Cruveilhier, Rokitansky and Virchow . It is to Cohnheim that we owe the realization that inflammatory cells originate in the circulation . He attributed this to an alteration of the vessel wall . Years before the tuberculosis bacillus was discovered, Cohnheim recognized the disease as a single entity . He also distinguished "tumours" of infectious origin from true neoplasms . Observations made by Cohnheim continue to form part of the contemporary body of knowledge, for example the rejection of an acellular origin for inflammatory cells . Cohnheim's work provides the basis for current notions of allergy, granuloma formation and the pathologic changes associated with tuberculosis . Similarly our pathobiochemical views of inflammation are ultimately based on the appreciation of changes in microcirculation and capillary permeability.

Immunol Lett, 1985, 11(3-4), 189 - 94
T-dependent production and activation of mononuclear phagocytes during murine BCG infection; Milon G et al.; Mice infected with a high dose of viable Bacillus Calmette Guerin (BCG) intravenously offer an interesting model to study regulatory functions of T cells on hemopoiesis . The proposition that T lymphocytes may play such a regulatory role was tested in nu/nu and two genetically different strains of mice: while the hemopoiesis of C3H/He mice remained unchanged during BCG injection, that of infected C57BL/6 mice was rapidly and transiently modified towards increased production of phagocytes at the expense of the erythroid lineage . The number of BCG-specific T cells present in C57BL/6 bone marrow was 50-100 higher than that determined in C3H/He mice . Moreover, between day 0 and 5 of infection the majority of BCG-specific T cells in C57BL/6 animals were of the L3T4+ Lyt2- surface phenotype . An attempt was made to identify the nature of the T cell product(s) able to activate young bone marrow-derived macrophages to render them non-permissive to growth of BCG.

Med Pr, 1985, 36(2), 123 - 30
{Ototoxic factors requiring consideration in the diagnosis of occupational hearing loss}; Grzesik J et al.; In the practice of diagnosing occupational deafness resulting from noise effects of factors determining workers' hearing, such as living conditions, working conditions, nutritional and other habits, diseases and their therapy, are often neglected . Discussed in the paper are the significance and ototoxic effects of such factors as: aminoglycoside antibiotics, diuretics, salicylic acid derivatives, fenacetin, quinine, fluorine compounds, cytotoxic drugs, chemical compounds other than drugs (carbon monoxide, carbon disulphide, lead, organic solvents), ethyl alcohol, diseases (abdominal typhus, bacillary dysentery, diphtheria, brucellosis, epidemic parotiditis, poliomyelitis, rubella, aural shingles, syphilis, diabetes mellitus, chronic renopathies, hypothyroidism, serologic conflict, pigmentary retinitis) . Exposure to intense noise is more and more frequently juxtaposed with the impact of the mentioned factors . If industrial physicians get aware of this association the prevention of deafness and reliability of treatment may be largely promoted.

J Neurosurg Sci, 1985 Jan-Mar, 29(1), 19 - 24
Chemotherapy plus immunotherapy for patients with primary and metastatic brain tumors; Knerich R et al.; The authors report two clinical trials concerning chemotherapy and immunotherapy combined in the treatment of primary and metastatic brain tumors . In the first study bacillus Calmette-Guerin (BCG) and in the second Levamisole (LMS) were utilized as immunostimulating agents . Chemotherapy was performed with BCNU or CCNU in association with surgery and/or radiotherapy . The immunological response was obtained but immunotherapy failed to demonstrate any significant effect on survival.

Oncology, 1985, 42(5), 275 - 81
Effect of administration of BCG, levamisole and irradiated leukemic cells on immune status and remission status in chronic myelogenous leukemia; Advani SH et al.; The immunomodulating effects of bacillus Calmette-Guerin (BCG) and levamisole with and without irradiated autologous leukemic cells were tested in patients with chronic myeloid leukemia (CML) after control of white cell count with busulfan . The response has been evaluated with respect to remission period and in vivo and in vitro immunological studies comprising delayed cutaneous hypersensitivity response to recall antigens and dinitrochlorobenzene, T cell and its subsets percentages, T-cell response to phytohemagglutinin, and to leukemia cell extracts by blastogenesis and leukocyte migration inhibition . Patients receiving BCG or levamisole did show marginal prolongation of remission, however immune parameters failed to show any improvement . In contrast, improvements in specific and nonspecific immune parameters were observed in patients receiving BCG or levamisole along with irradiated leukemic cells, however, concomitant clinical benefit was not obtained . Development of a better immunotherapeutic approach appears essential for immunomodulation in CML.

Respiration, 1985, 48(1), 29 - 36
Histological changes in rat bronchus-associated lymphoid tissue after administration of five different antigens; van der Brugge-Gamelkoorn GJ et al.; Histological changes in bronchus-associated lymphoid tissue (BALT) following single intratracheal administration of five different antigens were studied in the rat . After administration of T-dependent antigens (i.e., horseradish peroxidase, bovine serum albumin, and Bacillus Calmette Guerin) only minor changes in BALT in the rat occurred . Intratracheal administration of a T-independent antigen (lipopolysaccharide) and a partly T-independent antigen (paratyphoid vaccine) resulted after 1 week in an increase in cytoplasmic IgM-containing (cIgM) blast cells and plasma cells; these appeared to enter via the high endothelial venules (HEV) . After 6 weeks, germinal centers were seen.

Z Parasitenkd, 1985, 71(2), 249 - 57
Effects of macrophage activity on the antibody-dependent cytotoxicity against Trichinella spiralis newborn larvae: an in vitro cytotoxicity and ultrastructural study; Perrudet-Badoux A et al.; The effect of the activity of macrophages on the antibody-dependent cytotoxicity against Trichinella spiralis newborn larvae was studied in vitro . Macrophages present in peritoneal exudates from mice genetically selected for high and low antibody production (HL and LL, respectively) showed an inverse cytotoxic effect . Cells from HL mice were ineffective, whereas cells from LL mice had a very high killing capacity . Ultrastructural studies of cells after incubations of up to 36 h supported these observations . Furthermore, peritoneal macrophages from congenitally athymic (nu/nu) mice showed a higher killing potential than cells from thymus-bearing littermates (+/nu) mice . The activity of the latter cells could be increased by in vitro pretreatment of the mice with Calmette-Guerin bacillus, a well-known macrophage stimulating agent . The results indicate that macrophages, although not the only effector cells, may play an important role in the defence against T . spiralis newborn larvae.

Ther Drug Monit, 1985, 7(1), 12 - 25
An overview of amikacin; Ristuccia AM et al.; Amikacin, a semisynthetic analog of kanamycin, is very active against most gram-negative bacteria including gentamicin- and tobramycin-resistant strains . The effectiveness of amikacin in the treatment of serious gram-negative bacillary infections is well documented . Due to its resistance to inactivating enzymes, it is the aminoglycoside of choice for the treatment of known or suspected serious gram-negative infections caused by organisms resistant to gentamicin or tobramycin . Amikacin should be part of an empiric antibiotic regimen for the therapy of suspected sepsis in febrile, leukopenic immunocompromised hosts since it exhibits enhanced activity against the organisms most frequently encountered in this patient population . High response rates have been reported with the use of amikacin combined with beta-lactam antibiotics in immunocompromised or granulocytopenic patients . It exhibits impressive in vitro synergy against aminoglycoside-sensitive and -resistant organisms when used in combination with the new acylureidopenicillins and third-generation cephalosporins . Amikacin has the advantage of being the aminoglycoside least inactivated by the semisynthetic penicillins . Amikacin achieves high and predictable serum concentrations and has a favorable therapeutic index . Its potential for nephrotoxicity and ototoxicity is not significantly different than that encountered with gentamicin or tobramycin . Amikacin appears to be the preferred aminoglycoside for use at the present time because of its activity against gentamicin- and tobramycin-resistant organisms, its low resistance potential, its relative low degree of inactivation by the semisynthetic penicillins, and its superior pharmacokinetic profile.

Appl Environ Microbiol, 1985 Jan, 49(1), 137 - 42
Autoradiographic method to screen for soil microorganisms which accumulate zinc; Zamani B et al.; An autoradiographic method was developed to screen for and isolate soil microorganisms which accumulate zinc (Zn) . Diluted soil samples (Rubicon fine sand, Entic Haplorthods {pH 5.9}) were plated on soil extract-glucose agar containing radioactive 65Zn . After 7 days of incubation, individual colonies which accumulated sufficient 65Zn could be detected by autoradiography . These colonies were isolated and confirmed as Zn accumulators in pure culture by using the autoradiographic plate technique . Most Zn accumulators were filamentous fungi, identified as Penicillium janthinellum, Aspergillus fumigatus, and Paecilomyces sp . Isolates of Penicillium janthinellum were the most common Zn accumulators . The most abundant Zn-accumulating bacteria were Bacillus spp . The validity of the autoradiographic plate technique to differentiate soil microbes which accumulate Zn was examined independently by energy dispersive X-ray analysis in a scanning electron microscope . This method confirmed that fungal isolates which gave positive autoradiographic responses in the plate assay bioaccumulated more Zn in their biomass than fungal isolates from the same soil sample which gave negative autoradiographic responses . Thus, this technique can be applied to specifically screen for and isolate microbes from the environment which bioaccumulate Zn.

Invest Ophthalmol Vis Sci, 1985 Jan, 26(1), 15 - 22
Removal of viable sheets of conjunctival epithelium with dispase II; Geggel HS et al.; A technique (based on Gipson and Grill, Invest Ophthalmol Vis Sci 23:269, 1982) has been developed to obtain pure, viable, intact sheets of rabbit conjunctival epithelium free of the underlying basement membrane . After preparing a full thickness eye wall resection, Dispase, grade II (neutral protease-Bacillus polymyxa), 1.2 Units/ml MEM is injected intrasclerally . The conjunctiva and sclera are pinned in agar and incubated in the dispase with MEM for 1 hour . A 2 X 3 mm sheet of conjunctival epithelium can be dissected bluntly . Light microscopy shows a two- to three-layered epithelium with many goblet cells . Transmission electron microscopy reveals blebbing at the freed basal epithelial cell membrane, intact desmosomes, and intact goblet cells . The conjunctival sheets were cultured on epithelial-scraped corneal stromal carriers in vitro . Numerous goblet cells were present up to 12 hours on 4-mm carriers and 24 hours on 1-mm carriers . With this technique, pure populations of conjunctival epithelium can be isolated for further characterization and tissue culture.

Gene, 1985, 40(2-3), 311 - 6
The complete sequence of Bacillus phage phi 29 gene 16: a protein required for the genome encapsidation reaction; Garvey KJ et al.; We have sequenced the region of the Bacillus phage phi 29 genome that encodes gene 16, the gene product of which catalyzes the in vivo and in vitro genome-encapsidation reaction . The identity of the coding frame was confirmed by sequencing a sus mutant, sus16(300) . It is concluded that gene 16 encodes a 39-kDal protein and is comprised of 331 amino acids . Only 30 bp separates gene 16 from the last open reading frame of the right early region . Analysis of potential secondary structures in this region suggests that the same sequences may be involved in the termination of both the late and early transcripts.

Rev Argent Microbiol, 1985, 17(3), 157 - 63
{Infection of a cerebrospinal fluid shunt system by Bacillus circulans and Bacillus larvae}; Roncoroni A et al.; A two episodes case of CSF ventriculo-atrial shunt infection due to B . circulans and B . larvae is presented . B . circulans was first isolated from 4 blood cultures and CSF (shunt valve tap) . The patient showed a brain damage syndrome reversible with antibiotic treatment . Lethal toxin production was demonstrated for the B . circulans strain in a mouse model . This strain was found to be a variant of Gordon's description as it produced urease and was tolerant to 7% NaCl . The patient recovered after cefotaxime, cotrimoxazole and rifampicin treatment . A second infection due to B . larvae was detected two months later . The shunt system was removed due to obstruction and a scanning electron microscopy study was performed . Confluent masses of white blood cells and rods were observed on the inner surface of the catheter . As for as we know, this is the first case of human infection due to B . larvae.

Ciba Found Symp, 1985, 111, 204 - 18
Redesigning enzymes by site-directed mutagenesis; Fersht AR et al.; The systematic alteration of protein structure has now become possible with genetic engineering . Recent developments in techniques for the chemical synthesis of DNA fragments and in recombinant DNA technology have enabled the facile modification of proteins by highly specific mutagenesis of their genes . Enzymes with novel properties may be produced in large quantities from the mutant genes . Kinetic analysis of the mutant enzymes can be combined with high-resolution structural data from protein X-ray crystallography to provide direct measurements on the relationships between structure and function . In particular, the strength and nature of enzyme-substrate interactions and their roles in catalysis and specificity may be studied . The tyrosyl-tRNA synthetase from Bacillus stearothermophilus is being systematically analysed by site-directed mutagenesis . A fine-structure analysis is revealing the subtle roles of hydrogen bonding in catalysis and specificity . Modification of the residues that hydrogen-bond with ATP and tyrosine shows how the energetics must be analysed in terms of an exchange reaction with solvent water . Based on this idea, and structural data, an enzyme of vastly improved enzyme-substrate affinity has been engineered . There thus appear to be real prospects of engineering proteins of new specificities, activities and structural properties . Direct information is also being gathered on the nature of enzyme catalysis . For example, the catalysis of formation of Tyr-AMP from Tyr and ATP does not appear to use the classical mechanisms of acid-base or covalent catalysis . Instead, there just appears to be a binding site that stabilizes the high-energy pentacoordinate intermediate in the reaction.

Angiology, 1985 Jan, 36(1), 64 - 6
Infectious endocarditis caused by Actinobacillus actinomycetemcomitans; Gould L et al.; Actinobacillus actinomycetemcomitans is a very uncommon cause of infectious endocarditis . The organism was first described in 1912 . Thjotta and Sydnes reported its isolation in pure culture from a long standing abscess which had developed after tooth extraction . Subsequently this organism was found to be part of the normal flora, and the organism was defined as a slow growing, fastidious gram negative bacillus . Carbon dioxide is essential for the growth of A . actinomycetemcomitans . Approximately 50 cases of endocarditis due to A . actinomycetemcomitans have been reported since the first case reported in 1964 . The purpose of this report is to document a case of endocarditis due to A . actinomycetemcomitans and to stress the value of the echocardiogram in the assessment of patients with endocarditis.

Indian J Lepr, 1985 Jan-Mar, 57(1), 27 - 36
Influence of levamisole on lymphocytes and M . leprae in mice; Ganguly NK et al.; Normal uninfected (N) and M . leprae infected mice (NI) were given levamisole in the dose of 2.5 mg/kg body weight . The animals were observed over a period of nine months for bacillary load, T and B cell counts and blast transformation with PHA . Significant increase in B cell counts was observed in the levamisole treated normal (NL) compared to normal control (N) group . T cell counts and blast transformation, however, remained unaffected . However, T-cell counts and blast transformation improved significantly in the infected and levamisole treated (NIL) as compared to the infected group (NI) not given levamisole . Bacillary loads remained unaltered in both, the infected (NI) and levamisole treated (NIL) group.

Indian J Lepr, 1985 Jan-Mar, 57(1), 159 - 63
Multi-drug therapy for multi-bacillary cases in Wardha district, Maharashtra, India; Rao MS et al.; 2786 infectious patients of Wardha district are now receiving multi-drug therapy . 67% of those who have received 2 years' treatment have become bacteriologically negative . Side-effects due to drugs have been comparatively few . In the majority of patients who had such side-effects, they have been mild.

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