Microbiologica, 1992 Apr, 15(2), 157 - 65 Phenoloxidase production and vanillic acid metabolism by Zygomycetes; Seigle-Murandi F et al.; The ability of 23 strains of Zygomycetes to produce extracellular phenoloxidases was examined on solid media by using 10 different reagents . The results varied depending on the reagent and indicated that most of the strains were devoid of phenoloxidase activity . The production of inducible phenoloxidases was demonstrated by the Bavendamm reaction . The study of the biotransformation of vanillic acid in synthetic medium indicated that the reaction most often obtained was the reduction of vanillic acid to vanillyl alcohol . Helicostylum piriforme and Rhizopus microsporus var . chinensis completely metabolized vanillic acid while good transformation was obtained with Absidia spinosa, Cunninghamella bainieri, Mucor bacilliformis, Mucor plumbeus, Rhizopus arrhizus, Rhizopus stolonifer, Syncephalastrum racemosum and Zygorhynchus moelleri . Other strains did not degrade or poorly degraded vanillic acid . Decarboxylation and demethoxylation of this compound was independent of the production of phenoloxidases as in the case of white-rot fungi . Other enzymatic systems might be implicated in this phenomenon. Kekkaku, 1992 Apr, 67(4), 319 - 29 {Surveillance of tuberculosis epidemics in Aichi Prefecture}; Ikari A; A survey was done on the conditions of the tuberculosis surveillance system used for extraordinary examination in high tuberculosis prevalence areas in Aichi Prefecture during the period from 1981 to 1989 . The results were as follows: 1) A total of 25 health centers provided information to 456 cases in specific groups known as "danger groups" and 227 cases within a furniture company . Information was most frequently provided when the index cases had been diagnosed bacillary positive . 2) Extraordinary examinations were performed on 222 cases for nine years, one case a year per each health center . The frequency of extraordinary examination was never high . A total of five epidemic cases (2.3%) and 30 microepidemic cases (13.5%) were discovered . 3) Coordination among health centers to make examinations was required in 81 cases (36.5%) . 4) In index cases in which extraordinary examinations were performed, 43 preventable cases (19.4%) were found . It is therefore important to thoroughly enforce family-contact examinations . In order to complete extraordinary examinations, the importance of not only the systematic collection of information and the action of examination about specific groups but also the establishment of a surveillance system for extraordinary examinations is vital. Plant Mol Biol, 1992 Apr, 18(6), 1133 - 9 Direct screening for high-level expression of an introduced alpha-amylase gene in plants; Pen J et al.; A method is described for obtaining transgenic plants with a high level of expression of the introduced gene . Tobacco protoplasts were transformed with an expression construct containing a translational fusion between mature alpha-amylase from Bacillus licheniformis and the signal peptide of the tobacco PR-S protein . A total number of 5200 transformed protoplasts was cultured to microcalli and screened for alpha-amylase expression by incubation on media containing starch followed by staining with iodine . The calli were divided into four classes, based on the resulting halo sizes on the plates . The halo sizes were found to correlate with the expression levels in transgenic plants regenerated from the calli . The expression levels varied between 0 and 0.5% of soluble leaf protein in the regenerated transgenic plants . Wider implications of this method are discussed. Wei Sheng Wu Xue Bao, 1992 Apr, 32(2), 99 - 104 {Screening of strain producing extracellular penicillin acylase}; Wang Z et al.; Ninety-eight strains having extracellular penicillin acylase activity were derived from soil samples by colour-developing method . 10 strains of them possess higher activity of penicillin acylase . All of those are found to be Bacillus megaterium . The optimum condition of enzyme production was investigated with the strain No . 46 which is from No . 247 by single colony isolation . The productivity of penicillin acylase in the optimum condition have been enhanced 2.5 times more than that in the screening condition . The mutant strain, Bacillus megaterium UL-81, which penicillin acylase activity reached the level of 723u/100ml of broth was obtained from No . 46 by treatment with physical and chemical factors . The penicillin acylase activity of UL-81 can reach 820u/100 ml in 500L fermentor . The mutant strain differed from parent strain in the morphology of colony, the size of cells, the effect of concentration and the addition time of phenylacetic acid on the production of penicillin acylase. Hautarzt, 1992 Apr, 43(4), 184 - 9 {Immunology of leprosy . Review of data from the 1980-1990 time period}; Castells Rodellas A et al.; The classification used for leprosy so far distinguishes between two polar forms (pauci-bacillary and multi-bacillary), with different influences on the immune cells . The identification of specific antigens of the germ surface with monoclonal antibodies allows finer differentiation in classification, which now extends to transitional forms of clinical relevance . The current status of knowledge about immunological phenomena triggered by Mycobacterium leprae is complex, and controversy is rife at present . The bacillus is comparable to other mycobacteria in antigenicity . Owing to the complicated composition of the cellular envelope, few epitopes are accessible for immunological reactions . Investigation on the behaviour of T-lymphocytes in combination with analysis of the bacterial surface structures led to the development of a new pathogenetic concept . In spite of different immunological and inflammatory reactions in patients (reflecting the individual pathology), the discrimination between a partial and a total immunological defect is made . So far, no histocompatibility-antigen type has been found that allows recognition of the predisposition to infection . Immunity against the bacteria is achieved after BCG in some cases . All this suggests that the development of a vaccine via biotechnical synthesis of idiotypes and anti-idiotypes may be possible. Immunobiology, 1992 Apr, 184(4-5), 336 - 47 Effect of Bacillus Calmette-Guérin inoculation on numbers of dendritic cells in bronchoalveolar lavages of rats; Havenith CE et al.; Dendritic cells (DC) are present in all lymphoid tissues and are widely distributed in the airway epithelium and lung parenchyma . In this study DC were morphologically and cytochemically identified in normal rat bronchoalveolar lavage (BAL), although in very low percentages . Furthermore, the total population as well as different Percoll density fractions demonstrated poor antigen-presenting capacity and even suppressed antigen-specific stimulation by rat splenic DC . In contrast, when an inflammatory response was induced by intratracheal inoculation of Bacillus Calmette-Guerin (BCG), an increase of Ia-positive cells, containing high percentages of monocytes and DC (MDC) was found . In BAL, DC increased about 25 times within 48 h after BCG inoculation . These BCG-induced BAL cells as well as the different density fractions showed a high antigen-presenting capacity at low concentrations . However, at higher concentrations they were suppressive, except for the highest density fraction which lacked alveolar macrophages (AM) . These results indicate that the increased numbers of Ia-positive MDC during an inflammatory reaction are very likely responsible for antigen presentation in vitro . In contrast, AM suppress the antigen-specific T cell proliferation in a concentration dependent manner. Mol Microbiol, 1992 Apr, 6(8), 1013 - 23 A bifunctional xylanase encoded by the xynA gene of the rumen cellulolytic bacterium Ruminococcus flavefaciens 17 comprises two dissimilar domains linked by an asparagine/glutamine-rich sequence; Zhang JX et al.; The nucleotide sequence of the xynA gene of Ruminococcus flavefaciens 17 was determined and found to consist of a 2862bp open reading frame beginning with a TTG start codon . The predicted product, XYLA, consisted of distinct amino-terminal (A) and carboxy terminal (C) domains (248 amino acids, including a putative signal sequence, and 332 amino acids, respectively) linked by a repetitive sequence (B, 374 amino acids) extraordinarily rich in asparagine (45%) and glutamine (26%) residues . Domains A and C were shown to be capable of expressing xylanase activity independently of each other when suitably truncated derivatives of the xynA coding region were expressed as lacZ fusions . The activities associated with the two domains were shown to differ with respect to the average size of hydrolysis products formed from oat-spelt xylan, with domain C releasing relatively more xylose and domain A more xylo-oligosaccharides . The amino acid sequence of domain A of XYLA closely resembled that of the Bacillus pumilus xynA enzyme (45% identical residues) . On the other hand domain C showed significant similarity (33% to 40% identical residues) to a different group of bacterial xylanases and exoglucanases exemplified by the Caldocellum saccharolyticum xynA and celB products . The xynA product is, therefore, a bifunctional enzyme having two dissimilar catalytic domains capable of acting on xylan. Proteins, 1992 Apr, 12(4), 331 - 8 Substrate-enzyme interactions and catalytic mechanism in phospholipase C: a molecular modeling study using the GRID program; Byberg JR et al.; Based on the high-resolution X-ray crystallographic structure of phospholipase C from Bacillus cereus, the orientation of the phosphatidylcholine substrate in the active site of the enzyme is proposed . The proposal is based on extensive calculations using the GRID program and molecular mechanics geometry relaxations . The substrate model has been constructed by successively placing phosphate, choline and diacylglycerol moieties in the positions indicated from GRID calculations . On the basis of the resulting orientation of a complete phosphatidylcholine molecule, we propose a mechanism for the hydrolysis of the substrate. Jikken Dobutsu, 1992 Apr, 41(2), 231 - 4 Propagation of CAR bacillus in artificial media; Shoji Y et al.; CAR bacillus propagated successfully in an artificial medium, and the number of CAR bacillus was about 30 times the original number after 8 days of cultivation . The medium consisted of Eagle's minimum essential medium supplemented with 10% fetal calf serum and 20% hamster tracheal organ culture soup . By intranasal inoculation to mice, two strains of the CAR bacillus passaged 5 and 6 times in this artificial medium produced the same lung lesions as natural CAR bacillus infection. Jikken Dobutsu, 1992 Apr, 41(2), 181 - 7 {Efficacy of ozone fumigation for laboratory animal sanitation}; Pan TM et al.; Ozone resistance of spores of 2 strains of Bacillus isolated from laboratory animals was examined . Each of 0.02 ml of phosphate-buffered saline at pH 7.0 containing 10(6) Bacillus spores was dropped onto sterilized filter strips, wood chip bedding, pellets of diet, cloth pieces and stainless steel plates . After drying at room temperature, the test materials were exposed to ozone gas of different concentrations at 90% RH . Exposure to 200ppm ozone for 6 hours was sufficient to kill spores in filter strips, but a little higher concentration or a little longer period of ozone fumigation was necessary for sterilization of wood chips, cotton cloth pieces and steel plates . The present results indicated that 600ppm ozone fumigation for 6 hours might be effective for routine sterilization of cages, wood bedding, working clothes and other materials used in laboratory animal facilities . However, exposure to ozone gas of 500 or 1,000 ppm for 6 hours or 200 ppm for 24 hours could not kill spores in pellets of diet, suggesting that dietary protein inhibited the bactericidal activity of ozone. Clin Infect Dis, 1992 Apr, 14(4), 945 - 6 Bacillus cereus endocarditis: report of a case and review; Steen MK et al.; Bacillus cereus is a ubiquitous organism that infrequently causes serious infections . We report a patient with B . cereus endocarditis involving a mechanical aortic valve . Data for 10 cases of B . cereus endocarditis reported in the literature are summarized . B . cereus is resistant to many commonly used antibiotics, a finding that has clinical significance for empirical antibiotic selection in patients with suspected endocarditis . Infection in patients with valvular heart disease in the few cases reported is associated with significant mortality and morbidity. West J Med, 1992 Apr, 156(4), 423 - 5 Pulmonary paragonimiasis in Southeast Asians living in the central San Joaquin Valley; Yee B et al.; We describe four cases of pulmonary paragonimiasis in Southeast Asians who emigrated to the central San Joaquin Valley of California . Physicians should be alerted to the possibility of this disease in Asian patients with hemoptysis and pulmonary infiltrates . Paragonimiasis can masquerade as pulmonary tuberculosis, especially in patients from areas that are endemic for both the parasite and the tubercle bacillus . The ease and safety with which this infection can be treated, in contrast to tuberculosis, reiterates the importance of diagnosing this lung fluke when it is present. Gene, 1992 Apr 1, 113(1), 83 - 8 Overproduction, purification and characterization of M.HinfI methyltransferase and its deletion mutant; Bassing CH et al.; We have used the polymerase chain reaction to alter transcriptional and translational signals surrounding the hinfIM gene {encoding M.HinfI methyltransferase (MTase)} so as to achieve overexpression in Escherichia coli . The PCR-generated hinfIM gene was subcloned in a high-expression vector under control of the hybrid trp-lac promoter . In addition, the positive retroregulator stem-loop sequence derived from the crystal protein-encoding gene of Bacillus thuringiensis was inserted downstream from hinfIM . Using a similar approach, we have also constructed overproducer clones of a deletion mutant of M.HinfI MTase that has 97 amino acids from the C terminus deleted . The plasmid from the mutant clones is fully protected from HinfI restriction endonuclease digestion . It appears that the functional properties (the recognition and catalytic functions) are encoded within this mutant gene . The overproducer clones yield the wild type (wt) and the mutant enzymes to about 10% of total cellular protein upon induction with 1 mM IPTG . The wt M.HinfI and the mutant MTase were purified to near electrophoretic homogeneity by phosphocellulose, DEAE and gel chromatography . Their monomer sizes by SDS/polyacrylamide-gel electrophoresis are 43 kDa and 31 kDa, respectively, in good agreement with that predicted from the nucleotide sequence . DNA methylation experiments with purified enzymes using single-strand and double-strand M13mp18 DNA substrates indicate that while wt enzyme methylates both forms of DNA substrates, the mutant enzyme appears to preferentially methylate ss DNA substrate. Urology, 1992 Apr, 39(4), 364 - 7 BCG-induced granulomatous prostatitis: a comparative ultrasound and pathologic study; Miyashita H et al.; We analyzed the ultrasound findings and the information obtained from serial whole-mount cross-section histologic examination of the prostates from 6 patients with bacillus Calmette-Guerin (BCG)-induced granulomatous prostatitis . Similarities to prostate cancer included a hypoechoic pattern on ultrasound, multifocality, and a propensity for a peripheral zone location . BCG-induced granulomatous prostatitis cannot be distinguished from cancer by ultrasonography . We suggest performing an ultrasound examination prior to starting BCG immunotherapy as a baseline for future reference to aid in the clinical diagnosis of BCG-induced granulomatous prostatitis. J Urol, 1992 Apr, 147(4), 1020 - 3 Bacillus Calmette-Guerin therapy for superficial bladder cancer: a 10-year followup; Herr HW et al.; We report the outcome of 61 patients with superficial bladder tumors who received bacillus Calmette-Guerin (BCG) therapy and were followed for at least 10 years (range 10 to 13 years) . A total of 19 patients (31%) remains free of tumor and progression, 17 (28%) had interval superficial recurrences but no progression and 25 (41%) had progression, first identified as muscle invasion in 12, prostatic involvement in 8 or metastasis in 5 . Most tumors recurred or progressed within the first 5 years . Of the 61 patients 33 (54%) are disease-free and with an intact bladder, 13 (21%) are alive after cystectomy, 2 (3%) died of other causes, 1 (2%) is alive with metastasis and 12 (20%) died of metastatic urothelial cancer (11 from the bladder or prostate and 1 from an upper tract tumor). Indian J Exp Biol, 1992 Apr, 30(4), 324 - 6 Mechanisms of Bacillus cereus enteropathy; Singh DK et al.; Three strains of Bacillus cereus isolated from sausages (Salami and Trekker, RANBAC, Ranchi) produced enterotoxin which caused vascular permeability in skin and haemorrhage in the ligated ileal loops of rabbits . Histopathological studies revealed haemorrhage and congestion in submucosa, mononuclear cell infiltration in lamina propria and submucosa and villous atrophy . Histochemical studies ruled out the effect on mitochondrial enzymes of intestinal epithelial cells . Purified enterotoxin given intradermally to rabbits caused severe necrotic reaction at the site of injection and death within 4 hr . Histopathological changes observed in liver included congestion of portal veins and sinusoids, vacuolar degeneration of hepatocytes, and hyperplasia of bile ducts . These suggested that B . cereus enterotoxin affected the capillaries of blood vessels locally and also systemically resulting into release of proteinaceous exudates and red blood cells. Anal Biochem, 1992 Apr, 202(1), 61 - 7 Enzymatic synthesis of p-nitrophenyl 4(5)-O-beta-D-galactosyl-alpha-maltopentaoside as a substrate for human alpha-amylases; Usui T et al.; Enzymatic modification at the nonreducing end D-glucosyl residue of p-nitrophenyl alpha-maltopentaoside was developed by using the transglycosylation of beta-D-galactosidase from Bacillus circulans . The enzyme regioselectively synthesized p-nitrophenyl 4(5)-O-beta-D-galactosyl-alpha-maltopentaoside (a yield of 12.0% based on the amount of p-nitrophenyl alpha-maltopentaoside added) on a preparative scale from lactose as a donor and p-nitrophenyl alpha-maltopentaoside as an acceptor . It revealed that the nonreducing end galactosyl group of p-nitrophenyl 4(5)-O-beta-D-galactosyl-alpha-maltopentaoside did not prohibit the action of human salivary and pancreatic alpha-amylases . This derivative was shown to be very suitable as a novel substrate for analytical use of human alpha-amylase assay in serum through a conjugated reaction involving glucoamylase and alpha-D-glucosidase. Biotechnology (N Y), 1992 Apr, 10(4), 418 - 21 Expansion of insecticidal host range of Bacillus thuringiensis by in vivo genetic recombination; Lereclus D et al.; We describe a novel approach for the insertion of an insecticidal toxin gene into a resident plasmid in Bacillus thuringiensis (Bt) . A gene encoding a coleopteran-specific toxin was cloned within a fragment of IS232 and inserted into a plasmid thermosensitive for replication in Bt . The plasmid was used to transform a Bt strain toxic to lepidoptera, and the transformants were then selected at non-permissive temperature for clones in which the vector had integrated into a copy of IS232 present on a resident plasmid . A second recombination event was selected such that the vector was eliminated and the newly introduced toxin gene was conserved . The resulting strain contained only DNA of Bt origin, and displayed insecticidal activity against both lepidoptera and coleoptera. Pneumoftiziologia, 1992 Apr-Sep, 41(2-3), 113 - 8 {AIDS and tuberculosis: the immunopathogenic processes}; Paunescu E; HIV infection is characterized by CD4+ lymphocyte depletion manifested through the loss of the immune response capacity . The resulting immunodeficit is expressed by the blocking of immune surveillance mechanisms and, thus, by the establishment of favourable conditions to the development of opportunistic infections and/or malignant processes . In tuberculosis, the immunodeficiency associated with HIV infection makes possible the evolution of a latent infection to a clinically manifest disease . Latent tuberculosis is characterized by the intracellular persistence of some metabolically inactive Tb bacillus forms which are incapable of multiplication . The conversion of these inactive into metabolically active forms capable of multiplication is usually neutralized by immunosurveillance mechanisms . The blocking of such mechanisms in case of CD4+ cell depletion will allow the multiplication of metabolically active Tb bacillus forms, and the development of a clinically manifest tuberculosis . CD4+ lymphocyte depletion is the result of facilitating antibodies and certain cytokines, and of some autoimmune processes which also affect the non-infected CD4+ cells . Therefore, it is necessary that Tb chemoprophylaxis in HIV infected subjects should also be addressed to the processes initiating the immune deficit, which include autoimmune mechanisms as well as those facilitating HIV infection. Can J Microbiol, 1992 Apr, 38(4), 354 - 7 Biochemical defects of outermost layer deficient mutants during sporulation of Bacillus megaterium; Takubo Y et al.; To determine the regulation of morphogenesis of the outermost layer, the thick layer outside the inner coat, of the Bacillus megaterium spore, we isolated 15 outermost layer deficient mutants of B . megaterium using transposon Tn917 . Three mutant strains lacked both synthesis of the 48-kDa outermost layer protein and induction of two initial enzymes for galactosamine-6-phosphate polymer synthesis, evidence that these biochemical events are regulated in the cascade system during morphogenesis of the outermost layer. Rev Argent Microbiol, 1992 Apr-Jun, 24(2), 67 - 72 Characterization of Bacillus larvae White, the causative agent of American foulbrood of honey-bees . First record of its occurrence in Argentina; Alippi AM; American foulbrood caused by Bacillus larvae White is recorded for the first time on brood combs of Argentinian hives . The identification of the causative agent was based on disease symptomatology, morphological characters, pathogenicity tests and physiological and biochemical reactions . Studies by scanning electron microscopy showed the occurrence of large flagellar bundles of Bacillus larvae strains growing in biphasic BL medium . An electron microscope survey of the surface configuration of bacterial spores was also made. J Chromatogr, 1992 Mar 27, 575(2), 183 - 96 Quantification of distinct molecular species of the 2-lyso metabolite of platelet-activating factor by gas chromatography-negative-ion chemical ionization mass spectrometry; Turk J et al.; The biological activity of platelet-activating factor (PAF) is comprised by a few molecular species of phosphatidylcholine which contain a fatty alcohol connected by an ether linkage to the sn-1 position of the glycerol backbone and an acetate ester at the sn-2 position . The various molecular species of PAF differ in chain length and degree of unsaturation in the fatty alcohol residue side-chain . PAF is rapidly hydrolyzed to lyso-PAF by an acetylhydrolase enzyme which is quite active in a number of cells that synthesize PAF . We describe a method for quantitation of lyso-PAF which involves conversion to its propionate derivative in the presence of an internal standard (deuterium-labelled PAF), digestion to the diglyceride with Bacillus cereus phospholipase C, conversion to the pentafluorobenzoate derivative and capillary column gas chromatographic-negative-ion methane chemical ionization mass spectrometric analysis . Distinct molecular species of lyso-PAF can be individually quantitated at levels of 1 ng or less . These methods are applied to the demonstration of lyso-PAF accumulation in renal tissue from transplanted allografts undergoing acute rejection, in renal tissue from kidneys subjected to cold storage and autotransplantation, and in intestinal mucosa subjected to warm ischemia and reperfusion. Biochim Biophys Acta, 1992 Mar 16, 1134(2), 89 - 95 Interaction of energized bacteria cells with particles of colloidal gold: peculiarities and kinetic model of the process; Ulberg ZR et al.; It is found that the cells of Bacillus cereus B-4368 at energized state can concentrate the colloidal gold particles on their surface . It is shown that the process depends on metabolic reactions proceeding on the plasma membrane . The inhibitory analysis permits to suppose that the metal concentration is due to the functioning of ATP-dependent generator of the transmembrane potential, apparently, of proton ATPase . Kinetic characteristics of the process show the presence of an intermediate state in the formation of biomineral aggregates . A kinetic model of the studied process is suggested which describes the experimental data well. Carbohydr Res, 1992 Mar 16, 226(1), 119 - 30 Enzymatic synthesis of low molecular weight amyloses with modified terminal groups; Niemann C et al.; Low molecular weight amyloses with modified terminal groups were synthesized by cyclomaltohexaose (alpha-cyclodextrin) transfer using (1----4)-alpha-D-glucan: 4-alpha-D-(1----4)- alpha-D-glucopyranosyltransferase (cyclising) (EC 2.4.1.19) from Bacillus macerans . 4-Nitrophenyl alpha-malto-oligosaccharides d.p . 2-7 served as acceptors, and cyclomaltohexaose served as the donor . The reaction was optimized to obtain a majority of species of definite chain lengths in a range of d.p . 10-20, depending upon the chain length of the acceptor . The course of the coupling reactions, as well as the action of the enzyme in disproportionation, cyclisation, and hydrolysis of the products, were observed by h.p.l.c . analysis of the oligomer distributions . Using a 15-fold molar excess of cyclomaltohexaose and 0.5 units enzyme per mumol of acceptor at pH 5.2, the chromatograms revealed that the products of the coupling reaction were predominant during the first reaction period . By incubating the acceptors with the enzyme, but without the donor, the mechanism of disproportionation was elucidated as a transfer of malto-oligosaccharyl residues dependent upon the substrate chain length . The minimum chain length required for a direct cyclisation reaction was d.p . 7 . The results were confirmed by separation and investigation of the products of hydrolysis and cyclisation, which were nonmodified alpha-malto-oligosaccharides and cyclomalto-oligosaccharides. FEMS Microbiol Lett, 1992 Mar 15, 70(3), 277 - 84 Physical characterisation and over-expression of the Bacillus caldotenax superoxide dismutase gene; Chambers SP et al.; The gene (sod) encoding Bacillus caldotenax (BC) Mn-superoxide dismutase (MnSOD) has been cloned in Escherichia coli and its entire nucleotide sequence determined . Within the coding region of the gene there were 21 nucleotide differences to the previously sequenced sod of Bacillus stearothermophilus (BS) . The predicted amino acid sequence of BCMnSOD had two amino acid dissimilarities to the BSMnSOD, containing Asp and Val at positions 13 and 188, respectively, compared to Glu and Ile at the respective equivalent positions of BSMnSOD . Recombinant BCMnSOD was shown to be functionally active in E . coli, both in vitro and in vivo, and was produced at levels representing over 40% of the cells' soluble protein by coupling sod transcription to the E . coli trp promoter . The sequenced region of DNA was also found to encompass part of a second open-reading frame, of unknown function, previously noted 3' to the B . stearothermophilus gene. Proc Natl Acad Sci U S A, 1992 Mar 15, 89(6), 2056 - 60 A positive selection vector for cloning high molecular weight DNA by the bacteriophage P1 system: improved cloning efficacy; Pierce JC et al.; The bacteriophage P1 cloning system can package and propagate DNA inserts that are up to 95 kilobases . Clones are maintained in Escherichia coli by a low-copy replicon in the P1 cloning vector and can be amplified by inducing a second replicon in the vector with isopropyl beta-D-thiogalactopyranoside . To overcome the necessity of screening clones for DNA inserts, we have developed a P1 vector with a positive selection system that is based on the properties of the sacB gene from Bacillus amyloliquefaciens . Expression of that gene kills E . coli cells that are grown in the presence of sucrose . In the new P1 vector (pAd10sacBII) sacB expression is regulated by a synthetic E . coli promoter that also contains a P1 C1 repressor binding site . A unique BamHI cloning site is located between the promoter and the sacB structural gene . Cloning DNA fragments into the BamHI site interrupts sacB expression and permits growth of plasmid-containing cells in the presence of sucrose . We have also bordered the BamHI site with unique rare-cutting restriction sites Not I, Sal I, and Sfi I and with T7 and Sp6 promoter sequences to facilitate characterization and analysis of P1 clones . We describe here the use of Not I digestion to size the cloned DNA fragments and RNA probes to identify the ends of those fragments . The positive selection P1 vector provides a 65- to 75-fold discrimination of P1 clones that contain inserts from those that do not . It therefore permits generation of genomic libraries that are much easier to use for gene isolation and genome mapping than are our previous libraries . Also, the new vector makes it feasible to generate P1 libraries from small amounts of genomic insert DNA, such as from sorted chromosomes. J Immunol, 1992 Mar 15, 148(6), 1835 - 40 Immunoreactivity of a 10-kDa antigen of Mycobacterium tuberculosis; Barnes PF et al.; Identification of Ag of Mycobacterium tuberculosis recognized by T cells is essential to understanding the pathogenesis of tuberculosis and mechanism(s) of resistance to infection . Previous studies evaluating the immunoreactivity of nitrocellulose transfers of M . tuberculosis Ag separated by SDS-PAGE indicated that a high proportion of M . tuberculosis-reactive T cell lines proliferate in response to a 10-kDa Ag . We therefore purified this Ag from M . tuberculosis culture filtrates and evaluated its immunoreactivity in patients with tuberculous infection . Proliferative responses of PBMC to the 10-kDa Ag were similar to those induced by whole M . tuberculosis and greater than those elicited by other proteins isolated from culture filtrate . Furthermore, in patients with tuberculous pleuritis, proliferative responses to the 10-kDa Ag were higher in pleural fluid mononuclear cells than in PBMC, indicating that T cell reactivity to this Ag is enhanced at the site of disease . The first 15 amino acids of the 10-kDa Ag were identical to those defined previously for Bacillus Calmette-Guerin-a (BCG-a), and a T cell clone recognized the 10-kDa Ag and a peptide of BCG-a, indicating that the 10-kDa Ag corresponds to BCG-a . This Ag elicited IFN-gamma production by pleural fluid mononuclear cells and by PBMC from healthy tuberculin reactors, suggesting that the 10-kDa Ag can enhance macrophage activation and resistance to mycobacterial infection . Our findings indicate that the 10-kDa Ag of M . tuberculosis is highly immunoreactive and should be evaluated for its capacity to elicit protective immunity. Lancet, 1992 Mar 14, 339(8794), 636 - 9 Efficacy of BCG vaccine against leprosy and tuberculosis in northern Malawi; Ponnighaus JM et al.; Protection afforded by BCG (bacillus Calmette-Guerin) vaccines against tuberculosis and leprosy varies widely between different populations . In the only controlled trial which assessed protective efficacy of BCG (Danish and Pasteur strains) against both diseases, there was slightly more protection against leprosy than against tuberculosis . We have studied the protective efficacy of BCG (Glaxo, freeze dried) vaccine against these two diseases in Karonga District, northern Malawi . BCG vaccination was introduced into this population in 1974 . Prior information about BCG scar status was available for 83,455 individuals followed up between 1979 and 1989 . 414 new cases of leprosy and 180 new cases of tuberculosis were found in this population over that period . Protection was estimated at 50% or greater against leprosy, and there was no evidence for lower protection against multibacillary (84%; 95% confidence interval 26% to 97%) than against paucibacillary (51%; 30% to 66%) disease . There was no statistically significant protection by BCG against tuberculosis in this population . These findings add to the evidence that BCG vaccines afford greater protection against leprosy than against tuberculosis. J Urol, 1992 Mar, 147(3), 743 - 6 Effects of photodynamic therapy in combination with intravesical drugs in a murine bladder tumor model; Cho YH et al.; This study was designed to evaluate the interaction of photodynamic therapy (PDT) and intravesical drugs (thiotepa, adriamycin, mitomycin C and BCG) in a murine transitional cell carcinoma (MBT-2) model . C3H/He mice with implanted MBT-2 flank tumors were treated with either thiotepa (TT), adriamycin (ADM), mitomycin C, Bacillus Calmette-Guerin (BCG), photodynamic therapy (PDT) or a combination of the drug and PDT . The MBT-2 tumor showed sensitivity to adriamycin, MMC or PDT compared to control . PDT combined with either adriamycin, MMC or BCG, produced a greater retardation in the growth of the MBT-2 tumor than monotherapy with adriamycin, MMC, BCG or PDT . PDT combined with the anticancer agents currently used in intravesical therapy for bladder cancer is well tolerated . The combination of PDT and intravesical drugs may enhance the tumoricidal effect of either treatment used alone. J Biol Chem, 1992 Mar 5, 267(7), 4897 - 903 Membrane-bound aminopeptidase P from bovine lung . Its purification, properties, and degradation of bradykinin; Simmons WH et al.; The membrane-bound form of aminopeptidase P (aminoacylprolyl-peptide hydrolase) (EC 3.4.11.9) was purified to apparent homogeneity from bovine lung microsomes . The enzyme was solubilized using phosphatidylinositol-specific phospholipase C (Bacillus thuringiensis), indicating that bovine lung amino-peptidase P is attached to membranes via a glycosylphosphatidylinositol anchor . The enzyme was purified 1900-fold with a yield of 25% by chromatography on decyl-agarose, omega-aminodecyl-agarose, a second decylagarose column, DEAE-Sephacel, and an ultrafiltration step . Native gradient polyacrylamide gel electrophoresis revealed a single stained protein band whose position in the gel corresponded to cleavage of the Arg1-Pro2 bond of bradykinin . The Mr was 360,000 by gel permeation chromatography and 95,000 by reducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis . The substrate specificity of aminopeptidase P was determined using approximately 50 peptides with proline in the second position . The enzyme could hydrolyze lower NH2-terminal homologs of bradykinin, including Arg-Pro-Pro, which was used as the routine substrate in a rapid fluorescence assay performed in the absence of added Mn2+ . Some peptides having NH2-terminal amino acids other than arginine were also cleaved . Aminopeptidase P appeared to favor peptides that had 2 proline residues or proline analogs in positions 2 and 3 of the substrate . In general, tripeptides having a single proline residue in position 2 were poor substrates . Aminopeptidase P was inhibited by a series of peptides, 3-8 residues long, having an NH2-terminal Pro-Pro sequence . The enzyme was also inhibited by metal-chelating agents, 2-mercaptoethanol (4 mM), p-chloromercuribenzenesulfonic acid, and NaCl at concentrations greater than or equal to 0.25 M . The purified enzyme had a pH optimum of 6.5-7.0 and was most stable in the basic pH range . A role for membrane-bound aminopeptidase P in the pulmonary inactivation of circulating bradykinin is proposed. Mycopathologia, 1992 Mar, 117(3), 133 - 8 Localized cutaneous nocardiosis in Japan . A new case; Hiruma M et al.; We report a case of localized cutaneous nocardiosis due to Nocardia asteroides, appearing on the anterior surface of the right forearm of a 29-year-old woman . Examination of the skin revealed a firm subcutaneous nodule measuring 32 x 20 mm and accompanied with a small ulcer . The histology of the lesion showed granulomatous changes interspersed with large numbers of polymorphonuclear leukocytes . On Gram staining, a few bacillary bodies were found scattered through the lesion, but no grains were detected . The classification of the clinical types of cutaneous nocardiosis is discussed, and the Japanese literature on localized cutaneous nocardiosis is examined. Nippon Jinzo Gakkai Shi, 1992 Mar, 34(3), 325 - 9 {A case of MCTD patient with recurrent hemoperitoneum receiving CAPD who had a successful recovery with an increase in steroids}; Ohtomo Y et al.; A-48-year-old man with end stage renal disease due to MCTD started CAPD in February 1988 . He also had been treated by steroid therapy which was gradually decreased . He had seven hemoperitoneum episodes for ten months and two years since starting CARD . None of episodes were associated with CAPD catheter nor bacillus peritonitis . While he had had long term hemoperitoneum episode in March 1989, he was received no steroid and had leucopenia and low compliments . Therefore he was treated by steroids again . He had no hemoperitoneum episodes for seventeen months since re-steroid therapy . And he recovered from leucopenia and low compliments . It seems that the mechanisms of hemoperitoneum are due to immunological abnormality associated with MCTD. Pharm Res, 1992 Mar, 9(3), 410 - 3 A novel method for determination of sterility of microcapsules and measurement of viability of encapsulated organisms; Kwok KK et al.; A method of determining the viability of microencapsulated microorganisms (Bacillus Calmette Guerin) is reported . This method was also used to measure the effectiveness of aseptic production of microcapsules in maintaining the interior of the microcapsules free from contamination by microorganisms . This method is advantageous over conventional plating methodology, as plating can only determine external contamination of microcapsules and similar devices . It involves the detection of 14CO2, which is generated by the metabolism of 14C-labeled fatty acid in the growth medium by encapsulated microorganisms . The method depends on the semipermeable nature of the microcapsule walls, which allows passage of 14C-palmitic acid and 14CO2 . BCG organisms encapsulated within an alginate-polylysine-alginate microcapsule (5-15 microns) (1) were shown to be viable, and no contaminating organism(s) was present . Methods suitable for the aseptic production and freeze drying of alginate-polylysine-alginate BCG microcapsules, which retain the viability of the organisms, are reported. J Invertebr Pathol, 1992 Mar, 59(2), 186 - 9 Digestion of Bacillus thuringiensis var . israelensis spores by larvae of Aedes aegypti; Khawaled K et al.; The larvicidal activity of Bacillus thuringiensis var . israelensis against mosquitoes and the blackfly is included in parasporal crystalline bodies which are produced during sporulation . Following ingestion, the crystals are solubilized in the larval midgut and induce death within a short time; the spores germinate in the dead larvae and complete a growth cycle . The fate of the spores in surviving live larvae was elucidated by using a nonlarvicidal B . thuringiensis var . israelensis mutant . When introduced as the only food source, spores of this mutant support development to the adult stage of newly hatched Aedes aegypti larvae at a rate directly related to spore concentration . The conclusion that spores of B . thuringiensis var . israelensis are digested in the larval gut was substantiated by following the incorporation of {35S}methionine-labeled spores into larval tissues. J Invertebr Pathol, 1992 Mar, 59(2), 149 - 54 Comparative toxicity of the HD-1 and NRD-12 strains of Bacillus thuringiensis subsp . kurstaki to defoliating forest Lepidoptera; van Frankenhuyzen K et al.; Insecticidal activities of sporulated cultures of the HD-1 and NRD-12 strains of Bacillus thuringiensis subsp . kurstaki were compared against four species of defoliating forest lepidopterans in diet-incorporation assays . There was no difference in LC50 between the two strains to larvae of spruce budworm (Choristoneura fumiferana), gypsy moth (Lymantria dispar), eastern hemlock looper (Lambdina fiscellaria fiscellaria), and whitemarked tussock moth (Orgyia leucostigma), whether expressed as total alkaline soluble protein, activated toxin protein, or International Units as determined by bioassay against Trichoplusia ni . Both strains were consistently more toxic than HD-1-S-1980 when compared on the basis of alkali-soluble protein, but not on the basis of activated toxin or International Units . Hybridization of genomic DNA after restriction with HindIII revealed the presence of all three cryIA toxin genes in each of the isolates used in this study, including HD-1-S-1980, which was previously reported to have lost the cryIA(b) gene. Rev Prat, 1992 Mar 1, 42(5), 606 - 12 {Leprosy . Developmental modalities}; Grossetete G et al.; The very broad clinical spectrum of the Mycobacterium leprae infection is due to the diversity of the underlying immunological and genetic factors . The evolutive modalities of leprosy are mainly determined by a dual pathogenesis: An infectious disease due to a bacillus of low virulence which, even when dead, persists in the body for several years, independently of the antibiotic therapy prescribed . A dysimmune disease maintained by a chronic discharge of antigens . Neuropathies and leprous reactions are still the most troublesome episodes in the course of the disease . They constitute the principal prognostic factor in both pauci- and multibacillary forms of leprosy. Eur J Clin Microbiol Infect Dis, 1992 Mar, 11(3), 240 - 3 A pseudo-outbreak of Methylobacterium mesophilica isolated from patients undergoing bronchoscopy; Flournoy DJ et al.; An unusual, slow growing, pink-pigmented gram-negative bacillus was isolated from bronchoscopy specimens of seven patients over a three-month period . The organism was identified as Methylobacter mesophilica . None of the patients were believed to be infected with Methylobacter mesophilica . The results of environmental cultures showed that the organism was present in tap water from the bronchoscopy room. Scand J Prim Health Care, 1992 Mar, 10(1), 30 - 5 The use of selected interventions in monitoring primary health care implementation in rural Nigeria; Egwu IN; Nigeria's Primary Health Care (PHC)-based health system development aims to strengthen PHC in the local government areas (LGA) through technical planning and implementation that emphasize maternal and child health services . Convenient variables, including expanded programme on immunization (EPI), antenatal care (ANC) utilization and attended births, were selected as interventions to monitor the progress of implementation of PHC activities during 1985-90 in Odukpani LGA . Analysis of available data at the LGA showed that immunization coverage for most EPI antigens increased; ANC services showed increased utilization; health worker-attended births increased as traditional birth deliveries declined during the period . Some of the increases were modest but are considered important . The study offers a pilot approach to monitoring implementation of PHC activities in Odukpani LGA . The implications of the findings for similar studies are discussedPIP: A community health professor analyzed 1986-1989 data on selected primary health care (PHC) services (immunizations, prenatal care, and delivery) to examine the progress of PHC interventions in the rural Odukpani local government area (LGA) in Cross River State, Nigeria . Coverage for the diphtheria-pertussis-tetanus (DPT) and oral poliovirus (OPV) vaccines increased steadily over time (DPT-1 and DPT-2 18-32%; DPT-3 3-28%; OPV-1 18-35%; OPV-2 8-32%; and OPV-3 3-28%) . On the other hand, measles and bacille Calmette-Guerin (BCG) vaccine coverage fell (42-28% and 49-31%, respectively) . Tetanus toxoid coverage was 13% in 1986 and fell to 12% in 1987 and increased above original levels in 1988 (17%) . Odukpani LGA vaccine coverage figures were below national coverage figures . The proportion of all 15-49 year old women using antenatal services fluctuated around 20% during this period, but the percent increase in utilization of antenatal services increased from 0-14% . Overall, the proportion of health worker (HW) attended deliveries increased (e.g., 56-69% during 1988-1989) while the proportion of traditional birth attended (TBA) deliveries decreased (e.g., 45-31% during 1988-1989) . On the other hand, in 1986-1987, TBA deliveries rose (39-47%) while HW attended deliveries fell (62-53%) . Traditional birth attendants did receive training to reduce maternal and child mortality . There was a real possibility of underreporting in this LGA . These results do indicate, however, that the PHC system has effected at least modest increases in specific interventions . This pilot approach can serve as a practical means to monitor implementation of PHC efforts . FEMS Microbiol Lett, 1992 Mar 1, 70(2), 165 - 70 Cloning and nucleotide sequence of the gene encoding the gamma-D-glutamyl-L-diamino acid endopeptidase II of Bacillus sphaericus; Hourdou ML et al.; The gene encoding the Bacillus sphaericus gamma-D-glutamyl-L-diamino acid endopeptidase II, a cytoplasmic enzyme involved in cell sporulation {1}, contains the information for a 271-amino acid protein devoid of a signal peptide . The endopeptidase lacks sequence relatedness with other proteins of known primary structure except that its C-terminal region has significant similarity with the C-terminal region of the 54-kDa P54 protein of Enterococcus faecium, of unknown function {2}. J Am Mosq Control Assoc, 1992 Mar, 8(1), 24 - 31 Impact of the black fly (Diptera: Simuliidae) control agent Bacillus thuringiensis var . israelensis on chironomids (Diptera: Chironomidae) and other nontarget insects: results of ten field trials; Molloy DP; Except for moderate mortality among filter-feeding chironomids, Rheotanytarsus spp., the results of 10 field trials with Bacillus thuringiensis var . israelensis (B.t.i.) indicated a wide margin of safety to the chironomid community and other stream nontarget insects . Mayflies, caddisflies and 2 other types of chironomids, i.e., tube-dwelling (Chironominae) and surface-dwelling, caseless larvae (mix of Chironominae, Diamesinae, Orthocladiinae, Tanypodinae), did not appear to be affected . The susceptibility of filter-feeding chironomids was considerably less than black flies; for example, 4 months of data collected during an operational black fly control program indicated a mean (+/- 95% CI) mortality among Rheotanytarsus larvae of 23 (15-32)% vs . 98 (97-99)% among black flies . Although clearly demonstrating the potential of adverse impact on filter-feeding chironomids in operational black fly programs, these trials also confirmed the narrow impact of B.t.i . on the overall stream insect community. Appl Environ Microbiol, 1992 Mar, 58(3), 815 - 20 Purification and characterization of an endoglucanase from Streptomyces lividans 66 and DNA sequence of the gene; Theberge M et al.; The endoglucanase isolated from culture filtrates of Streptomyces lividans IAF74 was shown to have an Mr of 46,000 and a pI of 3.3 . The specific enzyme activity of 539 IU/mg, determined by the reducing assay method on carboxymethyl cellulose, is among the highest reported in the literature . The cellulase showed typical endo-type activity when reacting on oligocellodextrins . Optimal enzyme activity was obtained at 50 degrees C and pH 5.5 . The kinetic constants for this endoglucanase, determined with carboxymethyl cellulose as the substrate, were a Vmax of 24.9 IU/mg of enzyme and a Km of 4.2 mg/ml . Activity was found against neither methylumbelliferyl- nor p-nitrophenyl-cellobiopyranoside nor with xylan . The DNA sequence contains one possible reading frame validated by the N terminus of the mature purified protein . However, neither ATG nor GTG starting codons were identified near the ribosome-binding site . A putative TTG codon was found as a good candidate for the start codon . Comparison of the primary amino acid sequence of the endoglucanase of S . lividans revealed that the N terminus contains a bacterial cellulose-binding domain . The catalytic domain at the C terminus showed similarity to endoglucanases from a Bacillus sp . Thus, the endoglucanase CelA belongs to family A of cellulases as described before (N . R . Gilkes, B . Henrissat, D . G . Kilburn, R . C . Miller, Jr., and R . A . J . Warren, Microbiol . Rev . 55:303-315, 1991. J Appl Bacteriol, 1992 Mar, 72(3), 188 - 94 Antimicrobial properties of secretions from the metapleural glands of Myrmecia gulosa (the Australian bull ant); Veal DA et al.; Myrmecia gulosa (Australian bull ant) produce secretions from their metapleural exocrine glands which have broad spectrum antimicrobial properties . Such secretions are probably of importance in disease control in bull ant communities . These antimicrobial secretions are stable at 100 degrees C, resistant to proteolytic enzymes and are active over a wide pH range . Of the organisms tested only endospores of Bacillus cereus were found to be resistant . The antimicrobial agent(s) are absorbed by cells and result in cell lysis . The secretions do not interfere with any growth-related processes . These observations demonstrate that insects may be a source of novel antimicrobial agent(s). Nippon Hinyokika Gakkai Zasshi, 1992 Mar, 83(3), 368 - 73 {Histological study on intravesical instillation of Bacillus Calmette Guerin (BCG) and adriamycin for BBN-induced bladder tumor in rats}; Tanaka S et al.; We evaluated histological changes induced by intravesical instillation of Bacillus Calmette Guerin (BCG) and doxorubucin hydorochloride (Adriamycin, ADM) for transitional cell carcinoma of the experimental bladder tumor . We administered rats N-butyl-N (4-hydroxybutyl) nitrosamine (BBN) by mixing it in the drinking water, and performed intravesical instillation of BCG, ADM or physiological saline solution as control in the 16th and 17th weeks and sacrificed in the 18th week after BBN exposure, and observed histological changes by light microscopy and electron microscopy . In the BCG group, the enlargement of the intercellular spaces of superficial tumor cells was found, and electron microscopic findings revealed a decrease or disappearance of the junctional complex . In the ADM group, hydrolytic and vacuolated cytoplasm was revealed . But such change as that in the BCG group was not found in the ADM and control groups . These results indicate that BCG may enlarge the intercellular spaces by changing the junctional complex, and lead to desquamate the tumor cells from the superficial cell layer. Infect Control Hosp Epidemiol, 1992 Mar, 13(3), 160 - 4 Clustered bacteremias in a hemodialysis unit: cross-contamination of blood tubing from ultrafiltrate waste; Longfield RN et al.; OBJECTIVE: To determine the cause of clustered bacteremias occurring among chronic hemodialysis patients . DESIGN: A retrospective investigation of clinical and laboratory records with direct observation of dialysis facilities and technique . Bacterial blood isolates were identified and compared with environmental isolates . SETTING: The 11-station chronic hemodialysis unit that serves approximately 50 patients in a 450-bed military hospital . PATIENTS: Hemodialysis unit patients with aerobic gram-negative bacillus or Enterococcus casseliflavus blood isolates between April 1988 and February 1990 . RESULTS: The recovery and species identification of the unique isolate, E casseliflavus, from 2 index cases of bacteremia in February 1990 helped identify the cluster and demonstrated its protracted course . Dialysis blood tubing was contaminated with ultrafiltrate waste during dialyzer setup . INTERVENTION: Bacteremias were controlled by halting the practice of attaching the venous tubing directly to a waste container while priming the membrane, by emphasizing glove changes and handwashing after contact with ultrafiltrate waste and by daily decontamination of ultrafiltrate waste bags . CONCLUSIONS: We recommend that other hemodialysis units institute these interventions. Mol Microbiol, 1992 Mar, 6(5), 563 - 7 DNA in dormant spores of Bacillus species is in an A-like conformation; Setlow P; The DNA in dormant spores of Bacillus species is associated with alpha/beta-type small, acid-soluble proteins (SASP), which are double-stranded DNA-binding proteins whose amino acid sequence has been highly conserved in evolution . In vitro these proteins bind most strongly to DNA which readily adopts an A-like conformation, as binding of alpha/beta-type SASP causes DNA to assume an A-like conformation . As predicted by this conformational change in DNA, binding of alpha/beta-type SASP to relaxed but covalently closed plasmid DNA results in the introduction of a large number of negative supercoils . Associated with the conformational change in DNA brought about by alpha/beta-type SASP binding is a change in its photochemistry such that ultraviolet irradiation does not generate pyrimidine dimers, but rather a thyminyl-thymine adduct termed spore photoproduct (SP) . The latter two properties of DNA complexed with alpha/beta-type SASP in vitro are similar to those of DNA in dormant spores of Bacillus species in which: (i) plasmid DNA has a much higher number of negative supercoils than plasmid in growing cells; and (ii) ultraviolet irradiation produces SP and no pyrimidine dimers, while only pyrimidine dimers are formed in growing cells . During sporulation these changes in the properties of spore DNA take place in parallel with synthesis of alpha/beta-type SASP, and the magnitude of the changes is greatly reduced in mutants that make low amounts of these proteins . A straightforward interpretation of these data is that DNA in dormant spores of Bacillus species is in an A-like conformation.(ABSTRACT TRUNCATED AT 250 WORDS) Int J Biochem, 1992 Mar, 24(3), 471 - 6 Purification and partial characterization of the gamma-D-glutamyl-L-di-amino acid endopeptidase II from Bacillus sphaericus; Bourgogne T et al.; 1 . A gamma-D-glutamyl-L-di-amino acid endopeptidase II (EC3.4.-.-) active on the peptide moieties of some bacterial peptidoglycans has been purified to homogeneity from the sporulation medium and from the spores of Bacillus sphaericus . 2 . Enzyme from both sources showed a single protein band (Mr 28,000) by polyacrylamide gel electrophoresis under denaturing conditions . It is an acidic protein (pI 4.1) . Kinetic studies have shown a Km value of 0.24 mM and an apparent Vmax of 8.3 mumol min-1 mg-1 with the pentapeptide L-Ala-gamma-D-Glu-L-Lys-D-{14C}Ala-D-{14C}Ala as substrate . 3 . The enzyme was inhibited by p-hydroxymercuribenzoate, a sulfhydryl inhibitor . 4 . The 38-residue N-terminal region was sequenced . It may be useful to construct a nucleotide probe for the research of the gene encoding this enzyme. Arch Intern Med, 1992 Mar, 152(3), 602 - 6 Rochalimaea henselae causes bacillary angiomatosis and peliosis hepatis; Slater LN et al.; BACKGROUND--Recent studies have demonstrated that a newly described agent of persistent bacteremia, Rochalimaea henselae, and the agent of bacillary angiomatosis are both closely related to Rochalimaea quintana . Bacillary peliosis hepatis seemed likely to have the same etiologic agent as bacillary angiomatosis . We sought these pathologic changes in patients from whom R henselae was cultivated . METHODS--For two patients whose histopathologic findings we reviewed, additional light and electron microscopy were performed . Their bacterial isolates were compared by electrophoretic patterns of outer membrane proteins, restriction endonuclease digestion patterns of DNA, and reaction with murine antiserum . RESULTS--A previously reported human immunodeficiency virus-infected man with persistent bacteremia due to R henselae was found to have bacillary peliosis hepatis . Rochalimaea henselae was also isolated from the spleen of a woman receiving immunosuppressive therapy after allogeneic renal transplantation . She had developed fever, liver and spleen nodules, and periaortic lymphadenopathy . Bacillary peliosis of her liver and spleen, as well as bacillary angiomatosis of liver, spleen, and a lymph node, were found . The bacterial isolates had comparable electrophoretic patterns of outer membrane proteins and of restriction endonuclease-digested DNA, which differed from the respective patterns of R quintana . Murine antisera raised to the first isolate reacted strongly with the second by means of immunoblot and immunofluorescence techniques, while reacting only weakly with R quintana . CONCLUSION--Rochalimaea henselae, recently recognized to cause persistent fever and bacteremia in immunocompetent and immunocompromised persons, also causes bacillary angiomatosis and parenchymal bacillary peliosis. Am Rev Respir Dis, 1992 Mar, 145(3), 621 - 5 Effect of bacille Calmette-Guérin vaccination on tuberculin reactivity; Menzies R et al.; The effect on tuberculin reactivity of bacille Calmette-Guerin vaccination (BCG-V) given 10 to 25 yr earlier was measured among schoolchildren and young adults in Montreal . Of a total eligible population of 8,264 persons, 5,952 were tuberculin tested (72%) . The BCG-V status could be verified for 4,629 of 4,969 (93%) participants born in Quebec . Of these, 1,511 (33%) had received BCG-V, 66% of whom had been vaccinated once only in infancy, 23% of whom had been vaccinated once but after infancy, and 11% of whom had been vaccinated twice . Among those vaccinated in infancy, 7.9% had significant tuberculin reactions, compared with 18% among those vaccinated between 1 and 5 yr of age, and 25.4% among those vaccinated after the age of 5 (p less than 0.001) . Among those receiving BCG-V in infancy, the prevalence of tuberculin reactions of 10 mm was similar to the nonvaccinated subjects, after accounting for differences in age and socioeconomic status . Among those vaccinated after infancy, tuberculin reactivity was not significantly affected by interval from BCG-V until tuberculin testing nor by repeat BCG-V . We conclude that after an interval of 10 to 25 yr the most important determinant of effect of BCG-V on tuberculin reactivity is age when vaccinated . A significant tuberculin reaction among those who have received BCG-V should be interpreted in light of the age when vaccinated and the expected prevalence of tuberculous infection in the population tested. Infect Immun, 1992 Mar, 60(3), 937 - 43 Deformation factor: an extracellular protein synthesized by Bartonella bacilliformis that deforms erythrocyte membranes; Mernaugh G et al.; Bartonella bacilliformis, a hemotropic bacterium and the causative agent of the human disease bartonellosis, when incubated in a tryptone-based medium produces an extracellular factor, termed deformation factor (DF), which induces extensive indentations and trenches in trypsinized erythrocyte membranes . The factor is stable during storage at 4 degrees C . It can be inactivated by proteases or brief heating to 70 to 80 degrees C, can be precipitated by ammonium sulfate, is nondialyzable, and is retained by membranes with a 30,000-molecular-weight cutoff . These properties suggest that DF is probably a protein . Incubation of erythrocytes with phospholipase D renders them resistant to deformation by DF. J Urol, 1992 Mar, 147(3), 738 - 42 Antibiotic and steroid therapy of massive systemic bacillus Calmette-Guerin toxicity; DeHaven JI et al.; Intravesical bacillus Calmette-Guerin (BCG) is the most effective treatment of carcinoma in situ available today and is superior to chemotherapy in the prevention of bladder tumor recurrence . While therapy is generally well tolerated, serious and even life threatening toxicity can occur . Treatment options for serious infection include isoniazid, rifampin, ethambutol, and cycloserine, but shock may also be secondary to hypersensitivity and require the addition of corticosteroids . The morbidity and mortality of systemically BCG-infected mice treated with single and combined antimicrobial and/or corticosteroid therapies was evaluated . BCG immunized mice were unable to survive doses of BCG which were uniformly tolerated in naive mice . The addition of cycloserine increased survival in mice treated with isoniazid and rifampin, but optimal survival was achieved with isoniazid, rifampin, and prednisolone . These experimental results support the previously reported clinical success of isoniazid, rifampin and prednisolone in patients with septic BCG reactions. J Urol, 1992 Mar, 147(3), 695 - 7 Hypersensitivity systemic reaction following intravesical bacillus Calmette-Guerin: successful treatment with steroids; Molina JM et al.; Intravesical bacillus Calmette-Guerin (BCG) is an effective treatment for superficial bladder carcinoma . Serious complications, including disseminated BCG infection, are infrequent . We report a case of granulomatous hepatitis with pneumonitis following intravesical administration of BCG . Cultures for mycobacteria were negative in sputum, bronchoalveolar lavage, liver and blood specimens . All symptoms disappeared within days after steroid therapy . Hypersensitivity reaction should be considered in patients with systemic symptoms after immunotherapy with BCG. J Urol, 1992 Mar, 147(3), 601 - 5 Management of local bacillus Calmette-Guerin failures in superficial bladder cancer; Klein EA et al.; We attempt to define the treated natural history of patients with superficial bladder tumors (stages Ta, TIS and T1) managed with intravesical bacillus Calmette-Guerin (BCG) to determine the best form of treatment for locally recurrent tumors . The management and survival of 41 patients who failed BCG within the bladder or prostatic urethra and who subsequently were treated with a variety of secondary therapies are reviewed . Our aim was to assess the role of several independent clinical variables on the rate of death from bladder cancer . Of the 41 patients 6 (15%) died . Univariate statistical analysis identified early involvement of the prostatic urethra and the presence of superficially invasive (stage T1) tumor at initial treatment with BCG as factors having an adverse effect on survival . A multivariate statistical model revealed that patients with early prostatic urethral involvement and the presence of superficially invasive (stage T1) tumor at diagnosis had the highest risk of death from bladder cancer . The reason for change in therapy at failure of BCG, stage of the tumor at BCG failure, occurrence of upper tract tumors and early versus delayed radical cystectomy had no impact on survival . The results suggest that not all tumors that recur after BCG are destined to proceed to muscle invasion or metastases, and that some patients may be managed safely by repeated endoscopic resection and intravesical therapy with cystectomy delayed until objective progression is evident . Such an approach can yield survival equal to that in patients treated with early cystectomy and may result in longer intervals of bladder preservation in a select subset of patients who fail BCG locally. J Urol, 1992 Mar, 147(3), 596 - 600 Incidence and treatment of complications of bacillus Calmette-Guerin intravesical therapy in superficial bladder cancer; Lamm DL et al.; Intravesical therapy with bacillus Calmette-Guerin (BCG) has proved to be more effective in the prophylaxis and treatment of superficial bladder tumors and carcinoma in situ than most chemotherapeutic agents . Compared to intravesical chemotherapy, instillations with BCG provoke more local and systemic reactions . In addition to the commonly induced granulomatous inflammatory changes in the bladder, which produce irritative symptoms, this therapy may cause systemic side effects varying from mild malaise and fever to, in rare instances, life-threatening or fatal sepsis . We report the incidence and varieties of toxicities in 2,602 patients treated with intravesical BCG . Side effects are classified according to local and systemic toxicity . Treatment options vary according to the severity of toxicity from delaying or withholding instillations to treatment with antituberculous drugs for up to 6 months . In general, 95% of the patients have no serious side effects . Recognition of risk factors, particularly traumatic catheterization or concurrent cystitis, that result in systemic BCG absorption, as well as the prompt and appropriate treatment of early side effects should significantly decrease the incidence of severe toxicity. Gastroenterology, 1992 Mar, 102(3), 1065 - 70 Bacillary peliosis hepatis as a cause of acute anemia in a patient with the acquired immunodeficiency syndrome; Garcia-Tsao G et al.; A 33-year-old white homosexual man, infected with the human immunodeficiency virus, presented with acute anemia and thrombocytopenia not responsive to transfusions or to treatment with steroids and intravenous gamma-globulin . Hematologic workup was compatible with peripheral sequestration or loss of blood cells; however, there was no evidence of gastrointestinal or other sources of hemorrhage, and the only significant finding was a progressive liver enlargement . An abdominal computerized tomographic scan showed a massive homogeneous liver without focal lesions, a very small amount of ascites, and no retroperitoneal fluid collections . A transjugular liver biopsy specimen showed the cystic, blood-filled cavities characteristic of peliosis hepatis . Cavities varied in size, all contained pooled erythrocytes, and some had areas suggestive of thrombi in various stages of organization . Bacteria similar in morphology to those described in bacillary peliosis hepatis were seen in the peliotic spaces . The clinical picture began resolving shortly after treatment with zidovudine and ampicillin/sulbactam was started and had totally resolved 6 months after presentation . This case shows that bacillary peliosis hepatis is a reversible entity that may produce acute sequestration of blood in the liver. Prikl Biokhim Mikrobiol, 1992 Mar-Apr, 28(2), 173 - 7 {BME 361 I--a new site-specific deoxyribonuclease type II from Bacillus megaterium 361}; Lebedev LR et al.; Bme 361 I, a new site-specific type II deoxyribonuclease, was purified from Bacillus megaterium 361 by chromatography on phosphocellulose P 11 and hydroxylapatite . The enzyme recognizes and cleaves the nucleotide sequence 5'-GG decreases CC-3' in double-strand DNA . Thus it is a true isoschizomer of deoxyribonucleases Hae III and BspR I. Bioconjug Chem, 1992 Mar-Apr, 3(2), 176 - 81 Monoclonal antibody-beta-lactamase conjugates for the activation of a cephalosporin mustard prodrug; Svensson HP et al.; Cephalosporin mustard (CM) was designed as an anticancer prodrug that could be activated in a site-specific manner by monoclonal antibody-beta-lactamase conjugates targeted to antigens present on tumor cell surfaces . Purified beta-lactamases from Bacillus cereus (BC beta L) and Escherichia coli (EC beta L) catalyzed the release of phenylenediamine mustard (PDM) from CM through a fragmentation reaction which occurs after the beta-lactam ring of CM is hydrolyzed . The Km and Vmax values were 5.7 microM and 201 mumol/min per mg for BC beta L and 43 microM and 29 mumol/min per mg for EC beta L, respectively . Conjugates of BC beta L were prepared by combining the F(ab')2 fragments of the maleimide-substituted monoclonal antibodies L6 and 1F5 with thiolated BC beta L . The conjugates showed little loss in enzymatic activity and bound nearly as well as the unmodified F(ab')2 monoclonal antibodies to antigens expressed on the H2981 human lung adenocarcinoma cell line (L6 positive, 1F5 negative) . PDM was approximately 50-fold more cytotoxic than CM to H2981 cells . Treatment of the cells with L6-BC beta L followed by CM resulted in a level of cytotoxic activity that was comparable to that of PDM . This was most likely due to activation of CM by conjugate that bound to cell-surface antigens, since pretreatment of H2981 cells with BC beta L or 1F5-BC beta L enhanced the activity of CM to a lesser extent . Thus, we have shown that CM is a prodrug, and that it can be activated with immunological specificity by a monoclonal antibody-beta-lactamase conjugate. J Med Entomol, 1992 Mar, 29(2), 232 - 5 Relationship between selected bacteria and the growth of immature house flies, Musca domestica, in an axenic test system; Schmidtmann ET et al.; To investigate the relationship between immature (maggot) house flies, Musca domestica, and bacteria, we compared the development of sterile first-instar maggots in each of 10 pure blood agar cultures of bacteria with growth on sterile blood agar (negative control) and on standard house fly rearing medium (positive control) . Nine species of bacteria representing gram-negative and gram-positive rods, coccoid, and micrococcoid cell types supported house fly growth on blood agar . One bacterium, a strain of Bacillus cereus, inhibited maggot growth . The percent pupation for maggots that developed in the presence of eight of nine bacteria (range, 41-69%), was significantly greater than in sterile blood agar (0-5%), and did not differ significantly from maggot growth in the rearing medium (50-90%) . Average pupal weight for maggots that developed on blood agar with bacteria ranged from 19 to 21 mg, a reflection of favorable growth conditions . Average pupal weight in the presence of three bacteria (19.9, 19.4, and 19.4 mg) was significantly less than respective pupal weights in house fly rearing medium (24.0, 22.3, and 22.1 mg), but there was no difference in average pupal weight with six bacteria and the house fly rearing medium . These findings illustrate that bacteria or their metabolic products are essential as nutrients for house fly maggot growth in blood agar; a wide variety of adventitious bacteria can contribute to the suitability of an organic substrate for maggot growth; and a naturally occurring isolate of B . cereus limits house fly maggot growth in blood agar, a relationship that has not been reported previously. Immunology, 1992 Mar, 75(3), 414 - 9 CD4+ T cells are required for antigen-specific recruitment of neutrophils by BCG-immune spleen cells; Appelberg R; Mycobacterium bovis bacillus Calmette-Guerin (BCG)-immune spleen cells co-inoculated into the peritoneal cavity of normal mice with BCG sonicate protein as antigen could induce an antigen-specific recruitment of neutrophils, dependent on the antigen dose and cell number . This response was significantly reduced by anti-T lymphocyte and anti-CD4 treatment of the immune spleen cells prior to the inoculation . Removal of adherent or phagocytic cells or lysis of B cells, had no significant effect . Killing of dividing cells in the splenic population induced a slight reduction in the ability of spleen cells to recruit neutrophils . M . avium sonicate protein was also able to induce BCG-immune spleen cells to mobilize neutrophils but bovine serum albumin, Listeria monocytogenes cytosolic protein and 65,000 MW heat shock protein were not . These results show that CD4+ T cells are able to induce neutrophil recruitment in an antigen-specific way during a mycobacterial infection. Arthritis Rheum, 1992 Mar, 35(3), 270 - 81 Mycobacteria and human heat shock protein-specific cytotoxic T lymphocytes in rheumatoid synovial inflammation; Li SG et al.; OBJECTIVE . To study the cytotoxic capacity of mycobacteria-specific T lymphocyte lines and clones from sites of inflammation in patients with rheumatoid arthritis (RA) . We also studied antigen specificity, surface phenotype, expression of T cell receptors (TCR), and HLA restriction . METHODS . Autologous macrophages (M phi) from the synovial membrane (SM), synovial fluid (SF), or peripheral blood (PB) were used as target cells in cytotoxicity assays . RESULTS . All SM and SF cell lines tested thus far have shown specific lysis of the autologous M phi from SM or PB that had been pulsed with BCG (bacillus Calmette-Guerin), but no cytotoxicity when the targets were pulsed with irrelevant antigens such as tetanus toxoid and Chlamydia . Both CD4+ and CD8+ cells were shown to be involved in the specific cytolysis . The majority of the cytotoxic T lymphocyte (CTL) lines were TCR alpha/beta + cells . However, both TCR alpha/beta + and TCR gamma/delta + clones (TCR delta 1+) from one RA patient showed antigen-specific lysis . Antigen-specific recognition by a number of CTL lines and clones generated from SF and SM was restricted by HLA-DR molecules . Two Mycobacterium bovis 65-kd heat shock protein (HSP)-specific TCR alpha/beta + SF T cell clones also lysed M phi that had been pulsed with a recombinant human 65-kd HSP . CONCLUSION . Joint inflammation and destruction might be partly attributable to a cross-reaction of mycobacteria-induced cytotoxic T cells with self HSP. J Biotechnol, 1992 Mar, 23(1), 19 - 34 Construction of an Escherichia coli export-affinity vector for expression and purification of foreign proteins by fusion to cyclomaltodextrin glucanotransferase; Hellman J et al.; A novel export-affinity fusion vector employing the gene encoding cyclomaltodextrin glucanotransferase (CGTase; cgt) from Bacillus circulans var . alkalophilus (ATCC 21783) is described . CGTase binds to various sugar polymers, which makes it simple to purify it to near homogeneity in a single step . The CGTase fusion protein vector was constructed by deleting the translational stop codons from the gene encoding CGTase (cgt) by in vitro mutagenesis . As models, genes encoding Escherichia coli alkaline phosphatase (APase; phoA) and Bacillus stearothermophilus (ATCC 12980) alpha-amylase (BStA; amy) were fused to cgt . Overexpression of wild type CGTase and the hybrid proteins under the control of the lac promoter caused a 'leaky phenotype' in E . coli, the outer membrane became permeable, which enabled the adsorption of the fusion proteins directly from the culture medium onto alpha-cyclodextrin (alpha-CD) coupled agarose . The hybrid proteins were eluted from the column with alpha-CD solution under mild conditions at pH 7.5 . The CGTase-APase' fusion had a good in vivo stability, whereas the CGTase-BStA' was less stable . In the latter case, according to protein sequencing, the proteolytically sensitive site was on the BStA' side of the fusion . The C-terminus of CGTase was stable against proteolysis as shown by narrow pH range isoelectric focusing . The fused enzymes retained their biological activities. Biosci Biotechnol Biochem, 1992 Mar, 56(3), 448 - 53 Action pattern of Bacillus sp . no . 7-M chitosanase on partially N-acetylated chitosan; Izume M et al.; The hydrolyzate of partially N-acetylated chitosan by Bacillus sp . No . 7-M chitosanase was separated by gel filtration on Bio-Gel P-2 . Sugar compositions and sequences of the oligosaccharides were identified by exo-splitting with beta-GlcNase, fast atom bombardment mass spectroscopy, and proton NMR spectroscopy . In addition to chitooligosaccharides, (GlcN)2, (GlcN)3, and (GlcN)4, hetero-chitooligosaccharides such as (GlcN)2.GlcNAc.(GlcN)2, GlcN.GlcNAc.(GlcN)3, (GlcN)2.GlcNAc.(GlcN)3, and GlcN.GlcNAc.(GlcN)4 were detected . These results indicate that Bacillus sp . No . 7-M chitosanase is absolutely specific toward the GlcN.GlcN bonds in partially N-acetylated chitosan and at least three GlcN residues were necessary to the hydrolysis of chitosan by chitosanase. Biotechnology (N Y), 1992 Mar, 10(3), 292 - 6 Production of active Bacillus licheniformis alpha-amylase in tobacco and its application in starch liquefaction; Pen J et al.; As a first example of the feasibility of producing industrial bulk enzymes in plants, we have expressed Bacillus licheniformis alpha-amylase in transgenic tobacco, and applied the seeds directly in starch liquification . The enzyme was properly secreted into the intercellular space, and maximum expression levels of about 0.3% of total soluble protein were obtained . No apparent effect of the presence of the enzyme on plant phenotype was observed . The molecular weight of the enzyme produced in tobacco was around 64 kD . The difference, compared to 55.2 kD for the bacterial enzyme, was found to result from complex-type carbohydrate chains attached to the protein . Application studies on the liquefaction of starch were done with transgenic seeds containing the recombinant alpha-amylase . The resulting hydrolysis products were virtually identical with those obtained from degradation with alpha-amylase from Bacillus licheniformis. J Leukoc Biol, 1992 Mar, 51(3), 225 - 9 Effects of in vivo T lymphocyte subset depletion on mycobacterial infections in mice; Flory CM et al.; The relative importance of CD4+ and CD8+ T cell subsets in the expression of acquired resistance to systemic infection by Mycobacterium kansasii was determined . T cell subsets were depleted in thymectomized C57BL/6 mice by the intravenous administration of monoclonal antibodies directed against the relevant T cell determinants . Depletion of the CD4+ subset exacerbated the severity of the infection in intravenously challenged mice . This effect was apparent in the first 2 weeks of the infection and persisted throughout the 12 weeks of the study . On the other hand, depletion of the CD8+ cells had no apparent effect on the growth curves . Infections by Mycobacterium tuberculosis Erdman or bacille Calmette-Guerin (BCG) Pasteur were also substantially enhanced by CD4 depletion, but not by the depletion of CD8+ cells . The effect of subset depletion on infections by M . tuberculosis and BCG was examined in both innately susceptible C57BL/6 mice and innately resistant B6D2 mice. Mol Gen Genet, 1992 Mar, 232(1), 162 - 5 Sensitivity to plating of Escherichia coli cells expressing the cryA gene from Bacillus thuringiensis var . israelensis; Douek J et al.; The gene (cytA) coding for the 27 kDa polypeptide of the Bacillus thuringiensis var . israelensis mosquito larvicidal delta-endotoxin, was cloned into a plasmid containing the T7 bacteriophage promoter . The plasmid was used to transform an Escherichia coli strain containing the T7 RNA polymerase gene 1, under the control of lacP . Loss of colony-forming ability without substantial lysis, associated with immediate inhibition of DNA synthesis, was observed after induction of transformed cells . The cytA gene product may kill E . coli cells by disrupting their chromosome replicating apparatus. J Biol Chem, 1992 Feb 15, 267(5), 3115 - 21 Location of a Bombyx mori receptor binding region on a Bacillus thuringiensis delta-endotoxin; Lee MK et al.; Receptor binding studies were performed with 125I-labeled trypsin-activated insecticidal toxins, CryIA(a) and CryIA(c), from Bacillus thuringiensis on brush-border membrane vesicles (BBMV) prepared from Bombyx mori larval midgut . Bioassays were performed by gently force feeding B . mori with diluted toxins . CryIA(a) toxin (LD50; 0.002 micrograms) was 200 times more active against B . mori larvae than CryIA(c) toxin (LD50; 0.421 micrograms) and showed high-affinity saturable binding . The Kd and the binding site concentration for CryIA(a) toxin were 3.5 nM and 7.95 pmol/mg, respectively . CryIA(c) toxin (Kd, 50.35 nM; Bmax, 2.85 pmol/mg) did not demonstrate high-affinity binding to B . mori BBMV . Control experiments with CryIA(a) and CryIA(c) toxins revealed no binding to mouse small intestine BBMV and nonspecific binding to pig kidney BBMV . These data provide evidence that binding to a specific receptor on the membrane of midgut epithelial cells is an important determinant with respect to differences in insecticidal spectrum of insecticidal crystal proteins . To locate a B . mori receptor binding region on the CryIA(a) toxin, homologous and heterologous competition binding studies were performed with a set of mutant proteins which had previously been used to define the B . mori "specificity domain" on this toxin (Ge, A . Z., Shivarova, N . I., and Dean, D . H . (1989) Proc . Natl . Acad . Sci . U.S.A . 86, 4037-4041) . These mutant proteins have had regions of their genes reciprocally exchanged with the cryIA(c) gene . A B . mori receptor binding region on CryIA(a) toxin includes the amino-terminal portion of the hypervariable region, amino acids 332-450, which is identical to the previously described B . mori specificity determining region . These data provide direct evidence that delta-endotoxins contain a tract of amino acids that comprise a binding region and as a results determines the specificity of a toxin. Biochem Biophys Res Commun, 1992 Feb 14, 182(3), 1309 - 12 THR246 mutations decrease substrate inhibition in lactate dehydrogenase; Sakowicz R et al.; Threonine 246 in Bacillus stearothermophilus L-lactate dehydrogenase has been changed to valine, serine, and alanine by site-directed mutagenesis . Kinetic analyses show a decrease in substrate inhibition for pyruvate reduction with the T246S mutant and virtual elimination of substrate inhibition for the T246A and T246V mutants . The results indicate that the absence of substrate inhibition in the 246A/V-catalyzed reactions is due to the elimination of a key hydrogen bond between the hydroxyl group of threonine and pyruvate in the wild-type complex that is an important contributor in the formation of the abortive enzyme-NAD(+)-pyruvate complex responsible for substrate inhibition. Cell Immunol, 1992 Feb, 139(2), 493 - 504 Impairment of macrophage activation and granuloma formation by protein deprivation in mice; Reynolds JV et al.; Protein-calorie malnutrition predisposes to infection by intracellular pathogens, but the basis for this predisposition is unclear . We studied the effect of protein deprivation on mouse peritoneal macrophage function and on granuloma formation during infection by bacille Calmette-Guerin (BCG) . Injection of lipopolysaccharide (LPS) to induce inflammation elicited fewer peritoneal cells from mice fed a 2.5% protein diet than from mice fed an isocaloric chow in which protein calories constituted 24% of the total . LPS-elicited macrophages from protein-deprived mice demonstrated a reduction in spreading, total cell protein, cell lactate dehydrogenase, and release of superoxide anion (O2-) in response to stimulation . Priming in vitro by interferon (IFN)-gamma for enhanced release of O2- was also significantly impaired in protein-deprived mice . This defect was reversible by repletion with 24% protein diet for 10 days . Impairment of macrophage function in protein-deprived mice was further evidenced by an impaired capacity to express Ia antigen in response to IFN-gamma and by reduced production of IL-1 activity in response to LPS . Infection by BCG in protein-deprived mice was characterized by impaired granuloma development in liver, lungs, and spleen . Thus, in this model, protein deprivation significantly impaired macrophage activation, as assessed by morphologic, metabolic, and functional criteria . This impairment might compromise immune effector mechanisms dependent on macrophage activation, including rejection of intracellular pathogens. FEBS Lett, 1992 Feb 10, 297(3), 226 - 8 Proteolysis of Bacillus stearothermophilus IF2 and specific protection by fMet-tRNA; Severini M et al.; Translation initiation factor IF2 from Bacillus stearothermophilus (741 amino acids, Mr 82,043) was subjected to trypsinolysis alone or in the presence of fMet-tRNA . The initiator tRNA was found to protect very efficiently the Arg308-Ala309 bond within the GTP binding site of IF2 and, more weakly, three bonds (Lys146-Gln147, Lys154-Glu155 and Arg519-Ser520) . The first two are located at the border between the non-conserved, dispensable (for translation) N-terminal portion and the conserved G-domain of the protein, the third is located at the border between the G- and C-domains . Since IF2 is known to interact with fMet-tRNA through its protease-resistant C- (carboxyl terminus) domain, the observed protection suggests that, upon binding of fMet-tRNA, long-distance tertiary interactions between the IF2 domains may take place. J Mol Biol, 1992 Feb 5, 223(3), 801 - 10 Role of residue Glu152 in the discrimination between transfer RNAs by tyrosyl-tRNA synthetase from Bacillus stearothermophilus; Vidal-Cros A et al.; Residue Glu152 of tyrosyl-tRNA synthetase (TyrTS) from Bacillus stearothermophilus is close to phosphate groups 73 and 74 of tRNATyr in the structural model of their complex . TyrTS(E152A), a mutant synthetase carrying the change of Glu152 to Ala, was toxic when overproduced in Escherichia coli . The toxicity strongly increased with the growth temperature . It was measured by the ratios of the efficiencies with which the producing cells plated in induced or repressed conditions and at 30 degrees C or 37 degrees C . TyrTS(E152Q), TyrTS(E152D) and the wild-type synthetase were not toxic in conditions where TyrTS(E152A) was toxic . The toxicity of TyrTS(E152A) was abolished by additional mutations of the synthetase that prevent the binding of tRNATyr but not by a mutation that prevents the formation of Tyr-AMP . Because TyrTS(E152A) was active for the aminoacylation of tRNATyr, its toxicity could only be due to faulty interactions with non-cognate tRNAs, either their non-productive binding or their mischarging with tyrosine . TyrTS(E152A) and TyrTS(E152Q) mischarged tRNAPhe and tRNAVal in vitro with tyrosine unlike TyrTS(E152D) or the wild-type enzyme . Thus, several features of the side-chain in position 152 of TyrTS, including its negative charge, are important for the rejection of non-cognate tRNAs . TyrTS(E152A), TyrTS(E152D) and TyrTS(E152Q) had similar steady-state kinetics parameters for the charging of tRNATyr with tyrosine in vitro, with kcat/KM ratios improved 2.5 times relative to the wild-type synthetase . We conclude that the side-chain of residue Glu152 weakens the binding of TyrTS to tRNATyr and prevents its interaction with non-cognate tRNAs. J Biol Chem, 1992 Feb 5, 267(4), 2311 - 7 Localized mutagenesis defines regions of the Bacillus thuringiensis delta-endotoxin involved in toxicity and specificity; Wu D et al.; Bacillus thuringiensis produces a variety of delta-endotoxins which bind to specific receptors in insect larval midguts . Following insertion into the membrane there is an alteration of ion flux culminating in osmotic lysis . Mutagenic oligonucleotides were used to define regions in one of these toxins involved in specificity and toxicity . One region is highly conserved among all toxins sequenced to date and many mutations resulted in loss of toxicity for three test Lepidoptera . The mutant toxins had lost the capacity to inhibit K(+)-dependent amino acid transport into larval midgut vesicles, but there was no effect on their ability to compete with wild type toxin for binding . The results are consistent with this amphiphilic helical region of the toxin being essential for toxicity . A second mutagenized region overlapped a portion of another potential amphiphilic helix . Mutations of only 2 residues, Ala-92 and Arg-93, resulted in loss of toxicity for two lepidopteran larvae but some activity remained for a third . The A92D mutant toxin competed with the wild type toxin for binding to vesicles prepared from midguts from the sensitive but not from the insensitive larvae . Decreased toxicity was also found when this mutation was transferred to two other related protoxin genes . A number of mutations of each of these residues was analyzed and selective loss of toxicity correlated with the absence of a positive charge . Despite being distal from the presumptive specificity domain, 1 or both of these residues must have an important role in the specific binding of toxins. FEBS Lett, 1992 Feb 3, 297(1-2), 164 - 6 Introduction of a free cysteinyl residue at position 68 in the subtilisin Savinase, based on homology with proteinase K; Bech LM et al.; Two subfamilies of the subtilisins, distinguished by the presence or absence of a free cysteinyl residue near the essential histidyl residue of the catalytic triad, are known . In order to evaluate the significance of the presence of this -SH group a cysteinyl residue has been introduced by site-directed mutagenesis into the cysteine-free subtilisin-like enzyme from Bacillus lentus, i.e . Savinase . The free cysteine affects the enzyme activity only slightly but renders it sensitive to mercurials presumably due to an indirect effect . The results indicate that the -SH group is not involved in catalysis. J Urol, 1992 Feb, 147(2), 482 - 5 Enhancement of bacillus Calmette-Guerin attachment to the urothelium by removal of the rabbit bladder mucin layer; Badalament RA et al.; It has been established that the urothelial mucin layer functions as a bacterial anti-adherence factor . Intravesical Bacillus Calmette-Guerin is used to treat patients with superficial bladder cancer . The proposed mechanism of action of Bacillus Calmette-Guerin is adherence to the urothelium with induction of an immunologic and/or inflammatory response . The current study was designed to determine if rabbit bladder mucin removal results in increased Bacillus Calmette-Guerin urothelial adherence . PAS and colloidal iron stains were used to demonstrate that intravesical instillation of 50% acetone renders rabbit bladder urothelium mucin deficient . The urothelium remains mucin deficient at two hours, but by 24 hours the mucin layer has been regenerated . Two hours following intravesical 3H-labeled Escherichia coli administration, bacterial adherence was 29-fold greater in mucin deficient than mucin intact rabbits (p = 0.05) . By 12 hours, the difference in adherence was not significant . Two hours following intravesical administration of 3H-labeled Bacillus Calmette Guerin, mucosal adherence was 21-fold greater in mucin deficient compared to mucin intact rabbits (p = 0.002) . After mucin removal, Bacillus Calmette Guerin urothelial adherence was significantly increased . The significant increase in Bacillus Calmette Guerin adherence after mucin removal may be clinically exploitable. J Infect Dis, 1992 Feb, 165(2), 235 - 44 Epidemiologic classics of Carter, Maxcy, Trudeau, and Smith; Woodward TE; Four great epidemiologists whose work so concisely linked clinical observations, epidemiologic clues, and logical preventive measures are discussed . Henry Rose Carter set the stage for the Walter Reed successes in Cuba by showing that 9-16 days must elapse after contact before yellow fever develops . This provided the link for the Reed group to allow the "virus" to incubate in the mosquito before becoming infectious . Kenneth Maxcy clarified the controversy between endemic typhus fever and Brill's disease in the southeastern United States . His clinical, epidemiologic, and laboratory findings led him to propose that the causative organism (Rickettsia typhi) was in rodents and the probable vector, fleas . When confirmed, effective control measures were applied . Two other American investigators, Edward L . Trudeau and Theobald Smith, helped prove Robert Koch wrong on three counts: (1) Tuberculin is not an effective therapeutic agent for tuberculosis; (2) there are two distinct types of tubercle bacillus, human and bovine; and (3) the bovine form of Mycobacterium tuberculosis is remarkably pathogenic for humans . The significance of these findings is unlimited. Appl Environ Microbiol, 1992 Feb, 58(2), 642 - 6 Insecticidal properties of a crystal protein gene product isolated from Bacillus thuringiensis subsp . kenyae; Masson L et al.; A protoxin gene, localized to a high-molecular-weight plasmid from Bacillus thuringiensis subsp . kenyae, was cloned on a 19-kb BamHI DNA fragment into Escherichia coli . Characterization of the gene revealed it to be a member of the CryIE toxin subclass which has been reported to be as toxic as the CryIC subclass to larvae from Spodoptera exigua in assays with crude E . coli extracts . To directly test the purified recombinant gene product, the gene was subcloned as a 4.8-kb fragment into an expression vector resulting in the overexpression of a 134-kDa protein in the form of phase-bright inclusions in E . coli . Treatment of solubilized inclusion bodies with either trypsin or gut juice from the silkworm Bombyx mori resulted in the appearance of a protease-resistant 65-kDa protein . In force-feeding bioassays, the purified activated protein was highly toxic to larvae of B . mori but not to larvae of Choristoneura fumiferana . In diet bioassays with larvae from S . exigua, the purified protoxin was nontoxic . However, prior activation of the protoxin by tryptic digestion resulted in the appearance of some toxic activity . These results demonstrate that this new subclass of protein toxin may not be useful for the control of Spodoptera species as previously reported . Hierarchical clustering of the nine known lepidopteran-specific CryI toxin subclasses through multiple sequence alignment suggests that the toxins fall into four possible subgroups or clusters. Appl Environ Microbiol, 1992 Feb, 58(2), 525 - 31 Characterization of an extracellular protease inhibitor of Bacillus brevis HPD31 and nucleotide sequence of the corresponding gene; Shiga Y et al.; A novel proteinaceous protease inhibitor was isolated from the culture supernatant of Bacillus brevis HPD31 . The protease inhibitor of B . brevis (designated BbrPI) was produced extracellularly in multiple forms having at least three different molecular weights . One of them, BbrPI-a, was purified to near homogeneity and only showed inhibitory activity toward serine proteases, such as trypsin, chymotrypsin, and subtilisin . BbrPI was presumed to form a trypsin-inhibitor complex in a molar ratio of 1:1 . The inhibitor was found to be heat resistant at neutral and acidic pHs . The gene coding for BbrPI was cloned into Escherichia coli, and its nucleotide sequence was determined . The sequence suggested that BbrPI is produced with a signal peptide of 24 amino acid residues . The amino acid sequence of the protein deduced from the DNA sequence contained the amino acid sequences of amino termini of the inhibitors, a, b, and c, and their putative precursor determined chemically . The molecular weight of the precursor was about 33,000, and the molecular weights of inhibitors a, b, and c were about 22,000, 23,500, and 24,000, respectively . It is presumed that the secreted precursor protein, which is probably inactive, is cleaved by protease into several active protease inhibitor molecules . BbrPI shows no significant homology to the protease inhibitors described previously and is unique in not having any cysteine residues in its molecule. Appl Environ Microbiol, 1992 Feb, 58(2), 520 - 4 Isolation and partial characterization of an 87-kilodalton beta-1,3-glucanase from Bacillus circulans IAM1165; Aono R et al.; Bacillus circulans IAM1165 produces at least two extracellular beta-1,3-glucanases that lyse fungal cell walls . One of these extracellular enzymes was purified to homogeneity . The molecular mass was 87 kDa, and the pI was 4.3 . The optimum temperature of the enzyme reaction was 70 degrees C when laminarin (a soluble beta-1,3-glucan) was used as the substrate . The pH range of the enzyme was broad (pH 4.5 to 9.0), and the optimum pH was 6.5 . The enzyme is an endo beta-1,3-glucanase and has a random cleavage pattern. Appl Environ Microbiol, 1992 Feb, 58(2), 507 - 12 Properties of a 72-kilodalton mosquitocidal protein from Bacillus thuringiensis subsp . morrisoni PG-14 expressed in B . thuringiensis subsp . kurstaki by using the shuttle vector pHT3101; Chang C et al.; The mosquitocidal properties of Bacillus thuringiensis subsp . israelensis and B . thuringiensis subsp . morrisoni PG-14 are attributable to protein inclusions grouped together within a parasporal body . In both of these strains, the mosquitocidal activity resides in proteins with molecular masses of 27, 72, 128, and 135 kDa . In an attempt to determine the toxicity of each protein, the shuttle vector pHT3101 was used to express the cryIVD gene (encoding the 72-kDa CryIVD protein) from B . thuringiensis subsp . morrisoni in an acrystalliferous mutant of B . thuringiensis subsp . kurstaki . With this system, parasporal inclusions of the 72-kDa protein were obtained that were comparable in size, shape, and toxicity to those produced by parental B . thuringiensis subsp . morrisoni . The inclusions were bar shaped, measured 500 by 300 by 150 nm, and were easily visible with phase-contrast microscopy by 16 h of cell growth . A 50% lethal concentration of 64 ng/ml for these inclusions was determined in bioassays against fourth instars of Culex quinquefasciatus, which was similar to the 50% lethal concentration of 55 ng/ml obtained for the 72-kDa inclusion from B . thuringiensis subsp . israelensis . In contrast, expression of the cryIVD gene in Escherichia coli was very low and only detectable by immunoblot analysis . These results demonstrate that the pHT3101-B . thuringiensis expression system can be used to express the CryIVD protein in quantities and with properties comparable to that obtained with the natural host . This system may prove useful for the expression of other B . thuringiensis proteins and, in particular, for reconstitution experiments with inclusions produced by the mosquitocidal subspecies of B . thuringiensis. Clin Investig, 1992 Feb, 70(2), 89 - 98 Endothelial heterogeneity and the acquired immunodeficiency syndrome: a paradigm for the pathogenesis of vascular disorders; Goerdt S et al.; Vascular disorders comprise a wide range of diverse disease entities . Correspondingly, vessels, and even more so the endothelial which line them, show a remarkable extent of heterogeneous differentiation, e.g . between the blood vascular and lymphatic systems, along the length of the vascular trees, and in the microvascular beds of various organs . The most important morphologic criterion to discriminate between endothelia is continuity (continuous endothelial cell layer and well-formed basement membrane) versus discontinuity (intra- or intercellular gaps and/or reduced or missing basement membrane) . Most blood vascular endothelia are of the continuous type, while most sinusoidal and lymphatic endothelia are discontinuous by these criteria . Antigen expression corroborates these morphologic data in that CD31, CD34, and 1F10 antigen are exclusively expressed in continuous endothelia, while MS-1 antigen is preferentially expressed in non-continuous sinusoidal endothelia . In contrast, no specific marker has as yet been described for lymphatic endothelia . Endothelial heterogeneity substantially contributes to the pathogenesis of vascular disorders . For example, in patients with acquired immunodeficiency syndrome the same infectious agent may cause either bacillary angiomatosis (a lobular capillary proliferation) or peliosis (sinusoidal dilatation, endothelial denudation, and development of blood-filled cysts) depending on whether the affected organs have predominantly continuous endothelia or noncontinuous sinusoidal endothelia . Moreover, in Kaposi's sarcoma, it is still an open question of whether the lesion is derived from blood vascular or lymphatic endothelia (Kaposi's sarcoma cells in situ do not express the von Willebrand factor+, PAL-E+, 1F10+ phenotype of mature, resting blood vascular endothelia) . It is also unresolved how endothelia of either type may be differentially induced to dedifferentiate and how they are recruited into the lesion . Clearly, knowledge about endothelial heterogeneity is still too incomplete to identify the actual mechanisms and molecules that govern the pathogenesis of vascular disorders (including still others than those mentioned here such as atherosclerosis, diabetic angiopathy, and rheumatoid arthritis) affecting distinct endothelia . Further efforts in antigenic phenotyping and in cell and molecular biology of heterogeneously differentiated endothelia should be made to improve this state of affairs. Zentralbl Bakteriol, 1992 Feb, 276(3), 303 - 12 Toxic properties of Bacillus cereus strains isolated from different foodstuffs; Hostacka A et al.; Toxic activity of 89 culture filtrates of Bacillus cereus strains were examined . From the total number of the filtrates tested, 3 were found to cause fluid accumulation in the ligated rabbit ileal loop . 71 samples caused a vascular permeability increase in the rabbit skin corresponding to an intermediate or strong toxin production . 59 of the 89 culture filtrates were cytotoxic on Vero cells, 31 had a lethal effect, 81 induced hemolysis of 0.5% rabbit erythrocytes and 55 tested samples showed lecithinase activity . The results showed that relationships exist between the extent of the permeability reaction and cytotoxicity, lecithinase and lethality . No relationship was found between permeability reaction size and reaction in the ligated rabbit ileal loop, nor a strict correlation between permeability reaction and hemolysin . Similarly, the part of culture filtrates showing a weak, or no reaction in the rabbit skin showed hemolytic activity. J Am Acad Dermatol, 1992 Feb, 26(2 Pt 2), 322 - 5 An eruptive vascular proliferation resembling acquired tufted angioma in the recipient of a liver transplant; Chu P et al.; A 59-year-old man developed grouped erythematous papules on the skin of the right axilla and arm shortly after orthotopic liver transplantation . Histologic changes closely resembled those of acquired tufted angioma, with capillary lobules studding the dermis . The lesions spontaneously involuted over several months . Histopathologic and immunohistochemical findings excluded Kaposi's sarcoma and bacillary angiomatosis, the two vascular proliferations most frequently seen in the immunocompromised host . We believe that our case is one of acquired tufted angioma appearing as an eruptive condition similar to eruptive pyogenic granuloma, to which it seems closely related . The involution of lesions in this case is unexplained. Mol Microbiol, 1992 Feb, 6(4), 529 - 46 Four homologous domains in the primary structure of GrsB are related to domains in a superfamily of adenylate-forming enzymes; Turgay K et al.; The entire nucleotide sequence of the Bacillus brevis grsB gene encoding the gramicidin S synthetase 2, which activates and condenses the four amino acids proline, valine, ornithine and leucine has been determined . The gene contains an open reading frame of 13,359 bp which encodes a protein of 4453 amino acids with a predicted Mr of 510,287 . The gene is located within the gramicidin S biosynthetic operon, also containing the genes grsT and grsA, whose nucleotide sequences have been determined previously . Within the GrsB amino acid sequence four conserved and repeated domains of about 600 amino acids (45-50% identity) have been identified . The four domains are separated by non-homologous sequences of about 500 amino acids . The domains also share a high degree of similarity (20-70%) with eight peptide synthetases of bacterial and fungal origin as well as with conserved sequences of nine other adenylate-forming enzymes of diverse origin . On the basis of sequence homology and functional similarities, we infer that those enzymes share a common evolutionary origin and present a phylogenetic tree for this superfamily of domain-bearing enzymes. Nippon Ganka Gakkai Zasshi, 1992 Feb, 96(2), 271 - 5 {An HLA-B27 negative case of ankylosing spondylitis with acute iridocyclitis}; Yoshikawa M et al.; The author experienced a 34-year-old male case of acute iridocyclitis with fibrinous exudates accompanied by ankylosing spondylitis with negative HLA-B27 . Who was diagnosed definite ankylosing spondylitis by the New York criteria . The cause of ankylosing spondylitis was postulated to be a cross reaction between HLA-B27 and a certain Gram-negative bacillus . However, our case with negative HLA-B27 suggested the causes could comprise multiple factors in addition to HLA-B27 . Since the clinical picture of iridocyclitis observed in our case showed no difference with those of HLA-B27 positive cases, iridocyclitis associated with ankylosing spondylitis is considered to be the same entity regardless of HLA-B27 being present or not. Hematol Oncol Clin North Am, 1992 Feb, 6(1), 117 - 27 Intravesical therapy; Herr HW; Intravesical therapy is given as an adjunct to transurethral resection of superficial bladder tumors to eradicate existing disease, inhibit tumor recurrence, and prevent progression to muscle invasion or metastasis . Of the available agents, bacillus Calmette-Guerin (BCG) fulfills these goals and appears superior to chemotherapeutic drugs. Gene, 1992 Feb 1, 111(1), 141 - 2 Bst71I: an isoschizomer of the type-IIS restriction enzyme, BbvI, recognizing the GCAGC(8/12) site; Schoenfeld T et al.; A new type-IIS restriction enzyme, Bst71I, with the specificity 5'-GCAGC(N)8/3'-CGTCG(N)12 was isolated from Bacillus stearothermophilus (Promega No . 71) . This enzyme is an isoschizomer of BbvI with somewhat improved characteristics for use by molecular biologists. EMBO J, 1992 Feb, 11(2), 785 - 92 Cloning of the complete biosynthetic gene cluster for an aminonucleoside antibiotic, puromycin, and its regulated expression in heterologous hosts; Lacalle RA et al.; Puromycin, produced by Streptomyces alboniger, is a member of the large group of aminonucleoside antibiotics . The genes pac and dmpM, encoding a puromycin N-acetyl transferase and an O-demethyl puromycin O-methyltransferase, respectively, are tightly linked in the DNA of S . alboniger . The entire set of genes encoding the puromycin biosynthesis pathway was cloned by screening a gene library from S . alboniger, raised in the low copy number cosmid pKC505, with a DNA fragment containing pac and dmpM . Puromycin was identified by biochemical and physicochemical methods, including 1H NMR, in the producing transformants . This pathway was located in a single DNA fragment of 15 kb which included the resistance, structural and regulatory genes and was expressed when introduced into two heterologous hosts Streptomyces lividans and Streptomyces griseofuscus . In addition to pac and dmpM, two other genes have been identified in the pur cluster: pacHY, which determines an N-acetylpuromycin hydrolase and prg1, whose deduced amino acid sequence is significantly similar to that of degT, a Bacillus stearothermophilus pleiotropic regulatory gene. Br J Urol, 1992 Feb, 69(2), 147 - 50 Intravesical BCG treatment for superficial bladder cancer: long-term results using two different strains of BCG; Mukherjee A et al.; A total of 21 patients received intravesical BCG for the prevention of recurrence of superficial transitional cell carcinoma of the bladder . Twelve patients received the Glaxo strain and 9 the Pasteur strain of the bacillus . Although thought to have different anti-tumour activities owing to a difference in antigenicity, these 2 strains produced similar tumour-free rates at 5 years' mean follow-up with an overall tumour-free rate of 43%. Biotechnol Appl Biochem, 1992 Feb, 15(1), 48 - 58 Cyclomaltodextrin glucanotransferase from Bacillus circulans E 192 . I . Purification and characterization of the enzyme; Bovetto LJ et al.; The cyclomaltrodextrin glucanotransferase (CGTase) {1,4-alpha-D-glucan:4-alpha-D-(1,4-alpha-D-glucano)-transferase (cyclizing), EC 2.4.1.19} from Bacillus circulans E 192 has been purified to homogeneity by Cetavlon treatment, ammonium sulfate precipitation, DEAE Trisacryl M chromatography, Q Fast Flow chromatography, and affinity on beta-cyclodextrin-Sepharose 4B . Two isoenzymes were separated by FPLC on a Mono Q column . Their isoelectric points were estimated as 6.7 and 6.9 and they represented 13 and 87%, respectively, of the initial activity . Their molecular weight, pH, and temperature optima were estimated as 78,000, 5.5, and 60 degrees C, respectively . Kinetic parameters indicated that both enzymes had the same properties; they preferentially modified high-molecular-weight substrates to produce cyclodextrins . The apparent Vmax and Km values for soluble starch were 43 mumol of beta-cyclodextrin/min/mg of protein and 0.57% (w/v), respectively . Although this CGTase is not markedly thermostable, it is protected against heat denaturation by substrate, product, and/or calcium ions . The ratios of alpha-, beta-, and gamma-cyclodextrins produced have been determined as 1/7/2 in the initial phase of the reaction and 3/3/1 at equilibrium. Can J Microbiol, 1992 Feb, 38(2), 153 - 6 Purification of an enterotoxin produced by Bacillus cereus by immunoaffinity chromatography using a monoclonal antibody; Shinagawa K et al.; A murine monoclonal antibody (MAb) with high reactivity to an enterotoxin produced by Bacillus cereus was used to prepare an immunoadsorbent for purification of the enterotoxin . By immunoaffinity chromatography using the immunoadsorbent, approximately 25% of crude enterotoxin applied was recovered in the eluate . The purified enterotoxin was found to be electrophoretically and antigenically homogeneous . It also showed vascular permeability activity and mouse lethality, and caused fluid accumulation in mouse ligated intestinal loops, whereas it did not show any hemolytic and lecithinase activities . Thus, immunoaffinity chromatography proved useful in the purification of enterotoxin produced by B . cereus in terms of recovery, purity, and relative ease of performing the purification. Cent Afr J Med, 1992 Feb, 38(2), 62 - 6 Immunisation and nutritional status of under-fives in rural Zambia; Zondag AM et al.; PIP: In 1987 the nutritional status of Zambian children under 5 years of age was studied in 3 regions around Kamoto Hospital with the objective of exploring the prevalence if malnutrition and contributing factors such as maternal education and immunization status . Jumbe was within easy reach of the hospital with a relatively high standard of living . Masumba and Kakumbi were different areas in one region with their own health center further away from the hospital . Chibembe was isolated without good roads . The nutritional status of 1-5 year old children was measured by the Mid Upper Arm Circumference (MUAC) . A questionnaire with 22 questions queried mothers about education, breast feeding, meals, water supply, and sanitation . A total of 1251 children were observed, 1222 under age 5, and 29 a little older . 40% of mothers had no education and 54% had some primary education (15.2% passed grade 4, 7.3% reached grade 6, and 18.2% finished grade 7) . Less than 5% attended secondary school, and only 1% of mothers finished it . In Chibembe almost 50% of mothers had no education, secondary school education was the lowest of the regions, while in Jumbe was the highest . Immunizations included Bacillus Calmette-Guerin (BCG) at birth, diphtheria-tetanus-pertussis (DTP I, II, III, and a booster), oral polio vaccine (OPV) I, II, III, and a booster, and measles . The Chibembe region has the highest number of incomplete immunizations