Am J Prev Med, 2002 Feb, 22(2), 117 - 9 Screening for tuberculosis among orphans in a developing country; Francis J et al.; BACKGROUND: There has been a recent resurgence of tuberculosis (TB) as a worldwide public health concern . It is a well-documented health problem in Haiti, where the reported prevalence is 123.9 per 100,000 population . While the Centers for Disease Control and Prevention (CDC) and World Health Organization (WHO) policy papers for the eradication of TB recommend screening in shelters, prisons, and other congregate-living facilities, screening in institutional orphanages is not specifically mentioned . METHODS: A total of 445 orphans with no documentation of Bacillus Calmette-Guerin (BCG) vaccine at six orphanages representing urban, suburban, and rural Haiti were screened for TB using standardized purified protein derivative (PPD) . An inoculum of 0.1 cc was introduced intradermally, with site checks for induration at 48-72 hours after inoculation . Induration >5 mm was recorded as positive . Active cases of TB were defined by symptoms such as fever, night sweats, cough greater than 1 month, or signs of extrapulmonary TB (like scrofula or Pott's disease) . RESULTS: The number of positive PPD tests was 170 of the 445 children tested, or 38,202 per 100,000; there were 10 active cases . The overall prevalence of tuberculin positivity in the three orphanages with one or more active cases was 44%, compared to 13% in the three with no identified active cases . CONCLUSIONS: The data indicate that TB prevalence among the orphans screened was 1123 per 100,000 . This is substantially greater than that reported for the general Haitian population . These data thus suggest children living in orphanages as a previously unidentified high-risk group for TB infection . Further investigation of TB is recommended for orphans in developing countries. Aviakosm Ekolog Med, 2000, 34(5), 42 - 5 {Effects of heavy charged particles on strains of microorganisms-producers of biologically active substances}; Kanaeva EN et al.; Effects of heavy charged particles on strains of Bacillus thuringiensis ssp . Kurstaki Z-52 and Arthrobacter OC-1 have been studied . Evidence was obtained that heavy charged particles impact the morphologocultural and physiological properties of culture . As noted, the conditions of orbital flight may be considered a source of mutagenic effects on cultures of microorganisms. Ocul Immunol Inflamm, 2001 Sep, 9(3), 207 - 10 BCG inoculation and corticosteroid sensitivity; Tanaka T et al.; PURPOSE: This study was conducted to clarify the relationship between Bacille de Calmette-Guerin (BCG) inoculation and corticosteroid sensitivity . METHODS: Lewis and WKAH rats were inoculated with BCG vaccine weekly beginning at six weeks of age . At 15 weeks, spleen cells were cultured with PHA and betamethasone for 24 hours . RNA was extracted and interleukin-2 gene expression was measured using a real-time PCR method . RESULTS: BCG inoculation sensitized Lewis rats, but not WKAH rats, to PPD stimulation . In Lewis rats, IL-2 gene expression increased after BCG inoculation, while betamethasone suppressed IL-2 expression . In WKAH rats, the expression of IL-2 was not affected by repeated BCG inoculations and betamethasone had no effect on BCG-treated rats . CONCLUSION: BCG inoculation may affect steroid sensitivity in some strains of rats. Curr Microbiol, 2002 Feb, 44(2), 102 - 5 mbgA-dependent lactose utilization by Bacillus megaterium; Shaw GC et al.; The beta-galactosidase-encoding mbgA gene has recently been cloned from Bacillus megaterium . We now report that disruption of the chromosomal mbgA rendered B . megaterium unable to utilize lactose as a sole carbon source . Complementation of the mbgA mutant with a multicopy plasmid carrying intact mbga restored the ability to utilize lactose for cell growth . Crude extracts from the wild-type B . megaterium cells grown in the presence of lactose exhibited a significant level of lactose-hydrolyzing activity, whereas no activity was observed in crude extracts of the mbgA mutant grown under the same condition . The mbgA gene could also confer the ability of lactose utilization on a lacZ deletion mutant of Escherichia coli . Lactose-hydrolyzing activity was also observed in crude extracts of the lacZ deletion mutant carrying mbgA on a multicopy plasmid . In addition, inactivation of the chromosomal mbgA did not affect lactose induction of expression of the mbgA promoter-xylE transcriptional fusion . Taken together, these results suggest that mbgA is essential for lactose utilization by B . megaterium, but is not involved in generation of the intracellular inducer for lactose induction of mbgA expression. Curr Microbiol, 2002 Feb, 44(2), 81 - 7 Physical mapping of the Bacillus thuringiensis subsp . kurstaki and alesti chromosomes; Lovgren A et al.; Two strains of the well-known insect pathogen and biopesticide, Bacillus thuringiensis (Bt), belonging to subspecies alesti (strain Bt5) and kurstaki (strain Bt213), were chosen for genetic characterization . The two strains belong to different serotypes and are currently classified into different subspecies, although their insecticidal activity is similar . Physical maps were constructed of Bt alesti and Bt kurstaki using Pulsed Field Gel Electrophoreses (PFGE), and the map positions of several genes were determined . The 5.5 Mb combined genetic and physical chromosome maps of the two strains were found to be indistinguishable, and the only differences detected between the strains were of extrachromosomal origin . A cryIA toxin gene probe hybridised to a chromosome fragment and to two extrachromosomal elements in both strains, migrating as 100 kb and 350 kb, respectively . In addition a cry hybridizing extrachromosomal element migrating as 80 kb was present only in Bt alesti . Both strains were also found to contain sequences hybridizing to an enterotoxin (hbla) gene probe . Such sequences were positioned on the 350 kb extrachromosomal element, as well as on the chromosome. Cell Mol Life Sci, 2001 Dec, 58(14), 2136 - 43 Substrate binding and catalytic mechanism of class B beta-lactamases: a molecular modelling study; Prosperi-Meys C et al.; Increased resistance to beta-lactam antibiotics is mainly due to beta-lactamases whose production by pathogenic bacteria makes their broad activity spectrum especially frightening . X-ray structures of several zinc beta-lactamases have revealed the coordination of the two metal ions, but their mode of action remains unclear . Geometry optimisation of stable complexes along the reaction pathway of benzylpenicillin hydrolysis highlighted a proton shuttle occurring from D120 of the Bacillus cereus beta-lactamase to the beta-lactam nitrogen via Zn2 which is central to the network . First, the Zn1 ion has a structural role maintaining Zn-bound waters, WAT1 and WAT2, either directly or through the Zn1 tetrahedrally coordinated histidine ligands . The Zn2 ion has a more catalytic role, stabilising the tetrahedral intermediate, accepting the beta-lactam nitrogen atom as a ligand . The role of Zn2 and the flexibility in the coordination geometry of both Zn ions is of crucial importance for catalysis. J Vector Ecol, 2001 Dec, 26(2), 221 - 31 Mosquito larval control with Bacillus sphaericus: reduction in adult populations in low-income communities in Nonthaburi Province, Thailand; Mulla MS et al.; During 1999 and 2000 several larvicidal treatments of Bacillus sphaericus strain 2362 water dispersible granular (WDG) formulations were made at 50 to 200 mg/m2 in mosquito developmental sites in low-income communities in Nonthaburi Province, Thailand to determine whether larviciding dense populations would results in a noticeable reduction of adult mosquitoes in small treated areas . In the treated area in 1999 (Soi Jumpa), immature populations were suppressed to extremely low levels for extended periods, especially at the higher dosages . This decline in immature populations was followed by a substantial decline in adult mosquitoes . There was a lag of 7 to 14 days post-larval treatments before maximum decline in adults was noted . Adults that emerged prior to treatments survived for 7-14 days or longer, thus no drastic reduction was noted soon after treatments . Despite a slight resurgence in adult mosquitoes during the middle of the experimental period, adult female mosquitoes (over 98% Cx quinquefasciatus), remained low during the 5-month period of trials . During the last 2 weeks (17 days post last treatment) of the experimental period, female populations reached the pre-treatment level . During the 2000 tests at Wat Pikul reduction in larvae was 87-98% for 7 weeks after first treatment at 200 mg/m2, resulting in a reduction of 24 to 73% (2 and 7 days post-treatment respectively) and 87 to 98 (2-6 weeks) in the adults . In the second and third treatments at 50 mg/m2, larval control and subsequent adult reduction were lower and shorter-lived than at the high dosage, and the fourth treatment at 100 mg/m2 did not yield a high level of reduction in the larvae (18 to 33%), but reduction of adults was still 80% . The final fifth treatment at 200 mg/m2 yielded only 18% control of larvae, suggesting tolerance to B . sphaericus at this site . It was shown that at both treated sites repeated treatments with a larvicide such as B . sphaericus could result in substantial reduction in adult mosquitoes . Vigilance for detection of resistance development should be practiced, as resistance could emerge in certain populations following a few treatments. Ying Yong Sheng Tai Xue Bao, 2001 Feb, 12(1), 59 - 62 {Ecological characteristics of phytoplankton in waters of biological-controlling and ordinary rice fields}; Liu W et al.; Through investigations from March to June 1998 on the phytoplankton in waters of biological-controlled and ordinary rice fields, 112 species of phytoplankton were found, of which, Bacillariophyta, Euglenophyta and Chlorophyta were dominant . The comparison of the species in two rice fields showed that in biological-controlled rice field, there were more species of Euglenophyta, with 5 most dominant species, i.e., 2 of Euglenophyta, 2 of Bacillariophyta and 1 of Chlorophyta . In ordinary rice field, there were more Bacillariophyta species, in which, 5 most dominant species belonged to Bacillariophyta, except Scenedesmus bijuga to Chlorophyta . The biodiversities of phytoplankton and their evennesses were also analyzed with the period from the seedlings being planted to the rice fields being dried, showing that a little increase in their biodiversity mainly caused by the increase of species number and an evident decrease in their evennesses. J Biol Chem, 2002 Apr 5, 277(14), 11941 - 8 Epub 2002 Jan 25. The ADP-ribosylating mosquitocidal toxin from Bacillus sphaericus: proteolytic activation, enzyme activity, and cytotoxic effects; Schirmer J et al.; The mosquitocidal toxin (MTX) from Bacillus sphaericus SSII-1 is a approximately 97-kDa protein sharing sequence homology within the N terminus with the catalytic domains of various bacterial ADP-ribosyltransferases . Here we studied the proteolytic activation of the ADP-ribosyltransferase activity of MTX . Chymotrypsin treatment of the 97-kDa MTX holotoxin (MTX(30-870)) results in a 70-kDa putative binding component (MTX(265-870)) and a 27-kDa enzyme component (MTX(30-264)), possessing ADP-ribosyltransferase activity . Chymotryptic cleavage of an N-terminal 32-kDa fragment of MTX (MTX(30-308)) also yields MTX(30-264), but the resulting ADP-ribosyltransferase activity is much greater than that of the processed MTX(30-870) . Kinetic studies revealed a K(m) NAD value of 45 microm for the processed 32-kDa MTX fragment, and a K(m) NAD value of 1300 microm for the processed holotoxin . Moreover, the k(cat) value for the activated MTX(30-308) fragment was about 10-fold higher than that for the activated holotoxin (MTX(30-870)) . Precipitation analysis showed that the 70-kDa proteolytic fragment of MTX remains noncovalently bound to the N-terminal 27-kDa fragment, thereby inhibiting ADP-ribosyltransferase and NAD glycohydrolase activities . Glu(197) of MTX(30-264) was identified as the "catalytic" glutamate that is conserved in all ADP-ribosyltransferases . Whereas mutated MTX(30-264)E197Q has neither ADP-ribosyltransferase nor NAD glycohydrolase activity, mutated MTX(30-264)E195Q possesses glycohydrolase activity but not transferase activity . Transfection of HeLa cells with a vector encoding a fusion protein of MTX(30-264) with a green fluorescent protein led to cytotoxic effects characterized by cell rounding and formation of filopodia-like protrusions . These cytotoxic effects were not observed with the catalytically inactive MTX(30-264)E197Q mutant, indicating that the MTX enzyme activity is essential for the cytotoxicity in mammalian cells. Protein Expr Purif, 2002 Feb, 24(1), 71 - 5 Biochemical analysis of a native and proteolytic fragment of a high-molecular-weight thermostable lipase from a mesophilic Bacillus sp; Dosanjh NS et al.; An extracellular lipase was isolated from the cell-free broth of Bacillus sp . GK 8 . The enzyme was purified to 53-fold with a specific activity of 75.7 U mg(-1) of protein and a yield of 31% activity . The apparent molecular mass of the monomeric protein was 108 kDa as estimated by molecular sieving and 112 kDa by SDS-PAGE . The proteolysis of the native molecule yields a low molecular weight component of 11.5 kDa that still retains the active site . It was stable at the pH range of 7.0-10.0 with optimum pH 8.0 . The enzyme was stable at 50 degrees C for 1 h with a half life of 2 h, 40 min, and 18 min at 60, 65, and 70 degrees C, respectively . With p-nitrophenyl laurate as substrate the enzyme exhibited a K(m) and V(max) of 3.63 mM and 0.26 microM/min/ml, respectively . Activity was stimulated by Mg(2+) (10 mM), Ba(2+) (10 mM), and SDS (0.1 mM), but inhibited by EDTA (10 mM), phenylmethane sulfonyl fluoride (100 mM), diethylphenylcarbonate (10 mM), and eserine (10 mM) . It hydrolyzes triolein at all positions . The fatty acid specificity of lipase is broad with little preference for C(4) and C(18:1) . Thermostability of the proteolytic fragment at 60 degrees C was observed to be 37% of the native protein . The native enzyme was completely stable in ethylene glycol and glycerol (30% v/v each) for 60 min at 65 degrees C . J Invertebr Pathol, 2001 Oct, 78(3), 160 - 9 Coexpression of chitinase and the cry11Aa1 toxin genes in Bacillus thuringiensis serovar israelensis; Sirichotpakorn N et al.; At the spore stage, a cloned chitinase gene was coexpressed with the regulatory gene p19 and the toxin gene cry11Aa1 in the hosts Bacillus thuringiensis serovar israelensis strains 4Q2-72 and c4Q2-72 . The chitinase gene was derived from a high-chitinase producer, Bacillus licheniformis TP-1 . Two transcriptional fusion plasmids between the p19 or p19-cry11Aa1 genes and the promoterless chitinase gene were constructed . In transcription order, the p16-19CHI construct contained the p19 gene together with the chitinase gene only while the p16-1968CHI construct contained p19 together with the toxin gene cry11Aa1 and the chitinase gene . The inserted sequences were regulated by a spore-specific promoter located upstream of p19 . The recombinant chitinase of all transformed B . thuringiensis serovar israelensis strains was initially synthesized at low level at about 9 h of growth when a portion of the cells started to sporulate . It increased thereafter and reached maximum levels of 5.5, 4.9, and 4.7 mU/ml at 48 h, for strain 4Q2-72 transformed with p16-19CHI and p16-1968CHI and strain c4Q2-72 transformed with p16-19CHI, respectively . This activity was approximately 2 times higher than the maximum activity (2.7 mU/ml) of the parental strain, B . licheniformis TP-1 . Although crude chitinase alone from B . thuringiensis serovar israelensis c4Q2-72 (p16-19CHI) at 4.5 mU/ml caused 40% mortality in second instar Aedes aegypti larvae, transformants containing the chitinase alone or in combination with cry11Aa1 resulted in lower toxicity to A . aegypti larvae than the untransformed 4Q2-72 host . For example the LC(50) for the transformed 4Q2-72 harboring the chitinase gene only (p16-19CHI) was 5.6 x 10(4) +/- 0.7 x 10(4) cells, 40 times higher than that of the untransformed host at 1.4 x 10(3) +/- 0.19 x 10(3) . The lower toxicity correlated with poor sporulation in the transformants (i.e., 35 times lower than that in the untransformed host) . However, the transformed 4Q2-72 strain expressing both the chitinase and the cry11Aa1 toxin genes (p16-1968CHI) were only 4-fold less toxic (LC(50) = 5.6 x 10(3) +/- 1.99 x 10(3)) than the untransformed 4Q2-72 hosts even though their spore count was 300 times lower . Since coapplication of crude chitinase from the cloned gene in recombinant strain (c4Q2-72 harboring p16-19CHI) with cell suspensions of B . thuringiensis serovar israelensis 4Q2-72 and its transformants could enhance 3- to 50-fold larvicidal activity, improvement in sporulation ability of these genetically engineered strains and cocrystallization of chitinase with crystal toxins may increase their potential for future insect control . (c)2001 Elsevier Science (USA). Int Immunopharmacol, 2002 Feb, 2(2-3), 325 - 32 Maternal immune stimulation in mice decreases fetal malformations caused by teratogens; Holladay SD et al.; For unknown reasons, non-specific stimulation of the maternal immune system in pregnant mice has what appears to be a broad-spectrum efficacy for reducing birth defects . Immune stimulation by diverse procedures has proven effective, including footpad injection with Freund's complete adjuvant (FCA), intraperitoneal (IP) injection with inert particles to activate resident macrophages, IP injection with attenuated Bacillus Calmette-Guerin (BCG), and intrauterine injection with allogeneic or zenogeneic lymphocytes . Morphologic lesions that were significantly reduced included cleft palate and associated craniofacial defects, digit and limb defects, tail malformations, and neural tube defect (NTD) . Teratogenic stimuli to induce these lesions included chemical agents (2,3,7,8-tetrachlorodibenzo-p-dioxin {TCDD}, ethyl carbamate {urethane}, methylnitrosourea {MNU}, cyclophosphamide {CP}, and valproic acid {VA}), physical agents (X-rays, hyperthermia), and streptozocin (STZ)-induced diabetes mellitus . Limited information is available regarding mechanisms by which such immune stimulation reduced fetal dysmorphogenesis . The collective literature suggests the possibility that immunoregulatory cytokines of maternal origin may be the effector molecules in this phenomenon. Science, 2002 Jan 25, 295(5555), 674 - 6 Plant biotechnology in China; Huang J et al.; A survey of China's plant biotechnologists shows that China is developing the largest plant biotechnology capacity outside of North America . The list of genetically modified plant technologies in trials, including rice, wheat, potatoes, and peanuts, is impressive and differs from those being worked on in other countries . Poor farmers in China are cultivating more area of genetically modified plants than are small farmers in any other developing country . A survey of agricultural producers in China demonstrates that Bacillus thuringiensis cotton adoption increases production efficiency and improves farmer health. J Exp Bot, 2002 Feb, 53(367), 323 - 31 Plant allocation to defensive compounds: interactions between elevated CO(2) and nitrogen in transgenic cotton plants; Coviella CE et al.; Plant allocation to defensive compounds in response to growth in elevated atmospheric CO(2) in combination with two levels of nitrogen was examined . The aim was to discover if allocation patterns of transgenic plants containing genes for defensive chemicals which had not evolved in the species would respond as predicted by the Carbon Nutrient Balance (CNB) hypothesis . Cotton plants (Gossypium hirsutum L.) were sown inside 12 environmental chambers . Six of them were maintained at an elevated CO(2) level of 900 micromol mol(-1) and the other six at the current level of approximately 370 micromol mol(-1) . Half the plants in each chamber were from a transgenic line producing Bacillus thuringiensis (Bt) toxin and the others were from a near isogenic line without the Bt gene . The allocation to total phenolics, condensed tannins, and gossypol and related terpenoid aldehydes was measured . All the treatments were bioassayed against a non-target insect herbivore found on cotton, Spodoptera exigua (Hubner) (Lepidoptera: Noctuidae) . Plants had lower N concentrations and higher C:N ratios when grown in elevated CO(2) . Carbon defensive compounds increased in elevated CO(2), low N availability or both . The increase in these compounds in elevated CO(2) and low N, adversely affected growth and survival of S . exigua . The production of the nitrogen-based toxin was affected by an interaction between CO(2) and N; elevated CO(2) decreased N allocation to Bt, but the reduction was largely alleviated by the addition of nitrogen . The CNB hypothesis accurately predicted only some of the results, and may require revision . These data indicate that for the future expected elevated CO(2) concentrations, plant allocation to defensive compounds will be affected enough to impact plant-herbivore interactions. Rev Biol Trop, 2001 Mar, 49(1), 191 - 7 Diet seasonality and food overlap among fishes of the upper Orituco stream, northern Venezuela; Ortaz M; The diets of four diurnal fish species (Creagrutus bolivari, Knodus deuterodonoides, Knodus sp . and Poecilia reticulata) were examined during a year in the Orituco stream at northern Venezuela . The fishes were sampled monthly from February 1991 to March 1992 (except October 1991 and February 1992) in the stream main channel with a beach seine and a cast net . Diet is reported as frequency of occurrence and numeric proportion because variation in prey sizes was small . Non-parametric statistical tests were applied . A total of 18 distinct prey items were found in stomachs . The diet of these fishes consisted of aquatic insects (Coleoptera, Diptera, Hemiptera, Odonata, Plecoptera and Trichoptera), allochthonous plant matter (fragments of leaves and seeds), microalgae (Chlorophyta and Bacillariophyceae) and terrestrial arthropods (Coleoptera, Diptera, Hymenoptera and Arachnida) . The low percentage of empty guts and the high fullness percentage of guts suggest that food was always abundant . Aquatic insects were more important in the dry season (November-April) while plant matter and terrestrial arthropods increased in the wet season (May-October) . The Proportional Similarity Measure (PS) was high between characid species and low between characids and P . reticulata in the dry season . PS decreased during the wet season because of a reduction in aquatic insect consumption . The seasonal diet shift indicated the greater importance of allochthonous food only in the wet season. Insect Biochem Mol Biol, 2002 Mar 1, 32(3), 255 - 63 Characterization of a novel insect digestive DNase with a highly alkaline pH optimum; Schernthaner JP et al.; A novel DNase from the digestive tract of the spruce budworm (Choristoneura fumiferana) has been isolated and characterized . This DNase has two features that distinguish it from other known DNases: (1) it has a pH optimum of 10.5 to 11; (2) it plays an important role in the conversion of the insecticidal crystal protein from Bacillus thuringiensis to the active DNA-free toxin in the larval gut . Only one digestive DNase with an apparent molecular mass of 23 kDa was found and no associated carbohydrate was detected . It has some similarities to pancreatic DNase I in that divalent alkaline metal ion is required for activity and it is inhibited by monovalent cations . In particular, Mg(2+) and Ca(2+) were the most effective activators . Transition metal ions also activated the enzyme but were less effective . The enzyme is an endonuclease that hydrolyzes single and double stranded DNA but shows a higher specificity for single stranded DNA . The purified enzyme acted synergistically with proteases on crystals from Bacillus thuringiensis to yield the DNA-free toxin . To our knowledge, this is the first characterization of DNase activity in insect larvae and provides strong evidence that a DNase is an integral component of the larval digestive system. J Dermatol, 2001 Dec, 28(12), 762 - 4 Recurrent lupus vulgaris following repeated BCG (Bacillus Calmette Guèrin) vaccination; Sasmaz R et al.; We report a case of lupus vulgaris following BCG (Bacillus Calmette Guerin) vaccination on the right shoulder . The patient had a history of lupus vulgaris on his left shoulder that developed following BCG vaccination and was treated successfully eight years ago. Vaccine, 2002 Jan 15, 20(7-8), 1126 - 33 Influence of sensitisation to environmental mycobacteria on subsequent vaccination against bovine tuberculosis; Buddle BM et al.; Bacille Calmette-Guerin (BCG) is the world's most widely used vaccine, but there are concerns that it provides little protection against pulmonary tuberculosis of humans in countries that have a high prevalence of environmental mycobacteria . Experiments in cattle provide a model to investigate this situation and to develop an improved tuberculosis vaccine . In the third of a series of BCG vaccination trials, calves had high interferon-gamma (IFN-gamma) responses to purified protein derivative (PPD) from Mycobacterium avium prior to vaccination, indicating that infection with environmental mycobacteria had occurred . The calves vaccinated with BCG had minimal protection against an experimental intratracheal challenge with virulent Mycobacterium bovis . In comparison, calves vaccinated with either of two newly-derived attenuated M . bovis strains had significantly better but not complete protection against the development of tuberculous lesions compared to both BCG-vaccinated and non-vaccinated animals . Vaccination with the newly-derived attenuated M . bovis strains induced strong IFN-gamma and interleukin-2 (IL-2) responses to PPD from M . bovis at 2 weeks after vaccination, while BCG vaccination induced only a weak response at this time . In association with the previous two trials, the results suggest that sensitisation of the calves to environmental mycobacteria adversely affected subsequent protective efficacy of BCG . However, the results of vaccination with the other two attenuated M . bovis strains indicated that improved tuberculosis vaccines could be developed for such sensitised animals. Biochemistry, 2002 Jan 29, 41(4), 1398 - 408 PI-specific phospholipase C cleavage of a reconstituted GPI-anchored protein: modulation by the lipid bilayer; Lehto MT et al.; Release of glycosylphosphatidylinositol- (GPI-) anchored ectoenzymes from the membrane by phosphatidylinositol- (PI-) specific phospholipases may play an important role in modulating the surface expression and function of this group of proteins . To investigate how the properties of the host membrane affect anchor cleavage, porcine lymphocyte ecto-5'-nucleotidase (5'-NTase; EC 3.1.3.5) was purified, reconstituted into lipid bilayer vesicles of various lipids, and cleaved using PI-PLC from Bacillus thuringiensis (Bt-PI-PLC) . Bt-PI-PLC activity was highly dependent on the chain length and unsaturation of the constituent phospholipids . Very high rates of cleavage were observed in fluid lipids with a low phase transition temperature (T(m)), in lymphocyte plasma membrane, and in a lipid mixture that formed rafts . Arrhenius plots of the rate of anchor cleavage in various lipids showed a characteristic break at the bilayer T(m), together with a discontinuity close to T(m) . The activation energy for GPI anchor cleavage was substantially higher in gel phase bilayers compared to those in the liquid crystalline phase . The addition of cholesterol simultaneously abolished the phase transition and the large difference in cleavage rates observed above and below T(m) . Inclusion of GM(1) and GT(1b) (components of lipid rafts) in the bilayer reduced the overall activity, but the pattern of the Arrhenius plots remained unchanged . Both gangliosides had similar effects, suggesting that bilayer surface charge has little influence on PI-PLC activity . Taken together, these results suggest that lipid fluidity and packing are the most important modulators of Bt-PI-PLC activity on GPI anchors. J Formos Med Assoc, 2001 Dec, 100(12), 829 - 31 Childhood tuberculosis presenting as an anterior chest wall abscess; Huang CY et al.; Chest wall abscess is a rare manifestation of childhood tuberculosis . We report a case of a tuberculous chest wall abscess in a 4-year-old healthy girl who had received Bacillus Calmette-Guerin (BCG) vaccination at birth . She developed a localized anterior chest wall mass, which was initially mistaken for enchondroma on the chest radiograph . Pathologic examination of the biopsy specimen revealed chronic granulomatous inflammation and positive acid-fast staining, which confirmed the diagnosis of chest wall tuberculosis infection . She received a 12-month course of anti-tuberculous treatment and was perfectly well 1 year later . The chest wall lesion resolved without the need for surgery . In conclusion, tuberculosis should be excluded in children with undiagnosed chest wall lesions, especially in endemic areas, even if they have been vaccinated with BCG . Adequate anti-tuberculosis treatment can result in a complete recovery. FEBS Lett, 2002 Jan 16, 510(3), 201 - 5 Directional degradation of beta-chitin by chitinase A1 revealed by a novel reducing end labelling technique; Imai T et al.; A novel procedure for labelling the molecular ends of beta-chitin crystals has been established . By introducing a hydrazide derivative of biotin at the reducing end of a chitin chain, followed by a specific interaction between biotin and streptavidin coupled with a colloidal gold particle, the chain directionality of beta-chitin microcrystals could be directly visualized by transmission electron microscopy . This method allowed to certify the parallelism of the chitin chains in the beta-chitin microcrystals, and also to label the reducing tips of beta-chitin microcrystals degraded by Bacillus circulans chitinase A1 . With these substrates, the labelling occurred only at their tapered tip, which indicates that the digestion of these crystals proceeded from their reducing end . The generalization of this new labelling method to other polysaccharide crystals is discussed. Zhonghua Nei Ke Za Zhi, 1999 Feb, 38(2), 98 - 100 {The serum bactericidal activity of domestic fleroxacin and lomefloxacin}; Zhao M et al.; OBJECTIVE: To evaluate antibacterial activities in vivo and in vitro of fleroxacin and lomefloxacin . METHODS: Minimal inhibitory concentration (MIC) and serum bactericidal activity (SBA) domestic fleroxacin and lomefloxacin were determined by agar disk dilution method and microplate method respectively . A clinical observation of fleroxacin and lomefloxacin for treating acute bacillary dysentery was carried out . RESULTS: MIC(90) of fleroxacin against E . coli, K . pneumoniae, S . dysenteriae, S . aureus was 0.25-2 mg/L, being the same as that of ofloxacin . However, MIC(90) of lomefloxacin against the above mentioned species was twice or fourfold as that of ofloxacin and fleroxacin . The percent age of peak SBA > or = 1:8 with ofloxacin, fleroxacin and lomefloxacin against E . coli, K . pneumoniae, S . dysenteriae was 80%-100%, 90%-100% and 50%-80% respectively (P < 0.01) . The clinical cure rates of acute bacillary dysentery were 93.3%, 100% and 100% and the bacterial clearance rates were 93.3%, 100% and 100% in the lomefloxacin, fleroxacin and ofloxacin group respectively . CONCLUSION: The antibacterial activity in vivo and in vitro of fleroxacin against E . coli, K . pneumoniae, S . dysenteriae was similar to that of ofloxacin and stronger than that of lomefloxacin. Clin Infect Dis, 2002 Feb 15, 34(4), 447 - 53 Epub 2002 Jan 04. National study of adverse reactions after vaccination with bacille Calmette-Guérin; Turnbull FM et al.; Few large prospective studies of adverse reactions after bacille Calmette-Guerin (BCG) vaccination are available . In a prospective national study of such adverse reactions among 918 subjects (aged 1 day to 54 years) over a 14-month period, 45 vaccinees (5%) reported 53 adverse reactions (23 injection-site abscesses, 14 severe local reactions, 10 cases of lymphadenitis, and 6 other reactions) . Only 1% of vaccinees required medical attention . Reactions, particularly lymphadenitis, were significantly less common in infants <6 months old (but not in subjects aged > or =6 months) vaccinated by trained (vs . untrained) providers (relative risk {RR}, 0.24; 95% confidence interval {CI}, 0.09-0.68) . Injection-site abscesses (RR, 2.96; 95% CI, 1.11-7.90) and severe local reactions (RR, 4.93; 95% CI, 1.11-21.90) were significantly more common in older vaccinees . Local reactions were more frequently reported by adult females than by adult males (RR, 7.18; 95% CI, 1.59-32.45) . Adverse reactions were not significantly associated with any currently available vaccine batch, previous receipt of BCG vaccine, or concomitant administration of other vaccines. Arch Microbiol, 2001 Dec, 177(1), 70 - 80 Epub 2001 Nov 10. Characterization of an S-layer glycoprotein produced in the course of S-layer variation of Bacillus stearothermophilus ATCC 12980 and sequencing and cloning of the sbsD gene encoding the protein moiety; Egelseer EM et al.; The cell surface of Bacillus stearothermophilus ATCC 12980 is completely covered by an oblique lattice which consists of the S-layer protein SbsC . On SDS-polyacrylamide gels, the mature S-layer protein migrates as a single band with an apparent molecular mass of 122 kDa . During cultivation of B . stearothermophilus ATCC 12980 at 67 degrees C instead of 55 degrees C, a variant developed that had a secondary cell wall polymer identical to that of the wild-type strain, but it carried an S-layer glycoprotein that could be separated on SDS-polyacrylamide gels into four bands with apparent molecular masses of 92, 118, 150 and 175 kDa . After deglycosylation, only a single protein band with a molecular mass of 92 kDa remained . The complete nucleotide sequence encoding the protein moiety of this S-layer glycoprotein, termed SbsD, was established by PCR and inverse PCR . The sbsD gene of 2,709 bp is predicted to encode a protein of 96.2 kDa with a 30-amino-acid signal peptide . Within the 807 bp encoding the signal peptide and the N-terminal sequence (amino acids 31-269), different nucleotides for sbsD and sbsC were observed in 46 positions, but 70% of these mutations were silent, thus leading to a level of identity of 95% for the N-terminal parts . The level of identity of the remaining parts of SbsD and SbsC was below 10%, indicating that the lysine-, tyrosine- and arginine-rich N-terminal region in combination with a distinct type of secondary cell wall polymer remained conserved upon S-layer variation . The sbsD sequence encoding the mature S-layer protein cloned into the pET28a vector led to stable expression in Escherichia coli HMS174(DE3) . This is the first example demonstrating that S-layer variation leads to the synthesis of an S-layer glycoprotein. J Biol Chem, 2002 May 3, 277(18), 15293 - 302 Epub 2002 Jan 16. Mycobacterium tuberculosis hemoglobin HbO associates with membranes and stimulates cellular respiration of recombinant Escherichia coli; Pathania R et al.; The truncated hemoglobins HbN and HbO of Mycobacterium tuberculosis H37Rv share little sequence similarity and display structural differences in their EF-loop regions, suggesting distinct function(s) for these hemoglobins . HbO of M . tuberculosis was expressed in Escherichia coli and Mycobacterium smegmatis as a 14.5-kDa homodimeric heme protein exhibiting nearly 50-fold (P(50) approximately 0.51) lower oxygen affinity than HbN . 40-50% of HbO remained associated with the cell membranes and significantly enhanced its respiration in comparison with the membrane fractions of control cells or cells overproducing HbN . Oxygen uptake of HbO-associated membranes was decreased by washing and restored by adding HbO . Additionally, membrane vesicles prepared from terminal oxidase-deficient (cyo(-), cyd(-)) mutants of E . coli did not exhibit significant enhancement in oxygen uptake in the presence of HbO, suggesting its interaction(s) with the electron transport chain . Expression of HbO in Mycobacterium bovis bacillus Calmette-Guerin, an experimental model of M . tuberculosis, was observed (0.2-0.5% of total cellular proteins) throughout its aerobic growth . These results provided evidence for the involvement of HbO with the component of aerobic electron transport chain, suggesting that its function may be related to the facilitation of oxygen transfer during aerobic metabolism of M . tuberculosis . Membrane association properties of HbO may thus play a crucial role in sequestering oxygen and facilitating its availability to internalized M . tuberculosis (an obligate aerobe) under the hypoxic conditions of its intracellular habitat. Infect Immun, 2002 Feb, 70(2), 672 - 8 Failure of the Mycobacterium bovis BCG vaccine: some species of environmental mycobacteria block multiplication of BCG and induction of protective immunity to tuberculosis; Brandt L et al.; The efficacy of Mycobacterium bovis bacillus Calmette-Guerin (BCG) vaccine against pulmonary tuberculosis (TB) varies enormously in different populations . The prevailing hypothesis attributes this variation to interactions between the vaccine and mycobacteria common in the environment, but the precise mechanism has so far not been clarified . Our study demonstrates that prior exposure to live environmental mycobacteria can result in a broad immune response that is recalled rapidly after BCG vaccination and controls the multiplication of the vaccine . In these sensitized mice, BCG elicits only a transient immune response with a low frequency of mycobacterium-specific cells and no protective immunity against TB . In contrast, the efficacy of TB subunit vaccines was unaffected by prior exposure to environmental mycobacteria . Six different isolates from soil and sputum samples from Karonga district in Northern Malawi (a region in which BCG vaccination has no effect against pulmonary TB) were investigated in the mouse model, and two strains of the Mycobacterium avium complex were found to block BCG activity completely. Water Sci Technol, 2001, 44(10), 253 - 9 Effect of surface active agents on the production of biopesticides using wastewater sludge as a raw material; Vidyarthi AS et al.; Effect of surface active agents on the production of Bacillus thuringiensis based biopesticides using secondary wastewater sludge as a raw material was studied . The experiments were conducted in a shake flask and nine different commercial surface active agents were used at a concentration of 0.2% (v/v) . The sludge samples after addition of surface active agents were sterilised at 121 degrees C for 30 min . The progress of biopesticide production was monitored by measuring viable cells and viable spores counts . The entomotoxicity was determined through bioassay against 3rd instar larvae of spruce budworm . ATPLUS 522, Tween 80 and Tween 85 increased the entomotoxicity potential in comparison to the control . The highest increase of 24% in entomotoxicity was observed when sludge was fortified with Tween 80. J Urol, 2002 Feb, 167(2 Pt 1), 494 - 500; discussion 501 Superficial bladder carcinoma treated with bacillus Calmette-Guerin: progression-free and disease specific survival with minimum 10-year followup; Davis JW et al.; PURPOSE: Intravesical bacillus Calmette-Guerin (BCG) treatment of high risk superficial bladder cancer has reduced recurrence and progression, and lengthened disease specific survival . However, documentation of treatment durability is limited . MATERIALS AND METHODS: Between 1981 and 1989, 98 patients with high risk or recurrent transitional cell carcinoma were treated with complete transurethral resection followed by 1 or more 6-week induction courses of BCG, and were followed through 2000 . No maintenance regimen was used . A total of 44 cases were carcinoma in situ plus or minus papillary and 35 were stage T1, which was assigned only if muscularis propria free of tumor was present on the biopsy specimen . RESULTS: Of 98 patients with minimum followup greater than 10 years disease progressed to stage T2 or greater in 27 at a median of 30.7 months (range 1.2 to 143.7), of whom cystectomy was performed in 16, cystectomy for recurrent high risk Ta/T1 disease was required in 10, death from transitional cell carcinoma occurred in 13 at a median of 69.7 months (range 11 to 135), upper tract tumor developed in 13 at a median of 49 months (range 9 to 146) and there was evidence of prostatic urethral involvement in 21 . The 10-year progression-free survival was 67%, and progression-free survival with a retained bladder 59% . The total cystectomy rate was 26%, permitting a number of patients to be rendered disease-free . The 10-year disease specific survival was 85% . CONCLUSIONS: After complete tumor resection and careful pathological staging intravesical BCG for high risk and/or recurrent superficial bladder carcinoma resulted in overall progression-free and disease specific survival rates that support this bladder sparing strategy . Patients must be followed closely and cystectomy recommended for those with an initial incomplete response after initial therapy or recurrent high risk disease. Biosci Biotechnol Biochem, 2001 Nov, 65(11), 2456 - 64 Enzymatic synthesis of sulfated disaccharides using beta-D-galactosidase-catalyzed transglycosylation; Murata T et al.; We have established a unique enzymatic approach for obtaining sulfated disaccharides using Bacillus circulans beta-D-galactosidase-catalyzed 6-sulfo galactosylation . When 4-methyl umbelliferyl 6-sulfo beta-D-galactopyranoside (S6Gal beta-4MU) was used as a donor, the enzyme induced transfer of 6-sulfo galactosyl residue to GlcNAc acceptor . As a result, the desired compound 6'-sulfo N-acetyllactosamine (S6Gal beta1-4GlcNAc) and its positional isomer 6'-sulfo N-acetylisolactosamine (S6Gal beta1-6GlcNAc) were observed by HPAEC-PAD, in 49% total yield based on the donor added, and in a molar ratio of 1:3.5 . With a glucose acceptor, the regioselectivity was substantially changed and S6Gal beta1-2Glc was mainly produced along with beta-(1-1)alpha, beta-(1-3), beta-(1-6) isomers in 74% total yield . When methyl alpha-D-glucopyranoside (Glc alpha-OMe) was an acceptor, the enzyme also formed mainly S6Gal beta1-2Glc alpha-OMe with its beta-(1-6)-linked isomer in 41% total yield based on the donor added . In both cases, it led to the predominant formation of beta-(1-2)-linked disaccharides . In contrast, with the corresponding methyl beta-D-glucopyranoside (Glc beta-OMe) acceptor, S6Gal beta1-3Glc beta-OMe and S6Gal beta1-6Glc beta-OMe were formed in a low total yield of 12% . These results indicate that the regioselectivity and efficiency on the beta-D-galactosidase-mediated transfer reaction significantly depend on the anomeric configuration in the glucosyl acceptors. Biosci Biotechnol Biochem, 2001 Nov, 65(11), 2419 - 27 Binding properties of Bacillus thuringiensis Cry4A toxin to the apical microvilli of larval midgut of Culex pipiens; Yamagiwa M et al.; Cry4A is a dipteran-specific delta-endotoxin produced by Bacillus thuringiensis, and toxic to Culex pipiens (mosquito) larvae . The immunohistochemical staining of the midgut sections of C . pipiens larvae revealed that Cry4A bound in vitro and in vivo to the microvilli of the epithelial cells of posterior midgut and gastric caecae . The binding of digoxigenin-labeled Cry4A (DIG-Cry4A) to the apical microvilli was almost abolished in the presence of excess unlabeled Cry4A, suggesting that the binding of Cry4A to the microvilli was specific . Several Cry4A-specific binding proteins were detected using the ligand blotting technique with DIG-Cry4A . Moreover, an insertion assay was done, where the binding of DIG-Cry4A to the BBMVs was completely irreversible and did not compete with excess unlabeled Cry4A . On the basis of these results, we propose a schematic interpretation for the binding process of Cry4A. Pediatr Infect Dis J, 2002 Jan, 21(1), 49 - 53 Twenty years of pulmonary tuberculosis in children: what has changed? Sanchez-Albisua I, Baquero-Artigao F, Del Castillo F, Borque C, Garcia-Miguel MJ, Vidal ML. OBJECTIVES: To compare the frequency, clinical and radiologic manifestations and source of infection of pulmonary tuberculosis in children treated in our hospital during two decades (1978 through 1987 and 1988 through 1997) and to evaluate the influence of the emergence of HIV infection (since 1985) and the effect of the discontinuation of Calmette-Guerin bacillus (BCG) vaccination (since 1987) on childhood tuberculosis . METHODS: We reviewed 324 children diagnosed with pulmonary tuberculosis in our hospital during the 20 years (1978 through 1997) . The data from 2 decades, 1978 through 1987 and 1988 through 1997, were compared . BCG vaccination in Spain was discontinued in 1987, and HIV infection emerged significantly as a public health problem . RESULTS: An increase in the number of children with single hilar adenopathy was observed (32.2% in 1978 through 1987 vs . 43.4%, in 1988 through 1997, P < 0.05) in comparison with those with parenchymal involvement or a mixed pattern (62.4% in 1978 through 1987 vs . 45.7% in 1988 to 1997) . Frequency in extrapulmonary manifestations in both periods was similar, with a nonsignificant trend toward a lower rate of tuberculous meningitis in the latter decade (10.4 vs . 5.6%, P = 0.07) . We were able to identify an adult source case for 67.1% of the children (100 of 149) in the first decade vs.58.3% (102 of 175) in the second (P = NS); 10.8% of adult contacts but only 2.3% of children (all of them in the second period) were HIV-positive.CONCLUSIONS: Discontinuation of BCG vaccination and emergence of HIV infection have had little influence on childhood tuberculosis in our area. Int J Food Microbiol, 2001 Dec 30, 71(2-3), 131 - 8 Sporicidal action of ozone and hydrogen peroxide: a comparative study; Khadre et al.; Elimination of contaminating spores on packaging materials and food-contact surfaces remains a challenge to the food industry . Hydrogen peroxide and chlorine are the most commonly used sanitizers to eliminate these contaminants, and ozone was recommended recently as an alternative . Hence, we compared the sporicidal action of ozone and hydrogen peroxide against selected foodborne spores of Bacillus spp . Under identical treatment conditions, 11 microg/ml aqueous ozone decreased spore counts by 1.3 to 6.1 log10 cfu/ml depending upon the bacterial species tested . Hydrogen peroxide (10%, w/w), produced only 0.32 to 1.6 log10 cfu/ml reductions in spore counts . Thus, hydrogen peroxide, at approximately 10,000-fold higher concentration, was less effective than ozone against Bacillus spores . Resistance of spores to ozone was highest for Bacillus stearothermophilus and lowest for B . cereus . Therefore, spores of B . stearothermophilus are suitable for testing the efficacy of sanitization by ozone . Electron microscopic study of ozone-treated B . subtilis spores suggests the outer spore coat layers as a probable site of action of ozone. Glycoconj J, 2001 Apr, 18(4), 301 - 8 Linkage of sugar chains to a fragment peptide of FGF-5S by a chemoenzymatic strategy and changes in the rate of proteolytic hydrolysis; Ajisaka K et al.; Various O-linked and N-linked sugar chains were linked enzymatically to a fragment peptide (Leu-Ser-Gln(or Asn)-Val-His-Arg) of FGF-5S . First, galactose was linked with beta-(1-->3)-linkage to GalNAc-linked peptide by a transglycosylation using beta-galactosidase from Bacillus circulans (recombinant) . Then sialic acid was linked with the aid of sialyltransferase from rat liver (recombinant) to give NeuAcalpha-(2-->3)-Galbeta-(1-->3)-GalNAc-linked hexapeptide . Further, a sialylated 2-chain biantennary sugar chain was linked by a transglycosylation using endo N-acetyl-beta-D-glucosaminidase from Mucor hiemalis (endo M, recombinant) . The activity of DNA synthesis in a fibroblast cell line was increased by this glycosylation . The resistance of the obtained glycopeptides towards proteolytic hydrolysis by rat serum and by five proteases was compared with that of original peptide . The resistance was remarkably enhanced by the glycosylation. Nucleic Acids Res, 2002 Jan 15, 30(2), 497 - 506 Contribution of downstream promoter elements to transcriptional regulation of the rice tungro bacilliform virus promoter; He X et al.; Downstream sequences influence activity of the rice tungro bacilliform virus (RTBV) promoter in protoplasts derived from cultured rice cells . We previously identified a DNA element located between positions +50 and +90 relative to the transcription start site to which rice nuclear proteins bind . In this study, using DNA UV crosslinking assays, we show that two rice nuclear proteins bind specifically to this DNA element . We demonstrate that the DNA element enhances RTBV promoter activity in a copy number-dependent manner when transferred to a position upstream of the promoter . In addition, using electrophoretic mobility shift assays, we show that at least two novel nuclear proteins from rice cell suspension cultures bind to a subregion (from +50 to +59) of the DNA element and that a protein from rice root, but not shoot, nuclear extracts interacts with a perfect palindromic sequence motif located within the sequence +45 to +59 . Furthermore, a position-dependent GAGA motif, present in three copies within downstream promoter sequences from +1 to +50, is involved in the regulation of RTBV promoter activity. Bioseparation, 2001, 10(1-3), 99 - 112 High gradient magnetic separation versus expanded bed adsorption: a first principle comparison; Hubbuch JJ et al.; A robust new adsorptive separation technique specifically designed for direct product capture from crude bioprocess feedstreams is introduced and compared with the current bench mark technique, expanded bed adsorption . The method employs product adsorption onto sub-micron sized non-porous superparamagnetic supports followed by rapid separation of the 'loaded' adsorbents from the feedstock using high gradient magnetic separation technology . For the recovery of Savinase from a cell-free Bacillus clausii fermentation liquor using bacitracin-linked adsorbents, the integrated magnetic separation system exhibited substantially enhanced productivity over expanded bed adsorption when operated at processing velocities greater than 48 m h(-1) . Use of the bacitracin-linked magnetic supports for a single cycle of batch adsorption and subsequent capture by high gradient magnetic separation at a processing rate of 12 m h(-1) resulted in a 2.2-fold higher productivity relative to expanded bed adsorption, while an increase in adsorbent collection rate to 72 m h(-1) raised the productivity to 10.7 times that of expanded bed adsorption . When the number of batch adsorption cycles was then increased to three, significant drops in both magnetic adsorbent consumption (3.6 fold) and filter volume required (1.3 fold) could be achieved at the expense of a reduction in productivity from 10.7 to 4.4 times that of expanded bed adsorption. Bioelectrochemistry, 2002 Jan, 55(1-2), 57 - 9 Affinity interactions on a liposome surface detected by ultrasound velocimetry; Krivanek R et al.; In this work, we performed targeted immobilization of immunoglobulins by means of bacterial S-layer proteins from Bacillus coagulans E38-66/V1 recrystallized on liposomes, which were exploited as immobilization matrix for antibody (Ab)-human IgG . The study of interaction of rabbit or swine anti-human IgG as antigens (Ag) was performed by means of measuring changes of ultrasound velocity . We showed that at a temperature of 25 degrees C, the increment of ultrasound velocity {u} linearly decreased following an increase of concentration of Ag . The decrease of {u} was presumably due to changes of hydration of the membrane due to the binding process . Approximately 10 times lower changes of {u} were observed at 45 degrees C for Ag-Ab interaction as well as for nonspecific interaction of Ag with liposomes covered by S-layer without Ab . No substantial differences in the behaviour of {u} were observed for interactions of human IgG with rabbit or swine anti-human IgG. FEMS Microbiol Lett, 2002 Jan 2, 206(1), 63 - 7 Isolation of a novel, phosphate-activated glutaminase from Bacillus pasteurii; Klein M et al.; In Bacillus pasteurii glutamine is being taken up efficiently by a sodium-dependent uptake system and subsequently hydrolysed to ammonium and glutamate . Concerning the latter process, a catabolic L-glutamine amidohydrolase (glutaminase) was isolated from the cytoplasm of this alkaliphilic bacterium and purified to homogeneity using liquid chromatography . Biochemical and physical parameters of the pure enzyme were examined in detail . Interestingly, analysis of the glutaminase revealed a marked increase in catalytic activity in the presence of phosphate, a property yet restricted to animal glutaminases . This is the first report on the presence of a phosphate-activated glutaminase in bacteria. Mikrobiologiia, 2001 Nov-Dec, 70(6), 820 - 4 {Effect of chitosan oligomer on phage particles and reproduction of phage 1-97A in Bacillus thuringiensis culture}; Kochkina ZM et al.; The causes of bacteriophage 1-97A inactivation by the chitosan oligomer with a polymerization degree of 15 and the influence of the oligomer on the phage reproduction in the culture of Bacillus thuringiensis subsp . galleriae, strain 1-97, were studied . The study of the inactivation kinetics showed that, in 1 h, virtually all chitosan was bound to the phage particles, causing, as evidenced by electron microscopy, DNA release from the phage head, destruction of the phage particles, and agglutination of the phage particles or of their tails in the region of the endplate . High-polymeric chitosan caused more pronounced destruction of the phage particles than the oligomer . It was established that chitosan prevented the production of complete phage particles . One of the mechanisms of such an influence may be the production in the presence of chitosan of phage particles devoid of DNA. Mikrobiologiia, 2001 Nov-Dec, 70(6), 811 - 9 {Characteristics of Bacillus cereus dissociants}; Doroshenko EV et al.; The autoregulation of the phenotypic (populational) variability of the Bacillus cereus strain 504 was studied . The isolated colonial morphotypes of this bacterium were found to differ in their growth characteristics and the synthesis of extracellular proteases . The phenotypic variabilities of vegetative proliferating cells and those germinated from endospores and cystlike refractory cells were different . Bacterial variants also differed in the production of the d1 and d2 factors (the autoinducers of dormancy and autolysis, respectively) and sensitivity to them . The possible role of these factors in the dissociation of microorganisms is discussed. Environ Health Perspect, 2002 Jan, 110(1), 5 - 13 Starlink corn: a risk analysis; Bucchini L et al.; Modern biotechnology has dramatically increased our ability to alter the agronomic traits of plants . Among the novel traits that biotechnology has made available, an important group includes Bacillus thuringiensis-derived insect resistance . This technology has been applied to potatoes, cotton, and corn . Benefits of Bt crops, and biotechnology generally, can be realized only if risks are assessed and managed properly . The case of Starlink corn, a plant modified with a gene that encodes the Bt protein Cry9c, was a severe test of U.S . regulatory agencies . The U.S . Environmental Protection Agency had restricted its use to animal feed due to concern about the potential for allergenicity . However, Starlink corn was later found throughout the human food supply, resulting in food recalls by the Food and Drug Administration and significant disruption of the food supply . Here we examine the regulatory history of Starlink, the assessment framework employed by the U.S . government, assumptions and information gaps, and the key elements of government efforts to manage the product . We explore the impacts on regulations, science, and society and conclude that only significant advances in our understanding of food allergies and improvements in monitoring and enforcement will avoid similar events in the future . Specifically, we need to develop a stronger fundamental basis for predicting allergic sensitization and reactions if novel proteins are to be introduced in this fashion . Mechanisms are needed to assure that worker and community aeroallergen risks are considered . Requirements are needed for the development of valid assays so that enforcement and post market surveillance activities can be conducted. Clin Immunol, 2002 Jan, 102(1), 25 - 7 561del4 defines a novel small deletion hotspot in the interferon-gamma receptor 1 chain; Rosenzweig S et al.; Patients with a dominant small deletion (818del4, hotspot) in the interferon-gamma receptor 1 (IFNGR1) gene (6q23-q24) and increased susceptibility to mycobacterial infections have been recently reported . We describe a female patient homozygous for a 4-bp deletion in exon 5 of IFNGR1 (561del4) who developed postvaccinal disseminated Bacille Calmette-Guerin infection . She was born to unrelated Argentinean parents, each of whom was heterozygous for this mutation . 561del4 has been previously described as a maternally inherited mutation in a compound heterozygous German patient . By single nucleotide polymorphism analysis of the areas surrounding the deletion, we showed the independent inheritance of 561del4 in three heterozygous carriers . Polypurine runs and "direct repeats," previously shown to be associated with areas of recurrent small deletions, were found in the flanking region of 561del4 . The independent inheritance of three identical mutational events defines 561del4 as a new hotspot in the IFNGR1 gene . (c) 2001 Elsevier Science. Zhonghua Kou Qiang Yi Xue Za Zhi, 2000 Sep, 35(5), 365 - 7 {The study of histopathology and bacteriology of coronal pulp tissue in deciduous teeth with deep dentin caries}; Wang X et al.; OBJECTIVE: To study the status of coronal pulp of deciduous teeth with deep caries and causes of pulpal histopathological changes . METHODS: 45 deciduous molars with occlusal deep caries were selected at random . The patients did not receive any antibiotic for one month . The coronal pulpal histopathology was observed with optical microscopic technique . The bacteria in pulp chamber were cultured by the ways of aerobic and anaerobic culture technique . The dentinal tubular structures and bacteria within the tubules of cavity basal part dentin were studied with scanning electron microscopic technique . RESULTS: Coronal pulp tissue of 77.8% of deciduous teeth with deep caries showed chronic inflammation to various degrees or chronic pulpitis with acute attack . There were aerobic and anaerobic mixed infection in pulp chambers of 94.9% deciduous teeth with deep caries . It was found that coccus and bacillus invaded into dentinal tubules of cavity basal part . Coccus located inside tubules with intact tubular walls, while tubular walls were destroyed the bacillus was present . CONCLUSION: There are aerobic and anaerobic mixed infection in pulp chamber of deciduous teeth with deep caries . The coronal pulp tissue shows chronic inflammation to various extent in over 2/3 deciduous teeth with deep caries . Bacillus has stronger destructive power to dental tubules. Biochem Biophys Res Commun, 2002 Jan 11, 290(1), 546 - 51 Compact form of DNA induced by DNA-binding protein HU; Endo T et al.; Interaction of DNA-binding protein HU from Bacillus stearothermophilus (HUBst) with coliphage T2 DNA was investigated by means of a single-duplex DNA chain visualization method using fluorescence microscopy . Fluorescence microscopic images of coliphage T2 DNA molecules were observed as a function of HUBst concentration . The average fluorescence image size of T2 DNA decreased with increase in HUBst concentration to a size comparable to that of a DNA globule induced by polyethylene glycol (PEG) and multivalent cation (MVC) . The change to globule-like DNA proceeded gradually and monotonously, in contrast to the coil-globule transition of DNA induced by PEG and MVC . The histogram of the fluorescence image length was essentially a single-modal one throughout the process of conformational change . These results indicate that the process of shrinking of DNA from a random coil to a globule-like one is not of a transitional nature . The interaction of HUBst with DNA and the mechanism of shrinkage are concluded to be different from those of PEG-induced and MVC-induced coil-globule transition of DNA . (c)2002 Elsevier Science. Extremophiles, 2001 Dec, 5(6), 423 - 9 A catalase-peroxidase from a newly isolated thermoalkaliphilic Bacillus sp . with potential for the treatment of textile bleaching effluents; Gudelj M et al.; A new thermoalkaliphilic bacterium was isolated from a textile wastewater drain and identified as a new Bacillus sp . (Bacillus SF) . Because of its high pH stability and thermostability, a catalase-peroxidase (CP) from this strain has potential for the treatment of textile bleaching effluents . The CP from Bacillus SF was purified to more than 70.3-fold homogeneity using fractionated ammonium sulfate precipitation, hydrophobic interaction, and anion-exchange and gel-filtration chromatography . The native CP had a molecular mass of 165 kDa and was composed of two identical subunits . The isoelectric point of the protein was at pH 6.0 . Peptide mass mapping using matrix-assisted laser desorption ionization-mass spectrometry showed a homology between the CP from Bacillus SF and the CP from Bacillus stearothermophilus . The apparent Km value of the catalase activity for H2O2 was 2.6 mM and the k(cat) value was 11,475 s(-1) . The enzyme showed high catalase activity and an appreciable peroxidase activity with guaiacol and o-dianisidine . The enzyme was stable at high pH, with a half-life of 104 h at pH 10 and 25 degrees C and 14 h at 50 degrees C . The enzyme was inhibited by azide and cyanide, in a competitive manner, but not by the catalase-specific inhibitor 3-amino-1,2,4-triazole. J Immunol, 2002 Jan 15, 168(2), 919 - 25 Influence of Mycobacterium bovis bacillus Calmette-Guérin on antibody and cytokine responses to human neonatal vaccination; Ota MO et al.; The immaturity of the immune system increases the susceptibility of young infants to infectious diseases and prevents the induction of protective immune responses by vaccines . We previously reported that Mycobacterium bovis bacillus Calmette-Guerin (BCG) vaccination induces a potent Th1 response to mycobacterial Ags in newborns . In this study, we evaluated the influence of BCG on the response to unrelated vaccines given in early life . Newborns were randomly allocated to one of three study groups receiving BCG at birth, when infants received their first dose of hepatitis B and oral polio vaccines; at 2 mo of age, when infants received their first dose of diphtheria and tetanus vaccines; or at 4.5 mo of age, when immune responses to vaccines were measured . Administration of BCG at the time of priming markedly increased the cellular and Ab responses to the hepatitis B vaccine, but had only a limited influence on the cytokine response to tetanus toxoid and no effect on the Ab responses to tetanus and diphtheria toxoids . Although BCG induced a potent Th1-type response to mycobacterial Ags, it promoted the production of both Th1- and Th2-type cytokines in response to unrelated vaccines . The effect of BCG was apparent at the systemic level, as it increased the Ab response to oral polio vaccine . These results demonstrate that BCG influences the immune response to unrelated Ags in early life, likely through its influence on the maturation of dendritic cells. J Econ Entomol, 2001 Dec, 94(6), 1589 - 93 Field efficacy and seasonal expression profiles for terminal leaves of single and double Bacillus thuringiensis toxin cotton genotypes; Adamczyk JJ Jr et al.; Examination of commercial Cry1Ac transgenic Bacillus thuringiensis Berliner (Bt) cotton varieties (Bollgard, Monsanto, St . Louis, MO) and an experimental Cry1Ac + Cry2Ab transgenic Bt cotton variety (Bollgard II, Monsanto) for lepidopteran field efficacy was conducted during the 2000 growing season . In addition, a commercially available (Envirologix, Portland, ME) quantification assay (ELISA) was used to measure and profile the expression levels of Cry proteins in two of these varieties {'DP50B, Bollgard'; 'DP50BII, Bollgard II' (Delta & Pine Land, Scott, MS)} . Populations of beet army worms, Spodoptera exigua (Hubner), and soybean loopers, Pseudoplusia includens (Walker), were significantly lower (P < 0.05) in Bollgard II plots compared with Bollgard . Population numbers for fall army worms, Spodoptera frugiperda (J . E . Smith), and salt marsh caterpillars, Estigmene acrea (Drury), were lower in Bollgard II plots compared with Bollgard but means did not differ significantly (P > 0.05) . Single and dual-toxin genotypes remained superior (P < 0.05) compared with conventional cotton against the tobacco budworm, Heliothis virescens (F.) . The addition of Cry2Ab had no significant (P > 0.05) impact on Cry1Ac expression in Bollgard II compared with Cry1Ac expression in Bollgard . Furthermore, throughout the season Cry2Ab was present at much higher levels in the plant compared with Cry1Ac for Bollgard II plants . Possible species-specific reasons for increased efficacy of Bollgard II over Bollgard are discussed. J Econ Entomol, 2001 Dec, 94(6), 1571 - 6 Fitness costs and maternal effects associated with resistance to transgenic cotton in the pink bollworm (Lepidoptera: Gelechiidae); Carriere Y et al.; Transgenic cotton producing a Bacillus thuringiensis (Bt) toxin is widely used for controlling the pink bollworm, Perctinophora gossypiella (Saunders) . We compared performance of pink bollworm strains resistant to Bt cotton with performance of their susceptible counterparts on non-Bt cotton . We found fitness costs that reduced survival on non-Bt cotton by an average of 51.5% in two resistant strains relative to the susceptible strains . The survival cost was recessive in one set of crosses between a resistant strain and the susceptible strain from which it was derived . However, crosses involving an unrelated resistant and susceptible strain indicated that the survival cost could be dominant . Development time on non-Bt cotton did not differ between the two related resistant and susceptible strains . A slight recessive cost affecting development time was suggested by comparison of the unrelated resistant and susceptible strains . Maternal effects transmitted by parents that had eaten Bt-treated artificial diet as larvae had negative effects on embryogenesis, adult fertility, or both, and reduced the ability of neonates to enter cotton bolls . These results provide further evidence that fitness costs associated with the evolution of resistance to Bt cotton are substantial in the pink bollworm. J Econ Entomol, 2001 Dec, 94(6), 1564 - 70 Chronic exposure of the European corn borer (Lepidoptera: Crambidae) to Cry1Ab Bacillus thuringiensis toxin; Chaufaux J et al.; Transgenic corn expressing the insecticidal toxin from Bacillus thuringiensis Berliner is gaining support as an effective control technology for use against lepidopteran pests, particularly European corn borer, Ostrinia nubilalis Hubner (Lepidoptera: Crambidae) . However, there is concern that widespread adoption of transgenic plants will rapidly lead to B . thuringiensis toxin resistance . Thus, long-term selection of O . nubilalis populations with the Cry1Ab B . thuringiensis toxin has been undertaken in several laboratories in the United States and in Europe . We present results from two independent selection experiments performed in laboratories at the University of Nebraska and at the Institut National de la Recherche Agronomique in France . Although the protocols and methods used by the two laboratories were different, the results were comparable . The highest level of resistance occurred at generation 7 (14-fold), generation 9 (13-fold), and generation 9 (32-fold) for three different strains . For each strain, the level of resistance fluctuated from generation to generation, although there were consistently significant decreases in toxin susceptibility across generations for all selected strains . These results suggest that low levels of resistance are common among widely distributed O . nubilalis populations. J Econ Entomol, 2001 Dec, 94(6), 1547 - 52 Different cross-resistance patterns in the diamondback moth (Lepidoptera: Plutellidae) resistant to Bacillus thuringiensis toxin Cry1C; Zhao JZ et al.; Two strains of the diamondback moth, Plutella xylostella (L.), were selected using Cry1C protoxin and transgenic broccoli plants expressing a Cry1C toxin of Bacillus thuringiensis (Bt) . Both strains were resistant to Cry1C but had different cross-resistance patterns . We used 12 Bt protoxins for cross-resistance tests, including Cry1Aa, Cry1Ab, Cry1Ac, Cry1Bb, Cry1C, Cry1D, Cry1E, Cry1F, Cry1J, Cry2Ab, Cry9Aa, and Cry9C . Compared with the unselected sister strain (BCS), the resistance ratio (BR) of one strain (BCS-Cry1C-1) to the Cry1C protoxin was 1,090-fold with high level of cross-resistance to Cry1Aa, Cry1Ab, Cry1Ac, Cry1F, and Cry1J (RR > 390-fold) . The cross-resistance to Cry1A, Cry1F, and Cry1J in this strain was probably related to the Cry1A resistance gene(s) that came from the initial field population and was caused by intensive sprayings of Bt products containing Cry1A protoxins . The neonates of this strain can survive on transgenic broccoli plants expressing either Cry1Ac or Cry1C toxins . The other strain (BCS-Cry1C-2) was highly resistant to Cry1C but not cross-resistant to other Bt protoxins . The neonates of this strain can survive on transgenic broccoli expressing Cry1C toxin but not Cry1Ac toxin . The gene(s) conferring resistance to Cry1C segregates independently from Cry1Ac resistance in these strains . The toxicity of Cry1E and Cry2Ab protoxins was low to all of the three strains . The overall progress of all work has resulted in a unique model system to test the stacked genes strategy for resistance management of Bt transgenic crops. J Econ Entomol, 2001 Dec, 94(6), 1445 - 51 Bollworm (Lepidoptera: Noctuidae) survival on 'Bollgard' and 'Bollgard II' cotton flower bud and flower components; Gore J et al.; Genetically modified cotton, Gossypium hirsutum L., cultivars ('Bollgard') that produce crystalline proteins from Bacillus thuringiensis (Berliner) are valuable tools for managing lepidopteran insect pests in the United States . However, high numbers of bollworm, Helicoverpa zea (Boddie), larvae have been observed feeding in white flowers of these cultivars . Fresh tissue bioassays were conducted to investigate bollworm survival on Bollgard and 'Bollgard II' cottons . Bollworm survival was higher on square and flower anthers than on other floral structures on 'Deltapine 5415' (conventional cotton) and 'NuCOTN 33B' (Bollgard) . Bollworm survival at 72 h was higher on all floral structures from Deltapine 5415 than on corresponding structures from NuCOTN 33B . ELISA tests indicated that CryIA(c) expression varied among plant parts; however, bollworm survival did not correlate with protein expression levels . Trends in bollworm survival on Bollgard II were similar to those on Bollgard and conventional cotton; however, survival was lower on all structures of Bollgard II than on corresponding structures of Bollgard and conventional cotton . These data support field observations of bollworm injury to white flowers and small bolls and provide a better understanding of larval behavior on Bollgard cotton. J Econ Entomol, 2001 Dec, 94(6), 1369 - 77 Dispersal of adult European corn borer (Lepidoptera: Crambidae) within and proximal to irrigated and non-irrigated corn; Hun TE et al.; The European corn borer, Ostrinia nubilalis (Hubner), causes economic damage to corn, Zea mays L., throughout the Corn Belt . Because this insect has become the primary target of Bacillus thuringiensis Berliner (Bt) transgenic corn, current efforts addressing the management of O . nubilalis resistance to Bt corn require information on adult European corn borer dispersal and factors affecting its dispersal . In 1998 we conducted mark-release-recapture, release-recapture, and caged-mating studies to directly measure and compare local dispersal patterns of O . nubilalis adults within and proximal to irrigated and non-irrigated cornfields . Releases of marked adults were made corresponding to the first and second flight of O . nubilalis in eastern Nebraska . Adult dispersal was significantly different between irrigated and non-irrigated cornfields . Released adults tended to remain in and near irrigated cornfields, but dispersed out of and away from non-irrigated cornfields . When released at the edge of the cornfield, neither male nor unmated female O . nubilalis displayed an initial tendency to move out of irrigated corn and into the mixed smooth bromegrass (Bromus inermis Leyss) and broadleaf-weed field edge . Mating efficiency in a late-season cornfield was not significantly different than in dense foxtail (Setaria spp.) . Generally, we found that adult O . nubilalis dispersal may vary depending on variables such as action-site availability and agronomic practices and their interaction with O . nubilalis life history. J Econ Entomol, 2001 Dec, 94(6), 1361 - 8 Interactions between Nosema pyrausta (Microsporidia: Nosematidae) and Bacillus thuringiensis subsp . kurstaki in the European corn borer (Lepidoptera: Pyralidae); Pierce CM et al.; Larval susceptibility to Bacillus thuringiensis was determined for Nosema pyrausta-infected and uninfected European corn borers, Ostrinia nubilalis (Hubner), in bioassays using a commercial formulation of B . thuringiensis subsp . kurstaki, Dipel ES, incorporated into diet . LC50 values for N . pyrausta-infected larvae were significantly lower (P<0.0001) than for uninfected larvae and declined with increasing levels of infection . LC50 values for a 15-d bioassay using field-colony first instars were 0.006 and 0.027 mg of Dipel ES/kg of diet for larvae moderately infected by N . pyrausta and uninfected larvae, respectively . Nosema pyrausta-infected larvae reared on Dipel ES-amended diets produced 70-fold fewer spores (P<0.0001) than larvae reared on standard diet . For example, 15 d after placement as first instars on standard diet, infected field-colony larvae produced 7.6-8.7 million N . pyrausta spores per larva; similar larvae placed on diet containing 0.09 mg of Dipel ES/kg of diet produced 85-103 thousand spores per larva . Infected larvae also weighed less and failed to mature on Dipel ES-amended diets . Increased susceptibility of N . pyrausta-infected larvae to Dipel ES and reduced N . pyrausta spore production in larvae feeding on diet containing Dipel ES suggest that Bt corn will have a direct adverse effect on the survival and continual impact of N, pyrausta as a regulating factor on European corn borer populations. J Econ Entomol, 2001 Dec, 94(6), 1340 - 3 Effects of various insecticides on the development of the egg parasitoid Trichogramma dendrolimi (Hymenoptera: Trichogrammatidae); Takada Y et al.; The toxicity of six insecticides, acephate, methomyl, ethofenprox, cartap, chlorfluazuron, and Bacillus thuringiensis (Bt) was tested on different developmental stages of the egg parasitoid, Trichogramma dendrolimi (Matsumura) . Each of the insecticides tested showed different degrees of toxicity to the parasitoid, Ethofenprox showed the highest toxicity and cartap showed relatively higher toxicity compared with the other insecticides . The development of the parasitoids treated with these two insecticides was normal, similar to that of the control group . Only the emergence of adult wasps from host eggs was disturbed . Emergence of the host, Mamestra brassicae larva was reduced following treatment with ethofenprox, cartap and methomyl . However, adult female wasps, which emerged from host eggs treated with the insecticides had the ability to oviposit normally. Zhonghua Jie He He Hu Xi Za Zhi, 1999 Sep, 22(9), 562 - 3 {Cleaning cavity operation in treating relapse bacillary cavitary pulmonary tuberculosis}; Wei C et al.; OBJECTIVE: To study the effective surgical way for serious relapse multidrug resistant bacillary cavitary pulmonary tuberculosis . METHODS: 104 cases of serious relapse multidrug resistant bacillary cavitary pulmonary tuberculosis were cured by cleaning cavity operation from 1981 to 1998 . The operation contained resecting the back rib covering the cavity, separating the intercostal tissue, cutting the external wall of the cavity, cleaning the cavity thoroughly, scraping the cavity till exposing the fresh tissue, killing the tuberculosis germs with organic acid, filling the cavity with the intercostal muscle, fixing and suturing the muscles . RESULTS: Among the 104 cases, 103 cases (99.0%) were cured, 101 cases (98.1%) were cured after the first operation, one case (1.0%) became better . Among the 98 cases followed up, the rate of returning to work were 96%, no one died or relapsed . CONCLUSIONS: The design of cleaning cavity operation is reasonable and it is the effective method to treat the relapse bacillary cavitary pulmonary tuberculosis, the rate of cure and sputum conversion are promising. Zhonghua Jie He He Hu Xi Za Zhi, 1999 Jun, 22(6), 362 - 3 {Surveillance of primary resistance in 481 cases of pulmonary tuberculosis}; Zhen X et al.; OBJECTIVE: To understand the primary resistance of Mycobacterium tuberculosis in Henan province, and to make scientific prophylactic and treatment strategies . METHODS: Drug resistance surveillance (DRS) was implemented in Henan province according to WHO/IUATLD Guidelines for Surveillance of Drug Resistance in Tuberculosis(1994), and proportional method was used to detect primary resistance in 481 new cases of bacillus-positive pulmonary tuberculosis without history of anti-tuberculosis drug use . RESULTS: The primary drug resistance(PDR) rates were 22.7%, 15.8%, 22.7% and 7.7% for INH, RFP, SM and EMB, respectively . The primary multi-drug resistance (MDR) rates were 2.3%, 5.0%, 0.4% and 5.4% for HR, HRS, HRE and HRSE, respectively . CONCLUSIONS: The results show that the problem of primary resistance of Mycobacterium tuberculosis is serious in Henan province, and uniform management of anti-tuberculosis drugs and implementation of DOTs are needed. Zhonghua Jie He He Hu Xi Za Zhi, 1999 Jul, 22(7), 396 - 8 {Clinical analysis of 90 cases with endobronchial tuberculosis}; Yu W et al.; OBJECTIVE: To explore the clinical features and early definite diagnosis of endobronchial tuberculosis(EBTB) . METHODS: Clinical symptoms, chest X-ray/CT manifestations and fiberoptic bronchoscopic findings of 90 cases with EBTB were analyzed . RESULTS: Main symptoms were as follows: cough in 75 cases, fever in 27, haemoptysis in 24, pleural pain in 18, dyspnea in 11, weight loss in 7, etc . Signs of airway obstruction were rare, localized wheezing was found only in 2 cases . 7 cases in chest X-ray were normal, and only 2 cases in CT were indicative of EBTB . Bronchoscopic results showed exudative lesions in 38% of the cases, granulomatous lesions in 9%, cicatricial lesions in 26%, ulcerative lesions in 17%, and normal-like demonstrations in 11% . Positive results were found in 8 out of 50 by sputum acid-fast staining, 73 out of 86 by bronchoscopic brushing smears, and 17 out of 56 by bronchial biopsies . CONCLUSIONS: The clinical features of EBTB are non-specific, and EBTB can not be excluded only by normal chest X-ray findings . In contrast to CT, bronchoscopy plays an important role in definite diagnosis of the disease . When patients with slight fever of unknown origin or respiratory symptoms do not respond to general treatment, EBTB should be suspected and early fibereoptic bronchoscopy be performed . Fiberoptic bronchoscopic brushing examination for acid-fast bacillus and bronchial biopsy are beneficial to rapid and definite diagnosis of EBTB. J Biomol NMR, 2001 Nov, 21(3), 235 - 48 Joint refinement as a tool for thorough comparison between NMR and X-ray data and structures of HU protein; Raves ML et al.; Joint refinement, i.e., the simultaneous refinement of a structure against both nuclear magnetic resonance (NMR) spectroscopic and X-ray crystallographic data, was performed on the HU protein from Bacillus stearothermophilus (HUBst) . The procedure was aimed at investigating the compatibility of the two data sets and at identifying conflicting information . Wherever important differences were found, such as peptide flips in the main-chain conformation, the data were further analyzed to find the cause . The NMR data showed some errors arising either from the manual interpretation of the spectra or from the incorrect account for spin diffusion . The most important artefact inherent to the X-ray data is the crystal packing of the molecules: the effects range from the limitation of the freedom of the flexible parts of the HUBst molecule to possibly one of the peptide flips. J Egypt Soc Parasitol, 2001 Dec, 31(3), 815 - 22 Study on the effect of Bacillus thuringiensis (B.t . serotype H-14) and its toxins against Cephalopina titillator (Clark), the Egyptian camel myiasis producing dipterous fly; Mazyad SA et al.; Bacillus t . israelensis (B.t . serotype H-14) and its toxins have a marked lethal effect on both the eggs and the newly moulted 3rd stage larvae of Cephalopina titillator . The Bacillus was less effective than Bacillus and its toxins on both the eggs and larvae . On the other hand, the larvae were more affected by both Bacillus and Bacillus and its toxins than the eggs . Prolonged exposure time showed more lethal effect. Anal Chem, 2001 Dec 1, 73(23), 5725 - 31 Tandem mass spectrometry of intact proteins for characterization of biomarkers from Bacillus cereus T spores; Demirev PA et al.; Intact protein biomarkers from Bacillus cereus T spores have been analyzed by high-resolution tandem Fourier transform ion cyclotron resonance mass spectrometry . Two techniques have been applied for excitation of the isolated multiply charged precursor ion species: sustained off-resonance irradiation/collisionally activated dissociation and electron capture dissociation . Fragmentation-derived sequence tags and BLAST sequence similarity proteome database searches allow unequivocal identification of the major biomarker protein with unprecedented specificity . Sequence-specific fragmentation patterns further confirm protein identification . Moreover, methodology combining accurate mass measurements of intact proteins with additional information contained in a proteome database permits tentative assignment of several other protein biomarkers isolated from the B . cereus T spores . We argue that approaches involving tandem MS of protein biomarkers, combined with bioinformatics, can drastically improve the specificity of individual microorganism identification, particularly in complex environments. Appl Environ Microbiol, 2002 Jan, 68(1), 408 - 11 Cloning and characterization of the crystal protein-encoding gene of Bacillus thuringiensis subsp . yunnanensis; Balasubramanian P et al.; Molecular cloning and characterization of a novel cry gene, cry32Aa, of Bacillus thuringiensis subsp . yunnanensis was carried out . The Cry32Aa protein was predicted to have a molecular mass of 139.2 kDa and was found to have an unusual 42-amino-acid-long tail at the C terminus . The cry32Aa gene was localized on the 103-MDa plasmid of the organism . Bioassays showed no toxicity against several moths and mosquitoes . However, it exhibited weak toxicity against larvae of the diamondback moth, Plutella xylostella. Appl Environ Microbiol, 2002 Jan, 68(1), 211 - 8 Cloning, sequence analysis, and expression in Escherichia coli of the gene encoding an alpha-amino acid ester hydrolase from Acetobacter turbidans; Polderman-Tijmes JJ et al.; The alpha-amino acid ester hydrolase from Acetobacter turbidans ATCC 9325 is capable of hydrolyzing and synthesizing beta-lactam antibiotics, such as cephalexin and ampicillin . N-terminal amino acid sequencing of the purified alpha-amino acid ester hydrolase allowed cloning and genetic characterization of the corresponding gene from an A . turbidans genomic library . The gene, designated aehA, encodes a polypeptide with a molecular weight of 72,000 . Comparison of the determined N-terminal sequence and the deduced amino acid sequence indicated the presence of an N-terminal leader sequence of 40 amino acids . The aehA gene was subcloned in the pET9 expression plasmid and expressed in Escherichia coli . The recombinant protein was purified and found to be dimeric with subunits of 70 kDa . A sequence similarity search revealed 26% identity with a glutaryl 7-ACA acylase precursor from Bacillus laterosporus, but no homology was found with other known penicillin or cephalosporin acylases . There was some similarity to serine proteases, including the conservation of the active site motif, GXSYXG . Together with database searches, this suggested that the alpha-amino acid ester hydrolase is a beta-lactam antibiotic acylase that belongs to a class of hydrolases that is different from the Ntn hydrolase superfamily to which the well-characterized penicillin acylase from E . coli belongs . The alpha-amino acid ester hydrolase of A . turbidans represents a subclass of this new class of beta-lactam antibiotic acylases. Appl Environ Microbiol, 2002 Jan, 68(1), 194 - 200 Mutagenic analysis of a conserved region of domain III in the Cry1Ac toxin of Bacillus thuringiensis; Masson L et al.; We used site-directed mutagenesis to probe the function of four alternating arginines located at amino acid positions 525, 527, 529, and 531 in a highly conserved region of domain III in the Cry1Ac toxin of Bacillus thuringiensis . We created 10 mutants: eight single mutants, with each arginine replaced by either glycine (G) or aspartic acid (D), and two double mutants (R525G/R527G and R529G/R531G) . In lawn assays of the 10 mutants with a cultured Choristoneura fumiferana insect cell line (Cf1), replacement of a single arginine by either glycine or aspartic acid at position 525 or 529 decreased toxicity 4- to 12-fold relative to native Cry1Ac toxin, whereas replacement at position 527 or 531 decreased toxicity only 3-fold . The reduction in toxicity seen with double mutants was 8-fold for R525G/R527G and 25-fold for R529G/R531G . Five of the mutants (R525G, R525D, R527G, R529D, and R525G/R527G) were tested in bioassays with Plutella xylostella larvae and ion channel formation in planar lipid bilayers . In the bioassays, R525D, R529D, and R525G/R527G showed reduced toxicity . In planar lipid bilayers, the conductance and the selectivity of the mutants were similar to those of native Cry1Ac . Toxins with alteration at position 527 or 529 tended to remain in their subconducting states rather than the maximally conducting state . Our results suggest that the primary role of this conserved region is to maintain both the structural integrity of the native toxin and the full functionality of the formed membrane pore. Appl Environ Microbiol, 2002 Jan, 68(1), 106 - 13 Role of the single regulator MrsR1 and the two-component system MrsR2/K2 in the regulation of mersacidin production and immunity; Guder A et al.; The lantibiotic mersacidin is an antimicrobial peptide of 20 amino acids which inhibits bacterial cell wall biosynthesis by binding to the precursor molecule lipid II and which is produced by Bacillus sp . strain HIL Y-85,54728 . The structural gene of mersacidin as well as accessory genes is organized in a biosynthetic gene cluster which is located on the chromosome and contains three open reading frames with similarities to regulatory proteins: mrsR2 and mrsK2 encode two proteins with homology to bacterial two-component systems, and mrsR1 shows similarity to a response regulator . Both mrsR2/K2 and mrsR1 were inactivated by insertion of an antibiotic resistance marker . Disruption of mrsR1 resulted in loss of mersacidin production; however, producer self-protection was not impaired . In contrast, inactivation of mrsR2/K2 led to an increased susceptibility to mersacidin whereas biosynthesis of the lantibiotic remained unaffected . Binding of mersacidin to intact cells was significantly enhanced in the mrsR2/K2 knockout mutant . Reverse transcription-PCR analysis from total RNA preparations showed that in contrast to the wild-type strain, the structural genes of the ABC transporter MrsFGE were not transcribed in the knockout mutant . It was therefore concluded that producer self-protection against mersacidin is conferred by the ABC transporter MrsFGE and that the transcription of mrsFGE is regulated by MrsR2/K2, whereas production of the antibacterial peptide is solely activated by MrsR1. Prikl Biokhim Mikrobiol, 2001 Nov-Dec, 37(6), 698 - 701 {Insecticidal activity of Bacillus thuringiensis cells}; Khuzhamshukurov NA et al.; The influence of sixteen different nutrient media on the entomopathogenic activity of three Bacillus thuringiensis strains was studied . The medium composition based on potato, yeast extract, and molasses was optimized . B . thuringiensis No 1 grown on the media No 7 and 9 displayed the highest entomopathogenic activity (94.3 and 90.6%, respectively). Prikl Biokhim Mikrobiol, 2001 Nov-Dec, 37(6), 674 - 7 {Purification and characteristics of alkaline proteinase from alkalophylic Bacillus sp.}; Muderrizade A et al.; Alkaline protease was purified from Bacillus sp . isolated from soil . The pH optimum was 11.5 at 37 degrees C . Calcium divalent cation was effective to stabilize the enzyme especially at higher temperatures . The proteolytic activity was inhibited by active site inhibitors of PMSF (Phenylmethylsulfonyl fluoride), and ions of Mg, Mn, Pb, Li, Zn, Ag, Hg . The enzyme was stable in the presence of some detergents, such as Triton-X-100, Tween-80, SDS (sodium dodecyl sulfate) and EDTA (ethylendiaminetetraacetic acid), pH 11.5 and 37 degrees C for 30 min . The optimum pH was 11.5 at 37 degrees C and the optimum temperature was 62 degrees C at pH 11.5. J Food Prot, 2001 Dec, 64(12), 1935 - 42 Prediction of microbial growth in fresh-cut vegetables treated with acidic electrolyzed water during storage under various temperature conditions; Koseki S et al.; Effects of storage temperature (1, 5, and 10 degrees C) on growth of microbial populations (total aerobic bacteria, coliform bacteria, Bacillus cereus, and psychrotrophic bacteria) on acidic electrolyzed water (AcEW)-treated fresh-cut lettuce and cabbage were determined . A modified Gompertz function was used to describe the kinetics of microbial growth . Growth data were analyzed using regression analysis to generate "best-fit" modified Gompertz equations, which were subsequently used to calculate lag time, exponential growth rate, and generation time . The data indicated that the growth kinetics of each bacterium were dependent on storage temperature, except at 1 degrees C storage . At 1 degrees C storage, no increases were observed in bacterial populations . Treatment of vegetables with AcEW produced a decrease in initial microbial populations . However, subsequent growth rates were higher than on nontreated vegetables . The recovery time required by the reduced microbial population to reach the initial (treated with tap water {TW}) population was also determined in this study, with the recovery time of the microbial population at 10 degrees C being <3 days . The benefits of reducing the initial microbial populations on fresh-cut vegetables were greatly affected by storage temperature . Results from this study could be used to predict microbial quality of fresh-cut lettuce and cabbage throughout their distribution. Epidemiol Mikrobiol Imunol, 2001 Nov, 50(4), 157 - 9 {Human transmission of tuberculosis confirmed by DNA fingerprinting}; Kubin M et al.; By means of fingerprinting based on detection of the insertive sequence IS6110 v DNA of M . tuberculosis predigested by the restrictive enzyme PVUII the authors proved epidemiological associations in three patients with bacillary tuberculosis . The primary source of infection was a 44-year-old man, alcoholic and homeless suffering from tuberculosis diagnosed during an episode of ethylic ebriety, discharged prematurely from hospital because of lack of discipline and drunkenness who died after 8 months with a finding of caseous pneumonia . The first contact person was a 53-year-old women suffering from diabetes and hypertension, an auxiliary worker in the laboratory where she probably was infected during manipulation of the patients infected sputum . The other patients was a 49-year-old man, alcoholic, unemployed where a symptomatic tuberculous finding was detected with general weakness, elevated temperatures and expectoration, who most probably was infected by a patient during accidental contact in his domicile . The profiles of RFLP strains of M . tuberculosis of all three patients had an identical appearance of the DF-1 clone formed by seven copies of IS6110 . This profile was not found in any of the total of 98 strains of M . tuberculosis examined within the framework of the molecular epidemiological prevalence study of patients with bacillary tuberculosis notified in Prague in 1999. Ying Yong Sheng Tai Xue Bao, 2000 Feb, 11(1), 61 - 5 {Species composition and distribution of algae in semi-desert algal crusts}; Hu C et al.; Twenty-two species of algae, including 10 Cyanophyta, 4 Chlorophyta, 6 Bacillariophyta and 2 Euglenophyta, were isolated and identified from the semi-desert algal crusts in Shapotou, Ningxia Autonomous Region . The relationship between algae and soil matrix was studied by grinding and superprobe, and the distribution of algal species in eight layers of crusts was examined under light and scanning electron microscopes . It was found that the algae exhibited a layer-typed distribution in intermicron, and the layers of algal crust from surface to inner were in order of inorganic mineral protection layer(ca . 0.02 mm), alga-enriched layer(0.02-2.5 mm) and sparse algae layer(2.5-5.0 mm) . The filaments of algae were united with clay mineral by the polysaccharides and protein of their sheath, or extended directly into mineral granules to network and band sand and soil particles. Ying Yong Sheng Tai Xue Bao, 2000 Feb, 11(1), 119 - 22 {Effect of Bacillus megaterium on Gluconobacter oxydans in mixed culture}; Feng S et al.; To reveal the relationship between Bacillus megaterium and Gluconobacter oxydans in the mixed culture of vitamin C two-step fermentation, the effect of B . megaterium on the growth of G . oxydans and its synthesizing ability of 2-keto-L-gulonic acid(2KGA) was studied . The bioactive metabolites which could enhance the synthesis of 2KGA were isolated and purified by ultrafiltration, gel chromatography and SDS-polyacrylamide gel electrophoresis . Both the culture supernatant and the cytosol of B . megaterium could promote the proliferation of G . oxydans, and the active component in the culture supernatant was above 100 KDa . The culture suernatant could enhance the conversion of L-sorbose to 2KGA, while the cytosol could not . The active components in B . megaterium culture supernatant had molecular weight of 30-50 KDa and above 100 KDa, and the former was a kind of protein with an apparent molecular weight of about 35 KDa, which consisted of one sort of subunit and contained Fe and Zn elements. J Dent Educ, 2001 Nov, 65(11), 1278 - 80 Disinfection/sterilization of extracted teeth for dental student use; Dominici JT et al.; Extracted human teeth are used in many preclinical courses . While there has been no report of disease transmission with extracted teeth, sterilization of teeth used in the teaching laboratory should be a concern . The purpose of this study was to determine the effectiveness of different sterilization/disinfection methods of extracted human teeth using Bacillus stearothermophilus, a bacteria resistant to heat and frequently used to test sterilizers . In this study, 110 extracted molars with no carious lesions were collected and stored in buffered saline . An endodontic occlusal access preparation was cut into the pulp chamber of each tooth . Pulp tissue in the chamber was removed with a broach . Approximately 1 x 10(5) B . stearothermophilus endospores in culture medium were injected into the pulp chamber, sealed with Cavit G, and then placed in sterile saline for twelve hours . Ten teeth were placed into each of eleven groups . Seven groups were immersed for one week in one of the following solutions: a) sterile saline (control group), b) 5.25% NaOCl, c) 2.6% NaOCl, d) 1% NaOCl, e) 10% buffered formalin, f) 2% gluteraldehyde, g) 0.28% quaternary ammonium . Four additional groups were treated by h) 10% formalin for two days, i) 10% formalin for four days, j) autoclaving at 240 degrees F and 20 psi for twenty minutes, and k) autoclaving at 240 degrees F and twenty psi for forty minutes . Each tooth was then aseptically split and placed in an individual test tube with growth medium . Samples were examined for evidence of growth (turbidity) at forty-eight hours . Only autoclaving for forty minutes at 240 degrees F and 20 psi or soaking in 10 percent formalin for one week were 100 percent effective in preventing growth . A chi-square analysis of the data indicates these two methods were significantly better than all other methods (p<0.001). Mikrobiologiia, 2001 Sep-Oct, 70(5), 636 - 41 {The role of chitinase in antifungal activity of Bacillus sp . 739}; Melen'tiev AI et al.; Investigation of the crude extracellular chitinase of Bacillus sp . 739, an antagonist of phytopathogenic fungi, discerned a relationship between the chitinase and antifungal activities of this bacterium . Purified chitinase lost its ability to inhibit the growth of micromycetes . The antagonistic (antifungal) activity of crude chitinase was found to be located in a low-molecular-weight fraction of the enzyme, which does not possess chitinase activity . Both crude and purified chitinase were able to lyse the cell walls of intact mycelium . Accordingly, it may be inferred that the antagonistic activity of Bacillus sp . 739 against micromycetes is largely determined by low-molecular-weight nonenzymatic substances whereas the role of chitinase is to utilize chitin, which is ubiquitously present in soil. Mikrobiologiia, 2001 Sep-Oct, 70(5), 620 - 8 {Synthesis of anabiosis autoinducers in non-spore-forming bacteria as a mechanism regulating their activity in soil and subsoil sedimentary rocks}; Muliukin AL et al.; Non-spore-forming bacteria of the genera Arthrobacter and Micrococcus, isolated from permafrost subsoil, were found to produce greater amounts of the d1 extracellular factor than closely related collection strains isolated from soil . The effect of this factor, responsible for cell transition to anabiosis, was not species-specific . Thus, the d1 crude preparation isolated from the culture liquid of the permafrost isolate Arthrobacter globiformis 245 produced an effect on the collection strain Arthrobacter globiformis B-1112 and also on Micrococcus luteus and Bacillus cereus . The crude d1 preparation from the permafrost isolate of Arthrobacter differed from the chemical analogue of this factor, 4n-hexylresorcinol, in the level of the induced cell response, which may have resulted from different cell sensitivity to various homologs of alkylhydroxybenzenes contained in the d1 preparation . Thus, additional evidence was obtained indicating that autoregulation of bacterial growth and development is implemented at the level of intercellular interactions in microbial communities . Abundant production of the d1 anabiosis-inducing factors by bacteria isolated from permafrost subsoil is probably a result of special antistress mechanisms responsible for the survival of these bacteria under extreme conditions of natural deep cooling. Mikrobiologiia, 2001 Sep-Oct, 70(5), 602 - 6 {Isolation, purification, and separation of the complex preparation of extracellular proteinases with fibrinolytic and anticoagulant properties from Aspergillus ochraceus 513}; Batomunkueva BP et al.; The extracellular proteinase complex of the microscopic fungus Aspergillus ochraceus 513 was isolated, purified, and separated by affinity chromatography on bacillichin-silochrom and subsequent column chromatography on DEAE-Toyopearl 650 M . The extracellular enzyme of the protein C activator type had a molecular mass of 36.5 kDa and activity close to that of the Agkistrodon snake venom protein C activator . The fibrinolytic and anticoagulant activities of the enzyme were investigated. Antonie Van Leeuwenhoek, 2001 Oct, 80(2), 111 - 8 Production of poly-3-hydroxybutyrate by Bacillus sp . JMa5 cultivated in molasses media; Wu Q et al.; A strain of Bacillus sp . coded JMa5 was isolated from molasses contaminated soil . The strain was able to grow at a temperature as high as 45 degrees C and in 250 g/l molasses although the optimal growth temperature was 35-37 degrees C . Cell density reached 30 g/l 8 h after inoculation in a batch culture with an initial concentration of 210 g/l molasses . Under fed-batch conditions, the cells grew to a dry weight of 70 g/l after 30 h of fermentation . The strain accumulated 25-35%, (w/w) polyhydroxybutyrate (PHB) during fermentation . PHB accumulation was a growth-associated process . Factors that normally promote PHB production include high ratios of carbon to nitrogen, and carbon to phosphorus in growth media . Low dissolved oxygen supply resulted in sporulation, which reduced PHB contents and dry weights of the cells . It seems that sporulation induced by reduced supply of nutrients is the reason that PHB content is generally low in the Bacillus strain. Chemistry, 2001 Nov 5, 7(21), 4651 - 5 Oligosaccharide synthesis by coupled endo-glycosynthases of different specificity: a straightforward preparation of two mixed-linkage hexasaccharide substrates of 1,3/1,4-beta-glucanases; Faijes M et al.; Glycosynthases are engineered glycosidases which are hydrolytically inactive yet efficiently catalyse transglycosylation reactions of glycosyl fluoride donors, and are thus promising tools for the enzymatic synthesis of oligosaccharides . Two endo-glycosynthases, the E134A mutant of 1,3/1,4-beta-glucanase from Bacillus licheniformis and the E197A mutant of cellulase Cel7B from Humicola insolens, were used in coupled reactions for the stepwise synthesis of hexasaccharide substrates of 1,3/1,4-beta-glucanases . Because the two endo-glycosynthases show different specificity, towards laminaribiosyl and cellobiosyl donors, respectively, the target hexasaccharides were prepared by condensation of the corresponding disaccharide building blocks through sequential addition of the glycosynthases in a "one-pot" process . Different strategies were used to achieve the desired transglycosylation between donor and acceptor in each step, and to prevent unwanted elongation of the first condensation product and polymerization (self-condensation) of the donor: 1) selection of disaccharide donors differing in the configuration of the hydroxyl substituent normally acting as acceptor, 2) temporary protection of the polymerizable hydroxyl group of the donor, or 3) addition of an excess of acceptor to decrease the probability that the donor can act as an acceptor . The best procedure involved the condensation of alpha-lactosyl or 4II-O-tetrahydropyranyl-alpha-cellobiosyl fluorides with alpha-laminaribiosyl fluoride, catalyzed by E197A Cel7B, to give tetrasaccharide fluorides, which were then the donors for in situ condensation with methyl beta-cellobioside catalyzed by E134A 1,3/1,4-beta-glucanase . After isolation, the final hexasaccharides Gal/beta4Glcbeta4Glcbeta3Glcbeta4Glcbeta4Glcbeta-OMe and Glcbeta4Glcbeta4Glcbeta3Glcbeta4Glcbeta4-Glcbeta-OMe were obtained in 70-80% overall yields. Korean J Radiol, 2000 Apr-Jun, 1(2), 114 - 7 Disseminated BCG infection in a patient with severe combined immunodeficiency; Han TI et al.; Disseminated mycobacterial infection after bacillus Calmette-Guerin (BCG) vaccination is a very rare disorder, occurring mostly in patients with immunologic deficiency . We report a case of disseminated BCG infection in a 16-month-old girl with severe combined immunodeficiency . Plain radiographs showed multiple osteolytic lesions in the femora, tibiae, humerus, and phalanges . Abdominal sonography and CT scanning revealed multiple nodules in the spleen, and portocaval lymphadenopathy. Acta Crystallogr D Biol Crystallogr, 2002 Jan, 58(Pt 1), 182 - 5 Epub 2001 Dec 21. Expression, purification, crystallization and preliminary crystallographic analysis of a thermostable lipase from Bacillus stearothermophilus P1; Sinchaikul S et al.; The gene encoding a thermostable lipase secreted by Bacillus stearothermophilus P1 has been cloned and overexpressed in Escherichia coli . The recombinant lipase was purified to homogeneity using ammonium sulfate precipitation, anion-exchange chromatography (Poros 20 HQ) and Sephacryl S-200HR . The molecular mass was shown to be 43 209 Da by mass spectrometry . Crystals suitable for X-ray diffraction analysis were obtained by the hanging-drop method of vapour diffusion with ammonium sulfate as the precipitating agent . Determination of the structure by molecular replacement with existing mesophilic lipase structures has proved unrewarding, as there is less than 20% sequence identity with known lipase structures, but preliminary results with heavy-atom soaking indicate that this strategy will allow the structure to be solved . The availability of this new lipase structure will be of particular significance because it will be the first thermostable lipase to be described. FEMS Microbiol Lett, 2001 Dec 18, 205(2), 369 - 74 A protein kinase inhibitor as an antimycobacterial agent; Drews SJ et al.; The protein kinase inhibitor 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H7) was found to inhibit the growth of two different mycobacterial strains, the slow-growing Mycobacterium bovis Bacille Calmette Guerin (BCG) and the fast-growing saprophyte Mycobacterium smegmatis mc2 155, in a dose-dependent manner . While screening for the effect of kinase inhibitors on mycobacterial growth, millimolar concentrations of H7 induced a 40% decrease in the growth of M . bovis BCG when measured as a function of oxidative phosphorylation . This H7-induced decrease in growth was shown to involve a 2-log fold decrease in the viable counts of M . smegmatis within a 48-h period and a 50% reduction in the number of BCG viable counts within a 10-day period . Micromolar concentrations of H7 compound induced a significant decrease in the activity of the Mycobacterium tuberculosis protein serine/threonine kinase (PSTK) PknB . The inhibition of mycobacterial growth as well as the inhibition of a representative M . tuberculosis protein serine/threonine kinase PknB suggests that conventional PSTK inhibitors can be used to study the role that the mycobacterial PSTK family plays in controlling bacterial growth. FEMS Microbiol Lett, 2001 Dec 18, 205(2), 221 - 4 Secretion of a trypsin-like thiol protease by a new keratinolytic strain of Bacillus licheniformis; Rozs M et al.; When cultured in feather-containing broth with a growth optimum of pH 7.0 and 47 degrees C, a Bacillus licheniformis strain exhibited a high chicken feather-degrading activity . A trypsin-like protease was isolated from its ferment broth and was partially characterized . The enzyme was constitutively secreted and was highly active towards N-benzoyl-Phe-Val-Arg-p-nitroanilide as chromogenic substrate . Its pH optimum was 8.5 and it exhibited the highest activity at 52 degrees C . Fractionation on Sephadex G-100 column revealed that its molecular mass was about 42 kDa . The enzyme, which is new for the genus Bacillus, is a thiol protease, as tosyl-L-phenylalanine chloromethyl ketone, tosyl-L-lysine chloromethyl ketone, phenylmethylsulfonyl fluoride and ethylenediamine tetraacetate did not inhibit it, while HgCl2 and para-chloromercuribenzoate lowered its activity. Vet Microbiol, 2002 Feb 4, 84(4), 327 - 36 Vaccination of the brushtail possum (Trichosurus vulpecula) against Mycobacterium bovis infection with bacille Calmette-Guérin: the response to multiple doses; Corner LA et al.; In New Zealand, the brushtail possum (Trichosurus vulpecula) is the principal wildlife vector of bovine tuberculosis . Control of infected possum populations contributes to the control of tuberculosis in domestic livestock . Vaccination is potentially a complementary strategy to population control, but to be cost-effective, administration of the vaccine to possums would need to be from an appropriately designed automatic vaccinator . Possums themselves would activate the vaccinator so that it would deliver an aerosol spray of vaccine . There would be no direct way to prevent possums receiving multiple doses of vaccine . This study examined the effect on protective immunity of repeated vaccination . Captive possums were vaccinated with BCG strain pasteur 1173P2 either 12 times at weekly intervals, twice at 6-weekly intervals, or once . Vaccination was by a combination of intranasal aerosol and conjunctival instillation . Eight weeks after the last dose of vaccine, all possums were challenged intratracheally with Mycobacterium bovis strain 83/6235 . Vaccination induced a significant immune response as measured by the lymphocyte proliferation assay (LPA) . A significant level of protection, as measured by the response to challenge, developed in all the vaccinated possum groups, but protection was greatest in the group vaccinated 12 times . It was concluded that protection would be enhanced if vaccinations were repeated at short intervals (weekly), but no benefit or detriment resulted from revaccination after longer intervals (1-2 months). J Mol Recognit, 2001 Sep-Oct, 14(5), 295 - 9 One-step purification of glucoamylase by affinity precipitation with alginate; Teotia S et al.; It was found that alginate binds to glucoamylase, presumably through the recognition of starch binding domain of the latter . The present work exploits this for purification of glucoamylases from commercial preparation of Aspergillus niger and crude culture filtrate of Bacillus amyloliquefaciens by affinity precipitation technique in a single-step protocol . Glucoamylase is selectively precipitated using alginate as macroaffinity ligand and later eluted with 1.0 M maltose . In the case of A . niger, 81% activity is recovered with 28-fold purification . The purified glucoamylase gave a single band on SDS-PAGE corresponding to 78 kDa molecular weight . The developed affinity precipitation process also works efficiently for purification of Bacillus amyloliquefaciens glucoamylase from its crude culture filtrate, giving 78% recovery with 38-fold purification . The purified preparation showed a major band corresponding to 62 kDa and a faint band about 50 kDa on SDS-PAGE . The latter corresponds to the molecular weight for alpha-amylase of Bacillus amyloliquefaciens . Environ Mol Mutagen, 2001, 38(2-3), 97 - 104 Resistance of spores of Bacillus species to ultraviolet l