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At least 145 records · Page 8Linked to original sources
PubMed2006

Spores from mesophilic Bacillus cereus strains germinate better and grow faster in simulated gastro-intestinal conditions than spores from psychrotrophic strains.

The species Bacillus cereus, known for its ability to cause food borne disease, consists of a large variety of strains. An important property for discrimination of strains is their growth temperature range. Psychrotrophic strains can grow well at refrigerator temperatures but grow at 37 degrees C with difficulty. Mesophilic strains on the other hand are unable to grow below 10 degrees C, but grow well at 37 degrees C. Spores of six psychrotrophic and six mesophilic strains were investigated for their ability to survive and grow in simulated gastro-intestinal fluids, mimicking the conditions in the gastro-intestinal tract. The germination potential of psychrotrophic and mesophilic spores in simulated intestinal fluid does not differ much. Under conditions simulating the gastro-intestinal passage, 5 out of 6 mesophilic strains showed growth, and only 2 out of 6 psychrotrophic strains. Temperature (37 degrees C) and simulated gastro-intestinal conditions together influenced germination and growth.

Bacillus cereus↗
PubMed1999

Comparing pollen and spore counts collected with the Rotorod Sampler and Burkard spore trap.

BACKGROUND: The Rotorod Sampler and Burkard spore trap are air-sampling instruments commonly used by allergists in the United States. Although both devices are volumetric, their principles of operation and particle recoveries differ. OBJECTIVE: This review will develop some guidelines for interpreting and comparing pollen counts obtained with these instruments. DATA SOURCES: Investigations examining particle recovery by each device will be reviewed. Five studies where the Rotorod and Burkard were operated in parallel will also be assessed. RESULTS: The Rotorod's theoretical and empirical collection efficiencies are low for particles <10 microm but typically exceed 80% for particles above this threshold. This instrument has traditionally been considered insensitive to wind; experimental data present a mixed picture. The Burkard offers high collection efficiencies, particularly for small particles, when an aerosol's velocity is low. Bi-directional errors in collection efficiency occur as a function of increasing wind speed and particle size. Parallel trapping investigations demonstrated that the Burkard yielded a higher estimate of the atmospheric particle concentration for all particle sizes. Differences were widest for small fungus spores but narrowed for pollen-sized particles. Some recovery differences are readily explained by sampling theory. Other disparities may reflect over-sampling, under-sampling or each device's principles of operation. CONCLUSIONS: Both instruments appear to record the same relative changes in airborne particle concentrations. The Burkard appears to be a superior instrument for sampling particles <10 microm. The Rotorod appears to be equal or superior to the Burkard for collecting particles >10 microm. A rough empirical means for comparing differences in particle recovery is presented.

Air Pollution↗
PubMed2006

Determination of ergosterol in ganoderma spore lipid from the germinating spores of Ganoderma lucidum by high-performance liquid chromatography.

A gradient reversed-phase high-performance liquid chromatography (HPLC) method was developed for the separation and determination of free ergosterol in ganoderma spore lipid (GSL) extracted from the sporoderm-broken germinating spores of Ganoderma lucidum. Sodium hydroxide in methanol was added for the hydrolysis of ergosteryl esters to determine the total content of ergosterol in GSL by HPLC. A 0.04 M concentration of sodium hydroxide in reaction mixtures was appropriate for the complete hydrolysis of ergosteryl esters without a significant loss of ergosterol during saponification. In addition, the ergosterol content in four commercial GSL softgel supplements from four different firms was determined. The results showed that the ergosterol content in these samples had significant differences. Ergosterol content may be a suitable marker for evaluating the quality of GSL products.

Chromatography, High Pressure Liquid↗
PubMed1985

Purification and properties of spore-lytic enzymes from Clostridium perfringens type A spores.

Spores of Clostridium perfringens contain at least two spore-lytic enzymes active in hydrolysing cortical peptidoglycan. One enzyme has been purified 1800-fold and has a molecular weight of 17 400 determined from chromatography on Sephadex G-75. Two protein bands were apparent after SDS-PAGE. The isolated enzyme was investigated for response to temperature, pH, ionic strength and enzyme inhibitors, and for mode of action. A second enzyme activity, differing from the first in apparent molecular weight (29 800) as determined by gel exclusion chromatography, and also in its pH optimum and activity on cortical substrate, was also isolated, although not purified to the same extent.

Chromatography, Gel↗
PubMed1977

Comparative dose-survival curves of representative Clostridium botulinum type F spores with type A and B spores.

Radiation survival data of proteolytic (Walls 8G-F) and non-proteolytic (Eklund 83F) type F spores of Clostridium botulinum were compared with dose-response data of radiation-resistant type A (33A) and B (40B) spores. Strain Eklund 83F was as resistant as strain 33A, whereas strain Walls 8G-F was the most sensitive of the four strains tested. The methods suggested for computing both an initial shoulder and a D value for the dose-survival curves yielded results comparable to the graphic techniques used to obtain these two parameters.

Cesium Radioisotopes↗
PubMed1994

Molecular characterization of Frankia microsymbionts from spore-positive and spore-negative nodules in a natural alder stand.

Among the Frankia strains capable of establishing a nitrogen-fixing symbiosis with actinorrhizal plants, in planta sporangial formation is not universal and has led to the distinction between spore-positive (Sp+) and spore-negative (Sp-) nodules. Numerous Frankia strains have been isolated in pure culture from Sp+ nodules of different host plants, but, although they were able to reinfect their respective host plant, none of them was able to differentiate endophytic sporangia under laboratory conditions. The first step of this study was to demonstrate, at the molecular level, the existence of specific Sp+ strain genotypes differing from Sp- strain genotypes in a single alder stand. In a second step, by way of PCR amplification and sequencing of the PCR products, we have characterized oligonucleotide primers specific for the genus Frankia and for each of the two types of Frankia microsymbionts able, or not, to differentiate sporangia inside natural green alder nodules. These primers applied in PCRs with DNA extracted from nodules confirmed the morphological identification and revealed the presence of nodules colonized by both types of actinomycetes. Finally, a preliminary PCR study was conducted with DNA extracted directly from soil samples which permitted checking the rhizosphere of Sp+ and Sp- nodules for the presence of the corresponding strains.

Actinomycetales↗
PubMed1995

Immunoelectron microscopic localization of one of the spore germination proteins, GerAB, in Bacillus subtilis spores.

Ultrastructural localization of GerAB, one of the proteins of Bacillus subtilis spores related to L-alanine-initiated germination, was investigated by immunoelectron microscopy with antipeptide (residues 61 to 80 of GerAB) antiserum and a colloidal gold-immunoglobulin G complex. Immunogold particles were visualized in the boundary region between the cortex and coat of dormant spores, and they were broadly dispersed into the cortex region after germination.

Antibodies, Bacterial↗
PubMed1996

A germination-specific spore cortex-lytic enzyme from Bacillus cereus spores: cloning and sequencing of the gene and molecular characterization of the enzyme.

A gene (sleB) encoding a 24-kDa germination-specific spore cortex-lytic enzyme, probably an N-acetylmuramyl-L-alanine amidase, was cloned from Bacillus cereus, and its nucleotide sequence was determined. It was indicated that the enzyme is produced as a 259-residue protein with a signal sequence of 32 residues and is present in dormant spores in its active form. Sulfhydryl reagents inactivated the enzyme, but mutation of a single cysteine of the protein, Cys-258, to Gly did not cause complete inactivation of the enzyme, suggesting that the residue does not function as the catalytic center of enzyme.

Amidohydrolases↗
PubMed1978

Quantitative recovery of spores from thermophilic spore papers.

A homogeniser (Stomacher 400) which does not damage bacteria has been used to produce an homogenate of spore papers, from which it was possible to make quantitative assessments of viable spores recoverable from such papers before and after exposure to sterilising procedures.

Agar↗
PubMed2005

[Comparison of the immunomodulatory effects of spore polysaccharides and broken spore polysaccharides isolated from Ganoderma lucidum on murine splenic lymphocytes and peritoneal macrophages in vitro].

OBJECTIVE: To compare the immunomodulatory effects of spore polysaccharides (Gl-SP) and broken spore polysaccharides (Gl-BSP) isolated from Ganoderma lucidum(Leyss et Fr.) Karst. on murine splenic lymphocytes and peritoneal macrophages in vitro. METHODS: Mixed lymphocyte culture reaction (MLR), lymphocyte proliferation in the presence or absence of mitogen, and the cytotoxic activity of splenic natural killer (NK) cells were detected with MTT assay in vitro. The percentage of phagocytosis of neutral red (NR) by mouse peritoneal macrophages was detected by colorimetric assay. Splenic T-lymphocyte subpopulations were measured with flow cytometry(FCM). IL-2, IFN-gamma and TNF-alpha in the culture supernatants were detected by ELISA and biological assay. Nitric oxide (NO) production was examined by Griess reaction. RESULTS: At the concentration range of 0.2-12.8 mg/L, Gl-SP and Gl-BSP were shown to increase lymphocyte proliferation in the presence or absence of mitogen, enhance NK cytotoxic activity, augment the production of TNF-alpha and NO in Gl-SP- or Gl-BSP-activated macrophages, as well the percentage of phagocytosis of NR by macrophages in vitro. Both Gl-SP and Gl-BSP could promote MLR, however, at the dose of 12.8 mg/L, Gl-BSP showed higher activity than Gl-SP in the proliferation of lymphocytes. These two kinds of polysaccharide could significantly increase the secretion of IL-2 and IFN-gamma in doublejway MLR at the concentrations of 0.2-12.8 mg/L, but Gl-BSP had stronger effects than Gl-SP at the same concentrations. Both Gl-SP and Gl-BSP could increase the ratio of T-lymphocyte subpopulations in double-way MLR. At the concentrations of 0.2-12.8 mg/L or 3.2-12.8 mg/L, Gl-BSP demonstrated more significant activity in increasing the percentage of the CD4(+) or CD8(+) subset than Gl-SP. At the concentrations of 0.2-0.8 mg/L, the ratio of the CD4(+) and CD8(+) subset in the Gl-BSP treated group was higher than that of the Gl-SP treated group. CONCLUSION: Gl-SP and Gl-BSP have similar immunomodulatory effects in vitro, as though the immunomodulatory effects of Gl-BSP are stronger than that of Gl-SP.

Adjuvants, Immunologic↗
PubMed2000

The effect of windspeed on pollen and spore counts collected with the Rotorod Sampler and Burkard spore trap.

BACKGROUND: A recent review article presented conflicting evidence concerning the relationship between the Rotorod Sampler's particle collection efficiency and windspeed. Conventional wisdom holds that its collection efficiency is not affected by windspeed; experimental data paint a mixed picture. OBJECTIVES: This brief addendum will present additional data concerning the relationship between the Rotorod's collection efficiency and windspeed. This information will be contrasted with similar data for the Burkard spore trap. DATA SOURCE: Data obtained from an obscure technical report published by Ogden et al. in 1969 will be assessed. RESULTS: The Rotorod's collection efficiency appeared to be greatest at moderate windspeeds; lower efficiencies occurred at both lower and higher windspeeds. Maximum changes in collection efficiency ranged from 29% to 39% over the six windspeeds employed. The Burkard exhibited the opposite relationship between collection efficiency and windspeed. Collection efficiency was lowest at moderate windspeeds and higher at both lower and higher windspeeds. Maximum changes in collection efficiency ranged from 38% to 142%. CONCLUSIONS: Four principal conclusions emerged. First, these newly rediscovered data demonstrate that the Rotorod's collection efficiency is affected by windspeed. These results appear to overturn the conventional wisdom which holds that this device is insensitive to wind. Second, the Rotorod is less affected by windspeed than the Burkard. The maximum change in collection efficiency uncovered for the former device was 39%; this represented the minimum maximum change for the latter device. Third, the Rotorod and Burkard are affected by windspeed in fundamentally different ways. The former device's collection efficiency is greatest at moderate windspeeds and lower at both lower and higher windspeeds. The opposite effect occurs with the latter device. Last, the previous two findings may partially explain some of the differences in particle recovery by the two devices noted in the parent publication. Figures and equations presented in this addendum indicate which circumstances are likely to produce these differences.

Air Pollution↗
PubMed2000

The N-terminal prepeptide is required for the production of spore cortex-lytic enzyme from its inactive precursor during germination of Clostridium perfringens S40 spores.

A spore cortex-lytic enzyme of Clostridium perfringens S40 is synthesized during sporulation as a precursor consisting of four domains. After cleavage of an N-terminal preregion and a C-terminal proregion, inactive proenzyme (termed C35) is converted to active enzyme by processing of an N-terminal prosequence with germination-specific protease (GSP) during germination. The present results demonstrated that the cleaved N-terminal prepeptide remained associated with C35. After the isolated complex was denatured and dissociated in 6 M urea solution, removal of urea regenerated a prepeptide-C35 complex which produces active enzyme when incubated with GSP. However, isolated C35 alone could not be activated by GSP. The prepeptide-C35 complex was more heat stable than active enzyme. Thus, non-covalent attachment of the prepeptide to C35 is required to assist correct folding of C35 and to stabilize its conformation, suggesting that the prepeptide functions as an intramolecular chaperone. Recombinant proteins, which have prepeptide covalently bonded to C35, were processed by GSP as well as the in vivo prepeptide-C35 complex, and the full length of the N-terminal presequence was needed to fulfil its role. Although the C-terminal prosequence is present as an independent domain which is not involved in the activation process of the enzyme, it appears that the N-terminal prosequence contributes to the regulation of enzyme activity as an inhibitor of the enzyme.

Amino Acid Sequence↗
PubMed1980

Studies on the bacterial spore coat. (7) Properties of alkali-soluble components from spore coat of Bacillus megaterium.

Properties of alkali-soluble components from the spore coat of Bacillus megaterium were examined by physicochemical methods. They were composed of acidic polypeptides of various molecular weights with small amounts of phosphorus and sugar. They were allowed to dissociate to unit components by incubation with SDS. The major component was partially purified by gel filtration, and shown to have a mean molecular weight of about 11,000.

Bacillus megaterium↗
PubMed1998

Structure and mechanism of action of the protease that degrades small, acid-soluble spore proteins during germination of spores of Bacillus species.

The germination protease (GPR) of Bacillus megaterium initiates the degradation of small, acid-soluble proteins during spore germination. Trypsin treatment of the 46-kDa GPR zymogen (termed P46) removes an approximately 15-kDa C-terminal domain generating a 30-kDa species (P30) which is stable against further digestion. While P30 is not active, it does autoprocess to a smaller form by cleavage of the same bond cleaved in conversion of P46 to the active 41-kDa form of GPR (P41). Trypsin treatment of P41 cleaves the same bond in the C-terminal part of the protein as is cleaved in the P46-->P30 conversion. While the approximately 29-kDa species generated by trypsin treatment of P41 is active, it is rapidly degraded further by trypsin to small inactive fragments. These results, as well as a thermal melting temperature for P41 which is 13 degreesC lower than that for P46 and the unfolding of P41 at significantly lower concentrations of guanidine hydrochloride than for P46, are further evidence for a difference in tertiary structure between P46 and P41, with P46 presumably having a more compact stable structure. However, circular dichroism spectroscopy revealed no significant difference in the secondary structure content of P46 and P41. The removal of approximately 30% of P46 or P41 without significant loss in enzyme activity localized GPR's catalytic residues to the N-terminal two-thirds of the molecule. This finding, as well as comparison of the amino acid sequences of GPR from three different species, analysis of several site-directed GPR mutants, determination of the metal ion content of purified GPR, and lack of inhibition of P41 by a number of protease inhibitors, suggests that GPR is not a member of a previously described class of protease.

Amino Acid Sequence↗
PubMed2002

Direct H atom abstraction from spore photoproduct C-6 initiates DNA repair in the reaction catalyzed by spore photoproduct lyase: evidence for a reversibly generated adenosyl radical intermediate.

Spore photoproduct (SP) lyase, which catalyzes the direct reversal of SP (5-thyminyl-5,6-dihydrothymine) to thymine monomers, is the only identified nonphotoactivatable pyrimidine dimer lyase. Unlike DNA photolyase, SP lyase does not contain a flavin cofactor and does not require light for activation. Instead, preliminary studies point to the presence of an iron-sulfur cluster in SP lyase and the requirement for S-adenosylmethionine (AdoMet) for catalytic activity, suggesting that SP lyase belongs to the growing group of iron-sulfur cluster and AdoMet-dependent radical enzymes. Here we provide evidence for the role of AdoMet as a reversible deoxyadenosyl radical generator, which initiates repair by hydrogen atom abstraction from C-6 of SP. Reaction of 6-(3)H-SP, but not methyl-(3)H-SP, with SP lyase and AdoMet results in transfer of (3)H to AdoMet, while no tritiated 5'-deoxyadenosine is observed. When 5'-tritiated AdoMet is used in the reaction with unlabeled SP, transfer of (3)H into the repaired thymine monomers is observed. These results point to the reversible generation of a 5'-deoxyadenosyl radical intermediate, which reacts directly with the DNA lesion to initiate a radical-mediated beta-scission. We also demonstrate that AdoMet is a catalytic cofactor that is not consumed during turnover. Together, these results support a novel radical-based mechanism for the repair of UV-induced DNA damage.

DNA Repair↗
PubMed1994

Phylogenetic interrelationships of round-spore-forming bacilli containing cell walls based on lysine and the non-spore-forming genera Caryophanon, Exiguobacterium, Kurthia, and Planococcus.

The 16S rRNA gene sequences of "Bacillus aminovorans" and several species considered to be phylogenetically related to the group 2 bacilli of Ash et al. (C. Ash, J. A. E. Farrow, S. Wallbanks, and M. D. Collins, Lett. Appl. Microbiol. 13:202-206, 1991) were determined. A comparative analysis of the sequence data revealed that the round-spore-forming group 2 bacilli, together with some asporogenous taxa (the genera Caryophanon, Exiguobacterium, Kurthia, Planococcus), form a phylogenetically distinct cluster that is only remotely related to Bacillus subtilis, the type species of the genus Bacillus. Within this cluster, planococci, kurthiae, Caryophanon spp., and two lines defined by Bacillus sphaericus and Bacillus fusiformis and by Sporosarcina ureae, Bacillus pasteurii, Bacillus globisporus, and Bacillus psychrophilus were found to be distinct genera. Exiguobacterium aurantiacum and Brevibacterium acetylicum were found to form a distinct clade, which was peripherally related to this cluster. "B. aminovorans" exhibited no specific relationship with the group 2 bacilli or with any of the other reference species examined.

Bacillus↗