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At least 685 records · Page 38Linked to original sources
PubMed1959

Chemical and morphological studies of bacterial spore formation. II. Spore and parasporal protein formation in Bacillus cereus var. alesti.

The development of both the spore and parasporal protein crystal of Bacillus cereus var. alesti was followed using chemical and cytological techniques. The changes which led to the formation of the fore-spore were similar to those already described for Bacillus cereus. However, adjacent to the developing fore-spore a small inclusion became discernible in phase contrast. This protein inclusion during its growth was differentiated from the chromatin and lipid-containing inclusions by sequential staining techniques. During spore and crystal formation no net synthesis of either nucleic acid was detected. Tracer studies with radioactive phosphorus confirmed that the spore chromatin was derived from that in the vegetative cell. These same studies also indicated that a turnover of ribonucleic acid occurred during the sporulation process. During their formation both the spore and crystal incorporated methionine-(35)S from the medium and from cellular material into a bound form. Sequential extractions with alkali and with alkaline-thioglycollate reagent revealed that the solubility characteristics of the mature crystal were possibly related to the presence of intermolecular disulphide bonds which developed after the major synthesis of the crystal was complete. The synthetic nature of sporogenesis and crystal formation is discussed with reference to the concept of "endotrophic" sporulation.

Bacillus cereus↗
PubMed1993

[Fragmentation and solubilization of filamentous appendages of Bacillus cereus spores].

Filamentous appendages were removed from the surfaces of Bacillus cereus spores by treatment with sodium thioglycolate and purified by filtration through a glass microfiber paper and a membrane filter. The basic structure of the isolated appendages resembled that of the native appendages on the spores. The results of electron microscopical analysis, amino acid analysis, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) attested the purity of the preparation. The purified filamentous appendages were fragmented by treatment with 2N sodium hydroxide at 30C but neither with SDS nor mercaptoethanol, and the fragmentation progressed with the prolongation of the treatment period. When observed under an electron microscope, each fragmented appendage appeared to be rod shaped with an apparent axial hole and helical or disk-like arrangement of subunits. In the agar gel immunodiffusion test, a soluble fraction of the appendages obtained by treatment with sodium hydroxide formed only a single precipitation line against the antibody to the intact appendages. The molecular mass of the subunit separated by SDS-PAGE was approximately 10kDa. The 10-kDa protein was distributed over the surface of the intact appendages in immunoelectron microscopy. The antigenicity of the appendage subunits disappeared upon treatment with 2N sodium hydroxide for longer than 24h. Neither intact appendages nor intact spores agglutinated sheep or guinea-pig erythrocytes.

Amino Acids↗
PubMed1996

Feeding induced by pharmacological blockade of fatty acid metabolism is selectively attenuated by hindbrain injections of the galanin receptor antagonist, M40.

Galanin has been shown to stimulate feeding when injected intracranially in rats. Lesion and Fos studies have shown that the neural pathway for feeding stimulated by mercaptoacetate (MA)-induced blockade of fatty acid oxidation includes several structures rich in galanin cell bodies or terminals. In the present experiment, we examined the role of hindbrain galanin in feeding stimulated by MA. We found that galanin (1 nmol) stimulates feeding when injected in the nucleus of the solitary tract (NTS), a site that is crucial for MA-induced feeding, or into the fourth ventricle (4V, 1 or 5 nmol) and that NTS or 4V injections of the galanin receptor antagonist, M40 (1.5 or 5 nmol), completely blocked feeding induced by MA (68 mg/kg). The effect of the M40 appeared to be specific for MA-induced feeding, since M40 did not significantly attenuate either feeding induced by the antimetabolic glucose analog, 2-deoxy-D-glucose (2DG, 100 or 200 mg/kg), or deprivation-induced water intake. Results suggest that feeding induced by decreased fatty acid oxidation relies upon galaninergic terminals in the hindbrain. Furthermore, results indicate that hindbrain neurons involved in MA-induced feeding differ neurochemically from those important for 2DG-induced feeding.

Animals↗
PubMed1984

The enhancement of tumor cell susceptibility to macrophage binding and cytolysis by p-aminobenzoic acid-N-xyloside sodium salt (K-247).

The enhancement of tumor cell susceptibility to macrophage binding and cytolysis by the pretreatment of tumor cells by p-aminobenzoic acid-N-xyloside sodium salt (K-247) was investigated in the C3H/He mouse-syngeneic tumor system. Binding and cytolytic activities of Corynebacterium parvum-activated macrophages were significantly enhanced when target MM-102 and MH-134 cells were pretreated with K-247 at doses of 200 or 400 micrograms/ml, while thioglycollate-elicited macrophages showed much lower binding and lytic activities against K-247 pretreated target cells. No enhancement of these activities were observed when target cells were pretreated with D-xylose, which had no anti-tumor activity. Furthermore, in a binding assay a significant reduction of macrophage binding to target cells by the K-247 pretreated cold competitors was observed. It is suggested that target cell susceptibility to macrophage cytolytic activity might be enhanced by pretreatment with K-247, involving an initially increased target binding.

4-Aminobenzoic Acid↗
PubMedDate not supplied

Rapid determination of Listeria monocytogenes in foods using a resuscitation/selection/kit system detection.

A resuscitation medium consisting of a trypticase soy broth base supplemented with 0.5% yeast extract, 0.25% sodium pyruvate, 0.01% sodium thioglycollate, and 0.1% chicken fat was used in the resuscitation of heat-injured and freeze-injured cells of Listeria monocytogenes. After a resuscitation period of 4-h, the medium was made selective through the addition of nalidixic acid, acriflavin, and cycloheximide. The organisms were incubated in the selectivized medium at 35 degrees C for an additional 16 h. The numbers of resuscitated Listeria monocytogenes cells rose from 10(1) to 10(7) cells/mL in 20 h. Similar numbers of Staphylococcus aureus, Escherichia coli, and Salmonella bonn were grown together with Listeria monocytogenes; these organisms did not inhibit the growth of Listeria monocytogenes nor interfere with its detection by the Listeria-Tek kit system. The resuscitation/selection/kit system (RSK) was compared with the methodology in the Bacteriological Analytical Manual (BAM) for the detection of Listeria monocytogenes in 22 naturally contaminated cheese samples: 8 of these were positive by the BAM system and 12 were positive by the RSK system. The 8 Listeria positives found by the BAM system were positive by the RSK system. All 12 Listeria-presumptive positive samples by the RSK system were confirmed to be Listeria monocytogenes. The use of the RSK system enhanced the recovery of the pathogen, and detection was accomplished within 24 h.

Animals↗
PubMed2006

Sialoadhesin-deficient mice exhibit subtle changes in B- and T-cell populations and reduced immunoglobulin M levels.

Sialoadhesin (Sn, also called Siglec-1 or CD169) is a transmembrane receptor and the prototypic member of the Siglec family of sialic acid binding immunoglobulin-like lectins. It is expressed on specialized subsets of resident macrophages in hematopoietic and lymphoid tissues and on inflammatory macrophages. In order to investigate its function, we generated Sn-deficient mice and confirmed that these mice are true nulls by fluorescence-activated cell sorter analysis and immunohistochemistry. Mice deficient in Sn were viable and fertile and showed no developmental abnormalities. Analysis of cell populations revealed no differences in bone marrow, peritoneal cavity, and thymus, but there was a small increase in CD8 T cells and a decrease in B220-positive cells in spleens and lymph nodes of Sn-deficient mice. Furthermore, in spleen there was a slight decrease in follicular B cells with an increase in numbers of marginal zone B cells. B- and T-cell maturation as well as responses to stimulation with thioglycolate were only slightly affected by Sn deficiency. Immunoglobulin titers in Sn-deficient mice were significantly decreased for immunoglobulin M (IgM) but similar for IgG subclasses. These results suggest a role for sialoadhesin in regulating cells of the immune system rather than in influencing steady-state hematopoiesis.

Animals↗
PubMed1995

In situ accessibility of murine macrophage gangliosides.

Gangliosides are implicated in cell signal transduction. Prior to investigating this phenomenon in macrophages, the in situ accessibility of gangliosides to macromolecules was assessed for peritoneal macrophages isolated from normal C3H/HeN and endotoxin-hyporesponsive C3H/HeJ mice. C3H/HeJ resident and thioglycolate-elicited macrophage ganglioside patterns are the same as normal strains, and no strain differences in galactose oxidase accessibility for resident or thioglycolate-elicited macrophage gangliosides were found. The only gangliosides accessible to galactose oxidase in resident macrophages are GM1a structures. In thioglycolate-elicited macrophages, an additional ganglioside is accessible. For Escherichia coli-activated macrophages, where ganglioside distribution differs between strains, a difference in galactose oxidase-accessible gangliosides also exists. Escherichia coli-activated C3H/HeN patterns show three triplets absent in C3H/HeJ patterns. There were no differences in ganglioside accessibility to Vibrio cholerae sialidase between the thioglycolate-elicited C3H/HeJ and C3H/HeN macrophages. However, despite differences in sialidase-sensitive ganglioside content between E.coli-activated macrophages of these strains, sialidase accessibility for E.coli-activated macrophages was also similar. Sialidase-susceptible GM3 was cryptic in either strain under all conditions examined. The accessibility of murine macrophage gangliosides to galactose oxidase or sialidase was independent of their sialic acid species and chain length of the ceramide fatty acid. With the exception of GM3, major murine macrophage gangliosides are accessible in situ to macromolecules, especially to exogenous pathogenic bacterial sialidase which can alter macrophage cell surface characteristics. Altered macrophage ganglioside accessibility appears sometimes as a consequence, but not a cause, of C3H/HeJ endotoxin hyporesponsiveness.

Animals↗
PubMed2006

Beta-adrenergic-mediated inhibition of feeding by mercaptoacetate in food-deprived rats.

This study investigated the effect of intraperitoneal (IP) injections of the fatty acid oxidation (FAO) inhibitor mercaptoacetate (MA, 45.6 mg/kg) on feeding in food-deprived rats. As previously, MA significantly stimulated feeding in ad libitum-fed rats. MA, however, reduced feeding in 18 and 36 h-fasted rats despite apparently antagonizing the fasting-induced increase in hepatic FAO. To test whether this anorectic effect involves beta-adrenergic stimulation, 36 h-fasted rats were IP injected with the nonspecific beta-adrenergic receptor antagonist propranolol (PROP, 0.5 mg/kg) just before MA injection. PROP attenuated MA's feeding-inhibitory effect, suggesting that MA anorexia is at least partially mediated by beta-adrenergic stimulation. Finally, we evaluated the role of subdiaphragmatic vagal afferent fibers in MA's feeding-inhibitory effect by testing the ability of MA to inhibit food intake in fasted rats after subdiaphragmatic vagal deafferentation (SDA). MA inhibited feeding similarly in SDA rats and sham-operated rats. These data demonstrate that subdiaphragmatic vagal afferents are not necessary for the feeding-inhibitory effect of peripheral MA. These results suggest that the FAO inhibitor MA elicits a feeding-inhibitory effect in fasted rats that is mediated by a different mechanism than its feeding-stimulatory effect.

Adrenergic beta-Antagonists↗
PubMed2004

Absolute quantitation of proteins by a combination of acid hydrolysis and matrix-assisted laser desorption/ionization mass spectrometry.

Quantitation by mass spectrometry is increasingly used to monitor protein levels in biological samples. Most of the current methods are based on the relative comparison of protein quantities but are not suited for the determination of the absolute amount of a given protein. Here we describe a method for the absolute quantitation of proteins that is based on amino acid analysis by matrix-assisted laser desorption/ionization mass spectrometry. Proteins are completely hydrolyzed by acid hydrolysis and then mixed with standards of isotopically labeled amino acids. For the presented study, lysine, leucine, and two different types of labeled arginine were examined as standards. Quantitation of proteins is then achieved by measuring the ratios of labeled and unlabeled amino acids. The method has a sensitivity down to the low-femtomole range and can be applied to quantitate proteins separated by gel electrophoresis. Furthermore, we demonstrate that a mixture of two proteins can be quantitated using two labeled amino acids simultaneously.

Amino Acids↗
PubMed1994

Effect of inhibition of hepatic fatty acid oxidation on metabolic and hormonal responses to exercise in rats.

The purpose of the present study was to assess the effects of 2-mercaptoacetate (MA), an inhibitor of hepatic fatty acid oxidation, on the metabolic and pancreatic hormone response to a prolonged (3 h) swimming exercise. All rats were first adrenodemedullated and were either submitted for 3 weeks to a normal (5% fat) or a medium-fat diet (MF; 18% fat). After being submitted to an exercise swimming habituation programme for one week, rats under both dietary conditions were either injected with a bolus dose of MA (600 mumol/kg; ip) or with a saline solution. MA and saline injected rats were either sacrificed after a 3-h swimming exercise or after a 3-h resting period. Administration of MA was associated with a lower level of beta-hydroxybutyrate after exercise in rats fed the MF diet, higher resting and exercising blood glucose levels in rats fed the MF diet, and higher resting and exercising levels of hepatic glycogen in rats fed a normal diet. There were, however, no significant effects of MA on free fatty acid, insulin, glucagon, epinephrine, and norepinephrine concentrations in both dietary conditions either at rest or after exercise. Therefore, the present data do not provide any evidence that the administration of MA, an inhibitor of hepatic fatty acid oxidation, influences the pancreatic hormonal response to exercise. There was also no evidence of a lowering effect of MA on blood glucose levels during exercise.

3-Hydroxybutyric Acid↗