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Biomedical subjects

A A Beharka

Publications and source records attributed to A A Beharka.

At least 19 recordsLinked to original sources
PubMed1998

Salmonella infections in the absence of the major histocompatibility complex II.

We examined the pathogenesis of the facultative intracellular bacterium, Salmonella typhimurium in MHCII-/-, C2D knock-out mice, and wild-type C57BL/6J mice. The MHCII knock-out shortened the kinetics of animal death and reduced the dose of S. typhimurium needed to kill mice. We measured the physiological and cytokine responses of both mouse strains after S. typhimurium injection. Animal weight loss, spleen weights, liver weights, thymus weights, and serum corticosterone concentrations were comparable after injection with several doses of bacteria. The only physiological differences observed between the two strains were observed 3 days after injection of the highest dose of bacteria tested. Serum concentrations of tumor necrosis factor alpha, interleukin-2, and interleukin-6 increased in a dose-dependent fashion irrespective of mouse MHCII expression. Therefore, even in the absence of MHCII, mice are able to mount relatively normal physiological and immunological responses. Consistent with these normal responses, an increased percentage of MHCII-/- mice, primed with a low dose of bacteria 13 days earlier, were able to survive a lethal challenge of Salmonella compared with unprimed controls. Lastly, C2D mice had significantly higher serum interleukin-10 concentrations than C57BL/6J mice 48 h after infection with all doses of S. typhimurium. C2D macrophages also secreted significantly more IL-10 and less NO and O2- after lipopolysaccharide or phorbol ester stimulation in vitro than wild-type macrophages.

Animals↗
PubMed1998

Macrophage cell lines derived from major histocompatibility complex II-negative mice.

Two bone-marrow-derived macrophage cell lines, C2D and C2Dt, were isolated from major histocompatibility class II negative knock-out mice. The C2D cell line was stabilized by continuous culture in colony-stimulating factor-1 and the C2Dt cell line was transformed with SV40 virus large T antigen. These cells exhibited phenotypic properties of macrophages including morphology and expression of Mac 1 and Mac 2 cell surface molecules. These cells also had comparable growth to the bone-marrow-derived macrophage cell line B6MP102. These new cell lines were not spontaneously cytotoxic and were only capable of modest killing of F5b tumor cells when stimulated with LPS and interferon-gamma, but not when stimulated with LPS alone or with staphylococcal exotoxin. C2D and C2Dt cells phagocytosed labeled Staphylococcus aureus similarly to B6MP102 cells but less well than C2D peritoneal macrophages. These cell lines secreted interleukin-6, but not tumor necrosis factor or nitric oxide in response to LPS or staphlococcal enterotoxins A or B C2D(t) cells were tumorigenic in C2D and C57BL/6J mice but C2D cells were not. These data suggest that macrophage cell lines can be established from bone marrow cells of major histocompatibility complex II-negative mice.

Animals↗
PubMed1998

Beta-carotene-induced enhancement of natural killer cell activity in elderly men: an investigation of the role of cytokines.

We showed previously that natural killer (NK) cell activity is significantly greater in elderly men supplemented with beta-carotene than in those taking placebo. In an attempt to determine the mechanism of beta-carotene's effect, we analyzed the production of NK cell-enhancing cytokines (interferon alpha, interferon gamma, and interleukin 12). Boston-area participants in the Physicians' Health Study (men aged 65-88 y; mean age, 73 y) who had been supplemented with beta-carotene (50 mg on alternate days) for an average of 12 y were enrolled in a randomized, placebo-controlled, double-blind study. Elderly subjects taking beta-carotene supplements had significantly greater plasma beta-carotene concentrations than those taking placebo. Beta-carotene-supplemented elderly men had significantly greater NK cell activity than did elderly men receiving placebo. Percentages of NK cells (CD16+CD56+) were not significantly different between the beta-carotene and placebo groups. Production of interleukin 12, interferon alpha, or concanavalin A-stimulated interferon gamma by cultured peripheral blood mononuclear cells was not significantly different between beta-carotene-supplemented elderly and those taking placebo. Our results indicate that beta-carotene-induced enhancement of NK cell activity is not mediated by changes in percentages of CD16+CD56+ NK cells nor through up-regulation of interleukin 12 or interferon alpha.

Aged↗
PubMed1998

Age-associated increase in PGE2 synthesis and COX activity in murine macrophages is reversed by vitamin E.

We previously showed that increased macrophage and PGE2 production with age is due to enhanced cyclooxygenase (COX) activity and COX-2 expression. This study determined the effect of vitamin E supplementation on macrophage PGE2 synthesis in young and old mice and its underlying mechanism. Mice were fed 30 or 500 parts per million vitamin E for 30 days. Lipopolysaccharide (LPS)-stimulated macrophages from old mice produced significantly more PGE2 than those from young mice. Vitamin E supplementation reversed the increased PGE2 production in old mice but had no effect on macrophage PGE2 production in young mice. In both LPS-stimulated and unstimulated macrophages, COX activity was significantly higher in old than in young mice at all intervals. Vitamin E supplementation completely reversed the increased COX activity in old mice to levels comparable to those of young mice but had no effect on macrophage COX activity of young mice or on COX-1 and COX-2 protein or COX-2 mRNA expression in young or old mice. Thus vitamin E reverses the age-associated increase in macrophage PGE2 production and COX activity. Vitamin E exerts its effect posttranslationally, by inhibiting COX activity.

Aging↗
PubMed1998

Effect of Aspergillus oryzae extract alone or in combination with antimicrobial compounds on ruminal bacteria.

The effect of an Aspergillus oryzae fermentation extract on the growth rates of pure cultures of ruminal bacteria was determined. Bacteria were grown in an anaerobic ruminal fluid and carbohydrate medium. A sterile filtrate made with 10% A. oryzae was added to the medium at 2 or 5% (vol/vol) to provide a final A. oryzae concentration of 2 or 5 mg/ml, respectively. The filtrate had no effect on the growth rates of 10 of the 19 ruminal bacteria tested; however, the filtrate increased the growth rates of the bacteria that digest fiber, Ruminococcus albus and Fibrobacter succinogenes, and the bacteria that utilize lactate, Megasphaera elsdenii, Selenomonas lactilytica, and Selenomonas ruminantium. No differences in growth rate were detected between the two concentrations of A. oryzae filtrate. We also investigated the interactions between A. oryzae and antimicrobial compounds on the growth rates of six species of ruminal bacteria that had shown positive responses or no response to the filtrate. The addition of A. oryzae filtrate to medium containing chlortetracycline or neomycin tended to diminish the negative effects of those compounds on the growth rates of some ruminal bacteria, although the bacteria had no positive growth response to the filtrate alone. In contrast, the combination of A. oryzae filtrate and tylosin decreased the growth rate of Sel. ruminantium D. These results indicated that A. oryzae stimulates growth of some bacteria that digest fiber and ferment lactate in the rumen and interacts positively or negatively with certain antimicrobial feed additives.

Animal Feed↗
PubMed1998

Effects of form of the diet on anatomical, microbial, and fermentative development of the rumen of neonatal calves.

Eight neonatal, Holstein bull calves were paired by birth date and birth weight and randomly assigned to either a finely ground or unground control diet (chopped hay and rolled grain) to study the effects of the physical form of the diet on anatomical, microbial, and fermentative development of the rumen. The diets varied in particle size but were identical in composition (25% alfalfa hay and 75% grain mix). Calves were fed milk at 8% of birth weight daily until weaning. Feed intake was equalized for each pair of calves. Ruminal fluid samples were collected from ruminal cannulas to determine pH, fermentation products, and buffering capacity and to enumerate bacteria. Calves were slaughtered at 10 wk of age, and weights of the full and empty reticulorumen, abomasum, and omasum were recorded. Ruminal tissue samples were taken to assess papillary development by morphometric measurements. Calves had similar body weights at wk 10. Ruminal pH was affected by age and was lower for calves fed the ground diet. Total anaerobic bacterial counts were not affected by the physical form of the diet; however, calves fed the ground diet had lower numbers of cellulolytic bacteria and higher numbers of amylolytic bacteria than did calves fed the unground diet. Physical form of the diet did not affect the weights of the reticulorumen whether full or empty. However, calves fed the ground diet had heavier omasum weights, both full and empty. Physical form of the diet affected papillary size and shape but did not influence the muscle thickness of rumen. Results indicated that the physical form of the diet had a significant influence on the anatomical and microbial development of the forestomac and, therefore, might influence future performance.

Animal Feed↗
PubMed1997

Enhanced expression of inducible cyclooxygenase with age in murine macrophages.

Macrophages (Mphi) from old mice produce more PGE2 than those from young mice, contributing to the dysregulation of the immune and inflammatory responses with age. This study was conducted to determine the mechanisms of the age-associated increase in Mphi PGE2 production. PGE2 production is influenced by the availability of the substrate arachidonic acid and by activity of the enzyme cyclooxygenase (Cox). We demonstrate that when the substrate is not the limiting factor, Mphi from old mice have significantly higher LPS-stimulated Cox activity than young mice, indicating that the age-associated increase in PGE2 production is due to increased enzyme activity and not to changes in substrate level. Cox activity is determined by the enzyme level and requires hydroperoxide for activation. Of the two Cox isoforms, Cox 1 is constitutively expressed in nearly all cells; whereas Cox 2 is induced by a wide range of ligands. Analysis of accumulated and de novo synthesis of constitutive Cox 1 and inducible Cox 2 proteins showed no age-related difference in Cox 1 protein levels, but Mphi from old mice had higher accumulated and newly synthesized LPS-stimulated Cox 2 protein levels than young mice. Furthermore, Mphi from old mice had higher LPS-stimulated levels of Cox 2 mRNA compared with those from young mice. Clearly, the age-associated increase in LPS-stimulated PGE2 production is due to increased Cox activity resulting from higher Cox 2 protein and mRNA expression. These findings have significant implications for age-associated immune and inflammatory dysregulation as well as the development of preventive and therapeutic strategies against them.

Aging↗
PubMed1997

Macrophage prostaglandin production contributes to the age-associated decrease in T cell function which is reversed by the dietary antioxidant vitamin E.

The aging process is associated with a decline in T cell-mediated immunity, including decreased interleukin (IL)-2 production and mitogen-induced T cell proliferation. Because macrophages (M phi) from old mice have higher production of prostaglandin (PG) E2 than young mice, and PGE2 has been shown to suppress T cell-mediated function, we hypothesized that increased production of PGE2 would contribute to decreased T cell function with aging and that decrease in PGE2 production by dietary antioxidants would enhance T cell-mediated function. Experiments were conducted in which combinations of purified M phi and T cells (> 95% pure) from young or old C57BL/6N1A mice were cultured together. Co-cultures containing T cells and M phi from old mice had reduced ConA-stimulated proliferation and IL-2 secretion than those consisting of T cells and M phi from young mice. Addition of M phi from old mice suppressed proliferation and IL-2 secretion by T cells from young mice. Likewise, T cells from old mice secreted more IL-2 when cultured with M phi from young mice compared to those cultured with M phi from old mice. Addition of PGE2, at concentrations produced by old M phi, decreased proliferation and IL-2 production by young but not old T cells. Neither addition of H2O2 at physiological levels, nor catalase changed the response of cultures from young or old mice. However, addition of indomethacin and the antioxidant nutrient vitamin E, both of which decreased PGE2 production, improved T cell proliferation and IL-2 production. These experiments demonstrate that increased production of PGE2 by M phi contributes to the age-associated decline in T cell function. Vitamin E improves T cell responsiveness in old mice mostly by reducing M phi PGE2 production, although a direct effect of vitamin E on T cells was also observed.

Aging↗
PubMed1995

Staphylococcal enterotoxins bind H-2Db molecules on macrophages.

We screened a panel of monoclonal antibodies against selected macrophage cell surface molecules for their ability to inhibit enterotoxin binding to major histocompatibility complex class II-negative C2D (H-2b) macrophages. Two monoclonal antibodies, HB36 and TIB126, that are specific for the alpha 2 domain of major histocompatibility complex class I, blocked staphylococcal enterotoxins A and B (SEA and SEB, respectively) binding to C2D macrophages in a specific and concentration-dependent manner. Inhibitory activities were haplotype-specific in that SEA and SEB binding to H-2k or H-2d macrophages was not inhibited by either monoclonal antibody. HB36, but not TIB126, inhibited enterotoxin-induced secretion of cytokines by H-2b macrophages. Lastly, passive protection of D-galactosamine-sensitized C2D mice by injection with HB36 antibody prevented SEB-induced death. Therefore, SEA and SEB binding to the alpha 2 domain of the H-2Db molecule induces biological activity and has physiological consequences.

Amino Acid Sequence↗
PubMed1994

Differential RNA regulation by staphylococcal enterotoxins A and B in murine macrophages.

Staphylococcal enterotoxin A (SEA) is significantly better than enterotoxin B (SEB) in activating tumor necrosis factor (TNF) secretion by B6MP102 cells. Both toxins bound to B6MP102 cells; however, SEB competed less effectively with SEA than SEA competed with SEB. This suggested that receptors unique to SEA were present on B6MP102 cells. Signal transduction occurred in response to both toxins. Within 30 s after addition, SEA and SEB significantly increased the F-actin concentration in B6MP102 cells. However, only SEA induced increased TNF mRNA levels. B6MP102 cells incubated with interferon-gamma and SEB secreted TNF. However, enhanced mRNA expression was delayed and the concentration of TNF secreted was less than that of B6MP102 cells stimulated with SEA. Although these data suggest that receptors unique to SEA are present on B6MP102 cells, they also indicate that staphylococcal enterotoxins differentially regulate TNF at the RNA level, perhaps because of differences in binding to the plasma membrane.

Actins↗
PubMed1994

Class I and class II major histocompatibility molecules play a role in bone marrow-derived macrophage development.

Class I and class II major histocompatibility complex (MHC) molecules play significant roles in T cell development and immune function. We show that MHCI- and MHCII-deficient mice have low numbers of macrophage precursors and circulating monocytes, as well as abnormal bone marrow cell colony-stimulating factor type 1 secretion and bone composition. We suggest that MHCI and MHCII molecules play a significant role in macrophage development.

Animals↗
PubMed1994

Binding and activation of major histocompatibility complex class II-deficient macrophages by staphylococcal exotoxins.

Macrophages from C2D transgenic mice deficient in the expression of major histocompatibility complex (MHC) class II proteins were used to identify binding sites for superantigens distinct from the MHC class II molecule. Iodinated staphylococcal enterotoxins A and B (SEA and SEB) and exfoliative toxins A and B (ETA and ETB) bound to C2D macrophages in a concentration-dependent and competitive manner. All four toxins increased F-actin concentration within 30 s of their addition to C2D macrophages, indicating that signal transduction occurred in response to toxin in the absence of class II MHC. Furthermore, ETA, ETB, SEA, and, to a lesser extent, SEB induced C2D macrophages to produce interleukin 6. Several molecular species on C2D macrophages with molecular masses of 140, 97, 61, 52, 43, and 37 kDa bound SEA in immunoprecipitation experiments. These data indicate the presence of novel, functionally active toxin binding sites on murine macrophages distinct from MHC class II molecules.

Actins↗
PubMed1993

Staphylococcus-mediated T-cell activation and spontaneous natural killer cell activity in the absence of major histocompatibility complex class II molecules.

We used major histocompatibility complex class II antigen-deficient transgenic mice to show that in vitro natural killer cell cytotoxicity and T-cell activation by staphylococcal exotoxins (superantigens) are not dependent upon the presence of major histocompatibility complex class II molecules. T cells can be activated by exotoxins in the presence of exogenously added interleukin 1 or 2 or in the presence of specific antibody without exogenously added cytokines.

Animals↗
PubMed1993

Effect of Aspergillus oryzae fermentation extract (Amaferm) on in vitro fiber degradation.

The influence of Aspergillus oryzae fermentation extract (Amaferm) on in vitro fiber degradation was determined by incubating eight ground fibrous feed-stuffs with rumen fluid and buffer inoculum. Amaferm was added at 0, .4, .8, or 1.2 g/L of fermentation mixture. Both NDF and ADF degradabilities were determined after 96 h of incubation. Addition of extract had no effect on NDF or ADF degradability of pure cellulose, low endophyte fescue, wheat straw, corn silage, or prairie hay. Addition of Amaferm at .8 or 1.2 g/L increased NDF and ADF degradations of bromegrass hay and alfalfa hay; its addition at .4 or .8 g/L, but not at 1.2 g/L, increased NDF and ADF degradation of high endophyte fescue hay. In a second set of in vitro fermentations, selective antimicrobials (penicillin, streptomycin, and cycloheximide) were used to assess the influence of Amaferm on various microbial groups. The enhanced fiber degradation by Amaferm was attributed to its stimulation of bacterial activity because its addition to whole rumen fluid without or with cycloheximide increased fiber digestion. In contrast, addition of Amaferm to the whole rumen fluid plus penicillin and streptomycin treatment had no effect on fiber degradation, suggesting that fungal or protozoal activity was not affected by treatment. In conclusion, Amaferm increased fiber digestibility of certain feedstuffs, and the increase was mediated via stimulation of rumen bacterial, but not fungal or protozoal, activities.

Animal Feed↗
PubMed1992

Moderation of ruminal fermentation by ciliated protozoa in cattle fed a high-grain diet.

The objective of this study was to assess the influence of ciliated protozoa on ruminal fermentation in cattle fed high-grain diets. Six ruminally cannulated steers fed a corn-based grain diet (85% concentrate plus 15% alfalfa hay) at 12-h intervals were assigned randomly to two groups, ciliate free and faunated, in a crossover design. Defaunation was by ruminal emptying, omasal flushing, and treatment with sodium sulfosuccinate. Two to 3 weeks after defaunation, the ruminal contents of all steers were sampled before the morning feeding (0 h) and at 1, 2, 4, 6, 8, and 12 h after feeding to measure pH, analyze fermentation products, and monitor counts of ciliated protozoa and lactic acid-producing and -fermenting bacterial groups. Total numbers of ciliated protozoa in the faunated steers averaged 4.3 x 10(5)/g, and the protozoa consisted of nine genera. Ciliate-free steers had lower (P less than 0.01) ruminal pHs (pH 5.97) than faunated cattle (pH 6.45); however, the treatment-time interaction was not significant. Ruminal lactate and ammonia concentrations were similar in both groups. The total volatile fatty acid concentration was higher (P less than 0.05) in the ciliate-free steers than in the faunated steers and exhibited a treatment-time interaction (P less than 0.05). The acetate-to-propionate ratio was higher (P less than 0.05) in the faunated group than in the ciliate-free group and showed a treatment-time interaction (P less than 0.05). Total anaerobic bacterial counts were about fourfold higher in the ciliate-free group than in the faunated group.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

Performance and ruminal function development of young calves fed diets with Aspergillus oryzae fermentation extract.

Neonatal Holstein heifer (n = 72) and bull (n = 40) calves were used to study the effects of Aspergillus oryzae fermentation extract (Amaferm) on their performance and on rumen development. The starter diets were formulated to achieve Amaferm consumption of 0, .5, 1, or 3 g per calf daily. Calves were fed milk daily and allowed to consume starter and a mixture of alfalfa and bromegrass hay ad libitum. Weaning was when calves consumed 550 g of starter on 2 consecutive d. Weight gain and feed consumption were recorded weekly. Forty of the heifer calves, 10 from each treatment, were selected randomly to study the effects of Amaferm on ruminal fermentative development. Ruminal fluid samples were collected for pH, ruminal fermentation products, and for bacterial enumerations. Overall, Amaferm-supplemented calves were weaned 1 wk earlier than unsupplemented calves. They had higher total VFA, propionate, and acetate concentrations in the rumen than unsupplemented calves. Total anaerobic, hemicellulolytic, and pectinolytic bacterial counts were higher; cellulolytic bacterial counts tended to be higher for the Amaferm-supplemented calves than for controls. In general, Amaferm-supplemented calves had greater ruminal microbial activity than those not fed Amaferm.

Animal Feed↗
PubMed1990

Supplementation of dormant tallgrass-prairie forage: I. Influence of varying supplemental protein and(or) energy levels on forage utilization characteristics of beef steers in confinement.

Three experiments were conducted to evaluate effects of supplemental protein vs energy level on dormant forage intake and utilization. In Exp. 1, 16 ruminally cannulated steers were blocked by weight (avg wt = 242 kg) and assigned randomly to a negative control or to one of three isocaloric supplement treatments fed at .4% BW: 1) control, no supplement (NS); 2) 12% CP, low protein (LP); 3) 28% CP, moderate protein (MP); 4) 41% CP, high protein (HP). In Exp. 2 and 3, 16 ruminally cannulated steers were blocked by weight (avg wt = 332 kg, Exp. 2; 401 kg, Exp. 3) and assigned randomly to a 2 x 2 factorial arrangement of treatments. The treatments contrasted low (LP) and high (HP) levels of supplemental protein (.66 g CP/kg BW vs 1.32 g CP/kg BW) with low (LE) and high (HE) levels of supplemental ME (9.2 kcal/kg BW vs 18.4 kcal/kg BW). In Exp. 1, forage DMI as well as ruminal DM and indigestible ADF fill at 4 h postfeeding were greater (P less than .10) with the MP and HP steers than with control and LP steers. Total DM digestibility increased (P less than .10) for supplemented steers (35.5% for control vs 47.3 for supplemented steers); however, LP depressed (P less than .10) NDF digestibility. In Exp. 2, forage DMI, indigestible ADF flow and liquid flow were depressed (P less than .10) in LP-HE supplemented steers. In Exp. 3, HP steers had greater (P less than .10) forage DMI, indigestible ADF fill values (4 h postfeeding), liquid volume and tended (P = .11) to have greater ruminal DM fill (4 h postfeeding). In summary, increased levels of supplemental protein increased intake and utilization of dormant tallgrass-prairie forage (less than 3% CP). Increasing supplemental energy without adequate protein availability was associated with depressed intake and digestibility.

Animal Feed↗