Activation of bovine neutrophils by recombinant bovine tumor necrosis factor-alpha

The in vitro effect of bovine recombinant tumor necrosis factor-alpha (rbTNF-alpha) on bovine neutrophil function and the possibility that rbTNF-alpha and recombinant bovine interferon-gamma (rbIFN-gamma) act synergistically were investigated. Treatment of neutrophils with rbTNF-alpha (0.05 micrograms/ml; approximately 50 U/ml) at 37 degrees C for 2.5 h resulted in enhancement of antibody independent neutrophil-mediated cytotoxicity (AINC) and inhibition of random migration and chemotaxis. The same treatment resulted in a slight decrease in iodination and cytochrome C reduction, but did not affect Staphylococcus aureus ingestion, or antibody dependent cell-mediated cytotoxicity. Kinetic and inhibitor studies indicated that the action of rbTNF-alpha was rapid and was independent of protein and RNA synthesis by neutrophils. Evaluation of the synergistic activities of rbTNF-alpha and rbIFN-gamma indicated that treatment of neutrophils with these two cytokines simultaneously resulted in additive enhancement of AINC and inhibition of random migration and chemotaxis. There was no additive effect of the two cytokines on inhibition of iodination or cytochrome C reduction.     

Effects of vitamins A and E on superoxide production and intracellular signaling of neutrophils in Holstein calves.

The effects of vitamin A and vitamin E treatment on superoxide (O2-) production of neutrophils and their intracellular signaling, including intracellular Ca2+, protein kinase C (PKC), and tyrosine kinase (TK), were examined in Holstein calves. After treatment with vitamin A, heat-aggregated IgG (H-agg.IgG)-induced O2- production in neutrophils ranged from 3.7+/-0.4 to 4.3+/-0.9 nmol (mean +/- SD), and it was significantly (P<0.05) higher than that in the control calves. Opsonized zymosan (OPZ)-induced O2- production was similar with that of control calves. Intracellular signaling of neutrophils in vitamin A-treated calves was enhanced compared with that of control calves when stimulated with H-agg. IgG, but not with OPZ. After treatment with vitamin E, H-agg.IgG- and OPZ-induced O2- production of neutrophils ranged from 4.2+/-0.8 nmol to 5.4+/-0.5 nmol and from 7.8+/-0.6 nmol to 8.1+/-0.4 nmol (mean +/- SD), respectively, and they were significantly (P<0.05) higher than those in control calves. Intracellular signaling was also enhanced compared with that of control calves. These results suggested that the effects of vitamin A and E treatment on O2- production were correlated to changes in intracellular signaling of neutrophils in Holstein calves.     

Plasma leptin responses to lipopolysaccharide and tumor necrosis factor alpha in cows

Peripheral administration of bacterial lipopolysaccharide (LPS) and various inflammatory cytokines to rodents is known to raise plasma levels of leptin, a potent satiety factor secreted from adipocytes, implying a role of leptin in endotoxin-induced anorexia. We previously reported no effect of LPS on serum leptin levels in sheep, despite marked anorexia and fever. Our results suggest that leptin might not be involved in the endotoxin-induced anorexia in ruminants. To test this idea, in the present study, plasma leptin levels were measured during acute experimental endotoxemia in Holstein cows. Intravenous injection of LPS induced anorexia accompanied with increases in plasma levels of cortisol and insulin, all of which are known to stimulate leptin secretion in rodent and human, while it did not affect plasma leptin levels at all in cows. Similar results were also obtained after injection of recombinant bovine tumor necrosis factor alpha. These results indicate that plasma leptin levels in cows during acute endotoxemia are differentially regulated from those in rodents, and that leptin might not be involved in the endotoxin-induced anorexia in ruminants.     

Effects of diet energy density and milking frequency in early lactation on tumor necrosis factor-alpha responsiveness in dairy cows

A whole blood stimulation assay (WBA) with Escherichia coli lipopolysaccharide (LPS) and an enzyme-linked immunosorbent assay (ELISA) were established to measure the production of tumor necrosis factor-alpha (TNF-alpha) in bovine plasma. The assays were used to study the effect of time around parturition, and diet energy density, and milking frequency on TNF-alpha responsiveness of dairy cows in early lactation. Forty cows were included in a 2 x 2 factorial block design. One factor was high (H) versus low (L) diet energy density and the other factor was two versus three daily milkings. Blood samples were collected in weeks -3, -1, 2, 3, 5, 9, and 13 around parturition, and investigated for the TNF-alpha production ex vivo and CD14+ monocytes. The TNF-alpha response, CD14+ monocyte number, and CD14 expression level on monocytes were significantly increased in the weeks close to parturition. However, dips of varying sizes were observed for the measured parameters in week 3 after calving. Diet and milking frequency had no effect on the TNF-alpha response ex vivo or CD14 expression level on monocytes, but cows fed diet H had significantly higher numbers of CD14+ monocytes than cows fed diet L. The WBA with LPS was a fast reliable method for repeated measurements of TNF-alpha responsiveness in cattle. Previous findings of increased TNF-alpha responses in periparturient cows were confirmed, whereas diet energy concentration and milking frequency had no effect on the TNF-alpha responsiveness in early lactation.     

外源性金属硫蛋白对中国荷斯坦奶牛产能性能和内分泌的影响

选取24头泌乳奶牛,随机分成A、B、C和D,4组,分别按剂量0.0(对照),6.0,12.0和16.0 mg/头静脉注射金属硫蛋白(MT),以探讨外源性MT对奶牛产能性能和内分泌的影响.结果表明,B组1～30 d、C组1～45 d的标准产奶量显著高于A组,C组乳脂率在31～45 d较A组有显著改善;B组INS含量30 ...     

复合饲料添加剂对荷斯坦奶牛生产性能及血清生化指标的影响

文章旨在探究复合型饲料添加剂对荷斯坦奶牛生产性能、血清生化指标的影响.采取单因素试验设计,将18头身体健康、体况相近的泌乳中期荷斯坦奶牛随机分为2组,分别为对照组、试验组(添加复合型饲料200?g/d),其中每组设置3个重复,每个重复3头牛,整个试验周期为30?d.于试验期第15和30天采集奶牛乳样及空腹血样,用于测定...     

Effect of boron supplementation of pig diets on the production of tumor necrosis factor-alpha and interferon-gamma

Two experiments were conducted to determine the effects of dietary B on the production of cytokines following an endotoxin challenge. In both experiments, pigs were obtained from litters generated from sows fed low-B (control) or B-supplemented (5 mg/ kg, as-fed basis) diets. In Exp. 1 and 2, 28 and 35 pigs, respectively (21 d old), remained with their littermates throughout a 49-d nursery phase and were fed either a control or B-supplemented diet. In Exp. 1, 12 pigs per treatment were moved to individual pens at the completion of the nursery phase and fed their respective experimental diet. On d 99 of the study, pigs were injected with 150 microg of phytohemagglutinin (PHA) to evaluate a local inflammatory response. Pigs receiving the B-supplemented diet had a decreased (P < 0.01) inflammatory response following PHA injection. Peripheral blood monocytes were isolated from six pigs per treatment on d 103 and cultured in the presence of lipopolysaccharide (LPS) to determine the effect of dietary B on tumor necrosis factor-alpha (TNF-alpha) production from monocytes. Isolated monocytes from pigs that received the B-supplemented diet had a numerically greater (P = 0.23) production of TNF-alpha. In Exp. 2, pigs were group housed with their littermates following the nursery phase for 43 d, after which 10 pigs per treatment were moved to individual pens. In Exp. 1 and 2, pigs were assigned randomly within dietary treatment to receive either an i.m. injection of saline or LPS on d 117 and d 109, respectively. The dose of LPS in Exp. 1 and 2 was 100 and 25 microg of LPS/kg of BW, respectively. In Exp. 1, serum TNF-alpha was increased (P < 0.01) at 2 h and tended to be increased (P < 0.11) at 6 and 24 h after injection by dietary B; however, only numerical trends existed for a B-induced increase in TNF-alpha in Exp. 2. Serum interferon-gamma (IFN-gamma) was increased (P < 0.01) at 6 h and tended to be increased (P < 0.08) at 24 h after injection in Exp. 1. In Exp. 2, dietary B also numerically increased IFN-alpha. These data indicate that dietary B supplementation increased the production of cytokines following a stress, which indicates a role of B in the immune system; however, these data do not explain the reduction in localized inflammation following an antigen challenge in pigs.     

Effects of nitric oxide and tumor necrosis factor-alpha on production of prostaglandin F2alpha and E2 in bovine endometrial cells

The purpose of this study was to determine whether nitric oxide (NO) mediates tumor necrosis factor (TNF)alpha influence on the bovine endometrium. TNFalpha influence on the bovine endometrium is limited to the stromal cells. Therefore, it was interesting to find out whether NO production by the stromal cells, stimulated by TNFalpha might influence the endometrial epithelium. Moreover, we investigated the intracellular mechanisms of TNFalpha- and NO-regulated prostaglandin (PG) F(2alpha) and PGE(2) synthesis. Epithelial and stromal cells from the bovine endometrium (Days 2-5 of the oestrous cycle) were separated by means of enzymatic dispersion and cultured for 6-7 days in 48-well plates. The confluent endometrial cells were exposed to a NO donor (S-NAP; 1-1000 microM) for 24 h. S-NAP strongly stimulated PGE(2) production in both bovine endometrial cell types (P<0.001). The effect of SNAP on PGF(2alpha) production was limited only to the stromal cells (P<0.05). To study the intracellular mechanisms of TNFalpha and NO action, stromal cells were incubated for 24 h with TNFalpha or S-NAP and with NO synthase (NOS) inhibitor (L-NAME; 10 microM) or an inhibitor of phosphodiesterase (IBMX; 10 microM). When the cells were exposed to TNFalpha in combination with NOS inhibitor (L-NAME), TNFalpha-stimulated PGs production was reduced (P<0.05). The inhibition of enzymatic degradation of cGMP by IBMX augmented the actions of S-NAP and TNFalpha on PGs production (P<0.05). The overall results suggest that TNFalpha augments PGs production by bovine endometrial stromal cells partially via induction of NOS with subsequent stimulation of NO-cGMP formation. NO also stimulates PGE(2) production in epithelial cells.     

不同能量摄入水平对围产期奶牛生产性能及血浆瘦素浓度影响的研究

将健康的围产期奶牛30头随机分为3组, 每组10头。其中Ⅱ组为对照组, Ⅰ组和Ⅲ组分别为试验组。Ⅰ、Ⅱ、Ⅲ组均于产前28d开始分别饲喂NRC标准减少20%日粮(能量摄入80%组)、NRC标准日粮(能量摄入100%组)、NRC标准增加20%日粮(能量摄入120%组), 产后各组奶牛均饲喂标准日粮, 至产后56d结束。观察妊娠后期奶牛不同能量水平对干物质摄入量、产奶量、乳成分及体重等生产性能和血浆瘦素浓度的影响。试验结果表明: 奶牛产后干物质摄入量、产奶量、乳中蛋白质、乳脂率和非固体物质为Ⅰ组高于Ⅱ组、Ⅱ组又高于Ⅲ组(P>0 05 ); 同时血浆瘦素浓度为Ⅰ组高于Ⅱ组、Ⅱ组又高于Ⅲ组, 经统计分析组间差异极显著(P<0 01) 或差异显著(P<0 05); 但产前28~14d瘦素浓度组间差异均不显著(P>0 05 ), 此后Ⅰ组和Ⅲ组差异显著(P<0 05 )。此外, Ⅰ组体重消耗最小, Ⅲ组体重消耗最大, 各组间差异极显著(P<0 01)。可见, 围产期健康奶牛不同能量水平与干物质摄入量、产奶量、乳成分、体重等生产性能及血浆瘦素浓度之间存在着密切的关系。     

Evaluation of phagocytosis, bactericidal activity, and production of superoxide anion, nitric oxide, and tumor necrosis factor-alpha in Kupffer cells of neonatal pigs.

OBJECTIVE: To evaluate the activity of Kupffer cells (KC) of control neonatal pigs and neonatal pigs treated with endotoxin and to compare activity of KC with that of pulmonary alveolar macrophages (PAM). SAMPLE POPULATION: Kupffer cells and PAM obtained from 24 neonatal pigs (7 to 10 days old). PROCEDURE: Pairs (n = 7) of littermates served as treated (lipopolysaccharide [LPS]) or untreated pigs. Pigs were euthanatized 24 hours after treatment, and cells were isolated. Cells were obtained from 10 other neonatal pigs for other assays. Functional activity of cells was evaluated by use of in vitro assays to evaluate bactericidal activity, phagocytosis, and production of superoxide anion (SOA), nitric oxide (NO), and tumor necrosis factor-alpha (TNF-alpha). Each assay was repeated on cells obtained from 4 to 6 pigs. RESULTS: Phagocytic activity was similar in KC and PAM, but bactericidal activity and production of SDA and TNF-alpha was lower in KC. Neither KC nor PAM produced NO in response to LPS stimulation. Phagocytosis, bactericidal activity, and production of SOA were enhanced for KC obtained from neonatal pigs treated with LPS. The PAM from LPS-treated neonatal pigs had similar bactericidal activity to PAM obtained from untreated pigs. CONCLUSIONS AND CLINICAL RELEVANCE: Functional capacity of KC is affected by endotoxin. This provides additional information of the role the liver plays in immune surveillance. In addition, the response of KC in neonatal pigs exposed to endotoxin is of value for understanding gram-negative bacterial sepsis, which is a major cause of mortality in neonatal pigs.     

Possible actions of tumor necrosis factor-alpha in ovarian function

Tumor necrosis factor-alpha (TNFalpha) is a multifunctional cytokine that was first described as a tumoricidal factor produced by activated macrophages. Extensive research over the last two decades has suggested that TNFalpha has physiologically diverse actions in ovarian function in a variety of species. TNFalpha and its specific receptors are present in the ovaries of many species. Furthermore, TNFalpha plays multiple and probably important roles in corpus luteum (CL) function as well as ovarian cell function throughout the estrous cycle. This review focuses on recent studies documenting TNFalpha in ovarian follicles and CL in several mammals. In addition, possible roles of TNFalpha in ovarian function throughout the estrous cycle and in the gestation period are discussed.     

Effects of tissue inhibitor of metalloproteinases on canine chondrocytes cultured in vitro with tumor necrosis factor-alpha

OBJECTIVE: To elucidate tissue inhibitor of metalloproteinase (TIMP)-mediated effects on chondrocytes. SAMPLE POPULATION: Articular cartilage from humeral heads of 6 dogs. PROCEDURE: Chondrocytes from harvested specimens were cultured in 3-dimensional (3-D) agarose at 10(6) cells/mL. We prepared 3-D constructs exposed to only tumor necrosis factor (TNF)-alpha (50 ng/mL). Recombinant human TIMP-1 (255nM), -2 (285nM), or -3 (250nM) was added to liquid media bathing 3-D constructs cultured with TNF-alpha. Chondrocytes cultured without TIMP or TNF-alpha served as control samples. Samples of liquid media were collected on days 6, 9, 15, and 21 of culture for evaluation of glycosaminoglycan (GAG) and nitric oxide concentrations. The 3-D constructs were collected on days 9, 15, and 21 for evaluation of GAG, hydroxyproline (HP), and DNA contents. RESULTS: GAG content in control samples increased significantly during the study, whereas GAG content in 3-D constructs cultured with TNF-alpha or TNF-alpha plus TIMP did not increase. On day 9, GAG release from 3-D constructs cultured with TNF-alpha was significantly higher than that in other constructs. The HP content in control samples increased during the study and was significantly higher than that in all other constructs on day 21. Concentrations of nitric oxide were significantly lower in control samples on day 6, compared with concentrations for all other constructs. CONCLUSIONS AND CLINICAL RELEVANCE: Addition of TIMPs did not counteract suppression of GAG and HP accumulation in 3-D constructs exposed to TNF-alpha. Apparently, adverse effects on chondrocytes exposed to TNF-alpha cannot be prevented by addition of TIMP alone.     

Adenosine A2A receptor agonists inhibit lipopolysaccharide-induced production of tumor necrosis factor-alpha by equine monocytes

Adenosine is an endogenous nucleoside that regulates many physiological processes by activating one or more adenosine receptor subtypes, namely A1, A2A, A2B and A3. The results of previous studies indicate that adenosine analogues inhibit lipopolysaccharide (LPS)-induced production of reactive oxygen species (ROS) by equine neutrophils primarily through activation of A2A receptors. Because peripheral blood monocytes produce cytokines that are responsible for many of the deleterious effects of LPS, the current study was performed to evaluate the effects of an array of novel adenosine receptor agonists on LPS-induced production of tumor necrosis factor-alpha (TNF-alpha), and to assess the selectively of these agonists for equine adenosine A2A over the A1 receptor. Radioligand binding studies performed with equine tissues expressing adenosine A1 and A2A receptor subtypes yielded a rank order of affinity for the equine A2A receptor of ATL307>ATL309 approximately ATL310 approximately ATL313>ATL202 approximately ATL361 approximately ATL376>ATL372>CGS21680>NECA. Co-incubation of equine peripheral blood monocytes with LPS and these agonists resulted in inhibition of TNF-alpha production with a rank order of potency that strongly correlated with their binding affinities for equine adenosine A2A receptors. Results of experiments performed with one of the adenosine receptor agonists (ATL313) and selective adenosine receptor antagonists confirmed that inhibition of LPS-induced production of TNF-alpha occurred via stimulation of A2A receptors. Although incubation of monocytes with IB-MECA, a compound purported to act as an adenosine A3 receptor agonist, reduced LPS-induced TNF-alpha production, this effect of IB-MECA was inhibited by the A2A selective antagonist ZM241385 but not by the A3 receptor antagonist MRS1220. These results indicate that the adenosine receptor subtype responsible for regulation of LPS-induced cytokine production by equine monocytes is the A2A receptor. To address the signal transduction mechanism responsible for the anti-inflammatory effects of ATL313 in equine monocytes, production of cAMP was compared in the presence and absence of either the adenosine A2A receptor antagonist ZM241385 or the adenosine A2B receptor antagonist MRS1706. In the absence of the antagonists, ATL313 increased production of cAMP; ZM241385 inhibited this effect of ATL313, whereas MRS1706 did not. Furthermore, incubation of monocytes with either the stable analogue of cAMP, dibutyryl cAMP, or forskolin, an activator of adenylyl cyclase, also inhibited LPS-induced production of TNF-alpha production by equine monocytes. Collectively, the results of the current study indicate that adenosine analogues inhibit LPS-induced production of TNF-alpha by equine monocytes primarily via activation of adenosine A2A receptors and do so in a cAMP-dependent manner. The results of this study indicate that stable adenosine analogues that are selective for adenosine A2A receptors may be suitable for development as anti-inflammatory drugs in horses.     

Adrenocortical responses to tumor necrosis factor-alpha and interferon-gamma in cattle

The responses of plasma cortisol and adrenocorticotropic hormone (ACTH) were examined to intravenous injection of recombinant bovine tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (INF-gamma) in Holstein cows. INF-gamma induced dose-dependent rises in the plasma levels of both cortisol and ACTH, while TNF-alpha induced comparable plasma cortisol responses with much smaller rises in plasma ACTH. The results suggest a direct stimulatory action of TNF-alpha on cortisol secretion from the adrenal gland in cattle.     

Effects of high ambient temperature and restricted feed intake on nitrogen utilization for milk production in lactating Holstein cows

Three experiments were performed to examine the effects of high ambient temperature and feed restriction on nitrogen (N) utilization in lactating cows. Experiment 1 investigated N utilization in four cows fed ad libitum in a 2 × 2 crossover design under constant moderate (18°C) or high (28°C) ambient temperatures. The milk N secretion (P < 0.01) and protein concentration (P < 0.05) decreased under high ambient temperature. Experiment 2 investigated N utilization in four cows under constant moderate ambient temperature in a 2 × 2 crossover design with ad libitum or 70% ad libitum feed intake. The milk N secretion and protein concentration both decreased with feed restriction (P < 0.05). Experiment 3 investigated N utilization in four cows fed 70% ad libitum in a 2 × 2 crossover design under constant moderate or high ambient temperatures. The milk protein concentration decreased under high ambient temperature (P < 0.01). The milk N secretion tended to decrease under high ambient temperature (P < 0.10). Therefore, decreased N utilization for milk production at high ambient temperature is mainly caused by a reduced feed intake and the high ambient temperature itself.     

荷斯坦奶牛体型线性性状与产奶量的相关性分析

对新疆荷斯坦奶牛的产奶量与体型线性性状进行了相关。通径分析。结果表明，荷斯坦奶牛的乳房纵沟深，后乳房宽，后乳房高与产奶量呈中等正相关，体高，体长，尻宽，尻长与产奶量呈弱相关；后乳房宽，乳房纵沟深是直接影响产奶量的重要性状，且是正身作用；后乳房高对产奶量起负面影响。其间接作用大。     

Phagocytosis and killing of Staphylococcus aureus by bovine neutrophils after priming by tumor necrosis factor-alpha and the des-arginine derivative of C5a

OBJECTIVES: To evaluate effects of proinflammatory mediators on phagocytosis and killing of Staphylococcus aureus, the oxidative burst (OB), and expression of receptors for opsonins by bovine neutrophils. SAMPLE POPULATION: Neutrophils from 10 cattle. PROCEDURE: Neutrophils were primed with recombinant bovine tumor necrosis factor-alpha (TNF-alpha) or the des-arginine derivative of bovine C5a (C5a(desArg)) and mixed with S aureus. Phagocytosis and OB were measured by use of flow cytometry. Rate of phagocytosis and intracellular killing were evaluated. Expression of receptors for immunoglobulins and the C3bi fragment of complement were estimated by use of flow cytometry. RESULTS: Priming of neutrophils by TNF-alpha improved phagocytosis of S aureus with a concentration-dependent effect. Phagocytosis of preopsonized washed bacteria was increased by activation of neutrophils with C5a(desArg). Phagocytosis was optimal when neutrophils primed with TNF-alpha were activated with C5a(desArg). The OB of phagocytizing neutrophils was highest when TNF-alpha and C5a(desArg) were used in combination. Bactericidal activity of neutrophils was stimulated by priming with TNF-alpha or C5a(desArg). Binding of bovine IgM or IgG2 to bovine neutrophils was not stimulated byTNF-alpha, C5a(desArg), or both, and aggregated IgG1 did not bind to neutrophils regardless of their activation state. Both TNF-alpha and C5a(desArg) increased expression of beta2 integrins (CD18), with the highest expression when they were used in combination. CONCLUSIONS AND CLINICAL RELEVANCE: The mediators TNF-alpha and C5a(desArg) stimulated phagocytic killing by neutrophils and potentiated each other when used at suboptimal concentrations. Bovine neutrophils have enhanced bactericidal activities at inflammatory sites when TNF-alpha, C5a(desArg), or both are produced locally.     

Effects of long-term administration of recombinant bovine tumor necrosis factor-alpha on glucose metabolism and growth hormone secretion in steers

OBJECTIVE: To investigate the effects of long-term administration of recombinant bovine tumor necrosis factor-alpha (rbTNF) on plasma glucose and growth hormone concentrations, and to determine whether treatment with rbTNF causes insulin resistance in steers. ANIMALS: 5 steers treated with rbTNF and 5 steers treated with saline (0.9% NaCl) solution (control). PROCEDURES: In experiment 1, rbTNF (5.0 microg/kg of body weight) or saline solution (5 ml) was administered SC daily for 12 days. Blood samples were obtained before treatment, and plasma was harvested for determination of glucose, insulin, and growth hormone (GH) concentrations. In experiment 2, insulin, glucose, or growth hormone-releasing hormone (GHRH) was administered IV on days 7, 9, and 11, respectively, after initiation of rbTNF or saline treatment in experiment 1. Plasma glucose and insulin concentrations were measured before and at various times for 4 hours after insulin or glucose administration. Plasma GH concentrations were measured at various times for 3 hours after GHRH administration. RESULTS: In experiment 1, administration of rbTNF resulted in hyperinsulinemia without hypoglycemia and decreased plasma GH concentrations. In experiment 2, plasma glucose concentrations were higher in steers treated with rbTNF and insulin than in controls. Plasma GH concentrations were lower in steers treated with rbTNF and GHRH than in controls. CONCLUSIONS AND CLINICAL RELEVANCE: Prolonged treatment with rbTNF induced insulin resistance and inhibited GHRH-stimulated release of GH in steers. Results indicate that rbTNF is a proximal mediator of insulin resistance and inhibits release of GH during periods of endotoxemia or infection.     

Effects of interleukin-1beta and tumor necrosis factor-alpha on expression of matrix-related genes by cultured equine articular chondrocytes

OBJECTIVE: To determine the effects of interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) on expression and regulation of several matrix-related genes by equine articular chondrocytes. SAMPLE POPULATION: Articular cartilage harvested from grossly normal joints of 8 foals, 6 yearling horses, and 8 adult horses. PROCEDURE: Chondrocytes maintained in suspension cultures were treated with various doses of human recombinant IL-1beta or TNF-alpha. Northern blots of total RNA from untreated and treated chondrocytes were probed with equine complementary DNA (cDNA) probes for cartilage matrix-related genes. Incorporation of 35S-sulfate, fluorography of 14C-proline labeled medium, zymography, and western blotting were used to confirm effects on protein synthesis. RESULTS: IL-1beta and TNF-alpha increased steady-state amounts of mRNA of matrix metalloproteinases 1, 3, and 13 by up to 100-fold. Amount of mRNA of tissue inhibitor of metalloproteinase-1 also increased but to a lesser extent (1.5- to 2-fold). Amounts of mRNA of type-II collagen and link protein were consistently decreased in a dose-dependent manner. Amount of aggrecan mRNA was decreased slightly; amounts of biglycan and decorin mRNA were minimally affected. CONCLUSIONS AND CLINICAL RELEVANCE: Treatment of cultured equine chondrocytes with IL-1beta or TNF-alpha resulted in marked alterations in expression of various matrix and matrix-related genes consistent with the implicated involvement of these genes in arthritis. Expression of matrix metalloproteinases was increased far more than expression of their putative endogenous inhibitor. Results support the suggestion that IL-1beta and TNF-alpha play a role in the degradation of articular cartilage in arthritis.     

日粮添加碳酸氢钠对荷斯坦奶牛生产性能的影响

选择体重、胎次、泌乳期、乳脂率和产奶量基本一致的健康无病的中国荷斯坦奶牛30头,随机分成3组,分别为试验组Ⅰ,Ⅱ和对照组。试验组Ⅰ,Ⅱ奶牛日粮每日每头分别添加130、150g碳酸氢钠,对照组奶牛日粮每日每头分别添加110g碳酸氢钠,试验期28d。结果表明,试验组Ⅰ干物质采食量、粪便pH比对照组高1.41%、0.20(P0.05),非脂乳固体比对照组低0.4(P0.05);试验组Ⅱ干物质采食量、产奶量、4%标准乳、粪便pH分别比对照组高2.04%、2.01%、2.68%、0.42(P0.05),乳脂率高0.13%(P0.05),乳蛋白、非乳脂固体分别低0.36%、0.51%(P0.05)。另外,碳酸氢钠的增加使得牛乳中体细胞数显著降低,且与乳脂肪呈显著负相关(P0.05),而与乳蛋白质呈显著正相关(P0.05)。由此可知,在每头奶牛日粮中添加150g碳酸氢钠的范围内,随着日粮中碳酸氢钠的增加,能显著改善荷斯坦奶牛的生产性能,有效预防奶牛瘤胃酸中毒。