In vitro expression and inhibition of procoagulant activity produced by bovine alveolar macrophages and peripheral blood cells

Local and systemic activation of coagulation is frequently associated with bacterial sepsis. The coagulopathy is due, at least in part, to expression of tissue factor (TF) by monocytes and macrophages. The purpose of this study was to evaluate the expression of procoagulant activity by bovine alveolar macrophages, leukocytes and platelets, and to determine the relative potency of three chemical inhibitors of TF expression (pentoxifylline, retinoic acid, and cyclosporin A). Bovine alveolar macrophages were stimulated with lipopolysaccharide (LPS) derived from Pasteurella haemolytica or recombinant bovine tumour nervous factor (TNF) and dose- and time-dependent effects on TF expression were studied. LPS and TNF induced TF expression in alveolar macrophages and LPS treatment of whole blood induced TF expression in mononuclear cells. Neutrophils and platelets also expressed procoagulant activity, but this activity was not inhibited by anti-bovine TF monoclonal antibody. Pentoxifylline (40 mol/L), retinoic acid (0.01 mmol/L) and cyclosporin A (0.08 mol/L) inhibited TF expression when added concurrently with LPS or TNF, but not when added 4 h after stimulation. TF mRNA was not detected in unstimulated alveolar macrophages by Northern blot analysis. In contrast, exposure to LPS or TNF for 6 h induced marked expression of TF mRNA, which was inhibited by treatment with pentoxifylline, retinoic acid and cyclosporin A. Expression of TNF by alveolar macrophages stimulated with LPS was also inhibited by these compounds. Our results indicate that procoagulant activity expressed by alveolar macrophages and monocytes is associated with expression of TF, whereas procoagulant activity expressed by neutrophils and platelets is not. The concentrations of pentoxifylline and retinoic acid necessary for inhibition of TF expression in vitro may not be achievable in vivo owing to their toxic effects. However, the in vitro concentration of cyclosporin A that inhibited TF expression did not exceed the plasma concentration observed in humans, and therefore may be useful for inhibition of TF expression in vivo.Abbreviations BAL bronchoalveolar lavage - LPS lipopolysaccharide - cDNA cloned deoxyribonucleic acid - cAMP cyclic adenosine monophosphate - GAPDH glyceraldehyde phosphate dehydrogenase - mRNA messenger ribonucleic acid - TF tissue factor - TNF tumour necrosis factor - DPBS Dulbecco's phosphate-buffered saline     

Photosynthesis, symbiotic N and C accumulation in leaves of 30 nodulated cowpea genotypes grown in the field at Wa in the Guinea savanna of Ghana

Cowpea is a major food legume in Africa with relatively little information on N and C nutrition under field conditions in the continent. The aim of this study was to assess the relationship between leaf symbiotic N nutrition and photosynthetic carbon assimilation in cowpea genotypes grown under field conditions in the Guinea savanna of Ghana. The experiments were conducted in a randomized complete block design in Ghana, using 30 nodulated cowpea genotypes. Data collected from gas-exchange studies showed that genotypes with high photosynthetic rates expectedly exhibited greater stomatal conductance, high transpiration rates and increased water-use efficiency, leading to greater dry matter yield. In contrast, those with low photosynthetic rates revealed low stomatal conductance, low transpiration rates and low water-use efficiency. Quantification of symbiotic N in photosynthetic leaves indicated greater N₂ fixation in genotypes with higher photosynthetic rates, leaf transpiration and better water-use efficiency. Furthermore, measurement of C in leaves also indicated greater accumulation in genotypes with higher symbiotic N yield and total N, clearly indicating a link between C and N metabolism in photosynthetic leaves. Additionally, measures of photosynthetic N-use efficiency showed that genotypes with high levels of C-fixed per unit leaf total N also exhibited greater C-fixed per unit leaf N-fixed. Data suggest that where symbiotic legumes derive a large proportion of their N from N₂ fixation, photosynthetic C yield is much enhanced.     

Effect of nano-SiO2 on properties of wood/polymer/clay nanocomposites

Wood polymer nanocomposites (WPNC) based on nano-SiO₂ were prepared by impregnation of styrene acrylonitrile copolymer (SAN), SiO₂ nanoparticles modified with γ-trimethoxy silyl propyl methacrylate (MSMA), and nanoclay into wood. The structure of modified SiO₂ nanoparticles and WPNC was characterized by Fourier transform infrared spectroscopy (FTIR). XRD analysis showed the delaminated structure of SAN/SiO₂/clay-treated wood composites. The synergistic effect of nano-SiO₂ and nanoclay was investigated. Thermal stability of SiO₂ nanoparticles decreased after modification, while that of wood treated with SAN, SiO₂, and nanoclay improved. Morphological characteristics were examined by scanning electron microscopy (SEM). Mechanical properties, water uptake (%), dimensional stability, hardness, and flammability were found to improve due to incorporation of SiO₂ and nanoclay into wood polymer composites. Maximum improvement in properties was observed in the wood polymer composites containing SiO₂ and nanoclay at the ratio of 1:1.     

Regulation of blood glucose by hypothalamic pyruvate metabolism

The brain keenly depends on glucose for energy, and mammalians have redundant systems to control glucose production. An increase in circulating glucose inhibits glucose production in the liver, but this negative feedback is impaired in type 2 diabetes. Here we report that a primary increase in hypothalamic glucose levels lowers blood glucose through inhibition of glucose production in rats. The effect of glucose requires its conversion to lactate followed by stimulation of pyruvate metabolism, which leads to activation of adenosine triphosphate (ATP)-sensitive potassium channels. Thus, interventions designed to enhance the hypothalamic sensing of glucose may improve glucose homeostasis in diabetes.     

A seroepidemiologic survey of antibody to bluetongue virus in Alabama cattle

Bluetongue (BT) is an insect transmitted viral disease of sheep that often causes mild or inapparent disease but rarely causes severe disease in cattle. Until recently, bluetongue viral infection was believed to be more prevalent in the Western United States, as compared with other regions of the country. However, a national survey for bluetongue antibody and clinical evidence of the disease in the Southeastern United States prompted the present investigation that was designed to determine the serological prevalence of BT virus in Alabama cattle. Results of the study demonstrated that 16% of the samples collected from 1,500 cattle in 64 of the 67 counties were positive. The prevalence of positive cattle in the western part of the State was significantly higher (P less than .001) than the prevalence in the eastern half of the State. On a herd basis, 52% of all herds tested had positive animals. Results of this study suggest that bluetongue infection is more common in the Southeastern United States than previously suspected.     

Effects of dietary thiamin supplement on growth,body composition and intestinal enzyme activities of juvenile Jian carp (Cyprinus carpio var. Jian)

A total of 1050 juvenile Jian carp (Cyprinus carpio var. Jian) (8.20 ± 0.02 g) were fed diets containing seven graded levels of thiamin (0.25, 0.48, 0.79, 1.06, 1.37, 1.63 and 2.65 mg kg^?1) for 60 days to investigate the effects of thiamin on growth, body composition and digestive enzyme activities. Percent weight gain (PWG), feed intake and feed efficiency (FE) were the lowest in fish fed the basal diet (P < 0.05). Protein productive value and lipid productive value increased with increasing dietary thiamin levels up to 1.06 and 0.79 mg kg^?1 diet, respectively (P < 0.05). Body protein and lipid increased with increasing dietary thiamin levels (P < 0.05), while moisture and ash of fish carcasses decreased with the increase in dietary thiamin supplementation (P < 0.05). Intestinal folds height had a similar trend to PWG (P < 0.05). Activities of α‐amylase, lipase, trypsin, Na⁺, K⁺‐ATPase, alkaline phosphatase and gamma‐glutamyl transpeptidase in intestine were all affected by the dietary thiamin (P < 0.05). In conclusion, thiamin could improve growth and intestinal enzyme activities of juvenile Jian carp. The dietary thiamin requirement of juvenile Jian carp (8.0–60.2 g) based on PWG was 1.02 mg kg^?1 diet.     

Comparison of disease progress curves for yam anthracnose (Colletotrichum gloeosporioides)

The incidence and severity of foliar anthracnose, caused by Colletotrichum gloeosporioides , were recorded for seven fields of yams ( Dioscorea alata var. White Lisbon) in Barbados over a complete growing season. A scoring system was developed to estimate severity, placing the emphasis on the earliest, least visible stages of the disease. Disease progress curves were described in terms of the logistic model and the effects of site and leaf age on the parameters as well as on disease at tuber bulking were analysed. Differences between sites indicated that disease was more severe in areas of higher rainfall. In severe epidemics, disease predominated on young leaves. Fungicides applied sufficiently early can delay the onset of disease and slow the rate of disease progress.     

The induction of 1,3-β-glucanases and other enzymes in groundnut leaves infected with Cercospora arachidicola

Infection of groundnut leaves with the early leaf spot pathogen Cercospora arachidicola leads to a marked increase in extracellular 1,3-β-glucanase activity, limited to the infected tissue. Three isoforms of low molecular weight and extreme pI values, typical of pathogenesis-related proteins, were induced. These β-glucanases, when acting together, were capable of degrading the pathogen cell wall in vitro. Glucanases from homogenates of infected leaf tissue were partially purified by ion-exchange chromatography to give enzymes with molecular weights of 35, 32 and 20 kDa and pI values of 3·8, 3·6 and > 9, respectively. They were electrophoretically identical to the β-glucanases found in the intercellular washing fluid. Treatment of groundnut plants with 200 μM mercuric chloride induced the accumulation of identical extracellular β-glucanases. During the course of the infection an increase in peroxidase activity was also observed, but chitinase activity remained more or less constant.     

Assessing resistance to Crinipellis perniciosa using cocoa callus

Callus cultures from cocoa clones reported to have different susceptibilities to Brazilian isolates of Crinipellis perniciosa , causal fungus of witches' broom disease, including Catongo (susceptible). Scavina-6, CAB 64 and CAB 67 (resistant) were inoculated with basidiospores of C. perniciosa and the subsequent development of mycelia was assessed. Generally, on Catongo callus, mycelium similar to that found in green brooms (parasitic type), developed abundantly but on callus from Scavina-6 and the two CAB clones, mycelial growth was relatively poor and tended to be similar to that in dry brooms and on agar media (saprotrophic type). There was often considerable variation in fungal development on individual callus cultures from the same clone, but there was an overall consistency between successive experiments in ranking the callus material in terms of fungal development. However, growth of isolates from Colombia and Trinidad was similar on callus from Scavina-6 which in the field is known to reac differentially to these two isolates. Experiments with isolates from Colombia indicated that inoculum from basidiocarps produced on brooms which were kept for long periods in cabinets to induce fruiting had a decreased ability to colonize callus.