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     

Molecular characterisation of Theileria equi and risk factors associated with the occurrence of theileriosis in horses of Punjab (Pakistan)

Theileria equi (T. equi) is an obligate intra- and extra-erythrocytic parasite that causes equine theileriosis (ET) in equids. Equine theileriosis is considered a notifiable disease of global significance, a major constraint to the international movement of horses, and endemic in many countries. This disease may be difficult to diagnose, as it can produce variable and nonspecific clinical signs. A cross-sectional study was designed for the molecular characterisation of T. equi and to investigate the associated risk factors of ET accompanied by its consequences on haematological and sero-biochemical parameters. A convenience sampling of 500 blood samples were collected from ET suspect horses from January to December 2017. PCR was performed on all blood samples targeting the 18S rRNA gene of T. equi followed by sequencing; 9% animals tested positive with confirmed sequences. The isolates of this study showed high homology with Cuban, Russian and Brazilian isolates of T. equi (accession numbers KY111762.2 , MG551915.1 and KY952237.1 , respectively). Based on multivariate analysis, the principal risk factors consisted of absence of dogs on the premises and presence of tick infestation. The haemato-biochemical parameters showed a decrease in granulocytes and erythrocytes, and an increase in lymphocytes, monocytes, mean corpuscular volume, mean corpuscular haemoglobin, mean platelet volume, glucose, phosphorus and aspartate aminotransferase in positive horses. This is the first study which identified ET in Punjab (Pakistan) using molecular techniques and risk factors together with the haemato-biochemical variations in horses.     

Challenges for the sustainable use of water and land resources under a changing climate and increasing salinization in the Jizzakh irrigation zone of Uzbekistan

Jizzakh Province in Uzbekistan is one of the largest irrigated areas in Central Asia without natural drainage.In combination with aridity,climate change and extensive irrigation practices,this has led to the widespread salinization of agricultural land.The aim of this study was to identify opportunities to improve the reclamation status of the irrigated area and how best to effectively use the water resources in Jizzakh Province based on investigations conducted between 1995 and 2016.A database of field measurements of groundwater levels,mineralization and soil salinity conducted by the provincial Hydro-Geological Reclamation Expeditions was used in the study.The total groundwater mineralization was determined using a portable electric conductometer(Progress 1T)and the chloride concentration was determined using the Mohr method.The soil salinity analyses were conducted by applying two different methods:(1)the extraction and assessment of the soluble salt content,and(2)using an SM-138 conductivity sensor applied to a 1:1 mixture of soil sample and water.The analyses of the monitoring results and the salt balance in the"irrigation water–soil–drainage water"system clearly demonstrated that the condition of the irrigated land in the province was not significantly improved.Under these conditions,the stability of crop yields is achieved mainly through the use of large volumes of fertilizer.However,excess amounts of mineral fertilizers can also cause the salinization of soils.The average groundwater salinization value in most of the irrigated land(75.3%)fluctuated between 1.1 and 5.0 g/L,while the values were less than 1.0 g/L in 13.1%of the land and in the range of 5.1–10.0 g/L in 10.5%of the land.During the period of 1995–2016 the salinization level of the irrigated land in Jizzakh Province increased slightly and the area could be divided into the following classes:no salinity(17.7%of the total area),low salinity(51.3%),moderate salinity(29.0%),and high salinity(2.0%).Detailed studies of the salt balance in irrigated land,the impact of climate change,increased fertilizer use,and repeated remediation leaching on the groundwater level and mineralization should be conducted in the future,due to the possibility of accelerated salinization,fertility decline,and reduced yields of agricultural crops.     

Soil amendments and seed priming influence nutrients uptake,soil properties,yield and yield components of wheat (Triticum aestivum L.) in alkali soils

The effects of soil amendments [i.e., control, gypsum, farmyard manure (FYM), and gypsum?+?FYM] and seed priming (i.e., unprimed, seed soaked in water for 10?hr prior to sowing, and seed soaked in 0.4% gypsum solution for 10?hr prior to sowing) were assessed on growth and yield of wheat (Triticum aestivum L.) crop in alkali soil in northwestern Pakistan. A split plot design was used, keeping priming methods in main plots and soil amendments in sub-plots. The results showed that the effects of soil amendments and seed priming on grain yield, straw yield, harvest index and number of spikes were significant but their interactive effect was non-significant. The highest crop yields and yield index were obtained with gypsum?+?FYM amendments, and seed priming with gypsum solution. The effect on seed emergence, plant height and number of grains per spike was, however, not significant. Grain yield increased by 104% in gypsum?+?FYM treatment over control and by 16.8% with seed primed in water, followed by 8.5% with priming in gypsum solution, as compared to non-priming. The weight of 1000 grains was significantly increased by 35% in gypsum?+?FYM treatment and by 15.8% in gypsum priming. The phosphorus (P) and potassium (K) content increased with soil amendments. Soil pH and gypsum requirement reduced significantly with soil amendments. The blend of gypsum and FYM has improved the properties of salt-affected soil and enhanced fertility for optimum production of wheat in addition to the beneficial effect of seed priming in gypsum solution on crop yield. Using these amendments could be ameliorative in removing the adverse effect of the salt-affected soils, rendering the soil a good medium for plant growth.     

Chemical Constituents of Lagenaria siceraria Mesocarp and Its Xanthine Oxidase and Alpha-Amylase Inhibitory Activities

The present study was conducted to identify natural products in the bioactive fractions of Lagenaria siceraria mesocarp. Column chromatography and gas chromatography-mass spectrometry (GC-MS) was used for separation and identification of phytochemicals. Methanolic extract and its fractions displayed alpha-amylase and xanthine oxidase inhibitory activities comparable to standard drugs. GC-MS analysis of the fractions and column eluates revealed the presence of a variety of chemical compounds, including 4-(methoxymethyl)phenol, 2,2?-methylenebis{6(1,1-dimethylethyl)-4-ethyl}phenol, methyl 2-hydroxy-3-phenylpropanoate, oxacyclododecane-2,8-dione, N-(2-(4-hydroxyphenyl)ethyl] acetamide, 2,3-Epoxycarane, 4-(propan-2-yl)benzaldehyde, octadec-1-ene, 4a-methyl-1-methylidene-1,2,3,4,4a,9,10,10a-octahydrophenanthrene, 2,6-di(propan-2-yl)naphthalene, methyl 3-(4-hydroxy-3,5-dimethylphenyl)propanoate, 4,8a-dimethyl-6-(prop-1-en-2-yl)-3,5,6,7,8,8a-hexahydronaphthalen-2(1H)-one 1,3-dihydro-2-benzofuran, methyl N-hydroxybenzenecarboximidoate, linoleic acid, and palmitic acid. Many of these compounds are being reported for the first time from this plant, the presence of which may explain, at least partly, its medicinal efficacy. The fruit possesses significant anti-enzymatic properties against alpha-amylase and xanthine oxidase, and the most active compounds of mesocarp of the fruit appear in ethyl acetate fraction of its methanolic extract.     

Investigation on hydrostatic resistance and thermal performance of layered waterproof breathable fabrics

Waterproof breathable layered fabrics allow water vapor passing through, but resist liquid water to pass. This ability of the fabrics to protect rain and snow water while allowing sweat vapor to evaporate from inside to outside atmosphere, leads them to be used as outdoor sportswear or protective clothing. The big challenge of enhanced hydrostatic resistance of these fabrics with proper breathability and thermal comfort has widened the research scope. This study presents an experimental investigation on hydrostatic resistance and thermal behavior of layered waterproof breathable fabrics. Six different types of hydrophobic and hydrophilic membrane laminated layered fabrics were evaluated by varying different fabric parameters in the experiment. Hydrostatic resistance and water vapor permeability of the laminated fabrics were measured by SDL ATLAS Hydrostatic Head Tester and PERMETEST respectively. Thermal properties were evaluated by ALAMBETA instrument. Moreover, FX-3300 air permeability tester was used to measure air permeability which represents the porosity of the fabrics and computer based See System software was used for water contact angle measurement on the outer fabric surface in order to determine the hydrophobic and hydrophilic properties. This experiment clearly discusses the influence of different fabric characteristics and parameters on hydrostatic resistance and thermal properties of the breathable laminated fabrics. The results show that fabric material composition, density, thickness, and hydrophobic and hydrophilic membranes have significant effects on hydrostatic resistance, breathability and thermal properties of different laminated fabrics.     

Involvement of CD28/CTLA4-B7 costimulatory pathway in the development of lymphadenopathy and splenomegaly in MRL/lpr mice

MRL/lpr mouse is an established animal model which develops autoimmune diseases including glomerulonephritis, sialoadenitis, hepatitis and inflammatory lung disease. Additionally, it has been reported that lpr strains uniquely accumulate CD3+ CD4- CD8- B220+ (double negative, DN) T cells in lymphoid organs leading to lymphadenopathy and splenomegaly. To investigate the role of CD28/CTLA4-B7 pathway in the development of lymphadenopathy and splenomegaly, MRL/lpr mice were treated with soluble form of CTLA4 molecules, CTLA4IgG, which efficiently blocks this pathway. It was demonstrated that (i) the development of DN T cells was independent of the CD28/CTLA4-B7 pathway, (ii) the CD28/CTLA4-B7 pathway was required for the development of lymphadenopathy and splenomegaly, (iii) the CD28/CTLA4-B7 pathway was important for the accumulation of various cell populations in the lymph node and spleen, (iv) composition of the accumulating cell populations was not altered by CTLA4IgG treatment, and (v) activation of conventional T cells and IL-4 production from conventional T cells were the CD28/CTLA4-B7 pathway dependent. Thus, we concluded that the CD28/CTLA4-B7 pathway was required for the development of full-blown lymphadenopathy and splenomegaly in MRL/lpr mice.     

The Sepsis-Coagulant Axis: A Review

Activation of coagulation is a normal component of the acute inflammatory response. Inflammatory cytokines initiate coagulation events locally at sites of inflammation by converting endothelium from an antithrombotic surface to a prothrombotic surface; by stimulating tissue factor production, which activates both the extrinsic and intrinsic coagulation systems; and by stimulating production of platelet-activating factors. The fibrinolytic system is initially activated but is subsequently inhibited. This results in a marked imbalance in coagulation and fibrinolysis resulting in a net procoagulant state. When thrombin generation and platelet activation exceed the body's capacity to inactivate or remove these factors, disseminated intravascular coagulation (DIC) results. DIC directly contributes to multiple organ failure and death associated with sepsis. Presently available treatments (ie, heparin and aspirin) are relatively ineffective in treating DIC; however, newer, more potent drugs may soon be available for clinical use.     

Antibacterial activity and cytotoxicity of extractives from Ravenia spectabilis

A methanolic extract of Ravenia spectabilis, an isolated alkaloid, arborinine plus a fraction comprising arborinine and gamma-fagarine (VLC), showed mild to significant in vitro antibacterial activity. In a brine shrimp lethality bioassay, the extract and the fraction were found to exhibit moderate cytotoxicity having LC50 of 76.26 microg/ml and 14.98 microg/ml, respectively.