Persistent thrombin generation in humans during specific thrombin inhibition with hirudin

BACKGROUND: The degree to which antithrombotic drugs suppress thrombin generation is unknown. Because hirudin, unlike antithrombin III, binds intravascular thrombin rapidly and selectively to yield a circulating inactive complex of 3- to 4-hour half-life, we used intravenous hirudin in humans to investigate the course of thrombin generation during and early after anticoagulation with this potent, direct antithrombin. METHODS AND RESULTS: Intravascular thrombin was measured with an ELISA for the thrombin-hirudin complex formed during and for 18 hours after stopping a 6-hour infusion of hirudin at 0.1, 0.2, and 0.3 mg.kg-1.h-1 in three groups of six patients each. With free hirudin in 20- to 10,000-fold molar excess of thrombin and peak activated partial thromboplastin times of 2.3 to 3.0 times baseline, mean plasma thrombin-hirudin complex increased from 794 +/- 85 pg/mL (mean +/- SEM) 15 minutes after the start of the infusion to 1617 +/- 151 pg/mL at 6 hours of infusion to 2667 +/- 654 pg/mL at 24 hours. During the 24-hour observation period, plasma concentration of fragment 1.2 (the peptide released during conversion of prothrombin to thrombin) never fell below baseline but rather increased transiently during the hirudin infusion. Plasma concentrations of thrombin-antithrombin III complex (in ng/mL) decreased from 4.34 +/- 0.40 at baseline to 1.64 +/- 0.13 at 6 hours (P < .001) and gradually increased after stopping the infusion to 5.7 +/- 0.87 at 24 hours (nonsignificant compared with baseline). CONCLUSIONS: Measurement of thrombin-hirudin complex may be used as a marker of thrombin generation in humans. Persistent accumulation of thrombin-hirudin complex and generation of fragment 1.2 during and after completion of potent anticoagulation with hirudin suggest thrombin generation is not blocked by high-affinity thrombin inhibition. The persistent formation of thrombin during declining plasma levels of hirudin may contribute to the pathogenesis of rethrombosis early after antithrombin therapy or during inadequate anticoagulation.     

A Maternal High Fat Diet Has Long‐Lasting Effects on Skeletal Muscle Lipid and PLIN Protein Content in Rat Offspring at Young Adulthood

A maternal high fat diet (HFD) can have adverse effects on skeletal muscle development. Skeletal muscle PLIN proteins (PLIN2, 3 and 5) are thought to play critical roles in lipid metabolism, however effects of HFD on PLIN and lipases (HSL, ATGL, CGI‐58) in mothers as well as their offspring have yet to be investigated. The primary objective of this study was to determine whether maternal HFD would influence skeletal muscle lipase and PLIN protein content in offspring at weaning (19d) and young adulthood (3mo). Female rats (28d old, n = 9/group) were fed control (CON, AIN93G, 7 % soybean oil) or HFD (AIN93G, 20 % lard) for 10 weeks prior to mating and throughout pregnancy and lactation. All offspring were weaned to CON [n = 18/group, 1 female and 1 male pup per litter were studied at weaning (19d) and 3mo of age]. There was no effect of sex for the main outcomes measured in plantaris, therefore male and female data was combined. Maternal HFD resulted in higher triacylglycerol content in pups at 3mo (p < 0.05), as well as in the dams (p = 0.015). Maternal HFD resulted in higher PLIN5 content in pups at weaning and 3mo (p = 0.05). PLIN2 and PLIN5 content decreased at 3mo versus weaning (p < 0.001). HFD dams had a higher PLIN3 content (p = 0.016). Diet had no effect on ATGL, CGI‐58, or HSL content. In conclusion, exposure to a maternal HFD resulted in higher skeletal muscle lipid and PLIN5 content in plantaris of offspring through to young adulthood.     

The invasion of a large lake by the Eurasian genotype of common reed: The influence of roads and residential construction

The Eurasian genotype of common reed (Phragmites australis) is one of the most aggressive plant invading North American wetlands. There is, however, little published evidence on establishment patterns of populations along lakes of the St. Lawrence River–Great Lakes watershed. We tested the hypothesis that the recent invasion of Great Lake Saint-François (Québec, Canada) by common reed was facilitated by a dense road system and by an intense residence construction activity along lakeshores. A total of 345 and 2914 reed stands were mapped along lakeshores, and along the road system of the study area, respectively. The probability of finding a reed stand on a lakeshore increases with the proximity of the lake's outlet, and of a paved road, but decreases with the proximity of a residence built since 1990. It is likely that common reed first spread along the road system, and that wind dispersal of seeds then favored the establishment of populations on lakeshores. Our model does not support the hypothesis that residential construction facilitated the establishment of reed stands, probably because the recent residential construction boom occurred essentially in the southern part of the lake, where the number of roadside reed populations is much lower than in the northern part (lower seed rain). The invasion of Great Lake Saint-François shows that the spread of the plant is not restricted to major river or road systems. Large or small lakes, if submitted to intense diaspore pressure, can also be at risk.     

Probiotics: An alternative strategy for combating salmonellosis: Immune mechanisms involved

Salmonella produces infections of different nature and severity depending of many factors including the Salmonella serovar involved, strain virulence, infective dose, host animal species, age and immune status of the host. The treatments against Salmonella infections rely on supportive and antibiotic therapy to eliminate the pathogen, but the development of resistance by Salmonella to the antimicrobials most commonly used limits its efficacy. Other disadvantages of antibiotic treatments are that they can lead to acute diarrhea (antimicrobials normally induce an imbalance of intestinal bacterial flora) and may produce chronic toxicity. Considering this undesired consequences of antibiotics and because at the present there are no effective oral vaccines which protect against salmonellosis, scientists have been searching for alternative methods to control enteric infections. In the present review, probiotics are proposed as an attractive possibility to attend this concern. Probiotic are live microorganisms, which when administered in adequate amounts confer a health benefit on the host. In vitro and in vivo studies showed the effectiveness of probiotic administration in the prevention or in the treatment against Salmonella infection. There are several mechanisms by which probiotic strains might exert their effects. They include non immune mechanisms (stabilization of the gut mucosal barrier, competition for adhesion, secretion of antimicrobial substances, etc.) and the modulation of the mucosal and systemic immune responses. These mechanisms are species and/or strain specific. There are also evidences that in some cases, a mix of probiotic strains can be more useful than each strain alone against this infection. In addition, the presence of one or more probiotic strains in a fermented product can improve the beneficial properties of the probiotic strains involved. It was also reviewed the security of probiotics administration after Salmonella infection in healthy host and in immunosuppressed or babies hosts. Although, the major part of the researches were performed in animal models through in vivo assays or by in vitro studies using human cell lines, some studies carried out in humans to verify the probiotic effects were also addressed in the present review. Nevertheless, is of critical importance to perform more clinical trials in humans to validate the results obtained with each specific probiotic strain or probiotic product.     

Nutritional,functional, thermal and structural characteristics of <Emphasis Type="Italic">Citrullus lanatus</Emphasis> and <Emphasis Type="Italic">Limonia acidissima</Emphasis> seed flours

Seeds from fruits such as Citrullus (C.) lanatus (watermelon) and Limonia (L.) acidissima (wood apple) are not commonly utilized but could be suitable in numerous food formulations. It was shown that the protein content of defatted seed flours was 71.38 and 49.51 % and that these contained considerable amounts of minerals such as Na, Mn, Mg, K, Cu, Fe and Zn. The defatted L. acidissima seed flour was superior to C. lanatus in essential amino acids. The flours obtained from both seeds were also evaluated for functional properties and characterized by X-ray diffraction, differential scanning calorimeter and scanning electron microscope (SEM). Amorphous nature was observed in defatted C. lanatus and L. acidissima flours due to the low percentage of degree of crystallinity. Spherical morphologies were observed through SEM. The exothermic peak was recorded in defatted C. lanatus and L. acidissima flour.     

Effect of health status upon arrival at a single milk-fed veal facility on breakeven purchase price of calves

Male calves are purchased from the dairy industry in North America to produce red meat. The price paid for male calves varies widely, and it is unclear which variables influence the price paid for each calf. The objective of this study was to assess how the health traits of calves at the time of arrival and demographic variables affect the breakeven purchase price of a male calf entering the veal industry. A financial model was constructed using the prevalence of health abnormalities, weight at arrival, source of the calf, number of days in the barn, base carcass price, days to mortality, feed costs, season at arrival, interest rate, housing location, carcass dressing percentage, and costs associated with housing, labor, utilities, trucking, and health to calculate the breakeven purchase price and an estimate of profit. Sensitivity analysis was conducted using health variables measured at arrival and demographic variables, including season at arrival and housing location, to identify the factors with the largest impact on the predicted average daily gain, early and late mortality risk, breakeven purchase price, and profit. At the baseline inputs, the average calculated profit was ?$5.36 per calf and it was most sensitive to the location of housing where calves were fed and the body weight of the arriving calf. The mortality risk in the first 21 d after arrival (early) was calculated to be 2.2%, whereas the risk of mortality after 21 d (late) was 3.7%. The risks of early and late mortality were most sensitive to the level of dehydration measured at arrival and the season at arrival for the purchased calves, respectively. The calculated average daily gain was 1.12 kg/d and it was most sensitive to housing location. The breakeven purchase price was calculated to be $242.49 per calf, which was most sensitive to the housing location where the calves were fed. The results of this analysis demonstrate that veal producers need to consider many variables before purchasing calves. In addition to overall market conditions, veal producers should factor health characteristics and the expected performance of the calves they purchase into what they are willing to pay for them.     

Prepartum nutritional strategy affects reproductive performance in dairy cows

Negative energy balance during early postpartum is associated with reduced reproductive performance in dairy cows. A pooled statistical analysis of 7 studies completed in our group from 1993 to 2010 was conducted to investigate the association between prepartum energy feeding regimen and reproductive performance. The interval from calving to pregnancy (days to pregnancy, DTP) was the dependent variable to assess reproductive performance. Individual data for 408 cows (354 multiparous and 54 primiparous) were included in the analysis. The net energy for lactation (NE_L) intake was determined from each cow’s average dry matter intake and calculated dietary NE_L density. Treatments applied prepartum were classified as either controlled-energy (CE; limited NE_L intake to ≤100% of requirement) or high-energy (HE; cows were allowed to consume >100%) diets fed during the far-off (FO) or close-up (CU) dry periods. Cow was the experimental unit. The Cox proportional hazard model revealed that days to pregnancy was shorter for CE (median = 157 d) than HE (median = 167 d) diets during the CU period [hazard ratio (HR) = 0.70]. Cows fed HE diets during the last 4 wk prepartum lost more body condition score in the first 6 wk postpartum than those fed CE diets (−0.43 and −0.30, respectively). Cows fed CE diets during the FO period had lower nonesterified fatty acids concentrations in wk 1, 2, and 3 of lactation than cows fed HE diets. Higher nonesterified fatty acids concentration in wk 1 postpartum was associated with a greater probability of disease (n = 251; odds ratio = 1.18). Cows on the CE regimen during the FO period had greater plasma glucose concentrations during wk 1 and 3 after calving than cows fed the HE regimen. Higher plasma glucose (HG) concentration compared with lower glucose (LG) in wk 3 (HG: n = 154; LG: n = 206) and wk 4 (HG: n = 71; LG: n = 254) after calving was associated with shorter days to pregnancy (wk 3: median = 151 and 171 d for HG and LG, respectively, and HR = 1.3; wk 4: median = 148 and 167 d, respectively, and HR = 1.4). In the first 2 wk after calving, cows that received HE diets in the FO period had higher concentrations of total lipids and triglyceride and greater ratio of triglyceride to glycogen in liver than cows fed CE diets. In conclusion, cows fed CE diets during the CU period had a shorter interval between parturition and conception, which may be explained by increased NE_L intake during the first 4 wk postpartum and lower incidence of peripartal diseases. Lower body condition score loss during the first 6 wk postpartum and slightly higher glucose concentration at wk 3 likely contributed to improved reproductive performance.     

Associations between management practices and within-pen prevalence of calf diarrhea and respiratory disease on dairy farms using automated milk feeders

Data on management practices used with automated milk feeders (AMF) are needed to identify factors associated with calf health in these systems. The objectives of this observational, longitudinal, cross-sectional study were to estimate the prevalence of calf diarrhea (CD) and bovine respiratory disease (BRD), and to identify factors associated with prevalence of these diseases at the pen level on dairy farms feeding milk to group-housed calves with AMF. Seventeen dairy farms with AMF in Ontario, Canada, were visited 4 times, seasonally, over 1 yr. The clinical health of all calves (n = 1,488) in pens (n = 35) with AMF was scored to identify the number of calves with CD and BRD. Data on calf, feeder, and pen management practices were analyzed using generalized linear mixed regression models for each disease. Overall calf-level prevalence of CD and BRD were 23 and 17%, respectively. Median (interquartile range, IQR) within-pen prevalence of CD and BRD were 17% (7 to 37%) and 11% (0 to 28%), respectively. Median age (IQR) for diarrheic calves was 25 d (16 to 42 d), and for calves with BRD was 43 d (29 to 60 d). Factors associated with lower within-pen prevalence of CD were the administration of vitamin E and selenium at birth [odds ratio (OR) = 0.56; 95% confidence interval (CI): 0.32 to 0.99], feeding of probiotics (OR = 0.44, 95% CI: 0.22 to 0.93), and adding fresh bedding every 2 to 3 d (OR = 0.43; 95% CI: 0.24 to 0.76) compared with every 7 or more days. In contrast, sharing air with older cattle (>9 mo old) was associated with increased within-pen prevalence of CD (OR = 4.54, 95% CI: 1.88 to 10.52). Additionally, total bacteria counts ≥100,000 cfu/mL in milk samples taken from the AMF mixing jar were associated with increased within-pen prevalence of CD during the summer visit (OR = 3.34; 95% CI: 1.31 to 8.54). Increased total solids in milk or milk replacer (OR = 0.48, 95% CI: 0.27 to 0.85) and feeding whole milk versus milk replacer (OR = 0.29, 95% CI: 0.11 to 0.75) were associated with lower within-pen prevalence of BRD. Factors associated with greater within-pen prevalence of BRD were sharing air with weaned cattle up to 8 mo old (OR = 3.21, 95% CI: 1.26 to 8.16), and greater depth of the wet bedding pack. The use of maternity pens for reasons other than just calving was associated with increased prevalence of both CD and BRD (OR = 1.85, 95% CI: 1.03 to 3.33; OR = 2.61, 95% CI: 1.21 to 5.58, respectively). These results suggest that isolation from older animals and frequent cleaning of the feeder and pen may help to reduce disease prevalence in group-housed calves fed with an AMF.     

Synthesis and biological properties of novel pyridinioalkanoyl thiolesters (PATE) as anti-HIV-1 agents that target the viral nucleocapsid protein zinc fingers

Nucleocapsid p7 protein (NCp7) zinc finger domains of the human immunodeficiency virus type 1 (HIV-1) are being developed as antiviral targets due to their key roles in viral replication and their mutationally nonpermissive nature. On the basis of our experience with symmetrical disulfide benzamides (DIBAs; Rice et al. Science 1995, 270, 1194-1197), we synthesized and evaluated variants of these dimers, including sets of 4,4'- and 3,3'-disubstituted diphenyl sulfones and their monomeric benzisothiazolone derivatives (BITA). BITAs generally exhibited diminished antiviral potency when compared to their disulfide precursors. Novel, monomeric structures were created by linking haloalkanoyl groups to the benzamide ring through -NH-C(=O)- (amide) or -S-C(=O)- (thiolester) bridges. Amide-linked compounds generally lacked antiviral activity, while haloalkanoyl thiolesters and non-halogen-bearing analogues frequently exhibited acceptable antiviral potency, thus establishing thiolester benzamides per se as a new anti-HIV chemotype. Pyridinioalkanoyl thiolesters (PATEs) exhibited superior anti-HIV-1 activity with minimal cellular toxicity and appreciable water solubility. PATEs were shown to preferentially target the NCp7 Zn finger when tested against other molecular targets, thus identifying thiolester benzamides, and PATEs in particular, as novel NCp7 Zn finger inhibitors for in vivo studies.     

Diabetic-like retinopathy: early and late intervention therapies in galactose-fed rats

PURPOSE: To determine whether the diabetic-like thickening of retinal capillary basement membrane (RCBM) that develops in the galactose-fed rat model of diabetic ocular complications could be halted or ameliorated after 4 or 8 months of galactosemia by treatment with ARI-509, a potent new aldose reductase inhibitor (ARI), or by withdrawal of the galactose diet. METHODS: Weanling female Sprague-Dawley rats were randomized into eight groups and fed laboratory chow plus 50% starch, control group (CON); 50% D-galactose, galactose-fed group (GAL); 50% D-galactose with ARI-509 at 25 mg/kg or 10 mg/kg body wt per day, high-dose prevention group (HDP) and low-dose prevention group (LDP), respectively; 50% D-galactose for 4 or 8 months and then intervention by addition of ARI-509 (25 mg/kg body wt per day), 4-month intervention group (4IN) and 8-month intervention group (8IN), respectively; or 50% D-galactose for 4 or 8 months and then intervention by withdrawing galactose and replacing it with the 50% starch diet, 4-month galactose withdrawal group (4GW) and 8-month galactose withdrawal group (8GW), respectively. After 4, 8, 16, and 24 months of the experimental diets, the levels of carbohydrates in tissues and the extent of RCBM thickening in capillaries of the outer plexiform layer were determined in all groups. RESULTS: Retinal polyol was reduced by 95% in all ARI-treated groups and by 100% in the 4GW and 8GW groups after withdrawal of the galactose. The mean RCBM thickness increased rapidly in GAL rats, becoming almost two times greater (189 +/- 9.4 nm) than in CON rats (103 +/- 3.4 nm) by 24 months. Treatment with ARI-509 in high and low doses (HDP, LDP) initiated with the introduction of the galactose diet significantly prevented RCBM thickening at all time points (P < 0.05). In contrast, intervention by withdrawing galactose from the diet or by adding the high dose of ARI-509 had no significant effect (P < 0.05) on RCBM thickening until the 24-month time point (4IN, 166 +/- 10.3 nm; 8IN, 161 +/- 8.2 nm; 4GW, 136 +/- 5.1 nm; 8GW, 163 +/- 9.6 nm). CONCLUSIONS: Both early and late interventions decreased RCBM thickening compared with that in untreated GAL rats. The decreased thickening, however, was not evident until 16 to 20 months after the intervention. Because RCBM thickening is one of the earliest changes in diabetic and galactosemic retinopathy, the findings suggest that RCBM thickening and possibly subsequent retinal lesions are caused by early biochemical alterations induced by the galactose diet that are not readily reversed. The delayed response to therapy is consistent with that observed in the Diabetes Control and Complications Trial. The cumulative evidence indicates that intervention should begin as early after onset of diabetes as possible, and long follow-up periods should be used to evaluate efficacy.