Mediation of poly(ADP-ribose) polymerase-1-dependent cell death by apoptosis-inducing factor

Poly(ADP-ribose) polymerase-1 (PARP-1) protects the genome by functioning in the DNA damage surveillance network. PARP-1 is also a mediator of cell death after ischemia-reperfusion injury, glutamate excitotoxicity, and various inflammatory processes. We show that PARP-1 activation is required for translocation of apoptosis-inducing factor (AIF) from the mitochondria to the nucleus and that AIF is necessary for PARP-1-dependent cell death. N-methyl-N'-nitro-N-nitrosoguanidine, H2O2, and N-methyl-d-aspartate induce AIF translocation and cell death, which is prevented by PARP inhibitors or genetic knockout of PARP-1, but is caspase independent. Microinjection of an antibody to AIF protects against PARP-1-dependent cytotoxicity. These data support a model in which PARP-1 activation signals AIF release from mitochondria, resulting in a caspase-independent pathway of programmed cell death.     

Plasma endotoxin concentration in horses: a methods study

Background: Accurate determination of plasma endotoxin concentration is critical for ex vivo and in vitro cellular and molecular studies of endotoxemia in horses. However, reports are conflicting with respect to anticoagulant, handling, and sample preparation.

Objective:

Objective: The purpose of this study was to determine the effect of blood sample fraction and handling time on measurement of endotoxin concentration in horses.

Methods:

Methods: Whole blood, anticoagulated with 3.8% (0.12 M) sodium citrate (9:1), was collected from 5 healthy horses. Whole blood (WB), platelet-rich plasma (PRP), and platelet-poor plasma (PPP) were spiked with endotoxin (2 EU/mL). Endotoxin-spiked WB samples were centrifuged immediately to generate PRP for measurement. Endotoxin concentration was subsequently measured by Limulus amebocyte assay at 0, 15, 30, 45, and 60 minutes. Assays were performed in triplicate and results were analyzed using Student's t -test, with significance set at P < .05.

Results:

Results: Mean endotoxin concentrations in 2 EU/mL-spiked WB were significantly different from those in PPP at all time points tested. Recovery of endotoxin in PRP generated from WB was significantly diminished after just 15 minutes.

Conclusion:

Conclusion: PRP generated from WB is significantly more reliable than PPP in determining endotoxin concentration ex vivo. Measurement of endotoxin in PRP generated from WB was significantly diminished after 15 min, identifying a time frame within which to process blood samples for endotoxin analysis.     

Cytokine responses to EHV-1 infection in immune and non-immune ponies

Protecting equids against equine herpesvirus-1 (EHV-1) infection remains an elusive goal. Repeated infection with EHV-1 leads to protective immunity against clinical respiratory disease, and a study was conducted to measure the regulatory cytokine response (IFN-gamma and IL-4) in repeatedly infected immune ponies compared to non-immune ponies. Two groups of four ponies were established. Group 1 ponies had previously been infected on two occasions, and most recently 7 months before this study. Group 2 ponies had no history no vaccination or challenge infection prior to this study. Both groups were subjected to an intranasal challenge infection with EHV-1, and blood samples were collected pre-infection, and at 7 and 21 days post-infection for preparation of PBMCs. At each time point, the in vitro responses of PBMCs to stimulation with EHV-1 were measured, including IFN-gamma and IL-4 mRNA production, and lymphoproliferation. Group 1 ponies showed no signs of clinical disease or viral shedding after challenge infection. Group 2 ponies experienced a biphasic pyrexia, mucopurulent nasal discharge, and nasal shedding of virus after infection. Group 1 ponies had an immune response characterized both before and subsequent to challenge infection by an IFN-gamma response to EHV-1 in the absence of an IL-4 response, and demonstrated increased EHV-1-specific lymphoproliferation post-infection. Group 2 ponies had limited cytokine or lymphoproliferative responses to EHV-1 pre-challenge, and demonstrated increases in both IFN-gamma and IL-4 responses post-challenge, but without any lymphoproliferative response. Protective immunity to EHV-1 infection was therefore characterized by a polarized IFN-gamma dependent immunoregulatory cytokine response.     

Structure–activity relationships of eugenol derivatives against Aedes aegypti (Diptera: Culicidae) larvae

BACKGROUND: Dengue fever is a severe public health problem for several countries. In order to find effective larvicides to aid control programs, the structure‐activity relationships of eugenol derivatives against Aedes aegypti (Diptera: Culicidae) larvae were evaluated. Additionally, the composition and larvicidal activity of Syzygium aromaticum essential oil was assessed. RESULTS: Four compounds representing 99.05% of S. aromaticum essential oil have been identified. The essential oil was active against Ae. aegypti larvae (LC₅₀ = 62.3 and 77.0 ppm, field‐collected and Rockefeller larvae respectively). The larvicidal activity of eugenol, the major compound of the essential oil, was further evaluated (LC₅₀ = 93.3 and 71.9 ppm, field‐collected and Rockefeller larvae respectively). The larvicidal activity and structure‐activity relationships of synthetic derivatives of eugenol were also assessed. The larvicidal activity of the derivatives varied between 62.3 and 1614.9 ppm. Oxidation of eugenol allylic bond to a primary alcohol and removal of the phenolic proton resulted in decreased potency. However, oxidation of the same double bond in 1‐benzoate‐2‐methoxy‐4‐(2‐propen‐1‐yl)‐phenol resulted in increased potency. CONCLUSION: Structural characteristics were identified that may contribute to the understanding of the larvicidal activity of phenylpropanoids. The present approach may help future work in the search for larvicidal compounds. Copyright © 2012 Society of Chemical Industry     

Congenital limb abnormality in a Thoroughbred foal in Trinidad

A congenital defect of the right hindlimb of a 36‐h‐old Thoroughbred foal as reported for the first time in Trinidad, West Indies. The foal was subjected to euthanasia and radiographs of both hindlimbs revealed that the bones distal to P1 of the right hindlimb only, were missing. Insertion of both the superficial and deep digital flexor tendons were found to be on the most proximal aspect of P1. Necropsy revealed no other congential defects.     

Variation in seed and seedling traits among natural populations of <Emphasis Type="Italic">Trigonobalanus doichangensis</Emphasis> (A. Camus) Forman (Fagaceae), a rare and endangered plant in southwest China

The extent of genetic variation in seed and seedling traits of Trigonobalanus doichangensis, a rare and endangered tree in southwest China, was evaluated at the population level. Seeds were collected from four natural populations in the south and southwest Yunnan province, China. Significant differences (P < 0.05) between populations were observed in seed length, seed weight and seedling morphological characters. The analyses revealed that none of the seed and seedling traits had significant correlations with the geo-climatic variables of the populations. Among seed and seedling traits, significant correlations (P < 0.05) were discovered among seed length, seed weight and height of 30-day-old seedlings. Significant correlation (r = 1.00, P < 0.01) occurred also between the height and root collar diameter of 4-year-old seedlings. With the exception of seed width and seed germination percentage, 51–96% of total variation in other seed and seedling related traits was attributed to population effect, as determined by the broad sense heritability. In conclusion, these observations on variation in seed and seedling traits should be a reference point, when considering seed collection of this species for ex situ conservation and species restoration.     

Conserving genomic variability in large mammals: Effect of population fluctuations and variance in male reproductive success on variability in Yellowstone bison

Loss of genetic variation through genetic drift can reduce population viability. However, relatively little is known about loss of variation caused by the combination of fluctuating population size and variance in reproductive success in age structured populations. We built an individual-based computer simulation model to examine how actual culling and hunting strategies influence the effective population size (N_e) and allelic diversity in Yellowstone bison over 200 years (～28 generations). The N_e for simulated populations ranged from 746 in stable populations of size 2000 up to 1165 in fluctuating populations whose census size fluctuates between 3000 and 3500 individuals. Simulations suggested that ～93% of allelic diversity, for loci with five alleles will be maintained over 200 years if the population census size remains well above ～2000 bison (and if variance in male reproductive success is high). However for loci with 20 alleles, only 83% of allelic diversity will be maintained over 200 years. Removal of only juveniles (calves and yearlings) resulted in longer generation intervals which led to higher maintenance of allelic diversity (96%) after 200 years compared to the culling of adults (94%) when the mean census size was 3250 (for loci with five alleles). These simulations suggest that fluctuations in population census size do not necessarily accelerate the loss of genetic variation, at least for the relatively large census size and growing populations such as in Yellowstone bison. They also suggest that the conservation of high allelic diversity (>95%) at loci with many alleles (e.g., ?5) will require maintenance of a populations size greater than approximately 3250 and removal of mainly or only juveniles.