Ovine sperm DNA oxidation quantification using an 8‐OHdG immunodetection assay

Our aim was to optimize 8‐hydroxy‐2′‐deoxyguanosine (8‐OHdG) immunodetection in order to detect DNA damage caused by oxidative stress that may not be detected by other DNA integrity analysis techniques, especially due to the high compaction of DNA in ruminants. Semen samples from 6 rams were cryopreserved. After thawing, samples were subjected to the DNA oxidation quantification using an 8‐OHdG immunodetection assay by flow cytometry. We have evaluated two different incubation times (30 min vs. overnight) at 4°C of the primary antibody (monoclonal anti‐8‐OHdG antibody). We have also compared the results of this technique with the sperm chromatin structure assay (SCSA^®). The analysis revealed that there were no significant differences (p > .05) between different incubation times. However, overnight incubation seems to cause more non‐specific binding of the secondary antibody. Significant differences (p < .05) between subjects and oxidation controls (8 M H₂O₂/800 μM FeSO₄?7H₂O) were evident. We can conclude that the 8‐OHdG immunodetection assay for DNA oxidation quantification of ram sperm can be performed subjecting sperm samples to a very high oxidative treatment.     

Yolk coelomitis in Fiji Island banded iguanas (Brachylophus fasciatus)

Yolk coelomitis is a major cause of death in captive sexually mature female Fiji Island banded iguanas (Brachylophus fasciatus) maintained by the Zoological Society of San Diego. The medical records, breeding histories, and pathology archival materials from this group were reviewed to study this health problem. From 1987 through 2004, deaths of nine of 21 adult females were due to yolk coelomitis. Most iguanas had a history of reproduction-related problems, which included reproductive failure, episodes of lethargy associated with ovarian activity, folliculostasis, ovostasis, and behavioral abnormalities. Most affected iguanas either were found dead or presented moribund and subsequently died or were euthanized. Clinical signs were nonspecific and included lethargy, cutaneous discoloration, and coelomic effusion. Yolk leakage in most cases was associated with the presence of large vitellogenic follicles undergoing atresia and resulted in coelomitis characterized by florid mesothelial proliferation.     

Responses of lettuce to salinity. I. Effects of NaCl and Na2SO4 on growth

Two lettuce cultivars (Lactuca sativa L., cv. Calmar and cv. Climax) were selected to compare their tolerance to salt stress. The plants were grown in a hydroponic system using a 0.5 modified Hoagland solution. Treatments of 0, 40, 80, and 120 mol m^–3 NaCl or 0 and 20 mol m^–3 Na₂SO₄ were started when the second leaf above the cotyledons appeared. The plants were harvested 20 days later. Climax showed a greater tolerance to salinity at the 40 mol m^–3 NaCl concentration; the % decrease in both shoot and root fresh weight was significantly less than Calmar. No differences between the cultivars were found in the Na₂SO₄ experiments. Differences in root Cl^– content at the 40 mol m^–3 concentration corresponded to an enhanced water content of the roots. A mechanism for the observed differences in salt tolerance between the two cultivars is discussed.     

Leakage of a flavin‐like compound from lettuce (Lactuca sativa L.) under osmotic stress

Abstract

An unusual response vas noted in the salinized nutrient solutions of lettuce (Lactuca sativa L.) in that they became clear yellow in color. This condition also developed in experiments using polyethylene glycol in place of salt, suggesting that the response vas due to the osmotic stress imposed upon the roots. Photometric and fluorometric determinations indicated that the color could be attributed to a flavin‐like compound or compounds released by the roots.     

The advantages and limitations of trait analysis with GWAS: a review

Over the last 10 years, high-density SNP arrays and DNA re-sequencing have illuminated the majority of the genotypic space for a number of organisms, including humans, maize, rice and Arabidopsis. For any researcher willing to define and score a phenotype across many individuals, Genome Wide Association Studies (GWAS) present a powerful tool to reconnect this trait back to its underlying genetics. In this review we discuss the biological and statistical considerations that underpin a successful analysis or otherwise. The relevance of biological factors including effect size, sample size, genetic heterogeneity, genomic confounding, linkage disequilibrium and spurious association, and statistical tools to account for these are presented. GWAS can offer a valuable first insight into trait architecture or candidate loci for subsequent validation.     

Apoptotic effects of tamoxifen on leukocytes from horse peripheral blood and bronchoalveolar lavage fluid

A reduction in inflammatory cell apoptosis is an important concept in the maintenance of inflammation and a potential target for the resolution of inflammation in many inflammatory diseases. Dysregulation of apoptosis has been implicated in a range of diseases, including tumors, neurodegenerative disorders and autoimmunity, and may also be implicated in allergic asthma. In horses, recurrent airway obstruction (RAO) is an asthma-like condition that is characterized increased survival neutrophil bronchial. Tamoxifen is a synthetic, non-steroidal, anti-estrogen agent that is widely used for treating all stages of breast cancer and has been approved for the prevention of breast cancer in high-risk women. The observed efficacy of tamoxifen has been attributed to both growth arrest and the induction of apoptosis. Therefore, the aim of our study was to evaluate the ability of tamoxifen to induce apoptosis in vitro in granulocytic cells from peripheral blood and in mononuclear cells from bronchoalveolar lavage fluid (BALF) in horses. Flow cytometry using commercial AnnexinV-FITC and propidium iodide was used to quantify early and late apoptotic leukocytes, respectively. The results showed a significant increase in early apoptosis in peripheral blood and bronchial granulocytic cells treated with tamoxifen. The rate of early apoptosis of mononuclear cells from blood and BALF when incubated with tamoxifen was significantly lower compared with granulocytic cells. We did not observe a direct effect of tamoxifen on late apoptosis in any of the in vitro assays in the cell types used here. These results indicate that the apoptotic mechanisms under these experimental conditions would affect only blood and BALF granulocytic cells, particularly in early apoptosis. Finally, further in vitro and in vivo studies are needed to better understand apoptotic mechanisms because tamoxifen could be used to treat chronic, inflammatory pathologies associated with granulocytes and allergic diseases, such as asthma or equine RAO.     

Prediction of long‐term sustainability of constructed urban soil: impact of high amounts of organic matter on soil physical properties and water transfer

Sustainability of urban soils lies in their ability to facilitate water and air permeabilities. Exogenous organic matter has been shown to have a positive impact on these properties. Under urban conditions, a large one‐time input of an organic amendment was made to the reconstituted soil. Two organic materials, green‐waste compost (gw) or cocompost from sewage sludge and wood chips (sw), were mixed with sandy loam soil (40% v/v) and placed in 600‐L containers. Containers received a 29‐cm thick layer of sandy loam soil–organic matter mix over a 28‐cm thick layer without organic amendment. Volumetric water content, dry bulk density, hydraulic conductivity at saturation and water retention were measured over 5 yrs in the soils and values for the mixes and a control compared. After this time, dry bulk density was greater (1.54 g/cm³) in control than in gw or sw soils (1.31 and 1.11 g/cm³, respectively), whereas hydraulic conductivity at saturation was smaller (4 × 10^?7 m/s) than in gw (3.4 × 10^?6) or sw (3.7 × 10^?6 m/s). HYDRUS 1D water balance model indicated that below 27 cm depth in the control after 5 yrs, there was a high degree of anoxia, lasting >200 days per year, compared with <40 days in gw and sw. Amplification of the risk of anoxia below 27 cm depth after 10 yrs was 323, 151 and 100 days in the control, gw and sw, respectively. Organic matter amendment could support sustainable urban soils for ten years after soil reconstitution.