DNA double strand breaks but not interstrand crosslinks prevent progress through meiosis in fully grown mouse oocytes

There is some interest in how mammalian oocytes respond to different types of DNA damage because of the increasing expectation of fertility preservation in women undergoing chemotherapy. Double strand breaks (DSBs) induced by ionizing radiation and agents such as neocarzinostatin (NCS), and interstrand crosslinks (ICLs) induced by alkylating agents such as mitomycin C (MMC), are toxic DNA lesions that need to be repaired for cell survival. Here we examined the effects of NCS and MMC treatment on oocytes collected from antral follicles in mice, because potentially such oocytes are readily collected from ovaries and do not need to be in vitro grown to achieve meiotic competency. We found that oocytes were sensitive to NCS, such that this ionizing radiation mimetic blocked meiosis I and caused fragmented DNA. In contrast, MMC had no impact on the completion of either meiosis I or II, even at extremely high doses. However, oocytes treated with MMC did show ?-H2AX foci and following their in vitro maturation and parthenogenetic activation the development of the subsequent embryos was severely compromised. Addition of MMC to 1-cell embryos caused a similarly poor level of development, demonstrating oocytes have eventual sensitivity to this ICL-inducing agent but this does not occur during their meiotic division. In oocytes, the association of Fanconi Anemia protein, FANCD2, with sites of ICL lesions was not apparent until entry into the embryonic cell cycle. In conclusion, meiotic maturation of oocytes is sensitive to DSBs but not ICLs. The ability of oocytes to tolerate severe ICL damage and yet complete meiosis, means that this type of DNA lesion goes unrepaired in oocytes but impacts on subsequent embryo quality

Allergenicity of Various Peanut Products as Determined by RAST Inhibition

Extracts of 19 different peanut products and peanut oil were tested for their allergenicity by the radioallergosorbent test inhibition assay using a crude peanut extract from raw peanuts as the standard for comparison. Seventeen of the extracts were able to competitively inhibit the binding of serum IgE from peanut-sensitive patients with the solid-phase raw peanut extract. Peanut oil and the extract from hydrolyzed peanut protein did not inhibit binding, which suggests that these products are not allergenic. The peanut hull flour extract showed a slight ability to inhibit binding, suggesting that this product contains minor amounts of the peanut allergen

Identification of Epigenetic Regulators of DUX4-fl for Targeted Therapy of Facioscapulohumeral Muscular Dystrophy

Facioscapulohumeral muscular dystrophy (FSHD) is caused by epigenetic de-repression of the disease locus, leading to pathogenic misexpression of the DUX4 gene in skeletal muscle. While the factors and pathways involved in normal repression of the FSHD locus in healthy cells have been well characterized, very little is known about those responsible for the aberrant activation of DUX4-fl in FSHD myocytes. Reasoning that DUX4-fl activators might represent useful targets for small molecule inhibition, we performed a highly targeted, candidate-based screen of epigenetic regulators in primary FSHD myocytes. We confirmed several of the strongest and most specific candidates (ASH1L, BRD2, KDM4C, and SMARCA5) in skeletal myocytes from two other unrelated FSHD1 patients, and we showed that knockdown led to reduced levels of DUX4-fl and DUX4-FL target genes, as well as altered chromatin at the D4Z4 locus. As a second mode of validation, targeting the CRISPR/dCas9-KRAB transcriptional repressor to the promoters of several candidates also led to reduced levels of DUX4-fl. Furthermore, these candidates can be repressed by different methods in skeletal myocytes without major effects on certain critical muscle genes. Our results demonstrate that expression of DUX4-fl is regulated by multiple epigenetic pathways, and they indicate viable, druggable candidates for therapeutic target development

Impact of varying analytical methodologies on grain particle size determination

Citation: Kalivoda, J. R., Jones, C. K., & Stark, C. R. (2017). Impact of varying analytical methodologies on grain particle size determination. Journal of Animal Science, 95(1), 113-119. doi:10.2527/jas2016.0966The determination of particle size is an important quality control measurement for feed manufacturers, nutritionists, and producers. The current approved method for determining the geometric mean diameter by weight (d(gw)) and geometric standard deviation (S-gw) of grains is standard ANSI/ASAE S319.4. This method controls many variables, including the suggested quantity of initial material and the type, number, and size of sieves. However, the method allows for variations in sieving time, sieve agitators, and the use of a dispersion agent. The objective of this experiment was to determine which method of particle size analysis best estimated the particle size of various cereal grain types. Eighteen samples of either corn, sorghum, or wheat were ground and analyzed using different variations of the approved method. Treatments were arranged in a 5 x 3 factorial arrangement with 5 sieving methods: 1) 10-min sieving time with sieve agitators and no dispersion agent, 2) 10-min sieving time with sieve agitators and dispersion agent, 3) 15-min sieving time with no sieve agitators or dispersion agent, 4) 15-min sieving time with sieve agitators and no dispersion agent, and 5) 15-min sieving time with sieve agitators and dispersion agent conducted in 3 grain types (ground corn, sorghum, and wheat) with 4 replicates per treatment. The analytical method that resulted in the lowest dgw and greatest Sgw was considered desirable because it was presumably representative of increased movement of particles to their appropriate sieve. Analytical method affected dgw and Sgw (P 0.05), but wheat ground using the same mill parameters was 120 to 104 m larger (P = 0.05) than corn and sorghum, respectively. Both sieve agitators and dispersion agent should be included when conducting particle size analysis. The results indicate that 10 and 15 min of sieving time produced similar results

Lois et règlements sur la faune sauvage à Madagascar : Progrès accomplis et besoins du futur

In many countries wildlife species are threatened by hunting for meat or collection for the pet trade. Wildlife laws which control where these activities can occur, limit the timing of exploitation, or provide strict protection for some species are therefore an important component of the conservation strategy. However it is important that these wildlife laws reflect the ecology and threat status of the species concerned, and that they are aligned with any relevant international conventions. In this article we discuss the legal framework for exploiting and protecting tetrapod species in Madagascar. We review the 2006 update to wildlife legislation with respect to international treaties, other national legislation and the IUCN Red List of Threatened Species. We also present a summary of the different categories of hunting (sport, commercial, scientific, and subsistence) and the control of hunting in protected areas. Madagascar has a sound legal framework for the use and protection of wildlife and the classification of species into protected, pest and legally hunted is clear and mostly fits well with the species’ classification according to the IUCN Red List and CITES. A revision of the protected species list managed is needed however to (i) include marine mammals that are protected by fisheries law and the Convention on Migratory Species and to (ii) better reflect the rights of people whose livelihoods rely heavily on the income or protein derived from hunting animals. Renewed effort to communicate and enforce wildlife legislation is needed, especially regarding the illegal hunting and export of protected species. This would also support the ongoing initiative to expand the protected area system and could be integrated into a revised National Biodiversity Strategy and Action Plan that Madagascar should produce for 2011-2020 as part of its commitment to implementing the Convention on Biological Diversity. RÉSUMÉLes lois et règlements déterminant les niveaux de protection des espèces de la faune et de la flore sauvages sont des indicateurs importants de l’importance qu’accorde un pays à la conservation de sa biodiversité. Dans cette revue, nous évaluons la cohérence entre les lois et règlements portant sur la gestion de la faune sauvage à Madagascar, en considérant la législation nationale, les conventions internationales ratifiées et la Liste Rouge de l’UICN pour les confronter aux réalités locales. Suite à nos analyses, nous pouvons conclure que Madagascar dispose d’un cadre juridique adéquat pour réglementer la protection et l’exploitation des animaux sauvages. Cependant, des révisions et mises à jour sont nécessaires, particulièrement en ce qui concerne la liste des espèces dans les différentes catégories et la facilitation de la mise en application de la loi

Efficacy of salmeterol/fluticasone propionate by GOLD stage of chronic obstructive pulmonary disease: analysis from the randomised, placebo-controlled TORCH study

Background: The efficacy of inhaled salmeterol plus fluticasone propionate (SFC) in patients with severe or very severe COPD is well documented. However, there are only limited data about the influence of GOLD severity staging on the effectiveness of SFC, particularly in patients with milder disease. Methods: TORCH was a 3-year, double-blind, placebo-controlled trial of 6112 patients with moderate/severe COPD with pre-bronchodilator FEV1 < 60% predicted (mean age 65 years, 76% male, mean 44% predicted FEV1, 43% current smokers). To understand the relative efficacy of SFC and its components by GOLD stages, we conducted a post-hoc analysis of the TORCH dataset using baseline post-bronchodilator FEV1 to segment patients into three groups: moderate COPD (GOLD stage II and above: ³ 50%; n = 2156), severe COPD (GOLD stage III: 30% to < 50%; n = 3019) and very severe COPD (GOLD stage IV: < 30%; n = 937). Results: Compared with placebo, SFC improved post-bronchodilator FEV1: 101 ml (95% confidence interval [CI]: 71, 132) in GOLD stage II, 82 ml (95% CI: 60, 104) in GOLD stage III and 96 ml (95% CI: 54, 138) in GOLD stage IV patients, and reduced the rate of exacerbations: 31% (95% CI: 19, 40) in GOLD stage II, 26% (95% CI: 17, 34) in GOLD stage III and 14% (95% CI: -4,29) in GOLD stage IV. SFC improved health status to a greater extent than other treatments regardless of baseline GOLD stage. Similarly, SFC reduced the risk of death by 33% (hazard ratio [HR] 0.67; 95% CI: 0.45, 0.98) for GOLD stage II, 5% (HR 0.95; 95% CI: 0.73, 1.24) for GOLD stage III, and 30% (HR 0.70; 95% CI: 0.47, 1.05) for GOLD stage IV. The rates of adverse events were similar across treatment arms and increased with disease severity. Overall, there was a higher incidence of pneumonia in the fluticasone propionate and SFC arms, compared with other treatments in all GOLD stages. Conclusion: In the TORCH study, SFC reduced moderate-to-severe exacerbations and improved health status and FEV1 across GOLD stages. Treatment with SFC may be associated with reduced mortality compared with placebo in patients with GOLD stage II disease. The effects were similar to those reported for the study as a whole. Thus, SFC is an effective treatment option for patients with GOLD stage II COPD

Tissue memory CD4+ T cells expressing IL-7 receptor-alpha (CD127) preferentially support latent HIV-1 infection.

The primary reservoir for HIV is within memory CD4+ T cells residing within tissues, yet the features that make some of these cells more susceptible than others to infection by HIV is not well understood. Recent studies demonstrated that CCR5-tropic HIV-1 efficiently enters tissue-derived memory CD4+ T cells expressing CD127, the alpha chain of the IL7 receptor, but rarely completes the replication cycle. We now demonstrate that the inability of HIV to replicate in these CD127-expressing cells is not due to post-entry restriction by SAMHD1. Rather, relative to other memory T cell subsets, these cells are highly prone to undergoing latent infection with HIV, as revealed by the high levels of integrated HIV DNA in these cells. Host gene expression profiling revealed that CD127-expressing memory CD4+ T cells are phenotypically distinct from other tissue memory CD4+ T cells, and are defined by a quiescent state with diminished NFκB, NFAT, and Ox40 signaling. However, latently-infected CD127+ cells harbored unspliced HIV transcripts and stimulation of these cells with anti-CD3/CD28 reversed latency. These findings identify a novel subset of memory CD4+ T cells found in tissue and not in blood that are preferentially targeted for latent infection by HIV, and may serve as an important reservoir to target for HIV eradication efforts