A Simple Staining Procedure for Detecting the true Acrosome Reaction in Buffalo (Bubalus bubalis) Spermatozoa

A simple dual staining procedure for detecting the true acrosome reaction in dried smears of buffalo spermatozoa is described. Trypan blue is used first to differentiate live from dead spermatozoa and the dried smears which have been prepared are stained with Giemsa for acrosome evaluation. Four categories of spermatozoa were recognized: A) live, intact acrosome (acrosome pink, postnuclear cap clear); B) dead, intact acrosome (acrosome pink, postnuclear cap blue); C) live, detached acrosome (acrosome clear, postnuclear cap clear); and D) dead, detached acrosome (acrosome clear, postnuclear cap blue). The procedure is simple, rapid and convenient for assessing true acrosome reaction in buffalo spermatozoa. Simultaneous assessment of sperm viability and its acrosomal status in dried smears makes this procedure attractive because the true acrosome reaction can be studied thoroughly at a later state after the incubation period.     

Acrosome reaction in sperm of the toad,bufo bufo japonicus

The acrosome in the sperm of the toad, Bufo bufo japonicus, consists of a membrane-limited acrosomal cap and a fibrous perforatorium. When sperm are incubated with the oviducal pars recta extract (PRE) for 30–60 min, the outer acrosomal membrane fuses with the overlying plasma membrane at several points with concomitant loss of the contents of the acrosomal cap. The inner acrosomal membrane thus exposed fuses with the plasma membrane at the caudal end of the acrosomal region. This PRE-induced acrosome reaction is completely inhibited by soybean trypsin inhibitor. Sperm found in the innermost jelly layer of inseminated eggs possess an intact acrosome, but those either passing through the vitelline coat or localizing in the perivitelline space are acrosome-reacted in the same manner as when treated with PRE. These observations, combined with recent evidence showing involvement of the pars recta substance in fertilization, indicate that the acrosome reaction occurring in a fertilizing sperm at or near the surface of the vitelline coat is a response to a substance that is derived from the pars recta and deposited in the vitelline coat.     

Neural Correlates of Body Integrity Dysphoria

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Base excision repair but not DNA double‐strand break repair is impaired in aged human adipose‐derived stem cells

The decline in DNA repair capacity contributes to the age‐associated decrease in genome integrity in somatic cells of different species. However, due to the lack of clinical samples and appropriate tools for studying DNA repair, whether and how age‐associated changes in DNA repair result in a loss of genome integrity of human adult stem cells remains incompletely characterized. Here, we isolated 20 eyelid adipose‐derived stem cell (ADSC) lines from healthy individuals (young: 10 donors with ages ranging 17–25 years; old: 10 donors with ages ranging 50–59 years). Using these cell lines, we systematically compared the efficiency of base excision repair (BER) and two DNA double‐strand break (DSB) repair pathways—nonhomologous end joining (NHEJ) and homologous recombination (HR)—between the young and old groups. Surprisingly, we found that the efficiency of BER but not NHEJ or HR is impaired in aged human ADSCs, which is in contrast to previous findings that DSB repair declines with age in human fibroblasts. We also demonstrated that BER efficiency is negatively associated with tail moment, which reflects a loss of genome integrity in human ADSCs. Mechanistic studies indicated that at the protein level XRCC1, but not other BER factors, exhibited age‐associated decline. Overexpression of XRCC1 reversed the decline of BER efficiency and genome integrity, indicating that XRCC1 is a potential therapeutic target for stabilizing genomes in aged ADSCs.     

What is the value of wild animal welfare for restoration ecology?

The restoration community continues to discuss what constitutes good environmental stewardship. One area of tension is the extent to which the well‐being of wild animals should inform restoration efforts. We discuss three ways that the perspective of wild animal welfare can augment restoration ecology: strengthening people's relationship with nature, reinforcing biotic integrity, and reducing mechanistic uncertainty. The animal welfare movement elevates sentient animals as stakeholders and explores how environmental context directly impacts the well‐being of individuals. Viewing wild animals through this lens may encourage people to think and act with empathy and altruism. Second, we incorporate animal welfare into the concept of biotic integrity for ecological and ethical reasons. Restoring ecosystem processes may enhance animal welfare, and vice versa. Alternatively, there may be a trade‐off between these factors, requiring local decision‐makers to prioritize between restoring ecosystem function and promoting individuals' well‐being. We conclude by discussing how welfare can impact population recovery, thereby adding insights about mechanisms underpinning restoration objectives. Ultimately, restoration ecologists and proponents of wild animal welfare could enjoy a productive union.     

Effect of membrane integrity on survival competition of Botrytis cinerea upon QoI fungicide pyraclostrobin

Botrytis cinerea, the causal agent of the grey mould disease, developed resistance to multiple fungicides. However, the role of cell membrane in survival competition of B. cinerea upon quinone outside inhibitor (QoI) fungicide has not yet been elucidated. In this paper, the enhancement of cystamine, a transglutaminase inhibitor, on membrane integrity of B. cinerea was determined, and the effect of the enhancement on the sensitivity of B. cinerea to pyraclostrobin was investigated. The results showed that pyraclostrobin inhibited mycelial growth with EC₅₀ as 1.122 and 3.042 μg/ml at 24 and 48 hr, respectively. In the treatment of 5 and 50 μg/ml pyraclostrobin, membrane integrity of B. cinerea was broken, causing high permeability, lipid peroxidation, flocculent and malformed surface with vague septum and abundant agglomerates inside and outside the mycelia. Cystamine even at 50 and 200 μg/ml had little inhibitory effect on mycelial growth. However, in presence of 50 or 200 μg/ml cystamine, the mycelia from pyraclostrobin treatment possessed a significantly reduced leakage, lower MDA content, and a revived fibrous and transparent surface. Meanwhile, SEM images showed that membrane integrity of the mycelia was significantly improved and the agglomerates were dramatically disappeared. Synergy assays further revealed that B. cinerea regained less sensitivity to pyraclostrobin inhibition. In conclusion, membrane integrity controls mycelia sensitivity and is required for survival competition of B. cinerea upon pyraclostrobin.     

国家公园陆地自然生态系统完整性与原真性评价方法探索: 以钱江源国家公园体制试点为例

中国建立国家公园的目的是保护自然生态系统的完整性和原真性, 促进生物多样性保护。国家公园的完整性和原真性评价是国家公园的布局规划、边界范围确定以及功能区划等研究的前提条件。为了评估国家公园自然生态系统完整性和原真性状态, 本文基于陆地自然生态系统的结构和功能, 通过指标筛选、专家咨询、指标量化和建立综合评价模型, 构建了陆地自然生态系统完整性与原真性的评价指标体系及其量化评价方法。该评价方法包括5个自然生态系统完整性指标、5个自然生态系统原真性指标和2个综合评价指标。以浙江省钱江源国家公园体制试点为例, 本文初步评估了其生态系统完整性与原真性状态, 并对评价结果进行了分级。按照本研究的评价方法, 钱江源国家公园体制试点的自然生态系统完整性评价结果为52.83%, 评价等级为较差; 自然生态系统原真性评价结果为87.06%, 评价等级为好。钱江源国家公园体制试点有待关注和提升的指标有保护区域完整性指数(27.00%)和旗舰种适宜生境完整性指数(53.04%)。最后, 本文结合研究区域评价结果对生态系统完整性和原真性领域应关注的问题进行了讨论。该自然生态系统完整性和原真性评价方法可提供一种评价指标覆盖较全面、数据易获取, 且评价结果易被决策者和管理者理解的评价思路。