Can the problem of hybridization in threatened species be evaluated using a fieldwork research? A case study in snapdragons

Hybridization, natural or artificial, is considered disadvantageous for species biodiversity when it threatens the population integrity of endangered species. Frequently, studies investigating whether hybridization poses a legitimate risk to rare species are based on genetic data obtained in molecular biology laboratories. In this study, we used field research to approach the problem that hybridization could cause for the viability of a population of a rare species and to be able to propose the most appropriate initial conservation strategy. Specifically, using the model genus Antirrhinum, the reproductive barriers between the rare A. pulverulentum and its common congener A. litigiosum have been analysed under the reproductive isolation index (RI). A. pulverulentum had a high value of total RI, indicating that there are barriers to gene flow from A. litigiosum towards this species, and also had a high value for the intrinsic RI, reflecting a low inherent capacity for production of hybrid plants; in addition, the possibility of successful backcrosses between this species with hybrids produced from A. litigiosum ovules were low, given the high intrinsic RI of A. litigiosum. These data indicate the current existence of strong and permanent barriers to hybridization between the two species, suggesting that hybridization does not seem to be a serious problem for the conservation of A. pulverulentum in the studied population, nor for the near future. This study shows how the RI index can provide useful information for conservation purposes and proposes different management recommendations.     

An assessment of the AFLP method for investigating population structure in the red alga Chondrus crispus Stackhouse (Gigartinales, Florideophyceae)

The appropriateness of the Amplified Fragment Length Polymorphism (AFLP) technique for investigating Chondrus crispus Stackhouse populations in the Maritime Provinces of Canada was assessed. The AFLP procedure was first subjected to reproducibility testing and three shortcomings were noted: 1) failure to reproduce band intensity between replicate runs for the same individual and primer pair; 2) failure of some bands to replicate; 3) lack of reproducibility for complete replicate runs for some individuals and primer pairs. In the last-mentioned case, the lack of reproducibility resulted in characteristic electropherograms indicative of weak reactions. These weak runs can be attributed to poor restriction digest/ligation reactions and/or substandard PCR, these failures ultimately resulting from low and inconsistent DNA quality. We recommend that reproducibility testing should be completed routinely in studies using the AFLP technique. In the current work, only fragments and individuals that gave reproducible results were used in subsequent analyses. The AFLP method resulted in highly variable markers within and between the populations of C. crispus included in this investigation, which prevented successful resolution of population structure. This situation could result from a lack of suitability for AFLP markers in population genetic studies, and/or too extensive genetic variation for C. crispus populations to be discerned by the AFLP technique. These two possible explanations are discussed. This revised version was published online in August 2006 with corrections to the Cover Date.     

Should biochemical markers of bone turnover be considered standard practice for safety pharmacology?

The success in biomedical sciences such as genomics and proteomics is not paralleled in the medical product development methods. The consequence of this is a lack of translation into improved drug safety and efficacy. Therefore the US Food and Drug Administration (FDA) introduced the Critical Path Initiative in 2004 to modernize drug development and safety pharmacology. Bone is that largest tissue by weight, and is continuously remodelled. Changes in bone turnover lead to complications such as osteoporosis and fracture, that is associated with an increased mortality. Recent findings have identified bone as a possible endocrine organ and the availability of valid biochemical bone markers suggests that assessing bone turnover should also play an important role in general safety pharmacology.     

A rare hybridization event in two common Caribbean wrasses (genus <Emphasis Type="Italic">Halichoeres</Emphasis>; family Labridae)

Molecular tools were used to evaluate the hybrid status of a specimen with intermediate colour pattern between Halichoeres bivittatus and Halichoeres garnoti from Belize. Phylogenetic analyses of the two species, eight Halichoeres species from new and old world lineages and two outgroups showed that the study species are closely related and that H. garnoti is the maternal contributor to the putative hybrid specimen, based on partial mitochondrial COI data. Direct sequencing of Intron 1 of the nuclear ribosomal protein S7 identified H. bivittatus as sister to H. garnoti with the putative hybrid specimen in an intermediate position, due to heterozygosity at nucleotides alternatively fixed in the two putative parent species. This is consistent with the hybrid status of the specimen, with parental contributions from both H. garnoti and H. bivittatus. These results, combined with no evidence of introgression between the two parent species (based on the mtDNA and the single investigated nuclear marker) and the biogeography and ecology of these species suggests that this is a rare event with minimal evolutionary implications.     

中老年健康体检人群多项肿瘤标志物筛查结果及与年龄和性别的关系分析

摘要 目的：研究中老年健康体检人群多项肿瘤标志物（TM）筛查结果,分析其与年龄和性别的关系。方法：将自2017年6月至2019年12月于我院接受健康体检的600例中老年体检者纳入研究,分别以流式荧光发光法检测甲胎蛋白（AFP）、癌胚抗原（CEA）、糖类抗原-153（CA-153）、糖类抗原-125（CA-125）水平,并分析不同性别、年龄的体检者上述TM阳性率情况,分析各项TM阳性率检出疾病情况。结果：600例体检者AFP、CEA、CA-153、CA-125水平分别为（17.48±3.84）ng/mL、（4.54±1.19）ng/mL、（29.23±7.10）U/mL、（30.65±6.39）U/mL,阳性率分别为1.67%、4.00%、3.33%、3.83%。男性体检者CEA及CA-153阳性率均高于女性,而CA-125阳性率低于女性（P＜0.05）;男性与女性AFP阳性率对比不明显（P＞0.05）。81～89岁体检者的AFP、CEA、CA-153、CA-125阳性率均高于其他年龄段体检者（P＜0.05）。AFP、CEA、CA-153、CA-125阳性体检者恶性肿瘤检出率较低。结论：AFP、CEA、CA-153、CA-125表达升高,可能是中老年人群肿瘤发生、发展的早期预警,应予以高度重视,且和年龄、性别关系密切。中老年人群定期体检能够早期发现疾病并治疗,防止疾病继续恶化,保证身心健康。     

Nimbolide prevents myocardial damage by regulating cardiac biomarkers,antioxidant level,and apoptosis signaling against doxorubicin‐induced cardiotoxicity in rats

The current work planned to assess the protecting properties of nimbolide against doxorubicin (DOX)‐treated myocardial damage. Myocardial damage was produced with 2.5 mg/kg of DOX given on alternative days (14 days). Thiobarbituric acid reactive substances (TBARS) levels of a lipid peroxidative marker were elevated, whereas reduced body weight, heart weight, blood pressure indices and reduced levels of antioxidants like glutathione‐S‐transferase, superoxide dismutase, catalase, glutathione peroxidase, glutathione, and glutathione reductase were observed in the heart tissue of DOX‐treated animals. DOX‐treated animals showed augmented levels of cardiac markers likes monocyte chemotactic protein‐1, interferon‐gamma, aspartate transferase, creatine kinase, lactate dehydrogenase, creatine kinase‐muscle/brain, heart‐type fatty acid‐binding protein, glycogen phosphorylase isoenzyme BB, transforming growth factor‐β, brain natriuretic peptide, myoglobin, and cTnI in serum. Histopathological assessment confirmed the DOX‐induced cardiotoxicity. Furthermore, DOX‐induced rats showed augmented inflammatory mediators (nuclear factor‐κB [NF‐kB], tumor necrosis factor‐α [TNF‐α], and interleukin‐1β [IL‐1β]) and increased PI3K/Akt signaling proteins (PI3K, p‐Bad/Bad, caspase‐3, and p‐Akt), whereas decreased oxidative markers (HO‐1 and NQO‐1) and p‐PTEN were observed. Nimbolide‐supplemented rats showed reduced activity/levels of cardiac markers and TBARS levels in serum and heart tissue. Levels of enzymatic and nonenzymatic antioxidants were augmented in the heart tissue of nimbolide‐supplemented rats. Nimbolide influence decreased apoptosis, inflammation, and enhanced antioxidant markers through the modulation of p‐Bad/Bad, caspase‐3, PI3K, p‐Akt, TNF‐α, NF‐kB, IL‐1β, HO‐1, NQO‐1, and p‐PTEN markers. The histopathological explanations were observed to be in line with biochemical analysis. Therefore, the finding of current work was that nimbolide has a defensive effect on the myocardium against DOX‐induced cardiac tissue damage.     

Mouse Sertoli cells isolation by lineage tracing and sorting

Sertoli cells play a key role in spermatogenesis by supporting the germ cells throughout differentiation. The isolation of Sertoli cells is essential to study their functions. However, the close contact of Sertoli cells with other testicular cell types and the high proliferation of contaminating cells are obstacles to obtain pure primary cultures. Current rodent Sertoli cell isolation protocols result in enriched, rather than pure Sertoli cells. Therefore, novel approaches are necessary to improve the purity of Sertoli cell primary cultures. The goal of this study is to obtain pure mouse Sertoli cells using lineage tracing and fluorescence‐activated cell sorting (FACS). We bred the Amh‐Cre mouse line with tdTomato line to generate mice constitutively expressing red fluorescence specifically in Sertoli cells. Primary cultures of Sertoli cells isolated from prepubertal mice showed that 79% of cells expressed tdTomato, as evaluated by fluorescence microscopy and flow cytometry; however, nearly all adherent cells were positive for vimentin. Most of the tomato‐negative cells expressed α‐smooth muscle actin (α‐SMA), a peritubular myoid cell marker, but double‐negative populations were also present. These findings suggest that vimentin lacks Sertoli cell‐specificity and that α‐SMA is not adequate to identify all of the contaminating cells. Upon FACS sorting; however, virtually 100% of the cells were tdTomato positive, expressed vimentin, but not α‐SMA. Prepubertal mice yielded a higher number of Sertoli cells compared to adults, but both could be adequately sorted. In conclusion, our study shows that lineage tracing and sorting is an efficient strategy for acquiring pure populations of murine Sertoli cells.     

分子标记在濒危物种保护中的应用

分子标记可揭示种群遗传和进化信息, 为制定濒危物种保护措施、指导恢复实践提供重要依据。本文主要介绍了分子标记在濒危物种保护过程不同环节中的应用, 包括: (1)正确识别保护单元, 如排除隐存种和杂交种的影响; (2)确定优先保护单元, 包括优先保护区域、优先保护物种、优先保护种群等; (3)指导迁地保护; (4)对保护工作的动态监测和评估。文章最后探讨了分子标记应用于保护的发展方向, 如开展长期的种群遗传组成监测、切实应用于保护管理实践、将基因组学等遗传信息用于全球变化背景下保护策略的制定等, 期望为分子标记技术在生物多样性保护的研究和实践中提供参考。