Effect of semen preparation technique and its incubation on sperm quality in the Moroccan population

In in vitro fertilisation (IVF), sperm preparation as critical part and influencing the sperm quality is especially dependent on the chosen technique itself and incubation parameters including temperature and CO2. In this study, we compared firstly density‐gradient centrifugation technique (DGC) to the adapted DGC using the sperm pellet of 80% fraction (DGC/80P) in order to improve the sperm yield. Secondly, this study led to evaluate different sperm incubation conditions based on temperature effect (room temperature (RT = 23°C) versus 35°C) and in the other hand, with or without 5% CO2 during 24 hrs. Based on evaluating sperm conventional parameters and the DNA damage using TUNEL assay, our result showed that DGC/80P increased sperm quality compared to DGC with 25% of improvement. For temperature incubation effect after 24 hrs, 35°C increased the DNA damage and decreased the sperm quality while RT could improve sperm motility by 38%. Moreover, the sperm incubation with 5% CO2 after 24 hrs realised a negative impact on sperm parameters and its DNA damage. Indeed, for current IVF practice, a good sperm quality can be maintained for several hours at room temperature, while the sperm preparation is processed using the DGC/80P without CO2.     

Nuclear DNA replication and repair in parasites of the genus Leishmania: Exploiting differences to develop innovative therapeutic approaches

Leishmaniasis is a common tropical disease that affects mainly poor people in underdeveloped and developing countries. This largely neglected infection is caused by Leishmania spp, a parasite from the Trypanosomatidae family. This parasitic disease has different clinical manifestations, ranging from localized cutaneous to more harmful visceral forms. The main limitations of the current treatments are their high cost, toxicity, lack of specificity, and long duration. Efforts to improve treatments are necessary to deal with this infectious disease. Many approved drugs to combat diseases as diverse as cancer, bacterial, or viral infections take advantage of specific features of the causing agent or of the disease. Recent evidence indicates that the specific characteristics of the Trypanosomatidae replication and repair machineries could be used as possible targets for the development of new treatments. Here, we review in detail the molecular mechanisms of DNA replication and repair regulation in trypanosomatids of the genus Leishmania and the drugs that could be useful against this disease.     

Genetic classification of Echinococcus granulosus cysts from humans, cattle and camels in Libya using mutation scanning-based analysis of mitochondrial loci

We genetically classified Echinococcus granulosus from humans, cattle and camels in Libya utilizing DNA regions (designated pcox1 and pnad1) within the cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase 1 (nad1) mitochondrial genes, respectively. Polymerase chain reaction (PCR)-based single-strand conformation polymorphism (SSCP) analysis of pcox1 and pnad1 amplicons derived from genomic DNA samples from individual cysts (n = 176) revealed four distinct electrophoretic profiles for each locus. Direct sequencing of selected amplicons representing each of these profiles defined four different sequence types for each locus, which were present in five different combinations (designated haplotypes A–E) amongst all 176 isolates. Phylogenetic analysis of concatenated sequence data for these five haplotypes, together with a range of well-defined reference sequences, inferred that all cyst isolates from humans (n = 55) and a small number from cattle (13% of 38) belonged to the G1–G3 complex of E. granulosus (or E. granulosus sensu stricto), whereas most (87%) cysts from cattle and all 83 of them from camels were linked to the G6–G10 complex (or Echinococcus canadensis). The present study provides a foundation for future large-scale studies of the epidemiology and ecology of E. granulosus in Libya and other African countries.     

Case of localized scleroderma associated with osteomyelitis

We report a 4-year-old girl presenting with progressive linear scleroderma affecting the right leg. Biopsy specimen disclosed typical histopathological findings of localized scleroderma. Right leg magnetic resonance imaging (MRI) showed high signal areas on T₂-weighted images on the subcutaneous fatty tissue, muscles and bone marrow, suggesting that skin inflammation extended to the bone marrow. Oral corticosteroid therapy was instituted with improvement of both skin sclerosis and MRI findings. Our observations suggest that MRI examination should be considered in patients with localized scleroderma to evaluate the extension of the inflammation.     

Insight into the mechanisms underlying tumor response to boron neutron capture therapy in the hamster cheek pouch oral cancer model

Objective: The therapeutic success of different boron neutron capture therapy (BNCT) protocols employing the hamster cheek pouch oral cancer model has been previously reported by our laboratory. The aim of this study was to explore potential mechanisms of BNCT‐induced damage to tumor in terms of potential inhibition in DNA synthesis and induction of apoptosis in the tumors that underwent partial remission following application of the different BNCT protocols in this model. Materials and methods: We evaluated DNA synthesis employing incorporation of 5‐bromo‐2′‐deoxyuridine as an end‐point. Apoptosis was evaluated by immunohistochemistry employing the deoxynucleotidyl transferase‐mediated deoxyuridine triphosphate‐biotin nick end‐labeling technique and Bax and Bcl‐2 labeling. These studies were performed in tumors that underwent partial remission 1–30 days post‐BNCT mediated by boronophenylalanine (BPA), GB‐10 (Na₂¹⁰B₁₀H₁₀) or (BPA + GB‐10). Results: BNCT exerted a marked inhibitory effect on DNA synthesis in tumors for all the protocols under study. The inhibitory effect of BPA‐BNCT occurred as soon as 1 day post‐treatment (P < 0.001). Conversely, the effect of GB‐10‐BNCT became apparent 7–14 days after therapy (P < 0.001) and was sustained until killed at 30 days post‐treatment (P < 0.001). (GB‐10 + BPA)‐BNCT exerted a rapid and persistent effect, conceivably because of the combined effect of BNCT mediated by both boron compounds. The apoptosis studies did not show differences between the pre‐treatment group and any of the BNCT groups. Conclusions: One of the mechanisms involved in BNCT‐induced tumor control in our model would be an inhibitory effect on DNA synthesis. Apoptosis does not seem to have a significant role in BNCT‐induced tumor control in our model.     

乙型肝炎病毒DNA与标本物含量及肝功能生化指标间关系探讨

目的探讨血清中乙型肝炎病毒(HBV)DNA含量与HBV标志物[乙型肝炎表面抗原(HBsAg)、乙型肝炎e抗原(HBeAg)]及肝功能指标间的相关性。方法抽取南平市人民医院肝内科住院的200例HBV携带患者血清样本。采用荧光定量聚合酶链反应(PCR)检测血清样本中HBV DNA含量,采用酶联免疫吸附试验(ELISA)测定血清样本中HBV标志物含量,同时检测肝功能。3项检测指标使用同一患者标本,分析3项指标之间的相关性。结果经相关性分析,HBV DNA与肝功能指标的r在±0.2以内,差异无统计学意义(P>0.05);HBsAg、HBeAg含量与肝功能指标的r在±0.2以内,差异无统计学意义(P>0.05);HBsAg、HBeAg含量与HBVDNA含量的r分别为0.457、0.568,差异有统计学意义(P<0.01)。结论 HBsAg、HBeAg含量只反映HBV在血清中的复制情况,不能以HBsAg、HBeAg含量及病毒载量的大小作为判断肝功能受损程度的指标;要综合分析HBV DNA含量与HBV标志物定量检测结果及肝功能,这样才能更有效地对疾病进行评估。     

Characterization and distribution of retrotransposons and simple sequence repeats in the bovine genome

Interspersed repeat composition and distribution in mammals have been best characterized in the human and mouse genomes. The bovine genome contains typical eutherian mammal repeats, but also has a significant number of long interspersed nuclear element RTE (BovB) elements proposed to have been horizontally transferred from squamata. Our analysis of the BovB repeats has indicated that only a few of them are currently likely to retrotranspose in cattle. However, bovine L1 repeats (L1 BT) have many likely active copies. Comparison of substitution rates for BovB and L1 BT indicates that L1 BT is a younger repeat family than BovB. In contrast to mouse and human, L1 occurrence is not negatively correlated with G+C content. However, BovB, Bov A2, ART2A, and Bov-tA are negatively correlated with G+C, although Bov-tAs correlation is weaker. Also, by performing genome wide correlation analysis of interspersed and simple sequence repeats, we have identified genome territories by repeat content that appear to define ancestral vs. ruminant-specific genomic regions. These ancestral regions, enriched with L2 and MIR repeats, are largely conserved between bovine and human.     

线粒体DNAl6189T→C变异与2型糖尿病发病机制的关系

目的　探讨线粒体DNA16189T→C变异与 2型糖尿病 (T2DM )的关系。　方法　运用PCR -RFLP技术在 3 0 8例无血缘关系的上海地区汉族人 (糖耐量正常 158名 ,T2DM患者 150例 )中检测线粒体DNA16189T→C变异。　结果　上海地区汉族人线粒体DNA16189T→C变异频率显著高于英国白种人 (2 9%vs 9.9% ,P =6.85× 10 - 6 )。线粒体DNA16189T→C变异阳性的糖尿病 (DM )患者胰岛素抵抗 (HOMA IR)值明显低于阴性者 (P =0 .0 2 7) ,并且 ,逐步多元回归分析显示该变异是HOMA IR的独立相关因素之一 (部分R2 =0 .0 2 8,P =0 .0 2 6)。　结论　线粒体DNA16189T→C变异与上海地区汉族人T2DM胰岛素抵抗相关