Analysis of sperm nuclear protein gene polymorphisms and DNA integrity in infertile men

Sperm nuclear proteins, the protamines (PRM) and transition nuclear proteins (TNP) play a crucial role in sperm nuclear condensation. The compact packaging of sperm DNA by protamines maintains sperm genome integrity, which is prerequisite for normal sperm function. However the effect of nucleotide variations in PRM and TNP genes on sperm DNA integrity and male fertility is not clear. This case-control study was planned to analyze PRM and TNP gene nucleotide variations and sperm DNA integrity in 100 oligozoospermic infertile men and 100 fertile controls. Protamine and TNP genes were amplified by polymerase chain reaction and sequenced. Flow cytometry-sperm chromatin structure assay (FC-SCSA) was applied to measure the DNA fragmentation index (DFI) in sperm. Semen analysis was performed as per WHO [1999] guidelines with slight modification. In total, 7 nucleotide variations including two novel changes, a non-synonymous mutation in the exon-2 of PRM2 gene (c.443C?>?A) and a novel insertion of T (c.396_397InsT) at the 3' UTR region of TNP1 were detected. None of the nucleotide changes were observed with increased risk frequency in the oligozoospermic infertile men compared to the controls. Though overall DFI was significantly (p??A and TNP1 c.396_397InsT had higher DFI value of 34.82% and 43.85%, respectively. In conclusion, our pilot study for the first time in the Indian population revealed two rare novel mutations in sperm nuclear protein genes that are perhaps associated with higher sperm DNA fragmentation.     

少、弱精子症患者精浆、精子和血清中锌及性激素水平的相关分析

目的:检测弱精子症和少弱精子症患者精浆、精子、血清中锌含量和血清性激素水平,分析锌含量和血清性激素水平的变化与精子密度和精子运动能力的关系。方法:随机筛选190例弱精子症患者,85例少弱精子症患者以及32例精液质量正常的已生育男性作为研究对象,利用原子吸收光谱法检测其精浆、精子和血清锌含量,放射免疫分析法检测血清生殖激素(FSH、LH、T),并进行统计学分析。结果:3组间精浆锌和血清锌含量没有显著差异(P>0.05);少弱精子症患者精子锌含量显著高于弱精子症患者和正常生育男性(P<0.01);弱精子症患者T含量显著低于正常生育组而LH含量显著高于正常生育组(P<0.05);少弱精子症患者T含量显著低于正常生育组而FSH含量显著高于正常生育组(P<0.05),并且其LH含量与正常生育组间具有统计学意义(P<0.01);弱精子症、少弱精子症患者精浆锌与血清T之间无明显的相关性(P>0.05)。结论:少弱精子症患者过高的精子锌含量可能与精子运动能力下降和精子膜功能障碍相关。但其具体机制还有待进一步研究。血清生殖激素(FSH、LH、T)的测定对于弱精子症、少弱精子症病因的诊断和治疗具有一定的临床意义。     

DNA fragmentation status in patients with necrozoospermia

The aim of this study was to determine if a relationship exists between the levels of sperm DNA fragmentation and necrospermia in infertile men. Semen samples obtained from 70 men consulting for infertility evaluation were analyzed according to World Health Organization (WHO) guidelines. Patients were subdivided into three groups according to the percentage of necrotic spermatozoa: normozoospermia (<30%; n?=?20), moderate necrozoospermia (50-80%; n?=?30), and severe necrozoospermia (>80%; n?=?20). DNA fragmentation was detected by the terminal desoxynucleotidyl transferase-mediated deoxyuridine triphosphate biotin nick-end labeling (TUNEL) assay. The sperm DNA fragmentation index (DFI) was 9.28?±?2.98% in patients with a normal level of necrotic spermatozoa, 20.25?±?3.21% in patients with moderate necrozoospermia, and 35.31?±?5.25% in patients with severe necrozoospermia. There was a statistically significant increase of DNA fragmentation in the necrozoospermic group (P?<?0.01). A strong correlation was found between the degree of necrozoospermia and sperm DNA fragmentation. We concluded that patients with necrozoospermia showed a high level of DNA fragmentation compared to normozoospermic men. Severe necrozoospermia (>80%) is a predictive factor for increased sperm DNA damage.     

Analysis of sperm nuclear protein gene polymorphisms and DNA integrity in infertile men

Sperm nuclear proteins, the protamines (PRM) and transition nuclear proteins (TNP) play a crucial role in sperm nuclear condensation. The compact packaging of sperm DNA by protamines maintains sperm genome integrity, which is prerequisite for normal sperm function. However the effect of nucleotide variations in PRM and TNP genes on sperm DNA integrity and male fertility is not clear. This case-control study was planned to analyze PRM and TNP gene nucleotide variations and sperm DNA integrity in 100 oligozoospermic infertile men and 100 fertile controls. Protamine and TNP genes were amplified by polymerase chain reaction and sequenced. Flow cytometry-sperm chromatin structure assay (FC-SCSA) was applied to measure the DNA fragmentation index (DFI) in sperm. Semen analysis was performed as per WHO [1999] guidelines with slight modification. In total, 7 nucleotide variations including two novel changes, a non-synonymous mutation in the exon-2 of PRM2 gene (c.443C?>?A) and a novel insertion of T (c.396_397InsT) at the 3′ UTR region of TNP1 were detected. None of the nucleotide changes were observed with increased risk frequency in the oligozoospermic infertile men compared to the controls. Though overall DFI was significantly (p?<?0.0001) higher in infertile men compared to controls (36.31?±?7.25 vs. 26.49?±?2.78) irrespective of nucleotide changes, no such difference was observed between 100 infertile men or pooled population of 200 with and without mutations. However it was observed that two cases with novel nucleotide changes PRM2 c.443C?>?A and TNP1 c.396_397InsT had higher DFI value of 34.82% and 43.85%, respectively. In conclusion, our pilot study for the first time in the Indian population revealed two rare novel mutations in sperm nuclear protein genes that are perhaps associated with higher sperm DNA fragmentation.     

受血及献血者HIV-1外膜蛋白ENV基因序列分析

目的　了解在广东省受血及献血者中发现的HIV 1亚型的流行规律及其与国际参考株的同源性。方法　应用套式聚合酶链反应 (PCR)对 1 4例采自于广东省HIV 1抗体阳性的受血者或献血者淋巴细胞富集液的核酸样品进行扩增 ,并使用ABI377型测序仪对扩增产物测序后 ,对其ENV基因C2 V3段的核酸序列进行比较分析。结果　 1 4份血样中 ,7份为泰国B’亚型 ,与国际参考株RL4 2的距离最近 ,基因离散率为 (6 0 82± 2 6 0 7) % ,组内离散率为 (5 96 3± 2 383) % ;4份为AE重组亚型 ,与国际参考株TH 90 CM2 4 0最近 ,基因离散率为 (8 90 0± 1 830 ) % ,组内离散率为 (1 3 81 0± 1 31 7) % ;2份为 0 7 BC重组亚型 ,与国际参考株CN 97 C5 4A最近 ,基因离散率为 (4 1 5 5± 1 2 2 3) % ,组内基因距离为 6 36 % ;1份为 0 8 BC重组亚型 ,与国际参考株 97CNGX 9F最近 ,基因距离为 0 97%。结论　本次检测的广东省受血及献血者HIV 1以泰国B’亚型为主 ,也存在主要在性途径感染人群中流行的AE重组亚型和吸毒者中流行的 0 7 BC、0 8 BC重组亚型。     

山东省HIV-1流行株多基因区亚型分析

目的 了解流行于山东省境内HIV-1毒株的亚型,尤其是流行重组型毒株的种类及变异情况,分析其来源并推测其流行趋势。方法采集93份HIV-1感染者的外周静脉防凝血,提取前病毒DNA进行体外扩增,获得包膜蛋白（env）、核心蛋白（gag）、tat区基因的核酸片段,并对各基因区核苷酸进行测定和分析。结果发现在所有目标人群中共存在B、B,、C3种亚型以及CRF07-BC、CRF08-BC、CRF01-AG和CRF01-AE4种重组毒株,其中B’亚型数量最大71株（占77．2％）,CRF01-AE9株（占9．8％）,CRF07-BC、CRFOB-BC各3株（均占3．3oA）,B、C和CRF02-AG各2株（占2．2％）。3个基因区env区变异最大,gag和tat相对保守。3种优势亚型中,A／E变异最大,B’次之,B’／C最保守。结论山东省HIV-1流行毒株亚型众多。各优势毒株发生了不同程度的变异,在山东省有流行加快的趋势,应加强防控。     

Risk of HIV-1 in rural Kenya: a comparison of circumcised and uncircumcised men

BACKGROUND: Most studies that have found an association between uncircumcised status and infection with human immunodeficiency virus type 1 (HIV-1) have compared participants from various demographic backgrounds, among which the prevalence of other risk factors might have varied. We report findings from a study conducted among men within a single ethnic community in which circumcision was dictated by the religious denomination to which the men belonged. METHODS: Of the 1217 eligible men, we included in the analysis 845 who gave blood samples for HIV-1 testing and who were confirmed as either fully circumcised (n = 398) or uncircumcised (n = 447). The seroprevalence of HIV-1 was compared between the 2 groups. RESULTS: All correlates of HIV-1 prevalence that we measured were distributed similarly between circumcised and uncircumcised men. The seroprevalence of HIV-1 was 30% among the uncircumcised men and 20% among the circumcised men. Among uncircumcised men, HIV-1 seroprevalence was similar between men from circumcising denominations (31%; n = 111) and noncircumcising denominations (30%; n = 336). The crude prevalence ratio for HIV infection associated with not being circumcised was 1.5 (95% confidence interval = 1.2-2.0); and adjustment for other measured risk factors for HIV-1 infection had little impact on this result. CONCLUSION: Our study provides evidence that circumcision is associated with a reduced risk of HIV-1 infection.     

Insulin-like growth factor-1, insulin-like growth factor binding protein-3, and cardiovascular disease risk factors in young black men and white men: the CARDIA Male Hormone Study

Cross-sectional studies have found associations between components of the insulin-like growth factor (IGF) system and hypertension, total cholesterol, and low density lipoprotein cholesterol. Using partial correlation analysis and longitudinal analysis of data collected at the year 2, year 7, and year 10 examinations, the authors assessed the associations of IGF-1 and IGF binding protein-3 (IGFBP-3) with cardiovascular disease risk factors in 544 Black and 747 White male participants in the Coronary Artery Risk Development in Young Adults (CARDIA) Male Hormone Study who were aged 20-34 years at year 2 (1987-1988). There were no consistent independent associations with blood pressure. Cross-sectionally, there were some inverse associations between IGF-1 and lipid levels in White men (strongest r = -0.095 (p = 0.02) for total cholesterol at year 7) and positive associations between IGFBP-3 and lipid levels in Black and White men (for log(triglycerides), r = 0.072-0.136). Longitudinally, a 1,000-ng/ml increase in IGFBP-3 was associated with 3.7-mg/dl and 2.6-mg/dl higher total cholesterol levels and 2.6-mg/dl and 1.7-mg/dl higher low density lipoprotein cholesterol levels in Black men and White men (p < 0.05), respectively. These findings do not support a strong link between IGF-1 and IGFBP-3 and blood pressure, but they do support the possibility of important relations between IGFBP-3 and lipid levels in young adult men.     

Antibodies Against Laminin-1 and Sperm,Intraacrosomal Proteins in Semen from Infertile Couples

Seminal sperm-agglutinating antibodies along with IgG antibodies against laminin-1 and intraacrosomal sperm proteins were examined in seventy-one men from infertile couples. The direct mixed antiimmunoglobulin reaction test for IgG, IgA, and the commercial ELISA method for detecting IgG antibodies against laminin-1 in seminal plasma were used. Intraacrosomal proteins in the sperm heads were detected by immunofluorescence using monoclonal antibodies. Cellular elements other than spermatozoa, collectively referred to as “round cells” were also examined. In association with a group of oligoasthenospermatic men, positive levels (44%) of antibodies against laminin-1 of the IgG isotype in seminal plasma were found in conjunction with increased cellularity in semen. Interestingly, the elevated levels of anti-laminin-l IgG and sperm agglutinating positivity were not correlated.

The use of antibodies against sperm antigens targeted to adhesive molecules such as laminin-1 contributes to diagnosing reproductive failure. Detection of intraacrosomal proteins very often correlates to the state of semen pathology and inflammation.     

Changes over time in sexual and relationship functioning of people with multiple sclerosis

Relationship and sexual satisfaction among people with multiple sclerosis (MS) has received little research attention. This article reports on a longitudinal investigation of the impact of coping style and illness-related variables among people with MS and the general population on sexual and relationship functioning. Both men (n = 120) and women (n= 201) with MS and men (n = 79) and women (n = 160) from the general population participated in the study at two points in time, 6 months apart. People with MS experienced lower levels of sexual activity, sexual satisfaction, and relationship satisfaction and higher levels of sexual dysfunction. Generally speaking, the time 1 levels of the health and coping variables explained little of the variance in the time 2 sexual and relationship variables. However, sexual activity at time 1 contributed significant unique variance to relationship satisfaction at time 2 for MS men, and coping strategies at time I contributed to relationship satisfaction among general population women at time 2. Furthermore, for those respondents who had been diagnosed with MS for less than 7 years, levels of sexual activity at time 2 were predicted by levels of sexual and relationship satisfaction, as well as levels of sexual activity at time 1. These results suggest that strategies used to cope with illness may not play a major role in sexual and relationship satisfaction. However, an examination of these strategies over a longer time frame is needed.     

Airway inflammation in paper mill workers

Paper mill workers are exposed to culturable microorganisms (MOs). We hypothesized that inflammatory airway response could be detected in sputum of nonsymptomatic workers. From four paper mills, we included 29 healthy nonsmoking men. Workers exposed to high levels of MOs (HMOE, n = 17) were compared with workers exposed to low levels of MO (LMOE, n = 12). A reference group of 22 healthy, nonsmoking, nonexposed (NE) men were also included. We performed differential cell counts of induced sputum, studied gene expressions of isolated sputum macrophages and analyzed inflammatory parameters, including matrix metalloproteinases. Sputum from HMOE workers had a significantly higher percentage of neutrophils than that from LMOE workers (P < 0.05) and NE controls (P < 0.001). There was also an increased gene expression of different pro-inflammatory cytokines, interleukin-6, tumor necrosis factor-alpha, and macrophage inflammatory protein-1beta in isolated airway macrophages and increased levels of total matrix metalloprotease-9 activity in induced sputum from the HMOE group. Our findings indicate that paper industry workers exposed to MOs develop subclinical airway inflammation.     

Frequencies of glyoxalase I phenotypes as biological markers in chronic alcoholism

Biological markers for alcoholism would be a valuable tool for early diagnosis. We have studied the phenotype frequencies of genetically determined erythrocyte enzymes in 397 alcoholics, including two populations with liver disease: steatosis (n = 86) and cirrhosis (n = 128) and a population of alcoholics without apparent liver disease (n = 183) compared to a well selected control population (n = 177). Only for Glyoxalase I (GLO) phenotypes (1,2 and 2-1) were significant differences found between the male controls and the male alcoholics. In the total male alcoholic population the frequency of phenotype 1 was significantly increased (23.2% vs. 11%, p less than 0.02), and the frequency of phenotype 2 was significantly decreased (32.3% vs. 46.3 p less than 0.02) compared to the male control population. For normal women the frequency of phenotype 1 and 2 was significantly different from normal men. (1: +177% p less than 0.001, 2: -45% p less than 0.01), but no significant differences were found between alcoholic and normal women. Our results suggest that in male subjects Glyoxalase I phenotype 1 may provide a marker for predisposition to alcoholism.     

Impact of PCB and p,p'-DDE contaminants on human sperm Y:X chromosome ratio: studies in three European populations and the Inuit population in Greenland

OBJECTIVE: Recent studies indicate that persistent organohalogen pollutants (POPs) may contribute to sex ratio changes in offspring of exposed populations. Our aim in the present study was to investigate whether exposure to 2,2 ,4,4 ,5,5 -hexachlorobiphenyl (PCB-153) and dichlorodiphenyldichloroethene (p,p -DDE) affects sperm Y:X chromosome distribution. SUBJECTS AND METHODS: We obtained semen and blood for analysis of PCB-153 and p,p -DDE levels from 547 men from Sweden, Greenland, Poland (Warsaw), and Ukraine (Kharkiv), with regionally different levels of POP exposure. The proportion of Y- and X-chromosome-bearing sperm in the semen samples was determined by two-color fluorescence in situ hybridization analysis. RESULTS: Swedish and Greenlandic men had on average significantly higher proportions of Y sperm (in both cohorts, 51.2%) and correspondingly higher lipid-adjusted concentrations of PCB-153 (260 ng/g and 350 ng/g, respectively) compared with men from Warsaw (50.3% and 22 ng/g) and Kharkiv (50.7% and 54 ng/g). In the Swedish cohort, log-transformed PCB-153 and log-transformed p,p -DDE variables were significantly positively associated with Y-chromosome fractions (p-values 0.04 and <0.001, respectively). On the contrary, in the Polish cohort PCB-153 correlated negatively with the proportion of Y-bearing fraction of spermatozoa (p=0.008). CONCLUSIONS: The present study indicates that POP exposure might be involved in changing the proportion of ejaculated Y-bearing spermatozoa in human populations. Intercountry differences, with different exposure situations and doses, may contribute to varying Y:X chromosome ratios.