The epigenetic alterations of human sperm cells caused by heroin use disorder.

The molecular mechanisms of drug use on sexual health are largely unknown. We investigated, the relationship between heroin use disorder and epigenetic factors influencing histone acetylation in sperm cells. The volunteers included twenty-four 20- to 50-year-old men with a normal spermogram who did not consume any drugs and twenty-four age- to BMI-matched men who consume only the drug heroin for more than last four months. HDAC1 and HDAC11 mRNA expression levels in spermatozoa and miR-34c-5p and miR-125b-5p expression levels in seminal plasma were measured. The heroin-user group showed significantly increased white blood cell counts and decreased sperm motility and survival rates (8.61 ± 1.73, 21.50 ± 3.11, 69.90 ± 4.69 respectively) as compared to the control group (1.49 ± 0.32, 38.82 ± 3.05, 87.50 ± 0.99 respectively) (p ≤ .001). An increase in DNA fragmentation index (DFI) (heroin-user group: 41.93 ± 6.59% and control group: 10.14 ± 1.43%, p = .003), a change in frequency of HDAC1 (heroin-user group: 1.69 ± 0.55 and control group: 0.45 ± 0.14, p = .045) and HDAC11 (heroin-user group: 0.29 ± 0.13 and control group: 2.36 ± 0.76, p = .019) in spermatozoa and a significant decrease in seminal miR-125b-5p abundance (heroin-user group: 0.37 ± 0.11 and control group: 1.59 ± 0.47, p = .028) were reported in heroin consumers. Heroin use can lead to male infertility by causing leukocytospermia, asthenozoospermia, DFI elevation in sperm cells and alterations in seminal RNA profile.

Underlying pathway of factors leading to mental health in Iranian young adolescents: A structural equation modeling.

BACKGROUND: The present study aimed to provide a field-tested model of constituting factors affecting mental health in young Iranian adolescents. MATERIALS AND METHODS: In this cross-sectional study, a conceptual model was proposed based on an extensive literature review. A total of 254 young adolescents aged 11-14 years were recruited from north, south, east, and west regions of Tehran megacity by a random cluster sampling procedure, of whom 244 adolescents participated. The adolescents and their mothers altogether completed eight questionnaires pertaining to the proposed conceptual model: (1) Strengths and Difficulties Questionnaire (SDQ), (2) Childhood Experience of Care and Abuse Questionnaire, (3) Child Exposure to Domestic Violence Scale, (4) Drug Abuse Screening Test-10, (5) Baumrind Parenting Style Questionnaire, (6) Conflict Behavior Questionnaire, (7) General Health Questionnaire-28, and (8) Garmaroodi Socioeconomic Status Questionnaire. The statistical analysis was performed using structural equation modeling. RESULTS: This study demonstrated that parent mental health (b = -0.111), experience of father's care (b = -1.112), conflict with mother (b = 0.309), conflict with father (b = 0.135), and exposure to domestic violence (b = 0.217), as well as age (b = 0.93) and gender (b = 0.139), had direct effect on adolescent mental health (all P < 0.05). Further, the results showed that exposure to domestic violence and conflict with mother had the greatest direct impact on adolescent mental health among all other family-related factors, followed by conflict with father and parent mental health. Conflict with mother and conflict with father also affected adolescent mental health indirectly through experience of domestic violence and had a mediating effect for the influence of several other factors on adolescent mental health, thus playing an important role in the pathway leading to young adolescent mental health status in the Iranian population. CONCLUSION: Overall, the final model proved to be fit and the factors constituting the final model were able to predict 88% of the variations in the mental health of Iranian adolescents. This model can guide clinical psychologists, psychiatrists, and other mental health workers in a more realistic and effective prevention or treatment planning for their young clients. Moreover, it may help in arriving at a comprehensive preventive policymaking for mental health policymakers.

Correlation Between Protamine-2 and miRNA-122 in Sperm from Heroin-addicted Men: A Case-Control Study.

PURPOSE: Recreational use of illicit drugs is one of the main factors affecting male fertility. However, the mechanisms of heroin smoke-associated damage to mature spermatozoa are still completely unknown. The aim of this study was to concomitantly examine the levels of protamine-2 gene and protein concentrations, the amount of miRNA-122 in seminal plasma and semen analysis findings in heroin-addicted men. MATERIALS AND METHODS: In a case control study, twenty-four fertile men that lacked any recreational drug abuse were considered as the healthy group, and 24 addicted men who used only heroin for at least four months were selected as the addicted group. Semen samples were gathered by masturbation after 2 - 5 days of sexual abstinence. Following the preparation of a semen analysis by computer-assisted sperm analysis according to WHO (2010), the level of protamine-2 gene expression in sperm and miRNA-122 in seminal plasma was measured using real-time sqPCR. Also, protamine-2 protein concentrations were quantified by nuclear protein extraction, SDS-Page and western blotting. RESULTS: Among the studied variables, body mass index (27.75±0.88 vs. 22.30±0.36, p=0.001), seminal pH (7.79±0.06 vs. 7.58±0.06, p=0.003), white blood cell count in semen (1.69±0.41 vs. 8.61±1.73, p=0.001), motility (65.51±2.57 vs. 41.96±3.58, p=0.001) and survival rate (87.41±1.00 vs. 71.50±4.59, p=0.002) of sperm cells was significantly different between the healthy and addicted groups. In addition, the levels of protamine-2 gene and protein expression in the addicted group (0.05±0.02 and 0.10±0.02, respectively) were significantly lower than the healthy group (3.59±0.94 and 0.27±0.06, respectively) (p=0.002 and p=0.017, respectively). Seminal miRNA-122 levels in addicted men (3.51±0.73) were statistically higher than in healthy men (1.52±0.54) (p=0.034). However, there were some significant relationship between the studied parameters and addiction (p<0.05). CONCLUSION: This is one study on human infertility that evaluates the effects of heroin on protamine deficiency and seminal small RNAs expression levels. Heroin abuse may lead to male infertility by causing leukocytospermia, asthenozoospermia, protamine deficiency, and seminal plasma miRNA profile alteration.

The Psychometric Properties of the Drug Abuse Screening Test.

BACKGROUND: Substance abuse is a critical problem in most countries, especially in developing ones. Early detection is the pre-requisite of early control, for which reliable and valid tools are required. In the present study, we aimed at measuring the psychometric properties of the 10-item Drug Abuse Screening Test (DAST-10) in Iranian individuals. METHODS: After translation and back-translation of the questionnaire, 244 adults were recruited from Tehran Megacity, Iran, and completed the questionnaires. Participants were recruited by a multistage randomized cluster sampling method. Reliability was determined by Cronbach's alpha. Also, construct validity was evaluated using confirmatory factor analysis (CFA) and exploratory factor analysis (EFA). FINDINGS: The internal consistency using Cronbach's alpha coefficients for the total score of the Persian version of DAST-10 was 0.93. EFA evoked only one factor for DAST-10. The CFA for 1-factor models for DAST-10 indicated an acceptable fit for the proposed models. CONCLUSION: The results prove desirable reliability and validity of the Persian version of the DAST which can be utilized as a screening instrument for drug abuse among Iranian adults.

The Psychometric Properties of the Persian Version of Childhood Experience of Care and Abuse Questionnaire (CECA).

BACKGROUND: The present study aimed to determine the psychometric properties of the Persian version of the Childhood Experience of Care and Abuse Questionnaire (CECA.Q), a tool based on a retrospective interview with the child. MATERIALS AND METHODS: To this aim, 251 adolescents from four regions of Tehran megacity completed the questionnaire. The reliability of the questionnaire was examined, along with the face and content validity. In addition, the construct validity was evaluated by exploratory factor analysis (EFA) and confirmatory factor analysis (CFA). RESULTS: EFA and CFA supported a 4-factor solution including mother's role scale items, father's role scale items, maternal behavior scale items, and paternal behavior scale items. The total variance extracted in EFA ranged from 33.9 to 60.7. The internal consistency for mother's role, father's role, maternal behavior, and paternal behavior was 0.61, 0.65, 0.86, and 0.9 respectively. Thus, the questionnaire had a suitable fit, as well as reasonable reliability and validity. CONCLUSION: The Persian version of the CECA.Q had adequate reliability and validity as a self-report measurement for childhood experience of care and abuse.

Evaluation of Enkephalin-Degrading Enzymes in Sperm from Heroin-Addicted Men.

BACKGROUND: The aim of this study was to investigate two enkephalin-degrading enzymes, aminopeptidase N (APN/ CD13) and endopeptidase (NEP/CD10), gene and protein expression levels in sperm samples of fertile and heroinaddicted men, and the correlation between their expressions and semen quality. MATERIALS AND METHODS: In this case-controlled study, semen was collected from 24 normozoospermic healthy (as a control group) and 24 heroin-addicted men donors (as case or addiction group). Sperm cells isolated by Cook Medical gradient (40-80%) and followed up by swim-up techniques were used for real-time quantitative polymerase chain reaction (qPCR) and flow cytometry techniques to assess APN/CD13 and NEP/CD10 genes and proteins subsequently. Semen parameters were analyzed by computer-assisted sperm analysis. RESULTS: The findings revealed that there were significant differences in sperm total motility (41.07 ± 3.63 vs. 63.03 ± 3.31 %, P=0.0001), progressive motility (35.21 ± 2.64 vs. 20.93 ± 3.22%, P=0.001) and viability (69.9 ± 4.69 vs. 86.81 ± 1.26 %, P=0.002) in the addicted group vs. control ones. APN and NEP gene expression levels in the addicted group decreased compared with the control ones (1.00 ± 0.67 vs. 0.36 ± 0.13, P= 0.008 and 1.07 ± 0.11 vs. 0.52 ± 0.12 0.002, respectively). Flow cytometry analysis showed that the average percent of APN/CD13 in heroin consumers significantly decreased compared with the healthy ones, while NEP/CD10 rate between two groups was similar. We also observed that duration of drug dependence is correlated with sperm viability (r=-0.627, P=0.016) and motility (r=-0.410, P=0.05), NEP (r=-0.434, P= 0.049), and APN (r=-0.641, P=0.002) gene expression levels. CONCLUSION: We conclude that semen quality and enkephalin-degrading enzymes were altered in heroin-addicted men. other confirming the internal validity of our estimates.

The Effect of Heroin Addiction on Human Sperm Parameters, Histone-to-Protamine Transition, and Serum Sexual Hormones Levels.

PURPOSE: To investigate the effects of heroin on sperm parameters, histone-to-protamine transition ratios in mature sperm, and serum reproductive hormone levels in active heroin users. MATERIALS AND METHODS: Semen and blood samples were collected from 25 men who used only heroin for at least 12 months and the same number healthy men who did not use any drugs and did not suffer from infertility problems. Computer-based analysis, Aniline blue staining, and hormonal assessment were performed to provide valuable new information on the relationship between addiction and semen profile and serum reproductive hor-mone levels. RESULTS: Our finding showed that semen pH (7.8 vs. 7.75), sperm motility (42.93 ± 3.89% vs. 68.9 ± 2.68%), and viability (73.27 ± 3.85% vs. 86.48 ± 1.05%), and sperm histone replacement abnormalities (32.33 ± 10.89% vs. 5.56 ± 0.85%) were significant differences in addicted group vs. non-exposed ones (P ? .05). In addition, serum sex hormone levels were not significantly differed between groups. There was a correlation between the amount of daily heroin consumption and LH level. We also observed that duration of drug dependence is correlated with sperm abnormalities. CONCLUSION: We concluded that heroin consumption affect sperm maturities such as histone-to-protamine ratio and impair semen profile in general and particularly sperm morphology and motility. Heroin may be considered as one of the idiopathic male infertility reason.

Psychometric Properties of the Persian Adaptation of Mini-Cog Test in Iranian Older Adults.

The aim of this study was to assess the psychometric properties of the Mini-Cog in Iranian older adults. It was a cross-sectional study; 50 older people with dementia and 50 without dementia who matched for age, gender, and education entered the study. The diagnostic and statistical manual of mental disorders criteria for dementia were used as gold standard. A battery of scales included the abbreviated mental test score (AMTS), the Geriatric Depression Scale, and the Mini-Cog was performed. Validity and reliability of the Mini-Cog determined using the Pearson product-moment correlation coefficient (Pearson's r), Cronbach's alpha, and Receiver Operating Characteristic (ROC) curve analysis. The Persian version of Mini-Cog showed a good inter-rater reliability ( K = 0.76, p < .01) and a positive concurrent validity ( r = 0.39, p < .01) with the AMTS. The sensitivity and specificity were 88% and 62.8%, respectively, using the original cutoff point of 2. The findings showed that the Persian version of Mini-Cog have an acceptable sensitivity, specificity, and substantial overall agreement with the AMTS.

In vitro development of embryos from experimentally Kerack-addicted Mice.

BACKGROUND: Prenatal drug exposure, as a common public health concern, is associated with an increased risk of adverse effects on early embryo development. OBJECTIVE: To investigate the in vitro development of - embryo from experimentally Kerack-addicted mice. MATERIALS AND METHODS: Twenty-five female mice were studied in five groups: control, vehicle, and three experimental groups of Kerack-dependent mice (I, II, and III) which received different doses of Kerack for 14 days. After the establishment of addiction model (7 days), experimental groups I, II, and III were given Kerack intraperitoneally at the doses of 5, 35, and 70 mg/kg, twice a day for a period of 7 days, respectively. The vehicle group received normal saline and lemon juice whilst the control group just received water and food. Morulae were obtained through oviduct flashing. The survived embryos were cultured in T6+ 5mg/ml bovine serum albumin. The developmental rates up to hatched stage daily and embryo quality (differential staining and Tunnel staining) were also assessed. RESULTS: The developmental potential of embryos obtained from the addicted mother was significantly decreased in comparison with control group. There was a significant reduction in the rate of blastocyst formation in the high dose Kerack dependent group. However, in addicted mice there was reduction in the total cell number (40.92% vs. 65.08% in control) and, inner cell mass percentage (17.17% vs. 26.15% in control) while apoptotic cells numbers were increased (7.17 vs. 1.46 in control) (p<0.05). CONCLUSION: The Kerack addiction during pregnancy retards preimplantation development and induces apoptosis.

Having Multiple Sexual Partners among Iranian Intra-Venous Drug Users.

BACKGROUND: Transmission of HIV from intra-venous drug users (IDUs) to the community occurs predominantly through high-risk sexual behaviors. Limited information exists regarding the high-risk sexual behaviors of IDUs in Iran. AIM: The aim of this study was to determine the prevalence and factors associated with having multiple sexual partners among Iranian IDUs. METHODS: This is a national survey on drug-dependent adults. Participants were sampled from medical centers, prisons, and streets of capitals of 29 provinces in Iran between May 2007 and February 2008. We analyzed data of 1416 current IDUs. Socio-demographics and drug use characteristics were entered into a binary logistic regression model to determine predictors of having multiple sexual partners. RESULTS: Having multiple sexual partners in the past or at the time of survey was reported by 56.4% of Iranian IDUs. Multivariate analysis showed that the likelihood of having multiple sexual partners in IDUs decreased by being married [odds ratio (OR), 0.38; P < 0.001] and increased by female gender (OR, 13.44; P = 0.02), having illegal income (OR, 1.72; P = 0.003), higher monthly family income (OR, 1.01; P = 0.003), pleasure, curiosity, and recreation as cause of first drug use (OR, 1.37; P = 0.04), ruins as usual place for injection (OR, 1.89; P = 0.001) and history of syringe sharing (OR, 1.50; P = 0.02). CONCLUSION: Having multiple sexual partners was reported by majority of Iranian IDUs, and this was linked to socio-demographics, initiation data, and other risk behaviors. This information should be considered in prevention efforts to reduce sexual transmission of HIV infection in Iran.

The effects of poly L-lactic acid nanofiber scaffold on mouse spermatogonial stem cell culture.

INTRODUCTION: A 3D-nanofiber scaffold acts in a similar way to the extracellular matrix (ECM)/basement membrane that enhances the proliferation and self-renewal of stem cells. The goal of the present study was to investigate the effects of a poly L-lactic acid (PLLA) nanofiber scaffold on frozen-thawed neonate mouse spermatogonial stem cells (SSCs) and testis tissues. METHODS: The isolated spermatogonial cells were divided into six culture groups: (1) fresh spermatogonial cells, (2) fresh spermatogonial cells seeded onto PLLA, (3) frozen-thawed spermatogonial cells, (4) frozen-thawed spermatogonial cells seeded onto PLLA, (5) spermatogonial cells obtained from frozen-thawed testis tissue, and (6) spermatogonial cells obtained from frozen-thawed testis tissue seeded onto PLLA. Spermatogonial cells and testis fragments were cryopreserved and cultured for 3 weeks. Cluster assay was performed during the culture. The presence of spermatogonial cells in the culture was determined by a reverse transcriptase polymerase chain reaction for spermatogonial markers (Oct4, GFRα-1, PLZF, Mvh(VASA), Itgα6, and Itgß1), as well as the ultrastructural study of cell clusters and SSCs transplantation to a recipient azoospermic mouse. The significance of the data was analyzed using the repeated measures and analysis of variance. RESULTS: The findings indicated that the spermatogonial cells seeded on PLLA significantly increased in vitro spermatogonial cell cluster formations in comparison with the control groups (culture of SSCs not seeded on PLLA) (P≤0.001). The viability rate for the frozen cells after thawing was 63.00% ± 3.56%. This number decreased significantly (40.00% ± 0.82%) in spermatogonial cells obtained from the frozen-thawed testis tissue. Both groups, however, showed in vitro cluster formation. Although the expression of spermatogonial markers was maintained after 3 weeks of culture, there was a significant downregulation for some spermatogonial genes in the experimental groups compared with those of the control groups. Furthermore, transplantation assay and transmission electron microscopy studies suggested the presence of SSCs among the cultured cells. CONCLUSION: Although PLLA can increase the in vitro cluster formation of neonate fresh and frozen-thawed spermatogonial cells, it may also cause them to differentiate during cultivation. The study therefore has implications for SSCs proliferation and germ cell differentiation in vitro.

Needle and syringe sharing among Iranian drug injectors.

OBJECTIVE: The role of needle and syringe sharing behavior of injection drug users (IDUs) in spreading of blood-borne infections - specially HIV/AIDS - is well known. However, very little is known in this regard from Iran. The aim of our study was to determine the prevalence and associates of needle and syringe sharing among Iranian IDUs. METHODS: In a secondary analysis of a sample of drug dependents who were sampled from medical centers, prisons and streets of the capitals of 29 provinces in the Iran in 2007, 2091 male IDUs entered. Socio-demographic data, drug use data and high risk behaviors entered to a logistic regression to determine independent predictors of lifetime needle and syringe sharing. RESULTS: 749(35.8%) reported lifetime experience of needle and syringe sharing. The likelihood of lifetime needle and syringe sharing was increased by female gender, being jobless, having illegal income, drug use by family members, pleasure/enjoyment as causes of first injection, first injection in roofless and roofed public places, usual injection at groin, usual injection at scrotum, lifetime experience of nonfatal overdose, and history of arrest in past year and was decreased by being alone at most injections. CONCLUSION: However this data has been extracted from cross-sectional design and we can not conclude causation, some of the introduced variables with association with needle and syringe sharing may be used in HIV prevention programs which target reducing syringe sharing among IDUs.