Relationship between long non-coding RNA MALAT1 and HOTAIR expression with sperm parameters, DNA and malondialdehyde levels in male infertility.

BACKGROUND: Sperm quality is a complex index used to evaluate the fertility potential of men. The long non-coding RNA (lncRNA) MALAT1 participate in sperm development and HOTAIR have critical roles in the regulation of oxidative stress responses. This study aimed to evaluate the relationship of lncRNA MALAT1 and HOTAIR expression with sperm parameters, DNA fragmentation and malondialdehyde (MDA)levels in sperm fertility. METHODS: In this experimental study, semen samples (n = 30 fertile, n = 30 infertile) men were collected and evaluated for sperm parameters by computer-aided sperm analysis(CASA). Sperm DNA integrity quality was assessed by the Acridine orange(AO) test. MDA levels were determined by the Thiobarbituric acid reaction method. The expression of MALAT1 and HOTAIR was detected by RT-PCR. RESULTS: We observed a decreased level of MALAT1and HOTAIR expression in the infertile men (p < 0.001). The relative expression level of MALAT1and HOTAIR showed a positive correlation with motility and morphology (p < 0.001). Subsequently, we found the DNA damage and MDA levels was negatively correlated with expression level of genes of sperm (p < 0.001). CONCLUSION: In this study the low expression of MALATI and HOTAIR resulted in the high level of MDA, DNA damage, and reduced motility of sperm. This study suggests the therapeutic opportunities in respect to MALATI and HOTAIR expression in the sperm function.

Correlation of Obesity and Serum Vitamin D Levels with Sperm DNA Integrity, Sperm Quality, and Sperm Viability in Normozoospermia Men.

Background: Obesity, Vitamin D (VD) deficiency, and infertility are important ubiquitous issue; however, the association of obesity and serum VD levels with abnormal sperm is unclear and inconclusive. The current study investigated the correlation of obesity and serum VD levels with sperm DNA integrity and sperm parameters in normozoospermia men. Materials and Methods: Semen and blood samples from 64 men were divided into two groups: obese and nonobese men based on body mass index (BMI). Sperm motility and viability were determined by computer-aided sperm analysis and eosin-nigrosin staining. DNA fragmentation, determined by sperm chromatin dispersion method. VD concentrations were assessed by the Elisa technique. Results: Serum concentration of VD levels in the obese group was significantly lower than nonobese men (P < 0.05). Sperm motility was significantly reduced in the obese group in comparison to nonobese (P < 0.05). Rapid progressive motility was statistically lower in obese men compared with the nonobese group (P < 0.05). Sperm count and morphology were not statistically significant in both groups. Sperm viability in the nonobese group was significantly decreased in comparison to obese group (P < 0.05). DNA integrity was significantly higher in the obese group as compared with nonobese (P < 0.01). Conclusion: VD deficiency in the obese group showed decreased sperm motility, increased DNA damage, and viability. Adverse consequences of obesity and the possible effect of BMI infertility treatment must be discussed with counseling couples interested in assisted reproductive techniques outcomes, especially in men without any unknown cause.

Human thumb consists of three phalanges and lacks metacarpal? A morphometric study on the long bones of the hand.

PURPOSE: For many years, it was thought that the thumb consists of just two phalanges that differentiate it from the other four medial triphalangeal fingers. But there are some old reports that few former scientists believed the thumb has three phalanges and it lacked a metacarpal, and the thumb metacarpal is a phalanx. So this anthropometric study was carried out by investigating the morphology of the long bones of the hand and correlations between the thumb metacarpal and other miniature long bones of the hand. METHODS: We studied anterior-posterior X-ray images of the right hands of 80 individuals from 18 to 65 years old. The exploration targets were the length of all metacarpals (MC), proximal phalanges (PP), middle phalanges (MP), and distal phalanges (DP). Friedman Repeated Measures Analysis of Variance and Dunn's post hoc test were carried out to compare the means of all variables. The correlation between all quantitative factors was done by Spearman Rank Correlation (Spearman's Rho) coefficient. RESULTS: Our results showed that the length of the phalanges and the total length of the fingers are independent of the related metacarpal length (P < 0.001). Also, the thumb metacarpal length in comparison to all bones of the hand was significantly different from all long bones of the hand except the proximal phalanx of the middle finger (P = 1). CONCLUSION: Based on the morphology of the long bones of the hand and the high similarity between the thumb metacarpal and phalanges especially the proximal phalanx of the middle finger, it can be suggested that the current thumb metacarpal is a proximal phalanx of the thumb.

Correlation of Treatment of Peptic Ulcer Bleeding by Argon Plasma Coagulation (APC) via Contact Heat Probe Method (heater probe) with Epinephrine Injection.

BACKGROUND This clinical investigation aimed to compare the efficacy of treatment of peptic ulcer hemorrhage by argon plasma coagulation (APC) via contact heat probe method (heater probe) along with epinephrine injection. METHODS 100 patients who underwent endoscopic treatment, were randomly divided into two groups consisting of 50 patients each. In the first group, an intervention was performed using foot pedal and 2.3 mm and 3.2 mm argon probes placed in a 2 to 8 mm distance of delivery place leading to plasma coagulation, sufficient necrosis and hemostasis. In the second group, wound press contact probe was used for wound healing with 15 watts of heat for about 25 degrees, causing coagulation and hemostasis. To evaluate and compare the ulcer treatment in both groups, the patient progress results were monitored for a period of one month from the day of discharge. Statistical analyses of data were performed using SPSS software version 22 along with Chi-square test and T-test. RESULTS No significant difference observed in two groups in term of age, sex and clinical symptoms, but patients treated with APC method had higher hemoglobin levels (p < 0.001). The duration of intervention and abdominal bloating in APC group was significantly higher with two cases of re-admission. In HP group, 3 cases (6.3%) had treatment failure and an average transfused blood was significantly higher in the HP group (p < 0.001). CONCLUSION Endoscopy treatment duration was significantly lower in patients treated with the HP method due to separate washing route. HP method seems to be more appropriate for treatment of cases with abdominal bloating, distal gastric lesion and HP bulbs.

Effects of temperature and storage time on the motility, viability, DNA integrity and apoptosis of processed human spermatozoa.

The aim of this study was to evaluate motility, viability, DNA integrity and apoptosis of spermatozoa when washed semen samples were kept for up to 12 days at 4-6°C and 25°C. In this experimental study, 26 normozoospermic semen samples were washed twice in Modified Ham's F10 and resuspended in IVF fertilisation medium. Half of the specimens were stored at 4-6°C, and the other half was kept at 25°C for 12 days. The proportions of viable, motile, spermatozoa with double-stranded DNA and apoptotic spermatozoa were examined during storage time. Apoptosis was measured using annexin V-PI staining followed by flow cytometry. Results showed that sperm motility and viability decreased during 12 days of sample storage (p < .001). There was no significant difference between the two temperatures in terms of motility and viability for up to 2 days (p < .05). The percentage of spermatozoa with double-stranded DNA remained unchanged during the 12 days of storage at both temperatures (p > .05). Although there was no difference between the two temperatures in terms of motility, viability and apoptosis during the first two days of storage, storage of spermatozoa at 4-6°C is better than storage for a longer period than storage at 25°C. Sperm DNA resisted against denaturation during storage.

Correlation between sperm motility and sperm chromatin/DNA damage before and after cryopreservation and the effect of folic acid and nicotinic acid on post-thaw sperm quality in normozoospermic men.

Cryopreservation exposes sperm to physical and chemical stresses causing cell damages and impairs sperm functions. The aim of this study was to evaluate the association between motility and sperm chromatin/DNA damage before and after cryopreservation and investigate the effects of folic acid and nicotinic acid on post-thaw sperm quality. Thirty semen samples were obtained from 30 normozoospermic men, aged between 25 and 45 years old. Each sample were divided into five aliquots to form the following groups: fresh, cryopreserved with sperm-freeze only (control), with nicotinic acid (10 mM), with folic acid (50 nM), and with a combination of folic acid (50 nM) + nicotinic acid (10 mM). Sperm viability and motility in each group were assessed by eosin-nigrosine staining and computer-aided sperm analysis respectively. Sperm chromatin quality was studied by aniline blue, toluidine blue, acridine orange staining methods and sperm chromatin dispersion test. Cryopreservation led to a significant reduction in sperm quality in comparison to fresh sample groups (p < 0.05). Sperm chromatin damage was negatively correlated with the percentage of progressively motile cells. Supplementation of the cryopreservation medium with folic acid or nicotinic acid induced a significant improvement in sperm parameters and chromatin quality, compared to control groups (p < 0.05). Meanwhile, the combination of folic acid + nicotinic acid showed a significant protective effect in post thaw sperm. In conclusion, cryopreservation generated oxidative stress, inducingsperm cryodamage, reducing progressive motility and sperm quality, as an indicator of significant chromatin/DNA damage. Folic acid and nicotinic acid exhibited a potential cryoprotective effect by enhancing sperm quality.

Effect of preservation of human semen sample at 4-6 and 25 °C on sperm motility.

Sperm motility is the result of transverse movements that exist along its tail. It plays an important role in male fertility. The aim of this study was to evaluate the influence of keeping washed normozoospermic semen samples at 4-6 and 25 °C on the motility of spermatozoa. 26 semen samples of normozoospermic were washed twice in modified Ham's F10 medium. Then, thirteen of the semen samples were kept in refrigerator (4-6 °C) and the remaining samples were stored in incubator (25 °C) for 12 days. On the 0 (immediately after sampling as control group), 1st, 2nd, 5th, 7th and the 12th days, the percentage of fast progressive (grade a), slow progressive (grade b), non-progressive (grade c) and immotile (grade d) sperm cells were calculated for each temperature. The data obtained from this study showed that the percentages of a, b and c grades of motile spermatozoa were significantly decreased (p < 0.001) during 12 days at the both temperatures but reduction of these percentages has a gentle slope at 4-6 °C. There was no motile sperm after 12 days of storage. This study suggests that motile spermatozoa could be retrieved up to 7 days after the storage of washed normozoospermic men semen samples at 4-6 and 25 °C. Also, there were no motile sperm cells 12 days after sampling.

The effects of vitamin C and menthone on acyclovir induced DNA damage in rat spermatozoa: An experimental study.

BACKGROUND: Acyclovir (ACV) is known to be toxic to gonads, inducing apoptosis in the reproductive system. The beneficial effects of vitamin C (Vit C) and menthone, both as antioxidant agents on various organs has been reported. OBJECTIVE: This study evaluated the potential role of the Vit C and menthone on the DNA damage in rat spermatozoa induced by the ACV. MATERIALS AND METHODS: In this experimental study, adult male albino Wistar rats with average weight of 250±10 gr, were divided into six groups (n=18/each), as: ACV (15 mg/kg/day), ACV+Vit C (20 mg/kg/day), ACV+ menthone (100 µl/d), ACV+ menthone (250 µl/d), ACV+ menthone (400 µl/day) and control group without any treatment. At the end of experiment, the animals were sacrificed and sperm samples were collected and isolated in phosphate-buffered saline and examined by TUNEL staining process. The percentage of TUNEL positive spermatozoa was evaluated by fluorescence microscopy. Each experiment was performed in three repeats. RESULTS: Male rats exposed to ACV had significant increase in DNA damages in comparison to other groups. The percentage of TUNEL positive sperm cells was 90.83 (p<0.001) in ACV group. The protective role of both antioxidants used in high dose, compensate the adverse effects of the ACV. The results showed that the percentage of apoptotic sperm in the ACV+Vit C group was 16.38 (p<0.001) and in the ACV+ menthone (400 µl/d) group was 16.05 (p<0.001). CONCLUSION: The present results showed that Vit C and menthone at higher dose have a good compensatory effect with significant reduction in DNA damages in sperm cells by reversing the adverse effect of ACV on the reproductive system in male rat.

Thymoquinone as a natural spermostatic substance in reproductive medicine: An experimental study.

BACKGROUND: Nonoxynol-9 a nonionic surfactant is widely used for its spermicidal effects. Finding new sperm immobilizing agents is necessary because Nonoxynol-9 damages the tissues of female reproductive system. OBJECTIVE: The aim of this study was to evaluate the effects of Thymoquinone (TQ) as a potential spermostatic compound on the motility and viability of human spermatozoa. MATERIALS AND METHODS: In this experimental study, the effects of 5, 10, 20, 50, 100 µg/ml, 1 and 10 mg/ml of TQ on normozoospermic semen samples were investigated. Sperm motility and viability were compared between untreated and TQ-treated aliquots of each semen sample. To evaluate the effects of TQ on the alteration of mitochondrial membrane potential (MMP), 32 semen samples were examined using 50 µg/ml of TQ. Flow cytometric analysis was performed after staining of spermatozoa with JC-1. RESULTS: Doses above 20 µg/ml of TQ could eventually immobilize all spermatozoa in culture medium. Adding 50 µg/ml of TQ did not significantly diminish the percentage of viable spermatozoa and flow cytometry results revealed that this amount of TQ could decrease sperm MMP. CONCLUSION: TQ could discontinue the movement of sperm cells in medium without reducing the population of live spermatozoa. It is more likely that TQ exerts its spermostatic action by mitigating the MMP of spermatozoa. Therefore, TQ could be considered as a potential new natural spermostatic chemical.

The effect of vitamin E on neuronal apoptosis in hippocampal dentate gyrus in rabbits fed with high-cholesterol diets.

BACKGROUND: Hypercholesterolemia that can increase stress oxidative has destructive effects on brain functions. Vitamin E is a powerful antioxidant and its effects on decrement of oxidative stress in the diseases such as Alzheimer's and hypercholesterolemia are demonstrated. The aim of this study was evaluation of the effects of vitamin E on the level of neuronal apoptosis in granular layer of dentate gyrus in the rabbits that fed with high-cholesterol diet. MATERIALS AND METHODS: Male New Zealand white rabbits were divided into the control, the Vitamin E (50 mg/kg; gavage), the high-cholesterol diet (containing 2% cholesterol), and the high-cholesterol diet-vitamin E groups. Serum levels of cholesterol, LDL, and HDL, before and after the regimen for 6 weeks, were measured. Then, the rabbits for immunohistochemical staining (TUNEL Test) and evaluation of neuronal apoptosis in dentate gyrus of hippocampal formation were anesthetized and brains were dissected. RESULTS: Results showed that after the regimens, serum levels of cholesterol, LDL, and HDL in the cholesterol receiving groups were increased significantly (P < 0.05). Histological results demonstrated that neuronal apoptosis in the dentate gyrus of the high-cholesterol diet group was increased significantly (P < 0.05) comparing to the control group; however, vitamin E decreased apoptosis as there wasn't any significant differences between the high-cholesterol diet-vitamin E and control groups. CONCLUSIONS: Present results showed that consumption of high-cholesterol diets through hypercholesterolemia and its complication can induce neuronal death in hippocampus and probable resulting cognition disorders; however, vitamin E has neuroprotective effects and prevents neuronal apoptosis significantly.

The effect of aqueous cinnamon extract on the apoptotic process in acute myeloid leukemia HL-60 cells.

BACKGROUND: Acute promyelocytic leukemia (APL) is an acute leukemia diagnosed by translocation of chromosomes 15 and 17 [T (15,17)] and aggregation of neoplastic promyelocytes which are incapable of being converted into mature cells. Today, many tend to use medicinal herbs in studies and clinical applications for treatment of cancers. Cinnamon with scientific name "cinnamomumzelanicum" is a shrub of Laurales order, lauraceae family with cinnamomum genus. It is a medicinal shrub with anti-proliferation effect on tumor cells. This study was conducted to determine the effects of aqueous cinnamon extract on HL-60 cells as a model for APL. MATERIALS AND METHODS: In this in vitro experimental study, HL-60 cell line was cultured under the influence of cinnamon extract's concentrations of 0.01, 0.1, 1, and 2 mg/ml in with intervals of 24, 48, and 72 h. Growth inhibition and toxic effects of cinnamon extract were evaluated through tetrazolium salt reduction. The effect of this herb on the cell cycle was studied by flow cytometry. The Hoechst stain was used to detect apoptotic cell nuclei. RESULTS: Cinnamon extract inhibited the growth of HL-60 cells as correlated with concentration and time. After 72 h of treating HL-60 cells with 0.01 mg/l cinnamon extract, the growth of cells was inhibited by 90.1%. Cinnamon extract stopped the cell cycle in G1 phase and the Hoechst staining verified the apoptotic process in those cells. CONCLUSION: Considering the inhibitory property of cinnamon extract, we recommend it as a single drug or besides other medications for treating promyelocytic leukemia.

Evaluation of TERT mRNA alternative spliced variants in aortic endothelial cells of the rabbit, in the fatty streaks stage of atherosclerosis.

OBJECTIVES: There are some lines of evidence about the role of telomeres in atherosclerotic plaque formation and development. Despite the role of telomerase in telomere elongation and protection, its role in atherosclerosis is not fully understood. The present study was designed to evaluate the importance of alternative splicing variants ofTERT (telomerase reverse transcriptase) in the atherosclerosis process. METHODS AND RESULTS: Six Dutch-Polish rabbits were fed a high-cholesterol diet, and six control animals a normal diet. Alternative splicing was assessed by RT-PCR in aortic endothelial cells of these rabbits. Fatty streaks formation was assessed by H & E staining of aortic tissues. There were no significant differences in variants of TERT in control and test animals; neither in endothelial cells nor in aortic vessel wall. CONCLUSION: Our results demonstrated that a decrease in telomerase activity in atherosclerosis formation is not due to TERT gene alternative splicing.

Triiodide PVC membrane electrodes based on charge-transfer complexes.

Three new electrodes were prepared by incorporating two different charge-transfer complexes and amino crown ether into plasticized PVC membranes. The electrodes showed Nernstian response to triiodide ion over the activity range from 1.0 x 10(-5) to 1.0 x 10(-1) mol x L(-1) with detection limits at approximately 1.0 x 10(-6) mol x L(-1). The resulting electrodes have fast response times (20-30 s) and good stabilities (4 months) and can be used over a wide pH range of 2.5-9.0. The proposed electrodes exhibit anti-Hofmeister behavior with excellent selectivity toward triiodide ion against a wide range of common interferences. Comparative study suggests that amino (aza) crown ether alone or in the form of a charge-transfer complex with iodine, as an ionophore in a PVC liquid membrane, is sensitive to triiodide ion. The electrodes were used as indicator electrodes in potentiometric titration of triiodide ion against thiosulfate ion.