Evaluation of L-Carnitine Potential in Improvement of Male Fertility.

L-carnitine, through its antioxidant potential, plays a significant role in reducing ROS production in male genital tract; therefore, fundamental improvements in spermatogenesis process and sperm structural and functional parameters in seminal plasma can be observed by treatment with L-carnitine. A literature search was performed using PubMed (including Medline) from the database earliest inception to 2021. Eligibility criteria included studies on protective effects of L-carnitine against damages to the male reproductive system. Based on the findings of the current study, L-carnitine has an effective potential to protect testis and improve conventional and functional sperm parameters against ROS-induced damages by sperm cryopreservation, busulfan treatment, and radiation.

Effect of Pre-Incubation of Cryopreserved Sperm with either Kisspeptin or Glutathione to Mitigate Freeze-Thaw Damage.

Background: Sperm cryopreservation reduces sperm quality. Kisspeptin (KP) has beneficial effects on sperm functions. This study compares the effect of KP and Glutathione (GSH) on mitigating the detrimental effects of the freeze-thaw cycle on sperm. Methods: An experimental study was conducted in Birjand (Iran) during 2018-2020. Thirty normal swim-up semen samples were treated with Ham's F10 medium (negative control), 1 mM GSH (positive control), or KP (10 µM) for 30 min before freezing. The motility, acrosome reaction, capacitation, and DNA quality of the frozen-thawed sperms were assessed according to the WHO guidelines. Statistical analysis was performed using paired t test, one-way analysis of variance, and least significant difference. Results: Pre-incubation with KP significantly increased the percentage of sperm motility (34.00±6.7, P=0.003) compared to the control (20.44±7.4) and GSH-treated (31.25±12.2) aliquots. The frequency of non-capacitated spermatozoa was significantly higher in the KP-treated group (98.73%) than in the control (96.46%) and GSH-treated (96.49%) aliquots (P<0.001). The percentage of acrosome-intact spermatozoa in the KP-treated group (77.44%) was significantly higher than the control (74.3%) and GSH-treated (74.54%) groups (P<0.001). The sperm frequency with normal histone in the KP-treated group (51.86%) and with normal protamine (65.39%) was significantly higher than the controls (P=0.001 and P=0.002, respectively). The percentage of TUNEL-positive sperm was significantly lower in the KP-treated group (9.09±2.71) than both GSH-treated (11.22±2.73) and control (11.31±2.2) groups (both P=0.002). Conclusion: Pre-incubation with KP protects sperm motility and DNA integrity from the detrimental effect of the freeze-thaw cycle. KP is suitable as a pre-treatment to control sperm quality during freezing-thawing.

Resveratrol ameliorates bisphenol A-induced testicular toxicity in adult male rats: a stereological and functional study.

BACKGROUND: Bisphenol A (BPA) is one of the most widely used synthetic chemicals worldwide. BPA as an endocrine disruptor affects the reproductive systems through estrogenic and antiandrogenic proprieties. Resveratrol (RES) as a natural polyphenol and potent antioxidant exhibits protective effects against reproductive toxicity by inhibiting of oxidative stress. 48 male rats were divided into eight groups (n=6), including CONTROL, OLIVE OIL (0.5 ml/ day), Carboxy methylcellulose (CMC) (1 ml of 10 g/l), RES (100mg/kg/day), low dose of BPA (25 mg/kg/day), high dose of BPA (50 mg/kg/day), low dose of BPA + RES, and high dose of BPA + RES. All treatments were done orally per day for 56 days. At the end of the 8th week, blood samples were collected for hormone assays. Then, the sperm parameters were analyzed, and the left testis was removed for stereological study. RESULTS: We showed a significant decrease in sperm parameters in the low and high doses of BPA groups compared to control groups (P<0.05). The volume of testicular components as well as the diameter and length of seminiferous tubules significantly reduced (11-64 %), and the total number of the testicular cell types decreased (34-67 %) on average in the low and high doses of BPA groups. Moreover, serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone hormones concentration showed a significant reduction in both doses of BPA groups (P<0.01). Nonetheless, treatment with RES could ameliorate all the above-mentioned changes in the low and high doses of BPA groups (P<0.05). CONCLUSIONS: RES could prevent BPA-induced testicular structural changes and sperm quality via improving gonadotropin hormones and testosterone levels.

RèSUMè: CONTEXTE: Le bisphénol A (BPA) est l'un des produits chimiques synthétiques les plus utilisés dans le monde. Le BPA en tant que perturbateur endocrinien affecte le système reproducteur par le biais de ses propriétés œstrogéniques et anti-androgènes. Le resvératrol (RES), en tant que polyphénol naturel et puissant antioxydant, présente des effets protecteurs contre la toxicité sur la reproduction en inhibant le stress oxydatif. Quarante-huit rats mâles ont été divisés en huit groupes (n = 6), comprenant les groupes TÉMOIN, HUILE D'OLIVE (0,5 ml/jour), méthylcellulose Carboxyle (MCC) (1 ml de 10 g/L), RES (100 mg/kg/ jour), faible dose de 25 de BPA (25 mg/kg/jour), dose élevée de BPA (50 mg/kg/jour), faible dose de BPA + RES et dose élevée de BPA + RES. Tous les traitements ont été effectués quotidiennement par voie orale pendant 56 jours. À la fin de la 8ème semaine, des échantillons de sang ont été prélevés pour dosages hormonaux. Ensuite, les paramètres du sperme ont été analysés et le testicule gauche a été retiré pour une étude stéréologique. RéSULTATS: Nous avons montré une diminution significative des paramètres spermatiques dans les groupes traités par doses faibles et doses élevées de BPA par rapport aux groupe témoin (P<0,05). Le volume des composants testiculaires ainsi que le diamètre et la longueur des tubules séminifères ont été considérablement réduits (11-64 %) ; le nombre total des types de cellules testiculaires a diminué (34-67 %) en moyenne dans les groupes traités par doses faibles et doses élevées de BPA. De plus, la concentration sérique d'hormone folliculostimulante (FSH), lutéinisante (LH) et de testostérone a montré une réduction significative dans les groupes traités quelle que soit la dose de BPA (P<0,01). Néanmoins, le traitement par RES pourrait améliorer tous les changements mentionnés ci-dessus dans les groupes traités par doses faibles et élevées de BPA (P<0,05). CONCLUSIONS: Le RES pourrait avoir un effet positif sur les changements structurels testiculaires induits par le BPA, ainsi que la qualité du sperme, en améliorant les taux sériques d'hormones gonadotrophines et de testostérone. MOTS-CLéS: Bisphénol A Resvératrol Toxicité testiculaire Paramètres du Sperme Stéréologie.

L-Carnitine and Pentoxifylline Supplementation Improves Sperm Viability and Motility at Low Temperature.

BACKGROUND: In infertility clinics, preserving high-quality spermatozoa for a long time is a necessity. Pentoxifylline (PT) and L-carnitine (LC) are effective in improving sperm motility as well as protecting the sperm membrane. The present study aimed to investigate the protective impacts of PT and LC on the quality of the normal sperm motility, protamine content, and viability on prolonged storage for 12 days at 4-6°C. MATERIALS AND METHODS: The present experimental work included 26 samples, which were first prepared based on the swim-up technique, of normozoospermic men. They were divided into three aliquots as untreated control, LC-treated, and PT-treated groups and incubated for up to 12 days at 4-6°C. Thereafter, chromatin maturity, sperm viability, and motility were assessed on 0, 1, 2, 5, 7, and 12 days. Data were analyzed using a one-way analysis of variance. RESULTS: The obtained data revealed that PT supplementation increased the percentage of motile spermatozoa in comparison with control and LC-treated specimens. On the other hand, LC supplementation increased the percentage of viable spermatozoa in comparison with the PT-treated and control samples. During the 12-day storage, the percentage of spermatozoa with a normal protamine content was nearly unchanged in the three groups (P>0.05). CONCLUSION: Although LC supplementation can be considered a better alternative than PT for preserving sperm viability, PT could better preserve sperm motility compared to LC during 12 days at 4-6°C.

Effects of kisspeptin incubation on the mature mouse testicular sperms quality: An experimental study.

Background: Sperm quality has an important role in the success of assisted reproductive techniques, by adding some bioactive agents with a positive impact on sperms, it can be improved. Objective: This study aimed to evaluate the effects of kisspeptin on the sperm motility criteria, Lactate dehydrogenase-C (LDHC) activity, acrosomal reaction, and capacitation in the mouse testicular sperm in vitro. Materials and Methods: Sperm samples were extracted from testes of 96 male Balb/C mice weighing 25-30 gr, aged 6-8 wk. Then, they were separated into 4 parts; 2 controls and 2 kisspeptin-treated aliquots; each one incubated for either 15 or 30 min. The sperm motility and the LDHC activity were evaluated, and also the frequency of the non-capacitated, intact, and acrosomal-reacted sperms were evaluated by staining with Wheat germ agglutinin, Peanut agglutinin, and Concanavalin A, respectively. The stained sperms were analyzed by flow cytometry and fluorescent microscope. Results: Our result showed that kisspeptin increased both the sperm motility (p = 0.04) and LDHC enzyme activity (p = 0.04) after 15 min of incubation. At the same time, it did not impact the frequency of the non-capacitated, intact and acrosomal-reacted sperms after incubation in the same period (p = 0.16). Conclusion: A 15 min period of incubation with kisspeptin could be applicable for evaluating sperm motility and LDH activity.

Histochemical Study of the Rat Uterine Glycoconjugate Alteration following Treatment with Exogenous Gonadotropic Hormones during the Implantation Period.

One of the female causes of infertility is anovulation which is treatable with gonadotropin hormones. These hormones affect the molecular organization of the uterus such as glycoconjugates that are the first site of contact between the blastocyst and the uterus. The objective of this project was to study the alteration of glycoconjugates on the uterine apical, Golgi zone, and basement membrane of epithelial cells and the uterine gland after hyperstimulation with pregnant mare serum gonadotropin (PMSG) (4, 8, 16, 24, and 40 IU), during the implantation period. Injection of PMSG (in experimental groups) and injection of distilled water (in the control group) were followed by HCG administration (10 IU), mating, isolation of positive vaginal plug rats, and killing at 5.5 days of pregnancy. Histochemistry was done on the pregnant uterine horns with the use of WGA, DBA, PNA, ConA, SBA, and UEA lectins. The intensity of the immunohistochemical staining was scored, and quantitative data were generated. 4 IU did not show any significant differences with the control, 8 IU had less effect on the alteration of the Golgi zone, and apical and basement membrane glycoconjugates and 40 IU had the least effects on the alteration of uterine gland glycoconjugates. Also, 24 IU had the most effect on the alteration of uterine glycoconjugates. Understanding of the effects of gonadotropin hormones at the uterine level in implantation time helps to optimize hormonal manipulation for improving the outcome of assisted reproductive procedures. It seems that the optimal dose for superovulation and less alteration in uterine glycoconjugates of rats at implantation time were induced by the administration of 8 IU PMSG.

Lectin Profile Variation in Mesenchymal Stem Cells Derived from Different Sources.

Human mesenchymal stem cells (MSCs), a promising source of stem cells for regenerative medicine, have different morphological and functional characteristics. Carbohydrate moieties on the cell surface play an important role, including cell-cell interaction and cell recognition. The objective of this study was to determine possible differences in glycoconjugate distribution patterns of MSCs derived from various sources. MSCs were isolated from adipose tissue, bone marrow, Wharton's jelly, and cord blood. Then, they were stained with FITC-conjugated wheat germ agglutinin (WGA), peanut agglutinin (PNA), concanavalin A (ConA), Ulex europaeus (UEA), Dolichos biflorus (DBA), and Atto-488 conjugated Phytolacca americana (PWM) lectins. The intensity of the reactions was scored using ImageJ software. Flow cytometry was performed to detect the expression of the endothelial marker CD144. The obtained data were analyzed by ANOVA and LSD. Cord blood-derived MSCs showed the most significant staining intensities with all lectins. All MSCs were also moderately stained with PNA. Bone marrow-derived MSCs failed to react with UEA, DBA, and ConA. Wharton's jelly-derived MSCs could also not be stained with ConA. Cord blood-derived MSCs contained 2 subpopulations: osteoclast- and fibroblast-like cells. Both lectin staining intensity and distribution pattern were different in these 2 cell types; therefore, the central part of osteoclast-like cells stained more intensive with PNA and PWM, while that part in fibroblast-like cells stained more intensive with ConA. None of them expressed CD144. The glycoconjugate content of MSCs derived from various sources is different.

Effects of pentoxifylline on the histological and ultra-structural features of vitrified mouse ovarian tissue: An experimental study.

BACKGROUND: Vitrification is a process that can be used to preserve gonads in the healthy and natural status. Oxidative stress is one of the disadvantages of vitrification. Pentoxifylline (PTX) is an antioxidant that can reduce reactive oxidative stress effects. OBJECTIVE: We aimed to investigate the effects of PTX on histological and ultra-structural features of vitrified and non-vitrified mouse ovarian tissue. MATERIALS AND METHODS: Twenty-five adult female Balb-C mice were randomly and equally divided into control group: the ovaries did not receive any treatment; experimental 1 and 2: the vitrified ovaries were incubated in phosphate buffer solution and bovine serum albumin without and with PTX, respectively, for 30 min; sham 1 and 2: the non-vitrified ovaries were incubated in phosphate buffer solution and bovine serum albumin and were incubated without and with PTX, respectively for 30 min. The right and left ovaries in all of the groups were evaluated using light and transmission electron microscopy, respectively. RESULTS: The histological and ultra-structural features of vitrified ovaries were seriously damaged. There was non-uniformed germinal epithelium and tunica albuginea, degenerated granulosa cells and stromal cells, puffy basement membrane and irregular thickness of zona pellucida, as well as a pyknotic nucleus and bubbly and segmented ooplasmic in the follicles. Also, ovarian tissues were damaged by the PTX in the non-vitrified ovaries. CONCLUSION: Vitrification can damage the histological and ultra-structural features of the ovary in mouse models. PTX as an antioxidant, with concentration of 1.8 mM could not prevent and restore these damages and had no adequate effects on the vitrified ovarian tissues.

Protective effects of curcumin co-treatment in rats with establishing chronic variable stress on testis and reproductive hormones.

BACKGROUND: Protracted and repeated exposure to chronic variable stress (CVS) may lead to reproductive dysfunction. It is a basic cause of male infertility. Curcumin (CUR) is an active fraction of turmeric that used in traditional Chinese medicine. CUR represents various pharmacological activities. OBJECTIVE: The purpose of this study was to determining the effects of CUR on testis and testosterone, follicle stimulating hormone (FSH) and luteinizing hormone (LH) in rats with establishing chronic variable stress. MATERIALS AND METHODS: Twenty-one adult male Sprague-Dawley rats were divided into three groups: 1) control, 2) CVS and 3) CVS+ CUR (100 mg/kg/day dissolved in 0.5 mL of olive oil). All of the animals in control, CVS, and CVS+CUR groups were sacrificed after 15 days. Testosterone, FSH, LH, and testis damage were evaluated. RESULTS: Significant changes in the normal range of testosterone, FSH, LH serum levels and seminiferous tubule apoptotic cells were detected in CVS group compared to the control rats (p=0.02). These parameters changed to a less extent in CVS+CUR animals compared to the CVS rats (p=0.02). CONCLUSION: Our findings propose that curcumin might have curative potential on the reproductive system function and its impairment. It's regulated by stress and reproductive-related hormones.

Kisspeptin expression features in the arcuate and anteroventral periventricular nuclei of hypothalamus of letrozole-induced polycystic ovarian syndrome in rats.

PURPOSE: Polycystic ovary syndrome (PCOS) is the most common endocrine disorder of the reproductive system characterized by polycystic ovaries and androgen excess. Letrozole is a nonsteroidal aromatase inhibitor that is used in experimental research to induce PCOS. Kisspeptin is an essential protein in regulation of cyclicity. Kisspeptin receptor is expressed in the hypothalamus and pituitary glands, and kisspeptin containing neurons are affected from sex steroid hormones. We aimed to investigate the number of kisspeptin-positive cells in the arcuate (Arc) and anteroventral periventricular nuclei (AVPV) of hypothalamus in the letrozole-induced PCOS. METHODS: 40 female Wistar rats were divided into the proestrus control, diestrus control, proestrus vehicle, diestrus vehicle and letrozole. Animals were sacrificed after 3 weeks, and sera, ovary and brain samples were harvested for further evaluations. RESULTS: Letrozole group had high weight gain, high numbers of ovarian follicular cysts, high levels of luteinizing hormone and testosterone and increase number of kisspeptin-positive cells in the Arc nucleus, as compared with the control groups (P ≤ 0.05 vs. proestrus control and proestrus vehicle). Letrozole group showed a decrease in the number of kisspeptin-positive cells in the AVPV nucleus (P ≤ 0.05 vs. proestrus control and proestrus vehicle). CONCLUSION: Our findings show that the number of kisspeptin-positive cells may be affected from letrozole, and that the changes in the number of these cells may be in favor of the appearance of PCOS features in this group.

Stress affects surface glycoconjugates of the rat endometrium at the time of implantation.

One of the treatments to infertility is In Vitro Fertilization (IVF). In the course of IVF, fertilization rate can be improved through stress reduction. Probably one of the causes of low outcome of IVF is changes in uterine glycoconjugates that are first site of contact between blastocyst and uterus, due to stress, e.g., stress of injection. Thus, the study of the injectional stress effects on implantation period is very important to improve the outcome of IVF. Sixteen mature female rats were divided to experimental and control groups. Experimental rats injected with 0.5cm3 distilled water intraperitoneally in diestrus or proestrus and 10 I.U HCG in estrus phase. Control rats injected only with 10 I.U HCG in estrus phase. The experimental and control rats mated with proven fertile male rats, sacrificed at 5.5 day of gestation (time of implantation) and their uterus horns removed. Uterine sections were stained with WGA, DBA, PNA, ConA, SBA and UEA lectins and grading of the intensity of the reaction in apical membrane, Golgi zone and basement membrane of uterine epithelial cells and uterine glands were performed by an arbitrary method. The intensity of the reaction to WGA and DBA in apical membrane and Golgi zone was significantly high in experimental group. It seems that injectional stress can decrease the rate of implantation through alteration in uterine glycoconjugates, e.g. increase in negatively charged glycoconjugates such as sialic acid, which reduce the receptivity of uterus for blastocyst.

Cystoisosporiasis-related human acalculous cholecystitis: the need for increased awareness.

Cholecystitis is one of the common surgical indications affecting human beings in many countries. A variety of infectious agents can be associated with acute or chronic acalculous cholecystitis, especially in HIV/AIDS patients. In this investigation, the authors aim to describe two cases of histologically and molecularly documented cystoisosporiasis (syn. isosporiasis) as the cause of chronic acalculous cholecystitis in two immunodeficient patients. During microscopic examinations of more than 2500 diarrheic patients' samples, 11 cases of cystoisosporiasis-related recurrent persistent/chronic diarrhea were detected. A review of the medical records of Cystoisospora belli (syn. Isospora belli)-positive patients showed that two of them, i.e. a patient with prolonged corticosteroid therapy and an AIDS patient, several months prior to fecal examinations had undergone cholecystectomy due to acalculous cholecystitis. The study was continued by a review of the histopathological investigation of the recuts prepared from the excised gallbladder tissue sections and stained with hematoxylin and eosin in order to detect a possible specific clinical correlation with cystoisosporiasis. Light microscopic examination revealed the presence of various developmental stages of a coccidial parasite, namely Cystoisospora belli, in both patients' gallbladder tissue sections. To the best of our knowledge, C. bellii-associated cholecystitis has not been previously reported in a patient with prolonged corticosteroid therapy.

Effects of Anethum graveolens L. (dill) on Oocyte and Fertility of Adult Female Rats.

BACKGROUND: Our previous studies revealed Anethum graveolens L. caused some changes in female reproductive system that induced infertility. Therefore, in this study, oocyte changes as one of probable reasons of infertility were investigated. METHODS: In this study, 59 adult female rats were divided into 3 groups of control, low dose (0.5 g/kg) and high dose (5 g/kg) of dill seed aqueous extract (LDE and HDE) treated groups that were gavaged with 1 ml of each dose for 10 days (2 estrous cycles). Vaginal smears were prepared daily. Oocytes of superovulated animals were extracted and their morphometrical changes were measured (n = 5). Oocyte cell membrane glycoconjugates were stained with UEA, PNA, and DBA-FITC lectins (n = 5). Ultrastructural studies of oocytes were performed using TEM (n = 5). The number, weight, and crown-rump length of newborns were examined in three groups after mating with untreated males (n = 5). Data were analyzed using SPSS software. RESULTS: Results demonstrated that the duration of the estrous cycle, the diestrus phase and progesterone concentration in the experimental groups increased significantly compared to the control group (p < 0.05). Granulosa cells of corpus luteum in HDE-treated group were larger and clearer. The intensity reactions of galactose/Nacetylgalactoseamine terminal sugar of oocyte decreased insignificantly in experimental groups compared to the control group p > 0.05. Duration of mating to pregnancy increased and the weight and crown-rump length of newborns decreased in experimental groups significantly (p < 0.05). CONCLUSION: Dill seed aqueous extract can induce infertility without any effect on oocyte structure.

Effect of follicular fluid and platelet-activating factor on lactate dehydrogenase C expression in human asthenozoospermic samples.

BACKGROUND: Application of follicular fluid (FF) and platelet-activating factor (PAF) in artificial insemination improves sperm motility. Lactate dehydrogenase C (LDH-C) is a key enzyme for sperm motility. In this study, the effects of FF and PAF on the sperm motility index and LDH-C expression were investigated. Moreover, LDH-C expression was compared between asthenozoospermic and normozoospermic samples. METHODS: The expression of LDH-C was examined by quantitative real-time polymerase chain reaction (q-RT PCR) and western blotting after it was treated with optimized concentrations of FF and PAF in twenty asthenozoospermic samples. Also, LDH-C expression was evaluated in five normozoospermic samples. RESULTS: Samples with 75% FF and 100 nM of PAF had an increase in their percentages of progressive and slowly motile sperms and a decrease in their percentages of non-progressive and non-motile sperms. Moreover, LDH-C mRNA transcripts were not changed following PAF and FF treatment, and LDH-C protein was detected in highly progressive motile specimens treated with FF in the asthenozoospermic samples. Furthermore, LDH-C expression was more detectable in the normal sperms. CONCLUSION: Our results indicated that PAF had more beneficial effects than FF on sperm motility in the asthenozoospermic samples (P=0.0001), although the LDH-C expressions of the sperms were not changed significantly in both groups. We found no association between LDH-C expression and sperm motility after FF and PAF actions. This finding, however, requires further investigation. The fact that LDH-C protein was detected in the normozoospermic, but not asthenozoospermic, samples could be cited as a reason for the infertility in these patients.

Effects of L-carnitine and Pentoxifylline on the Activity of Lactate Dehydrogenase C4 isozyme and Motility of Testicular Spermatozoa in Mice.

BACKGROUND: Extracted sperm from the testis have poor motility. Moreover, their motility changes during their journey through epidydimis. Meanwhile, they face high concentration of L-carnitin. In addition, lactate dehydrogenase C4 (LDH-C4) gene disorders has been shown to cause impaired sperm motility, leading to infertility in male mice. The aim of this study was to evaluate sperm motility and LDH-C4 enzyme activity upon L-carnitine (LC) and Pentoxifylline (PTX) administrations in mice. METHODS: We extracted testicular sperm of 48 mice and divided them into three equal parts. One part was incubated with Ham's F10 medium (control), the other parts were treated with Ham's F10 containing LC and PTX with a final concentration of 1.76 mM, for 30 min at room temperature. Sperm motility was assessed according to the World Health Organization (WHO) criteria. Sperm LDH-C4 enzyme activity was measured by spectrophotometery method. Statistical analyses were performed using ANOVA and Fisher's LSD test, and a p-value less than 0.05 was considered as a statistically significant difference. RESULTS: Sperm motility increased after 30 min of incubation in LC- and PTX-treated group (p<0.001). LC and PTX administrations showed a significant increase in the LDHC4 enzyme activity of sperm compared to that of the controls after 30 min (P=0.04 and 0.01, respectively). CONCLUSION: The effects of LC and PTX on motility of sperm can be explained by an increase in LDH-C4 enzyme activity that may influence male fertility status. We suggest that LC as a non-toxic antioxidant is more suitable for use in assisted reproductive technique protocols than PTX.

Effects of L-carnitine and pentoxifylline on carbohydrate distribution of mouse testicular sperm membrane.

BACKGROUND: The glycoconjugate content of sperms indicates their physiological and fertility properties. Lectin reactivity is indicative of intact, capacitated, and acrosome-reacted sperms. In the epididymis, sperms experience maturation, glycoconjugate modification, and simultaneously, higher L-carnitine (LC) concentrations. The aim of this project was to evaluate the effects of LC and Pentoxifylline (PF) on the integrity, capacitation, and acrosomal reaction of sperms by studying their lectin reactivity. METHODS: Mouse testicular sperm samples were divided into three parts. Each sample was added Ham's F10 (control) or media containing 1.76 mM LC or PF. At 30 and 90 minutes after incubation, sperm motility was assessed. Peanut agglutinin (PNA), wheat germ agglutinin (WGA), and Concanavalin A (Con A) were used to detect non-acrosome-reacted, non-capacitated, and acrosome-reacted sperms, respectively and the frequency was evaluated by flow cytometry. Statistical analysis was performed using the ANOVA. RESULTS: Sperm motility increased after 30 and 90 minutes of incubation in the LC- and PF-treated cultures (P=0.001). LC administration created a significant increase in the percentage of the non-acrosome-reacted sperms compared to the control sperms after 30 and 90 minutes (P=0.02 and P=0.03, respectively). The frequency of the non-capacitated sperms in the LC-treated group increased compared to the control sperms after 30 minutes significantly (P=0.01). CONCLUSION: Although the administration of LC and PF enhanced sperm motility, LC also impacted glycoconjugates on the sperm surface. Glycoconjugates are involved in the interaction between the sperm and the zona pellucida and subsequently fertilization, thereby probably influencing the male fertility state.

Effects of L-carnitine and L-acetyl-carnitine on testicular sperm motility and chromatin quality.

BACKGROUND: Sperm cells extracted from testes (TESE) have poor chromatin quality and motility. Various substances are used in the laboratory to increase sperm motility and improve the ART outcomes; however, there are few research which considered improving both sperm motility and chromatin quality. OBJECTIVE: The aim of this investigation was to evaluate the improvement of the testicular sperm motility and chromatin quality exposed to L-carnitine (LC) and L-acetyl-carnitine (LAC), which are normally concentrated in testis and epididymis, compared with Pentoxifylline (PF), which used for sperm motility enhancement in IVF procedures. MATERIALS AND METHODS: TESE samples from 30 male mice divided into four parts. The sperm samples were added to Ham' F10 (control) or the media contained 1.76mM of LC, LAC or PF), then, the samples were kept in the room temperature for 30, 90 and 180 min. At each time step, sperm motility and chromatin quality were assessed. Chromatin quality was evaluated by chromomycin A3 and aniline blue. Statistical analysis was performed using one way analysis of variance (ANOVA). A p-value less than 0.05 were accepted as a statistically significant difference. RESULTS: The results showed LC, LAC and PF significantly increased the sperm motility. However, sperm chromatin quality only improved significantly by administration of LC and LAC. CONCLUSION: Administration of LC and LAC to the testicular sperm samples can lead to improve both sperm motility and chromatin quality. It may be because they can mimic in vivo sperm condition during late spermatogenesis.

Effects of curcumin on the dorsal root ganglion structure and functional recovery after sciatic nerve crush in rat.

BACKGROUND: Curcumin has protective effects on a variety of CNS injury models. Our purpose was to assess the effects of curcumin on the dorsal root ganglion (DRG) and functional recovery in sciatic nerve crush (SNC) of rats. METHODS: The fifth lumbar DRG was removed 28 days after SNC. Sciatic functional index (SFI) and also the DRG structure were evaluated, using stereological methods to assess the effects of curcumin. RESULTS: The volume and number of A- and B-cells were reduced in the SNC as compared with the sham-operated animals. In curcumin treated rats, the volume decreased but in a lesser extent and it was ∼10% and ∼15% higher on the average in comparison with the non-treated SNC (P<0.04). In curcumin treated rats, the number of cells also decreased but in a lesser extent and it was ∼17% and ∼36% higher in comparison with the non-treated crushed animals (P<0.02). The surface area of A- and B-cells reduced in the SNC and SNC plus curcumin in comparison with the sham-operated rats (P<0.01). The number of satellite cells was reduced in the SNC as compared with the sham-operated animals. In curcumin treated rats, the number of satellite cells decreased but in a lesser extent and the number of satellite cells was ∼19% higher in comparison with the non-treated (P<0.02). SFI test shows that the motor function of the rats treated with curcumin significantly improved at 21st days after sciatic nerve crush. CONCLUSION: The results demonstrate that curcumin promotes the functional recovery and has a neuroprotective effect on the DRG after sciatic nerve injury in the rats.

Ameliorative effects of curcumin on the structural parameters of seminiferous tubules and Leydig cells in metronidazole-treated mice: a stereological approach.

Metronidazole (MTZ), an anti-parasitic drug, induces negative effects on the testis. Curcumin exhibits antioxidant properties and anti-tumor properties. The aim was to evaluate negative effects of seminiferous tubules and Leydig cells by MTZ and ameliorative effects of curcumin. Balb/c mice were divided into six groups. The control, second, third, fourth and fifth, and sixth groups were administrated distilled water, high doses of MTZ (500 mg/kg/day), MTZ (500 mg/kg/day) +100 mg/kg/day curcumin, therapeutic doses of MTZ (165 mg/kg/day), MTZ (165 mg/kg/day) +100 mg/kg/day curcumin, and 100 mg/kg/day curcumin, respectively. The data revealed significant reduction in tubule volume, length and diameter and germinal epithelium height, and increase the number of Leydig cells in MTZ-treated (high or therapeutic doses) animals. Combined treatment of curcumin with high or therapeutic doses of MTZ ameliorated the increase in number of Leydig cell. Ameliorative effects of curcumin on the other above-mentioned parameters were observed in mice receiving therapeutic doses of MTZ. Leydig cell or nucleus volume and interstitial tissue volume did not show any significant difference in the MTZ-treated mice. It can be concluded that metronidazole can changes structural parameters of the tubules and number of Leydig cells and curcumin can ameliorate these effects.

Ameliorative effects of curcumin on the seminiferous epithelium in metronidazole-treated mice: a stereological study.

Metronidazole (MTZ) has negative effects on sperm analysis and testis structure. Curcumin is the principal curcuminoid found in turmeric and exhibits antitumor, anti-inflammatory, and anti-infectious activities with low toxicity. To evaluate stereological changes of seminiferous germinal epithelium by MTZ and ameliorative effects of curcumin, Balb-c mice were divided into 6 groups. The control, second, third, fourth, fifth, and sixth groups were administrated distilled water, high doses of MTZ (500 mg/kg/day), MTZ (500 mg/kg/day) + 100 mg/kg/d curcumin, therapeutic doses of MTZ (165 mg/kg/day), MTZ (165 mg/kg/day) + 100 mg/kg/day curcumin, and 100 mg/kg/day curcumin, respectively. Testis weight, testis volume, total epithelial volume, spermatocytes, and spermatid number showed a significant reduction in MTZ-treated (high or therapeutic doses) animals in comparison with the controls. Curcumin can protect spermatocytes after high or therapeutic doses of MTZ treatment. But curcumin is able to protect the other parameters only in the mice receiving therapeutic doses of MTZ. The total number of sertoli cells and spermatogonia did not show any significant difference in the mice that received MTZ. MTZ can reduce germinal epithelium volume and the number of spermatocytes and spermatids. Ameliorative effects of curcumin can mainly be observed in the mice receiving therapeutic doses of MTZ.