Impact of 5'-AMP-activated protein kinase (AMPK) on Epithelial Sodium Channels (ENaCs) in human sperm.

Hyperpolarization is associated with decreased intracellular Na+ concentration through the closure of the epithelial Na+ channels (ENaCs) during capacitation. 5'-AMP-activated protein kinase (AMPK) is involved in the regulation of Na+ transport by reducing ENaC-ß abundance in the plasma membrane in somatic cells. However, it is not known whether AMPK acts on ENaCs in sperm. The aim of the present study was to analyze the role of AMPK activation in the regulation of ENaC and to examine its relationship with capacitation-associated hyperpolarization of human sperm. Human sperm were treated with AICAR (AMPK activator) in non-capacitating and capacitating conditions. AMPK activity and ENaC-ß concentration were evaluated by ELISA. Flow cytometry was used to measure tyrosine phosphorylation, hyperpolarization, intracellular Na+ concentration and acrosome reaction. Immunofluorescence staining was carried out to analyze the distribution of ENaC-ß and CD46 in sperm. We found that induction of capacitation triggered AMPK phosphorylation. AMPK activation by AICAR increased tyrosine phosphorylation. AICAR decreased ENaC-ß levels, mainly localized at the principal-piece of the flagellum, resulting in lower intracellular Na+ concentration and increased hyperpolarization of the plasma membrane. Altogether, these data provide evidence that AMPK activation is involved in capacitation-associated hyperpolarization by reducing ENaC abundance in human sperm.

Examination of clinical data and semen analysis results of patients undergoing orchiectomy for testicular tumor.

OBJECTIVE: Testicular tumor constitutes 1% of male neoplasms. Infertility can be determined in patients with testicular tumors before orchiectomy due to the deterioration of spermatogenesis. The aim of this study was to show the clinical, radiological, and pathological characteristics and spermiogram results of patients with testicular tumor and their relationship with each other. METHODS: The data of patients who underwent orchiectomy due to testicular tumor between 2016 and 2019 were reviewed retrospectively. These data included sociodemographic data of the patients, pretreatment spermiogram characteristics, level of serum tumor markers, characteristics of the ultrasonography, type of orchiectomy, and histopathological examination. RESULTS: This study included 53 male patients, with a mean age of 33.51±12.86 years. The mean levels of all tumor markers were above the reference levels. The mean tumor size was 34.68±23.32 mm. Multiple localizations and microlithiasis were detected in 11.3 and 13.2% of the tumors, respectively. The most common masses were hypoechoic (n=37; 69.8%) and hypervascular (n=47; 81%). Spermiogram and cryopreservation were performed in 29 (54.7%) of 53 patients preoperatively. The mean sperm concentration before orchiectomy was 24.21×106 /mL and group A sperm motility 0.79%, group B sperm motility 39.10%, group C sperm motility 9.83%, and group D sperm motility 22.69% in testicular tumors. CONCLUSION: Spermatogenesis adversely affected before the treatment due to local and systemic effects of testicular cancer. Fertility expectations can be increased in the subsequent years by semen analysis and referral to cryopreservation.

Examination of clinical data and semen analysis results of patients undergoing orchiectomy for testicular tumor

SUMMARY OBJECTIVE: Testicular tumor constitutes 1% of male neoplasms. Infertility can be determined in patients with testicular tumors before orchiectomy due to the deterioration of spermatogenesis. The aim of this study was to show the clinical, radiological, and pathological characteristics and spermiogram results of patients with testicular tumor and their relationship with each other. METHODS: The data of patients who underwent orchiectomy due to testicular tumor between 2016 and 2019 were reviewed retrospectively. These data included sociodemographic data of the patients, pretreatment spermiogram characteristics, level of serum tumor markers, characteristics of the ultrasonography, type of orchiectomy, and histopathological examination. RESULTS: This study included 53 male patients, with a mean age of 33.51±12.86 years. The mean levels of all tumor markers were above the reference levels. The mean tumor size was 34.68±23.32 mm. Multiple localizations and microlithiasis were detected in 11.3 and 13.2% of the tumors, respectively. The most common masses were hypoechoic (n=37; 69.8%) and hypervascular (n=47; 81%). Spermiogram and cryopreservation were performed in 29 (54.7%) of 53 patients preoperatively. The mean sperm concentration before orchiectomy was 24.21×106 /mL and group A sperm motility 0.79%, group B sperm motility 39.10%, group C sperm motility 9.83%, and group D sperm motility 22.69% in testicular tumors. CONCLUSION: Spermatogenesis adversely affected before the treatment due to local and systemic effects of testicular cancer. Fertility expectations can be increased in the subsequent years by semen analysis and referral to cryopreservation.

Effect simultaneous delivery with P-glycoprotein inhibitor and nanoparticle administration of doxorubicin on cellular uptake and in vitro anticancer activity.

Multidrug resistance (MDR) is the most common problem of inadequate therapeutic response in tumor cells. Many trials has been developed to overcome drug efflux by P-glycoprotein (P-gp). For instance, co-administration of a number of drugs called chemosensitizers or MDR modulators with a chemotherapeutic agent to inhibit drug efflux. But for optimal synergy, the drug and inhibitor combination may need to be temporally colocalized in the tumor cells. In this study, we encapsulated the Ver and Dox in PLGA nanoparticles to inhibit the P-gp drug efflux in breast cancer. Moreover, the effect of either Dox solution (DoxS), Dox nanoparticles (DoxNP), DoxS + VerS, DoxNP + VerS, DoxNP + VerNP or Dox-VerNP was evaluated. It was found that co administration of DoxNP with VerNP (70.76%) showed similar cellular uptake of Dox to Dox/Ver combination solution (70.62%). However it is observed that DoxNP + VerNP has the highest apoptotic activity (early apoptotic 13.52 ± 0.06%, late apoptotic 53.94 ± 0.15%) on human breast adenocarcinoma (MCF 7) cells. Hence, it is suggested that DoxNP + VerNP is a promising administration for tumor therapy.

Investigation of transfer results of human embryos that were vitrified and thawed at the cleavage, morula and blastocyst stages.

The objective of this study was to compare the rates of clinical pregnancy after the transfer of vitrified and thawed human embryos on days 3, 4 and 5 of embryonic development. In this retrospective study, the results of 148 embryo transfer cycles, using embryos frozen and thawed over the 3-year period between January 2016 and December 2018 at the Gülhane Training and Research Hospital Department of Gynecology and Obsterics Reproductive Medical Center of the University of Health Sciences, Ankara, Turkey were examined. Following embryo transfer - including 29 dissolved embryos frozen on day 3, 80 frozen on day 4, and 39 frozen on day 5 - results were examined in terms of clinical pregnancy rates. In this study, across all three groups, no significant differences were observed in terms of patient age, the number of oocytes collected, infertility reasons, the number of embryos dissolved, transfer day, or the number of embryos transferred. According to the transfer day, the rates of clinical pregnancy and ongoing pregnancy were significantly higher for embryos frozen on day 4 and transferred on day 5. Significantly higher rates of pregnancy and live birth were determined during in vitro fertilization (IVF) treatment with the freezing of human embryos on day 4 and the transfer of those embryos on day 5.

Effects of triethylene glycol dimethacrylate (TEGDMA) on the odontoclastic differentiation ability of human dental pulp cells.

OBJECTIVES: The primary purpose of this study was to examine the effects of triethylene glycol dimethacrylate (TEGDMA) on odontoclastic differentiation in the dental pulp tissue. MATERIAL AND METHODS: The effects of different TEGDMA dosages on the odontoclastic differentiation capability of dental pulp cells were analyzed in vitro using the following methodologies: i) flow cytometry and tartrate-resistant acid phosphatase (TRAP) staining; ii) apoptotic effects using Annexin V staining; iii) mRNA expression of osteoprotegerin (OPG) and receptor activator of nuclear factor (NF)-kB ligand (RANKL) genes by quantitative Real-time PCR (qRT-PCR); and iv) OPG and RANKL protein expression by enzyme-linked immunosorbent assay (ELISA). RESULTS: TEGDMA caused relatively less odontoclastic differentiation in comparison with the control group; however, odontoclastic differentiation augmented with increasing doses of TEGDMA (p<0.05). The mRNA and protein expression of OPG was lower in TEGDMA treated pulp cells than in the control group (p<0.05). While the mRNA expression of RANKL remained unchanged compared to the control group (p>0.05), its protein expression was higher than the control group (p<0.05). In addition, TEGDMA increased the apoptosis of dental pulp cells dose dependently. CONCLUSIONS: TEGDMA reduced the odontoclastic differentiation ability of human dental pulp cells. However, odontoclastic differentiation ratios increased proportionally with the increasing dose of TEGDMA.

Histological Characteristics of Ligaments between Middle Ear and Temporomandibular Joint.

OBJECTIVES: To examine histological aspects of the ligaments between the middle ear and temporomandibular joint and suppose a theoretical role of their structural characteristics on mobility of mallear ossicle. METHODS: The ligaments were obtained by microdissection of middle cranial fossa on both sites of 15 cadavers fixed in formalin solution and were sectioned longitudinally (7-10 mum thickness). The sections were stained with Verhoff's Van Gieson's stain (VVG) for demonstration of elastic fibers and visualized at X2.5 and X10 magnifications under light microscopy. RESULTS: Anterior mallear ligament (AML) and sphenomandibular ligaments (SML) were consisted of collagen fibres in analyzed specimens. The discomallear ligament (DML) was constituted of rich collagenous fibres. One specimen of DML harvested between petrotympanic fissure and retrodiscal-capsular intersection site contained elastic fibers dispersed in cotton-bowl appearance between collagen fibers. In the light of functional tests performed in previous studies, it could be extrapolated that presence of elastic fibers in the DML may prevent excessive forces conducted to mallear head by elongation of elastic fibers. CONCLUSIONS: Collagenous fibres have no ability to stretch along their axis which may lack compensatory mechanism to prevent mallear head mobility.

Perindopril, atenolol, and amlodipine prevent aortic ultrastructural changes in rats exposed to ethanol.

BACKGROUND: The effects of perindopril, an angiotensin-converting enzyme inhibitor, atenolol, a beta adrenergic receptor blocker, and amlodipine, a calcium channel blocker, were investigated in chronic alcohol administered rats. MATERIAL/METHODS: Adult male Wistar rats (240-320 g) were used in the present study. Alcohol was given to rats on a modified liquid diet for 21 days. Perindopril (2.5 and 5 mg/kg), atenolol (5 and 10 mg/kg), and amlodipine (5 and 10 mg/kg) were injected into rats in different groups intraperitoneally for 21 days. Control rats were pair fed an isocaloric liquid diet containing sucrose as a caloric substitute for alcohol. Saline was injected into the control rats for 21 days. The hearts were removed after the rats were anesthetized by ether, and 1-mm3 samples from the ascending aortas were fixed. Five fields per aorta were examined and photographed with a transmission electron microscope. Blood alcohol levels were also measured spectrophotometrically. RESULTS: Daily alcohol consumption of the rats was in the range of 12.09-15.50 g/kg. Blood alcohol concentrations were 145.63 mg/dl on the 21st day of alcohol consumption. Chronic alcohol consumption caused some marked aortic wall injuries. Perindopril, atenolol, and amlodipine at high doses, but not low doses, produced some significant beneficial effects on alcohol-induced aortic wall damage. CONCLUSIONS: These results imply that perindopril, atenolol, and amlodipine may have protective effects on heavy chronic alcohol consumption-induced aortic wall injury in rats only in high doses.

Opinions of infertile Turkish women on gamete donation and gestational surrogacy.

OBJECTIVE: To determine the approval levels of infertile Turkish women concerning gamete donation and gestational surrogacy. DESIGN: Opinion survey. SETTING: Assisted reproductive treatment center at Gulhane Military Medical Academy. PATIENT(S): 368 women who had applied for infertility treatment. INTERVENTION(S): The patients were asked to answer a questionnaire that included questions about the patient's sociodemographic status, previous medical history with infertility treatment, and opinions on gamete donation and gestational surrogacy. MAIN OUTCOME MEASURE(S): Opinions of patients concerning gamete donation and gestational surrogacy. RESULT(S): Our data in this first study on infertile Turkish women show that some patients approve of gamete donation (23.3% for accepting oocytes and 3.4% for accepting sperm) and gestational surrogacy (15.1%). CONCLUSION(S): Donation and surrogacy are alternate treatments for the serious condition called infertility. As there are patients who would like to use these treatments, each of these patients should have the right to try any of them.

The prevention of myocardial ultrastructural changes by perindopril, atenolol and amlodipine in chronic alcohol administered rats.

The effects of perindopril, an angiotensin converting enzyme inhibitor, atenolol, a beta adrenergic receptor blocker and amlodipine, a calcium channel blocker were investigated in chronic alcohol administered rats. Adult male Wistar rats (240-320 g) were used in the present study. Alcohol was given to rats by a modified liquid diet for 21 days. Perindopril (2.5 and 5 mgkg(-1)), atenolol (5 and 10 mg kg(-1)) and amlodipine (5 and 10 mg kg(-1)) were injected to rats in different groups intraperitoneally for 21 days. Control rats were pair fed by an isocaloric liquid diet containing sucrose as a caloric substitute for alcohol. Saline was injected to control rats for 21 days. Rats were anesthetized with ether. Their hearts were removed and 1 mm3 samples from left ventricles were fixed. Five fields per heart were examined and photographed with transmission electron microscope. Blood alcohol levels were also measured spectrophotometrically. Daily alcohol consumption of the rats was in a range of 12.09-15.5 g kg(-1). Blood alcohol concentrations were found as 145.63 mg dl(-1) at 21st day of alcohol consumption. Chronic alcohol consumption caused some marked myocardial injuries. Perindopril and atenolol but not amlodipine produced some significant beneficial effects on alcohol-induced myocardial damages. Our results imply that perindopril and atenolol but not amlodipine have protective effects on heavy chronic alcohol consumption-induced myocardial injury in rats.