Phoenix dactylifera L. pollen versus pentoxifylline on improvement of sperm parameters in idiopathic male infertility: A randomized clinical trial.

ETHNOPHARMACOLOGICAL RELEVANCE: Phoenix dactylifera L. pollen is the male reproductive dust of palm flowers known as a natural product that is considered a strong stimulant of sexual potency and fertility in Iranian traditional medicine (ITM). In this regard, no evidence-based medications are empirically prescribed to treat IMI. However, applying traditional medicine for the treatment of male infertility has attracted more attention in recent years. AIM OF THE STUDY: Phoenix dactylifera L. pollen was compared with pentoxifylline (PTX) to evaluate its efficacy on sperm parameters. MATERIALS AND METHODS: During this parallel randomized controlled trial, 80 adult men with asthenozoospermia, oligozoospermia, or teratozoospermia (age 20-35 years) were enrolled. In two separate groups of participants with a 1:1 ratio, participants received either 6 g of Phoenix dactylifera L. pollen powder daily or 400 mg of PTX tablets daily for 90 days. We measured the sperm parameters as well as the serum sex hormones in the sample. ANCOVA and t-tests were used to compare groups. RESULTS: There was no significant difference between the study groups in terms of baseline characteristics or demographic characteristics. According to the results, participants who took Phoenix dactylifera L. pollen powder had significantly improved sperm concentration (p = 0.016), morphology (p = 0.029), sperm counts (p = 0.012), progressive motility (p = 0.016), total motility (p = 0.018), and reduced immotile sperms (p = 0.014) compared to those who took PTX. CONCLUSIONS: In light of these results, Phoenix dactylifera L. pollen is recommended as a treatment factor for ameliorating IMI by enhancing sperm functional capacity and semen parameters.

Predicting the survival of kidney transplantation: design and evaluation of a smartphone-based application.

BACKGROUND: Prediction of graft survival for Kidney Transplantation (KT) is considered a risky task due to the scarcity of donating organs and the use of health care resources. The present study aimed to design and evaluate a smartphone-based application to predict the survival of KT in patients with End-Stage Renal Disease (ESRD). METHOD: Based on the initial review, a researcher-made questionnaire was developed to assess the information needs of the application through urologists and nephrologists. By using information obtained from the questionnaire, a checklist was prepared, and the information of 513 patients with kidney failure was collected from their records at Sina Urological Research Center. Then, three data mining algorithms were applied to them. The smartphone-based application for the prediction of kidney transplant survival was designed, and a standard usability assessment questionnaire was used to evaluate the designed application. RESULTS: Three information elements related to the required data in different sections of demographic information, sixteen information elements related to patient clinical information, and four critical capabilities were determined for the design of the smartphone-based application. C5.0 algorithm with the highest accuracy (87.21%) was modeled as the application inference engine. The application was developed based on the PhoneGap framework. According to the participants' scores (urologists and nephrologists) regarding the usability evaluation of the application, it can be concluded that both groups participating in the study could use the program, and they rated the application at a "good" level. CONCLUSION: Since the overall performance or usability of the smartphone-based app was evaluated at a reasonable level, it can be used with certainty to predict kidney transplant survival.

Overexpression of long intergenic noncoding RNAs in bladder cancer: A new insight to cancer diagnosis.

There is increasing evidence that show long noncoding RNAs including long intergenic noncoding RNAs (lincRNA) play a pivotal regulatory role in the biological processes. Differential expression of lincRNAs can be utilized for cancer diagnosis, prognosis, and targeted therapy. Little is known about their expressions in urothelial tumors. Concerning the potential role of lincRNAs in cancer development, we aimed to investigate the expression levels of LINC00958 and DNM3OS in bladder cancer. Fifty tumor and 50 adjacent non-tumor tissue samples along with their clinicopathological parameters were obtained from bladder cancer patients. Expressions of LINC00958 and DNM3OS were analyzed by Real-time PCR. ROC curve analysis was used to evaluate the diagnostic power of LINC0095 and DNM3OS for BC. Expression level of LINC00958 was considerably increased in cancerous tissues (P < 0.001) and in correlation with cigarette smoking (P = 0.043). DNM3OS expression was higher in the tumor tissues than normal tissues (P < 0.001) and showed a significant association with age (P = 0.038). By using the ROC curve, the diagnostic power of LINC00958 and DNM3OS transcript levels in bladder cancer were estimated to be 87% and 75%, respectively. Our findings offer some important intuitions into the oncogenic role of LINC00958 and DNM3OS in bladder cancer and suggest that they can be candidate biomarkers and may provide new approaches for the diagnosis and therapy if being validated in a larger sample size of clinical samples as well as functional studies.

The influence of socioeconomic factors on deceased organ donation in Iran.

Background: There is a large gap between the number of patients on organ waiting lists and the number of available organs for donation. This study investigated the socioeconomic factors in Iran that influenced decisions for organ donation among the families of brain-dead donors. Methods: This retrospective cross-sectional study was performed among the families of 333 organ donors in Iran. Two trained researchers interviewed family members about the donor's age, sex, cause of brain death, education level, marital status, number of children, history of addiction, the financial status of the donor's family, and reasons for which they considered refusing organ donation. Results: The mean age of the donors was 37.23±16.59 years. During 2017-2019, significant differences were found according to income (P<0.001), marital status (P<0.001), sex (P=0.04), and occupation (P=0.04). More than half of the organ donors were of low socioeconomic status, and nearly half were the sole income earners of large families. Trauma was the most common cause of death (44.6%). The most common reasons for which the families considered refusing organ donation were unfamiliarity with the concept of brain death, denial, and the expectation of a miracle. Conclusions: The donor's socioeconomic status and availability of social services, such as insurance coverage, psychological services, and mourning therapy courses, play an important role in organ donation. Adequate support for the deceased's family after organ donation is imperative.

Antimicrobial susceptibility testing and molecular characterization of Neisseria gonorrhoeae in Tehran, Iran.

Gonorrhea is a sexually transmitted infection occurring worldwide. Antimicrobial resistance (AMR) surveillance in Neisseria gonorrhoeae and associated molecular epidemiological studies are crucial to ascertain the spread of antibiotic-resistant and developing the local treatment guidelines. This study was performed to determine the antimicrobial susceptibility testing (AST) and molecular epidemiology of N. gonorrhoeae isolates in Tehran, Iran. During 1 July 2018-30 July 2020, a total of 500 urogenital (468 endocervical, 32 urethral) swabs were collected from patients with signs and symptoms of genitourinary infections presenting to two women's hospitals and one health center located center and south of Tehran. Specimens were cultured and examined for the presence of N. gonorrhoeae isolates by biochemical tests. MIC Test Strip determined the MICs of ceftriaxone, azithromycin, and ciprofloxacin. Neisseria gonorrhoeae multiantigen sequence typing (NG-MAST) was also performed. A total of 38 N. gonorrhoeae isolates were identified. The proportions of resistant N. gonorrhoeae isolates were as follows: ceftriaxone (MIC ≥0.125 µg/mL) 10.5% (4/38), azithromycin (MIC >1 µg/mL) 34% (13/38), and ciprofloxacin (MIC ≥1 µg/mL) 31.5% (12/38). In total, 25 different NG-MAST STs were identified. The STs comprised 1-4 isolates each, and the predominant ST was ST266 (n = 4). Our study demonstrates a diverse gonococcal population with high rates of resistance to azithromycin and evidence of resistance to ceftriaxone. The results have potential implications for antibiotic choice for the gonococcal treatment and highlight the need to broaden gonococcal AMR monitoring in Iran.

Evaluation of Thymic Output and Regulatory T Cells in Kidney Transplant Recipients with Chronic Antibody-Mediated Rejection.

BACKGROUND: Regulatory T cells (Tregs) and recent thymic emigrants (RTEs) have an essential role in the regulation of allogeneic immune responses. However, their mechanisms of action in chronic antibody-mediated rejection (cAMR) are still unclear. In this study, we aimed to compare Treg and RTE levels between stable graft function (SGF) patients and cAMR subjects after kidney transplantation. METHOD: Mononuclear cells (MNs) were separated from peripheral blood, and flow cytometry analysis was performed for detection of CD4+ and CD25high as Treg markers and CD4+, CD31+, and CD45RA+ as RTE immunophenotyping markers. RESULT: The level of peripheral Treg cells was significantly lower in cAMR subjects in comparison to stable graft function patients. Moreover, SGF patients who had received cyclosporine A had a higher level of Treg in comparison to the tacrolimus recipients. Nevertheless, the RTE level between SGF and cAMR patients did not show any significant differences. CONCLUSION: It seems that Treg cells are significantly associated with transplant outcomes in cAMR patients, and prescribed immunosuppressive drugs can influence the frequency of this crucial subset of T cells. Although these drugs are beneficial and inevitable for allograft maintenance, more investigations are needed to elucidate their complete effects on different immune cell subsets which some of them like Tregs are in favor of transplant tolerance. Besides, the thymic output is seemingly not a beneficial biomarker for predicting cAMR; however, more in vivo and in vitro studies are needed for revealing the precise role of Tregs and RTEs in the transplantation context.

Living versus deceased kidney transplantation: Comparison of complications.

INTRODUCTION: It is assumed that the outcome of kidney transplantation from living donors is more favorable than deceased donors. However, over the years there has been an overall improvement in transplant survival from both living and deceased donors. In this article we have evaluated and compared the most recent results in living and deceased donor kidney transplantations. PATIENTS AND METHODS: Four hundred and forty six patients underwent kidney transplantation in our center from September 2009 to March 2014. The patients divided in two groups living (group A) and deceased (group B) donor transplantation groups. The patients were followed until September 2016. Acute rejection, graft survival, delayed graft function, renal artery thrombosis, graft nephrectomy, ureterocutaneous fistula, postoperative hypertension, mortality, hospital stay, hyperlipidemia, post transplantation diabetes and lymphocele rate measured and compared in two groups. RESULTS: Most variables were not different between the two groups except lymphocele and delayed graft function. Lymphocele was more prevalent in group A (13.8% vs 3.1%, p-value = 0.02) and delayed graft function results were more desirable in living donor transplantation group (group B). CONCLUSION: Although, delayed graft function was less common in transplantation from living donors, short, and long term graft survival was not significantly different in this study.

Isolation, identification and differentiation of human spermatogonial cells on three-dimensional decellularized sheep testis.

Improvement of in vitro culture methods of Spermatogonial Stem Cells (SSCs) is known to be an effective procedure for further study of the process of spermatogenesis and can offer effective therapeutic modality for male infertility. Tissue decellularization by providing natural 3D and extracellular matrix (ECM) conditions for cell growth can be an alternative procedure to enhance in vitro culture conditions. In the present study, the testicular tissues were taken from brain death donors. After enzymatic digestion, the tissue cells were isolated and cultured for four weeks. Then the identity of the SSCs was confirmed using anti-GFRα1 and anti-PLZF antibodies via immunocytochemistry (ICC). The differentiation capacity of SSCs were evaluated by culture of them on a layer of decellularized testicular matrix (DTM) prepared from sheep testis, as well as under two-dimensional (2D) culture with differentiation medium. After four and six weeks of the initiation of differentiation culture, the pre-meiotic, meiotic and post- meiotic genes at the mRNA and protein levels was examined via qPCR and ICC methods, respectively. The results showed that pre-meiotic, meiotic and post-meiotic genes expressions were significantly higher in the cells cultured in DTM substrate (P ≤ 0.01).The present study indicated that, the natural structure of ECM prepare the suitable conditions for further study of the spermatogenesis process in the in vitro and contributes to the maintenance and treatment of male infertility.

An application of the theory of planned behavior to self-care in patients with hypertension.

BACKGROUND: Self-care behaviors and positive changes in lifestyle are essential for successful hypertension control. We used a behavioral model based on the theory of planned behavior to assess which factors influence self-care behaviors for controlling hypertension. METHODS: In this cross-sectional study, five hundred patients with at leastaone-year history of diagnosed hypertension participated in this study. The data collection tool was designed based on the theory of planned behavior. Structural equation modeling was used to estimate the main parameters. RESULTS: For self-care behaviors, ninety-six (19.2%) and forty-five (9.1%) participants had good knowledge and acceptable behavior(≥8 out of 10 points). Having perceived behavioral control regarding quitting smoking and alcohol intake was associated with the patient's intention and behavior [b:1.283 ± .095 and b:1.59 ± .014 (p < .001)]. Having perceived behavioral control over the other self-care behaviors had a positive effect on the intention in female patients [b: .885 ± .442 (p = .045)]. Subjective norms had a positive effect on behavioral intention in younger patients [b:4.52 ± 2.24 (P = .04)]. CONCLUSIONS: Group-specific behavioral barriers are important when improving self-care behaviors in patients with hypertension. Perceived control over self-care behaviors is more important in vulnerable patients, such as the elderly and women.

Living or deceased-donor kidney transplant: the role of psycho-socioeconomic factors and outcomes associated with each type of transplant.

BACKGROUND: Kidney transplant improves patients' survival and quality of life. Worldwide, concern about the equality of access to the renal transplant wait-list is increasing. In Iran, patients have the choice to be placed on either the living or deceased-donor transplant wait-list. METHODS: This was a prospective study performed on 416 kidney transplant recipients (n = 217 (52.2%) from living donors and n = 199 (47.8%) from deceased donors). Subjects were recruited from four referral kidney transplant centers across Tehran, Iran, during 2016-2017. The primary outcome was to identify the psycho-socioeconomic factors influencing the selection of type of donor (living versus deceased). Secondary objective was to compare the outcomes associated with each type of transplant. The impact of psycho-socioeconomic variables on selecting type of donor was evaluated by using multiple logistic regression and the effect of surgical and non-surgical variables on the early post-transplant creatinine trend was assessed by univariate repeated measure ANOVA. RESULTS: Based on standardized coefficients, the main predictors for selecting living donor were academic educational level (adjusted OR = 3.25, 95% CI: 1.176-9.005, p = 0.023), psychological status based on general health questionnaire (GHQ) (adjusted OR = 2.46, 95% CI: 1.105-5.489, p = 0.028), and lower monthly income (adjusted OR = 2.20, 95% CI: 1.242-3.916, p = 0.007). The waiting time was substantially shorter in patients who received kidneys from living donors (p < 0.001). The early post-transplant creatinine trend was more desirable in recipients of living donors (ß = 0.80, 95% CI: 0.16-1.44, p-value = 0.014), patients with an ICU stay of fewer than five days (ß = - 0.583, 95% CI: - 0.643- -0.522, p-value = < 0.001), and those with less dialysis duration time (ß = 0.016, 95% CI: 0.004-0.028, p-value = 0.012). Post-operative surgical outcomes were not different across the two groups of recipients (p = 0.08), however, medical complications occurred considerably less in the living-donor group (p = 0.04). CONCLUSION: Kidney transplant from living donors was associated with shorter transplant wait-list period and better early outcome, however, inequality of access to living donors was observed. Patients with higher socioeconomic status and higher level of education and those suffering from anxiety and sleep disorders were significantly more likely to select living donors.

Tumor-derived urinary exosomal long non-coding RNAs as diagnostic biomarkers for bladder cancer.

Bladder cancer (BC) is the sixth most common malignancy in men and 17th in women. Exosomal long non-coding RNAs (lncRNAs) have been defined as a novel biomarker for BC. The aim of this study is to evaluate the clinical significance of urine exosomal PVT-1, ANRIL and PCAT-1 as a biomarker in BC patients with tumors classified as T1 or T2. Exosomes were isolated from urine of BC patients and healthy donors, then characterized according to their shape, size, and exosome markers by Electron Microscopy, Dynamic light scattering, and Western blotting. Exosomal lncRNAs extraction was done to determine the expression levels of PVT-1, ANRIL and PCAT-1 by qRT-PCR. ANRIL and PCAT-1 expression was significantly higher in BC patients compared to normal subjects. To evaluate the performance of the identified lncRNAs for BC detection, we performed ROC curves analysis. The diagnostic accuracy of ANRIL and PCAT-1, measured by AUC, was 0.7229 (sensitivity = 46.67 % and specificity = 87.5 %) and 0.7292 (sensitivity = 43.33 % and specificity = 87.5 %). Transcript levels of lncRNAs in urinary exosomes are potential diagnostic biomarkers in bladder cancer.

Early Conversion to Tacrolimus Vs Cyclosporine Continuation in Normally Functioning Kidney Allograft: A Single-Center Study.

This study evaluated the effectiveness of early pre-emptive conversion from cyclosporine to tacrolimus in kidney transplant patients with normal graft function and in the absence of adverse effects of the initial cyclosporine. A historical cohort study of 166 patients who received deceased-donor kidney transplant between 2011 to 2017 was conducted. All the patients had been treated with cyclosporine (Sandimmune®) during their immediate post-transplantation period. At the time of hospital discharge, the patients were divided into 2 groups: patients with continued cyclosporine (Sandimmune®) treatment (n = 125) and the patients whose treatments converted from cyclosporine to tacrolimus (Prograf®) at discharge (n = 41). The 1-year graft function (p = 0.074), acute rejection (p = 0.566), and graft loss (p = 0.566) were not significantly different between two groups. The patients on tacrolimus had lower levels of cholesterol (p = 0.002) and diastolic blood pressure (p = 0.015). The long-term follow-up showed no significant difference in graft loss (p = 0.566). The patients received tacrolimus had higher all-cause mortality within the first year posttransplantation (p = 0.002) as well as long-term follow-up (p = 0.001). The continuation of initial cyclosporine might be a good option when the graft function is acceptable and the adverse effects are absent.

Therapeutic effects of supplementation with Curcuminoids in critically ill patients receiving enteral nutrition: a randomized controlled trial protocol.

BACKGROUND: Curcuminoids are dietary polyphenols that can improve health indices through different mechanisms such as anti-inflammatory, antioxidant and immunoregulatory properties. Due to the lack of evidences on the efficacy of curcuminoids in critically ill patients, this study was designed to investigate the effects of short-term curcuminoids supplementation on inflammatory, oxidative stress and adipokine indices as well as nutritional and clinical status in Traumatic Brain Injury (TBI) patients admitted in the Intensive Care Unit (ICU). METHODS: The present trial will be performed in the ICU of Sina and Shohadaye Tajrish hospitals of Tehran, Iran. Sixty-two critically ill patients with TBI will be enrolled based on the eligibility criteria. The patients will be randomly assigned into two groups. For 7 days, they will received either 500 mg curcuminoids in combination with 5 mg piperine or matched placebo. A general questionnaire, consent form as well as NUTRIC, SOFA and APACHEII scoring system and anthropometrics will be assessed at baseline. The inflammatory markers including TNF-α, IL-6, MCP-1 and CRP, oxidative stress indices (GPx and SOD) and adipokines (leptin and adiponetctin) will be measured at baseline and at the end of the study. In addition, dietary intake, concomitant drugs and laboratory tests will be recorded daily. DISCUSSION: To the best of our knowledge, this is the first clinical trial investigating the effect of curcuminoids supplementation in critically ill patient with TBI. The findings of the present study will provide evidence on the efficacy and safety of curcuminoids in these patients. TRIAL REGISTRATION NUMBER: (http://www.irct.ir, identifier: IRCT20180619040151N1), Registration date:18.09.2018.

Strategies for Prevention and Treatment of Peritoneal Fibrosis: A Scientometric Study.

BACKGROUND: Interest in using peritoneal dialysis (PD) shows global and national increase. However, it remains a challenge to prevent the progression of PD-associated fibrosis in clinical practice. Here, we assessed the status of scientific publications in prevention and management of PD-associated fibrosis in a scientometric study. METHODS: We retrieved the bibliometric data by search terms "encapsulating peritoneal fibrosis," "treatment or prevention," and their synonyms in the Scopus databases until December 2, 2017. Data were analyzed using Scopus analysis tools, SPSS version 15 and Visualizing Scientific Landscapes viewer version 1.6.5. RESULTS: Number of publications showed a steady significant increase (P < 0.001) reaching to 390 documents. Japan had the highest share (21.3%) followed by United Kingdom. Coauthorship network assessment assigned "Ikeda M." from Japan as the top author. The top source of documents was "Peritoneal Dialysis International." Most of documents were original articles focusing on prevention and management of malignant fibrosis of peritoneum (72.6%). The documents were cited totally 5636 times with average citations per article of 14.45, and relatively high H-index of 38. CONCLUSIONS: Despite the global increasing trend in scientific output in this field, contribution of our country is very small. Perhaps more national and international collaboration is required to encourage our researchers for producing more scientific products.

Clinical and Bacterial Risk Factors for Development of Post-Prostate Biopsy Infections.

PURPOSE: To research on clinical and bacterial risk factors and their relationship with post-prostate biopsy infection (PBI). MATERIALS AND METHODS: In this prospective cohort study, rectal swabs were collected from 158 men prior to prostate biopsy and cultured selectively for identify ciprofloxacin-resistant (FQ-R) gram-negative bacteria. The patient characteristics, phylogenetic background, sequence typing and pulsed field gel electrophoresis (PFGE) pattern were compared in two groups of FQ-R E. coli rectal and clinical isolates. RESULTS: In total, PBI was observed in 20 (12.5%) cases; the most of these subjects were FQ-R-colonized. (17/73 [24%] vs 3/85 [3.5%]; P < 0.001). FQ-R colonization, diabetes, hospitalization and UTI were independent risk factors (95% CI: 1.1-20.1, OR = 4.73; 95% CI: 1.7-25.3, OR = 6.57; 95% CI: 1.9-27.5, OR = 7.22; and 95% CI: 1.2-14.3, OR = 4.05; respectively),  that increased the rate of PBI (All P < 0.05). Despite the increase in infections among patients colonized with strains of E. coli ST131, its prevalence was near significance between colonized and infected groups (P = 0.07). The PFGE patterns and antimicrobial susceptibility profiles of rectal and clinical isolates in 13 patients were similar which is remarkably important and informative. CONCLUSIONS: The most PBIs originate from FQ-R E. coli rectal colonization. Rectal culture screening and assessment of clinical risk factors can predict the incidence of PBI in patients.

Systemic Infusion of Autologous Adipose Tissue-Derived Mesenchymal Stem Cells in Peritoneal Dialysis Patients: Feasibility and Safety.

OBJECTIVE: Using mesenchymal stem cells (MSCs) is regarded as a new therapeutic approach for improving fibrotic diseases. the aim of this study to evaluate the feasibility and safety of systemic infusion of autologous adipose tissue-derived MSCs (AD-MSCs) in peritoneal dialysis (PD) patients with expected peritoneal fibrosis. MATERIALS AND METHODS: This study was a prospective, open-label, non-randomized, placebo-free, phase I clinical trial. Case group consisted of nine eligible renal failure patients with more than two years of history of being on PD. Autologous AD-MSCs were obtained through lipoaspiration and expanded under good manufacturing practice conditions. Patients received 1.2 ± 0.1×106 cell/kg of AD-MSCs via cubital vein and then were followed for six months at time points of baseline, and then 3 weeks, 6 weeks, 12 weeks, 16 weeks and 24 weeks after infusion. Clinical, biochemical and peritoneal equilibration test (PET) were performed to assess the safety and probable change in peritoneal solute transport parameters. RESULTS: No serious adverse events and no catheter-related complications were found in the participants. 14 minor reported adverse events were self-limited or subsided after supportive treatment. One patient developed an episode of peritonitis and another patient experienced exit site infection, which did not appear to be related to the procedure. A significant decrease in the rate of solute transport across peritoneal membrane was detected by PET (D/P cr=0.77 vs. 0.73, P=0.02). CONCLUSION: This study, for the first time, showed the feasibility and safety of AD-MSCs in PD patients and the potentials for positive changes in solute transport. Further studies with larger samples, longer follow-up, and randomized blind control groups to elucidate the most effective route, frequency and dose of MSCs administration, are necessary (Registration Number: IRCT2015052415841N2).

In Vitro Spermatogenesis by Three-dimensional Culture of Spermatogonial Stem Cells on Decellularized Testicular Matrix.

BACKGROUND: In the males, Spermatogonial Stem Cells (SSCs) contribute to the production of sex cells and fertility. In vitro SSCs culture can operate as an effective strategy for studies on spermatogenesis and male infertility treatment. Cell culture in a three-dimensional (3D) substrate, relative to a two-dimensional substrate (2D), creates better conditions for cell interaction and is closer to in vivo conditions. In the present study, in order to create a 3D matrix substrate, decellularized testicular matrix (DTM) was used to engender optimal conditions for SSCs culture and differentiation. MATERIALS AND METHODS: After, testicular cells enzymatic extraction from testes of brain-dead donors, the SSCs were proliferated in a specific culture medium for four weeks, and after confirming the identity of the colonies derived from the growth of these cells, they were cultured on a layer of DTM as well as in 2D condition with a differentiated culture medium. In the Sixth week since the initiation of the differentiation culture, the expression of pre meiotic (OCT4 & PLZF ), meiotic (SCP3 & BOULE) and post meiotic (CREM & Protamine-2) genes were measured in both groups. RESULTS: The results indicated that the expression of pre meiotic, meiotic and post meiotic genes was significantly higher in the cells cultured on DTM (P ≤ 0.001). CONCLUSION: SSCs culture in DTM with the creation of ECM and similar conditions with in vivo can be regarded as a way of demonstrating spermatogenesis in vitro, which can be adopted as a treatment modality for male infertility.

Efficiency of colony formation and differentiation of human spermatogenic cells in two different culture systems.

Optimization of in vitro culture system for the expansion and the maturation of male germ cells to post meiotic stages is a valuable tool for studies exploring spermatogenesis regulation and the management of male infertility. Several studies have reported promising results of mouse spermatogonial stem cells culture in three-dimensional (3D) culture systems and a subsequent production of sperm. In the present study, we investigated the capacity of a three-dimensional soft agar culture system (SACS) supplemented with Knockout Serum Replacement (KSR) in colony formation and inducing human germ cells to reach post-meiotic stages. Testicular cells from testes of brain -dead donors were first cultured for three weeks in proliferation medium. The cells were subsequently cultured in the upper layer of the SACS (3D group) in a medium supplemented with KSR and hormones, and the results were compared with that of a two-dimensional (2D) culture system. We found that the number and diameter of colonies and the levels of expression of Scp3 and Integrin α6 in the 3D culture group were significantly higher than in the 2D group. Our findings indicate that SACS can reconstruct a microenvironment capable of regulating both proliferation and differentiation of cell colonies.

Monitoring cellular immune function of renal transplant recipients based on adenosine triphosphate (ATP) production by mitogen-induced CD4+ T helper cells.

Maintaining the balance between over- and under-immunosuppression has a critical role for successful immunosuppressive therapy after renal transplantation. We studied the predictive value of our functional immune assay, which works based on adenosine triphosphate (ATP) levels, in determining risk of infection and rejection among renal transplant recipients (RTRs). A total of 65 RTRs with less than 1 month (RTRL1) and 48 RTRs with more than 6 months (RTRM6) of post-transplant time, and 56 healthy individuals were included. Upon lymphocyte activation by phytohemagglutinin (PHA), CD4+ T cells were separated using magnetic beads (Dynabeads), the intracellular ATP (iATP) concentrations were measured by luciferin-luciferase reaction, and compared within and between the groups. Activated CD4+ cells iATP production directly correlated with post-transplant time (r = 0.32, P = 0.011). The iATP levels were significantly lower in both RTRL1 and RTRM6 groups compared to control (P < 0.001), and in the RTRL1 group compared to the RTRM6 (P < 0.05). The iATP concentrations were significantly lower in patients who suffered from infection versus the RTRs with stable graft function (SGF). However, the iATP levels were higher in those with allograft rejection episode (ARE). Our optimization experiments showed that best iATP levels cutoffs were 472.5 and 572.5 ng/ml for predicting risk of ARE, and 218.5 and 300.5 ng/ml for predicting risk of developing infection in RTRL1 and RTRM6 patients, respectively. iATP levels measured by immune function assay might be a promising predictive tool for identifying RTRs who are at risk of developing infection or allograft rejection.

Organ culture of seminiferous tubules using a modified soft agar culture system.

BACKGROUND: In-vitro spermatogenesis in mammalian species is considered an important topic in reproductive biology. New strategies for achieving a complete version of spermatogenesis ex vivo have been conducted using an organ culture method or culture of testicular cells in a three-dimensional soft agar culture system (SACS). The aim of this study was to develop a new method that supports spermatogenesis to the meiotic phase and morphologically mature spermatozoa through the culture of testicular cells and seminiferous tubules (STs) in a modified SACS, respectively. METHODS: First, enzymatically dissociated testicular cells and mechanically dissociated STs of neonatal mice were separately embedded in agarose and then placed on the flat surface of agarose gel half-soaked in the medium to continue culture with a gas-liquid interphase method. RESULTS: Following 40 days of culture, the meiotic (Scp3) and post-meiotic (Acr) gene expression in aggregates and STs was confirmed by real-time polymerase chain reaction. These results were complemented by immunohistochemistry. The presence of morphologically mature spermatozoa in the frozen sections of STs was demonstrated with hematoxylin and eosin staining. We observed Plzf- or Integrin α6-positive spermatogonia in both cultures after 40 days, indicating the potency of the culture system for both self-renewal and differentiation. CONCLUSIONS: This technique can be used as a valuable approach for performing research on spermatogenesis and translating it into the human clinical setting.