The effect of kisspeptin on the maturation of human ovarian follicles in culture following vitrification-thawing processes.

OBJECTIVE: Ovarian cryopreservation is one of the effective methods to preserve fertility for cancer patients. Still, this approach has some problems, namely ROS, resulting in adverse effects on oocytes and ovarian follicles. Kisspeptin as an antioxidant to control ovarian function, directly or indirectly. In this study, the effect of kisspeptin on follicle maturation was evaluated in culture following ovarian cryopreservation. METHODS: Ovarian tissue samples of women between 20 and 35 years old (n=12) were laparoscopically collected. The samples were randomly divided into four groups: 1) control, 2) vitrification, 3) vitrified+1µM kisspeptin, and 4) vitrified+10µM kisspeptin. After vitrification and thawing processes, the tissues were cultured in DMEM medium for 7 days. H&E staining for histological evaluation, Real-Time PCR for GDF9 and BMP15 gene expression, and immunohistochemical staining for GDF9 and BMP15 protein expression were performed. RESULTS: In the vitrification group, ovarian tissue morphology was incoherent, and more primordial follicles than other follicle types were found. The expression of GDF9 and BMP15 genes and proteins were significantly decreased in this group compared with other groups (p<0.05). In the vitrification groups with kisspeptin (1 and 10 µM), the number of primary and secondary follicles was more than in the vitrification group. Besides, the expression of these genes and proteins was dramatically elevated in the vitrification groups with kisspeptin compared to the vitrification group alone (p<0.05). CONCLUSIONS: It seems that kisspeptin is an effective substance to improve the quality of the human ovarian cryopreservation medium by improving follicle maturation.

The EDA-deficient mouse has Zymbal's gland hypoplasia and acute otitis externa.

In mice, rats, dogs and humans, the growth and function of sebaceous glands and eyelid Meibomian glands depend on the ectodysplasin signalling pathway. Mutation of genes encoding the ligand EDA, its transmembrane receptor EDAR and the intracellular signal transducer EDARADD leads to hypohidrotic ectodermal dysplasia, characterised by impaired development of teeth and hair, as well as cutaneous glands. The rodent ear canal has a large auditory sebaceous gland, the Zymbal's gland, the function of which in the health of the ear canal has not been determined. We report that EDA-deficient mice, EDAR-deficient mice and EDARADD-deficient rats have Zymbal's gland hypoplasia. EdaTa mice have 25% prevalence of otitis externa at postnatal day 21 and treatment with agonist anti-EDAR antibodies rescues Zymbal's glands. The aetiopathogenesis of otitis externa involves infection with Gram-positive cocci, and dosing pregnant and lactating EdaTa females and pups with enrofloxacin reduces the prevalence of otitis externa. We infer that the deficit of sebum is the principal factor in predisposition to bacterial infection, and the EdaTa mouse is a potentially useful microbial challenge model for human acute otitis externa.

A novel transgenic mouse strain expressing PKCßII demonstrates expansion of B1 and marginal zone B cell populations.

Protein kinase Cß (PKCß) expressed in mammalian cells as two splice variants, PKCßI and PKCßII, functions in the B cell receptor (BCR) signaling pathway and contributes to B cell development. We investigated the relative role of PKCßII in B cells by generating transgenic mice where expression of the transgene is directed to these cells using the Eµ promoter (Eµ-PKCßIItg). Our findings demonstrate that homozygous Eµ-PKCßIItg mice displayed a shift from IgD+IgMdim toward IgDdimIgM+ B cell populations in spleen, peritoneum and peripheral blood. Closer examination of these tissues revealed respective expansion of marginal zone (MZ)-like B cells (IgD+IgM+CD43negCD21+CD24+), increased populations of B-1 cells (B220+IgDdimIgM+CD43+CD24+CD5+), and higher numbers of immature B cells (IgDdimIgMdimCD21neg) at the expense of mature B cells (IgD+IgM+CD21+). Therefore, the overexpression of PKCßII, which is a phenotypic feature of chronic lymphocytic leukaemia cells, can skew B cell development in mice, most likely as a result of a regulatory influence on BCR signaling.

The Effect Of Preoperative Gabapentin on Pain Severity After Posterior Urethral Surgery: A Randomized, Double-Blind, Placebo-Controlled Study.

PURPOSE: Prevention and treatment of urethroplastic postoperative pain is a major challenge in post-surgery care. gabapentin can treat postoperative pain by preventing excessive sensitivity to the central nervous system. we have tried to compare the effect of gabapentin with the placebo on reducing the pain associated with posterior urethroplasty in patients. MATERIAL AND METHOD: This prospective, randomized, double-blind study was scheduled in Shohad e Tajrish hospital where a single dose of Gabapentin was compared with placebo given to patients preoperatively. In this study,100 patients with posterior urethral stricture were included for surgery with 50 patients in each arms .All patients underwent posterior urethroplasty. After surgery, the pain level is assessed and evaluated by the visual analog scale in two hours,four hours, six hours,eight hours, twelve hours, and 24 hours after the operation. RESULT: In this study, there was a significant difference in the pain level that was evaluated by the visual analog scale in two hours, four hours, six hours, eight hours, twelve hours, and 24 hours after the surgery (p.value <0.001). We also found a significant decrease in morphine consumption in the gabapentin group vs the placebo group. (p.value <0.001) The post-surgery assessments showed significant lower adverse effects such as vomiting, nausea, drowsiness, and pruritus in the gabapentin group vs. placebo group. CONCLUSION: The consequences of this study revealed that gabapentin effective in controlling posterior urethroplasty postoperative pain, consumption of opioid, nausea, vomiting, drowsiness, and pruritus compared with the placebo group.

End to End urethroplasty Post Urethral Uventa Stent Stricture, The First Case Report.

Nowadays there is not any specific technique for repairing the recurrent urethral stricture with retained urethral stent. We report a 49 year-old man with history of end to end urethroplasty 11 years ago who was referred with urethral stricture. He refused to undergo urethroplasty again, so the stricture was managed by Uventa stent insertion which failed after six months. Finally the patient underwent end to end urethroplasty with complete excision of the obstructed urethra, stent and surrounding periurethral fibrosis. End to end urethroplasty post Uventa stent stricture is an available option with good postoperative outcomes.

Persian version of Patient-Reported Outcome Measure for Urethral Stricture Surgery (USS-PROM) Questionnaire, Validation and Adaptation Study.

PURPOSE: The aim of the present study was translation, cross cultural adaptation and face validity evaluation of the Persian version of Patient-Reported Outcome Measure for Urethral Stricture Surgery (USS-PROM) Questionnaire. MATERIALS AND METHODS: This study was assessed: translation, translation quality, reverse translation and comparison of the english version, Content validity, internal consistency and stability. Content validity presents by index of content validity (CVI) and the content validity ratio (CVR). Internal consistency reliability was tested by Cronbach's ?, and test-retest reliability was evaluated by Intraclass Correlation Coefficient (ICC) assessed by Guttman two way mixed absolute agreements. RESULT: Frothy males with history urethroplasty and mean age of 41.4±9.08 (range of 19 to 52) years old; enrolled.  In the case of mean scores of difficulty from the 16 translated items, 80% had easy translation. In terms of translation quality, 92% were the satisfactorily clear. In terms of similar concept, 92% were satisfactory. The overall quality of the translation was satisfactory at 88%. The translated questionnaire has a good internal consistency (Cronbach's alpha as 0.84). CVI and the CVR, test-retest ICC evaluation were appropriate/acceptable in all questions. The questionnaire ICC was .791(CI 95%, .678-.876). Two main different aspects of the questionnaire consisted of urinary symptoms (question 1-10) and Quality of life (question 11-15) Cronbach's alpha were .800 and .671 respectively. CONCLUSION: The Persian version of the questionnaire has acceptable cultural adaptation and face Validity. Further studies should be done using this translated tool to determine its applicability in the urethroplasty patients.

Comparison of Stone Retrieval Basket, Stone Cone and Holmium Laser: Which One Is Better in Retropulsion and Stone-Free Status for Patients with Upper Ureteral Calculi?

Introduction: Transurethral lithotripsy (TUL) is an appropriate treatment for ureteral stones and is usually used for stones in the middle and lower part of the ureter. Different devices such as the Holmium laser, the stone basket, and the stone cone exist to prevent any fragments from retropulsion during TUL. The present study aims to compare the advantages and disadvantages of the Holmium laser, the stone basket, and the stone cone. Methods: A retrospective study was conducted from September 2016 to January 2018 comparing various TUL methods in 88 subjects with proximal ureteral calculi. The study participants were divided into 4 matched groups. The first one included 20 patients undergoing TUL with no device (group 1), the second group included 22 patients undergoing TUL while using the stone retrieval basket, the third group included 18 patients undergoing TUL while utilizing the stone cone and the fourth group included 28 patients undergoing TUL while using the Hol-YAG laser. Results: A residual stone ≥3 mm was recorded in 15.9% of the patients. The stone free rate was seen in 100%, 90.9, 83.3%, and 55% of the Holmium laser group, the retrieval basket group, the stone cone group and the no device group respectively (P=0.001). The lowest rate of surgery complications including ureteral perforation, post-operative fever, and mucosal damage between the 4 groups (P=0.003) and the highest time of surgery (P=0.001) belonged to the laser group. If we want to ignore the laser group, the success rate for lithotripsy was better in both groups with a stone retrieval device compared to the no device group, but no advantage existed between the stone basket and the stone cone. Conclusion: We can safely conclude that lasers significantly help to prevent stone migration during TUL. If we want to ignore the laser group, the success rate for lithotripsy was significantly better in both groups with a stone retrieval device compared to the no device group, but no advantage existed between the stone basket and the stone cone.

Targeting the RNA m6A Reader YTHDF2 Selectively Compromises Cancer Stem Cells in Acute Myeloid Leukemia.

Acute myeloid leukemia (AML) is an aggressive clonal disorder of hematopoietic stem cells (HSCs) and primitive progenitors that blocks their myeloid differentiation, generating self-renewing leukemic stem cells (LSCs). Here, we show that the mRNA m6A reader YTHDF2 is overexpressed in a broad spectrum of human AML and is required for disease initiation as well as propagation in mouse and human AML. YTHDF2 decreases the half-life of diverse m6A transcripts that contribute to the overall integrity of LSC function, including the tumor necrosis factor receptor Tnfrsf2, whose upregulation in Ythdf2-deficient LSCs primes cells for apoptosis. Intriguingly, YTHDF2 is not essential for normal HSC function, with YTHDF2 deficiency actually enhancing HSC activity. Thus, we identify YTHDF2 as a unique therapeutic target whose inhibition selectively targets LSCs while promoting HSC expansion.

Role of ectodysplasin signalling in middle ear and nasal pathology in rat and mouse models of hypohidrotic ectodermal dysplasia.

Patients with mutations in the ectodysplasin receptor signalling pathway genes - the X-linked ligand ectodysplasin-A (EDA), the receptor EDAR or the receptor adapter EDARADD - have hypohidrotic ectodermal dysplasia (HED). In addition to having impaired development of teeth, hair, eccrine sweat glands, and salivary and mammary glands, HED patients have ear, nose and throat disease. The mouse strains Tabby (EdaTa ) and downless (Edardl-J/dl-J ) have rhinitis and otitis media due to loss of submucosal glands in the upper airway. We report that prenatal correction of EDAR signalling in EdaTa mice with the agonist anti-EDAR antibody rescues the auditory-tube submucosal glands and prevents otitis media, rhinitis and nasopharyngitis. The sparse- and wavy-haired (swh) rat strain carries a mutation in the Edaradd gene and has similar cutaneous HED phenotypes to mouse models. We report that auditory-tube submucosal glands are smaller in the homozygous mutant Edaraddswh/swh than those in unaffected heterozygous Edaraddswh/+ rats, and that this predisposes them to otitis media. Furthermore, the pathogenesis of otitis media in the rat HED model differs from that in mice, as otitis media is the primary pathology, and rhinitis is a later-onset phenotype. These findings in rodent HED models imply that hypomorphic as well as null mutations in EDAR signalling pathway genes may predispose to otitis media in humans. In addition, this work suggests that the recent successful prenatal treatment of X-linked HED (XLHED) in humans may also prevent ear, nose and throat disease, and provides diagnostic criteria that distinguish HED-associated otitis media from chronic otitis media with effusion, which is common in children.

Chronic otitis media is initiated by a bulla cavitation defect in the FBXO11 mouse model.

Auditory bulla cavitation defects are a cause of otitis media, but the normal cellular pattern of bulla mesenchyme regression and its failure are not well understood. In mice, neural-crest-derived mesenchyme occupies the bulla from embryonic day 17.5 (E17.5) to postnatal day 11 (P11) and then regresses to form the adult air-filled bulla cavity. We report that bulla mesenchyme is bordered by a single layer of non-ciliated epithelium characterized by interdigitating cells with desmosome cell junctions and a basal lamina, and by Bpifa1 gene expression and laminin staining of the basal lamina. At P11-P12, the mesenchyme shrinks: mesenchyme-associated epithelium shortens, and mesenchymal cells and extracellular matrix collagen fibrils condense, culminating in the formation of cochlea promontory mucosa bordered by compact non-ciliated epithelial cells. FBXO11 is a candidate disease gene in human chronic otitis media with effusion and we report that a bulla cavitation defect initiates the pathogenesis of otitis media in the established mouse model Jeff (Fbxo11Jf/+ ). Persistent mesenchyme in Fbxo11Jf/+ bullae has limited mesenchymal cell condensation, fibrosis and hyperplasia of the mesenchyme-associated epithelium. Subsequent modification forms fibrous adhesions that link the mucosa and the tympanic membrane, and this is accompanied by dystrophic mineralization and accumulation of serous effusion in the bulla cavity. Mouse models of bulla cavitation defects are important because their study in humans is limited to post-mortem samples. This work indicates new diagnostic criteria for this otitis media aetiology in humans, and the prospects of studying the molecular mechanisms of murine bulla cavitation in organ culture.

Ectodysplasin signalling deficiency in mouse models of hypohidrotic ectodermal dysplasia leads to middle ear and nasal pathology.

Hypohidrotic ectodermal dysplasia (HED) results from mutation of the EDA, EDAR or EDARADD genes and is characterized by reduced or absent eccrine sweat glands, hair follicles and teeth, and defective formation of salivary, mammary and craniofacial glands. Mouse models with HED also carry Eda, Edar or Edaradd mutations and have defects that map to the same structures. Patients with HED have ear, nose and throat disease, but this has not been investigated in mice bearing comparable genetic mutations. We report that otitis media, rhinitis and nasopharyngitis occur at high frequency in Eda and Edar mutant mice and explore the pathogenic mechanisms related to glandular function, microbial and immune parameters in these lines. Nasopharynx auditory tube glands fail to develop in HED mutant mice and the functional implications include loss of lysozyme secretion, reduced mucociliary clearance and overgrowth of nasal commensal bacteria accompanied by neutrophil exudation. Heavy nasopharynx foreign body load and loss of gland protection alters the auditory tube gating function and the auditory tubes can become pathologically dilated. Accumulation of large foreign body particles in the bulla stimulates granuloma formation. Analysis of immune cell populations and myeloid cell function shows no evidence of overt immune deficiency in HED mutant mice. Our findings using HED mutant mice as a model for the human condition support the idea that ear and nose pathology in HED patients arises as a result of nasal and nasopharyngeal gland deficits, reduced mucociliary clearance and impaired auditory tube gating function underlies the pathological sequelae in the bulla.

Preventive effect of tamsulosin on postoperative urinary retention.

PURPOSE: To investigate the prophylactic effect of Tamsulosin, a super-selective alpha-1a adrenergic blocking agent, on the development of urinary retention in men undergoing elective inguinal herniorrhaphy. MATERIALS AND METHODS: From May 2010 through November 2011, a total of 80 males who underwent elective inguinal herniorrhaphy in a university hospital were included in this study. Patients were randomly assigned to one of two groups. In group one (control), the patients were given two doses of placebo orally, 6 hours before surgery and 6 to 12 hours after surgery. Patients in group two were given 0.4 mg of Tamsulosin orally in the same manner as the placebo. All patients were closely followed for 24 hours post-operatively, and any voiding difficulties or urinary retention was recorded. RESULTS: There were 40 patients in group one (control group) and 40 patients in group two (Tamsulosin group). The patients' mean age was 64 years. In group one, 6 patients and in group two, 1 patient required catheterization. Thus, 15% of patients in group I and 2.5% of patients in group II had urinary retention. The difference in the requirement for catheterization was statistically significant (p=0.04). The technique of herniorrhaphy, the side of the body in which the hernia was located, the type of anesthesia, the duration of the surgery, and the severity of pre-operative urinary symptoms had no significant effect on the incidence of urinary retention. CONCLUSIONS: The use of perioperative Tamsulosin represents an effective strategy to reduce the risk of post-operative urinary retention following inguinal herniorrhaphy.

Megacystis secondary to congenital urethral stricture.

Functional bladder capacity at a certain age can be accurately estimated and expressed as a function of age. We present a 13-year-old boy with abdominal distention who was presumed to have ascites. Abdominal ultrasound revealed that the bladder was severely distended, and cystoscopy showed a short stricture in the bulbar urethra followed by a large bladder without obvious borders. Under general anesthesia, bladder capacity was 9250 cc. Reduction cystoplasty was performed. At the 2-year follow-up, he voided spontaneously with 20 cc post-void residual urine.