The magnitude of gonadotoxicity of chemotherapy drugs on ovarian follicles and granulosa cells varies depending upon the category of the drugs and the type of granulosa cells.

STUDY QUESTION: Do different chemotherapy drugs exert the same magnitude of cytotoxicity on dormant primordial follicles and the growing follicle fraction in the ovary in vivo and on mitotic non-luteinized and non-mitotic luteinized granulosa cells in vitro? SUMMARY ANSWER: Cyclophosphamide (alkylating agent) and cisplatin (alkylating like) impacted both primordial and pre-antral/antral follicles and both mitotic and non-mitotic granulosa cells, whereas the anti-metabolite cancer drug gemcitabine was detrimental only to pre-antral/antral follicles and mitotic non-luteinized granulosa cells. WHAT IS KNOWN ALREADY: It is already known that anti-metabolite cancer drugs are less detrimental to the ovary than alkylating and alkylating like agents, such as cyclophosphamide and cisplatin. This assumption is largely based on the results of clinical reports showing lower rates of amenorrhea in women receiving anti-metabolite agent-based regimens compared with those treated with the protocols containing an alkylating drug or a platinum compound. But a quantitative comparison of gonadotoxicity with a histomorphometric proof of evidence has not been available for many chemotherapy drugs. Therefore, we combined in this study in vivo and in vitro models of human and rat origin that allows a comparative analysis of the impact of different chemotherapy agents on the ovary and granulosa cells using real-time quantitative cell indices, histomorphometry, steroidogenesis assays, and DNA damage and cell death/viability markers. We also aimed to investigate if there is a difference between mitotic and non-mitotic granulosa cells in terms of their sensitivity to the cytotoxic actions of chemotherapy drugs with different mechanisms of action. This issue has not been addressed previously. STUDY DESIGN, SIZE, DURATION: This translational research study involved in vivo analyses of ovaries in rats and in vitro analyses of granulosa cells of human and rat origin. PARTICIPANTS/MATERIALS, SETTING, METHODS: For the in vivo assays, 54 4- to 6-week old Sprague-Dawley young female rats were randomly allocated into four groups of 13 to receive a single IP injection of: saline (control), gemcitabine (200 mg/kg), cisplatin (50 mg/kg) or cyclophosphamide (200 mg/kg). The animals were euthanized 72 h later. Follicle counts and serum AMH levels were compared between the groups. In vitro cytotoxicity studies were performed using mitotic non-luteinized rat (SIGC) and human (COV434, HGrC1) granulosa cells, and non-mitotic luteinized human (HLGC) granulosa cells. The cells were plated at a density of 5000 cells/well using DMEM-F12 culture media supplemented with 10% FBS. Chemotherapy agents were used at their therapeutic blood concentrations. The growth of mitotic granulosa cells was monitored real-time using xCelligence system. Live/dead cell and apoptosis assays were also carried out using intravital Yo-Pro-1 staining and cleaved caspase-3 expression, respectively. Estradiol (E2), progesterone (P) and anti-Mullerian hormone (AMH) levels were assayed with ELISA. MAIN RESULTS AND THE ROLE OF CHANCE: Cyclophosphamide and cisplatin caused massive atresia of both primordials and growing follicles in the rat ovary whereas gemcitabine impacted pre-antral/antral follicles only. Cyclophosphamide and cisplatin induced apoptosis of both mitotic non-luteinized and non-mitotic luteinized granulosa cells in vitro. By contrast, cytotoxicity of gemcitabine was confined to mitotic non-luteinized granulosa cells. LIMITATIONS, REASONS FOR CAUTION: This study tested only three chemotherapeutic agents. The experimental methodology described here could be applied to other drugs for detailed analysis of their ovarian cytotoxicity. WIDER IMPLICATIONS OF THE FINDINGS: These findings indicate that in vivo and in vitro cytotoxic actions of chemotherapy drugs on the ovarian follicles and granulosa cells vary depending upon the their mechanism of action and the nature of the granulosa cells.

Evaluation of Benign Paroxysmal Positional Vertigo in American Football Players.

OBJECTIVES: The aim of this investigation was to evaluate the association between posterior channel benign paroxysmal positional vertigo (BPPV) and trauma that is frequently experienced by American football players. MATERIALS AND METHODS: Participants were classified into the following two groups: (1) a study group consisting of 63 male participants aged 18-30 years who had been playing American football for more than 2 years and (2) a control group consisting of 49 male participants aged 18-27 years with no history of otologic/vestibular disease or acute/chronic trauma. Trauma, age, total duration of playing American football, and weekly training hours of subjects in the study group were analyzed to determine any relationship with BPPV occurrence. We performed otologic, audiologic, and vestibular assessments of pure sound audiometry, tympanometry, tandem walking test with eyes open and eyes closed, Romberg, head shaking, roll, and Dix-Hallpike tests to all participants. RESULTS: A positive correlation between the total years of American football played and posterior channel BPPV frequency was observed in the study group. In addition, increasing weekly hours of training was shown to further increase the risk of BPPV. A total of 16 out of 63 athletes experienced BPPV, whereas none of the participants in the control group experienced BPPV. All participants completed the Vertigo Symptom Scale, which revealed that vertigo did not cause any significant negative impact on their training routine and activities of daily living. CONCLUSION: Our results indicate that the weekly training hours and total years of training with American football increase posterior channel BPPV frequency.

Nasopharyngeal Glial Heterotopia with Intracranial Extension: A Case Report.

Nasopharyngeal glial heterotopia is a mass composed of mature neural tissue occurring outside the central nervous system and is extremely rare. The preoperative diagnosis of such a mass in the head and neck region is challenging. In this study, we report a case of a 16-month-old patient presenting with respiratory distress and snoring caused by nasopharyngeal glial heterotopia. Radiologic imaging and histopathology are obligatory for the definitive diagnosis of glial heterotopia. Preoperative evaluation of an intracranial connection is one of the most essential issues in the presence of pediatric nasopharyngeal masses. The gold standard of treatment is surgical excision. Early recognition and early surgical excision by endoscopic or external approach are crucial to relieve respiratory distress and to maintain healthy growth and development.

Derivation of Neural Stem Cells from Mouse Induced Pluripotent Stem Cells.

Neural stem cells (NSCs) derived from induced pluripotent stem cells offer therapeutic tools for neurodegenerative diseases. This review focuses on embryoid body (EB)-mediated stem cell culture techniques used to derive NSCs from mouse induced pluripotent stem cells (iPSCs). Generation of healthy and stable NSCs from iPSCs heavily depends on standardized in vitro cell culture systems that mimic the embryonic environments utilized during neural development. Specifically, the neural induction and expansion methods after EB formation are described in this review.