Expression patterns of sex differentiation-related genes during gonadal sex change in the protogynous wrasse, Halichoeres trimaculatus.

The three-spot wrasse, Halichoeres trimaculatus, can change sex from female to male (i.e. protogyny) due to sharp decrease in endogenous estrogen. During the sex change, ovarian tissue degenerates and testicular tissue arises newly. Finally, ovarian tissue disappears completely and replaces into mature testis. In order to predict the molecular mechanisms controlling the processes of sex change, we investigated the expression patterns of four genes (rspo1, figla, sox9b and amh), which have been thought to be associated with ovarian/testicular differentiation in vertebrates. Expression levels of rspo1 and figla, which play important roles for ovarian differentiation in vertebrates, were stable until the middle stage of the sex change, and subsequently down-regulated. Therefore, it was indicated that decrease in rspo1 and figla could result from ovarian degeneration. On the other hand, basis on the expression pattern, it was indicated that sox9b and amh, which are involved in testicular differentiation in vertebrates, were implicated in testicular formation and spermatogenesis during the sex change as well. The present results could be fundamental information for investigating the relationship between these factors and E2 depletion, which is crucial trigger for sex change.

Relationship between Vertebral Artery Hypoplasia and Posterior Circulation Ischemia.

PURPOSE: Vertebral artery hypoplasia (VAH) is a common congenital anatomical variation. In previous reports, it was unclear whether VAH was an independent risk factor for posterior circulation ischemia. The purpose of this study was to evaluate the impact of VAH on posterior circulation ischemia. METHODS: Subjects were patients with acute ischemic stroke who underwent brain magnetic resonance imaging (MRI) and carotid ultrasonography. Diagnostic criteria for VAH were as follows: (1) Vertebral artery (VA) diameter less than 2.5 mm; (2) VA diameter less than 3.0 mm and a difference in length equal to or greater than 1:1.7; (3) VA diameter less than 3.0 mm, peak systolic velocity less than 40 cm/second, and resistance index value greater than .75. The patients were categorized by the location of the ischemic stroke on MRI as follows: lesion in posterior circulation (P group), lesion in anterior circulation (A group), and multiple lesions in both the anterior and posterior circulations (AP group). RESULTS: We evaluated 129 consecutive patients. VAH was seen in 39, and VA occlusion was found in 15. The prevalence of VAH in the P group (44.4%) was significantly higher than that in the A + AP group (24.7%, P = .034). Multivariate regression analysis showed that large-artery atherosclerosis (odds ratio, 6.3; 95% confidence interval [CI], 1.3-30.1), posterior circulation ischemia (odds ratio, 12.0; 95% CI, 2.8-51.2), and VAH (odds ratio, 4.2; 95% CI, 1.2-15.0) were independent factors related to VA occlusion. CONCLUSION: VAH was an independent factor related to VA occlusion. Therefore, VAH likely plays a role in posterior circulation ischemia.

Clinical Characteristics of Intracranial Reversed Vertebral Artery Flow Evaluated by Transcranial Color Flow Imaging.

BACKGROUND: Carotid duplex ultrasonography (CUS) has been used to identify reversed vertebral artery flow (RVAF) at the extracranial cervical artery in some patients with subclavian steal syndrome. However, the characteristics of intracranial RVAF as evaluated by transcranial color flow imaging (TC-CFI), which can examine intracranial hemodynamics in a real-time and noninvasive fashion, remain unclear. The goal of this study was to analyze the prevalence of intracranial RVAF and its associated clinical characteristics. METHODS: Subjects were consecutive patients who underwent TC-CFI and CUS. We evaluated blood flow in both intracranial vertebral arteries (VAs) from the suboccipital echo window using TC-CFI. RVAF was defined as a flow signal directed toward the probe. We calculated the prevalence of intracranial RVAF in our subjects. Then, we investigated vascular condition (ie, site of lesion, stenosis, occlusion, and dissection) using magnetic resonance angiography, computed tomography angiography , and CUS in patients with intracranial RVAF. RESULTS: Seven hundred twenty patients (508 men; median age, 73 years) were included in this study from September 2007 to March 2013. Intracranial RVAF was seen in 12 patients (1.7%; 11 men; median age, 61 years). Among the 12 patients with intracranial RVAF, 8 patients (67%) had ischemia of the vertebrobasilar territory with distal VA occlusion, according to CUS. Of those patients, 6 (75%) had dissection of the VA. CONCLUSIONS: TC-CFI detected intracranial RVAF in 1.7% of consecutive examinations in our facility. In vertebrobasilar territory stroke patients with intracranial RVAF, VA dissection may contribute to the development of stroke.

[A Case of Tubercular Prostatic Abscess Following Intravesical Bacillus Calmette-Guerin Therapy].

We report a case of tubercular prostatic abscess. A 65-year-old man had undergone intravesical Bacillus Calmette-Guerin therapy for a non-muscle invasive bladder carcinoma. One year 8 months later, the prostate-specific antigen concentration in serum was elevated (18. 58 ng/ml). Results of magnetic resonance imaging (MRI) of the pelvis led us to suspect a prostatic abscess, and transurethral resection of the prostate for drainage was performed. A culture of fluid obtained from the latter procedure revealed a tubercular prostatic abscess. We administered the antituberculous agents, isoniazid (300 mg) and rifampicin(450 mg) daily, for 6 months. One year after surgery, the patient had no urinary symptoms or evidence of recurrence.

Characterization of gonadal soma-derived factor expression during sex change in the protogynous wrasse, Halichoeres trimaculatus.

BACKGROUND: Sex change in fishes provides a good experimental model for understanding the mechanisms and plasticity of sex determination and differentiation. The three-spot wrasse, Halichoeres trimaculatus is a protogynous hermaphrodite. During sex change from female to male, the ovary is replaced by the testis through the degeneration of oocytes and subsequent spermatogenesis. In the present study, we cloned a cDNA-encoding gonadal soma-derived factor (GSDF) from protogynous wrasse and examined its expression pattern in the sexually mature gonads and the sex-changing gonad induced experimentally by aromatase inhibition. RESULTS: Expression of gsdf was predominantly observed in the testis, and it was mainly localized to the supporting cells surrounding the spermatogonia. In the ovary, only slight expression of gsdf was observed in morphologically undifferentiated supporting cells in contact with oogonia. During sex change, strong expression of gsdf appeared first in the supporting cells surrounding the gonial germ cells before the onset of spermatogenesis. Thereafter, the expression of gsdf continually increased in the supporting cells surrounding the proliferating spermatogonia throughout the sex change. CONCLUSIONS: These results suggest that gsdf is involved in the proliferation of spermatogonia and subsequent spermatogenesis in both the testis and the gonad in the early stages of sex change.

[Sertoli cell tumor of the testis : a case report and questionnaire survey on the reported cases in Japan].

We report a case of Sertoli cell tumor of the testis. A 33-year-old man visited our hospital with the complaints of macroscopic hematuria and fever. The left testis was swollen on palpation. Serum levels of human chorionic gonadotropin-ß and lactate dehydrogenase were not elevated, while α fetoprotein was slightly over the normal range. Ultrasonography showed a hypoechoic lesion in the left testis. There was no evidence of retroperitoneal lymph node enlargement or distant metastasis on computed tomography. A left orchietectomy was performed under the diagnosis of left testicular tumor. The tumor, measuring 20 mm in size was histologically diagnosed as benign Sertoli cell tumor. No adjuvant therapy was performed. Neither recurrence nor evidence of metastasis has been detected for 6 months postoperatively.

[Posterior reversible encephalopathy syndrome (PRES) after kidney transplantation: a case report].

A 60-year-old woman with chronic renal failure due to a polycystic kidney underwent living kidney transplantation. Initial immunosuppressive therapy consisted of tacrolimus (TAC), mycophenolate mofetil (MMF), prednisolone, and basiliximab. Furthermore, rituximab was administered, and double filtration plasmapheresis and plasma exchange were utilized because of ABO-incompatible transplantation, while intravenous immune serum globulin (IVIG) was given because donor specific antibody was positive. Four days after the renal transplantation, the patient developed visual abnormalities, a headache, and paralysis. Then, he became unconscious. Magnetic resonance imaging of the brain demonstrated bilateral posterior vasogenic edema. Our diagnosis was posterior reversible encephalopathy syndrome due to TAC neurotoxicity. After converting TAC to reduced cyclosporine and everolimus, the symptoms rapidly disappeared.

[A case of urachal carcinoma with multiple lung metastases treated by TS-1/CDDP chemotherapy].

A 67-year-old woman presented with macroscopic hematuria and lower abdominal pain. Cystoscopy revealed a broad-stalk non-papillary tumor at the bladder dome. Computed tomography (CT) showed a tumor extending from the umbilicus to the bladder dome, together with multiple lung metastases. Serum carcinoembryonic antigen and cancer antigen (CA19-9) levels were elevated at 7.0 ng/ml and 180 U/ml, respectively. Transurethral resection of the tumor was performed and histopathology revealed adenocarcinoma. Therefore, the tumor was diagnosed as a stage IVB (Sheldon's category) urachal carcinoma. En bloc segmental resection of the urachal carcinoma with the bladder dome was performed, followed by chemotherapy with tegafur, gimestat, and otastat potassium (TS-1) and cisplatin. The disease remained stable for 8 months. However, a follow up CT scan after 11 chemotherapy cycles showed progression of the lung metastases. In spite of the change to a second-line gemcitabine and cisplatin chemotherapy regimen, the disease continued to progress after 4 cycles.

[Complete atrioventricular block due to cardiac metastasis from bladder cancer: case report].

A 72-year-old woman was diagnosed with bladder cancer (cT3bN0M0). After 2 cycles of GC (gemcitabine，cisplatin) neoadjuvant chemotherapy, the patient underwent a cystectomy and ileal conduit construction. Pathological findings showed urothelial carcinoma, high grade, G2＞G3, pT3a, INFß, ly0, v1. Six months after the operation, metastases to the liver, lung, left adrenal gland, rib, multiple lymph nodes, and peritoneum were revealed. Under palliative care, she suffered from palpitation and general fatigue. Electrocardiogram findings showed a complete atrioventricular block, while echocardiography and computed tomography revealed cardiac metastasis. We diagnosed her with complete atrioventricular block due to cardiac metastasis from bladder cancer. She died 7 days after onset of the complete atrioventricular block without use of a pacemaker.

Immunohistochemical features of BK virus nephropathy in renal transplant recipients.

BK virus nephropathy (BKVN) is one of the factors that reduces renal graft function after transplantation. However, BKVN and rejection present similar pathological findings, as both are accompanied by cellular infiltration to the interstitium and tubulitis, thus they are difficult to distinguish for diagnosis and medical treatment. In the present study, we examined immunohistochemical pathological features of BKVN in four cases treated in our hospital from 2007 to 2010. Common immunohistological finding is that tubulitis in these cases was primarily EMA and 34ßE12-positive and existed predominantly from the collecting duct to the distal convoluted tubule. The majority of infected cells existed in EMA and 34ßE12-positive tubules, which were also located mainly from the collecting duct to the distal tubule. In addition, a large number of SV40-positive infected cells were similarly seen. Dylon staining clearly revealed eosinophils. We concluded that the main pathological features of BKVN are the presence of tubulitis and infected cells predominantly from the collecting duct to distal tubule and the appearance of eosinophils.

[Dopamine-secreting pheochromocytoma with neurofibromatosis type 1 : a case report].

Pheochromocytoma occurs in 0.1-5.7% of patients with neurofibromatosis type 1 (NF1), while dopamine-secreting pheochromocytoma is rare. We report here a rare case of dopamine-secreting pheochromocytoma in a patient with NF1. A 46-year-old woman with NF1 was referred to our hospital with epigastralgia. The patient had no history of hypertension. Abdominal sonography incidentally revealed a left adrenal tumor, while abdominal computed tomography and magnetic resonance imaging findings showed a left adrenal tumor, sized 63×58 mm. Laboratory evaluations revealed exclusively elevated urine dopamine levels in addition to elevated serum adrenaline and noradrenaline levels. Laparoscopic left adrenalectomy was performed and the dopamine levels and other cathecolamine levels returned to normal postoperatively.

[A case of malignant peritoneal mesothelioma presenting with bilateral swelling of spermatic cord].

A 67-year-old man had pain and swelling in the bilateral inguinal region. His past medical history included non-alcoholic steatohepatitis and liver cirrhosis. A clinical examination revealed a solid mass that was palpable along with the bilateral spermatic cord from the external inguinal ring to the root of the scrotum. Other than a hydrocele in the right testis, the epididymis and testes were intact. Abdominal computed tomography showed ascites and a solid tumor of the bilateral spermatic cord. Magnetic resonance imaging findings did not suggest malignancy, while antibiotics showed no effects. The patient wanted relief from the inguinal pain and a bilateral high orchiectomy was performed, during which time the spermatic cord was found firmly adhered to the surrounding tissue, and could not be completely resected. A histopathological examination showed epithelial mesothelioma. For additional examination of the ascites, the patient underwent paracentesis, which revealed an extremely high level of hyaluronic acid. Our working diagnosis was peritoneal malignant mesothelioma that had infiltrated the bilateral spermatic cord. We began systemic chemotherapy treatment with pemetrexed and cisplatin, but, the patient died 3 months after the operation.

Roles of Gonadotropin Receptors in Sexual Development of Medaka.

The gonadotropins, follicle-stimulating hormone (FSH) and luteinizing hormone (LH), are secreted from the pituitary and bind to the FSH receptor (FSHR) and LH receptor (LHR) to regulate gonadal development in vertebrates. Previously, using fshr-knockout (KO) medaka (Oryzias latipes), we demonstrated that FSH regulates ovarian development by elevating estrogen levels. However, the lhr-KO phenotype in medaka is poorly characterized. Here, we generated lhr-KO medaka using the transcription activator-like effector nuclease (TALEN) technique. We analyzed its phenotype and that of fshr-KO, lhr;fshr double-heterozygotes (double-hetero), and double-KO fish. All genetically male medaka displayed normal testes and were fertile, whereas fshr-KO and double-KO genetically female fish displayed small ovaries containing many early pre-vitellogenic oocytes and were infertile. Although lhr-KO genetically female fish had normal ovaries with full-grown oocytes, ovulation did not occur. Levels of 17α,20ß-dihydroxy-4-pregnen-3-one, which is required for meiotic maturation of oocytes and sperm maturation in teleost fish, were significantly decreased in all KO female medaka ovaries except for double-heteros. Further, 17ß-estradiol levels in fshr-KO and double-KO ovaries were significantly lower than those in double-heteros. These findings indicate that LH is necessary for oocyte maturation and FSH is necessary for follicle development, but that neither are essential for spermatogenesis in medaka.

Molecular cloning of two gonadotropin receptors and their distinct mRNA expression profiles in daily oogenesis of the wrasse Pseudolabrus sieboldi.

In fish, asynchronous development of ovarian follicles, the simultaneous advance of vitellogenesis and oocyte maturation in one ovary, is a rational reproductive strategy to spawn consecutively in one spawning season. In this study, to clarify the mode of action of follicle stimulating hormone (FSH) and luteinizing hormone (LH) in asynchronous ovarian follicle development in daily egg production, we cloned cDNAs of the follicle-stimulating hormone receptor (FSHR) and luteinizing hormone receptor (LHR) in the bambooleaf wrasse (Pseudolabrus sieboldi), which exhibits clear diurnal spawning rhythms over 1 month. In addition, different developmental stages of ovarian follicles were isolated from whole ovaries at various daily time points on 1 day in the spawning season, and mRNA expression levels of FSHR and LHR were analyzed. Sequence analysis showed distinct differences in the number of putative leucine-rich repeats at the extracellular domain between FSHR and LHR, suggesting a difference in ligand-specificity. Real-time PCR analyses revealed that FSHR mRNA was highly expressed in early yolk-stage follicles but decreased at the end of vitellogenesis. In contrast, the expression of LHR mRNA was maintained at low levels in vitellogenic stage follicles but markedly elevated at the end of the vitellogenic and early migratory nucleus stages, thereafter markedly dropping in the late migratory nucleus stage. The present results suggest that co-regulation of vitellogenesis and oocyte maturation in one ovary is controlled by the stage-distinctive expression levels of FSHR and LHR mRNA in ovarian follicles, and daily switching of sensitivity from FSH to LH is required for daily egg production.

[A case of limbic encephalitis associated with leucine-rich glioma-inactivated 1 antibody].

We report a 55-year-old woman with limbic encephalitis associated with leucine-rich glioma-inactivated 1 (LGI1) antibody. She first developed a generalized seizure, following by consciousness loss. Although anticonvulant was initially effective, she began to present frequently with seizure and memory impairment. After eleven months from onset, she was admitted due to generalized seizure. Neurological examination after recovering from the treatment with anesthetic agent demonstrated disorientation and memory impairment. Cerebrospinal fluid analysis was unremarkable. MR brain FLAIR imaging demonstrated high intensity lesions in the medial parts of the both temporal lobes, suggesting limbic encephalitis. There were no signs of malignant tumor detected on systemic examination. LGI1 antibody was positive in the cerebrospinal fluid and we finally diagnosed this patient as having limbic encephalitis associated with LGI1 antibody. She demonstrated a good response to steroid therapy and was discharged after one month.

High temperature causes masculinization of genetically female medaka by elevation of cortisol.

In poikilothermic vertebrates, sex determination is sometimes influenced by environmental factors such as temperature. However, little is known about the molecular mechanisms underlying environmental sex determination. The medaka (Oryzias latipes) is a teleost fish with an XX/XY sex determination system. Recently, it was reported that XX medaka can be sex-reversed into phenotypic males by high water temperature (HT; 32-34 degrees C) treatment during the sex differentiation period. Here we report that cortisol caused female-to-male sex reversal and that metyrapone (an inhibitor of cortisol synthesis) inhibited HT-induced masculinization of XX medaka. HT treatment caused elevation of whole-body levels of cortisol, while metyrapone suppressed the elevation by HT treatment during sexual differentiation. Moreover, cortisol and 33 degrees C treatments inhibited female-type proliferation of germ cells as well as expression of follicle-stimulating hormone receptor (fshr) mRNA in XX medaka during sexual differentiation. These results strongly suggest that HT induces masculinization of XX medaka by elevation of cortisol level, which, in turn, causes suppression of germ cell proliferation and of fshr mRNA expression.

Isolation, characterization and expression analyses of FSH receptor in protogynous grouper.

Follicle-stimulating hormone (FSH) and its receptor (FSHR) play important roles in spermatogenesis. We cloned and characterized the honeycomb grouper Epinephelus merra FSHR (EmFSHR) to elucidate its role in the protogynous sex change in groupers. Reverse transcription-polymerase chain reaction (RT-PCR) analysis suggested that EmFSHR was expressed exclusively in the gonads. In situ hybridization showed the distribution of EmFSHR in the granulosa cells of previtellogenic oocytes and Leydig cells in the testis. Quantitative reverse transcription PCR (RT-qPCR) analysis of gonadal EmFSHR transcripts during the process of sex change indicated that the lowest levels were found in the female phase before sex change. EmFSHR transcripts increased during the early transitional phase, when oocytes began to degenerate in parallel with the initiation of gonial germ cell differentiation into spermatogonia. A dramatic increase in EmFSHR transcription occurred during the late transitional phase, when the gonad contained numerous proliferating male germ cells and many degenerated oocytes. EmFSHR expression remained high until the transformation from ovary to testis was complete. The data reveal that female to male sex change is associated with the upregulation of EmFSHR transcripts, and that this upregulation may be responsible for the development of testicular tissue and the progression of spermatogenesis. Furthermore, how the upregulation of EmFSHR is controlled in the initiation of sex change remains to be elucidated.

[Combination chemotherapy with gemcitabine and paclitaxel in a hemodialysis patient with metastatic urothelial carcinoma].

A 63-year-old man on hemodialysis for chronic renal failure developed a metastatic urothelial carcinoma and was treated with combination chemotherapies. For first line therapy, gemcitabine (GEM) and carboplatin (CBDCA) were given in combination. CBDCA was administered in a target area under the curve of 5.0 mg x min/ml according to the Calvert formula, followed by infusion of GEM at 1,000 mg/m(2), with hemodialysis started 1 hour after the end of CBDCA administration. Treatment-related adverse effects were severe, with grade 4 thrombocytopenia and neutropenia. Following two cycles of that therapy, partial response (PR) was obtained for lung metastasis, while progressive disease (PD) was noted in the liver. For second line therapy, GEM and paclitaxel (PAC) were given in combination. PAC was administered at 110 mg/m(2) before GEM at 1,000 mg/m(2) with hemodialysis given on the interval days. The adverse effect was grade 3 neutropenia, which was considered acceptable. After two cycles of second line therapy, PR was obtained for both lung and liver metastases, and maintained for 6 months. We concluded that GEM/PAC combination therapy is safe and effective for urothelial carcinoma patients undergoing concurrent hemodialysis.

[A case report of pyogenic spondylodiscitis mimicking spinal metastasis of urothelial carcinoma].

A 75-year-old man was admitted to our hospital with a complaint of fever and backpain. He was diagnosed with acute pyelonephritis, and was administered an antibiotic. However, since there was no improvement in the symptoms, he was admitted to our hospital. Computed tomography revealed a tumor which was in contact with a thoracic vertebra. Because he had had a total cystectomy for bladder cancer in the past, we suspected a spinal metastasis of urothelial carcinoma. However, after magnetic resonance imaging, we finally diagnosed it as pyogenic spondylodiscitis, and his condition improved after administration of antibiotics.

Effect of 1,25-dihydroxyvitamin d on testicular morphology and gene expression in experimental cryptorchid mouse: testis specific cDNA microarray analysis and potential implication in male infertility.

PURPOSE: We evaluated the morphological effect and alterations in gene expression caused by 1,25-dihydroxyvitamin D treatment in the mouse testis undergoing experimental cryptorchidism and subsequent orchiopexy. MATERIALS AND METHODS: The mean modified Johnsen score and testicular weight were estimated after 4 weeks of treatment with a 1,25-dihydroxyvitamin D prodrug. We examined sites of vitamin D receptor and mRNA expression, and 1,25-dihydroxyvitamin D analogue accumulation in the mouse testis. Also, we compared alterations in gene expression in the cryptorchid mouse testis with or without 1,25-dihydroxyvitamin D administration by testis specific cDNA microarray. We confirmed protein synthesis of a candidate among up-regulated genes in primary cultures of Sertoli's cells by Western blotting. RESULTS: Mean +/- SEM Johnsen score and testicular weight were increased by 1,25-dihydroxyvitamin D treatment but not significantly (6.12 +/- 0.33 vs 5.27 +/- 0.4 and 49.3 +/- 3.8 mg vs 42.6 +/- 5.5, p = 0.13 and 0.065, respectively). Vitamin D receptor and its mRNA were positive in Sertoli's cells. The 1,25-dihydroxyvitamin D analogue accumulated mainly in Sertoli's cells. Of 2,483 testis specific genes 19 showed up-regulation by 1,25-dihydroxyvitamin D treatment. Of these genes the regulator of cellular cholesterol homeostasis Abca1 was expressed mainly in Sertoli's cells and influenced male fertility. In primary cultures of Sertoli's cells the synthesis of Abca1 protein was increased by 1,25-dihydroxyvitamin D treatment but not by follicle-stimulating hormone or testosterone treatment. CONCLUSIONS: We noted that 1,25-dihydroxyvitamin D contributes to spermatogenesis by up-regulating certain specific genes in Sertoli's cells. Testis specific cDNA microarray analysis and vitamin D supplementation may have implications for managing male infertility.