Age-related changes in circulating INSL3 concentrations and their associations with ovarian conditions in Japanese Black beef cattle.

Information on circulating levels of insulin-like peptide 3 (INSL3) in female domesticated animals is limited, as their concentrations are significantly lower than in males. The objectives of the present study were to 1) develop a sandwich time-resolved fluorescence immunoassay (TRFIA) with higher detectability to measure blood INSL3 concentrations in female cattle, 2) determine INSL3 concentrations in female cattle among age groups and reproductive conditions, and 3) explore associations between INSL3 levels and ultrasonographic ovarian measurements. Blood was collected repeatedly from Japanese Black beef female calves (n = 12; 0－8 mo), heifers (n = 10; 10－26 mo), and cows (n = 20; 27－200 mo). Blood was taken from the cows (n = 13) at follicular, post-ovulatory, and luteal phases, and from cows with follicular cysts (n = 12). Ultrasonography of ovaries was conducted in the calves (n = 12) and the cows without ovarian diseases (n = 9). The ovarian area, as well as the number and diameters of antral follicles ≥ 2 mm, were determined in each ovary. The proposed method detected a difference in plasma INSL3 between calves (0.01 ng/mL) and heifers (0.18 ng/mL). However, the conventional assay showed similar levels for calves and heifers (1.82 vs 2.07 ng/mL). Plasma INSL3 and testosterone concentrations increased from calves to heifers (P < 0.0001), but only INSL3 rose from heifers to cows (P < 0.0001). INSL3 and testosterone concentrations did not change across the estrus cycle in cows, and the levels of both hormones in follicular cystic cows did not differ from those in the follicular phase. Ovarian area, maximal and average follicular diameters, and total volume of all follicles per animal were higher in cows than calves (P < 0.001). Plasma INSL3 concentrations correlated positively with the total volumes of all follicles in calves (P < 0.05) and cows (P < 0.05), whereas testosterone concentrations did not correlate with ovarian follicular measurements. In conclusion, plasma INSL3 concentrations measured by the proposed sandwich TRFIA showed a clear increase from female calves to cows in beef cattle. These results suggest that circulating levels of INSL3, but not of testosterone, are associated with the total volume of all antral follicles in both ovaries per animal in female cattle.

Anisotropy of the superconducting gap in the iron-based superconductor BaFe2(As(1-x)P(x))2.

We report peculiar momentum-dependent anisotropy in the superconducting gap observed by angle-resolved photoemission spectroscopy in BaFe2(As(1-x)P(x))2 (x = 0.30, Tc = 30 K). Strongly anisotropic gap has been found only in the electron Fermi surface while the gap on the entire hole Fermi surfaces are nearly isotropic. These results are inconsistent with horizontal nodes but are consistent with modified s ± gap with nodal loops. We have shown that the complicated gap modulation can be theoretically reproduced by considering both spin and orbital fluctuations.

Distribution of pituitary adenylate cyclase-activating polypeptide (PACAP) in the human testis and in testicular germ cell tumors.

Pituitary adenylate cyclase-activating peptide (PACAP) is a neuropeptide expressed in the central nervous system and peripheral organs. Previous studies revealed the role and distribution of PACAP in the rodent testis, however, its presence in the human testis and in testicular germ cell tumors is not known. We used RT-PCR and immunohistological observations to investigate whether human testicular tissue and testicular germ cell tumors contain PACAP. The mRNAs for PACAP and its receptors were detected in total RNA extracted from human testes. PACAP immunoreactivity was observed in spermatogonia and spermatids from normal testes. In contrast, diffuse PACAP immunopositivity was observed in seminoma tumor cells, while only faint immunoreactivity was observed in embryonal carcinoma cells. Our data suggest that PACAP may play a role in human spermatogenesis and in testicular germ cell tumor development.

Strongly spin-orbit coupled two-dimensional electron gas emerging near the surface of polar semiconductors.

We investigate the two-dimensional highly spin-polarized electron accumulation layers commonly appearing near the surface of n-type polar semiconductors BiTeX (X=I, Br, and Cl) by angular-resolved photoemission spectroscopy. Because of the polarity and the strong spin-orbit interaction built in the bulk atomic configurations, the quantized conduction-band subbands show giant Rashba-type spin splitting. The characteristic 2D confinement effect is clearly observed also in the valence bands down to the binding energy of 4 eV. The X-dependent Rashba spin-orbit coupling is directly estimated from the observed spin-split subbands, which roughly scales with the inverse of the band-gap size in BiTeX.

Orbital-independent superconducting gaps in iron pnictides.

The origin of superconductivity in the iron pnictides has been attributed to antiferromagnetic spin ordering that occurs in close combination with a structural transition, but there are also proposals that link superconductivity to orbital ordering. We used bulk-sensitive laser angle-resolved photoemission spectroscopy on BaFe(2)(As(0.65)P(0.35))(2) and Ba(0.6)K(0.4)Fe(2)As(2) to elucidate the role of orbital degrees of freedom on the electron-pairing mechanism. In strong contrast to previous studies, an orbital-independent superconducting gap magnitude was found for the hole Fermi surfaces. Our result is not expected from the superconductivity associated with spin fluctuations and nesting, but it could be better explained invoking magnetism-induced interorbital pairing, orbital fluctuations, or a combination of orbital and spin fluctuations. Regardless of the interpretation, our results impose severe constraints on theories of iron pnictides.

Suppression of spermatogenesis in ipsilateral and contralateral testicular tissues in patients with seminoma by human chorionic gonadotropin beta subunit.

OBJECTIVES: The pathologic complexity of the testicular tumor makes it difficult to demonstrate exactly the relationship between the impaired spermatogenesis in patients with a testicular tumor and the serum level of the human chorionic gonadotropin beta subunit (beta-hCG). Therefore, we performed quantitative evaluation of spermatogenesis in ipsilateral and contralateral testicular tissues of seminoma to simplify the relation pathologically and endocrinologically and to demonstrate the exact correlation between spermatogenesis and serum beta-hCG levels. METHODS: Fifty-three biopsy specimens from ipsilateral and contralateral testicular tissues of seminoma were analyzed histologically. The quantitative evaluation of spermatogenesis was performed by the mean Johnsen's score count (MJSC). Beta-hCG expression in seminoma was examined immunohistochemically. Serum beta-hCG, testosterone, estradiol, luteinizing hormone, and follicle-stimulating hormone levels were analyzed before orchiectomy. RESULTS: A significant linear relationship (r = -0.82; P <0.005) was found between the serum level of beta-hCG and the MJSC in contralateral testicular tissues but not in ipsilateral ones, although the suppression of spermatogenesis was observed in both sides without suppression of luteinizing hormone and/or follicle-stimulating hormone production. CONCLUSIONS: A clearcut fall in the MJSC with an associated rise in the serum level of beta-hCG was demonstrated in the contralateral testicular tissues but not in the ipsilateral ones of seminoma. It seems most likely that serum beta-hCG suppresses spermatogenesis in both ipsilateral and contralateral testicular tissues without the suppression occurring through the hypothalamus-pituitary-gonadal system, and also that some less well recognized factors affect spermatogenesis, making the relation between serum beta-hCG and MJSC obscure in ipsilateral testicular tissues.

[Transitional cell carcinoma of the ureter with inverted proliferation accompanied with papillary bladder tumor: a case report].

An 85-year-old female was referred to our hospital with chief complaints of right flank pain and gross hematuria. Ultrasonography demonstrated right hydroureteronephrosis and a thumb head-sized solid mass in the lower third of the right ureter. Cystoscopy revealed papillary tumors near the right ureteral orifice. Under the preoperative diagnosis of right ureteral tumor and bladder tumor, transurethral resection of bladder tumor, right nephroureterectomy and partial cystectomy were performed. The gross specimen of the ureter contained a 5 x 3 x 1 cm, polypoid and smooth-surfaced tumor. The pathological diagnosis of the ureteral tumor was transitional cell carcinoma with inverted proliferation, grade 1 >> grade 2. On the other hand, the bladder tumor was papillary transitional cell carcinoma, grade 1. This is a case in which tumor development showed two different types.

[A case of papillary adenocarcinoma of the prostate which was difficult to diagnose].

We report a case of papillary adenocarcinoma of the prostate found by urethroscopy. An 84-year-old male visited our hospital complaining of initial hematuria in July 1997. No abnormal findings were detected despite repeated urological examinations until endoscopic examination revealed fine papillary tumors in the prostatic urethra along with benign prostatic hyperplasia-like prominent left lobe of the prostate in June 2000. Serum prostate specific antigen (PSA) level was 4.1 ng/ml. He underwent transurethral resection of the urethral tumors and the prominent lobe, which was found to contain packed papillary tumors. Both of these tumors were well differentiated papillary adenocarcinoma most likely originating from the prostate because PSA immunostaining was positive. The prostate was irradiated postoperatively. Papillary adenocarcinoma localized in the prostate is difficult to diagnose preoperatively.

Posttranslational modification of the glycosylation inhibiting factor (GIF) gene product generates bioactive GIF.

Glycosylation inhibiting factor (GIF) and macrophage migration inhibitory factor (MIF) share an identical structure gene. Here we unravel two steps of posttranslational modifications in GIF/MIF molecules in human suppressor T (Ts) cell hybridomas. Peptide mapping and MS analysis of the affinity-purified GIF from the Ts cells revealed that one modification is cysteinylation at Cys-60, and the other is phosphorylation at Ser-91. Cysteinylated GIF, but not the wild-type GIF/MIF, possessed immunosuppressive effects on the in vitro IgE antibody response and had high affinity for GIF receptors on the T helper hybridoma cells. In vitro treatment of wild-type recombinant human GIF/MIF with cystine resulted in preferential cysteinylation of Cys-60 in the molecules. The cysteinylated recombinant human GIF and the Ts hybridoma-derived cysteinylated GIF were comparable both in the affinity for the receptors and in the immunosuppressive activity. Polyclonal antibodies specific for a stretch of the amino acid sequence in alpha2-helix of GIF bound bioactive cysteinylated GIF but failed to bind wild-type GIF/MIF. These results strongly suggest that cysteinylation of Cys-60 and consequent conformational changes in the GIF/MIF molecules are responsible for the generation of GIF bioactivity.

Presence of the 55 kDa glycosylation inhibiting factor in human serum.

An ELISA system for the human glycosylation inhibiting factor (GIF) was established using polyclonal antibodies against highly purified 13 kDa recombinant human GIF, and the concentration of GIF in the sera of healthy donors and patients with various diseases was determined. GIF was detected in the sera of most healthy individuals and its concentration tended to increase with age. It was also found that the serum GIF levels markedly increased in some patients with rheumatoid arthritis or malignant tumors. Analysis of serum samples by SDS-PAGE and immunoblotting revealed a 55 kDa protein that has both the GIF antigenic determinant and the TCR alpha chain determinant. A 13 kDa GIF was not detected in the sera. In view of our previous findings on antigen-specific GIF from murine suppressor T cell hybridomas indicating that the 55 kDa GIF is a post-translationally formed conjugate of a TCR alpha chain with 13 kDa GIF, we suspect that the 55 kDa GIF detected in human sera is a human homologue of the murine 55 kDa GIF.

An essential role of mast cells in the development of airway hyperresponsiveness in a murine asthma model.

Immunization of BALB/c mice with alum-adsorbed OVA, followed by three bronchoprovocations with aerosolized OVA, resulted in the development of airway hyperresponsiveness (AHR) and allergic inflammation in the lung accompanied by severe infiltration of eosinophils into airways. In this murine asthma model, administration of monoclonal anti-IL-5 Ab before each Ag challenge markedly inhibited airway eosinophilia, but the treatment did not affect the development of AHR. Immunization and aerosol challenges with OVA following the same protocol failed to induce AHR in the mast cell-deficient W/Wv mice, but induced AHR in their congenic littermates, i.e., WBB6F1 (+/+) mice. No significant difference was found between the W/Wv mice and +/+ mice with respect to the IgE and IgG1 anti-OVA Ab responses and to the airway eosinophilia after Ag provocations. It was also found that reconstitution of W/Wv mice with bone marrow-derived mast cells cultured from normal littermates restored the capacity of developing Ag-induced AHR, indicating that lack of mast cells was responsible for the failure of W/Wv mice to develop Ag-induced AHR under the experimental conditions. However, the OVA-immunized W/Wv mice developed AHR by increasing the frequency and Ag dose of bronchoprovocations. The results suggested that AHR could be developed by two distinct cellular mechanisms. One would go through mast cell activation and the other is IgE/mast cell independent but an eosinophil/IL-5-dependent mechanism.

Preoperative concurrent chemoradiotherapy against muscle-invasive bladder cancer: results of partial cystectomy in elderly or high-risk patients.

BACKGROUND: Good local control has been reported in cases of muscle-invasive bladder cancer treated by chemoradiotherapy and transurethral resection (TUR). However, definitive irradiation or extensive chemotherapy is often intolerable for elderly or poor-risk patients. We report here benefits of partial cystectomy after concurrent low-dose chemoradiotherapy for high-risk patients. METHODS: Thirty-seven patients with localized muscle-invasive bladder cancer (T2-T4) were treated with concurrent cisplatin (50-100 mg/body x 2 courses) and pelvic irradiation (40 Gy) preoperatively. Among 17 patients (46%) who achieved complete response (CR), 10 were not suitable for radical cystectomy and underwent partial cystectomy. Radical cystectomy was performed in 24 cases [CR = 6, partial response (PR) = 18]. Two patients (one CR and one PR) rejected open surgery and were treated by TUR of the primary site. One no change (NC) patient received no further treatment because of mental disorder. RESULTS: Median follow-up was 12 months (range 2-37 months). Fifteen of 36 evaluable cases (42%) achieved a pathological T0 response (no residual tumor). Estimated 3-year disease-free survival was 56% for all patients and 100% for T0 responders. Seven of 21 patients with pathological persistent tumor developed local recurrence (three patients) or distant metastasis (four patients). All of the 10 patients (eight with T0 response and two with a small residual tumor nest) who underwent partial cystectomy were recurrence-free for an observation period of up to 3 years. CONCLUSIONS: Bladder preservation by partial cystectomy may be a choice for patients who show a good response to preoperative chemoradiotherapy and are not suitable for radical cystectomy.

Production of endothelin by canine prostatic epithelial cells and its stimulatory effects on their growth.

The current study has been designed to explore expressions of endothelin (ET) receptors and ETs in the canine prostate and the effect of ETs on canine prostatic epithelial cells. ET receptors were characterized by biotinylated ET-1 binding in frozen sections of the prostate. Canine prostatic epithelial cells in primary culture were used for demonstration of ET-1 expression by reverse-phase HPLC coupled with radioimmunoassay and Northern blotting and were subjected to growth assay. Biotinylated ET-1 binding was localized in the epithelial component, and the binding was also blocked with an antagonist specific for ET(B) subtype receptors. ET-1 and ET-3 stimulated canine prostatic epithelial cell growth in vitro. The effect was also reversed in the presence of an antagonist specific for ET(B) subtype receptors. Elution profile of epithelial cell culture medium revealed two peaks, corresponding to ET-1 and big ET-1. Epithelial cells in culture expressed pre-pro-ET-1 mRNA. Canine prostatic epithelial cells expressed ET(B) receptors and ET-1. It appears most likely that the expressed ET-1 acts as an autocrine/paracrine proliferative factor on canine prostatic epithelial cells via ET(B) receptors.

Possible misinterpretation on computed tomography of left inferior vena cava as retroperitoneal lymph node metastasis: a report of two cases.

PURPOSE: We report on two cases of retroperitoneal lymph node metastasis of testicular cancer with left inferior vena cava. METHODS/RESULTS: A 25-year-old man with a left testicular cancer with pulmonary and retroperitoneal lymph node metastases received three courses of VIP (etoposide, ifosfamide and cisplatinum) chemotherapy. Subsequent abdominal computed tomography (CT) revealed round lesions enhanced with contrast agent on both sides of the aorta inside the degenerated lymphadenopathy. These lesions were regarded as a duplicated inferior vena cava (IVC) and this was confirmed at retroperitoneal lymph node dissection. The second case is of a 21-year-old man with a left testicular cancer with pulmonary, liver and widespread lymph node metastases. Subsequent to a course of VIP chemotherapy, super high-dose chemotherapy was administered. Abdominal CT revealed a round mass enhanced with contrast agent on the left side of the aorta adjacent to the degenerated lymphadenopathy, which was regarded as the transposed left IVC and this was confirmed at lymph node dissection. CONCLUSIONS: In both cases, initial CT failed to detect the lesions as the left IVC and there was a possibility for the misinterpretation of such venous anomalies with residual lymphadenopathy.

True hermaphroditism associated with microphthalmia.

A 4-year-old boy with an undescending left testis, penoscrotal hypospadia and bilateral microphthalmia was admitted to our hospital. Chromosome analysis revealed a karyotype of 46, XX del(x)(p2 2,31) and the sex-determining region of the Y chromosome (SRY) was negative. The right testis was located in the scrotum and a left cystic ovary-like gonad, a salpinx and a unicorn uterus were found in the left inguinal canal. Histologically the gonad was an ovotestis in which primordial follicles covered infantile seminiferous tubules. Microphthalmia is observed in some congenital syndromes caused by interstitial deletion of the X chromosome. This case suggested that the short arm of the X chromosome was involved in the differentiation of the gonad. Very closely located follicles and infantile seminiferous tubules indicated that induction of meiosis in the fetus was controlled by the local microenvironment in follicles and seminiferous tubules, and not by the systemic hormonal condition.

Immunosuppressive activities of recombinant glycosylation-inhibiting factor mutants.

We have shown previously that glycosylation-inhibiting factor (GIF) in culture supernatants of suppressor T cell (Ts) hybridomas had bioactivity, while the same cells contained a substantial quantity of inactive GIF in cytosol. Mass-spectrometric analysis of GIF in the culture supernatant and cytosol of a Ts hybridoma provided direct evidence that GIF protein was posttranslationally modified in the Ts cells, and that the GIF bioactivity is associated with the posttranslationally modified species. Assuming that conformational changes induced by the posttranslational modifications are responsible for generation of bioactivity, we constructed cysteine mutants of human rGIF (rhGIF) in which cysteine at position 57, 60, or 81 was replaced with Ala, and the mutants were expressed in Escherichia coli. Replacement of Cys57 or Cys60 with Ala resulted in generation of bioactivity, while replacement of Cys81 with Ala failed to do so. It was also found that replacement of Cys57 with Ala and carboxymethylation of a sulfhydryl group in Cys60 synergistically increased the GIF bioactivity of the GIF derivatives. A mutated GIF protein, in which Cys57 and Asn106 in the rhGIF were replaced with Ala and Ser, respectively, had immunosuppressive effects on the IgE and IgG1 Ab responses of BDF1 mice to DNP-OVA, while wild-type rhGIF did not. Evidence was obtained that the mutated GIF suppressed Ag priming of Th cells for the Ab responses and proliferative response.

Advanced testicular germ cell tumor in a hemophilic patient with human immunodeficiency virus infection.

A stage IIIB anaplastic seminoma which occurred in an HIV-infected hemophilia is reported. The patient with hemophilia A was 36 years old and had been seropositive for HIV antibody for 3 years. Inguinal orchiectomy and subsequent chemoradiotherapy for retroperitoneal lymphadenopathy were performed and a marker negative partial response was obtained. In spite of a low initial CD4+ lymphocyte count (90/microliter), the patient tolerated the treatment well without life-threatening opportunistic infection. Although factor VIII supplement was performed, continuous bleeding from the operative wound made postoperative care difficult.

Secondary treatment failure without anti-human chorionic gonadotropin antibody in a patient with Kallmann syndrome.

A 29-year-old man with Kallmann syndrome suddenly developed decreased semen volume, azoospermia, and facial hair loss after 11 years of successful human chorionic gonadotropin (hCG) and human menopausal gonadotropin (hMG) treatment. Anti-hCG antibody was not detected in the patient's serum. A high serum level of luteinizing hormone (LH) with nasal LH-releasing hormone analogue administration failed to increase serum testosterone to a sufficient level. Testosterone injection after cessation of hCG and hMG therapy was able to improve semen volume, but not azoospermia. Resumption of hCG and hMG therapy after 6 months cessation partially restored spermatogenesis. The secondary failure of hCG and hMG therapy suggests a decrease of testicular sensitivity to LH as well as hCG.

[High dose chemotherapy with peripheral blood stem cell transplantation (PBSCT) For advanced testicular cancer].

Five patients with metastatic testicular cancer of advanced extent according to the Indiana University criteria were enrolled into this study. All tumors were non-pretreated non-seminomas. Initially all patients were treated with standard dose etoposide, ifosfamide and cisplatin (VIP) regimen. The response of two cycles of VIP was evaluated by tumor markers and diagnostic imagings. Two of the five patients showed a good response to VIP and subsequently achieved a pathological complete response (pCR) following surgical resection of residual masses after 3 or 5 courses of VIP. However, they suffered from severe myelosuppression and underwent peripheral blood stem cell transplantation (PBSCT) following the final course of VIP. The remaining three patients unlikely to be cured by VIP underwent chemotherapy consisting of high dose ICE:ifosfamide (6-10 g/m2 over 4days) carboplatin (1,500 mg/m2 over 4 days), etoposide (1,600-2,400 mg/m2 over 4 days) combined with PBSCT. This regimen resulted in one partial response (PR) with marker-negative and two PR with marker-positive. Residual masses were removed in all three patients and viable tumor cells were found in two. Of the five patients enrolled, four patients (80%) remain disease-free with minimal follow-up of 20 months, and the remaining one died of cancer 10 months after PBSCT. No serious side effects or complications were encountered. This study shows that standard dose induction therapy of VIP followed by early salvage chemotherapy of high dose ICE with PBSCT is well tolerated and effective in the treatment of advanced poor-risk testicular cancer.

Significance of bladder neck involvement on progression in superficial bladder cancer.

OBJECTIVES: To determine the significance of bladder neck involvement in predicting disease progression in superficial (stage Ta and T1) transitional cell carcinoma (TCC) of the urinary bladder. PATIENTS AND METHODS: A series of 277 patients with newly diagnosed superficial TCCs of the bladder was reviewed, and disease progression (to T2 or worse) was considered. The significance of several risk factors including bladder neck involvement was assessed in univariate and multivariate analysis. RESULTS: Progression occurred in 28 (10.1%) of 277 patients during a median follow-up period of 7.7 years. Nineteen died of bladder cancer. The following variables were found to be statistically significant at the univariate analysis (p < 0.05): irritative symptoms, urine cytology, tumor stage, involvement of the bladder neck, and tumor grade. Indeed, only involvement of the bladder neck, tumor stage, and grade retained their value as independent factors for progression at multivariate analysis. Patients were divided into three groups according to the number of independent risk factors they had. Groups having none, one, and two or three risk factors included 129, 99, and 49 patients with 5-year progression rates of 0.8, 4.6 and 27.5%, and 15-year rates of 4.0, 20.1 and 42.7%, respectively. CONCLUSION: Involvement of the bladder neck is a significant and independent risk factor for progression of superficial TCCs in addition to the histologic grade and stage. The combination of these three risk factors offers better prediction of progression in an individual patient.