HGF-induced capillary morphogenesis of endothelial cells is regulated by Src.

The signal transduction pathway involved in hepatocyte growth factor (HGF)-induced capillary morphogenesis of endothelial cells was investigated. HGF-induced capillary morphogenesis of the murine spleen endothelial cell line MSS31 was inhibited by a Src family kinase inhibitor, PP2. Stable expression of kinase-inactive Src in MSS31 cells inhibited HGF-induced activation of Src as well as capillary morphogenesis. The HGF-induced capillary morphogenesis of human umbilical vein endothelial cells was also inhibited by PP2 and was reduced by the downregulation of Src by small interfering RNA. These results suggest that HGF induces capillary morphogenesis of endothelial cells through Src

Cryoimmunologic Investigations in Patients With Prostatic Carcinoma

Transurethral cryosurgery was performed with liquid nitrogen in 18 patients with prostatic carcinomas. The clinical course and the effect of cryosurgery on immune competence of patients were observed. After cryosurgery, pain was relieved in 5 of 6 patients, dysuria was improved in 7 of 12 patients, and the prostatic mass was reduced in 14 of 18 patients. Elevated serum acid phosphatase levels returned to normal in 5 cases. Changes of serum protein fractions were variable. Serum complement levels were high in advanced cases. Changes of immunoglobulins were variable. Serum anti-prostatic antibody could not be detected by passive hemagglutination test and by indirect immunofluorescent examination. DNCB skin reaction and blastoid transformation of lymphocytes were raised after therapy. The cellular immune competence may be enhanced by cryosurgery, while the humoral immune competence is scarcely influenced by the therapy. It has been reported that remission of metastatic lesions occured after cryosurgery for advanced prostatic carcinoma (Soanes et al.1)). Furthermore, it was suggested that immune competence of the patient with carcinoma may be enhanced after cryosurgery (Ablin2)). In an attempt to examine the effect of cryosurgery on immune competence, we observed clinical and immunologic responses after treatment in 18 patients with prostatic carcinoma

Elevated Expression of Pyrimidine Nucleoside Phosphorylase (PyNPase) in Renal Cell Carcinoma Tissue

Background: Pyrimidine nucleoside phosphorylase (PyNPase) is an enzyme that converts 5\u27-deoxy-5-fluorouridine to 5- fluorouracil. PyNPase is identical to platelet-derived endothelial cell growth factor (PD-ECGF), and has angiogenic activity. In the present study of PyNPase activity in renal cell carcinoma, we tested for correlation between PyNPase activity and tumor extension, tumor proliferation and clinical characteristics. Methods: Samples of tumor tissue and non-tumor tissue were obtained from 10 renal cell carcinoma patients who underwent radical nephrectomy. These samples were examined, and PyNPase activity of the tissues was assayed. Results: PyNPase activity was significantly higher in renal cancer tissue than in normal tissue (p<0.01), and in immunohistochemical assays PyNPase was strongly stained in the cytoplasm of renal cancer cells. However, there were no significant correlation between PyNPase activity and tumor extension, according to the results of histopathological examination and evaluation of vascularity of renal cancer tumor tissue. Conclusion: In this study, we did not observe correlation between PyNPase activity and renal cancer extension and proliferation. However, the present data suggest that pyrimidine-class drugs may be useful against renal cell carcinoma, because PyNPase activity is significantly higher in renal cancer tissue than in normal tissue

Upregulation of CA 19-9 in the Mouse Kidney Following Unilateral Ureteral Obstruction

High serum levels of carbohydrate antigen19-9 (CA19-9) have been detected in patients with pancreatic cancer and described in several recent case reports of patients with hydronephrosis. However, the mechanism of high serum levels of CA19-9 among hydronephrosis cases remains to be elucidated. In this study, we established a mouse unilateral ureteral obstruction (UUO) model to investigate the expression of CA19-9 protein in renal tissue. To investigate the progression of hydronephrosis following UUO, MR urography and pathological analysis were performed. CA19-9 expression was examined by immunohistochemistry and western blot analysis. MR urography revealed that the grade of pelvic dilatation increased in a time dependent manner. Pathologically, both interstitial cellular infiltration and fibrosis were detected from the second to the fourteenth day after surgery in UUO mice. CA19-9 was detected in the UUO kidney after the second day. The immunoblot analysis revealed that the elevated expression of CA19-9 was demonstrated at an early stage of obstructive nephropathy. Our study shows that the ureteral obstructed kidney is dominated by cell infiltration and induced fibrosis. The selective expression of CA19-9 was detected in renal fibrous tissue. Based on these findings, the level of CA19-9 might be a good indicator for onset of renal fibrosis induced by obstruction

Immunohistochemical Analysis of Connexin43 Expression in Infertile Human Testes

Connexin43 (Cx43) is abundantly expressed in mammalian testes and implicated in the regulation of cell-to-cell interaction between germ cells and Sertoli cells, which is essential to the normal process of spermatogenesis. In the present study, we investigated the relation between Cx43 expression and the degree of spermatogenesis in infertile human testes. Immunohistochemical analysis of Cx43 was performed on testicular biopsies from 29 patients with azoospermia (n=23) and severe oligospermia (n=6), who gave informed consent to this experiment. The degree of testicular spermatogenesis was evaluated by Johnsen score. In the interstitium, immunostaining for Cx43 was localized to some focal parts of plasma membrane between neighboring Leydig cells. In seminiferous tubules with normal spermatogenesis, Cx43 expression was found between Sertoli cells and germ cells. However, Cx43 expression in maturation arrest was decreased and located mainly in the basal compartment of seminiferous tubules. Finally, there was a significant positive correlation between histological score of spermatogenesis and intensity of Cx43 (p=0.0294). These data suggest that the alteration of Cx43 expression may be involved in spermatogenic impairment, and that the communication between Sertoli cells and germ cells through Cx43 may be important for maturation of spermatogenesis

Small Cell Carcinoma of the Ureter with Malignant Lymphoma: Case Report and Literature Review

A 78-year-old man was referred to our hospital for gross hematuria. Ultrasonography and magnetic resonance urography revealed tumor in the right lower ureter. Computed tomography revealed right cervical lymph node swelling and pathological diagnosis was diffuse large B-cell lymphoma. Right nephroureterectomy was performed and pathologic examination revealed small cell carcinoma of the ureter with a small urothelial cell carcinoma component. As the patient had concomitant other malignancies, additional systemic chemotherapy was not performed. As of 3 months after operation, postoperative course has been uneventful. Only 11 cases of primary small cell carcinoma of the ureter have been described

Downregulation of Fes inhibits VEGF-A-induced chemotaxis and capillary-like morphogenesis by cultured endothelial cells.

The aim of this study was to determine whether the downregulation of endogenous Fes by siRNA in cultured endothelial cells affects vascular endothelial growth factor-A (VEGF-A)-induced chemotaxis and capillary-like morphogenesis, which are considered as angiogenic cellular responses in vitro. VEGF-A-treatment induced autophosphorylation of Fes in cultured endothelial cells. LY294002, a phosphoinositide 3-kinase inhibitor, significantly inhibited VEGF-A-induced chemotaxis and capillary-like morphogenesis. Downregulation of Fes attenuated these VEGF-A-induced cellular responses but LY294002 did not produce further inhibition of these responses. Downregulation of Fes neither affected VEGF-A-induced autophosphorylation of VEGF receptor 2 nor mitogen-activated protein kinase activation, but markedly decreased Akt activation. Taken together, our novel results indicate the involvement of Fes in VEGF-A-induced cellular responses by cultured endothelial cells

Elevated expression of ERK 2 in human tumor cells chronically treated with PD98059.

We examined the effect of chronic exposure of tumor cells to a mitogen-activated protein kinase/extracellular signal-regulated kinases (ERK) kinase inhibitor, PD98059, on cell proliferation was investigated. Human renal carcinoma cells (ACHN) and prostatic carcinoma cells (DU145) were cultured in the presence of PD98059 for more than 4 weeks (denoted ACHN (PD) cells and DU145 (PD) cells, respectively) and proliferation and signal transduction pathways were examined. PD98059 significantly inhibited the proliferation of parental cells. However, PD98059 failed to inhibit proliferation of ACHN (PD) and DU145 (PD) cells significantly. Expression of ERK 1 and 2 was elevated in these cells. These phenotypes were reversible. Downregulation of ERK 2, but not ERK 1, by small interfering RNA significantly inhibited the proliferation of ACHN (PD) and DU145 (PD) cells. Taken together, chronic exposure of tumor cells to PD98059 induced elevated expression of ERK 2, which was associated with decreased sensitivity of cellular proliferation to PD98059

Semen quality of 1559 young men from four cities in Japan: a cross-sectional population-based study

Objectives: To provide information of semen quality among normal young Japanese men and indicate the frequency of reduced semen quality. Design: Cross-sectional, coordinated studies of Japanese young men included from university areas. The men had to be 18-24 years, and both the man and his mother had to be born in Japan. Background information was obtained from questionnaires. Standardised and quality-controlled semen analyses were performed, reproductive hormones analysed centrally and results adjusted for confounding factors. Setting: Four study centres in Japan (Kawasaki, Osaka, Kanazawa and Nagasaki). Participants: 1559 men, median age 21.1 years, included during 1999-2003. Outcome measures: Semen volume, sperm concentration, total sperm count, sperm motility, sperm morphology and reproductive hormone levels. Results: Median sperm concentration was 59 (95% CI 52 to 68) million/ml, and 9% and 31.9% had less than 15 and 40 million/ml, respectively. Median percentage of morphologically normal spermatozoa was 9.6 (8.8 to 10.3)%. Small, but statistically significant, differences were detected for both semen and reproductive hormone variables between men from the four cities. Overall, the semen values were lower than those of a reference population of 792 fertile Japanese men. Conclusions: Assuming that the investigated men were representative for young Japanese men, a significant proportion of the population had suboptimal semen quality with reduced fertility potential, and as a group they had lower semen quality than fertile men. However, the definitive role-if any-of low semen quality for subfertility and low fertility rates remain to be investigated

Fibroblast growth factor-2 induces the activation of Src through Fes, which regulates focal adhesion disassembly.

Cell migration is regulated by focal adhesion (FA) turnover. Fibroblast growth factor-2 (FGF-2) induces FA disassembly in the murine brain capillary endothelial cell line IBE, leading to FGF-2-directed chemotaxis. We previously showed that activation of Src and Fes by FGF-2 was involved in chemotaxis of IBE cells. In this study, we examined the interplay between Src and Fes. FGF-2 treatment decreased the number of FA in IBE cells, but not in cells expressing dominant-negative Fes (denoted KE5-15 cells). FGF-2 induced the activation of Src and subsequent binding to and phosphorylation of Cas in IBE cells, but not in KE5-15 cells. Focal adhesion kinase (FAK) activation and tyrosine phosphorylation by Src were also delayed in KE5-15 cells compared to parental cells. FGF-2 induced activation of Src within FA in IBE cells, but not in KE5-15 cells. Downregulation of Fes or FAK using small interfering RNA diminished Src activation by FGF-2 within FA. These findings suggest that activation of Fes by FGF-2 enhances FAK-dependent activation of Src within FA, promoting FGF-2-induced disassembly of focal adhesions