OBJECTIVE: The objective of this study was to investigate the significance of the activated Akt-mammalian target of rapamycin (Akt-mTOR) signaling pathway in the progression of prostate cancer. MATERIALS AND METHODS: The expression levels of Akt, phosphorylated Akt (p-Akt), mTOR and phosphorylated mTOR (p-mTOR) in 175 prostate specimens, including 61 normal prostate tissues as a control, 24 high-grade prostatic intraepithelial neoplasias (HGPINs) and 90 clinically localized prostate cancers, were evaluated by immunohistochemical staining. p-Akt and p-mTOR ratios, which were defined as the expression level of p-Akt in relation to that of Akt and the expression level of p-mTOR in relation to that of mTOR, respectively, in these specimens were calculated. RESULTS: Expression levels of all four molecules, including Akt, p-Akt, mTOR and p-mTOR, were significantly greater in the HGPIN group compared with the normal control and prostate cancer groups. Furthermore, the p-Akt ratio in the prostate cancer group was significantly lower than that in the HGPIN group, while there was no significant difference in the p-mTOR ratio between the HGPIN and prostate cancer groups. In the prostate cancer group, no significant relationships were observed between major clinicopathological parameters and the expression levels as well as the ratios of p-Akt or p-mTOR. CONCLUSIONS: The Akt-mTOR signaling pathway may play a limited role in the progression of prostate cancer

OBJECTIVE: Sonic hedgehog (Shh) signaling, androgens and epithelial-mesenchymal transition (EMT) are related to prostate cancer (PCa) progression. The aim of this study was to investigate how Shh and androgen [dihydrotestosterone (DHT)] signaling act in prostate epithelial and stromal compartments and whether this signaling pathway drives EMT and promotes PCa progression. MATERIAL AND METHODS: LNCaP, normal prostate fibroblast (NPF) and cancer-associated prostate fibroblast (CPF) cells were studied with DHT and/or the Shh signaling inhibitor cyclopamine. Real-time reverse transcription-polymerase chain reaction (RT-PCR) was performed to evaluate the expressions of a potential Shh target gene, osteonectin (ON) and EMT-associated markers (E-cadherin, N-cadherin and vimentin). Immunohistochemical studies using PCa prostatectomy samples were performed to assess the expression levels of ON, Gli-1, androgen receptor, Shh, E-cadherin, N-cadherin and vimentin. RESULTS: While DHT enhanced cell proliferation in CPF more than LNCaP or NPF, cyclopamine inhibited cell proliferation enhanced by DHT in CPF. Real-time RT-PCR showed whereas both Shh and DHT induced N-cadherin and vimentin, DHT also induced the expression of osteonectin in LNCaP and cyclopamine blocked these expressions in osteonectin, N-cadherin and vimentin (p = 0.0084, 0.0002 and 0.0373, respectively). Immunohistochemistry showed that high expression of stromal, but, not epithelial, ON was significantly correlated with serum prostate-specific antigen (PSA) (p = 0.031), and high expression of Gli-1 and low expression of stromal ON with PSA recurrence (p = 0.0114 and p = 0.0005, respectively). CONCLUSIONS: Shh and androgen signaling in prostate tumor and stromal compartments drives EMT, and thus may play some role in PCa progression. Cyclopamine may be one therapeutic strategy for PCa.

OBJECTIVE: To compare the quality of surgical management of the neurovascular bundle (NVB) by assessment of neuronal nitric oxide synthase (nNOS)-positive nerves in surgical specimens between open retropubic radical prostatectomy (RP; ORRP) and robot-assisted RP (RARP). METHODS: This study included 65 (99 sides, NVB resection; 31 sides, NVB preservation) and 83 (106 sides, NVB resection; 60 sides, NVB preservation) patients undergoing ORRP and RARP, respectively. The posterior sectors from the apex, mid, and base areas on each side were immunohistochemically stained with an nNOS antibody. RESULTS: On the sides with NVB resection, there were no significant differences in the numbers of nNOS-positive nerves in any areas between the ORRP and RARP groups; however, on the sides with NVB preservation, the numbers of nNOS-positive nerves in the ORRP group were significantly higher than those in the RARP group at the apex (84.4 vs 59.2; P = .0028), mid (71.2 vs 52.4; P = .016), and base (148.0 vs 40.8; P .001) areas. In 55 patients who were judged not to have severe erectile dysfunction before surgery and subsequently underwent nerve-sparing RP, there was a significantly inverse correlation between the total number of nNOS-positive nerves on both sides and the postoperative erectile function. CONCLUSION: These findings suggest that RARP might be suitable for performing precise nerve-sparing surgery compared with ORPP, particularly in the base area of the prostate and that the quantification of nNOS-positive nerves in surgical specimens could be a useful approach for predicting the postoperative erectile function.

BACKGROUND: To evaluate the expression of multiple molecular markers involved in the mammalian target of rapamycin (mTOR) signaling pathway in human muscle-invasive bladder cancer (BC) and to assess the therapeutic efficacies of mTOR inhibitors in human BC KoTCC-1 cells. METHODS: Expression levels of 5 markers, including PTEN, phosphorylated (p)-Akt, p-mTOR, p-p70 ribosomal S6 kinase, and p-4E-binding protein 1 (4E-BP1), were measured in radical cystectomy specimens from 49 patients with muscle-invasive BC by immunohistochemical staining. We then analyzed the effects of treatment with temsirolimus or Ku-0063794, a dual inhibitor of mTOR complex 1 (C1) and mTOR complex 2 (C2), on changes in the growth and expression profiles of 5 mTOR-associated markers in KoTCC-1 cells. RESULTS: During the follow-up period of this study, disease recurred in 27 patients (55.1%), and of several factors examined, the expression level of p-4E-BP1 in addition to the pathological T stage was independently related to recurrence-free survival on multivariate analysis. Although the growth of KoTCC-1 cells was inhibited by both temsirolimus and Ku-0063794 in dose-dependent manners, treatment with Ku-0063794 resulted in a marked decrease in the expression of p-4E-BP1 in KoTCC-1 cells compared with that with temsirolimus. Furthermore, the growth-inhibitory effect of both mTOR inhibitors was shown to be proportional to the expression levels of p-4E-BP1. CONCLUSIONS: The phosphorylation status of 4E-BP1 appeared to be correlated with the prognosis of patients with muscle-invasive BC following radical cystectomy as well as the sensitivities of BC cells to mTOR inhibitors; therefore, the inactivation of 4E-BP1 using Ku-0063794 may be a promising novel approach for muscle-invasive BC.

The treatment failure often happens in overactive bladder (OAB) partly owing to its unknown pathogenesis. The purpose of this study is to find significant receptors or biological markers for OAB-related symptoms for establishment of potential order-made therapeutic strategies. The overactive bladder symptom scores (OABSS) and international prostate symptom scores (IPSS)/quality of life (QOL) were questioned in all the 18 patients with OAB diagnosis. Their bladder mucosal tissues were taken from the random biopsy of bladder cancer suspected patients without any finding such as inflammation or carcinoma in situ. They were investigated quantitatively by immunohistochemical (IHC) stainings for inflammatory or immune-system (Interleukin (IL)-6 and cyclooxygenase-2 (Cox-2)), Caspase-3 apoptosis markers, angiogenesis (CD-31), epithelial-mesenchymal transition (E-cadherin) and muscarinic receptor (Muscarine-2 (M)-2), adrenergic receptors (ARs) (alpha 1-d (α1-d) and beta-3 (β-3)). The statistical correlation between the expressions of these 5 markers and 3 receptors and these symptom scores were examined under the comparison between OAB patients and control patients who had urgency score with less than 2 in OABSS. The OABSS and IPSS/QOL was 7.39 ± 2.69 and 21.2 ± 6.59/4.33 ± 1.33, respectively but those of control patients were 2.00 ± 1.41 and 10.1 ± 9.52/2.14 ± 1.46, respectively (P0.05). Regarding the correlation of those markers' expressions and symptom scores, in OAB patients, OABSS total significantly correlated with β-3 AR expressions (P=0.0457). IPSS post-voiding significantly correlated with β-3 AR expressions (P=0.0308) but no significant relationship in control patients (P>0.05). In conclusion, this study demonstrated that β-3 AR in our tested 8 markers or receptors was correlated strongly with OAB-related symptoms. These data may help elucidate the pathophysiology of OAB and offer possible strategy for its order-made therapies.

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Oocysts of C. muris and the events of excystation using 0.5% sodium hypochlorite as excystation medium were described with light microscope. The response of the immunocompetent BALB/c mice against infection was studied using sera of orally infected mice at different periods postinoculation by indirect immunofluorescence antibody test using 1:150 FITC conjugated rabbit serum antimouse polyclonal IgG. From the patterns of IFAT, it was suggested that the dominant antigen in C. muris was restricted to the apical complex of the sporozoites. Such antigen may play a role in the invasion of the host cell. Future analysis of such receptor molecules might constitute prime candidates as immunogens for a vaccine, the efficiency of which might cause inhibition of parasite invasion.

In the present study, wes adapted for detection of infection in snail by ELISA TEST , previously shown by us to be very sensitive for detecting cercariae in water, for the sensitive detection of Schistosoma mansoni abs in infected snails from early prepatency. Polymerase chain reaction primers were designed based on the

Abstract A double antibody sandwich ELISA technique, using a chromatography purified antisera against E. histolytica, G. lamblia and Cryptosporidium antigens, was applied to detect copro-antigens of the corrosponding parasites in 90 patients. All positive cases were diagnosed by parasitological examination and proved to have the infection solely. Beside the 90 positive cases, 40 age-matched controls were included in the study, of which 20 individuals were infected with other parasites but not Cryptosporidium, E. histolytica or G. lamblia (acted as an infected control group) and the other 20 individuals with no intestinal parasites (normal control group). The assay could detect 100% of those infected with both of G. lamblia and E. histolytica and 96.6% (29/30) of patients with Cryptosporidium infection. False positive reactions were detected in 3 cases using G. lamblia antisera (92.5%), 5 cases using E. histolytica antisera (87.5%) and 2 cases using Cryptosporidium antisera (95%). A direct increase in the mean antigen level was observed with the increasing intensity of infection in the 3 parasites, so higher mean O.D. readings was observed in heavily infected cases than moderately infected cases than lighter intensity of infection. Only those in elder age group (> 20 years) infected with E. histolytica were found to have statistically higher O.D. readings of the antigen than middle age group (10-20 years). On the other hand, no statistically significant difference was observed between different age groups and antigen level in cases with either G. lamblia or Cryptosporidium.