Soy Isoflavones Induce Feminization of Japanese Eel (Anguilla japonica).

Under aquaculture conditions, Japanese eels (Anguilla japonica) produce a high percentage of males. However, females gain higher body weight and have better commercial value than males, and, therefore, a high female ratio is required in eel aquaculture. In this study, we examined the effects of isoflavones, genistein, and daidzein on sex differentiation and sex-specific genes of eels. To investigate the effects of these phytoestrogens on the gonadal sex, we explored the feminizing effects of soy isoflavones, genistein, and daidzein in a dose-dependent manner. The results showed that genistein induced feminization more efficiently than daidzein. To identify the molecular mechanisms of sex-specific genes, we performed a comprehensive expression analysis by quantitative real-time PCR and RNA sequencing. Phenotypic males and females were produced by feeding elvers a normal diet or an estradiol-17ß- or genistein-treated diet for 45 days. The results showed that female-specific genes were up-regulated and male-specific genes were down-regulated in the gonads, suggesting that genistein induces feminization by altering the molecular pathways responsible for eel sex differentiation.

Profiles of 5α-Reduced Androgens in Humans and Eels: 5α-Dihydrotestosterone and 11-Ketodihydrotestosterone Are Active Androgens Produced in Eel Gonads.

Although 11-ketotestosterone (11KT) and testosterone (T) are major androgens in both teleosts and humans, their 5α-reduced derivatives produced by steroid 5α-reductase (SRD5A/srd5a), i.e., 11-ketodihydrotestosterone (11KDHT) and 5α-dihydrotestosterone (DHT), remains poorly characterized, especially in teleosts. In this study, we compared the presence and production of DHT and 11KDHT in Japanese eels and humans. Plasma 11KT concentrations were similar in both male and female eels, whereas T levels were much higher in females. In accordance with the levels of their precursors, 11KDHT levels did not show sexual dimorphism, whereas DHT levels were much higher in females. It is noteworthy that plasma DHT levels in female eels were higher than those in men. In addition, plasma 11KDHT was undetectable in both sexes in humans, despite the presence of 11KT. Three srd5a genes (srd5a1, srd5a2a and srd5a2b) were cloned from eel gonads. All three srd5a genes were expressed in the ovary, whereas only both srd5a2 genes were expressed in the testis. Human SRD5A1 was expressed in testis, ovary and adrenal, whereas SRD5A2 was expressed only in testis. Human SRD5A1, SRD5A2 and both eel srd5a2 isoforms catalyzed the conversion of T and 11KT into DHT and 11KDHT, respectively, whereas only eel srd5a1 converted T into DHT. DHT and 11KDHT activated eel androgen receptor (ar)α-mediated transactivation as similar fashion to T and 11KT. In contrast, human AR and eel arß were activated by DHT and11KDHT more strongly than T and 11KT. These results indicate that in teleosts, DHT and 11KDHT may be important 5α-reduced androgens produced in the gonads. In contrast, DHT is the only major 5α-reduced androgens in healthy humans.

Tumor Location and Pathological Features of Latent and Incidental Prostate Cancer in Contemporary Japanese Men.

PURPOSE: The incidence of prostate cancer is increasing in Asian countries. Studies using prostatectomy specimens have reported a racial difference in tumor location within the prostate, with a greater incidence of transition zone cancer in Asian men compared with Caucasian men. However, there may be potential biases in studies based on surgical specimens. We describe the pathological features of subclinical prostate cancer, such as latent cancer and incidental cancer, to elucidate tumor location of contemporary Japanese patients. We also compare the prevalence of latent and incidental prostate cancer to determine whether the incidence of prostate cancer is higher in patients with bladder cancer. MATERIALS AND METHODS: Overall 182 men autopsied and 148 who underwent cystoprostatectomy for bladder cancer were included in the study. Each prostate gland was fixed and sliced in step sections. Histological evaluation was performed by a single genitourinary pathologist. The index tumor location was categorized into transition zone or peripheral zone. RESULTS: Prostate cancer was found in 39.0% of the autopsy specimens and 31.6% of the cystoprostatectomy specimens. The prevalence and pathological characteristics were not significantly different between latent and incidental cancer. The prevalence of transition zone cancer was 39.0% (46 of 118). In elderly men peripheral zone cancer was more frequently diagnosed than transition zone cancer (p=0.049). The pathological characteristics of transition and peripheral zone cancers were similar except for the pT stage. CONCLUSIONS: Transition zone cancer was prevalent in contemporary Japanese men. The incidence of prostate cancer in men with bladder cancer might not be higher than that in healthy men.

Red Cell Distribution Width Predicts Prostate-Specific Antigen Response and Survival of Patients With Castration-Resistant Prostate Cancer Treated With Androgen Receptor Axis-Targeted Agents.

PURPOSE: To identify the impact of red cell distribution width (RDW) on treatment outcomes in patients with castration-resistant prostate cancer (CRPC) treated with androgen receptor axis-targeted agents (ARATs). PATIENTS AND METHODS: Baseline data were obtained from 153 patients with CRPC treated with ARATs. Patients were stratified according to the upper limit of the normal RDW range, measured within 1 month before starting treatment. Relationships between RDW levels and the best prostate-specific antigen (PSA) response, PSA progression-free survival, and overall survival were examined. RESULTS: Forty-nine patients were treated with abiraterone acetate in combination with corticosteroid and 104 with enzalutamide. The median RDW was 13.7% (interquartile range, 13.0-14.9). High RDW was significantly associated with prior use of docetaxel (P < .001), presence of lymph node metastasis (P = .031), presence of visceral metastasis (P = .001), and low hemoglobin (P < .001), low albumin (P = .016), and high C-reactive protein levels (P = .02). In a multiple linear regression model, there was a statistically significant negative association between RDW levels and the best PSA response (P = .046). In addition, multivariate Cox regression analyses showed that high RDW was an independent predictor of both shorter PSA progression-free survival (hazard ratio = 1.84; 95% confidence interval, 1.04-3.27; P = .037) and overall survival (hazard ratio = 2.62; 95% confidence interval, 1.15-5.98; P = .022), showing statistical significance. CONCLUSION: High RDW is an independent predictor of worse treatment outcomes in patients with CRPC treated with ARATs. RDW could be a readily available and inexpensive biomarker for predicting primary resistance to ARATs.

Hyperprogression after pembrolizumab treatment in two patients with metastatic urothelial carcinoma.

Hyperprogression has recently been recognized as a new pattern of progression in patients undergoing immune checkpoint inhibitor treatment. Here, we report two cases that showed hyperprogression during the initial phase of pembrolizumab treatment for metastatic urothelial carcinoma. The first patient, who received pembrolizumab as a second-line treatment, developed severe respiratory failure due to the rapid progression of lung metastases on the ninth day after the third pembrolizumab treatment. The second patient developed jaundice and hepatic dysfunction due to the progression of a metastatic lymph node of the liver hilum after the first administration of pembrolizumab. She developed multiple brain metastases with intraventricular bleeding on the 10th day after the second administration of pembrolizumab. It is important to be aware that hyperprogression sometimes occurs quite a while after starting treatment, and that both pseudoprogression and hyperprogression may occur in the early stage of treatment.

Comparison of ERG and SPINK1 expression among incidental and metastatic prostate cancer in Japanese men.

BACKGROUND: TMPRSS2:ERG fusion is the most common genetic event in prostate cancer (PCa). However, its association with prognosis is controversial. Overexpression of serine protease inhibitor Kazal-type 1 (SPINK1) was almost exclusively defined in ERG-negative PCa in most studies. This study aimed to determine the association between ERG and SPINK1 expression and the biological aggressiveness of PCa by analyzing their expression in incidental and metastatic cohorts. METHODS: A total of 143 cystoprostatectomy specimens of invasive bladder cancer and 98 biopsy specimens from de novo metastatic PCa were analyzed. The prostate gland of cystoprostatectomy specimens was fixed and sliced in step sections. Immunohistochemistry of ERG and SPINK1 was conducted, and the results were correlated with the clinicopathological characteristics of the patients. RESULTS: The overall prevalence of incidental cancer was 32.2% (46/143). The frequencies of both ERG and SPINK1 expression were not significantly different between incidental and metastatic cohorts (15.2% and 14.3%; P = 1.00, and 6.5% and 12.2%; P = 0.38, respectively). In the metastatic cohort, any pre-treatment factors were not significantly associated with the frequencies of ERG and SPINK1 expression. However, SPINK1 expression was significantly associated with a shorter time to castration-resistant PCa (CRPC) (P = 0.048). Meanwhile, overall survival was not significantly associated with the expression status of ERG and SPINK1 (P = 0.71). CONCLUSIONS: ERG and SPINK1 expression may not have significant influence on the metastatic behavior of PCa. SPINK1 expression was significantly associated with a shorter time to CRPC in metastatic PCa. The expression profile of ERG and SPINK1 may be a useful predictor for effect of androgen deprivation therapy in patients with metastatic castration-sensitive PCa.

Effect of everolimus treatment for regrown renal angiomyolipoma associated with tuberous sclerosis complex after transcatheter arterial embolization.

BACKGROUND: The aim of this study was to evaluate the effects and the utility of second-line everolimus treatment for regrown renal angiomyolipoma (AML) with tuberous sclerosis complex (TSC) after transcatheter arterial embolization (TAE). METHODS: We investigated a total of 14 patients who underwent second-line everolimus treatment for TSC-AML that regrew after TAE, and assessed their effects and adverse events. Everolimus treatment was performed for AML with a maximum diameter of 4 cm. To determine the reduction ratio of AML, the volume of AML was measured using multislice helical computed tomography. Adverse events were evaluated according to CTCAE v4.0-JCOG. We further compared the treatment effect and adverse events with those in patients receiving first-line everolimus treatment. RESULTS: The AML volume decreased in all patients, with a ≥ 50% volume decrease in 57% (8 of 14) of the cases, and the mean reduction rate was 53%. We observed no significant difference in the mean reduction rate of AML between second-line everolimus treatment for regrown TSC-AML after TAE and first-line everolimus treatment for TSC-AML. The adverse events were mild and consistent with those reported in our previous study. CONCLUSION: Although further studies are needed, everolimus appears to be effective as second-line treatment for TSC-AML that regrew after TAE and a beneficial treatment option for TSC-AML.

Effect of everolimus treatment for renal angiomyolipoma associated with tuberous sclerosis complex: an evaluation based on tumor density.

BACKGROUND: The aim of this study was to evaluate the influence of components of angiomyolipoma (AML) on the efficacy of everolimus. METHODS: We investigated a total of 40 patients with tuberous sclerosis complex (TSC) who had AML ≥4 cm in diameter. The components of the AML were determined using abdominal computed tomography (CT) images. The AML density was measured as the mean Hounsfield unit (HU) values of the whole area of the AML on axial CT images. We classified them into two groups, i.e., a lipid group with a predominant lipid component (HU ≤ -50) and a solid group with predominant vascular and muscle components (HU ≥30). For each patient, we measured the AML reduction rate and transition of the mean HU value. RESULTS: The mean reduction rate of AML in the lipid group was 24%, whereas it was 68% in the solid group (P < 0.001). The mean tumor density after 6 months was decreased in both groups. In particular, the density significantly decreased compared to the baseline in the solid group (P < 0.001). The tumor density did not change after 6 months in either group. CONCLUSION: The effect of everolimus on TSC-AML is mainly a reduction of the solid components consisting of angioma and leiomyoma. The tumor density at the start of treatment might be a predictive marker for the response to everolimus in TSC-AML.

Intermittent everolimus administration for renal angiomyolipoma associated with tuberous sclerosis complex.

OBJECTIVES: To evaluate the effects and utility of intermittent everolimus treatment for renal angiomyolipoma associated with tuberous sclerosis complex. METHODS: We investigated a total of 26 patients with tuberous sclerosis complex who had angiomyolipoma ≥4 cm in diameter. For each patient, we analyzed the reduction in the size of the angiomyolipoma, the change in size after everolimus withdrawal, the size reduction rate on everolimus readministration and adverse events caused by everolimus. The volume of angiomyolipoma was measured using abdominal computed tomography or magnetic resonance imaging. Adverse events were evaluated according to CTCAE v4.0-JCOG. RESULTS: The average size reduction rate of angiomyolipoma in the initial treatment with everolimus was 67%. Eight patients (31%) did not have enlarged angiomyolipoma after everolimus withdrawal. The other 18 patients (69%) restarted everolimus treatment because of enlargement of the angiomyolipoma. The average size reduction rate of angiomyolipoma in the everolimus retreatment group was 61%, which was equivalent to the rate for the initial treatment. There were fewer adverse events during everolimus retreatment than in the initial treatment. CONCLUSIONS: This is the first report regarding intermittent everolimus treatment for renal angiomyolipoma associated with tuberous sclerosis complex. This treatment is effective for tumor control and adverse event management. This beneficial treatment option for patients can minimize the drug dosage and the occurrence of adverse events.

Outcomes of everolimus treatment for renal angiomyolipoma associated with tuberous sclerosis complex: A single institution experience in Japan.

OBJECTIVES: To evaluate the efficacy and toxicity profiles of everolimus in Japanese patients with renal angiomyolipoma associated with tuberous sclerosis complex. METHODS: Patients with a 4-cm or larger angiomyolipoma meeting the diagnostic criteria of tuberous sclerosis complex were selected as participants of our investigation. In each case, we assessed tuberous sclerosis complex-associated symptoms, the treatment effect and adverse events. The treatment effect was evaluated by measuring the tumor volume reduction rate using abdominal computed tomography or magneitc resonance imaging. Adverse events were investigated using CTCAE v4.0-JCOG. The dose of everolimus was set at 10 mg once a day for adults. For childhood angiomyolipoma, everolimus administration was initiated at a dose of 5 mg once a day. Blood everolimus level was measured, and the dose was adjusted to maintain this within a range of 5-15 ng/mL. RESULTS: The angiomyolipoma volume decreased in 46 of 47 cases, and the mean reduction rate for all cases was 60% in 12 months. The angiomyolipoma volume markedly decreased in the first 3 months, and the size leveled off after 6 months. The main adverse events related to everolimus treatment were stomatitis (91%) and irregular menstruation (65%). Grade 3 or severer adverse events were noted in three cases (6%). All patients developed some adverse events in the first 6 months. The incidence markedly decreased to 40-50% after 13 months. CONCLUSION: The tumor volume-reducing effect of everolimus in a Japanese population was equivalent to or higher than that in Western populations. A wide variety of everolimus treatment-related adverse events can be observed, but most cases are very mild. Special attention should be given to stomatitis, irregular menstruation and interstitial lung disease as adverse events.

Time Trends in Histological Features of Latent Prostate Cancer in Japan.

PURPOSE: The incidence of prostate cancer is reported to be increasing in Asia, including Japan. Although this trend has been attributed partly to a more Western diet, this assumption may involve variable confounders. Thus, we examined the histological features of contemporary vs historical latent prostate cancer. MATERIALS AND METHODS: Prostate specimens from a consecutive autopsy series (127, present study, 2008 to 2013) were examined. Each prostate gland was fixed and sliced in step sections. The findings were compared to those from another autopsy series (501 subjects, 1983 to 1987) at our institution. RESULTS: The mean age of subjects in the present study was 68.9 years while the mean age was not available from the earlier study. However, the mean age of the 566 entrants in the expanded database (1983 to 1989) was 63.5 years (p=0.0001). Prostate weight was significantly greater in the present study (p <0.0001). Latent prostate cancer was found more frequently in the present study than in the previous study (43.3% and 20.8%, respectively, p <0.0001). No distinct difference was seen in the proportion of tumor grade between the groups. An increasing trend of moderately to poorly differentiated tumors with advancing age was more evident in the present study. Index cancer volume was greater in the present study with 25.5% measuring 500 mm(3) or greater vs only 9.6% of cancers in the previous study (p=0.008). CONCLUSIONS: Chronological changes in the histological characteristics of Japanese latent prostate cancer were noted as it is more prevalent in the contemporary series. Our data may reflect a worldwide trend in increasingly aging societies.

Antipyretic analgesic drugs have different mechanisms for regulation of the expression of inducible nitric oxide synthase in hepatocytes and macrophages.

Antipyretic analgesic drugs (including non-steroidal anti-inflammatory drugs) inhibit cyclooxygenase-2 and inducible nitric oxide synthase (iNOS), resulting in decreases of the proinflammatory mediators prostaglandin E2 and nitric oxide (NO), respectively. Both mediators are regulated by nuclear factor-kappa B (NF-κB), a key transcription factor in inflammation. Few reports have compared the efficacy and potency of anti-inflammatory drugs as NO inhibitors. In our study, we examined the effects of four popular antipyretic analgesic drugs on NO production induced in hepatocytes and macrophages. Mouse RAW264.7 macrophages treated with bacterial lipopolysaccharide showed the highest efficacy with regard to NO production; aspirin, loxoprofen, ibuprofen, and acetaminophen dose-dependently suppressed NO induction. Ibuprofen showed the highest potency in suppressing the induced production of NO. In rat hepatocytes, all the drugs inhibited interleukin 1ß-induced NO production and ibuprofen and loxoprofen inhibited NO induction effectively. Unexpectedly, the potency of NO suppression of each drug in hepatocytes did not always correlate with that observed in RAW264.7 cells. Microarray analyses of mRNA expression in hepatocytes revealed that the effects of the four antipyretic analgesic drugs modulated the NF-κB signaling pathway in a similar manner to the regulation of the expression of genes associated with inflammation, including the iNOS gene. However, the affected signal-transducing molecules in the NF-κB pathway were different for each drug. Therefore, antipyretic analgesic drugs may decrease NO production by modulating the NF-κB pathway in different ways, which could confer different efficacies and potencies with regard to their anti-inflammatory effects.

Interleukin-1ß induces tumor necrosis factor-α secretion from rat hepatocytes.

AIM: Tumor necrosis factor-α (TNF-α) is a pleiotropic cytokine involved in various inflammatory diseases. The only production of TNF-α in the liver is thought to be from hepatic macrophages known as Kupffer cells, predominantly in response to bacterial lipopolysaccharide (LPS). METHODS: Primary cultured rat hepatocytes were used to analyze TNF-α expression in response to the pro-inflammatory cytokine, interleukin-1ß (IL-1ß). Livers of rats subjected to LPS-induced endotoxemia were analyzed. RESULTS: Immunocytochemistry and enzyme-linked immunosorbent assays demonstrated that IL-1ß-treated rat hepatocytes secreted TNF-α, and RNA analyses indicated that TNF-α mRNA was induced specifically by IL-1ß. Northern blot analysis showed that not only mRNA, but also a natural antisense transcript (asRNA), was transcribed from the rat Tnf gene in IL-1ß-treated hepatocytes. TNF-α was detected in the hepatocytes of LPS-treated rats. Both TNF-α mRNA and asRNA were expressed in the hepatocytes of LPS-treated rats, human hepatocellular carcinoma and human monocyte/macrophage cells. To disrupt the interaction between TNF-α asRNA and TNF-α mRNA, sense oligonucleotides corresponding to TNF-α mRNA were introduced into rat hepatocytes resulting in significantly increased levels of TNF-α mRNA. One of these sense oligonucleotides increased a half-life of TNF-α mRNA, suggesting that the TNF-α asRNA may reduce the stability of TNF-α mRNA. CONCLUSION: IL-1ß-stimulated rat hepatocytes are a newly identified source of TNF-α in the liver. TNF-α mRNA and asRNA are expressed in rats and humans, and the TNF-α asRNA reduces the stability of the TNF-α mRNA. Hepatocytes and TNF-α asRNA may be therapeutic targets to regulate levels of TNF-α mRNA.

[Two cases of anaplastic carcinoma of the pancreas in terms of the diversity of carcinoma cells (epithelial-mesenchymal transition; EMT)].

The first evaluated case was a 71-year-old man with melaena. A huge tumor mass was observed in the tale of the pancreas, which we diagnosed as pancreatic carcinoma. Pathological findings confirmed that it was anaplastic pancreas carcinoma (giant cell type). The second case was a 61-year-old woman with upper abdominal pain. Irregular tumors with scattered cysts were observed in the tale of the pancreas, which rapidly increased. Autopsy findings confirmed that it was anaplastic pancreas carcinoma (pleomorphic type). Both cases showed positive for epithelial marker and mesenchymal markers and decrease in stainability by E-cadherin staining, a result which suggested diversity of tumor cells in anaplastic pancreas carcinomas.

Comparative examination of anti-proliferative activities of (-)-epigallocatechin gallate and (--)-epigallocatechin against HCT116 colorectal carcinoma cells.

We compared anti-proliferative activities of (-)-epigallocatechin gallate (EGCG) and (-)-epigallocatechin (EGC) against HCT116 colorectal carcinoma cells. These catechins inhibited cell growth to nearly the same extent at low cell confluency in plates. However, their inhibitory effect grew weaker as cell confluence increased, and this tendency was more conspicuous for EGC than for EGCG. Both EGCG and EGC activated the phosphorylation of the major MAPKs, ERK, JNK, and p38, in the HCT116 cells as in many other established human cancer cells though to different extents. Cell cycle analyses, DNA fragmentation assays, and TUNEL assays as well as Western blot assays suggested that these catechins inhibited cell growth through mitogen-activated protein kinase (MAPK)-mediated apoptosis rather than cell cycle regulation.

Safer endoscopic gastric mucosal resection: preoperative proton pump inhibitor administration.

BACKGROUND AND AIM: Control of bleeding is crucial in improving the safety of endoscopic mucosal resection (EMR), and intragastric acidity has a great impact on hemostasis and blood coagulation. Proton pump inhibitors (PPI) are potent suppressors of gastric acid; however, PPI need to be continuously administered orally for several days, and thus initial effects may be insufficient if PPI is only administered immediately after EMR. The aim of this study was to determine whether preoperative administration of PPI prior to EMR can elevate intragastric acidity, facilitate better control of intraoperative bleeding (complete coagulation and hemostasis), prevent postoperative bleeding, and facilitate healing of artificial ulcers. METHODS: A randomized clinical study was conducted in which EMR was performed with or without 1 week of preoperative PPI administration. RESULTS: Artificial ulcers created by EMR healed more rapidly in patients who received preoperative PPI. CONCLUSIONS: The results of the study suggest that preoperative administration of PPI before EMR is useful for controlling and preventing bleeding, and for facilitating the healing of artificial ulcers.

[Pilot study of concomitant TS-1/CDGP radiotherapy for esophageal cancer].

Although surgery is treatment of choice for esophageal cancer, radiochemotherapy is being employed throughout Japan for the purpose of improving patient QOL. The results of this therapy are reported to be comparable to those associated with surgical treatment. However, since concomitant 5-FU/CDDP radiotherapy, currently the treatment of choice when implementing radiochemotherapy, is associated with a comparatively high incidence of gastrointestinal disorders and requires continuous intravenous infusion for 24 hours, it lowers the level of patient QOL. We have proposed a clinical study of concomitant TS-1/CDGP radiotherapy for the purpose of maintaining patient QOL and improving outcome. We conducted a pilot study prior to the phase I and II studies. The study was conducted on six cases and favorable results were obtained, consisting of a CR rate of 66.7% and a two-year survival rate of 50%. Although bone marrow inhibition was observed as an adverse side effect, gastrointestinal disorders that were discernible to the patients were extremely mild, and patient QOL was able to be maintained. CR was observed in 2 cases who were positive for DPD as determined by immunostaining. We are planning on conducting phase I and II studies in the future based on the potential for this treatment to contribute to the preservation of patient QOL and improve prognosis.

[Clinical study of the combination of small amount of nedaplatin (CDGP)/5-FU with radiation for the treatment of esophageal cancer].

Recently chemoradiotherapy for esophageal cancer has been drawing public attention to the issue of quality of life maintenance for patients. Although the standard method of chemoradiotherapy is CDDP/5FU, it has been claimed that CDGP (a derivative of CDDP) alone is more effective than CDDP for the treatment of esophageal cancer due to its low nephro- and digestive toxicity. We used a small amount of CDGP/5-FU in combination with radiation instead of CDDP, for the treatment of esophageal cancer and performed clinical examination of patients. The partial response rate was 80% and the complete response rate was 50%. Major side-effects were leukopenia, neutropenia, thrombocytopenia and anemia. Further study of dosage and schedule is necessary, however, CDGP/5-FU combined with radiation therapy could be used as choices of chemoradiotherapy for esophageal cancer in the future.