A Genome-Wide Association Study of Prostate Cancer Susceptibility Using Occupational and Environmental Factors as Confounding Factors.

Background In addition to genetic predisposition, occupational and environmental factors are important for the risk of prostate cancer. We investigated the effect of single nucleotide polymorphisms (SNPs) on the development of prostate cancer in Japan, including occupational and industrial history as confounding factors in addition to age, smoking, and alcohol drinking. Methods We enrolled 210 prostate cancer patients and 504 male control patients. We conducted four genome-wide association study (GWAS) patterns for prostate cancer development. In the association test, logistic regression models incorporated age, smoking history, alcohol consumption history, and each pattern of industrial/occupational classification. Results No SNPs satisfying the genome-wide significance level of 5×10-8 were detected in GWAS. SNPs with a suggestive association level of 1×10-6 were found near the long intergenic non-protein coding RNA 1824 (LINC01824) and tripartite motif family like 2 (TRIML2) genes in the GWAS using occupational history as a confounder and near the ribosomal protein S2 pseudogene 25 (RPS2P25) gene in the GWAS using industrial history as a confounder. No SNPs that met the suggestive association level were observed in the GWAS that did not include occupational and industrial history. Conclusion By adding occupational and industrial history to the confounding factors, there were SNPs detected in the GWAS for prostate cancer development. The consideration of occupational and industrial history may increase the usefulness of GWAS.

A radiation-related second primary gastrointestinal stromal tumor in the bladder.

A 69-year-old man underwent 78 Gy/39 Fr of intensity-modulated radiation therapy for prostate cancer. Seven years after radiotherapy, a nonpapillary bladder tumor was identified. Transurethral resection of the bladder tumor was performed, and the pathological diagnosis was spindle cell sarcoma. Immunostaining revealed KIT-, DOG1++, CD34-, Actin++, Cytokeratin-, Desmin-, S100 protein-, and Vimentin++. No tumor recurrence was observed until 17 months after tumor resection. DOG1 is strongly and specifically expressed in gastrointestinal stromal tumors. This was a rare case of bladder gastrointestinal stromal tumor as a radiation-related second primary tumor.

Hypomethylation of a CpG Site in the CpG Island of the Aquaporin 1 Gene May Be Involved in the Formation of Adult-Onset Non-communicating Hydrocele Testis.

Background Water channel aquaporin 1 (AQP1) protein expression is enhanced in the tunica vaginalis of patients with adult-onset non-communicating hydrocele testis and may contribute to the development of non-communicating hydrocele testis. We performed genetic and epigenetic analyses of the AQP1 gene in the tunica vaginalis of patients with adult-onset non-communicating hydrocele testis to elucidate the cause of enhanced AQP1 protein expression. Methodology The genotype was determined for Tag single-nucleotide polymorphisms (SNPs) representing the AQP1 gene and SNPs in the 5'-upstream region of the AQP1 gene. Then, by performing association analysis, the applicability of various genetic models was investigated for each SNP. Moreover, the methylation rate of CpG sites was examined for the CpG island related to the AQP1 gene. Results There was no significant association between each SNP and hydrocele testis for any of the genetic models. The average methylation rate of the 17 CpG sites evaluated was not significantly different between controls and hydrocele testis, but the methylation rate was lower in hydrocele testis than in controls at one CpG site. Conclusions There was a significant decrease in the methylation rate at one of the CpG sites in the CpG island associated with the AQP1 gene in the tunica vaginalis of patients with non-communicating hydrocele testis. This may increase AQP1 protein expression and contribute to the formation of hydrocele testis. SNPs related to the AQP1 gene were not associated with hydrocele testis.

Structural Analysis of the Colony-Stimulating Factor 3 Gene of Granulocyte Colony-Stimulating Factor-Producing Urothelial Cancer.

Background Granulocyte colony-stimulating factor (G-CSF) is a member of the CSF family of glycoproteins that regulate the proliferation, differentiation, and mobilization of neutrophils. G-CSF-producing malignant cancers have been reported to occur in various organs and are mostly associated with poor clinical prognosis. Here, we analyzed the structure of the CSF3 gene encoding the G-CSF protein to delineate the mechanism of G-CSF production by the cancer cells. Methodology Two cases of G-CSF-producing urothelial cancers and three cases of G-CSF-nonproducing bladder cancers were enrolled for genetic analysis. Results In one case of G-CSF-producing bladder cancer, six somatic mutations were detected in the 5'- upstream region of the CSF3 gene. No somatic mutations in the CSF3 gene were detected in another case of G-CSF-producing renal pelvic cancer and G-CSF-nonproducing bladder cancers. Copy numbers of the CSF3 gene were not increased in G-CSF-producing urothelial cancers. Conclusions Somatic mutations in the 5'- upstream region of the CSF3 gene may cause G-CSF protein overproduction.

Relationship between the Reduced Expression of Zinc Finger Protein 668 in Bladder Cancer and Its Invasiveness.

The zinc finger protein 668 (ZNF668) gene encodes a Kruppel C2H2-type zinc-finger protein with 16 C2H2-type zinc fingers. The ZNF668 gene functions as a tumor suppressor gene in breast cancer. We histologically analyzed ZNF668 protein expression in bladder cancer and examined mutations of the ZNF668 gene in 68 cases of bladder cancer. In bladder cancer, the ZNF668 protein was expressed in the nuclei of cancer cells. In bladder cancer with submucosal and muscular infiltration, the expression of ZNF668 protein was significantly lower than that without submucosal and muscular infiltration. Eight heterozygous somatic mutations were detected in exon3 in five cases, and five of the mutations resulted in amino acid sequence mutations. Mutations resulting in amino acid sequence alterations also resulted in lower ZNF668 protein expression in bladder cancer cell nuclei, but no significant association with bladder cancer infiltration was detected. Decreased ZNF668 expression in bladder cancer was associated with submucosal and muscle invasion of cancer cells. Somatic mutations resulting in amino acid mutations in ZNF668 were found in 7.3% of the bladder cancer cases.

Genome-Wide Association Study Adjusted for Occupational and Environmental Factors for Bladder Cancer Susceptibility.

This study examined the effects of single-nucleotide polymorphisms (SNPs) on the development of bladder cancer, adding longest-held occupational and industrial history as regulators. The genome purified from blood was genotyped, followed by SNP imputation. In the genome-wide association study (GWAS), several patterns of industrial/occupational classifications were added to logistic regression models. The association test between bladder cancer development and the calculated genetic score for each gene region was evaluated (gene-wise analysis). In the GWAS and gene-wise analysis, the gliomedin gene satisfied both suggestive association levels of 10-5 in the GWAS and 10-4 in the gene-wise analysis for male bladder cancer. The expression of the gliomedin protein in the nucleus of bladder cancer cells decreased in cancers with a tendency to infiltrate and those with strong cell atypia. It is hypothesized that gliomedin is involved in the development of bladder cancer.

Selection of AVP-Shortage Patients as Candidates for Low-Dose Oral Desmopressin Administration.

OBJECTIVE: We herein attempted to select male patients with an elevated nocturnal urinary frequency possibly due to a shortage of AVP. These patients may be good candidates for low-dose oral desmopressin administration. PATIENTS AND METHODS: Serum and spot urine osmolality, electrolytes, serum creatinine, casual blood glucose, plasma brain natriuretic polypeptide (BNP), and plasma AVP were measured at the same time in 97 elderly male patients with urinary symptoms under free water drinking. RESULTS: A binary plot of plasma AVP and serum osmolality indicated a region at which patients had relatively lower AVP considering higher serum osmolality. It was tentatively named the desmopressin region. Twenty out of 97 (20.6%) patients were in the desmopressin region. Daily urine output did not exceed 3 L in any patient. Urine osmolality was slightly lower in patients in the desmopressin region. No significant differences were observed in urine volume, urinary frequency, or urination questionnaire scores between both groups. CONCLUSION: AVP-shortage patients may be selected for treatment with oral desmopressin based on measurements of serum osmolality and plasma AVP.

[Renal Cancer Tissue after Nivolumab/Ipilimumab Combination Therapy for Metastatic Renal Cell Carcinoma].

Antibodies that inhibit the function of PD-1, PD-L1, and CTLA4 increase the tumor immune response and suppress tumor growth. These immune checkpoint inhibitors have also been introduced into the treatment of metastatic renal cancer. We report combination therapywith nivolumab and ipilimumab in a case of renal cancer with bone metastasis and subsequent removal of the primarytumor. The patient was a 67-year-old male. Computed tomography (CT) revealed a 6.5×5.6 cm renal tumor in the left kidney, with osteolytic metastasis to the left 11th rib and thoracic spine. Irradiation of 30 Gy was performed for the thoracic spine tumors. Then, nivolumab+ipilimumab combination therapywas continued for a total of 4 courses. In addition, 4 courses of nivolumab monotherapywere added. CT after therapyrevealed a 15. 4% decrease in the length-wise diameter and increase in internal necrosis in the left kidneytumor, as well as shrinkage, absorption reduction, and appearance of a hardened border in the bone metastasis. Therefore, transabdominal left nephrectomywas performed. The histopathological diagnosis was clear cell carcinoma pT1bN0, grade 2. The renal cancer tissue was mostlyinternal necrosis, and the viable tumor comprised approximately10%. Marked infiltration of predominantlyCD8-positive lymphocytes to the primarytumor site was observed. Postoperatively, nivolumab was discontinued and the patient is under observation. In the specimens from 2 patients treated using pazopanib, a tyrosine kinase inhibitor, prior to renal cell cancer removal, there were fewer CD8- and CD4-positive cells infiltrating the renal cancer than after the combination therapy.

[A Case of Unclassified Sex Cord-Stromal Tumor Containing hCG-Positive Cells].

We present a case of unclassified sex cord-stromal testicular tumor with lung metastasis. A 48-year-old man consulted our hospital for left testicular enlargement that began 3 years ago. Computed tomography revealed a heterogeneously enhanced left testicular tumor 11 cm in diameter and a 5 mm metastatic lung tumor. The human chorionic gonadotropin (hCG) level was elevated (141.6 mU/ml), whereas the levels of α-fetoprotein (AFP) and LDH were normal. The external genitalia were normal and gynecomastia was not observed. Left high orchiectomy followed by 3 cycles of BEP chemotherapy (bleomycin, etoposide, and cisplatin) every 4 weeks was performed. The pathology of the excised specimen was unclassified sex cordstromal testicular tumor containing hCG-positive cells. On immunohistochemistry, the tumor cells were partly positive for AE1/AE3, hCG, and calretinin. Vimentin was diffusely positive, but OCT3/4, SALL4, GATA3, and CK7 were negative. After BEP treatment, the metastatic lung lesion disappeared. Unclassified sex cord-stromal testicular tumor is a rare disease and its treatment has not been established. Thus, further accumulation of cases is needed.

Prediction of prostate cancer by deep learning with multilayer artificial neural network.

INTRODUCTION: To predict the rate of prostate cancer detection on prostate biopsy more accurately, the performance of deep learning using a multilayer artificial neural network was investigated. METHODS: A total of 334 patients who underwent multiparametric magnetic resonance imaging before ultrasonography guided transrectal 12-core prostate biopsy were enrolled in the analysis. Twenty-two non-selected variables, as well as selected ones by least absolute shrinkage and selection operator (Lasso) regression analysis and by stepwise logistic regression analysis, were input into the constructed multilayer artificial neural network (ANN) programs; 232 patients were used as training cases of ANN programs and the remaining 102 patients were for the test to output the probability of prostate cancer existence, accuracy of prostate cancer prediction, and area under the receiver operating characteristic (ROC) curve with the learned model. RESULTS: With any prostate cancer objective variable, Lasso and stepwise regression analyses selected 12 and nine explanatory variables, respectively, from 22. Using trained ANNs with multiple hidden layers, the accuracy of predicting any prostate cancer in test samples was about 5-10% higher compared to that with logistic regression analysis (LR). The area under the curves (AUC) with multilayer ANN were significantly larger on inputting variables that were selected by the stepwise LR compared with the AUC with LR. The ANN had a higher net benefit than LR between prostate cancer probability cutoff values of 0.38 and 0.6. CONCLUSIONS: ANN accurately predicted prostate cancer without biopsy marginally better than LR. However, for clinical application, ANN performance may still need improvement.

A Case of Idiopathic Spontaneous Intratesticular Hemorrhage.

A 21-year-old man presented with acute-onset left lower abdominal pain that had initially developed 8 hours earlier. He was not given any medication including anti-coagulants. He denied trauma. On palpation, the left testis was slightly swollen and showed tenderness. The suspected diagnosis was testicular torsion, and surgical exploration was indicated. On visual inspection, the whole testis was black. The spermatic cord was neither distorted nor black. Testicular torsion could not be completely ruled out; thus, left orchiectomy was performed. Histopathology revealed diffuse intratesticular hemorrhage without the necrosis of seminiferous tubular cells. We encountered a case of idiopathic spontaneous intratesticular hemorrhage.

A single nucleotide polymorphism in kidney anion exchanger 1 gene is associated with incomplete type 1 renal tubular acidosis.

Various conditions including distal renal tubular acidosis (dRTA) can induce stone formation in the kidney. dRTA is characterized by an impairment of urine acidification in the distal nephron. dRTA is caused by variations in genes functioning in intercalated cells including SLC4A1/AE1/Band3 transcribing two kinds of mRNAs encoding the Cl-/HCO3- exchanger in erythrocytes and that expressed in α-intercalated cells (kAE1). With the acid-loading test, 25% of urolithiasis patients were diagnosed with incomplete dRTA. In erythroid intron 3 containing the promoter region of kAE1, rs999716 SNP showed a significantly higher minor allele A frequency in incomplete dRTA compared with non-dRTA patients. The promoter regions of the kAE1 gene with the minor allele A at rs999716 downstream of the TATA box showed reduced promoter activities compared that with the major allele G. Patients with the A allele at rs999716 may express less kAE1 mRNA and protein in the intercalated cells, developing incomplete dRTA.

Oct4B, CD90, and CD73 are upregulated in bladder tissue following electro-resection of the bladder.

AIM: We tested the hypothesis that stimulation by electro-resection of bladder tissue induces stem cells in the tissue repair process. MATERIALS & METHODS: After primary transurethral resection of a bladder tumor and surrounding tissue (TUR-Bt), second TUR-Bt was performed. Tissues excised by second TUR-Bt were immunohistochemically stained for Oct4, a marker of pluripotency, and for CD90 and CD73, markers of mesenchymal stromal cells, when no bladder tumor cells remained. RESULTS AND CONCLUSIONS: Oct4B protein was sporadically stained in the cytoplasm of interstitial cells in four out of eight cases. CD90 and CD73 are upregulated in interstitial and vascular endothelial cells without CD45 expression. Mesenchymal stromal cells, but not pluripotent stem cells, may be mainly involved in bladder tissue repair.

Detection of AR-V7 mRNA in whole blood may not predict the effectiveness of novel endocrine drugs for castration-resistant prostate cancer.

A splice variant of androgen receptor (AR), AR-V7, lacks in androgen-binding portion and leads to aggressive cancer characteristics. Reverse transcription-polymerase chain reactions (PCRs) and subsequent nested PCRs for the amplification of AR-V7 and prostate-specific antigen (PSA) transcripts were done for whole blood of patients with prostate cancer and male controls. With primary reverse transcription PCRs, AR-V7 and PSA were detected in 4.5% and 4.7% of prostate cancer, respectively. With nested PCRs, AR-V7 messenger RNA (mRNA) was positive in 43.8% of castration-sensitive prostate cancer and 48.1% of castration-resistant prostate cancer (CRPC), while PSA mRNA was positive in 6.3% of castration-sensitive prostate cancer and 18.5% of CRPC. Whole-blood samples of controls showed AR-V7 mRNA expression by nested PCR. Based on multivariate analysis, expression of AR-V7 mRNA in whole blood was not significantly correlated with clinical parameters and PSA mRNA in blood, while univariate analysis showed a correlation between AR-V7 mRNA and metastasis at initial diagnosis. Detection of AR-V7 mRNA did not predict the reduction of serum PSA in patients with CRPC following abiraterone and enzalutamide administration. In conclusion, AR-V7 mRNA expression in normal hematopoietic cells may have annihilated the manifestation of aggressiveness of prostate cancer and the prediction of the effectiveness of abiraterone and enzalutamide by the assessment of AR-V7 mRNA in blood.

Development of nephrotic syndrome after administration of sorafenib in a case of metastatic renal cell carcinoma.

Nephrotic syndrome, after administration of tyrosine kinase inhibitors, is uncommon and not well known. A 62-year-old male, who had experienced a left nephrectomy due to a traffic accident 38 years ago, underwent a partial nephrectomy for right renal cell carcinoma (RCC). Histologically, the tumor was a clear cell RCC. Two years later abdominal CT revealed para-aortic lymph node metastasis. During these two years, serum creatinine had increased from 2.0 mg/dL to 2.9 mg/dL along with the appearance of proteinuria. After only a week of sorafenib, 400 mg/day, fever developed and sorafenib was stopped. Although normotensive, his serum creatinine increased to 3.83 mg/dL and serum albumin decreased from 1.8 g/L to 1.0 g/L. Proteinuria also worsened to 27.5 g/day. He became edematous, and ascites and cardiac effusions also appeared. He was diagnosed with nephrotic syndrome. A retrospective review of the histology of the partial nephrectomy revealed no change in the glomeruli.