Impact of Tumor Grade Distribution on Genetic Alterations in Clear Cell Renal Cell Carcinoma and Prostate Cancer.

BACKGROUND/AIM: A genomic analysis based on next-generation sequencing is important for deciding cancer treatment strategies. Cancer tissue sometimes displays intratumor heterogeneity and a pathologic specimen may contain more than two tumor grades. Although tumor grades are very important for the cancer prognosis, the impact of higher tumor grade distribution in a specimen used for a genomic analysis is unknown. PATIENTS AND METHODS: We retrospectively analyzed the data of 61 clear cell carcinoma and 46 prostate cancer patients that were diagnosed between December 2018 and August 2022 using the GeneRead Human Comprehensive Cancer Panel or SureSelect PrePool custom Tier2. Genome annotation and curation were performed using the GenomeJack software. RESULTS: Tumor mutation burden (TMB) was increased in proportion to the higher tumor grade distribution in grade 2 clear cell renal cell carcinoma (ccRCC). In PC, Grade Group 3/4 specimens that included an increased distribution of Gleason pattern 4 had more frequent gene mutations. CONCLUSION: Our results suggest the importance of selecting the maximum distribution of higher tumor grade areas to obtain results on the precise gene alterations for genomics-focused treatments.

Derivative Chromosome 3 Loss from t(3;6)(q12;q14) Followed by Differential VHL Mutations Underlie Multifocal ccRCC.

BACKGROUND/AIM: The Von Hippel-Lindau (VHL) gene encodes a protein (pVHL) that plays an important role in proteasome degradation of hypoxia inducible factor α (HIFα) through E3 activation. Accumulation of HIFα by loss of functional pVHL promotes tumorigenesis, thus, VHL has tumor suppressor gene capability in clear cell renal cell carcinoma (ccRCC). VHL is the most frequently mutated gene in ccRCC. The complete loss of VHL is mainly achieved by loss of chromosome 3p, which has a VHL coding region in combination with mutation or hypermethylation of the remaining copy of VHL. Given the risk of constitutional chromosome 3 translocation for RCC, it is important to detect the translocation and understand the mechanism underlying the development of multifocal ccRCC. CASE REPORT: A 67-year-old female patient diagnosed with multifocal RCC underwent robot-assisted partial nephrectomy (RAPN) for three kidney tumors. A cancer gene panel test using next generation sequencing (NGS) detected differential VHL mutations (c.533T>G; p.L178R, c.465_466insTA; p.T157Ifs*3, c.343C>A; p.H115N), while VHL mutation was not detected in peripheral blood DNA. A tendency toward copy number loss of genes on der(3) was also detected in all tumors, but not in the germline one. A karyotype analysis revealed a germline translocation between 3 and 6, t(3;6)(q12;q14). CONCLUSION: Chromosome 3 translocation and loss of derivative chromosome containing 3p and subsequent somatic differential VHL mutations in this case strongly support the previously proposed three-step model to explain the development of familial conventional ccRCC.

The feasibility of dedicated breast PET for the assessment of residual tumor after neoadjuvant chemotherapy.

PURPOSE: To evaluate the utility of ring-type dedicated breast positron emission tomography (dbPET) for the detection of the residual tumor after neoadjuvant chemotherapy (NAC). MATERIALS AND METHODS: This prospective study included 27 women with histologically proven breast cancer over a 37-month period. All patients underwent ring-type dbPET followed by whole-body PET-CT (WBPET) for preoperative tumor evaluation and re-staging after NAC. The maximum standardized uptake value (SUVmax) of the tumor lesion and the degree of confidence for the presence of the residual tumor were compared between pathological complete response (pCR) and non-pCR tumors. The sensitivity, specificity, and area under the receiver operating characteristic curve (AUC) for the detection of a non-pCR tumor were compared between dbPET and WBPET. RESULTS: On dbPET, SUVmax was significantly higher in non-pCR than in pCR tumors (P = 0.030). The sensitivity for the detection of a non-pCR tumor was significantly higher with dbPET than with WBPET (84.2% vs 26.3%, P = 0.001). In the qualitative analysis, the sensitivity for the detection of a non-pCR tumor was also significantly higher with dbPET than with WBPET (57.9% vs 21.1%, P = 0.016). CONCLUSION: The dbPET can provide more sensitive detection of residual tumor after NAC than can WBPET.

18-F fluorodeoxyglucose uptake in positron emission tomography as a pathological grade predictor for renal clear cell carcinomas.

OBJECTIVES: To evaluate the usefulness of Fluorine-18 fluorodeoxyglucose positron emission tomography/computed tomography (18-F FDG-PET/CT) in the prediction of Fuhrman pathological grades of renal clear cell carcinoma (cRCC). METHODS: This retrospective study was approved by our institutional review board, and written informed consent was waived. Thirty-one patients with pathologically proven cRCC underwent 18-F FDG-PET/CT for tumour staging. Maximum standardized uptake value of cRCC (tumour SUVmax) and mean SUV of the liver and spleen (liver and spleen SUVmean) were measured by two independent observers. Tumour SUVmax, tumour-to-liver SUV ratio, and tumour-to-spleen SUV ratio were correlated with the pathological grades. RESULTS: Logistic analysis demonstrated that only the tumour-to-liver SUV ratio was a significant parameter for differentiating high-grade (Fuhrman grades 3 and 4) tumours from low-grade (Fuhrman grades 1 and 2) tumours (P = 0.007 and 0.010 for observers 1 and 2, respectively). Sensitivity, specificity, and positive and negative predictive values for detecting tumours of Fuhrman grades 3 and 4 were 64, 100, 100, and 77%, respectively, for observer 1, and 79, 88, 85, and 83%, respectively, for observer 2. CONCLUSIONS: The tumour-to-liver SUV ratio with 18-F FDG-PET/CT appeared to be a valuable imaging biomarker in the prediction of high-grade cRCC. KEY POINTS: • Tumour SUV max was correlated with the Fuhrman grades. • High-grade tumours have significantly higher SUV max than low-grade tumours. • Tumour-to-liver SUV ratio is useful in the prediction of high-grade cRCC.

Molecular diversity of eukaryotes in municipal wastewater treatment processes as revealed by 18S rRNA gene analysis.

Eukaryotic communities involved in sewage treatment processes have been investigated by morphological identification, but have not yet been well-characterized using molecular approaches. In the present study, eukaryotic communities were characterized by constructing 18S rRNA gene clone libraries. The phylogenetic affiliations of a total of 843 clones were Alveolata, Fungi, Rhizaria, Euglenozoa, Stramenopiles, Amoebozoa, and Viridiplantae as protozoans and Rotifera, Gastrotricha, and Nematoda as metazoans. Sixty percent of the clones had <97% sequence identity to described eukaryotes, indicating the greater diversity of eukaryotes than previously recognized. A core OTU closely related to Epistylis chrysemydis was identified, and several OTUs were shared by 4-8 libraries. Members of the uncultured lineage LKM11 in Cryptomycota were predominant fungi in sewage treatment processes. This comparative study represents an initial step in furthering understanding of the diversity and role of eukaryotes in sewage treatment processes.

Microbial community composition of a down-flow hanging sponge (DHS) reactor combined with an up-flow anaerobic sludge blanket (UASB) reactor for the treatment of municipal sewage.

The microbial community composition of a down-flow hanging sponge (DHS) reactor in an up-flow anaerobic sludge blanket (UASB)-DHS system used for the treatment of municipal sewage was investigated. The clone libraries showed marked differences in microbial community composition at different reactor heights and in different seasons. The dominant phylotypes residing in the upper part of the reactor were likely responsible for removing organic matters because a significant reduction in organic matter in the upper part was observed. Quantification of the amoA genes revealed that the proportions of ammonia oxidizing bacteria (AOB) varied along the vertical length of the reactor, with more AOB colonizing the middle and lower parts of the reactor than the top of the reactor. The findings indicated that sewage treatment was achieved by a separation of microbial habitats responsible for organic matter removal and nitrification in the DHS reactor.

Characterization of the retained sludge in a down-flow hanging sponge (DHS) reactor with emphasis on its low excess sludge production.

Experiments to characterize retained sludge in a down-flow hanging sponge (DHS) reactor fed with upflow anaerobic sludge blanket (UASB) treated sewage under moderate conditions were conducted. Plenty of oxygen was supplied through the DHS reactor without aeration and the effluent qualities after the reactor were comparable to activated sludge processes. The average excess sludge production rate was 0.09 g SS g(-1) COD removed. The DHS reactor maintained a high sludge concentration of 26.9 g VSS L(-1) sponge, resulting in a low loading rate of 0.032 g COD g(-1) VSS day(-1). The endogenous respiration rate of DHS sludge was comparable to previously reported aerobic sludges. The numbers of microfauna were one order of magnitude greater than those in activated sludge. The results indicated that low excess sludge production was attributable to the high sludge concentration, sufficient oxygen supply, adequate endogenous respiration rate, and a high density and diversity of microfauna.

A case of malignant melanoma simultaneously found in the lung and small intestine.

Metastatic melanoma in the small bowel is a common cause of secondary intestinal tumors. We present a case of a 77-year-old man with melena resulting from melanoma in the small bowel that was simultaneously found with malignant melanoma in the lung. Abdominal contrast-enhanced computed tomography (CT) and position emission tomography (PET) revealed a 30 × 20 mm hypervascular lesion in the small bowel and a 9 × 9 mm right lobe lung mass with metastasis in the mediastinal and peritoneal lymph nodes, and the spleen. The bowel and lung tumor lesions were confirmed by enteroscopy and bronchoscopy, respectively, and were histologically diagnosed as malignant melanoma. In cases of small bowel malignant melanoma, an extraintestinal origin should always be suspected because intestinal melanomas are almost always metastatic. PET/CT is an effective tool for confirming intestinal melanomas because of its high sensitivity and specificity.

Combined adenomatoid tumor and well differentiated papillary mesothelioma of the omentum.

Herein is reported a highly rare case of combined adenomatoid tumor and well differentiated papillary mesothelioma of the omentum. A 45-year-old Japanese man was incidentally found to have a mesenteric mass during abdominal ultrasonography. Grossly, a well-circumscribed and non-encapsulated tumor, measuring 24 × 23 × 22 mm, was located in the omentum with no obvious invasion to the surrounding stomach and pancreas. Microscopically, the tumor consisted of two histological distinct components; the majority of it showed irregular arrangement of numerous cysts lined by a single layer of flattened or epithelioid cells and the multifocal minor component exhibited prominent papillary protrusions lined by a single layer of cuboidal cells with relatively uniform nucleus. Immunohistochemically, the tumor cells were diffusely positive for cytokeratin (AE1/AE3), vimentin, calretinin, D2-40 and WT-1, and negative for epithelial membrane antigen (EMA), desmin, S-100 protein and CD68. Ki-67 labeling index of the tumor cells was <5%. The postoperative course was uneventful, though he has not received any adjuvant therapy. Despite the rarity, recognition of such combined cases of mesothelial lesions is important to prevent overdiagnosis.

[A case of well differentiated mucinous carcinoma of the gallbladder].

We report a case of well differentiated mucinous carcinoma of the gallbladder. An 80-year-old man was admitted to our hospital because of fever and CT showed a mass in the fundus of the gallbladder and choledocholithiasis. Endoscopic removal of the common bile duct stones was done. Abdominal ultrasonography and CT showed a 4 cm mass in the fundus of the gallbladder. Based on a diagnosis of adenomyomatosis or mucinous carcinoma of the gallbladder, a laparotomy revealed a white mass in the fundus of gallbladder. Intra-operative pathological diagnosis of lymph node of cystic duct was mucinous carcinoma. Therefore, we conducted partial resection of the liver with D1 lymph node dissection. Pathological diagnosis confirmed well differentiated mucinous carcinoma of the gallbladder. His postoperative course was uneventful, and he is doing well without recurrence.

[Present status and prospects about an animal model of post-traumatic stress disorder in rats using a shuttle box].

Wistar rats exposed to inescapable foot shocks (IS) for 2 wk exhibited PTSD-like bi-directional changes similar to avoidance/numbing and hyperarousal symptoms when placed in a shuttle box. Paroxetine administration after IS reduced the hyperarousal-like behavior, and its therapeutic effect on avoidance/numbing-like behavior was also significant. Further, F344 rats, which were more vulnerable to various kinds of stressors, showed more significant 'bi-directional changes' than Wistar rats. Thus, the paradigm we have developed could serve as a useful PTSD model because of its face, predictive, and construct validity. Moreover, the intensity of IS dose-dependently induced PTSD-like behaviors and hypo-activity in a shuttle box, similar to the 'avoidance/numbing' that reappeared in a square open field. These findings further support the construct validity of this paradigm. Both electro-convulsive shock treatment before and after IS ameliorated the PTSD-like behaviors in this model, so electro-convulsive therapy may be an effective method for prevention and medical treatment of PTSD in the future. On the other hand, pretreatment with fluvoxamine before IS did not have a significant effect, and its improving effect after IS was only observed for 'hyperarousal' behavior. Lastly, we recently developed a useful criterion, which is represented as a 'bi-directional index', for separating real PTSD rats from those exposed to IS.

Inhibitory Effects of Chlorogenic Acid on Azoxymethane-induced Colon Carcinogenesis in Male F344 Rats.

Modifying effects of chlorogenic acid (CA) on carcinogen-induced large bowel carcinogenesis was examined in rats. A total of 150 male F344 rats, 4 weeks old, were divided into 5 groups. At 6 weeks of age, groups 1-3 were given subcutaneous injections of AOM (15 mg/kg body weight) once a week for three weeks. Group 2 was given the diet mixed with CA at the dose of 250 ppm during the initiation phase (5 weeks), and group 3 was exposed to the same diet during the post-initiation phase (32 weeks). Group 4 received the diet with CA throughout the experiment. Group 5 was maintained on the basal diet alone and served as a control. At the termination of the experiment (36 weeks after the start), the incidence of colon tumors in group 2 and 3 demonstrated a tendency for decrease as compared with group 1 although this did not attain significance At this time, the multiplicity of colon tumors of group 2 was significantly smaller than in group 1. In this study, the anti-proliferating cell nuclear antigen (PCNA) indices for non-neoplastic cells of the colon mucosae in groups 2 and 3 were also smaller than in group 1. The data suggest that CA has chemopreventive potential against colon carcinogenesis in rats like that showen in a hamster model with use of methylazoxymethanol acetate.

Inhibitory Effects of Dietary Monoglucosyl-rutin on Azoxymethane-induced Colon Carcinogenesis in Rats.

The dietary effect of monoglucosyl-rutin (M-R), a flavonoid, on azoxymethane (AOM)-induced colon carcinogenesis was investigated in two experiments with 5 week old, F344 male rats. In the first experiment (5 weeks study), effects of MR on AOM (15 mg/kg body weight 3 times weekly)-induced formation of aberrant crypt foci (ACF) in five groups were assessed. In this experiment, group 3 given 500 ppm M-R with AOM had a significantly smaller number of ACF containing 4 or more aberrant crypts than group 1 with AOM alone, and groups 2 and 3 given 100 ppm or 500 ppm M-R respectively had significantly lower BrdU labeling indices in the epithelial cells of large bowel than group 1. For the second experiment, rats were divided into 8 groups. Groups 1-5 were given AOM as in the first experiment. Groups 2-5 were fed diets containing 100ppm or 500ppm M-R for 4 weeks in the initiation phase or 36 weeks in the post-initiation phase. Group 6 was given 500ppm M-R throughout the experiment, and group 7 was kept on the basal diet and served as a control. At the termination of the experiment (40 weeks after the start), groups 2-5 had significantly smaller numbers of positive cells with anti-proliferating cell nuclea antigen (PCNA) antibody than group 1. Furthermore, group 5 treated with 500ppm M-R for 36 weeks demonstrated tendencies for decrease in the incidence and multiplicity of colon tumors. These data suggest that M-R has the potential to inhibit AOM-induced colon carcinogenesis.