Optimal Number of Cycles of First-line Platinum-based Chemotherapy for Metastatic Urothelial Carcinoma.

BACKGROUND/AIM: In recent years, switch maintenance after platinum-based chemotherapy has been a standard of care. However, the appropriate number of systemic chemotherapy cycles against advanced-stage urothelial carcinoma (UC) remains unclear. This study assessed the survival outcomes of first-line platinum-based chemotherapy according to treatment cycles in patients with metastatic disease. PATIENTS AND METHODS: We retrospectively evaluated patients with metastatic bladder and upper urinary tract cancer who received platinum-based combination therapy. Overall survival (OS) was evaluated using the Kaplan-Meier method and the log-rank test. RESULTS: Of 179 patients, 47 (26.3%) were women, and 73 (40.8%) had upper urinary tract cancer. Furthermore, 47 (26.3%) who were not eligible for cisplatin received carboplatin. The median number of treatment cycles was 3 (range=1-14 cycles). The rates of progressive disease within two cycles, from two to four cycles, and from four to six cycles were 18.4%, 19.2%, and 30.6%, respectively. The median OS of patients with 2, 3, 4, 5-6, and ≥7 treatment cycles were 8.6, 14.3, 21.3, 24.4, and 26.1 months, respectively. The OS did not significantly differ between patients receiving four treatment cycles and those receiving ≥5 treatment cycles. In patients with disease control (complete or partial response or stable disease) receiving ≥4 treatment cycles, there was no significant difference in terms of OS between patients receiving four cycles and those receiving six cycles. CONCLUSION: Four cycles of first-line platinum-based chemotherapy can be effective in patients with metastatic UC.

Efficacy of Platinum-based Chemotherapy in Patients With Metastatic Urothelial Carcinoma With Variant Histology.

BACKGROUND/AIM: Variant urothelial carcinoma (VUC, defined herein as urothelial carcinoma with any histological variant) is frequently observed at an advanced stage. However, the efficacy of systemic chemotherapy against VUC in metastatic disease has rarely been reported. This study assessed the therapeutic response and survival outcomes of platinum-based chemotherapy as first-line treatment in patients with metastatic VUC. PATIENTS AND METHODS: We retrospectively analyzed consecutive patients with metastatic bladder and upper urinary tract cancer who received gemcitabine plus cisplatin (or carboplatin) at the University of Occupational and Environmental Health Hospital between November 2008 and November 2022. Progression-free survival and overall survival were evaluated using the Kaplan-Meier method and Cox proportional hazard models. RESULTS: Out of 131 patients recorded, 86 (65.6%) had pure urothelial carcinoma (PUC) and 45 (34.4%) had VUC. The most common variant element was squamous differentiation (44.4%). Compared to those with PUC, patients with VUC showed a comparable objective response rate (33.3% vs. 41.9%, p=0.451) and disease control rate (64.5% vs. 75.6%, p=0.221). They also had poorer progression-free survival (median=4.9 months vs. 7.9 months, p=0.014) and overall survival (median=10.9 months vs. 18.2 months, p=0.037) than those with PUC. On multivariate analysis, VUC was an independent predictor of progression (hazard ratio=1.79; 95% confidence interval=1.19-2.69; p=0.005) and mortality (hazard ratio=1.64; 95% confidence interval=1.08-2.48; p=0.020). CONCLUSION: Although the response of metastatic VUC to platinum-based chemotherapy was not inferior to that of PUC, VUC had progressed faster than PUC. VUC was significantly associated with a poor prognosis after platinum-based chemotherapy as first-line treatment.

Influence of submarine topography and sediment environment on microbial assemblages in a coastal lagoon in northeastern Japan.

The relationships among eutrophication, anoxia, and microbial distribution were investigated for Nagatsura-Ura Lagoon on the northeastern Pacific coast of Japan. In September 2017, the bottom environment in a small area of the inner part of the lagoon (which has a basin-shaped bottom topology) was eutrophic and anoxic, with high carbon, nitrogen, phosphate, acid-volatile sulfide, and low dissolved oxygen and oxidation-reduction potential. Dissolved oxygen levels improved during the winter. Bacillariophyta (diatoms) were the main organic component according to pigment analysis and next-generation sequencing of nucleic acids in seawater samples. Phylum Proteobacteria was dominant among the bacterial flora in the sediment but the proportions of Class Epsilon-proteobacteria and Chlorobium (a green sulfur-utilizing bacterium) were high in the inner part of the lagoon compared to other stations, and these groups were also present in winter. Apparently groups able to thrive in both anoxic and aerobic conditions were predominant in the inner part of the lagoon.

Photoinhibition and Photoprotective Responses of a Brown Marine Macroalga Acclimated to Different Light and Nutrient Regimes.

Plants and brown algae avoid photoinhibition (decline in photosystem II efficiency, Fv/Fm) caused by excess light energy and oxidative stress through several photoprotective mechanisms, such as antioxidant xanthophyll production and heat dissipation. The heat dissipation can be measured as non-photochemical quenching (NPQ) and is strongly driven by de-epoxidation of xanthophyll cycle pigments (XCP). Although NPQ is known to increase under high light acclimation and nutrient-deficient conditions, a few studies have investigated the combined effects of the conditions on both NPQ and associated xanthophyll-to-chlorophyll (Chl) a ratio. The present study investigated the photosynthetic parameters of the brown alga Sargassum fusiforme acclimated to three irradiance levels combined with three nutrient levels. Elevated irradiance decreased Fv/Fm but increased NPQ, XCP/Chl a ratio, and fucoxanthin/Chl a ratio, suggesting the photoprotective role of antioxidant fucoxanthin in brown algae. Reduced nutrient availability increased NPQ but had no effect on the other variables, including XCP/Chl a ratio and its de-epoxidation state. The results indicate that NPQ can be used as a sensitive stress marker for nutrient deficiency, but cannot be used to estimate XCP pool size and state.

Empirical estimation of marine phytoplankton assemblages in coastal and offshore areas using an in situ multi-wavelength excitation fluorometer.

Phytoplankton assemblages are essential for understanding the quality of primary production in marine ecosystems. Here, we describe the development of a methodology for monitoring marine phytoplankton assemblages using an in situ multi-wavelength excitation fluorometer (MEX). The MEX recorded the fluorescence excited with nine light-emitting diodes, temperature, and sensor depth. We prepared reference datasets comprising MEX fluorescence and plant pigment-based phytoplankton assemblages of nine chemotaxonomy groups (diatoms, dinoflagellates, cryptophytes, chlorophytes, haptophytes type 3, haptophytes type 4, prasinophytes, cyanophytes, and prochlorophytes). Conversions from the MEX fluorescence to the phytoplankton assemblages were conducted with two processes. First, target MEX fluorescence was decomposed using a linear inverse model for calculating coefficients. Second, pigment-based chemotaxonomy of the target MEX fluorescence was reconstructed using the coefficients and the chemotaxonomy assemblages of the reference data. Cross-validation analyses indicated good estimation of the proportion of diatoms, dinoflagellates, cryptophytes, cyanophytes, and prochlorophytes with MEX, and when chlorophytes, haptophytes and prasinophytes were summarized as other eukaryotes, the positive correlation was seen between proportions estimated with MEX and pigments as same as other five chemotaxonomy groups. Repeated MEX observations were conducted in the Kuroshio, the Sea of Japan, the Oyashio, and the Okhotsk Sea. The water-column integrated biomass indicated that the diatoms were an important primary producer in the Oyashio and the Okhotsk Sea, while eukaryotes were important in the Sea of Japan and prochlorophytes were important in the Kuroshio. Our method with the MEX will be a powerful tool to understand and estimate the chemotaxonomy-level assemblages and biomass in the ocean.

Analysis of environmental and microbiological changes in Onagawa Bay immediately after the tsunami of the Great East Japan Earthquake based on sediment cores.

We investigated two sediment cores to understand whether a tsunami in Onagawa Bay, Japan caused environmental changes. The value of δ13C ranged from -21.9‰ to -24.3‰ and of δ15N ranged from 5.1 to 5.9‰. We conclude that the source of the sediment in the present study area was mainly oceanic and not terrestrial. The chlorophyll concentration ranged from 1.8 to 4.0 µg/g ww, and did not vary greatly between surface and bottom layers. We inferred that all layers were deposited after the tsunami. The major phytoplankton taxa in sediments were diatoms from DNA sequencing. The presence of harmful dinoflagellates was minor. The concentrations of several heavy metals decreased slightly after the tsunami. We inferred that heavy metals in sediments were diluted by the tsunami disturbance. The land in Onagawa suffered serious damage, but disturbance of the seabed was much less evident.

Development of Ultra-Performance Liquid Chromatography with Post-Column Fluorescent Derivatization for the Rapid Detection of Saxitoxin Analogues and Analysis of Bivalve Monitoring Samples.

Saxitoxin (STX) and its analogues produced by toxic dinoflagellates accumulate in bivalves, and routine monitoring of bivalves is important to prevent cases of human poisoning. In this study, we describe a rapid detection method for the analysis of STXs using ultra-performance liquid chromatography with post-column fluorescent detection and to investigate water depths and sampling points optimal for shellfish toxin monitoring. Cultured scallops (Mizuhopecten yessoensis) and mussels (Mytilus galloprovincialis) were collected from various water depths and sampling points were used in this study. Irrespective of bivalve species, toxin concentrations in bivalves were lower at deeper water depths. The toxin concentrations of bivalves did not differ greatly when bivalves were collected from the same bay. Although the levels of contamination of bivalves with STXs can depend on various environmental and geographical factors, our findings are useful for formulating a sampling protocol for the prevention of harvesting contaminated shellfish.

Heat exposure enhances radiosensitivity by depressing DNA-PK kinase activity during double strand break repair.

PURPOSE: From the role of double strand DNA dependent protein kinase (DNA-PKcs) activity of non-homologous end joining (NHEJ) repair for DNA double strand breaks (DSBs), we aim to define possible associations between thermo-sensitisation and the enzyme activities in X-ray irradiated cells. MATERIALS AND METHODS: DNA-PKcs deficient mouse, Chinese hamster and human cultured cells were compared to the parental wild-type cells. The radiosensitivities, the number of DSBs and DNA-PKcs activities after heat-treatment were measured. RESULTS: Both DNA-PKcs deficient cells and the wild-type cells showed increased radiosensitivities after heat-treatment. The wild-type cells have two repair processes; fast repair and slow repair. In contrast, DNA-PKcs deficient cells have only the slow repair process. The fast repair component apparently disappeared by heat-treatment in the wild-type cells. In both cell types, additional heat exposure enhanced radiosensitivities. Although DNA-PKcs activity was depressed by heat, the inactivated DNA-PKcs activity recovered during an incubation at 37 °C. DSB repair efficiency was dependent on the reactivation of DNA-PKcs activity. CONCLUSION: It was suggested that NHEJ is the major process used to repair X-ray-induced DSBs and utilises DNA-PKcs activity, but homologous recombination repair provides additional secondary levels of DSB repair. The thermo-sensitisation in X-ray-irradiated cells depends on the inhibition of NHEJ repair through the depression of DNA-PKcs activities.

Microdialysis assessment of percutaneous penetration of ketoprofen after transdermal administration to hairless rats and domestic pigs.

The study was performed to evaluate the percutaneous penetration of ketoprofen after transdermal administration using a microdialysis technique in pigs, in comparison with rats. Ketoprofen release from patches was determined by analysis of the remaining drug content after application to hairless rats and pigs. Skin and knee joint penetration of ketoprofen was tested by microdialysis, and recovery was determined by retrodialysis. Residual rates in hairless rats and pigs were 68.1 ± 1.6% and 81.7 ± 4.4%, respectively, at 10h. The average recoveries of ketoprofen over 480 min in the skin and knee joint cases were 72.0 ± 3.4% and 9.8 ± 6.2% in rats and 72.3 ± 2.5% and 57.6 ± 3.1% in pigs, respectively. In rats, ketoprofen was rapidly absorbed with transdermal administration, with C(max) values of 191.7 ± 76.2 and 35.5 ± 21.7 ng/mL and AUC(0-8h) values of 918.2 ± 577.5 and 195.9 ± 137.1 ngh/mL, respectively, for the skin and knee joint. The C(max) values for the pig were 20.9 ± 18.5 and 3.7 ± 3.0 ng/mL, with AUC(0-8h) values of 73.1 ± 69.2 and 16.1 ± 16.1 ngh/mL. Ketoprofen concentrations within skin and knee joint of non-application sites in rats and pigs were less than 0.8 ng/mL. Transdermal administration of ketoprofen significantly reduced prostaglandin E2 levels in the skin of the application site and showed a tendency for inhibition in the knee joint. We thus demonstrated that topical patches containing ketoprofen can deliver the drug through the skin and knee joint of pigs and rats via direct diffusion, and microdialysis data with the pig may be useful for the prediction of human tissue penetration of drugs with transdermal administration.

Camptothecin disrupts androgen receptor signaling and suppresses prostate cancer cell growth.

The androgen receptor (AR) is the main therapeutic target for treatment of metastatic prostate cancers. The present study demonstrates that the topoisomerase I inhibitor camptothecin selectively inhibits androgen-responsive growth of prostate cancer cells. Camptothecin strikingly inhibited mutated and wild-type AR protein expression in LNCaP and PC-3/AR cells. This inhibition coincided with decreased androgen-mediated AR phosphorylation at Ser(81) and reduced androgen-mediated AR transcriptional activity in a dose-dependent manner. Additionally, camptothecin disrupted the association between AR and heat shock protein 90 and impeded binding of the synthetic androgen [3H]R1881 to AR in LNCaP cells. Camptothecin also blocked androgen-induced AR nuclear translocation, leading to downregulation of the AR target gene PSA. In addition to decreasing the intracellular and secreted prostate-specific antigen (PSA) levels, camptothecin markedly inhibited androgen-stimulated PSA promoter activity. Collectively, our data reveal that camptothecin not only serves as a traditional genotoxic agent but, by virtue of its ability to target and disrupt AR, may also be a novel candidate for the treatment of prostate cancer.

Variations in sensitivity to ionizing radiation in relation to p53 mutation point.

Mutation of p53 is the most common genetic alteration observed in human tumours and is reported to lead to variations in cell radiosensitivity. However, the relationship between the mutation point and the degree of radiosensitivity is unclear. Saos-2 cells with different mutations of p53 were prepared and examined for radiosensitivity. Cells with p53 mutations at codons 175, 244, 245, 273 and 282 were radioresistant, whereas those with mutations at codons 123, 195, 238 and 242 were radiosensitive. Mutations at codons 130, 143, 157, 168, 277, 280 and 286 resulted in medium radiosensitivity. Thus the sensitivity of Saos-2 cells to ionizing radiation varies with the mutation point of the p53 gene.

The greater lethality of UVB radiation to cultured human cells is associated with the specific activation of a DNA damage-independent signaling pathway.

UV radiation causes cell death through the activation of various intracellular signaling molecules in both DNA damage-dependent and -independent manners. The ability of middle-wavelength UV (UVB) radiation to form DNA photoproducts is less than that of short-wavelength UV (UVC) radiation; however, the differences between UVB and UVC radiation in the extent of DNA damage-independent signaling and its contribution to cell death have not been well characterized. When cells were irradiated with UVB or UVC radiation at doses that generated equivalent amounts of DNA photoproducts, UVB radiation induced more clonogenic cell death, apoptotic cells, mitochondrial cytochrome C release, and intracellular oxidative stress. Among the signaling molecules examined, levels of p53 phosphorylated at Ser-392 and p38 were higher in UVB-irradiated cells than in UVC-irradiated cells. Both phosphorylations were reduced by treating cells with an antioxidant. Furthermore, an inhibitor of p38 also blocked the phosphorylation of p53 at Ser-392. These results suggest that UVB radiation activates the p38 pathway through the generation of oxidative stress, which merges with the DNA p53 pathway by phosphorylation of p53 at ser392. This greater contribution of the DNA damage-independent pathway in UVB-irradiated cells may explain the greater lethality of UVB radiation.

Evaluation of fat-free mass by whole-body counter in Japanese healthy young adults.

Whole-body counters (WBCs) are special instruments for measuring internal irradiation doses and are usually housed within or around nuclear facilities in the event of unexpected radiation emergencies. As a substantial proportion of total body potassium (TBK) is found in fat-free mass (FFM), FFM volume can be predicted from WBC-measured (40)K. We screened TBK in Japanese healthy young adults using a WBC and found strong linear correlations between TBK and lean body mass (LBM) and body mass index (r = 0.97, P < 0.01 and r = 0.47, P < 0.01, respectively). Multiple linear regression analysis, following adjustments for sex, indicates that only LBM has a significant correlation with TBK (P < 0.01). These results strongly support the feasibility of using WBCs for estimating FFM.

Sucralfate protects intestinal epithelial cells from radiation-induced apoptosis in rats.

Radiotherapy for malignant pelvic disease is often followed by acute radiation colitis (ARC). It has been reported that sucralfate treatment has a protective effect against ARC, though the mechanisms of action are unknown. The effects of sucralfate on X-ray radiation-induced apoptosis was studied at 4 Gy in the colonic crypt cells of rats. Sucralfate enemas given prior to radiation resulted in the following: (1) reduction in number of apoptotic colonic crypt cells; (2) reduction in number of caspase-3 positive cells; (3) decreases in p53 accumulation and p21 expression; (4) decreases of Bax/Bcl-2 ratio. The protective effects of sucralfate against ARC may be partially due to the suppression of radiation-induced apoptosis by way of p53 in the colon and the protection of the colonic epithelial stem cell region.

Phototoxicity study of a ketoprofen poultice in guinea pigs.

Ketoprofen has been reported to have such side effects as photosensitive dermatitis in humans (The Ministry of Health, Labour and Welfare, 2001). In the present study, as part of a safety evaluation of Miltax, an application drug containing ketoprofen, phototoxicity of Miltax was examined in guinea pigs. In the present skin phototoxicity study, Miltax was applied for 12 hr. Ultraviolet (UV) rays were irradiated to examine whether or not Miltax elicited skin reaction. Two kinds of UV-A plus UV-B dual irradiation and UV-A single irradiation were used for the elicitation. With UV-A plus UV-B dual irradiation on the Miltax application site, no skin reaction was observed at UV irradiation side in any animals, in contrast to the case of the positive control article, 8-methoxypsoralen (8-MOP). Similar results were obtained with UV-A single irradiation. From these results, Miltax that contained ketoprofen did not show any skin phototoxicity in the guinea pig.

Measurement of the whole-body 137Cs in residents around the Chernobyl nuclear power plant.

To understand the current situation of internal radiation exposure in the population around the Chernobyl Nuclear Power Plant (CNPP), we examined the 137Cs body burden in six residents of Belarus, Ukraine and Russia in 2002 and 2004 using the whole-body counter (WBC) at Nagasaki University (Japan). The data were compared with those of our previous study performed in 1993-1994 using the same method. In 2002 and 2004, peaks of 137Cs were detected in two residents from Gomel, which was heavily contaminated by the CNPP accident, one from Minsk (Belarus) and one from Kiev (Ukraine), but another resident from Minsk showed no 137Cs peaks. The results of the present study suggests that residents around the CNPP are still exposed to chronic 137Cs internal irradiation, probably due to the daily consumption of contaminated domestic foods, but the risk of any disease by the irradiation is quite low. Long-term follow-up of WBC around the CNPP is useful and may contribute to radiation safety regulation together with a reduction of unnecessary radiophobia for the residents.

Hypergravity induces phosphorylation of p53 at serine 15, but not an expression of p53-downstream genes.

We investigated the p53 signaling pathway induced by hypergravity in the human glioblastoma cell line A172. Hypergravity (20 x g) induced the accumulation of p53 and the phosphorylation of p53 at Ser-15. The phosphorylation of p53 with hypergravity was not inhibited by wortmannin, the PI3-kinase inhibitor. This indicated that the p53 signal pathway induced by hypergravity is different from other p53 signal pathways, such as that of the DNA damage signal. Hypergravity did not induce an expression of the genes Waf-1 or Bax, located downstream from p53. We also examined the expression of genes with hypergravity by using a DNA microarray containing oligo DNA from 30,000 human genes. Hypergravity (20 x g, 6 h) did induce the expression of some genes concerned with the cell signaling pathway and cytoskeleton of the cell, but not any of the p53-downstream genes. DNA microarray revealed the induction of many genes to enable the cells to adapt to the hypergravity environment.

Concentrations of polychlorinated dibenzo-p-dioxins, dibenzofurans, non-ortho polychlorinated biphenyls, and mono-ortho polychlorinated biphenyls in Japanese flounder, with reference to the relationship between body length and concentration.

The concentrations of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), non-ortho polychlorinated biphenyls (non-ortho PCBs), and mono-ortho polychlorinated biphenyls (mono-ortho PCBs) in Japanese flounder (Paralichthys olivaceus, body length 10.4-36.6 cm) collected from Sendai Bay, Japan, were determined using high-resolution gas chromatography-mass spectrometry. The relationship between the concentrations of these compounds (dioxins) and the body length of the Japanese flounder was examined. The total PCDD and total PCDF concentrations did not correlate with body length (both r(2) < 0.1, both p > 0.05), whereas the total non-ortho PCB and total mono-ortho PCB (coplanar polychlorinated biphenyls, Co-PCBs) concentrations were significantly correlated (r(2)= 0.8, p < 0.05 and r(2)= 0.63, p < 0.05, respectively). The bioaccumulation properties of PCDD/Fs in Japanese flounder differed from those of Co-PCBs. Toxicity equivalency quotient (TEQ) values derived from the Co-PCBs made up 46.3%-63.7% of the total TEQ value for all the dioxins. Although the concentrations of non-ortho PCBs were lower than those of mono-ortho PCBs, the TEQ value for non-ortho PCBs was higher than that for mono-ortho PCBs. The TEQ value for non-ortho PCBs increased more with increasing body length than did the values for PCDDs, PCDFs, and mono-ortho PCBs. These results show that from the standpoint of risk management, non-ortho Co-PCBs are the most important of the dioxins in Japanese flounder.

Multicentric adenomatoid tumors involving uterus, ovary, and appendix.

A case of large, and multicentric adenomatoid tumors involving the uterus, right ovary and appendix is described. The uterine tumor was exceptionally large, about 15 cm in maximal diameter, and located in the right fundal region of the uterus with a short, thick pedicle as in a subserous pedunculated uterine fibroid.

Sources of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and coplanar polychlorinated biphenyls (Co-PCBs), and their bioaccumulation through the marine food web in Sendai Bay, Japan.

The concentrations of PCDD/F and Co-PCB congeners in seawater, sediment, Pacific oyster, Japanese anchovy, marbled sole, and Japanese flounder samples from Sendai Bay, Japan, were analyzed. The compositions of total PCDD/F and total Co-PCB concentrations in sediment and Pacific oyster reflected that in seawater--the compositions of total PCDD, PCDF, and Co-PCB concentrations were approximately 60, 10, and 30%, respectively. The compositions in Japanese anchovy, marbled sole, and Japanese flounder were different from those in seawater and sediment-the ratio of total Co-PCB concentration to total PCDD/F plus Co-PCB concentrations in Japanese anchovy, marbled sole, and Japanese flounder was above 90%. Tetrachlorinated PCDDs (T4CDDs), such as 1,3,6,8- and 1,3,7,9-T4CDD, were the predominant congeners in seawater and sediment; the total T4CDD concentrations in seawater and sediment were approximately 46 and 48% of the total PCDD concentration. Furthermore, shipments of the herbicide 1,3,5-trichloro-2-(4-nitrophenoxy)benzene to Miyagi Prefecture, the so-called granary of Japan, were the highest in Japan over the last 12 years. The major sources of PCDD/Fs and Co-PCBs in Sendai Bay, which is in Miyagi Prefecture, are impurities in chlorinated herbicides. The order of concentration of PCDD/Fs was Pacific oyster > Japanese anchovy = marbled sole > Japanese flounder; the concentration in Japanese flounder, which is a higher-trophic-level consumer in the marine food web, was lower than that in shellfish (Pacific oyster) and Japanese anchovy, which are lower-trophic-level consumers. The order of concentration of Co-PCBs was Pacific oyster < Japanese anchovy = marbled sole < Japanese flounder; the concentrations in the higher-trophic-level consumers were higher than the concentrations in the lower-trophic-level consumers. Different PCDD/F congeners tended to bioaccumulate in different organisms. On the other hand, all species of Co-PCB congener tended to bioaccumulate in all organisms.