A preliminary report on retrospective dose assessment by FISH translocation assay in FDNPP Nuclear Emergency Worker Study (NEWS).

In Japan, a national project of longitudinal health care and epidemiological research (NEWS) was developed in 2014 to analyse the effects of radiation on human health for workers who responded to the Fukushima Dai-ichi nuclear emergency in 2011. In 2018, peripheral blood for chromosome translocation analysis was collected from 62 workers. Retrospective dose assessment was performed with fluorescence in situ hybridisation translocation (FISH-Tr) assay. The range of estimated doses by FISH-Tr assay was 0-635 mGy, in which 22 workers had estimated doses of more than 189 mGy. Biological dose estimates were five times higher in workers with physically measured total exposure recordings above 70 mGy. It is likely that smoking and medical exposure caused the discrepancy between estimated biological and physical total exposure doses. Thus, there is a possibility that retrospective biodosimetry assessment might over-estimate occupational exposures to workers exposed to chronic radiation during nuclear emergency work.

ΔNp63α transcriptionally represses p53 target genes involved in the radiation-induced DNA damage response : ΔNp63α may cause genomic instability in epithelial stem cells.

BACKGROUND: The DNA damage response (DDR) is a mechanism that protects cells against radiation-induced oxidative DNA damage by causing cell cycle arrest and apoptosis. TP63 is a member of the tumour suppressor TP53 gene family, and ΔNp63α, a TP63 splicing variant, is constitutively expressed in the stem cell-containing basal layer of stratified epithelial tissues, including the mammary gland, where it plays a critical role in stemness and tissue development. ΔNp63α has been reported to transcriptionally inhibit the tumour suppression protein p53. This p53-repressive activity may cause genomic instability in epithelial stem cells exposed to radiation. In this study, we analysed the inhibitory effect of ΔNp63α on radiation-induced DDR. METHODS: To elucidate the role of the p53-repressive effect of ΔNp63α in radiation response, we performed a p63-siRNA knockdown experiment using human mammary epithelial cells (HMECs) expressing ΔNp63α and then performed ectopic and entopic expression experiments using human induced pluripotent stem cells (hiPSCs). After irradiation, the expression of DDR-related genes and proteins in ΔNp63α-expressing and control cells was analysed by RT-qPCR, Western blotting, and flow cytometry. RESULTS: The mRNA/protein expression levels of BAX and p21 were significantly increased in p63-siRNA-treated HMECs (sip63) after X-ray irradiation (4 Gy, 0.7 Gy/min) but not in scramble-siRNA treated HMECs (scr). Transcriptomic analysis showed decreased RNA expression of cell cycle-related genes and increased expression of programmed cell death-related genes in sip63 cells compared to scr cells. Furthermore, flow cytometric analysis revealed an increase in apoptotic cells and a decrease in 5-ethynyl-2´-deoxyuridine uptake in sip63 cells compared to scr cells. On the other hand, both the ectopic and entopic expression of ΔNp63α in apoptosis-sensitive hiPSCs reduced the expression levels of BAX after irradiation and significantly decreased the number of apoptotic cells induced by radiation. CONCLUSION: Taken together, these results indicate that ΔNp63α represses p53-related radiation-induced DDR, thereby potentially causing genomic instability in epithelial stem cells.

DOSE-RATE EFFECT OF RADIATION ON RAT MAMMARY CARCINOGENESIS AND AN EMERGING ROLE FOR STEM CELL BIOLOGY.

The uncertain cancer risk of protracted radiation exposure at low dose rates is an important issue in radiological protection. Tissue stem/progenitor cells are a supposed origin of cancer and may contribute to the dose-rate effect on carcinogenesis. The authors have shown that female rats subjected to continuous whole body γ irradiation as juveniles or young adults have a notably reduced incidence of mammary cancer as compared with those irradiated acutely. Experiments using the mammosphere formation assay suggested the presence of radioresistant progenitor cells. Cell sorting indicated that basal progenitor cells in rat mammary gland were more resistant than luminal progenitors to killing by acute radiation, especially at high doses. Thus, the evidence indicates a cell-type-dependent inactivation of mammary cells that manifests only at high acute doses, implying a link to the observed dose-rate effect on carcinogenesis.

Comparison of the Mental Burden on Nursing Care Providers With and Without Mat-Type Sleep State Sensors at a Nursing Home in Tokyo, Japan: Quasi-Experimental Study.

BACKGROUND: Increasing need for nursing care has led to the increased burden on formal caregivers, with those in nursing homes having to deal with exhausting labor. Although research activities on the use of internet of things devices to support nursing care for older adults exist, there is limited evidence on the effectiveness of these interventions among formal caregivers in nursing homes. OBJECTIVE: This study aims to investigate whether mat-type sleep state sensors for supporting nursing care can reduce the mental burden of formal caregivers in a nursing home. METHODS: This was a quasi-experimental study at a nursing home in Tokyo, Japan. The study participants were formal caregivers who cared for residents in private rooms on the fourth and fifth floors of the nursing home. In the intervention group, formal caregivers took care of residents who used sleep state sensors on the fourth floor of the nursing home. The sleep state sensors were mat types and designed to detect body motion such as the frequency of toss and turning and to measure heartbeat and respiration. One sensor was placed on a bed in a private room. When body motion is detected, the information is instantly displayed on a monitor at a staff station. In addition, the mental condition of the formal caregivers was measured using a validated self-reported outcome measure-the Profile of Mood States (POMS), Short-Form, 2nd edition. Formal caregivers in both groups received the POMS at baseline, midpoint (week 4), and endpoint (week 8) to identify changes in these domains. The primary outcome was the difference in total mood disturbance (TMD) of the POMS at baseline and week 8. RESULTS: Of the 22 eligible formal caregivers, 12 (intervention group) utilized sleep state sensors for 8 weeks. The remaining 10 formal caregivers (control group) provided nursing care as usual. As for the primary outcome of the difference between TMD at baseline and week 8, TMD in the intervention group improved by -3.67 versus 4.70 in the control group, resulting in a mean difference of -8.37 (95% CI -32.02 to 15.29; P=.48) in favor of the intervention. CONCLUSIONS: The present 8-week study showed that sleep state sensing for elderly residents might not be associated with reduced mental burdens on formal caregivers in nursing homes.

Clinical impact of patient-centered care for hemodialysis patients using routine assessment of symptom burden.

The study aimed to verify the impact of our clinical strategy, which emphasizes patient-centered care, based on patient-reported outcome measures (PROMs) results in hemodialysis patients. We developed our original PROM (comprising 20 items) to assess patients' symptom burden. To confirm the validity of our clinical pattern, we performed various analyses using PROM data. We retrospectively enrolled 383 individuals (mean age 66.3 years; 252 men), collected their PROM data in December 2013, and followed them up for 3 years. We noted a lower mortality rate and a lower prevalence of itching in our facilities than in previous surveys and reports in Japan. Furthermore, we observed that the severity of symptom burden affected medium-term prognosis. This is the first study to report the results of patient-centered medical practice utilizing PROMs in dialysis care. Careful attention should be paid to patients' symptom burden, as performed in objective data management.

Chromosomal translocation t(11;14) and p53 deletion induced by the CRISPR/Cas9 system in normal B cell-derived iPS cells.

Multiple myeloma (MM) cells are derived from mature B cells based on immunoglobulin heavy chain (IgH) gene analysis. The onset of MM is often caused by a reciprocal chromosomal translocation (cTr) between chr 14 with IgH and chr 11 with CCND1. We propose that mature B cells gain potential to transform by reprograming, and then chromosomal aberrations cause the development of abnormal B cells as a myeloma-initiating cell during B cell redifferentiation. To study myeloma-initiating cells, we have already established normal B cell-derived induced pluripotent stem cells (BiPSCs). Here we established two BiPSCs with reciprocal cTr t(11;14) using the CRISPR/Cas9 system; the cleavage site were located in the IgH Eµ region of either the VDJ rearranged allele or non-rearranged allele of IgH and the 5'-upsteam region of the CCND1 (two types of BiPSC13 with t(11;14) and MIB2-6 with t(11;14)). Furthermore, p53 was deleted using the CRISPR/Cas9 system in BiPSC13 with t(11;14). These BiPSCs differentiated into hematopoietic progenitor cells (HPCs). However, unlike cord blood, those HPCs did not differentiated into B lymphocytes by co-culture with BM stromal cell. Therefore, further ingenuity is required to differentiate those BiPSCs-derived HPCs into B lymphocytes.

Flow Cytometry Definition of Rat Mammary Epithelial Cell Populations and Their Distinct Radiation Responses.

Breast tissue is very susceptible to radiation-induced carcinogenesis, and mammary stem/progenitor cells are potentially important targets of this. The mammary epithelium is maintained as two mostly independent lineages of luminal and basal cells. To elucidate their immediate radiation responses, we analyzed the mammary glands of female Sprague-Dawley rats, a radiation carcinogenesis model, using colony formation, flow cytometry and immunofluorescence. The results revealed that flow cytometry successfully fractionates rat mammary cells into CD49fhi CD24lo basal, CD49fmed CD24hi luminal progenitor, and CD49flo CD24hi mature luminal populations, resembling human breast, rather than mouse tissues. The colony-forming ability of the basal cells was more radiosensitive than the luminal progenitor cells. Flow cytometry and immunofluorescence showed more efficient cell cycle arrest, γ-H2AX responses, and apoptosis in the irradiated luminal progenitor cells, than in the basal cells. These results provide important insights into the early phase of radiation-induced breast cancer.

Development of a gateway for interoperability in community-based care: An empirical study.

BACKGROUND: Information and communications technology has attracted attention as a useful way of sharing care records in community-based care. Such information sharing systems, however, imposed the burden of inputting the same records into different information systems due to a lack of interoperability of the systems. OBJECTIVES: The purpose of this study was to develop a gateway that links information systems and to investigate the functionality and usability of the gateway through an empirical study. METHODS: We developed a gateway with healthcare and welfare professionals in Kashiwa city, Japan. The gateway system consisted of two sub-systems: a data exchange sub-system and a common sub-system. Regarding the security, we used the transport layer security 1.2 and a public key infrastructure. For document formats, we utilized the health level seven international, extensible markup language, and portable document format. In addition, we performed an empirical study with 11 scenarios of four simulated patients and a questionnaire survey to the professionals. RESULTS: Professionals of eight occupations participated the empirical study and verified the gateway to link information systems of six vendors. For a questionnaire survey, 32 professionals out of 40 reported that the gateway would eliminate the burden of inputting the same records into different information systems.

Isolation of chicken taste buds for real-time Ca2+ imaging.

We isolated chicken taste buds and used a real-time Ca2+ imaging technique to investigate the functions of the taste cells. With RT-PCR, we found that isolated chicken taste bud-like cell subsets express chicken gustducin messenger RNA. Immunocytochemical techniques revealed that the cell subsets were also immunopositive for chicken gustducin. These results provided strong evidence that the isolated cell subsets contain chicken taste buds. The isolated cell subsets were spindle-shaped and approximately 61-75 µm wide and 88-98 µm long, and these characteristics are similar to those of sectional chicken taste buds. Using Ca2+ imaging, we observed the buds' response to 2 mmol/L quinine hydrochloride (a bitter substance) and their response to a mixture of 25 mmol/L L-glutamic acid monopotassium salt monohydrate and 1 mmol/L inosine 5'-monophosphate disodium salt, umami substances. The present study is the first morphological demonstration of isolated chicken taste buds, and our results indicate that the isolated taste buds were intact and functional approaches for examining the taste senses of the chicken using Ca2+ imaging can be informative.

Chronic hemodialysis patients without marked elevation of intact parathyroid hormone are also good candidates for early intervention with cinacalcet.

Management of calcium (Ca) and phosphorus (P) metabolism is crucial in chronic hemodialysis (HD) patients. Cinacalcet is usually used for chronic kidney disease-mineral and bone disorders (CKD-MBD) patients with elevated intact parathyroid hormone (iPTH) levels. However, a certain number of CKD-MBD patients have normal iPTH levels and are not subjected to cinacalcet therapy. Here, we evaluated the efficacy of a new treatment algorithm of early initiation of cinacalcet therapy in this subgroup of patients, mainly for correcting Ca and P metabolism. Seventy-one HD patients, including 44 patients without marked elevation of iPTH (102 < iPTH ≤ 300 pg/mL), who received cinacalcet therapy, were enrolled in this study. Serum parameters relating to CKD-MBD patient metabolism, doses of phosphate binders, and type of vitamin D sterols were compared between pre- and post-cinacalcet administration retrospectively. Sixty-four of 71 patients did not require discontinuation of cinacalcet. In these 64 patients, serum Ca (P = 0.0003), P (P = 0.0153), and iPTH (P < 0.0001) levels were significantly reduced after cinacalcet administration, even in those without marked elevation of iPTH (Ca; P < 0.0001, P; P = 0.0422, and iPTH; P = 0.0018). The proportion of patients who received vitamin D sterols was unchanged (P = 0.5930) but the proportion of patients who received maxacalcitol was significantly reduced after cinacalcet administration (P = 0.0108). The new treatment algorithm of early initiation of cinacalcet is considered to be well tolerated and effective for controlling hypercalcemia, and/or hyperphosphatemia and/or increased iPTH of CKD-MBD patients.

Gustducin is expressed in the taste buds of the chicken.

We investigated the expression of gustducin in chicken taste buds using molecular biological, biochemical and immunohistochemical techniques. Expression of a gustducin-like sequence was detected by RT-PCR in the tissues containing taste buds, and corresponded to the predicted gustducin gene in the chicken. Expression of this sequence was not detected in the brain, heart, liver, pancreas, intestine, kidney and testis of the chicken. The expressed sequence had a high specificity for oral tissues that contained taste buds. These results suggest that the detected sequence was the chicken gustducin gene. Next, we generated a polyclonal antiserum against the chicken gustducin protein to observe its localization in the oral tissues. The results revealed that the chicken gustducin was specifically expressed in the taste buds. It is suggested that the chicken has a gustatory system mediated by gustducin, and chicken gustducin is a reliable marker for taste buds or taste cells. This is the first molecular biological, biochemical and immunohistochemical demonstration of the presence of gustducin in the chicken.

The number of taste buds is related to bitter taste sensitivity in layer and broiler chickens.

The relationship between taste sensitivity and the number of taste buds using a bitter tastant, quinine hydrochloride, was investigated in White Leghorn, Rhode Island Red, and broiler chickens. The White Leghorn and Rhode Island Red strains were able to perceive 2.0 mmol/L quinine hydrochloride, but the taste sensitivity of Rhode Island Red chickens was higher than that of White Leghorn chickens. Broiler chickens perceived 0.5 mmol/L quinine hydrochloride. The number of taste buds in the White Leghorn strain was the lowest, then the Rhode Island Red strain, with the number of taste buds highest in the broiler chickens. The number of taste buds was well correlated with bitter taste sensitivity. Therefore, we suggest that the number of taste buds is a vital factor in the perception of bitter taste and may be useful in selecting appropriate feeds for chickens.

Comprehensive survey of p94/calpain 3 substrates by comparative proteomics--possible regulation of protein synthesis by p94.

Calpain represents a family of Ca(2+)-dependent cytosolic cysteine proteases found in almost all eukaryotes and some bacteria, and is involved in a variety of biological phenomena, including brain function. Several substrates of calpain are aggressively proteolyzed under pathological conditions, e.g., in neurodegenerating processes, fodrin is proteolyzed by calpain. Because very small amounts of substrate are proteolyzed by calpain under normal biological conditions, the molecular identities of calpain substrates are largely unknown. In this study, an extensive survey of the substrates of p94/calpain 3 in COS7 cells was executed using iTRAQ(TM) labeling and 2-D LC-MALDI analysis. p94 was used because: (i) several p94 splicing variants are expressed in brain tissue even though p94 itself is a skeletal-muscle-specific calpain, and (ii) it exhibits Ca(2+)-independent activity in COS cells, which makes it useful for evaluating the effects of p94 protease activity on proteins without perturbing the cells. Our approach revealed several novel protein substrates for p94, including the substrates of conventional calpains, components of the protein synthesis system, and enzymes of the glycolytic pathway. The results demonstrate the usefulness and sensitivity of this approach for mining calpain substrates. A combination of this method with other analytical methods would contribute to elucidation of the biological relevance of the calpain family.

Structure of bovine fungiform taste buds and their immunoreactivity for gustducin.

The taste buds of bovine fungiform papillae were studied by light and electron microscopy using both histological and immunohistochemical methods. The taste buds existed in the epithelium of the apical region of the papillae. By electron microscopy, two types of taste cells, namely type I and type II cells, could be classified according to the presence of dense-cored vesicles, the cytoplasmic density and the cell shape. Type I cells were thin, had an electron-dense cytoplasm containing dense-cored vesicles, and possessed long thick apical processes in the taste pore. Type II cells were thick, had an electron-lucent cytoplasm containing many electron-lucent vesicles, rather than dense-cored vesicles, and possessed microvilli in the taste pore. Immunohistochemical staining with an antiserum against gustducin was investigated by both light and electron microscopy using the avidin-biotin complex (ABC) method. Some, but not all, of the type II cells exhibited gustducin immunoreactivity, whereas none of the type I cells showed any immunoreactivity.

A case of exercise-induced acute renal failure in a patient with idiopathic renal hypouricemia developed during antihypertensive therapy with losartan and trichlormethiazide.

Exercise-induced acute renal failure (ARF) developed in a 45-year-old man during antihypertensive therapy with losartan and trichlormethiazide. The antihypertensive therapy was stopped and marked hypouricemia became apparent during improvement of his renal function. The daily urinary excretion of uric acid was normal and an increased fractional excretion of uric acid was observed. Renal biopsy revealed that the kidney was recovering from acute tubular necrosis with interstitial fibrosis. Based on the results of pyrazinamide and benzbromarone tests, we classified this case as one of presecretory reabsorption defect of uric acid. Antihypertesive therapy with benidipine and candesartan was initiated, and the patient has not had any ARF episodes since. Because idiopathic renal hypouricemia can be associated with exercise-induced ARF and chronic renal dysfunction, careful antihypertensive therapy and follow-up evaluation of renal function might be necessary for hypertensive patients with idiopathic renal hypouricemia.