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Nishioka S.,Kagawa University | Nakano D.,Kagawa University | Kitada K.,Kagawa University | Sofue T.,Kagawa University | And 6 more authors.
Kidney International | Year: 2014

The cyclin-dependent kinase inhibitor p21 plays important roles in chronic renal disorders; however, its roles in response to acute renal stress are unclear. Here we evaluated p21 in acute kidney injury and ischemic preconditioning using wild-type and p21 knockout mice that underwent renal ischemia followed by reperfusion. The decline in renal function and histological changes were worse in the knockout than in wild-type mice. Ischemia/reperfusion increased p21 expression in the kidney of wild-type mice compared with sham surgery, suggesting p21 may confer tolerance to ischemia/reperfusion injury. We next tested whether p21 is associated with the protective effect of ischemic preconditioning, an established method to reduce ischemia/reperfusion injury. Ischemic preconditioning attenuated ischemia/reperfusion injury in wild-type but not p21-knockout mice. This preconditioning decreased the number of proliferating tubular cells before but increased them at 24 h after ischemia/reperfusion in the kidneys of wild-type mice. In p21-knockout mice, ischemic preconditioning did not change the number of proliferating cells before but decreased them after ischemia/reperfusion. Ischemic preconditioning increased renal p21 expression and the number of cells in the G1 phase of the cell cycle before ischemia/reperfusion compared with sham surgery. Thus, renal p21 is essential for the beneficial effects of renal ischemic preconditioning. Transient cell cycle arrest induced by ischemic preconditioning by a p21-dependent pathway seems to be important for subsequent tubular cell proliferation after ischemia/reperfusion. © 2013 International Society of Nephrology.

Su J.,Ehime University | Hato-Yamada N.,Ehime Prefectural University of Health Sciences | Araki H.,Ehime University | Yoshimura H.,Research Institute for Alternative Medicine
Journal of Pharmacological Sciences | Year: 2013

The forced swimming test (FST) in mice is widely used to predict the antidepressant activity of a drug, but information describing the immobility of female mice is limited. We investigated whether a prior swimming experience affects the immobility duration in a second FST in female mice and whether the test-retest paradigm is a valid screening tool for antidepressants. Female ICR mice were exposed to the FST using two experimental paradigms: a single FST and a double FST in which mice had experienced FST once 24 h prior to the second trail. The initial FST experience reliably prolonged immobility duration in the second FST. The antidepressants imipramine and paroxetine significantly reduced immobility duration in the single FST, but not in the double FST. Scopolamine and the sigma-1 (σ1) antagonist NE-100 administered before the second trial significantly prevented the prolongation of immobility. Neither a 5-HT1A nor a 5-HT2A receptor agonist affected immobility duration. We suggest that the test-retest paradigm in female mice is not adequate for predicting antidepressant-like activity of a drug; the prolongation of immobility in the double FST is modulated through acetylcholine and σ1 receptors. © The Japanese Pharmacological Society.

Norimatsu Y.,Ehime Prefectural University of Health Sciences | Yanoh K.,Suzuka General Hospital | Kobayashi T.K.,Osaka University
Acta Cytologica | Year: 2013

Objective: Liquid-based preparation (LBP) of the endometrial lesions is an important diagnostic tool for a variety of endometrial abnormalities because of its simplicity and high quali-quantitative diagnostic yield. We aimed to investigate the LBP method for endometrial cytology to evaluate both benign and abnormal endometrial lesions. Study Design: LBP is a semiautomated methodology that has recently become widely available and has gained popularity as a method of collecting and processing both gynecologic and nongynecologic cellular specimens. Results: Some peculiar endometrial cytoarchitectural features were described using LBPs. These were advantageous to screen as compared to conventional slides due to a smaller screening area and an excellent quality of cell preparations. Conclusions: LBP is a useful tool in the cellular diagnosis and follow-up of endometrial abnormalities, which remains complementary to the emerging molecular diagnostic cytopathology. The study of LBPs from endometrial cytology could be challenging since it is affected by numerous look-alikes and diagnostic pitfalls. This review discusses these various entities and takes into consideration the ancillary techniques that may be useful in the diagnostic procedure. © 2013 S. Karger AG, Basel.

Takata T.,Ehime Prefectural University of Health Sciences | Takata T.,Okayama University of Science | Morimoto C.,Matsuyama Shinonome College
Journal of Medicinal Food | Year: 2014

The decrease in the bone mass associated with osteoporosis caused by ovariectomy, aging, and other conditions is accompanied by an increase in bone marrow adipose tissue. The balance between osteoblasts and adipocytes is influenced by a reciprocal relationship. The development of modalities to promote local/systemic bone formation by inhibiting bone marrow adipose tissue is important in the treatment of fractures or metabolic bone diseases such as osteoporosis. In this study, we examined whether raspberry ketone [4-(4-hydroxyphenyl)butan-2-one; RK], which is one of the major aromatic compounds of red raspberry and exhibits anti-obesity action, could promote osteoblast differentiation in C3H10T1/2 stem cells. Confluent C3H10T1/2 stem cells were treated for 6 days with 10-100 μg/mL of RK in culture medium containing 10 nM all-trans-retinoic acid (ATRA) or 300 ng/mL recombinant human bone morphogenetic protein (rhBMP)-2 protein as an osteoblast-differentiating agent. RK in the presence of ATRA increased alkaline phosphatase (ALP) activity in a dose-dependent manner. RK in the presence of rhBMP-2 also increased ALP activity. RK in the presence of ATRA also increased the levels of mRNAs of osteocalcin, α1(I) collagen, and TGF-βs (TGF-β1, TGF-β2, and TGF-β3) compared with ATRA only. RK promoted the differentiation of C3H10T1/2 stem cells into osteoblasts. However, RK did not affect the inhibition of early-stage adipocyte differentiation. Our results suggest that RK enhances the differentiation of C3H10T1/2 stem cells into osteoblasts, and it may promote bone formation by an action unrelated to adipocyte differentiation. © 2014, Mary Ann Liebert, Inc.

Kitao T.,Ehime Prefectural University of Health Sciences | Kitao T.,Okayama University | Ishimaru M.,Ehime Prefectural Central Hospital | Nishihara S.,Ehime Prefectural Central Hospital
Journal of Infection and Chemotherapy | Year: 2010

The detection of biofilm-producing (ica AB) and methicillin resistance genes (mec A) was investigated in 70 blood culture isolates of Staphylococcus epidermidis and in 66 and 51 isolates from human hands and the vestibules of the nose, respectively, of 77 healthy subjects who gave consent. Of the 70 strains isolated from blood culture testing, both ica AB and mec A were detected in 36 (51.4%), and neither was detected in 4 (5.7%). The mec A gene only was detected in 30 (42.9%), but no isolate from blood culture testing possessed the ica AB gene alone. In contrast, of the 66 isolates from healthy hands, only one isolate (1.5%) possessed both genes, whereas neither was detected in 56 (84.8%), but the mec A gene was detected in 9 (13.6%). Of the 51 isolates from the nasal vestibules, both genes were detected in 12 (23.5%), and neither in 15 (29.4%). Moreover, the mec A gene was detected in 17 (33.3%). Thus, S. epidermidis strains that normally inhabit the nasal vestibule were found to carry the ica AB and mec A genes more frequently than those that inhabit the fingers. The ica AB and mec A genes were detected in S. epidermidis isolated in blood culture tests from patients diagnosed with sepsis associated with catheter-related bloodstream infection (CR-BSI). Both genes were detected in 7 (70.0%) of 10 isolates, and the mec A gene alone was detected in 3 (30.0%). In fact, we could not detect any strain carrying only the ica AB gene from S. epidermidis, an infecting organism of CR-BSI. This suggests that surviving strains carrying the mec A gene cause severe infection on empirical administration of an antibacterial drug, although biofilm formation by the ica AB gene is also important for CR-BSI. Based on these findings, most strains of S. epidermidis causing CR-BSI are biofilm-producing β-lactam-resistant (methicillin-resistant) bacteria. When S. epidermidis is isolated from blood culture testing, the identification of both ica AB and mec A genes may be significant with regard to judging whether the detected strain is the etiologic agent of CR-BSI. © Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases 2010.

Kidani T.,Ehime University | Kamei S.,Ehime University | Miyawaki J.,Ehime University | Aizawa J.,Ehime University | And 3 more authors.
Journal of Atherosclerosis and Thrombosis | Year: 2010

Aim: The aim of this study was to investigate whether environmental endocrine-disrupting chemicals, bisphenol A (BPA) and BPA-related chemicals, affect adiponectin production and secretion in 3T3-L1 adipocytes and whether BPA acts through Akt signaling. & Methods: 3T3-L1 adipocytes were treated for 24 h with BPA at various concentrations (20-80 μM) in serum-deprived medium. The medium was filtered through a 0.2 μm filter. Adiponectin in the infranatants of cell homogenates and in the media was measured using an adiponectin ELISA kit. The levels of Akt and p-Akt in cultures treated for 24 h with or without 80 μM BPA were analyzed by Western blot. &Results: The control cultures (i.e., BPA was absent during a 24-h treatment period) contained 49.4 μg/mg DNA of adiponectin in the cells and secreted 35.5 μg/mg DNA of adiponectin into the medium. BPA at 80 μM dose-dependently decreased the amounts of intracellular and medium adiponectin by 60% (p<0.01) and 56% (p<0.01), respectively, and decreased the levels of Akt and p-Akt by 46% (p <0.01) and 29% (p<0.01), respectively, compared with the control cultures. Like BPA, bisphenol F (BPF), bisphenol E (BPE), and bisphenol B (BPB) decreased the amounts of intracellular and medium adiponectin. The order of the potential to decrease the amount of intracellular adiponectin was BPB>BPA>BPE>BPF. & Conclusions: BPA downregulates Akt signaling and inhibits adiponectin production and secretion in 3T3-L1 adipocytes.

Kato N.,Ehime Prefectural University of Health Sciences
Bunseki Kagaku | Year: 2015

This study investigated the combustion conditions for Cr(VI) formation during pulverized coal combustion using three types of furnaces. Cr(VI) compounds can be highly soluble in water, whereas Cr(III) compounds are less soluble. The amount of Cr(VI) in coal ash was measured using the amount of Cr(VI) leached in a leaching solution, with ICP-AES after the separation of Cr(III) by Fe(OH)3 co-precipitation. The results from laboratory drop tube furnace experiments indicated that the amount of Cr(VI) leached from coal ash depended on the temperature of the furnace, the air ratio (amount of air used in combustion to the theoretical air demand), and the O2 concentration in the carrier gas. Since these dependences were similar to those for NOx emission, a two-stage combustion method used as a NOx reducing method was examined by using an experimental furnace with a burner for pulverized coal firing and a coal-fired power plant. The results showed that the amount of leached Cr(VI) decreased along with NOx as the two-stage combustion ratio increased. Low- NOx combustion conditions likely reduced the Cr(VI) formation in the combustion zone. © 2015 The Japan Society for Analytical Chemistry.

Kato N.,Ehime Prefectural University of Health Sciences
Bunseki Kagaku | Year: 2013

A study was conducted to allow the sorting of coal before import using only a small amount of coal by estimating the amount of leachable Cr(VI). The amount of leached Cr(VI) was determined in eight fly-ash samples from a power plant after extraction with dilute hydrochloric acid (pH 5.8 - 6.3). The Cr(VI)/total Cr ratio varied widely in the range of 0.25 to 5.0 % among the coal-ash samples tested. To prepare coal-ash similar to fly-ash from a coal-fired power plant, three coal types were burned in two types of combustion furnaces. One was an experimental furnace with a burner for pulverized coal firing. The leached Cr(VI) concentrations from coal-ash prepared using the experimental furnace were in good agreement with those from a coal-fired power plant, allowing estimates of leached Cr(VI) concentrations from coal-ash prepared in the experimental furnace. The second type was an electric furnace that incinerated pulverized coal on a ceramic dish under an air atmosphere at temperatures from room temperature to a preset value, according to the Japanese Industrial Standard. The Cr(VI) concentration from coal-ash prepared using the electric furnace depended on both the incinerating temperature and type of coal; it was not possible to select an incinerating temperature suitable for all types of coal-ash that had a leached Cr(VI) concentration, in good agreement with that from a coal-fired power plant. © 2013 The Japan Society for Analytical Chemistry.

Shimokawa T.,Ehime University | Nabeka H.,Ehime University | Yamamiya K.,Ehime University | Wakisaka H.,Ehime Prefectural University of Health Sciences | And 3 more authors.
Cell and Tissue Research | Year: 2013

Prosaposin (PSAP) is as a trophic factor and an activator protein for sphingolipid hydrolase in lysosomes. We generated a specific antibody to PSAP and examined the spatiotemporal distribution of PSAP-immunoreactive (PSAP-IR) cells in the lymphatic tissues of Wistar rats. Immunoblots of tissue homogenates separated electrophoretically showed a single band for PSAP in brain but two bands in spleen. PSAP-IR cells were distributed in both the red and white pulp of the spleen, in both the cortex and medulla of the thymus and in mesenteric lymph nodes. Many PSAP-IR cells were found in the dome portion of Peyer's patches and the number of PSAP-IR cells increased with the age of the rat. To identify the PSAP-IR cells, double- and triple-immunostainings were performed with antibodies against PSAP, CD68 and CD1d. The large number of double- and triple-positive cells suggested that antigen-presenting cells contained much PSAP in these lymphatic tissues. Intense expression of PSAP mRNA, examined by in situ hybridisation, was observed in the red pulp and corona of the spleen. In rats, the PSAP gene generates two alternative splicing forms of mRNA: Pro+9 containing a 9-base insertion and Pro+0 without the insertion. We examined the expression patterns of the alternative splicing forms of PSAP mRNA in the spleen. The presence of both types of mRNA (Pro+9 and Pro+0) indicated that the spleen contains various types of prosaposin-producing and/or secreting cells. These findings suggest diverse functions for PSAP in the immune system. © 2013 Springer-Verlag Berlin Heidelberg.

PubMed | Ehime Prefectural University of Health Sciences
Type: Journal Article | Journal: Cells | Year: 2017

The linker of nucleoskeleton and cytoskeleton (LINC) complex is composed of the outer and inner nuclear membrane protein families Klarsicht, Anc-1, and Syne homology (KASH), and Sad1 and UNC-84 (SUN) homology domain proteins. Increasing evidence has pointed to diverse functions of the LINC complex, such as in nuclear migration, nuclear integrity, chromosome movement and pairing during meiosis, and mechanotransduction to the genome. In metazoan cells, the nuclear envelope possesses the nuclear lamina, which is a thin meshwork of intermediate filaments known as A-type and B-type lamins and lamin binding proteins. Both of lamins physically interact with the inner nuclear membrane spanning SUN proteins. The nuclear lamina has also been implicated in various functions, including maintenance of nuclear integrity, mechanotransduction, cellular signalling, and heterochromatin dynamics. Thus, it is clear that the LINC complex and nuclear lamins perform diverse but related functions. However, it is unknown whether the LINC complex-lamins interactions are involved in these diverse functions, and their regulation mechanism has thus far been elusive. Recent structural analysis suggested a dynamic nature of the LINC complex component, thus providing an explanation for LINC complex organization. This review, elaborating on the integration of crystallographic and biochemical data, helps to integrate this research to gain a better understanding of the diverse functions of the LINC complex.

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