Arabidopsis ETO1 specifically interacts with and negatively regulates type 2 1-aminocyclopropane-1-carboxylate synthases

BACKGROUND: In Arabidopsis, ETO1 (ETHYLENE-OVERPRODUCER1) is a negative regulator of ethylene evolution by interacting with AtACS5, an isoform of the rate-limiting enzyme, 1-aminocyclopropane-1-carboxylate synthases (ACC synthase or ACS), in ethylene biosynthetic pathway. ETO1 directly inhibits the enzymatic activity of AtACS5. In addition, a specific interaction between ETO1 and AtCUL3, a constituent of a new type of E3 ubiquitin ligase complex, suggests the molecular mechanism in promoting AtACS5 degradation by the proteasome-dependent pathway. Because orthologous sequences to ETO1 are found in many plant species including tomato, we transformed tomato with Arabidopsis ETO1 to evaluate its ability to suppress ethylene production in tomato fruits. RESULTS: Transgenic tomato lines that overexpress Arabidopsis ETO1 (ETO1-OE) did not show a significant delay of fruit ripening. So, we performed yeast two-hybrid assays to investigate potential heterologous interaction between ETO1 and three isozymes of ACC synthases from tomato. In the yeast two-hybrid system, ETO1 interacts with LE-ACS3 as well as AtACS5 but not with LE-ACS2 or LE-ACS4, two major isozymes whose gene expression is induced markedly in ripening fruits. According to the classification of ACC synthases, which is based on the C-terminal amino acid sequences, both LE-ACS3 and AtACS5 are categorized as type 2 isozymes and possess a consensus C-terminal sequence. In contrast, LE-ACS2 and LE-ACS4 are type 1 and type 3 isozymes, respectively, both of which do not possess this specific C-terminal sequence. Yeast two-hybrid analysis using chimeric constructs between LE-ACS2 and LE-ACS3 revealed that the type-2-ACS-specific C-terminal tail is required for interaction with ETO1. When treated with auxin to induce LE-ACS3, seedlings of ETO1-OE produced less ethylene than the wild type, despite comparable expression of the LE-ACS3 gene in the wild type. CONCLUSION: These results suggest that ETO1 family proteins specifically interact with and negatively regulate type 2 ACC synthases. Our data also show that Arabidopsis ETO1 can regulate type 2 ACS in a heterologous plant, tomato

Staging the tumor and staging the host: A two centre, two country comparison of systemic inflammatory responses of patients undergoing resection of primary operable colorectal cancer

Background: How systemic inflammation-based prognostic scores such as the modified Glasgow Prognostic Score (mGPS) and neutrophil:lymphocyte ratio (NLR) differ across populations of patients with colorectal cancer (CRC) remains unknown. The present study examined the mGPS and NLR in patients from United Kingdom (UK) and Japan. Methods: Patients undergoing resection of TNM I-III CRC in two centres in the UK and Japan were included. Differences in clinicopathological characteristics and mGPS (0-CRP≤10 mg/L, 1-CRP>10 mg/L, 2-CRP>10 mg/L, albumin<35 g/L) and NLR (≤5/>5) were examined. Results: Patients from UK (n = 581) were more likely to be female, high ASA and BMI, present as an emergency (all P < 0.01) and have higher T stage compared to those from Japan (n = 559). After controlling for differences in tumor and host characteristics, patients from Japan were less likely to be systemically inflamed (OR: mGPS: 0.37, 95%CI 0.27–0.50, P < 0.001; NLR: 0.53, 95%CI 0.35–0.79, P = 0.002). Conclusion: Systemic inflammatory responses differ between populations with colorectal cancer. Given their prognostic value, reporting of systemic inflammation-based scores should be incorporated into future studies reporting patient outcomes. Summary: Although the systemic inflammatory response is recognised as a prognostic factor in patients with colorectal cancer, it is not clear how these may differ between distinct geographical populations. The present study examines differences in the prevalence of elevated systemic inflammatory responses (modified Glasgow Prognostic Score and neutrophil:lymphocyte ratio) between two populations undergoing resection of colorectal cancer in the United Kingdom and Japan

Clinical features and pathogenesis of membranoproliferative glomerulonephritis: a nationwide analysis of the Japan renal biopsy registry from 2007 to 2015

BackgroundThe incidence and age distribution of membranoproliferative glomerulonephritis (MPGN) vary throughout the world by race and ethnicity. We sought to evaluate the clinical features, pathogenesis, and age distribution of MPGN among a large nationwide data from the Japan Renal Biopsy Registry (J-RBR).MethodsA cross-sectional survey of 593 patients with MPGN (types I and III) registered in the J-RBR between 2007 and 2015 was conducted. Clinical parameters, and laboratory findings at diagnosis were compared between children (< 20 years), adults (20–64 years), and elderly patients (≥ 65 years).ResultsThe median age of the patients was 59.0 years and mean proteinuria was 3.7 g/day. The rate of nephrotic syndrome was significantly higher in adults (40.4%) and elderly patients (54.0%) than in children (14.9%), whereas the rate of chronic glomerulonephritis was significantly higher in children (66.2%) than in adults (34.4%) and elderly patients (31.2%). According to the CGA risk classification, high-risk (red zone) cases accounted for 3.4% of children, 52.5% of adults and 84.1% of elderly patients with MPGN. As for pathogenesis, primary MPGN was most frequent (56.0%). Lupus nephritis was the most common disease among adult patients with secondary MPGN, whereas infectious disease was more common in elderly patients. Multiple regression analysis revealed that high systolic blood pressure and high proteinuria were independent factors associated with decreased estimated glomerular filtration rate (eGFR) in adults and elderly patients with MPGN.ConclusionsIn Japan, adults and elderly patients with MPGN had a lower eGFR and severer proteinuria than children

CARRIER-MEDIATED UPTAKE OF H 2 -RECEPTOR ANTAGONISTS BY THE RAT CHOROID PLEXUS: INVOLVEMENT OF RAT ORGANIC ANION TRANSPORTER 3

ABSTRACT: The choroid plexus (CP) acts as a site for the elimination of xenobiotic organic compounds from the cerebrospinal fluid (CSF). The purpose of the present study is to investigate the role of rat organic anion transporter 3 (rOat3; Slc22a8) in the uptake of H 2 -receptor antagonists (cimetidine, ranitidine, and famotidine) by the isolated rat CP. Saturable uptake of cimetidine and ranitidine was observed in rOat3-LLC with K m values of 80 and 120 M, respectively, whereas famotidine was found to be a poor substrate. The steady-state concentration of the H 2 -receptor antagonists in the CSF was significantly increased by simultaneously administered probenecid, although it did not affect their brain and plasma concentrations. Saturable uptake of cimetidine and ranitidine was observed in the isolated rat CP with K m values of 93 and 170 M, respectively, whereas 50% of the uptake of famotidine remained at the highest concentration examined (1 mM). The K i value of ranitidine for the uptake of cimetidine by the isolated CP (50 M) was similar to its own K m value, suggesting that they share the same transporter for their uptake. The inhibition potency of organic anions such as benzylpenicillin, estradiol 17␤-glucuronide, p-aminohippurate, and estrone sulfate for the uptake of cimetidine by the isolated rat CP was similar to that for benzylpenicillin, the uptake of which has been hypothesized to be mediated by rOat3, whereas a minimal effect by tetraethylammonium excludes involvement of organic cation transporter(s). These results suggest that rOat3 is the most likely candidate transporter involved in regulating the CSF concentration of H 2 -receptor antagonists at the CP

Retrorectal epidermoid cyst with unusually elevated serum SCC level, initially diagnosed as an ovarian tumor

Retrorectal epidermoid cyst is one of the developmental cysts which arise from remnants of embryonic tissues. We report a rare case of retrorectal epidermoid cyst, initially diagnosed as an ovarian tumor. Serum SCC value as tumor marker was elevated to the high level. Laparoscopy revealed ovaries, uterus and other pelvic organs were all normal. This tumor existed in the retroperitoneal cavity and compressed the rectum. Later, complete tumor resection was performed by laparotomy. Histological study revealed the epithelium of this tumor consisted of only squamous cells without atypia, and the diagnosis of this tumor was retrorectal epidermoid cyst. Retrorectal epidermoid cyst is very rare, and difficult to diagnose before surgery. However, if we have-knowledge of developmental cysts, and by careful digital examination and image diagnosis, a differential diagnosis can be made

Drug-induced kidney disease: a study of the Japan Renal Biopsy Registry from 2007 to 2015

IntroductionThe Japan Renal Biopsy Registry (J-RBR) was started in 2007 by the Committee for the Standardization of Renal Pathological Diagnosis and the Committee for the Kidney Disease Registry of the Japanese Society of Nephrology. The purpose of this report is to clarify drug-induced kidney disease (DIKD) of renal biopsied cases in Japan.Subjects and methodsWe analyzed the data of 26,535 cases that were registered in the J-RBR from 2007 to 2015.ResultsBased on clinical and pathological diagnoses, 328 cases (176 males and 152 females) of renal biopsy-proven DIKD were registered in the J-RBR from 2007 to 2015 (1.24 % of all cases). The frequency of DIKD increased with age. The number of cases peaked in the 6th–8th decade in all pathological categories, except for the number of chronic tubulointerstitial lesions (CTIL), which peaked in the 4th–5th decade. Overall, the frequency of DIKD was 3 times higher in the 7th decade than in the 2nd decade (1.86 vs. 0.62 %). The main clinical diagnoses were DIKD in 150 cases (45.7 %), nephrotic syndrome in 66 cases (20.1 %), chronic nephritic syndrome in 55 cases (16.8 %), and rapidly progressive glomerulonephritis in 30 cases (9.1 %). DIKD was registered as a secondary diagnosis in 136 cases (41.5 %). The pathological findings of these cases were glomerular lesions in 105 cases (32.0 %), acute tubulointerstitial lesions (ATIL) in 87 cases (26.5 %), CTIL in 72 cases (22.0 %), and sclerotic glomerular lesions and/or nephrosclerosis in 18 cases (5.5 %). ATIL and CTIL were mainly found in cases in which DIKD was diagnosed on the basis of the patient’s clinical findings. In addition, nephrotic syndrome-related membranous nephropathy (MN) was the major cause of renal damage in 59.4 % of the cases involving glomerular injuries. According to the CGA risk classification, high-risk (red zone) cases accounted for 56.1 % of all cases of DIKD and 75.9, 64.9, and 33.3 % of the cases involving ATIL, CTIL, and glomerular injuries, respectively. The causative drugs were identified in 102 cases, including bucillamine in 38 cases of MN, gemcitabine in 3 cases of thrombotic microangiopathy, and other anticancer drugs in 14 cases (anti-vascular endothelial growth factor drugs in 3 cases and propyl thiouracil in 3 cases of anti-neutrophil cytoplasmic antibody-related nephritis).ConclusionOur analysis of the J-RBR revealed that DIKD mainly affects elderly people in Japan. ATIL or CTIL were found in approximately half of the biopsied cases of DIKD, and one-third involved glomerular lesions, mainly MN or clinical nephrotic syndrome

Morphological Alterations of the Eccrine Sweat Apparatus in Amputated Feet from Diabetes Mellitus Patients

Several physiological studies have demonstrated decreased or absent thermoregulatory sweating in the distal legs and feet of diabetes patients. Such hypohidrosis in diabetes patients is believed to be a clinical symptom of autonomic neuropathy. Thus, the present study sought to clarify the pathogenesis of structural alterations of the eccrine sweat apparatus in diabetes patients. For this study, we enrolled 17 patients with diabetic ulcers/gangrene who underwent amputation of the foot. Specimens were obtained 30mm from the ulcer/gangrene after amputation using a 6-mm trepan, with 12 normal human skin samples obtained from areas adjacent to pigmented nevi or benign skin tumors on the legs or feet to serve as controls. Numbers and morphological abnormalities of eccrine sweat glands and ducts were assessed by light microscopy. The pathogenesis of morphological alterations was examined by electron microscopy and immunoelectron microscopy of type IV collagen. Rates of disappearance of the lumen, shrunken morphology, and irregular outlines of eccrine sweat glands and ducts were significantly higher or more abundant in diabetes patients than in controls (P = 0.0002〜0.0001). Ultrastructurally, we observed prominent thickening of the basement membranes in eccrine sweat glands, admixed cell debris, and narrowing of the lumenal space. The thickened basement membranes resulted in the shrunken morphology and irregular outlines in eccrine sweat glands and ducts. Immunoelectron microscopy showed immunogold labeling for type IV collagen throughout the thickened basement membrane zone. These morphological alterations of the eccrine sweat apparatus in amputated feet from diabetes patients could be caused by diabetic and/or uremic neuropathy, and at least in part by angiopathy

X-Ray and Near-Infrared Observations of GX 339-4 in the Low/Hard State with Suzaku and IRSF

X-ray and near-infrared ($J$-$H$-$K_{\rm s}$) observations of the Galactic black hole binary GX 339--4 in the low/hard state were performed with Suzaku and IRSF in 2009 March. The spectrum in the 0.5--300 keV band is dominated by thermal Comptonization of multicolor disk photons, with a small contribution from a direct disk component, indicating that the inner disk is almost fully covered by hot corona with an electron temperature of $\approx$175 keV. The Comptonizing corona has at least two optical depths, $\tau \approx 1,0.4$. Analysis of the iron-K line profile yields an inner disk radius of $(13.3^{+6.4}_{-6.0}) R_{\rm g}$ ($R_{\rm g}$ represents the gravitational radius $GM/c^2$), with the best-fit inclination angle of $\approx50^\circ$. This radius is consistent with that estimated from the continuum fit by assuming the conservation of photon numbers in Comptonization. Our results suggest that the standard disk of GX 339--4 is likely truncated before reaching the innermost stable circular orbit (for a non rotating black hole) in the low/hard state at $\sim$1% of the Eddington luminosity. The one-day averaged near-infrared light curves are found to be correlated with hard X-ray flux with $F_{\rm Ks} \propto F_{\rm X}^{0.45}$. The flatter near infrared $\nu F_{\nu}$ spectrum than the radio one suggests that the optically thin synchrotron radiation from the compact jets dominates the near-infrared flux. Based on a simple analysis, we estimate the magnetic field and size of the jet base to be $5\times10^4$ G and $6\times 10^8$ cm, respectively. The synchrotron self Compton component is estimated to be approximately 0.4% of the total X-ray flux.Comment: 17pages, 15 figures, accepted for publication in PASJ (Suzaku and MAXI Special Issue

The role of infection in the development of non-valvular atrial fibrillation: Up-regulation of Toll-like receptor 2 expression levels on monocytes

SummaryMany studies have suggested that inflammation may participate in the pathogenesis of non-valvular atrial fibrillation (AF). However, it has been unknown by exposure to what the inflammation is caused. Recently, we reported that Toll-like receptor 2 (TLR2) level on monocytes was significantly up-regulated in viral and bacterial infections, but not in non-infectious inflammatory states. Our purpose was to test the hypothesis that expression of TLR2 levels may be up-regulated in patients with non-valvular AF. A total of 48 consecutive patients with non-valvular AF who were hospitalized for catheter ablation were enrolled in this study. TLR2 levels were assayed by using flow-cytometric analysis and compared with volunteers in sinus rhythm (control group, n=24). Additionally, C-reactive protein (CRP) and interleukin-6 (IL-6) levels were assayed, and the left atrial volume indexes (LAVI) in the non-valvular AF group were measured. The results demonstrated that TLR2 levels in the non-valvular AF group were significantly higher than in the control group (median, 4682 vs. 3866 sites/cell; P<0.01). Moreover, non-valvular AF patients had significantly higher IL-6 levels than controls. However, there was no significant difference in CRP levels between the two groups. It was observed in 44 AF patients, in whom pulmonary vein isolation was confirmed to be successful, that the LAVI significantly diminished 1 month after ablation (median, 33.6 vs. 29.5ml/m2; P<0.001), but not the TLR2 and IL-6 levels. Our results implied that an infectious inflammation may participate in the pathogenesis of non-valvular AF