Mitochondrial encephalopathy, lactic acidosis, and stroke-like episodes diagnosed after metformin-triggered stroke-like episodes.

A 40-year-old man with sensorineural hearing loss and diabetes mellitus was hospitalized with acute-onset impaired consciousness and clumsiness in his left hand. He had been taking metformin for 4 months. A neurological examination revealed confusion and weakness in the left upper limb. Increased lactate levels were detected in the serum and cerebrospinal fluid. Magnetic resonance imaging revealed lesions in the right parietal and bilateral temporal lobes with a lactate peak in magnetic resonance spectroscopy. Finally, we made a genetic diagnosis of mitochondrial encephalopathy, lactic acidosis, and stroke-like episodes based on the detection of m.3243A>G. It is well-known that metformin should not be administered in patients with mitochondrial encephalopathy, lactic acidosis, and stroke-like episodes because metformin inhibits mitochondrial function and triggers stroke-like episodes. However, our patient was diagnosed with mitochondrial encephalopathy, lactic acidosis, and stroke-like episodes after metformin administration. Thus, we encourage physicians to exercise caution in the prescription of metformin in patients with short stature, sensorineural hearing loss, or young-onset diabetes mellitus because these patients may have undiagnosed mitochondrial encephalopathy, lactic acidosis, and stroke-like episodes.

Mathematical model estimation of dengue fever transmission risk from Southeast and South Asia into Japan between 2016 and 2018.

BACKGROUND: Dengue fever is a viral infection transmitted to humans through the bite of a mosquito infected with the dengue virus. Dengue is one of the most common infectious diseases in the world, and its incidence is rapidly increasing. We estimated the risk of dengue importation from endemic countries to Japan and the transmission risk within Japan using data collected between 2016 and 2018. METHODS: We conducted simulations that included the number of reported dengue infections and travelers per month in ten countries in Southeast and South Asia. RESULTS: The estimated importation risks for Japanese returnees and international travelers from each of the ten endemic countries was approximately 1.0 every month from 2016 to 2018. The autochthonous transmission risk in Japan from any target country was 1.0 from June to September yearly. The estimated number of Japanese dengue cases returning to Japan is approximately 25 times higher than that of imported cases reported in Japan. CONCLUSIONS: The risk of dengue importation into Japan can be sufficiently high. Attention should be paid to autochthonous transmission spread between June and September when mosquitoes are active in Japan. Estimates of seasonal risk variation from each dengue virus-endemic country can be used to inform preventive and control measures for dengue in Japan.

Elevated alanine aminotransferase and low aspartate aminotransferase/alanine aminotransferase ratio are associated with chronic kidney disease among middle-aged women: a cross-sectional study.

BACKGROUND: Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) to ALT ratio (AST/ALT ratio) have been shown to be related to non-alcoholic fatty liver disease or insulin resistance, which was associated with chronic kidney disease (CKD). However, it is unclear whether ALT and AST/ALT ratio are associated with CKD. In this study, we examined the relationship of ALT and AST/ALT ratio to CKD among middle-aged females in Japan. METHODS: The present study included 29,133 women aged 40 to 64 years who had an annual health checkup in Japan during April 2013 to March 2014. Venous blood samples were collected to measure ALT, AST, gamma-glutamyltransferase (GGT), and creatinine levels. In accordance with previous studies, ALT > 40 U/L and GGT > 50 U/L were determined as elevated, AST/ALT ratio < 1 was regarded as low, and CKD was defined as estimated glomerular filtration rate < 60 mL/min/1.73 m2 and/or proteinuria. Logistic regression model was used to calculate the odds ratio (OR) and 95% confidence interval (CI) for CKD. RESULTS: "Elevated ALT and elevated GGT" and "elevated ALT and non-elevated GGT" significantly increased the OR for CKD when compared with "non-elevated ALT and non-elevated GGT" (OR: 2.56, 95% CI: 2.10-3.12 and OR: 2.24, 95% CI: 1.81-2.77). Compared with "AST/ALT ratio ≥ 1 and non-elevated GGT", "AST/ALT ratio < 1 and elevated GGT" and "AST/ALT ratio < 1 and non-elevated GGT" significantly increased the OR for CKD (OR: 2.73, 95% CI: 2.36-3.15 and OR: 1.68, 95% CI: 1.52-1.87). These findings still remained after adjustment for confounders. CONCLUSIONS: Elevated ALT was associated with CKD regardless of GGT elevation. Moreover, low AST/ALT ratio was also associated with CKD independent of GGT elevation.

Small-Cell Lung Cancer Comorbid with Pulmonary Mycobacterium avium Infection: A Case Report.

BACKGROUND: Small-cell lung cancer (SCLC) rarely coexists with pulmonary Mycobacterium avium intracellular complex (MAC) infection. The key drug for SCLC treatment is etoposide, which is metabolized by cytochrome P-450 (CYP) 3A4. Meanwhile, the key drugs for pulmonary MAC infection are clarithromycin (CAM) and rifampicin (RFP), and their metabolism influences CYP3A4. Therefore, treatment of concurrent SCLC and pulmonary MAC infection is difficult, and to the best of our knowledge, no report of treatments for concurrent SCLC and pulmonary MAC infection has been published. Patient Concerns and Diagnoses: A 65-year-old man presented to our hospital with abnormal findings of chest computed tomography: (1) a hilar region nodule in the left lung and mediastinal lymphadenopathy and (2) a thick-walled cavity lesion in the right upper lobe of the lung. After further examinations, the former lesions were diagnosed as SCLC, cT4N3M0, stage IIIC and the latter as pulmonary MAC infection, fibrocavitary disease. INTERVENTIONS AND OUTCOMES: Concurrent treatment was conducted with discontinuation of CAM and RFP before and after etoposide administration. Specifically, intravenous cisplatin and etoposide were administered on day 1 and days 1-3, respectively, and CAM, RFP, and ethambutol (EB) were administered orally on days 6-22 every 4 weeks. Concurrent radiotherapy was added to the drug administration on days 1-27 of the first cycle. The chemotherapy was continued for 4 cycles, followed by continuation of CAM and RFP administration. EB was discontinued because of optic nerve disorder. The treatments were conducted completely and safely, and both of the SCLC lesions and the MAC lesion were improved. CONCLUSIONS: Treatments for concurrent SCLC and pulmonary MAC infection may be successfully conducted with discontinuation of CAM and RFP before and after etoposide administration.

Imidazole-Aided Native Chemical Ligation: Imidazole as a One-Pot Desulfurization-Amenable Non-Thiol-Type Alternative to 4-Mercaptophenylacetic Acid.

Various bioactive proteins have been synthesized by native chemical ligation (NCL) and its combination with subsequent desulfurization (e.g., conversion from Cys to Ala). In NCL, excess 4-mercaptophenylacetic acid (MPAA) is generally added to facilitate the reaction. However, co-elution of MPAA with the ligation product during preparative high-performance liquid chromatography sometimes reduces its usefulness. In addition, contamination of MPAA disturbs subsequent desulfurization. Here, we report for the first time that imidazole can be adopted as an alternative to MPAA in NCL using a peptide-alkylthioester. The efficiency of the imidazole-aided NCL (Im-NCL) is similar to that of traditional MPAA-aided NCL. As model cases, we successfully synthesized adiponectin(19-107) and [Ser(PO3 H2 )65 ]-ubiquitin using Im-NCL with a one-pot desulfurization.

Efficient one-pot synthesis of CXCL14 and its derivative using an N-sulfanylethylanilide peptide as a peptide thioester equivalent and their biological evaluation.

CXCL14 is a CXC-type chemokine that exhibits chemotactic activity for immature dendritic cells, activated macrophages, and activated natural killer cells. However, its specific receptor and signaling pathway remain obscure. Recently, it was reported that CXCL14 binds to CXCR4 with high affinity and inhibits CXCL12-mediated chemotaxis. Furthermore, the CXCL14 C-terminal α-helical region is important for binding to its receptor. In this context, we chemically synthesized CXCL14 and its derivative with a one-pot method using N-sulfanylethylanilide peptide as a thioester equivalent. The synthetic CXCL14 proteins possessed inhibitory activities to CXCL12-mediated chemotaxis comparable with that of recombinant CXCL14. Moreover, we proved that chemically biotinylated CXCL14 binds to CXCR4 on cells by flow cytometry analysis.

Highly sensitive detection of Epstein-Barr virus-infected cells by EBER flow FISH.

When Epstein-Barr virus (EBV) infection is suspected, identification of infected cells is important to understand the pathogenesis, determinine the treatment strategy, and predict the prognosis. We used the PrimeFlow™ RNA Assay Kit with a probe to detect EBV-encoded small RNAs (EBERs) and multiple surface markers, to identify EBV-infected cells by flow cytometry. We analyzed a total of 24 patients [11 with chronic active EBV disease (CAEBV), 3 with hydroa vacciniforme lymphoproliferative disorder, 2 with X-linked lymphoproliferative disease type 1 (XLP1), 2 with EBV-associated hemophagocytic lymphohistiocytosis, and 6 with posttransplant lymphoproliferative disorder (PTLD)]. We compared infected cells using conventional quantitative PCR methods and confirmed that infected cell types were identical in most patients. Patients with CAEBV had widespread infection in T and NK cells, but a small amount of B cells were also infected, and infection in patients with XLP1 and PTLD was not limited to B cells. EBV-associated diseases are believed to be complex pathologies caused by EBV infecting a variety of cells other than B cells. We also demonstrated that infected cells were positive for HLA-DR in patients with CAEBV. EBER flow FISH can identify EBV-infected cells with high sensitivity and is useful for elucidating the pathogenesis.

Regional assessment of marine and estuarine sediment toxicity in Southern California, USA.

Sediment toxicity was investigated at 222 stations in the Southern California Bight (SCB) during 2008. This represented the first time that assessment methods established by California's new Sediment Quality Objectives program were employed in a survey of this scale. The goal was to determine the extent and magnitude of sediment toxicity in the SCB, how toxicity compared among specific environments, and whether toxicity has changed over the last decade. Two toxicity tests were used: the 10-day amphipod whole sediment survival test with Eohaustorius estuarius and a 48-h embryo development test with the mussel Mytilus galloprovincialis exposed at the sediment-water interface. Less than 1 % of the area of the SCB was found to be toxic to the amphipod test. No toxicity was found in offshore stations, but 14 % of embayment areas were toxic to the amphipods. The mussel test identified 13 % of the embayment areas to be toxic. Estuary and marina locations had the greatest areal extent of toxicity for both tests. The two toxicity methods agreed that sediments were not toxic at over half of the stations tested. The mussel test showed a greater magnitude of response than the amphipod. Sediment toxicity was shown to have declined in both extent and magnitude from levels measured in 1998 and 2003.

Chemical synthesis of biologically active monoglycosylated GM2-activator protein analogue using N-sulfanylethylanilide peptide.

Going to SEA(lide): Total chemical synthesis of a 162-residue glycoprotein analogue of the monoglycosylated human GM2-activator protein (GM2AP) was achieved. Key steps were the use of N-sulfanylethylanilide (SEAlide) peptides in the kinetic chemical ligation synthesis of a large peptide fragment, and a convergent native chemical ligation for final fragment assembly.

Heterogeneous biological membranes regulate protein partitioning via fluctuating diffusivity.

Cell membranes phase separate into ordered Lo and disordered Ld domains depending on their compositions. This membrane compartmentalization is heterogeneous and regulates the localization of specific proteins related to cell signaling and trafficking. However, it is unclear how the heterogeneity of the membranes affects the diffusion and localization of proteins in Lo and Ld domains. Here, using Langevin dynamics simulations coupled with the phase-field (LDPF) method, we investigate several tens of milliseconds-scale diffusion and localization of proteins in heterogeneous biological membrane models showing phase separation into Lo and Ld domains. The diffusivity of proteins exhibits temporal fluctuations depending on the field composition. Increases in molecular concentrations and domain preference of the molecule induce subdiffusive behavior due to molecular collisions by crowding and confinement effects, respectively. Moreover, we quantitatively demonstrate that the protein partitioning into the Lo domain is determined by the difference in molecular diffusivity between domains, molecular preference of domain, and molecular concentration. These results pave the way for understanding how biological reactions caused by molecular partitioning may be controlled in heterogeneous media. Moreover, the methodology proposed here is applicable not only to biological membrane systems but also to the study of diffusion and localization phenomena of molecules in various heterogeneous systems.

Synthetic procedure for N-Fmoc amino acyl-N-sulfanylethylaniline linker as crypto-peptide thioester precursor with application to native chemical ligation.

N-sulfanylethylanilide (SEAlide) peptides 1, obtainable using Fmoc-based solid-phase peptide synthesis (Fmoc SPPS), function as crypto-thioesters in native chemical ligation (NCL), yielding a wide variety of peptides/proteins. Their acylating potential with N-terminal cysteinyl peptides 2 can be tuned by the presence or absence of phosphate salts, leading to one-pot/multifragment ligation, operating under kinetically controlled conditions. SEAlide peptides have already been shown to be promising for use in protein synthesis; however, a widely applicable method for the synthesis of N-Fmoc amino acyl-N-sulfanylethylaniline linkers 4, required for the preparation of SEAlide peptides, is unavailable. The present study addresses the development of efficient condensation protocols of 20 naturally occurring amino acid derivatives to the N-sulfanylethylaniline linker 5. N-Fmoc amino acyl aniline linkers 4 of practical use in NCL chemistry, except in the case of the proline- or aspartic acid-containing linker, were successfully synthesized by coupling of POCl(3)- or SOCl(2)-activated Fmoc amino acid derivatives with sodium anilide species 6, without accompanying racemization and loss of side-chain protection. Furthermore, SEAlide peptides 7 possessing various C-terminal amino acids (Gly, His, Phe, Ala, Asn, Ser, Glu, and Val) were shown to be of practical use in NCL chemistry.

Pediatric liver failure with massive sinusoidal infiltration of histiocytes.

Histiocytic neoplasms, such as Langerhans cell histiocytosis (LCH) and disseminated juvenile xanthogranuloma (JXG), can involve the liver and sometimes cause liver failure. We aimed to classify non-LCH histiocytic proliferating disorders that do not exhibit typical disseminated JXG histology. We examined four pediatric patients who presented with liver failure and splenomegaly. Two patients with liver cirrhosis without cholestasis underwent liver transplantation (LT). The other two patients presented with giant cell hepatitis causing neonatal/infantile acute liver failure (ALF). The infantile ALF patient also underwent LT. Liver dysfunction developed after LT in all three transplant cases and the grafts exhibited massive sinusoidal infiltration of histiocytes with hemophagocytosis, similar to the native liver. The neonatal ALF patient was treated with an LCH-type chemotherapy regimen, and is alive and well at 18 months. Infiltrating histiocytes were positive for CD68 and CD163, and negative for CD1a, CD207, and S-100 protein. The BRAF V600E mutation was not present. Liver histological findings were not consistent with conventional disseminated JXG or LCH, although the histological findings in other organs overlapped those of well-known histiocytic neoplasms. The histological and immunohistochemical findings of infiltrating histiocytes suggest that these four cases constituted a disseminated JXG-like systemic disease.

Using acoustic telemetry to quantify potential contaminant exposure of Vermilion Rockfish (Sebastes miniatus), Hornyhead Turbot (Pleuronichthys verticalis), and White Croaker (Genyonemus lineatus) at wastewater outfalls in southern California.

Contaminant Exposure Models (CEMs) were developed to predict population-level tissue contaminant concentrations in fishes by pairing sediment-bound contaminant concentrations (DDTs, PCBs) and fine-scale acoustic telemetry data from a habitat-associated species (Vermilion Rockfish, Sebastes miniatus), nomadic flatfish species (Hornyhead Turbot, Pleuronichthys verticalis), and nomadic benthic/midwater schooling species (White Croaker, Genyonemus lineatus) tagged near wastewater outfalls in southern California. Model results were compared to contaminant concentrations in tissue samples. The CEMs developed require further refinement before implementation into management efforts but may act as steppingstones to help shift primary monitoring methods away from the regular field collection of fish for tissue contaminant analyses and towards behavioral modeling and habitat mapping. We also developed Kernel Density Estimates that can be used by managers immediately to identify regions that contribute most to contaminant exposure in species of concern. Prioritizing remediation efforts in these areas are likely to be most effective at improving fish health.

Hepatic Steatosis Index and Chronic Kidney Disease among Middle-Aged Individuals: A Large-Scale Study in Japan.

BACKGROUND: Though nonalcoholic fatty liver disease (NAFLD) is related to chronic kidney disease (CKD), it is unclear whether the hepatic steatosis index (HSI), a screening tool for NAFLD, is related to CKD. The present study investigated the relationship between HSI and CKD among middle-aged individuals in Japan. METHODS: Subjects were adults (aged 40-64 years) who received an annual health checkup in Japan between April 2013 and March 2014. Height and weight were measured, and venous blood samples were obtained to determine alanine aminotransferase (ALT), aspartate aminotransferase (AST), and creatinine levels. HSI was calculated by the following formula: HSI = 8 × ALT/AST ratio + body mass index (+2, if diabetes; +2, if female). CKD was defined as an estimated glomerular filtration rate < 60 mL/min/1.73 m2 and/or urinary protein of ≥ (+). Logistic regression analysis was performed to estimate the odds ratio (OR) and its 95% confidence interval (CI) for CKD. RESULTS: Data of 94,893 adults were analyzed. Compared with men with an HSI < 30, men with 30 ≤ HSI ≤ 36 (OR: 1.50, 95% CI: 1.40-1.61) and HSI > 36 (OR: 2.14, 95% CI: 1.99-2.31) had significantly higher ORs for CKD. Moreover, there was a significant dose-response relationship between HSI and CKD (P for trend < 0.001). Even after adjusting for confounders, the significant results persisted. These findings in men were similar to those in women. CONCLUSIONS: This study showed that the HSI was associated with CKD among middle-aged adults in Japan. Additionally, a dose-response relationship of HSI to CKD was observed. The present study suggested that it might be useful to monitor the HSI among middle-aged individuals to detect CKD at an early stage.

Ecological effects of wastewater treatment modifications on infauna communities off the coast of Orange County, California.

The Orange County Sanitation District (OCSD) is a public agency that provides treatment of wastewater for residents in Orange County, California. The final effluent characteristics at OCSD have been altered in the last 18 years due to the ~20-fold increase of sodium hypochlorite usage for disinfecting waste streams from 2002 to 2010, implementation of the Ground Water Replenishment System (GWRS) in 2008, and conversion to full secondary treatment, together with a major reduction of sodium hypochlorite usage in 2011. This study analyzed infauna data gathered from 1994 to 2016 at the zone of initial dilution (ZID) and reference stations located on the San Pedro Shelf to assess the effects of these treatment process changes on biota in the receiving water. Impacts to the infauna community within the ZID were observed during the period of high chlorine usage, including the increased proportion of the pollution tolerant polychaete Capitella capitata complex from 6.2% in 2003 to nearly 60% in 2010, the lowest Infaunal Trophic Index score in 2009, and the highest Benthic Response Index score in 2006. The degradation of the infauna community in the ZID was correlated with chlorination and was coincidental with the initiation of the GWRS. The infauna community at the ZID recovered rapidly after the implementation of full secondary treatment as indicated by the significant reduction of the relative abundance of C. capitata complex from 59.8% in 2010 to less than 0.1% after 2012, and by the markedly improved community health index scores. This study demonstrated the composition and biointegrity of the infauna community at the ZID varied in response to changes in the wastewater treatment process. Caution should be exercised at wastewater treatment plants when relatively higher dosages of sodium hypochlorite are used over a multi-year period, as this may negatively impact aquatic biota in the receiving water.

Endogenous secretory RAGE but not soluble RAGE is associated with carotid atherosclerosis in type 1 diabetes patients.

Advanced glycation end-products (AGEs) and the receptor for AGEs (RAGE) system play an important role in the development of diabetic complications. The soluble form of RAGE (sRAGE) that potentially counteracts AGEs consists of several forms, including endogenous secretory RAGE (esRAGE; a splice variant of RAGE) and cleaved-type soluble RAGE derived from cell-surface RAGE. The aim of this study was to compare sRAGE and esRAGE directly in patients with type 1 diabetes. The associations of both total sRAGE and esRAGE with markers of glycaemic control and with carotid intima-media thickness (IMT) as a marker of atherosclerosis were examined in 130 type 1 diabetes patients (aged 23.6+/-4.9 years) and 22 age-matched non-diabetic subjects. IMT was inversely correlated with esRAGE (r=-0.254, p=0.0015) but neither with sRAGE nor subtracted soluble RAGE values (that is, circulating total sRAGE values - circulating esRAGE values). Furthermore, a stepwise multivariate regression analysis revealed that esRAGE (F=7.3), but not sRAGE, was a variable that interacted independently of IMT. It is likely that circulating sRAGE and esRAGE are distinct markers and that circulating esRAGE levels, but not sRAGE levels, are associated with the status of early-stage atherosclerosis.

The impact of new-onset diabetes on arterial stiffness after renal transplantation.

New-onset diabetes after renal transplantation (NODAT) is known to be a potent risk factor for cardiovascular events. We therefore investigated the incidence and risk factors for NODAT, and evaluated surrogate endpoints of atherosclerosis in Japanese patients with stable renal function after renal transplantation. Seventy-nine patients were enrolled in the study, and a 75 g oral glucose tolerance test (OGTT) was performed in subjects excluding patients with known NODAT. We evaluated the risk factors for NODAT and the degree of atherosclerosis, determined by brachial-ankle pulse wave velocity (baPWV), ankle-brachial blood pressure index (ABPI) and intima-media thickness (IMT) of the carotid artery. Eleven patients diagnosed as NODAT had significantly higher fasting plasma glucose before transplantation, blood pressure, and incidence of hepatitis C virus (HCV) infection than patients without NODAT. Multivariate regression analysis revealed that the independent determinant of NODAT was fasting plasma glucose pre-transplantation, HCV infection and systolic blood pressure. The baPWV in patients with NODAT was significantly higher compared to that in patients without NODAT. In addition, the independent determinant of baPWV evaluated by multivariate regression analysis was an increase in systolic blood pressure and age, and a decrease of adiponectin levels. In conclusion, we found that high fasting plasma glucose prior to transplantation, HCV infection and high blood pressure are risk factors for NODAT in Japanese patients after renal transplantation. Since NODAT patients have advanced arterial stiffness probably due to high blood pressure, strict control of blood pressure will be important for preventing the development of cardiovascular disease in NODAT.

Oxidative stress and the JNK pathway are involved in the development of type 1 and type 2 diabetes.

Failure of pancreatic beta-cells is the common characteristic of type 1 and type 2 diabetes. Type 1 diabetes mellitus is induced by destruction of pancreatic beta-cells which is mediated by an autoimmune mechanism and consequent inflammatory process. Various inflammatory cytokines and oxidative stress are produced during this process, which has been proposed to play an important role in mediating beta-cell destruction. The JNK pathway is also activated by such cytokines and oxidative stress, and is involved in beta-cell destruction. Type 2 diabetes is the most prevalent and serious metabolic disease, and beta-cell dysfunction and insulin resistance are the hallmark of type 2 diabetes. Under diabetic conditions, chronic hyperglycemia gradually deteriorates beta-cell function and aggravates insulin resistance. This process is called "glucose toxicity". Under such conditions, oxidative stress is provoked and the JNK pathway is activated, which is likely involved in pancreatic beta-cells dysfunction and insulin resistance. In addition, oxidative stress and activation of the JNK pathway are also involved in the progression of atherosclerosis which is often observed under diabetic conditions. Taken together, it is likely that oxidative stress and subsequent activation of the JNK pathway are involved in the pathogenesis of type 1 and type 2 diabetes.

Examining the role of estrogenic activity and ocean temperature on declines of a coastal demersal flatfish population near the municipal wastewater outfall of Orange County, California, USA.

Wastewater treatment plant effluent introduces a mixture of pollutants into marine environments; however, the impacts of chronic sublethal exposures on populations are often unclear. Presence of estrogenic agents in sediments and uptake of these compounds by demersal flatfishes has been reported at the Orange County Sanitation District (OCSD) wastewater outfall. Furthermore, estrogenic activity has been identified in male flatfish in the area, potentially contributing to observed population declines in the OCSD region. Rising ocean temperatures may further contribute to flatfish declines as relationships between temperature and abundance have been reported in the Southern California Bight. To investigate declines, sex ratios, condition factor, organ health indices, hormones, and vitellogenin were quantified in flatfish collected at OCSD outfall and reference sites. Additionally, historical temperature data was examined for trends with population abundances. Rather than being linked to estrogenic activity, results indicated that population declines were more correlated to increases in ocean temperature.