Clinical Evaluation of Rapid Diagnostic Test Kit Using the Polysaccharide as a Genus-Specific Diagnostic Antigen for Leptospirosis in Korea, Bulgaria, and Argentina

Leptospirosis, a zoonotic disease that is caused by many serovars which are more than 200 in the world, is an emerging worldwide disease. Accurate and rapid diagnostic tests for leptospirosis are a critical step to diagnose the disease. There are some commercial kits available for diagnosis of leptospirosis, but the obscurity of a species- or genus-specific antigen of pathogenic Leptospira interrogans causes the reduced sensitivity and specificity. In this study, the polysaccharide derived from lipopolysaccharide (LPS) of nonpathogenic Leptospira biflexa serovar patoc was prepared, and the antigenicity was confirmed by immunoblot and enzyme linked immunosorbent assay (ELISA). The performance of the rapid diagnostic test (RDT) kit using the polysaccharide as a diagnostic antigen was evaluated in Korea, Bulgaria and Argentina. The sensitivity was 93.9%, 100%, and 81.0% and the specificity was 97.9%, 100%, and 95.4% in Korea (which is a rare region occurring with 2 serovars mostly), Bulgaria (epidemic region with 3 serovars chiefly) and Argentina (endemic region with 19 serovars mainly) respectively. These results indicate that this RDT is applicable for global diagnosis of leptospirosis. This rapid and effective diagnosis will be helpful for diagnosis and manage of leptospirosis to use and the polysaccharide of Leptospira may be called as genus specific antigen for diagnosis

Amplification of Enterocytozoon hepatopenaei in Penaeus vannamei Hepatopancreas Primary Culture and Immunofluorescence Assay for Detection of Enterocytozoon hepatopenaei

Hepatopancreatic microsporidiosis (HPM) disease leads to retarded growth in shrimp resulting in a major loss for the shrimp industry worldwide. The causative agent of HPM is a microsporidian known as Enterocytozoon hepatopenaei (EHP). It is little understood how EHP infects its host and hijacks its cellular machinery to replicate more organisms. Lack of an immortal cell line is a bottleneck in studying the cellular and molecular basis of EHP infection in shrimp. For this reason, EHP cannot be propagated in in vitro culture and must be propagated in live shrimp. The use of live EHP-infected shrimp remains the only way to study EHP infectivity. It was hypothesized that supplementing EHP with fresh host cells will aid the propagation of EHP in vitro. Further research must be done but with the data collected at this point, this hypothesis is rejected. In addition to the challenges in amplifying EHP in in vitro culture, there is no antibody-based detection method for EHP. EHP infection in a shrimp is examined by Hematoxylin and Eosin (H&E) histology and real-time polymerase chain reaction based detection methods. Monoclonal antibodies that were previously characterized by Riggs and colleagues to detect Cryptosporidium parvum (C. parvum), successfully detected other parasites. Based on this, it was hypothesized that monoclonal antibodies against C. parvum may also detect EHP

Deep reinforcement learning in an ultrafiltration system: Optimizing operating pressure and chemical cleaning conditions

Enhancing engineering efficiency and reducing operating costs are permanent subjects that face all engineers over the world. To effectively improve the performance of filtration systems, it is necessary to determine an optimal operating condition beyond conventional methods of periodic and empirical operation. Herein, this paper proposes an effective approach to finding an optimal operating strategy using deep reinforcement learning (DRL), particularly for an ultrafiltration (UF) system. Deep learning was developed to represent the UF system utilizing a long-short term memory and provided an environment for DRL. DRL was designed to control three actions; operating pressure, cleaning time, and cleaning concentration. Ultimately, DRL proposed the UF system to actively change the operating pressure and cleaning conditions over time toward better water productivity and operating efficiency. DRL denoted similar to 20.9% of specific energy consumption can be reduced by increasing average water flux (39.5-43.7 L m(-2) h(-1)) and reducing operating pressure (0.617-0.540 bar). Moreover, the optimal action of DRL was reasonable to achieve better performance beyond the conventional operation. Crucially, this study demonstrated that due to the nature of DRL, the approach is tractable for engineering systems that have structurally complex relationships among operating conditions and resultants

Rho-kinase/AMPK axis regulates hepatic lipogenesis during overnutrition

This work was supported by grants from the NIH (R01DK083567 to YBK), the American Diabetes Association (1-09-RA-87 to YBK), the American Heart Association (12GRANT12040170 to YBK), the East Carolina University Start-up fund (to HH), the National Research Foundation (NRF-2014M3A9D8034464 to M Shong, NRF-2016R1A2B3010373 to KSP, NRF-2015R1C1A1A02037164 to SHL), the National Research Foundation of Korea (2013M3C7A1056024 to MSK), and the Korean Diabetes Association (to JAS, 2017). In addition, structural funding for the Center for Neuroscience and Cell Biology, University of Coimbra, NMR facility is supported by FEDER-PT2020 (UID/BIA/04004/2013 and CENTRO-07-CT62-FEDER-002012) and by the Portuguese Foundation for Science and Technology (FCT) through grants PTDC/CVT-NUT/2851/2014, PTDC/BIM-MET/4265/2014, and RECI/QEQ-QFI/0168/2012. ISL is a recipient of an FCT fellowship from Portugal (SFRH/BD/71021/2010), and MCK is a recipient of a postdoctoral fellowship award from the American Diabetes Association (1-17-PDF-146). GDB is supported by the European Union's Horizon 2020 Research and Innovation programme under Marie Sklodowska-Curie Grant Agreement 722619. We thank Farhad Danesh for CA-ROCK1-knockin mice; Huseyin Ozkan, Ivan Viegas, Cristina Barosa, Hyun Cheol Rho, Xuemei Ma, Yao Yang, and Alexander Banks for technical help; and Barbara Kahn, Tony Hollenberg, Sonia Najjar, and Terry Flier for helpful discussion.Obesity is a major risk factor for developing nonalcoholic fatty liver disease (NAFLD). NAFLD is the most common form of chronic liver disease and is closely associated with insulin resistance, ultimately leading to cirrhosis and hepatocellular carcinoma. However, knowledge of the intracellular regulators of obesity-linked fatty liver disease remains incomplete. Here we showed that hepatic Rho-kinase 1 (ROCK1) drives obesity-induced steatosis in mice through stimulation of de novo lipogenesis. Mice lacking ROCK1 in the liver were resistant to diet-induced obesity owing to increased energy expenditure and thermogenic gene expression. Constitutive expression of hepatic ROCK1 was sufficient to promote adiposity, insulin resistance, and hepatic lipid accumulation in mice fed a high-fat diet. Correspondingly, liver-specific ROCK1 deletion prevented the development of severe hepatic steatosis and reduced hyperglycemia in obese diabetic (ob/ob) mice. Of pathophysiological significance, hepatic ROCK1 was markedly upregulated in humans with fatty liver disease and correlated with risk factors clustering around NAFLD and insulin resistance. Mechanistically, we found that hepatic ROCK1 suppresses AMPK activity and a ROCK1/AMPK pathway is necessary to mediate cannabinoid-induced lipogenesis in the liver. Furthermore, treatment with metformin, the most widely used antidiabetes drug, reduced hepatic lipid accumulation by inactivating ROCK1, resulting in activation of AMPK downstream signaling. Taken together, our findings establish a ROCK1/AMPK signaling axis that regulates de novo lipogenesis, providing a unique target for treating obesity-related metabolic disorders such as NAFLD.publishersversionpublishe