Endoscopic activity in inflammatory bowel disease: clinical significance and application in practice

Endoscopy is vital for diagnosis, assessing treatment response, monitoring and surveillance in patients with inflammatory bowel disease (IBD). With the growing importance of mucosal healing as a treatment target, the assessment of disease activity by endoscopy has been accepted as the standard of care for IBD. There are many endoscopic activity indices for facilitating standardized reporting of the gastrointestinal mucosal appearance in IBD, and each index has its strengths and weaknesses. Although most endoscopic indices do not have a clear-cut validated definition, endoscopic remission or mucosal healing is associated with favorable outcomes, such as a decreased risk of relapse. Therefore, experts suggest utilizing endoscopic indices for monitoring disease activity and optimizing treatment to achieve remission. However, the regular monitoring of endoscopic activity is limited in practice owing to several factors, such as the complexity of the procedure, time consumption, inter-observer variability, and lack of a clear-cut, validated definition of endoscopic response or remission. Although experts have recently suggested consensus-based definitions, further studies are needed to define the values that can predict long-term outcomes

Fecal Microbiota Transplantation: An Update on Clinical Practice

Fecal microbiota transplantation (FMT) is an infusion in the colon, or the delivery through the upper gastrointestinal tract, of stool from a healthy donor to a recipient with a disease believed to be related to an unhealthy gut microbiome. FMT has been successfully used to treat recurrent Clostridium difficile infection (rCDI). The short-term success of FMT in rCDI has led to investigations of its application to other gastrointestinal disorders and extra-intestinal diseases with presumed gut dysbiosis. Despite the promising results of FMT in these conditions, several barriers remain, including determining the characteristics of a healthy microbiome, ensuring the safety of the recipient with respect to long-term outcomes, adequate monitoring of the recipient of fecal material, achieving high-quality control, and maintaining reasonable costs. For these reasons, establishing uniform protocols for stool preparation, finding the best modes of FMT administration, maintaining large databases of donors and recipients, and assuring that oral ingestion is equivalent to the more widely accepted colonoscopic infusion are issues that need to be addressed

Nucleotidylylation of the VPg protein of a human norovirus by its proteinase-polymerase precursor protein

AbstractCaliciviruses have a positive strand RNA genome covalently-linked at the 5'-end to a small protein, VPg. This study examined the biochemical modification of VPg by the ProPol form of the polymerase of human norovirus strain MD145 (GII.4). Recombinant norovirus VPg was shown to be nucleotidylylated in the presence of Mn2+ by MD145 ProPol. Phosphodiesterase I treatment of the nucleotidylylated VPg released the incorporated UMP, which was consistent with linkage of RNA to VPg via a phosphodiester bond. Mutagenesis analysis of VPg identified Tyrosine 27 as the target amino acid for this linkage, and suggested that VPg conformation was important for the reaction. Nucleotidylylation was inefficient in the presence of Mg2+; however the addition of full- and subgenomic-length MD145 RNA transcripts led to a marked enhancement of the nucleotidylylation efficiency in the presence of this divalent cation. Furthermore, evidence was found for the presence of an RNA element near the 3'-end of the polyadenylated genome that enhanced the efficiency of nucleotidylylation in the presence of Mg2+

Stable expression of a Norwalk virus RNA replicon in a human hepatoma cell line

AbstractNorwalk virus (NV) is a prototype strain of the genus Norovirus in the family Caliciviridae. The human noroviruses have emerged as major agents of acute gastroenteritis in all age groups, but there are no vaccines or antiviral agents partly due to the absence of a cell culture system. We report the generation of cells expressing self-replicating NV RNA (NV replicon) following transfection of NV RNA bearing an engineered neomycin resistance gene into cell lines of human (Huh-7) or hamster (BHK21) origin. Expression of replicon RNA was significantly reduced in the presence of interferon (IFN)-α in a dose-dependent manner in the NV replicon-bearing cells, suggesting a role for innate immunity in the control of human norovirus replication. This stable NV replicon system should lead to new insights into norovirus replication, virus–host interactions, and approaches for the treatment of norovirus disease

Structural and Inhibitor Studies of Norovirus 3C-like Proteases

Noroviruses have a single-stranded, positive sense 7–8 kb RNA genome, which encodes a polyprotein precursor processed by a virus-encoded 3C-like cysteine protease (3CLpro) to generate mature non-structural proteins. Because processing of the polyprotein is essential for virus replication, norovirus 3CLpro has been targeted for the discovery of anti-norovirus small molecule therapeutics. Thus, we performed functional, structural and inhibition studies of norovirus 3CLpro with fluorescence resonance energy transfer (FRET) assay, X-ray crystallography, and NMR spectroscopy with a synthetic protease inhibitor. Three 3CLpro from Norwalk virus (NV, genogroup I), MD145 (genogroup II) and murine norovirus-1 (MNV-1, genogroup V) were optimized for a FRET assay, and compared for the inhibitory activities of a synthetic protease inhibitor (GC376). The apo 3D structures of NV 3CLpro determined with X-ray crystallography and NMR spectroscopy were further analyzed. In addition, the binding mode of NV 3CLpro-GC376 was compared with X-ray crystallography and NMR spectroscopy. The results of this report provide insight into the interaction of NV 3CLpro with substrate/inhibitor for better understanding of the enzyme and antiviral drug development

Synthesis and anti-norovirus activity of pyranobenzopyrone compounds

During the last decade, noroviruses have gained media attention as the cause of large scale outbreaks of gastroenteritis on cruise ships, dormitories, nursing homes, etc. Although noroviruses do not multiply in food or water, they can cause large outbreaks because approximately 10–100 virions are sufficient to cause illness in a healthy adult. Recently, it was shown that the activity of acyl-coenzyme A:cholesterol acyltransferase-1 (ACAT1) enzyme may be important in norovirus infection. In search of anti-noroviral agents based on the inhibition of ACAT1, we synthesized and evaluated the inhibitory activities of a class of pyranobenzopyrone molecules containing amino, pyridine, substituted quinolines, or 7,8-benzoquinoline nucleus. Three of the sixteen evaluated compounds possess ED[subscript]5[subscript]0 values in the low micrometer range. 2-Quinolylmethyl derivative 3A and 4-quinolylmethyl derivative 4A showed ED[subscript]5[subscript]0 values of 3.4 and 2.4 [mu]M and TD[subscript]5[subscript]0 values of >200 and 96.4 [mu]M, respectively. The identified active compounds are suitable for further modification for the development of anti-norovirus agents

Reversal of the Progression of Fatal Coronavirus Infection in Cats by a Broad-Spectrum Coronavirus Protease Inhibitor

Citation: Kim Y, Liu H, Galasiti Kankanamalage AC, Weerasekara S, Hua DH, Groutas WC, et al. (2016) Reversal of the Progression of Fatal Coronavirus Infection in Cats by a Broad-Spectrum Coronavirus Protease Inhibitor. PLoS Pathog 12(3): e1005531. doi:10.1371/journal.ppat.1005531Coronaviruses infect animals and humans causing a wide range of diseases. The diversity of coronaviruses in many mammalian species is contributed by relatively high mutation and recombination rates during replication. This dynamic nature of coronaviruses may facilitate cross-species transmission and shifts in tissue or cell tropism in a host, resulting in substantial change in virulence. Feline enteric coronavirus (FECV) causes inapparent or mild enteritis in cats, but a highly fatal disease, called feline infectious peritonitis (FIP), can arise through mutation of FECV to FIP virus (FIPV). The pathogenesis of FIP is intimately associated with immune responses and involves depletion of T cells, features shared by some other coronaviruses like Severe Acute Respiratory Syndrome Coronavirus. The increasing risks of highly virulent coronavirus infections in humans or animals call for effective antiviral drugs, but no such measures are yet available. Previously, we have reported the inhibitors that target 3C-like protease (3CLpro) with broad-spectrum activity against important human and animal coronaviruses. Here, we evaluated the therapeutic efficacy of our 3CLpro inhibitor in laboratory cats with FIP. Experimental FIP is 100% fatal once certain clinical and laboratory signs become apparent. We found that antiviral treatment led to full recovery of cats when treatment was started at a stage of disease that would be otherwise fatal if left untreated. Antiviral treatment was associated with a rapid improvement in fever, ascites, lymphopenia and gross signs of illness and cats returned to normal health within 20 days or less of treatment. Significant reduction in viral titers was also observed in cats. These results indicate that continuous virus replication is required for progression of immune-mediated inflammatory disease of FIP. These findings may provide important insights into devising therapeutic strategies and selection of antiviral compounds for further development for important coronaviruses in animals and humans

Histologic discrepancy between endoscopic forceps biopsy and endoscopic mucosal resection specimens of colorectal polyp in actual clinical practice

Background/AimsWe aimed to assess the rate of histologic discrepancy (HD) between endoscopic forceps biopsy (EFB) and totally resected specimens in colorectal polyp and analyze the risk factors of discordant group, especially under-diagnosis (UD) cases before complete removal of colorectal polyp.MethodsFrom 2010 to 2015, a total of 290 polyps in 210 patients which had baseline pathology report before endoscopic resection (ER) were analyzed. UD cases were defined as those in which the diagnosis changed to a more advanced histologic feature after ER.ResultsA change in the final histology after ER was noted in 137 cases (47.2%), and after excluding 9 insignificant cases, 128 cases were further categorized into over-diagnosed and under-diagnosed group. UD occurred in 86 cases (29.7%) and change from benign to malignancy was noted in 26 cases (8.9%). On univariate analysis, a larger polyp size (>10 mm) was significantly associated with both HD (P10 mm was the single most significant predictor of both HD (P10 mm was the most important predictor of both HD and UD. We should be careful in making treatment strategy of colorectal polyp based on histologic report of EFB especially when the size of polyp is >10 mm

Transport and Fate of 137Cs Released From Multiple Sources in the North Atlantic and Arctic Oceans

The North Atlantic and Arctic oceans, along with the North Pacific, are the main reservoirs of anthropogenic radionuclides introduced in the past 75 years. The POSEIDON-R compartment model was applied to the North Atlantic and Arctic oceans to reconstruct 137Cs contamination in 1945–2020 due to multiple sources: global fallout, exchange flows with other oceans, point-source inputs in the ocean from reprocessing plants and other nuclear facilities, the impact of the Chernobyl accident and secondary contamination resulting from river runoff and redissolution from bottom sediments. The model simulated the marine environment as a system of 3D compartments comprising the water column, bottom sediment, and biota. The dynamic model described the transfer of 137Cs through the pelagic and benthic food chains. The simulation results were validated using the marine database MARIS. The calculated concentrations of 137Cs in the seaweed and non-piscivorous and piscivorous pelagic fish mostly followed the concentration of 137Cs in water. The concentration in coastal predator fish lagged behind the concentration in water as a result of a diet that includes both pelagic and benthic organisms. The impact of each considered source on the total concentration of 137Cs in non-piscivorous fish in the regions of interest was analyzed. Whereas the contribution from global fallout dominated in 1960–1970, in 1970–1990, the contribution of 137Cs released from reprocessing plants exceeded the contributions from other sources in almost all considered regions. Secondary contamination due to river runoff was less than 4% of ocean influx. The maximum total inventory of 137Cs in the Arctic Ocean (31,122 TBq) was reached in 1988, whereas the corresponding inventory in the bottom sediment was approximately 6% of the total. The general agreement between simulated and observed 137Cs concentrations in water and bottom sediment was confirmed by the estimates of geometric mean and geometric standard deviation, which varied from 0.89 to 1.29 and from 1.22 to 1.87, respectively. The approach used is useful to synthesize measurement and simulation data in areas with observational gaps. For this purpose, 13 representative regions in the North Atlantic and Arctic oceans were selected for monitoring by using the “etalon” method for classification