Understanding COVID-19 Situation in Nepal and Implications for SARS-CoV-2 Transmission and Management

CC BY-NC 4.0Emerging infectious diseases (EIDs) present one of the greatest challenges to public health in the 21st century. An emerging virus, depending on its potential to spread among humans, may cause individual or sporadic cases, culminating in a localized outbreak requiring public health intervention, or, in the worst-case scenario, a widespread epidemic, or worldwide pandemic.1 The novel Coronavirus Disease 2019 (COVID-19) is a new respiratory disease caused by Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) that is causing worldwide public health and economic challenges and has been recognized as a pandemic by the World Health Organization.2 The virus was first reported in Wuhan, Hubei Province, China, in December 2019.2,3 SARS-CoV-2 is an enveloped and positive single-stranded RNA virus belonging to the ß-coronavirus genus.2-4 SARS-CoV-2 holds high homology with SARS-CoV and targets angiotensin-converting enzyme receptor-2 (ACE2) for the viral attachment.4 A schematic depicting SARS-CoV-2 structure and pathogenesis is presented in Figure 1.5 There are very few studies on the transmission of SARS-CoV-2 through treated or untreated wastewater from advanced countries; however, COVID-19 surveillance of wastewater in developing countries has not been reported adequately.6 There is thus a need to study the presence of SARS-CoV-2 in wastewater in the developing countries like Nepal.OA publication support through Carolina Consortium agreement with Sag

Senecavirus A 3C Protease Mediates Host Cell Apoptosis Late in Infection

Senecavirus A (SVA), an oncolytic picornavirus used for cancer treatment in humans, has recently emerged as a vesicular disease (VD)-causing agent in swine worldwide. Notably, SVA-induced VD is indistinguishable from foot-and-mouth disease (FMD) and other high-consequence VDs of pigs. Here we investigated the role of apoptosis on infection and replication of SVA. Given the critical role of the nuclear factor-kappa B (NF-κB) signaling pathway on modulation of cell death, we first assessed activation of NF-κB during SVA infection. Results here show that while early during infection SVA induces activation of NF-κB, as evidenced by nuclear translocation of NF-κB-p65 and NF-κB-mediated transcription, late in infection a cleaved product corresponding to the C-terminus of NF-κB-p65 is detected in infected cells, resulting in lower NF-κB transcriptional activity. Additionally, we assessed the potential role of SVA 3C protease (3Cpro) in SVA-induced host-cell apoptosis and cleavage of NF-κB-p65. Transient expression of SVA 3Cpro was associated with cleavage of NF-κB-p65 and Poly (ADP-ribose) polymerase (PARP), suggesting its involvement in virus-induced apoptosis. Most importantly, we showed that while cleavage of NF-κB-p65 is secondary to caspase activation, the proteolytic activity of SVA 3Cpro is essential for induction of apoptosis. Experiments using the pan-caspase inhibitor Z-VAD-FMK confirmed the relevance of late apoptosis for SVA infection, indicating that SVA induces apoptosis, presumably, as a mechanism to facilitate virus release and/or spread from infected cells. Together, these results suggest an important role of apoptosis for SVA infection biology

Transcriptional Profiling of Human Brain Endothelial Cells Reveals Key Properties Crucial for Predictive In Vitro Blood-Brain Barrier Models

Brain microvascular endothelial cells (BEC) constitute the blood-brain barrier (BBB) which forms a dynamic interface between the blood and the central nervous system (CNS). This highly specialized interface restricts paracellular diffusion of fluids and solutes including chemicals, toxins and drugs from entering the brain. In this study we compared the transcriptome profiles of the human immortalized brain endothelial cell line hCMEC/D3 and human primary BEC. We identified transcriptional differences in immune response genes which are directly related to the immortalization procedure of the hCMEC/D3 cells. Interestingly, astrocytic co-culturing reduced cell adhesion and migration molecules in both BECs, which possibly could be related to regulation of immune surveillance of the CNS controlled by astrocytic cells within the neurovascular unit. By matching the transcriptome data from these two cell lines with published transcriptional data from freshly isolated mouse BECs, we discovered striking differences that could explain some of the limitations of using cultured BECs to study BBB properties. Key protein classes such as tight junction proteins, transporters and cell surface receptors show differing expression profiles. For example, the claudin-5, occludin and JAM2 expression is dramatically reduced in the two human BEC lines, which likely explains their low transcellular electric resistance and paracellular leakiness. In addition, the human BEC lines express low levels of unique brain endothelial transporters such as Glut1 and Pgp. Cell surface receptors such as LRP1, RAGE and the insulin receptor that are involved in receptor-mediated transport are also expressed at very low levels. Taken together, these data illustrate that BECs lose their unique protein expression pattern outside of their native environment and display a more generic endothelial cell phenotype. A collection of key genes that seems to be highly regulated by the local surroundings of BEC within the neurovascular unit are presented and discussed

Baseline characteristics of patients in the reduction of events with darbepoetin alfa in heart failure trial (RED-HF)

<p>Aims: This report describes the baseline characteristics of patients in the Reduction of Events with Darbepoetin alfa in Heart Failure trial (RED-HF) which is testing the hypothesis that anaemia correction with darbepoetin alfa will reduce the composite endpoint of death from any cause or hospital admission for worsening heart failure, and improve other outcomes.</p> <p>Methods and results: Key demographic, clinical, and laboratory findings, along with baseline treatment, are reported and compared with those of patients in other recent clinical trials in heart failure. Compared with other recent trials, RED-HF enrolled more elderly [mean age 70 (SD 11.4) years], female (41%), and black (9%) patients. RED-HF patients more often had diabetes (46%) and renal impairment (72% had an estimated glomerular filtration rate <60 mL/min/1.73 m2). Patients in RED-HF had heart failure of longer duration [5.3 (5.4) years], worse NYHA class (35% II, 63% III, and 2% IV), and more signs of congestion. Mean EF was 30% (6.8%). RED-HF patients were well treated at randomization, and pharmacological therapy at baseline was broadly similar to that of other recent trials, taking account of study-specific inclusion/exclusion criteria. Median (interquartile range) haemoglobin at baseline was 112 (106–117) g/L.</p> <p>Conclusion: The anaemic patients enrolled in RED-HF were older, moderately to markedly symptomatic, and had extensive co-morbidity.</p&gt

Improved functionalization of oleic acid-coated iron oxide nanoparticles for biomedical applications

Superparamagnetic iron oxide nanoparticles can providemultiple benefits for biomedical applications in aqueous environments such asmagnetic separation or magnetic resonance imaging. To increase the colloidal stability and allow subsequent reactions, the introduction of hydrophilic functional groups onto the particles’ surface is essential. During this process, the original coating is exchanged by preferably covalently bonded ligands such as trialkoxysilanes. The duration of the silane exchange reaction, which commonly takes more than 24 h, is an important drawback for this approach. In this paper, we present a novel method, which introduces ultrasonication as an energy source to dramatically accelerate this process, resulting in high-quality waterdispersible nanoparticles around 10 nmin size. To prove the generic character, different functional groups were introduced on the surface including polyethylene glycol chains, carboxylic acid, amine, and thiol groups. Their colloidal stability in various aqueous buffer solutions as well as human plasma and serum was investigated to allow implementation in biomedical and sensing applications.status: publishe

Antiinflammatory Therapy with Canakinumab for Atherosclerotic Disease

Background: Experimental and clinical data suggest that reducing inflammation without affecting lipid levels may reduce the risk of cardiovascular disease. Yet, the inflammatory hypothesis of atherothrombosis has remained unproved. Methods: We conducted a randomized, double-blind trial of canakinumab, a therapeutic monoclonal antibody targeting interleukin-1β, involving 10,061 patients with previous myocardial infarction and a high-sensitivity C-reactive protein level of 2 mg or more per liter. The trial compared three doses of canakinumab (50 mg, 150 mg, and 300 mg, administered subcutaneously every 3 months) with placebo. The primary efficacy end point was nonfatal myocardial infarction, nonfatal stroke, or cardiovascular death. RESULTS: At 48 months, the median reduction from baseline in the high-sensitivity C-reactive protein level was 26 percentage points greater in the group that received the 50-mg dose of canakinumab, 37 percentage points greater in the 150-mg group, and 41 percentage points greater in the 300-mg group than in the placebo group. Canakinumab did not reduce lipid levels from baseline. At a median follow-up of 3.7 years, the incidence rate for the primary end point was 4.50 events per 100 person-years in the placebo group, 4.11 events per 100 person-years in the 50-mg group, 3.86 events per 100 person-years in the 150-mg group, and 3.90 events per 100 person-years in the 300-mg group. The hazard ratios as compared with placebo were as follows: in the 50-mg group, 0.93 (95% confidence interval [CI], 0.80 to 1.07; P = 0.30); in the 150-mg group, 0.85 (95% CI, 0.74 to 0.98; P = 0.021); and in the 300-mg group, 0.86 (95% CI, 0.75 to 0.99; P = 0.031). The 150-mg dose, but not the other doses, met the prespecified multiplicity-adjusted threshold for statistical significance for the primary end point and the secondary end point that additionally included hospitalization for unstable angina that led to urgent revascularization (hazard ratio vs. placebo, 0.83; 95% CI, 0.73 to 0.95; P = 0.005). Canakinumab was associated with a higher incidence of fatal infection than was placebo. There was no significant difference in all-cause mortality (hazard ratio for all canakinumab doses vs. placebo, 0.94; 95% CI, 0.83 to 1.06; P = 0.31). Conclusions: Antiinflammatory therapy targeting the interleukin-1β innate immunity pathway with canakinumab at a dose of 150 mg every 3 months led to a significantly lower rate of recurrent cardiovascular events than placebo, independent of lipid-level lowering. (Funded by Novartis; CANTOS ClinicalTrials.gov number, NCT01327846.

Passive immunity to porcine epidemic diarrhea virusfollowing immunization of pregnant gilts with a recombinant orf virusvector expressing the spike protein

AbstractPassive immunity is critical for protection of neonatal piglets against porcine epidemic diarrhea virus (PEDV). Here, weinvestigated the immunogenicity of an orf virus (ORFV) vector expressing the full-length spike (S) protein of PEDV (ORFVPEDV-S) in pregnant gilts and its ability to confer passive immunity and protection in piglets. Three doses of ORFV-PEDV-Swere given to two groups of PEDV-negative pregnant gilts, with the last dose being administered two weeks prior to farrowing.One of the two groups immunized with the ORFV-PEDV-S recombinant virus was also exposed to live PEDV orallyon day 31 post-immunization (pi). Antibody responses were assessed in serum, colostrum and milk of immunized gilts, andpassive transfer of antibodies was evaluated in piglet sera. The protective efficacy of ORFV-PEDV-S was evaluated afterchallenge of the piglets with PEDV. PEDV-specific IgG, IgA and neutralizing antibody (NA) responses were detected inORFV-PEDV-S-immunized and ORFV-PEDV-S-immunized/PEDV-exposed gilts. PEDV NA, IgG and IgA were detected inthe serum of piglets born to immunized gilts, demonstrating the transfer of antibodies through colostrum and milk. Pigletsborn to immunized gilts showed reduced morbidity and a marked reduction in mortality after PEDV challenge in comparisonto control piglets. Piglets born to gilts that received ORFV-PEDV-S and were exposed to live PEDV showed stronger NAresponses and lower clinical scores when compared to piglets born to gilts immunized with ORFV-PEDV-S alone. Theseresults demonstrate the potential of ORFV as a vaccine delivery platform capable of eliciting passive immunity against PEDV

Understanding COVID-19 Situation in Nepal and Implications for SARS-CoV-2 Transmission and Management

Emerging infectious diseases (EIDs) present one of the greatest challenges to public health in the 21st century. An emerging virus, depending on its potential to spread among humans, may cause individual or sporadic cases, culminating in a localized outbreak requiring public health intervention, or, in the worst-case scenario, a widespread epidemic, or worldwide pandemic.1 The novel Coronavirus Disease 2019 (COVID-19) is a new respiratory disease caused by Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) that is causing worldwide public health and economic challenges and has been recognized as a pandemic by the World Health Organization.2 The virus was first reported in Wuhan, Hubei Province, China, in December 2019.2,3 SARS-CoV-2 is an enveloped and positive single-stranded RNA virus belonging to the ß-coronavirus genus.2-4 SARS-CoV-2 holds high homology with SARS-CoV and targets angiotensin-converting enzyme receptor-2 (ACE2) for the viral attachment.4 A schematic depicting SARS-CoV-2 structure and pathogenesis is presented in Figure 1.5 There are very few studies on the transmission of SARS-CoV-2 through treated or untreated wastewater from advanced countries\; however, COVID-19 surveillance of wastewater in developing countries has not been reported adequately.6 There is thus a need to study the presence of SARS-CoV-2 in wastewater in the developing countries like Nepal

Characterizing Nutritional, Antioxidant and Antimicrobial Values of Diploknema butyracea (Roxburgh) H. J. Lam from the Chepang Community, Makwanpur, Nepal

The multipurpose tree Diploknema butyracea (Roxb. H.J. Lam), known locally as Chiuri, is vital for food security and beekeeping in rural Nepal. This study examines its nutritional and phytochemical traits sourced from a Chepang community in Makwanpur, Nepal. This research focuses on macronutrients like carbohydrates, protein, fat, and ash alongside phytochemicals such as phenolic content, vitamin C, β-carotene, and lycopene. The study aimed to estimate this fruit's antimicrobial and antioxidant characteristics. The pulp and seed samples were analyzed for their nutritional and phytochemical components using standard methods (AOAC 1995). We determined the antioxidant and antimicrobial activity using the DPPH assay and agar diffusion method. This fruit has a high-fat content: 30.29% in the seed and 20.23% in the pulp. The pulp and seed also contain noteworthy levels of the total phenolic content (486.08 ± 0.006 and 182. 26 ± 0.001 mg Gallic Acid Equivalent (G.A.E.s) /100 g), vitamin C (20.70 ± 0.002 and 19.08 ± 0.005 mg Ascorbic Acid (A.A.)/100 g) with trace extents of compounds lycopene, β-carotene and carotenoids. We observed the antioxidant activity at 2207 ± 0.01 g/mL in pulp and 1841.05 ± 0.77 g/mL in seed, which is a substantial value. Both were discovered to be effective against Candida albicans at doses ranging from 25 to 100 mg/mL. By performing this study, we concluded that D. butyracea is a significant food source that can also be used medically