Search results for: viral pneumonia

Authors: Lian Zeng

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Double-Spear 1-H2-1 (DS1-H2-1) is an oncolytic virus and an innovative biological drug candidate. The chemical composition of the drug product is a live attenuated West Nile virus (WNV) containing the human T cell costimulator (CD86) gene. After intratumoral injection, the virus can rapidly self-replicate in the injected site and lyse/kill the tumor by repeated infection among tumor cells. We also established xenograft tumor models in mice to evaluate the drug candidate's efficacy on those tumors. The results from preclinical studies on transplanted tumors in immunodeficient mice showed that DS1-H2-1 had significant oncolytic effects on human-origin cancers: it completely (100%) shrieked human glioma; limited human neuroblastoma growth reached as high as 95% growth inhibition rate (%TGITW). The safety data of preclinical animal experiments confirmed that DS1-H2-1 is safe as a biological drug for clinical use. In the preclinical drug efficacy experiment, virus-drug administration with different doses did not show abnormal signs and disease symptoms in more than 300 tested mice, and no side effects or death occurred through various administration routes. Intravenous administration did not cause acute infectious disease or other side effects. However, the replication capacity of the virus in tumor tissue via intravenous administration is only 1% of that of direct intratumoral administration. The direct intratumoral administration of DS1-H2-1 had a higher rate of viral replication. Therefore, choosing direct intratumoral injection can ensure both efficacy and safety.

Keywords: oncolytic virus, WNV-CD86, immunotherapy drugs, glioma, neuroblastoma

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Authors: I. Schäfer, B. Kohn, M. Volkmann, E. Müller

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Introduction: Blood-feeding arthropods transmit parasitic, bacterial, or viral pathogens to domestic animals and wildlife. Vector-borne infections are gaining significance due to the increase of travel, import of domestic animals from abroad, and the changing climate in Europe. Aims of the study: The main objective of this retrospective study was to assess the prevalence of vector-borne infections in cats in which a ‘Feline Travel Profile’ had been conducted. Material and Methods: This retrospective study included test results from cats for which a ‘Feline Travel Profile’ established by LABOKLIN had been requested by veterinarians between April 2012 and December 2019. This profile contains direct detection methods via polymerase chain reaction (PCR) for Hepatozoon spp. and Dirofilaria spp. as well as indirect detection methods via immunofluorescence antibody test (IFAT) for Ehrlichia spp. and Leishmania spp. This profile was expanded to include an IFAT for Rickettsia spp. from July 2015 onwards. The prevalence of the different vector-borne infectious agents was calculated. Results: A total of 602 cats were tested using the ‘Feline Travel Profile’. Positive test results were as follows: Rickettsia spp. IFAT 54/442 (12.2%), Ehrlichia spp. IFAT 68/602 (11.3%), Leishmania spp. IFAT 21/602 (3.5%), Hepatozoon spp. PCR 51/595 (8.6%), and Dirofilaria spp. PCR 1/595 cats (0.2%). Co-infections with more than one pathogen could be detected in 22/602 cats. Conclusions: 170/602 cats (28.2%) were tested positive for at least one vector-borne pathogen. Infections with multiple pathogens could be detected in 3.7% of the cats. The data emphasizes the importance of considering vector-borne infections as potential differential diagnoses in cats.

Keywords: arthopod-transmitted infections, feline vector-borne infections, Germany, laboratory diagnostics

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Authors: Fumio Kasai, Noriko Hirayama, Jorge Pereira, Azusa Ohtani, Masashi Iemura, Malcolm A. Ferguson Smith, Arihiro Kohara

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The VERO cell line was established in 1962 from normal tissue of an African green monkey, Chlorocebus aethiops (2n=60), and has been commonly used worldwide for screening for toxins or as a cell substrate for the production of viral vaccines. The VERO genome was sequenced in 2014; however, its cytogenetic features have not been fully characterized as it contains several chromosome abnormalities and different karyotypes coexist in the cell line. In this study, the VERO cell line (JCRB0111) was compared with one of the sublines. In contrast to 59 chromosomes as the modal chromosome number in the VERO cell line, the subline had two peaks of 56 and 58 chromosomes. M-FISH analysis using human probes revealed that the VERO cell line was characterized by a translocation t(2;25) found in all metaphases, which was absent in the subline. Different abnormalities detected only in the subline show that the cell line is heterogeneous, indicating that the subline has the potential to change its genomic characteristics during cell culture. The various alterations in the two independent lineages suggest that genomic changes in both VERO cells can be accounted for by progressive rearrangements during their evolution in culture. Both t(5;X) and t(8;14) observed in all metaphases of the two cell lines might have a key role in VERO cells and could be used as genetic markers to identify VERO cells. The flow karyotype shows distinct differences from normal. Further analysis of sorted abnormal chromosomes may uncover other characteristics of VERO cells. Because of the absence of STR data, cytogenetic data are important in characterizing animal cell lines and can be an indicator of their quality control.

Keywords: VERO, cell culture passage, chromosome rearrangement, heterogeneous cells

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Authors: Sweta Pandey, Samridhi Dhyani, Susmita Chaudhuri

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Klebsiella pneumoniae causes a wide range of infections, including urinary tract infections, sepsis, bacteremia, pneumonia, and liver abscesses. The emergence of multi-drug resistance in this bacterium led to a major setback for clinical management. WHO also endorsed a need for finding alternative therapy to antibiotics for the treatment of these infections. Development of vaccines and passive antibody therapy has been proven as a potent alternative to antibiotics in the case of MDR, XDR, and PDR Klebsiella infections. Siderophore receptors have been demonstrated to be overexpressed for the internalization of iron siderophore complexes during infections in most Gram-negative bacteria. For the present study, immune response to siderophore receptors to establish this protein as a potential immunogen for the development of therapeutic leads was explored. Clinical strains of Klebsiella pneumoniae were grown in iron-deficient conditions, and the iron-regulated outer membrane proteins were extracted and characterized through mass spectrometry for specific identification. The gene for identified protein was cloned in pET- 28a vector and expressed in E. coli. The native protein and the recombinant protein were isolated and purified and used as antigens for the generation of immune response in BALB/c mice. The native protein of Klebsiella pneumoniae grown in iron-deficient conditions was identified as FepA (Ferrienterobactin receptor) and other siderophore receptors. This 80 kDa protein generated an immune response in BALB/c mice. The antiserum from mice after subsequent booster doses was collected and showed binding with FepA protein in western blot and phagocytic uptake of the K. pneumoniae in the presence antiserum from immunized mice also observed from the animal studies after bacterial challenge post immunisation in mice have shown bacterial clearance. The antiserum from mice showed binding and clearance of the Klebsiella pneumoniae bacteria in vitro and in vivo. These antigens used for generating an active immune response in mice can further be used for therapeutic monoclonal antibody development against Klebsiella pneumoniae infections.

Keywords: antiserum, FepA, Klebsiella pneumoniae, multi drug resistance, siderophore receptor

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Authors: Gyati Shilakari Asthana, Abhay Asthana, Dharm Veer Kohli, Suresh Prasad Vyas

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Purpose: The therapeutic potential of oligonucleotide (ODN) is primarily dependent upon its safe and efficient delivery to specific cells overcoming degradation and maximizing cellular uptake in vivo. The present study is focused to design low molecular weight chitosan nanoconstructs to meet the requirements of safe and effectual delivery of ODNs. LMW-chitosan is a biodegradable, water soluble, biocompatible polymer and is useful as a non-viral vector for gene delivery due to its better stability in water. Methods: LMW chitosan ODN nanoparticles (CHODN NPs) were formulated by self-assembled method using various N/P ratios (moles ratio of amine groups of CH to phosphate moieties of ODNs; 0.5:1, 1:1, 3:1, 5:1, and 7:1) of CH to ODN. The developed CHODN NPs were evaluated with respect to gel retardation assay, particle size, zeta potential and cytotoxicity and transfection efficiency. Results: Complete complexation of CH/ODN was achieved at the charge ratio of 0.5:1 or above and CHODN NPs displayed resistance against DNase I. On increasing the N/P ratio of CH/ODN, the particle size of the NPs decreased whereas zeta potential (ZV) value increased. No significant toxicity was observed at all CH concentrations. The transfection efficiency was increased on increasing N/P ratio from 1:1 to 3:1, whereas it was decreased with further increment in N/P ratio upto 7:1. Maximum transfection of CHODN NPs with both the cell lines (Raw 267.4 cells and Hela cells) was achieved at N/P ratio of 3:1. The results suggest that transfection efficiency of CHODN NPs is dependent on N/P ratio. Conclusion: Thus the present study states that LMW chitosan nanoparticulate carriers would be acceptable choice to improve transfection efficiency in vitro as well as in vivo delivery of oligonucleotide.

Keywords: LMW-chitosan, chitosan nanoparticles, biocompatibility, cytotoxicity study, transfection efficiency, oligonucleotide

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Authors: Nickolay Nosik, Dmitry Nosik, Marina Bochkova, Nina Kondrashina, Olga Lobach

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The bactericidal effect of UV radiation is known for long time and widely used for inactivation of pathogens but for viruses it is not so uniform. Due to a wide variety of viruses their sensitivity to UV radiation is quite different and not quite predictable. The goal of the study was to determine the inactivation kinetics of UV radiation ( 254 nm) of the viruses of social importance (HIV), as well as test-viruses (poliovirus, adenovirus) used for the evaluation of the viral inactivation efficacy of germicides. Methods: DNA viruses- adenovirus, type 5; Herpes simplex virus (HSV), type 1, and RNA viruses–human immunodeficiency virus (HIV), type 1 and poliovirus, type 1 (Sabin strain) were obtained from State collection of viruses ( The D.I. Ivanovsky Institute of Virology). The source of UV radiation was a 15-watt low-pressure mercury vapor lamp (over 60% 254nm). The samples of 5cm2 were placed direct under the UV lamp flow (h-0.3m). Log reduction value was used as a marker for the rate of virus inactivation. Results: The data obtained indicate that poliovirus (one of the viruses most resistant to chemical germicides) and HSV are rather sensitive to UV radiation ( D90 =250-311 J/m2). Adenovirus is much more resistant to UV radiation (750 J/m2 ). The kinetics of adenovirus inactivation : 0 min- 5.0 lg TCID50, 10 min - 5,0, 15 min -4,0, 30 min – 3.5, 60 min – 1,0, 75 min -0,5 lg TCID50, 90 min –virus not detectable. HIV is most resistant to UV radiation among the studied viruses. It takes more than 4 hrs to inactivate the virus on the surface. D90 = 2000 J/m2 Conclusion: The results of the study show that there is no direct dependence between sensitivity to UV light and the size of the virion or presence\absence of the envelope of the virus. Poliovirus and adenovirus are small viruses (20-30nm poliovirus and 70-90nm adenovirus) and both are non-enveloped viruses but adenovirus 3-fold more resistant to UV radiation than poliovirus. It can be expected that viruses with more complicate structure, like Herpes virus (200nm) or HIV (80-100 nm), would be more sensitive to UV light. However, the very high resistance of HIV to UV radiation needs further investigation. The diverse resistance of the different viruses to UV radiation should be taken into the account when UV light is used to inactivate infectious viruses in hospitals and other public environments.

Keywords: HIV, HSV, inhibition of viruses, UV radiation

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Authors: Doreen Duri, Danai Zhou, Babil Stray-Pedersen, Collet Dandara

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Given the high prevalence of HIV/AIDS in sub-Saharan Africa, and the elusive search for a cure, understanding the pharmacogenetics of currently used drugs is critical in populations from the most affected regions. Compared to Asian and Caucasian populations, African population groups are more genetically diverse, making it difficult to extrapolate findings from one ethnic group to another. This study aimed to investigate the role of genetic variation in CYP2B6 (c.516G>T and c.983T>C) single nucleotide polymorphisms on plasma nevirapine levels among HIV-infected adult Zimbabwean patients. Using a cross-sectional study, patients on nevirapine-containing HAART, having reached steady state (more than six weeks on treatment) were recruited to participate. Blood samples were collected after patients provided consent and samples were used to extract DNA for genetic analysis or to measure plasma nevirapine levels. Genetic analysis was carried out using PCR and RFLP or Snapshot for the two single nucleotide polymorphisms; CYP2B6 c.516G>T and c.983T>C, while LC-MS/MS was used in analyzing nevirapine concentration. CYP2B6 c.516G>T and c.983T>C significantly predicted plasma nevirapine concentration with the c.516T and c.983T being associated with elevated plasma nevirapine concentrations. Comparisons of the variant allele frequencies observed in this group to those reported in some African, Caucasian and Asian populations showed significant differences. We conclude that pharmacogenetics of nevirapine can be creatively used to determine patients who are likely to develop nevirapine-associated side effects as well as too low plasma concentrations for viral suppression.

Keywords: allele frequencies, genetically diverse, nevirapine, single nucleotide polymorphism

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Authors: Constant Gillot, Pauline Michaux, Julien Favresse, Jean-Michel Dogné, Jonathan Douxfils

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Introduction: NETosis has emerged as a crucial yet paradoxical factor in severe COVID-19 cases. While neutrophil extracellular traps (NETs) help contain and eliminate viral particles, excessive NET formation can lead to hyperinflammation, exacerbating tissue damage and acute respiratory distress syndrome (ARDS). Aims: This study evaluates the relationship between COVID-19-infected sera and NETosis using an ex-vivo model. Methods: Sera from 8 post-admission COVID-19 patients, after receiving corticoid therapy, were used to induce NETosis in neutrophils from a healthy donor. NET formation was tracked using fluorescent markers for DNA and neutrophil elastase (NE) every 2 minutes for 8 hours. The results were expressed as a percentage of DNA/NE released over time. Key metrics, including T50 (time to 50% release) and AUC (area under the curve), representing total NETosis potential), were calculated. A 27-cytokine screening kit was used to assess the cytokine composition of the sera. Results: COVID-19 sera induced NETosis based on their cytokine profile. The AUC of NE and DNA release decreased with time following corticoid therapy, showing a significant reduction in 6 of the 8 patients (p<0.05). T50 also decreased in parallel with AUC for both markers. Cytokines concentration decrease with time after therapy administration. There is correlation between 14 cytokines concentration and NE release. Conclusion: This ex-vivo model successfully demonstrated the induction of NETosis by COVID-19 sera using two markers. A clear decrease in NETosis potential was observed over time with glucocorticoid therapy. This model can be a valuable tool for monitoring NETosis and investigating potential NETosis inducers and inhibitors.

Keywords: NETosis, COVID-19, cytokine storm, biomarkers

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Authors: Gwendy Darras, Mattias Desmet

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Previous research showed that psychological distress has a negative impact on the disease course of viral infections. For COVID-19, the same association was observed in small samples of specific segments of the population (e.g. health care workers). The present study presents a more refined analysis of this association, measuring a broader spectrum of psychological distress in a large sample (n=3084) of the general Flemish population. Several types of psychological distress (state, trait and health anxiety, depression, intra-, and interpersonal stress) are registered throughout three periods: one year before the contamination, one week before the contamination, and during the contamination. In doing so, validated scales such as DASS-21, IIP-32, and FCV-19S are used. Furthermore, the course of COVID-19 is registered in several ways: number of symptoms, number of days sick leave due to COVID-19, and number of days the symptoms have lasted. Also, different control variables such as vaccination status, medical and psychological history are taken into account. Statistical analysis shows that all types of psychological distress are positively correlated with the severity of the COVID-19 disease course. Anxiety during the contamination shows the strongest correlation, but psychological distress one year before the onset of COVID-19 was still significantly associated with the worsening of the disease course. As the assessment of the latter type of distress happened before the onset of the COVID-19 disease course, retrospective bias resulting in artificial associations between self-reported stress and COVID-19 severity is unlikely to have impacted the observations. In view of possible future pandemics, it is important to focus on general stress and anxiety reduction in the general population as soon as possible. It is also advisable to minimize the use of stress-inducing messages to encourage the population to adhere to the measures issued during a pandemic.

Keywords: anxiety, COVID-19, depression, psychoneuroimmunology, psychological distress, stress

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Authors: Driss Cherqaoui, Nouhaila Ait Lahcen, Ismail Hdoufane, Mehdi Oubahmane, Wissal Liman, Christelle Delaite, Mohammed M. Alanazi

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The Ebola virus is a highly contagious and deadly pathogen that causes Ebola virus disease. The Ebola virus glycoprotein (EBOV-GP) is a key factor in viral entry into host cells, making it a critical target for therapeutic intervention. Using a combination of computational approaches, this study focuses on the identification of natural compounds that could serve as potent inhibitors of EBOV-GP. The 3D structure of EBOV-GP was selected, with missing residues modeled, and this structure was minimized and equilibrated. Two large natural compound databases, COCONUT and NPASS, were chosen and filtered based on toxicity risks and Lipinski’s Rule of Five to ensure drug-likeness. Following this, a pharmacophore model, built from 22 reported active inhibitors, was employed to refine the selection of compounds with a focus on structural relevance to known Ebola inhibitors. The filtered compounds were subjected to virtual screening via molecular docking, which identified ten promising candidates (five from each database) with strong binding affinities to EBOV-GP. These compounds were then validated through molecular dynamics simulations to evaluate their binding stability and interactions with the target. The top three compounds from each database were further analyzed using ADMET profiling, confirming their favorable pharmacokinetic properties, stability, and safety. These results suggest that the selected compounds have the potential to inhibit EBOV-GP, offering new avenues for antiviral drug development against the Ebola virus.

Keywords: EBOV-GP, Ebola virus glycoprotein, high-throughput drug screening, molecular docking, molecular dynamics, natural compounds, pharmacophore modeling, virtual screening

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Authors: Alieh Farshbaf

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Introduction: Current anti-retroviral drugs have some restrictions for treatment of HIV-1 infection. The efficacy of retroviral drugs is not same in different infected patients and the virus rebound from latent reservoirs after stopping them. Recently, the engineering of stem cells and gene therapy provide new approaches to eliminate some drug problems by induction of resistance to HIV-1. Literature review: Up to now, AIDS-restriction genes (ARGs) were suitable candidate for gene and cell therapies, such as cc-chemokine receptor-5 (CCR5). In this manner, CCR5 provide effective cure in Berlin and Boston patients by inducing of HIV-1 resistance with allogeneic stem cell transplantation. It is showed that Zinc Finger Nuclease (ZFN) could induce HIV-1 resistance in stem cells of infected patients by homologous recombination or non-end joining mechanism and eliminate virus loading after returning the modified cells. Then, gene modification by HIV restriction factors, as TRIM5, introduced another gene candidate for HIV by interfering in infection process. These gene modifications/editing provided by stem cell futures that improve treatment in refractory disease such as HIV-1. Conclusion: Although stem cell transplantation has some complications, but in compare to retro-viral drugs demonstrated effective cure by elimination of virus loading. On the other hand, gene therapy is cost-effective for an infected patient than retroviral drugs payment in a person life-long. The results of umbilical cord blood stem cell transplantation showed that gene and cell therapy will be applied easier than previous treatment of AIDS with high efficacy.

Keywords: stem cell, AIDS, gene modification, cell engineering

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Authors: Pampita Chakraborty, Sukumar Mukherjee

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Diabetes Mellitus (DM) is commonly encountered metabolic disorder in clinical practice. An estimated 25 percent of DM patients develop foot problems. Foot ulceration and infection are one of the major causes of morbidity, hospitalization or even amputation. Objective: To isolate and identify bacterial pathogens in Diabetic Foot Ulcer (DFU) and to observe its antimicrobial sensitivity pattern. Methodology: A prospective study was conducted for a period of 9 months at the Department of Microbiology, GD Hospital & Diabetes Institute, Kolkata. 75 DFU patients were recruited in the study. Specimens for microbiological studies obtained from ulcer base were examined as gram stained smear and was cultured aerobically on Nutrient agar, Blood agar and MacConkey agar plates. Antimicrobial sensitivity test was performed by disc diffusion techniques according to CLSI guidelines. Result: In this study out of 75cases, 73% (55/75) were male and 27% (20/75) were females with mean (SD) age of 51.11(±10) years. Out of 75 pus cultures, 63(84%) showed growth of microorganism making total of 81 bacterial isolates with 71.42% of monomicrobial infection and 28.57% of polymicrobial infection. Out of 81 isolates 53(65.43%) were gram negative and 21(25.92%) were gram positive. E.coli was relatively common isolate 21(26%) followed by Staphylococcus aureus 15(18.5%), Klebsiella pneumonia 14(17.28%), Pseudomonas aeruginosa 12 (14.81%), Proteus spp. 3 (3.70%), and Enterococcus faecalis 6 (7.40%). 75% of Gram-negative microorganism were extended Beta-lactamase enzyme (ESBL) producer and around 20 % of Klebsiella and Proteus spp. were carbapenemase enzyme producer. Among Gram positive, around 50% of S.aureus was MRSA, sensitive only to Vancomycin, Teicoplanin & Linezolid. Conclusion: More prevalence of monomicrobial gram-negative bacteria than gram-positive bacteria in DFU was observed. This study emphasizes that Beta-Lactam group of antibiotics should not be the empirical treatment of choice for Gram-negative isolates; instead alternatives like Carbapenems, Amikacin could be a better option. On the other hand, Vancomycin and Linezolid are preferred for most of the infection with gram-positive aerobes. Continuous surveillance of resistant bacteria is required for empiric therapy.

Keywords: antibiotic resistant, antimicrobial susceptibility, diabetic foot ulcer, surveillance

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Authors: Ali Faisal Saleem, Farheen Quadri, Mach Ondrej, Anita Zaidi

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Purpose: Pakistan is the final frontier for a polio-free world. Chronic malnutrition is associated with lack of effective gut immunity, and possibly associated with poliomyelitis in children received multiple OPV. We evaluate IPV dose administered together with OPV results in higher immunogenicity and mucosal immunity compared to OPV alone in chronically malnourished infants. Methods AND Materials: A community-based, unblinded-randomized-trial, conducted in 5 peri-urban, low-middle-income households of Karachi, in infants 9-12 months. Two study groups were non-malnourished (HAZ= -2 or more) and chronic malnourished (HAZ <-2SD), with 2-arms each i) OPV and ii) OPV and IPV. Two blood specimens (2ml) at baseline and at day 28 and two stool specimens (6 gm.) at day 29 and after 7 days. All infants received a bOPV challenge dose after first stool specimen. Calculates sample size was 210 in each arm. Serological (baseline compared to 28 days post-vaccine) and mucosal immunity after one week of bOPV challenge dose were study outcomes. Results: Baseline seroprevalence in malnourished infants were low compared to non-malnourished (P1, P2 and P3 (p=<0.001). There is significant rise in antibody titer and P1 seroprevalence in Mal A and B after receiving study vaccine; much higher in Mal B. Infants randomized to bOPV + IPV study vaccine showed incremental immune response against P1 (Mal B, 92.2%; Nor B, 98.4%), P2 (Mal B, 90.4%; Nor B, 94.7%), and P3 (Mal B, 85.6% and Nor B, 93.5%) was observed. A significant proportion of infants in malnourished (P1, 13%; P2, 24%; P3, 26%) and normally nourished group (P1, 5%; P2, 11%; P3, 14%) were found to be seronegative at baseline. Infants who received BOPV + IPV as their study vaccine showed a very high seroconversion response after vaccine (p=<0.001 for P1, P2 and P3). Majority of the specimens were negative at baseline (Mal A, 2%, Mal B, 1%; Nor A, 2%; Nor B, 1%), and remains negative after bOPV challenge dose (Mal A, 8%, Mal B, 6%; Nor A, 11%; Nor B, 10%). Conclusion: Malnourished-infants have low poliovirus-seroprevalence that increased remarkably after IPV. There is less viral shedding after IPV in infants.

Keywords: chronic malnutrition, infants, IPV, OPV

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Authors: Daria Reczynska, Justyna Jarczak, Michal Czopowicz, Danuta Sloniewska, Karina Horbanczuk, Wieslaw Jarmuz, Jaroslaw Kaba, Emilia Bagnicka

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Since people, animals and plants are constantly exposed to pathogens they have developed very complex systems of defense. Among ca. 1000 antimicrobial peptides from different families so far identified, approximately 30 belonging to cathelicidin family can be found in mammals. Cathelicidins probably constitute the first line of defense because they can act at a physiological salt concentration which is present in healthy tissues. Moreover, the low salt concentration which is present in infected tissues inhibits their activity. In goat bactenecin 7.5 (BAC7.5), bactenecin 5 (BAC5), myeloid antimicrobial peptide 28 (MAP28), myeloid antimicrobial peptide 34 (MAP34 A and B), goat bactenecin3.4 (ChBac3.4) were identified. Caprine arthritis-encephalitis (CAE) caused by small ruminant lentivirus (SRLV) is economic problem. The main CAE symptoms are weight loss, arthritis, pneumonia and mastitis (significant elevation of the somatic cell count and deterioration of some technological parameters). The study was conducted on 24 dairy goats. The animals were divided into two groups: experimental (SRLV-infected) and control (non-infected). The blood samples were collected five times: on the 1st, 7th, 30th, 90th and 150thday of lactation. The levels of transcripts of BAC7.5, BAC5, MAP28 and MAP34 genes in blood leucocytes were measured using qPCR method. There were no differences in mRNA levels of studied genes between stages of lactation. The differences were observed in expressions of BAC5, MAP28 and MAP34 genes with lower levels in the experimental group. There was no difference in BAC7.5 expression between groups. The decreased levels of transcripts of cathelicidin genes in blood leucocytes of SRLV-infected goats may indicate the disturbances of homeostasis in organisms. It can be concluded that SRLV infection seems to inhibit expression of cathelicidin genes. The study was financed by a grant from the National Scientific Center No. UMO-2013/09/B/NZ/03514.

Keywords: goat, CAEV, cathelicidins, blood leukocytes, gene expression

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Authors: Maria L. G. Lourenço, Keylla H. N. P. Pereira, Viviane Y. Hibaru, Fabiana F. Souza, João C. P. Ferreira, Simone B. Chiacchio, Luiz H. A. Machado

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Orphaned is a risk factor for mortality in newborns since it is a condition with total or partial absence of maternal care that is essential for neonatal survival, including nursing (nutrition, the transference of passive immunity and hydration), warmth, urination, and defecation stimuli, and protection. The most common causes of mortality in orphans are related to lack of assistance, handling mistakes and infections. This study aims to describe the orphans rates in neonatal puppies, the main causes, and the mortality rates. The study included 735 neonates admitted to the Sao Paulo State University (UNESP) Veterinary Hospital, Botucatu, Sao Paulo, Brazil, between January 2018 and November 2019. The orphans rate was 43.4% (319/735) of all neonates included, and the main causes for orphaned were related to maternal agalactia/hypogalactia (23.5%, 75/319); numerous litter (15.7%, 50/319), toxic milk syndrome due to maternal mastitis (14.4%, 46/319), absence of suction/weak neonate (12.2%, 39/319), maternal disease (9.4%, 30/319), cleft palate/lip (6.3%, 20/319), maternal death (5.9%, 19/319), prematurity (5.3%, 17/319), rejection/failure in maternal instinct (3.8%, 12/319) and abandonment by the owner/separation of mother and neonate (3.5%, 11/319). The main consequences of orphaned observed in the admitted neonates were hypoglycemia, hypothermia, dehydration, aspiration pneumonia, wasting syndrome, failure in the transference of passive immunity, infections and sepsis, which happened due to failure of identifying the problem early, lack of adequate assistance, negligence and handling mistakes by the owner. The total neonatal mortality rate was 8% (59/735) and the neonatal mortality rate among orphans was 18.5% (59/319). The orphaned and mortality rates were considered high, but even higher rates may be observed in locations without adequate neonatal assistance and owner orientation. The survival of these patients is related to constant monitoring of the litter, early diagnosis and assistance, and the implementation of effective handling for orphans. Understanding the correct handling for neonates and instructing the owners regarding proper handling are essential to minimize the consequences of orphaned and the mortality rates.

Keywords: orphans, neonatal care, puppies, newborn dogs

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Authors: Chan Eang Teng, Tang Mui Joo

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With the viral factor on social media, the sense of persuasion is generated by repetition and popularity. When users’ interest is captured, political awareness increases to spark political enthusiasm, but, the level of user’s political participation and political attitude of those active users is still questionable. An online survey on 250 youth and in-depth interview on two politicians are conducted to answer the main question in this paper. The result shows that Facebook significantly increases political awareness among youths. Social media may not be the major trigger to political activism among youths as most respondents opined that they would still vote without Facebook. Other factors could be political campaigning, political climate, age, peer pressure or others. Finding also shows that majority of respondents did not participate in online political debates or political groups. Many also wondered if the social media was the main power switch that triggers the political influx among young voters. The research finding is significant to understand how the new media, Facebook, has reshaped the political landscape in Malaysia, creating the Social Media Election that changed the rules of the political game. However, research finding does not support the ideal notion that the social media is the major trigger to youth’s political activism. This research outcome has exposed the flaws of the Social Media Election. It has revealed the less optimistic side of youth political activism. Unfortunately, results fall short of the idealistic belief that the social media have given rise to political activism among youths in the 13^th General Election in Malaysia. The research outcome also highlights an important lesson for the democratic discourse of Malaysia which is making informed and educated decisions takes more commitment, proactive and objective attitude.

Keywords: social media, political participation, political activism, democracy, political communication

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Authors: Yi-Lin Chen, Sheng-Jou Hung, Tsunglin Liu

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Adaptive immune system recognizes a wide range of antigens via expressing a large number of structurally distinct T cell and B cell receptor genes. The distinct receptor genes arise from complex rearrangements called V(D)J recombination, and constitute the immune repertoire. A common method of profiling immune repertoire is via amplifying recombined receptor genes using multiple primers and high-throughput sequencing. This multiplex-PCR approach is efficient; however, the resulting repertoire can be distorted because of primer bias. To eliminate primer bias, 5’ RACE is an alternative amplification approach. However, the application of RACE approach is limited by its low efficiency (i.e., the majority of data are non-regular receptor sequences, e.g., containing intronic segments) and lack of the convenient tool for analysis. We propose a computational tool that can correctly identify non-regular receptor sequences in RACE data via aligning receptor sequences against the whole gene instead of only the exon regions as done in all other tools. Using our tool, the remaining regular data allow for an accurate profiling of immune repertoire. In addition, a RACE approach is improved to yield a higher fraction of regular T-cell receptor sequences. Finally, we quantify the degree of primer bias of a multiplex-PCR approach via comparing it to the RACE approach. The results reveal significant differences in frequency of VJ combination by the two approaches. Together, we provide a new experimental and computation pipeline for an unbiased profiling of immune repertoire. As immune repertoire profiling has many applications, e.g., tracing bacterial and viral infection, detection of T cell lymphoma and minimal residual disease, monitoring cancer immunotherapy, etc., our work should benefit scientists who are interested in the applications.

Keywords: immune repertoire, T-cell receptor, 5' RACE, high-throughput sequencing, sequence alignment

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Authors: Cornelius Arome Omatola, Ropo Ebenezer Ogunsakin, Anyebe Bernard Onoja, Martin-Luther Oseni Okolo, Joseph Abraham-Oyiguh, Kehinde Charles Mofolorunso, Phoebe Queen Akoh, Omebije Patience Adejo, Joshua Idakwo, Therisa Ojomideju Okeme, Danjuma Muhammed, David Moses Adaji, Sunday Ocholi Samson, Ruth Aminu, Monday Eneojo Akor

Abstract:

Gastroenteritis viruses are the leading etiologic agents of diarrhea in children worldwide. We present data from thirty-three (33) eligible studies published between 2003 and 2023 from African countries bearing the brunt of the virus-associated diarrheal mortality. Random effects meta-analysis with proportion, subgroups, and meta-regression analyses were employed. Overall, rotavirus with estimated pooled prevalence of 31.0% (95% CI 24.0–39.0) predominated in all primary care visits and hospitalizations, followed by norovirus, adenovirus, sapovirus, astrovirus, and aichivirus with pooled prevalence estimated at 15.0% (95% CI 12.0–20.0), 10% (95% CI 6-15), 4.0% (95% CI 2.0–6.0), 4% (95% CI 3-6), and 2.3% (95% CI 1-3), respectively. Predominant rotavirus genotype was G1P[8] (38%), followed by G3P[8] (11.7%), G9P[8] (8.7%), and G2P[4] (7.1%); although, unusual genotypes were also observed, including G3P[6] (2.7%), G8P[6] (1.7%), G1P[6] (1.5%), G10P[8] (0.9%), G8P[4] (0.5%), and G4P[8] (0.4%). The genogroup II norovirus predominated over the genogroup I-associated infections (84.6%, 613/725 vs 14.9%, 108/725), with the GII.4 (79.3%) being the most prevalent circulating genotype. In conclusion, this review showed that rotavirus remains the leading driver of viral diarrhea requiring health care visits and hospitalization among under-five years children in Africa. Thus, improved rotavirus vaccination in the region and surveillance to determine the residual burden of rotavirus and the evolving trend of other enteric viruses are needed for effective control and management of cases.

Keywords: enteric viruses, rotavirus, norovirus, adenovirus, astrovirus, gastroenteritis

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Authors: S. Ben Fredj, H. Ghali, M. Ben Rejeb, S. Layouni, S. Khefacha, L. Dhidah, H. Said

Abstract:

Background: The Intensive Care Unit (ICU) provides continuous care and uses a high level of treatment technologies. Although developed country hospitals allocate only 5–10% of beds in critical care areas, approximately 20% of nosocomial infections (NI) occur among patients treated in ICUs. Whereas in the developing countries the situation is still less accurate. The aim of our study is to assess mortality rates in ICUs and to determine its predictive factors. Methods: We carried out a nested case-control study in a 630-beds public tertiary care hospital in Eastern Tunisia. We included in the study all patients hospitalized for more than two days in the surgical or medical ICU during the entire period of the surveillance. Cases were patients who died before ICU discharge, whereas controls were patients who survived to discharge. NIs were diagnosed according to the definitions of ‘Comité Technique des Infections Nosocomiales et les Infections Liées aux Soins’ (CTINLIS, France). Data collection was based on the protocol of Rea-RAISIN 2009 of the National Institute for Health Watch (InVS, France). Results: Overall, 301 patients were enrolled from medical and surgical ICUs. The mean age was 44.8 ± 21.3 years. The crude ICU mortality rate was 20.6% (62/301). It was 35.8% for patients who acquired at least one NI during their stay in ICU and 16.2% for those without any NI, yielding an overall crude excess mortality rate of 19.6% (OR= 2.9, 95% CI, 1.6 to 5.3). The population-attributable fraction due to ICU-NI in patients who died before ICU discharge was 23.46% (95% CI, 13.43%–29.04%). Overall, 62 case-patients were compared to 239 control patients for the final analysis. Case patients and control patients differed by age (p=0,003), simplified acute physiology score II (p < 10-3), NI (p < 10-3), nosocomial pneumonia (p=0.008), infection upon admission (p=0.002), immunosuppression (p=0.006), days of intubation (p < 10-3), tracheostomy (p=0.004), days with urinary catheterization (p < 10-3), days with CVC ( p=0.03), and length of stay in ICU (p=0.003). Multivariate analysis demonstrated 3 factors: age older than 65 years (OR, 5.78 [95% CI, 2.03-16.05] p=0.001), duration of intubation 1-10 days (OR, 6.82 [95% CI, [1.90-24.45] p=0.003), duration of intubation > 10 days (OR, 11.11 [95% CI, [2.85-43.28] p=0.001), duration of CVC 1-7 days (OR, 6.85[95% CI, [1.71-27.45] p=0.007) and duration of CVC > 7 days (OR, 5.55[95% CI, [1.70-18.04] p=0.004). Conclusion: While surveillance provides important baseline data, successful trials with more active intervention protocols, adopting multimodal approach for the prevention of nosocomial infection incited us to think about the feasibility of similar trial in our context. Therefore, the implementation of an efficient infection control strategy is a crucial step to improve the quality of care.

Keywords: intensive care unit, mortality, nosocomial infection, risk factors

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Authors: O. Antypenko, I. Vasilieva, S. Kovalenko

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Investigations for new effective and less toxic antimicrobials agents are always up-to-date. The tetrazole derivatives are quite interesting objects as for synthesis as well as for pharmacological screening. Thus, some derivatives of tetrazole demonstrated antimicrobial activity, namely 5-phenyl-tetrazolo[1,5-c]quinazoline was effective one against Staphylococcus aureus and Esherichia faecalis (MIC = 250 mg/L). Besides, investigation of the 9-bromo(chloro)-5-morpholin(piperidine)-4-yl-tetrazolo[1,5-c]quinazoline’s antimicrobial activity against Esherichia coli and Enterococcus faecalis, Pseudomonas aeruginosa and Staphylococcus aureus revealed that sensitivity of Gram-positive bacteria to the compounds was higher than that of Gram-negative bacteria. So, our previously synthesized, 31 derivatives of 1-(2-(1H-tetrazol-5-yl)-R1-phenyl)-3-R2-phenyl(ethyl)ureas were decided to test for their in vitro antibacterial activity against Gram-positive bacteria (Staphylococcus aureus ATCC 25923, Enterobacter aerogenes, Enterococcus faecalis ATCC 29212), Gram-negative bacteria (Pseudomonas aeruginosa ATCC 9027, Escherichia coli ATCC 25922, Klebsiella pneumoniae 68) and antifungal properties against Candida albicans ATCC 885653. Agar-diffusion method was used for determination of the preliminary activity compared to well-known reference antimicrobials. All the compounds were dissolved in DMSO at a concentration of 100 μg/disk, using inhibition zone diameter (IZD, mm) as a measure for the antimicrobial activity. The most active turned to be 3 structures, that inhibited several bacterial strains: 1-ethyl-3-(5-fluoro-2-(1H-tetrazol-5-yl)phenyl)urea (1), 1-(4-bromo-2-(1H-tetrazol-5-yl)-phenyl)-3-(4-(trifluoromethyl)phenyl)urea (2) and 1-(4-chloro-2-(1H-tetrazol-5-yl)phenyl)-3-(3-(trifluoromethyl)phenyl)urea (3). IZM (mm) was 40 (Escherichia coli), 25 (Klebsiella pneumonia) for compound 1; 12 (Pseudomonas aeruginosa), 15 (Staphylococcus aureus), 10 (Enterococcus faecalis) for compound 2; 25 (Staphylococcus aureus), 15 (Enterococcus faecalis) for compound 3. The most sensitive to the activity of the substances were Gram-negative bacteria Pseudomonas aeruginosa. While none of compound effected on Candida albicans. Speaking about, reference drugs: Amikacin (30 µg/disk) showed 27 and Ceftazide (30 µg/disk) 25 against Pseudomonas aeruginosa. That is, unfortunately, higher than studied 1-(2-(1H-tetrazol-5-yl)-R1-phenyl)-3-R2-phenyl(ethyl)ureas. Obtained results will be used for further purposeful optimization of the leading compounds in the more effective antimicrobials because of the ever-mounting problem of microorganism’s resistance.

Keywords: antimicrobial, antifungal, compounds, 1-(2-(1H-tetrazol-5-yl)-R1-phenyl)-3-R2-phenyl(ethyl)ureas

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Authors: Nirupama Sam

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COVID-19, the greatest pandemic since the turn of the century, presents several issues for urban planners, the most significant of which is determining appropriate mitigation techniques for creating pandemic-friendly and resilient public spaces. The study is conducted in four stages. The first stage consisted of literature reviews to examine the evolution and transformation of public spaces during pandemics throughout history and the role of public spaces during pandemic outbreaks. The second stage is to determine the factors that influence the success of public spaces, which was accomplished by an analysis of current literature and case studies. The influencing factors are categorized under comfort and images, uses and activity, access and linkages, and sociability. The third stage is to establish the priority of identified factors for which a questionnaire survey of stakeholders is conducted and analyzing of certain factors with the help of GIS tools. COVID-19 has been in effect in India for the last two years. Kerala has the highest daily COVID-19 prevalence due to its high population density, making it more susceptible to viral outbreaks. Despite all preventive measures taken against COVID-19, Kerala remains the worst-affected state in the country. Finally, two live case studies of the hardest-hit localities, namely Subhash bose park and Napier Museum park in the Ernakulam and Trivandrum districts of Kerala, respectively, were chosen as study areas for the survey. The responses to the questionnaire were analyzed using SPSS for determining the weights of the influencing factors. The spatial success of the selected case studies was examined using the GIS interpolation model. Following the overall assessment, the fourth stage is to develop strategies and guidelines for planning public spaces to make them more efficient and robust, which further leads to improved quality, safety and resilience to future pandemics.

Keywords: urban design, public space, covid-19, post-pandemic, public spaces

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Authors: Akemi Nasu, Keiko Matsumoto

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Rehabilitation for cancer requires setting up individual goals for each patient and an approach that properly fits the stage of cancer when putting into practice. In order to cope with the daily changes in the patients' condition, the establishment of a good cooperative relationship between the nurses and the physiotherapists, occupational therapists, and speech-language-hearing therapists (therapists) becomes essential. This study will focus on the present situation of the cooperation between nurses and therapists, especially the speech-language-hearing therapists, and aim to elucidate what develops there. A semi-structured interview was conducted targeted at a physical therapist having practical experience in working in collaboration with nurses. The contents of the interview were transcribed and converted to data, and the data was encoded and categorized with sequentially increasing degrees of abstraction to conduct a qualitative explorative factor analysis of the data. When providing ethical explanations, particular care was taken to ensure that participants would not be subjected to any disadvantages as a result of participating in the study. In addition, they were also informed that their privacy would be ensured and that they have the right to decline to participate in the study. In addition, they were also informed that the results of the study would be announced publicly at an applicable nursing academic conference. This study has been approved following application to the ethical committee of the university with which the researchers are affiliated. The survey participant is a female speech-language-hearing therapist in her forties. As a result of the analysis, 6 categories were extracted consisting of 'measures to address appetite and aspiration pneumonia prevention', 'limitation of the care a therapist alone could provide', 'the all-inclusive patient- supportive care provided by nurses', 'expand the beneficial cooperation with nurses', 'providing education for nurses on the swallowing function utilizing videofluoroscopic examination of swallowing', 'enhancement of communication including conferences'. In order to improve the team performance, and for the teamwork competency necessary for the provision of safer care, mutual support is essential. As for the cooperation between nurses and therapists, this survey indicates that the maturing of the cooperation between professionals in order to improve nursing professionals' knowledge and enhance communication will lead to an improvement in the quality of the rehabilitation for cancer.

Keywords: cancer rehabilitation, nurses, speech-language-hearing therapists, interprofessional work

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Authors: B. Baba Ahmed, P. Yanogo, B. Djibryl. N. Medas

Abstract:

Introduction: Polio is a highly contagious viral infection, with children under 5 years of age being the most affected. It is a public health emergency of international concern. Polio surveillance in Mauritania has been ongoing since 1998 and has achieved "polio free" status in 2007. our objective is to analyse a pidemiological surveillance database of poliomyélitis in Mauritania from 2012-2019. Method: A transversal descriptive analysis of poliomyélitis database was carried out in Mauritania from 2012-2019.An exhaustive sampling was done on all suspected polio cases recorded in the database from 2012 -2019. This study used EPI-INFO 7.4 for frequency calculation for qualitative variables, mean and standard deviation for quantitative variables. Results: We found 459 suspected cases of polio over the study period with an average rate of acute non-polio flaccid paralysis of 25.4 cases/100,000 children under 15 years of age. The age group 0-6 years represented 75.2%. Males constituted 50.2%. Females represented 49.78% with a ratio of M/F=1.Among the 422 observations, the average age is 4 years +/- 3.38. The four regions, TIRIS-ZEMMOUR, INCHIRI, TAGANT, NOUACHCHOTT OUEST recorded the lowest percentages of notifications, respectively (3.28%; 3.93%; 4.37%; 4.8%). 99.34% [98.09-99.78] of cases presented acute flaccid paralysis. And 56.77% [52.19-61.23], had limb asymmetry. We showed that 82.93% [79.21-86.10], had fever. we found that 89.5% of suspected polio cases were investigated before 48 hours. And 88.39% of suspected cases had two adequate samples taken 48 hours apart and within 14 days after the onset of symptoms. Only 30.95% of samples arrived at the referral laboratory before 72 hours. Conclusion: This study has shown that Mauritania has achieved the objectives in most of the quantitative performance indicators of polio surveillance. This study has shown a low notification of cases in the northern and central regions of the country. There is a problem with the transport of samples to the laboratory.

Keywords: analysis, data base, Epi-Info, polio

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Authors: Musa Sekamatte

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Background: Zoonotic diseases continue to be a public health burden in countries around the world. Uganda is especially vulnerable due to its location, biodiversity, and population. Given these concerns, the Ugandan government in collaboration with the Global Health Security Agenda conducted a zoonotic disease prioritization workshop to identify zoonotic diseases of concern to multiple Ugandan ministries. Materials and Methods: The One Health Zoonotic Disease Prioritization tool, developed by the U.S. Centers for Disease Control and Prevention (CDC), was used for prioritization of zoonotic diseases in Uganda. Workshop participants included voting members representing human, animal, and environmental health ministries as well as key partners who observed the workshop. Over 100 articles describing characteristics of these zoonotic diseases were reviewed for the workshop. During the workshop, criteria for prioritization were selected, and questions and weights relevant to each criterion were determined. Next steps for multi-sectoral engagement for the prioritized zoonoses were then discussed. Results: 48 zoonotic diseases were considered during the workshop. Criteria selected to prioritize zoonotic diseases in order of importance were (1) severity of disease in humans in Uganda, (2) availability of effective control strategies, (3) potential to cause an epidemic or pandemic in humans or animals, (4) social and economic impacts, and (5) bioterrorism potential. Seven zoonotic diseases were identified as priorities for Uganda: anthrax, zoonotic influenza viruses, viral hemorrhagic fevers, brucellosis, African trypanosomiasis, plague, and rabies. Discussion: One Health approaches and multi-sectoral collaborations are crucial in the surveillance, prevention, and control strategies for zoonotic diseases. Uganda used such an approach to identify zoonotic diseases of national concern. Identifying these priority diseases enables the National One Health Platform and the Zoonotic Disease Coordinating Office to address the diseases in the future.

Keywords: national one health platform, zoonotic diseases, multi-sectoral, severity

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Authors: Mohammad K. Parvez, Mohammed S. Al-Dosari

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Hepatitis E virus (HEV) is an emerging RNA virus that causes acute and chronic liver disease with a global mortality rate of about 2%. Despite milestone developments in understanding of HEV biology, there is still lack of a robust culture system or animal model. Therefore, in a novel approach, two recombinant-baculoviruses (vBac-ORF2 and vBac-ORF3) that could overexpress HEV ORF2 (structural/capsid) and ORF3 (nonstructural/regulatory) proteins, respectively were constructed. The established HEV-SAR55 (genotype 1) replicon that contained GFP gene, in place of ORF2/ORF3 sequences was in vitro transcribed, and GFP production in RNA transfected S10-3 cells was scored by FACS. Enhanced infectivity, if any, of nascent virions produced by exogenously-supplied ORF2 and viral RNA by co-expression of ORF3 was tested on naïve HepG2 cells. Co-transduction with vBac-ORF2/vBac-ORF3 (108 pfu/microL) produced high amounts of native ORF2/ORF3 in approximately 60% of S10-3 cells, determined by immunofluorescence microscopy and Western analysis. FACS analysis showed about 9% GFP positivity of S10-3 cells on day6 post-transfection (i.e, day5 post-transduction). Further, FACS scoring indicated that lysates from S10-3 cultures receiving the RNA plus vBac-ORF2 were capable of producing HEV particles with about 4% infectivity in HepG2 cells. However, lysates of cultures co-transduced with vBac-ORF3, were found to further enhance virion infectivity by approximately 17%. This supported a previously proposed role of ORF3 as a minor-structural protein in HEV virion assembly and infectivity. In conclusion, the present model for efficient genomic RNA packaging and production of infectious virions could be a valuable tool to study various aspects of HEV molecular biology, in vitro.

Keywords: chronic liver disease, hepatitis E virus, ORF2, ORF3, replicon

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Authors: Inam Ridha, Christine L. Kuryla, Madhuranga Thilakasiri Madugoda Ralalage Don, Norman J. Kleiman, Yunro Chung, Jin Park, Vel Murugan, Joshua LaBaer

Abstract:

To date, more than 275 million people worldwide have been diagnosed with COVID-19 and the rapid spread of the omicron variant suggests many millions more will soon become infected. Many infections are asymptomatic, while others result in mild to moderate illness. Unfortunately, some infected individuals exhibit more serious symptoms including respiratory distress, thrombosis, cardiovascular disease, multi-organ failure, cognitive difficulties, and, in roughly 2% of cases, death. Studies indicate other coronaviruses can alter the host cell's epigenetic profile and lead to alterations in the immune response. To better understand the mechanism(s) by which SARS-CoV-2 infection causes serious illness, DNA methylation profiles in peripheral blood mononuclear cells (PBMCs) from 90 hospitalized severely ill COVID-19 patients were compared to profiles from uninfected control subjects. Exploratory epigenome-wide DNA methylation analyses were performed using multiplexed methylated DNA immunoprecipitation (MeDIP) followed by pathway enrichment analysis. The findings demonstrated significant DNA methylation changes in infected individuals as compared to uninfected controls. Pathway analysis indicated that apoptosis, cell cycle control, Toll-like receptors (TLR), cytokine interactions, and T cell differentiation were among the most affected metabolic processes. In addition, changes in specific gene methylation were compared to SARS-CoV-2 induced changes in RNA expression using published RNA-seq data from 3 patients with severe COVID-19. These findings demonstrate significant correlations between differentially methylated and differentially expressed genes in a number of critical pathways.

Keywords: COVID19, epigenetics, DNA mathylation, viral infection

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Authors: Yu-Chieh Chen, Li-Yun Wang, Chi-Chih Chen, Huy Hùng Đào, Ya-Mei Chen, Ming-Chu Cheng, Wen-Bin Chung, Hso-Chi Chaung, Guan-Ming Ke

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Many recent researches have demonstrated that CD154, a protein primarily expressed on activated T cell molecules, has potentially acted as a molecular adjuvant to improve the immunogenicity of subunit vaccines against viral infections. Classical swine fever (CSF) affects the swine industry worldwide that is one of the most devastating and highly contagious pig diseases. It is listed by the World Organization for Animal Health (OIE) as an infectious animal disease that must be reported. Although pigs vaccinated with subunit vaccines can be differentially diagnosed from those infected animals, subunit vaccines usually need adjuvants to enhance and elicit immune responses. In this study, CD154 was linked with CSFV E2 sequences and then expressed in CHO cells to produce the fusion protein as E2-CD154. The porcine specific CpG adjuvant was also used in one of the formulations. The specific pathogen-free pigs (SPF) at the age of 4-week-old were randomly separated into four groups, vaccinated with E2-CpG, E2-CD154, E2-CD154-CpG or the commercial Bayovac® CSF-E2 vaccine and boosted two weeks after primary vaccination. The results showed that the percentages of CD4+ and CD4+IL2+ in peripheral blood mononuclear cells (PBMC) in E2-CD154 vaccinated piglets seven days after primary vaccination were gained by 1-5% relative to the control group. In addition, the percentages of CD4+IFNγ+ T cells had slightly edged up 0.1-0.3% compared with the control group. Also, increased E2-specific IFNγ levels had edged up CD4+CD8+ T cells found in E2-CD154 and E2-CD154-CpG groups, particularly in the E2-CD154-CpG group. These results implicate that CD154 may enhance cellular immunity and synergistically act with species-specific CpG adjuvant as a dual-phase adjuvant. Therefore, the CD154 may be beneficial as a promising adjuvant in subunit vaccines.

Keywords: CD154, CpG adjuvant, cellular immunity, subunit vaccine, pig

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Authors: Hakan Duger, Alparslan Ersoy, Canan Ersoy

Abstract:

Diabetes mellitus is the most common etiology of end-stage renal disease (ESRD). Also, diabetic nephropathy is the etiology of ESRD in approximately 23% of kidney transplant recipients. A successful kidney transplant improves the quality of life and reduces the mortality risk for most patients. However, patients require close follow-up after transplantation due to medical complications. Diabetes mellitus can affect patient morbidity and mortality due to possible effects of immunosuppressive therapy on glucose metabolism. We compared the frequency of medical complications and the outcomes in diabetic and non-diabetic kidney transplant recipients. Materials and Methods: This retrospective study conducted in 498 patients who underwent kidney transplant surgery at our center in 10-year periods. The patients were divided into two groups: diabetics (46 ± 10 year, 26 males, 16 females) and non-diabetics (39 ± 12 year, 259 males, 197 females). The medical complications, graft functions, causes of graft loss and death were obtained from medical records. Results: There was no significant difference between recipient age, duration of dialysis, body mass index, gender, donor type, donor age, dialysis type, histories of HBV, HCV and coronary artery disease between two groups. The history of hypertension in diabetics was higher (69% vs. 36%, p < 0.001). The ratios of hypertension (50.1% vs. 57.1%), pneumonia (21.9% vs. 20%), urinary infection (16.9% vs. 20%), transaminase elevation (11.5% vs. 20%), hyperpotasemia (14.7% vs. 17.1%), hyponatremia (9.7% vs. 20%), hypotension (7.1% vs. 7.9%), hypocalcemia (1.4% vs. 0%), thrombocytopenia (8.6% vs. 8.6%), hypoglycemia (0.7% vs. 0%) and neutropenia (1.8% vs. 0%) were comparable in non-diabetic and diabetic groups, respectively. The frequency of hyperglycaemia in diabetics was higher (8.6% vs. 54.3%, p < 0.001). After transplantation, primary non-function (3.4% vs. 2.6%), delayed graft function (25.1% vs. 34.2%) and acute rejection (7.3% vs. 10.5%) ratios of in non-diabetic and diabetic groups were similar, respectively. Hospitalization durations in non-diabetics and diabetics were 22.5 ± 17.5 and 18.7 ± 13 day (p=0.094). Mean serum creatinine levels in non-diabetics and diabetics were 1.54 ± 0.74 and 1.52 ± 0.62 mg/dL at 6th month. Forty patients had graft loss. The ratios of graft loss and death in non-diabetic and diabetic groups were 8.2% vs. 7.1% and 7.1% vs. 2.6% (p > 0.05). There was no significant relationship between graft and patient survivals with the development of medical complication. Conclusion: As a result, medical complications are common in the early period. Hyperglycaemia was frequently seen following transplantation due to the effects of immunosuppressant regimens. However, the frequency of other medical complications in diabetic patients did not differ from non-diabetic one. The most important cause of death is still infections. The development of medical complications during the first 6 months did not significantly affect transplant outcomes.

Keywords: kidney transplantation, diabetes mellitus, complication, graft function

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Authors: Dharmendra Pratap

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Large Cardamom (Amomum subulatum), family Zingiberaceae is an aromatic spice crop and has rich medicinal value. Large Cardamom is as synonymous to Sikkim as Tea is to Darjeeling. Since Sikkim alone contributes up to 88% of India's large cardamom production which is the world leader by producing over 50% of the global yield. However, the production of large cardamom has declined almost to half since last two decade. The economic losses have been attributed to two viral diseases namely, chirke and Foorkey. Chirke disease is characterized by light and dark green streaks on leaves. The affected leaves exhibit streak mosaic, which gradually coalesce, turn brown and eventually dry up. Excessive sprouting and formation of bushy dwarf clumps at the base of mother plants that gradually die characterize the foorkey disease. In our surveys in Sikkim–Darjeeling hill area during 2012-14, 40-45% of plants were found to be affected with foorkey disease and 10-15% with chirke. Mechanical and aphid transmission study showed banana as an alternate host for both the disease. For molecular identification, total genomic DNA and RNA was isolated from the infected leaf tissues and subjected to Rolling circle amplification (RCA) and RT-PCR respectively. The DNA concatamers produced in the RCA reaction were monomerized by different restriction enzymes and the bands corresponding to ~1 kb genomes were purified and cloned in the respective sites. The nucleotide sequencing results revealed the association of Nanovirus with the foorkey disease of large cardamom. DNA1 showed 74% identity with Replicase gene of FBNYV, DNA2 showed 77% identity with the NSP gene of BBTV and DNA3 showed 74% identity with CP gene of BBTV. The finding suggests the presence of a new species of nanovirus associated with foorkey disease of large cardamom in Sikkim and Darjeeling hills. The details of their epidemiology and other factors would be discussed.

Keywords: RCA, nanovirus, large cardamom, molecular virology and microbiology

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Authors: Abhaydeep Pandey, Shweta Shukla, Saptamita Goswami, Bhaswati Bandyopadhyay, Vishnampettai Ramachandran, Sudhanshu Vrati, Arup Banerjee

Abstract:

Background: Long non-coding RNAs (lncRNAs) are the important regulators of gene expression and play important role in viral replication and disease progression. The role of lncRNA genes in the pathogenesis of Dengue virus-mediated pathogenesis is currently unknown. Methods: To gain additional insights, we utilized an unbiased RNA sequencing followed by in silico analysis approach to identify the differentially expressed lncRNA and genes that are associated with dengue disease progression. Further, we focused our study on lncRNAs NEAT1 (Nuclear Paraspeckle Assembly Transcript 1) as it was found to be differentially expressed in PBMC of dengue infected patients. Results: The expression of lncRNAs NEAT1, as compared to dengue infection (DI), was significantly down-regulated as the patients developed the complication. Moreover, pairwise analysis on follow up patients confirmed that suppression of NEAT1 expression was associated with rapid fall in platelet count in dengue infected patients. Severe dengue patients (DS) (n=18; platelet count < 20K) when recovered from infection showing high NEAT1 expression as it observed in healthy donors. By co-expression network analysis and subsequent validation, we revealed that coding gene; IFI27 expression was significantly up-regulated in severe dengue cases and negatively correlated with NEAT1 expression. To discriminate DI from dengue severe, receiver operating characteristic (ROC) curve was calculated. It revealed sensitivity and specificity of 100% (95%CI: 85.69 – 97.22) and area under the curve (AUC) = 0.97 for NEAT1. Conclusions: Altogether, our first observations demonstrate that monitoring NEAT1and IFI27 expression in dengue patients could be useful in understanding dengue virus-induced disease progression and may be involved in pathophysiological processes.

Keywords: dengue, lncRNA, NEAT1, transcriptome

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