Search results for: infectious salmon anemia virus

Authors: Najmeh Jafari, Sona Rostampour Yasouri

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Crimean-Congo hemorrhagic fever (CCHF) is an acute hemorrhagic disease. This disease is one of the common diseases between humans and animals, transmitted through tick bites or contact with the blood and secretions or carcasses of infected animals and humans. CCHF is more common in people who work with livestock, such as ranchers, butchers, farmers, slaughterhouse workers, healthcare workers, etc. Its hospital prevalence is also very high. Considering that CCHF can be transmitted through the consumption of food such as beef and sheep meat, this study aims to quickly identify and diagnose the Crimean-Congo fever virus in suspected patients through real-time PCR technique. In the summer of 1402, 20 blood samples were collected separately from Ahvaz and Gilan hospitals. An extraction kit was used to extract the virus RNA. Primers and probes were designed based on the S genomic region, the conserved region in CCHFV. Then, a real-time PCR technique was performed with specific primers and probes. It should be noted that the mentioned technique was repeated several times. The number of 4 samples from the examined samples was determined positive by real-time PCR. This technique has high sensitivity and specificity and the possibility of rapid detection of CCHFV. Therefore, the above method is a good candidate for quick disease diagnosis. By diagnosing the disease, the treatment process can be done faster, and the best prevention methods can be used to control the disease and prevent the death of patients.

Keywords: ahvaz, crimean-congo hemorrhagic fever, gilan, real time PCR

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Authors: Laura Amato, Paolo Mulatti, Fabrizio Montarsi, Matteo Mazzucato, Laura Gagliazzo, Michele Brichese, Manlio Palei, Gioia Capelli, Lebana Bonfanti

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West Nile virus (WNV) re-emerged in north-eastern Italy in 2008, after ten years from its first appearance in Tuscany. In 2009, a national surveillance programme was implemented, and re-modulated in north-eastern Italy in 2011. Hereby, we present the results of surveillance activities in 2008-2016 in the north-eastern Italian regions, with inferences on WNV epidemiological trend in the area. The re-modulated surveillance programmes aimed at early detecting WNV seasonal reactivation by searching IgM antibodies in horses. In 2013, the surveillance plans were further modified including a risk-based approach. Spatial analysis techniques, including Bernoulli space-time scan-statistics, were applied to the results of 2010–2012 surveillance on mosquitoes, equines, and humans to identify areas where WNV reactivation was more likely to occur. From 2008 to 2016, residential horses tested positive for anti-WNV antibodies on a yearly basis (503 cases), also in areas where WNV circulation was not detected in mosquito populations. Surveillance activities detected 26 syndromic cases in horses, 102 infected mosquito pools and WNV in 18 dead wild birds. Human cases were also recurrently detected in the study area during the surveillance period (68 cases of West Nile neuroinvasive disease). The recurrent identification of WNV in animals, mosquitoes, and humans indicates the virus has likely become endemic in the area. In 2016, findings of WNV positives in horses or mosquitoes were included as triggers for enhancing screening activities in humans. The evolution of the epidemiological situation prompts for continuous and accurate surveillance measures. The results of the 2013-2016 surveillance indicate that the risk-based approach was effective in early detecting seasonal reactivation of WNV, key factor of the integrated surveillance strategy in endemic areas.

Keywords: arboviruses, horses, Italy, surveillance, west nile virus, zoonoses

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Authors: Mehmet Kale, Ramazan Sencan, Sibel Yavru, Ahmet Ak, Nuri Mamak, Sibel Hasırcıoglu, Mesih Kocamuftuoglu, Yakup Yıldırım, Hasbi Sait Saltık

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Bovine Papilloma Virus (BPV)-induced warts can cause mastitis, teat blindness, reduction of milk yield, udder deformities, and a difficulty in getting the teats into the milking machine. Especially, surgical operations cannot be performed in BPV-induced teat warts because of the increased sensitivity of the breast region and small-sized papillomas. Thus, there is a need to find new topical treatment methods. We have developed a pomade for treatment of BPV in cattle. The pomade is consists of lanoline, snakeskin (two special kind of snake), alcohol, vaseline, and ether. Firstly, we determined 46 cattle with teat warts. In the study, BPV antigen was detected in 28 cattle blood samples (61%) by ELISA. The pomade was applied to all BPV infected animals. The regression and recovery of warts were 100% in all animals. We advised using the pomade for treatment of BPV-induced warts in teats.

Keywords: bovine papilloma virus, pomade, teat, udder

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Authors: Soma Banerjee, Aamen Talukdar, Argha Mandal, Dipankar Chaudhuri

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Japanese Encephalitis is a major infectious disease with nearly half the world’s population living in areas where it is prevalent. Currently, treatment for it involves only supportive care and symptom management through vaccination. Due to the lack of antiviral drugs against Japanese Encephalitis Virus (JEV), the quest for such agents remains a priority. For these reasons, simulation studies of drug targets against JEV are important. Towards this purpose, docking experiments of the kinase inhibitors were done against the chosen target NS3 helicase as it is a nucleoside binding protein. Previous efforts regarding computational drug design against JEV revealed some lead molecules by virtual screening using public domain software. To be more specific and accurate regarding finding leads, in this study a proprietary software Schrödinger-GLIDE has been used. Druggability of the pockets in the NS3 helicase crystal structure was first calculated by SITEMAP. Then the sites were screened according to compatibility with ATP. The site which is most compatible with ATP was selected as target. Virtual screening was performed by acquiring ligands from databases: KinaseSARfari, KinaseKnowledgebase and Published inhibitor Set using GLIDE. The 25 ligands with best docking scores from each database were re-docked in XP mode. Protein structure alignment of NS3 was performed using VAST against MMDB, and similar human proteins were docked to all the best scoring ligands. The low scoring ligands were chosen for further studies and the high scoring ligands were screened. Seventy-three ligands were listed as the best scoring ones after performing HTVS. Protein structure alignment of NS3 revealed 3 human proteins with RMSD values lesser than 2Å. Docking results with these three proteins revealed the inhibitors that can interfere and inhibit human proteins. Those inhibitors were screened. Among the ones left, those with docking scores worse than a threshold value were also removed to get the final hits. Analysis of the docked complexes through 2D interaction diagrams revealed the amino acid residues that are essential for ligand binding within the active site. Interaction analysis will help to find a strongly interacting scaffold among the hits. This experiment yielded 21 hits with the best docking scores which could be investigated further for their drug like properties. Aside from getting suitable leads, specific NS3 helicase-inhibitor interactions were identified. Selection of Target modification strategies complementing docking methodologies which can result in choosing better lead compounds are in progress. Those enhanced leads can lead to better in vitro testing.

Keywords: antivirals, docking, glide, high-throughput virtual screening, Japanese encephalitis, ns3 helicase

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Authors: Muhammad Asif Syed, Mirza Amir Baig

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Background: Pakistan is one of the two remaining polio-endemic countries, posing a significant public health challenge for global polio eradication due to failure to interrupt polio transmission. Country specific seroprevalence studies help in the evaluation of immunization program performance, the susceptibility of population against polio virus and identification of existing level of immunity with factors that affect seroconversion of the oral polio vaccine (OPV). The objective of the study was to find out factors associated with seroconversion of the OPV among children 6-59 months in Pakistan. Methods: A Hospital based cross-sectional serosurvey was undertaken in May-June 2015 at District Mirpurkhas, Sindh-Pakistan. Total 180 children aged 6–59 months were selected by using systematic random sampling from Muhammad Medical College Hospital, Mirpurkhas. Demographic, vaccination history and risk factors information were collected from the parents/guardian. Blood sample was collected and tested for the detection of poliovirus IgG antibodies by using ELISA Kit. The IgG titer <10 IU/ml, 50 to <150 IU/ml and >150 IU/ml was defined as negative, weak positive and positive immunity respectively. Pearson Chi-square test was used to determine the difference in seroprevalence in univariate analysis. Results: A total of 180 subjects were enrolled mean age was 23 months (7 -59 months). Off these 160 (89%) children were well and 18 (10%) partially protected against polio virus. Two (1.1%) children had no protection against polio virus as they had <10 IU/ml poliovirus IgG antibodies titer. Both negative cases belong from the female gender, age group 12-23 months, urban area and BMI <50 percentile. There was a difference between normal and the wasting children; it did attain statistical significance (χ2= 35.5, p=0.00). The difference in seroconversion was also observed in relation to the gender (χ2=6.23, p=0.04), duration of breast feeding (χ2=18.6, p=0.04), history of diarrheal disease before polio vaccine administration (χ2=7.7, p=0.02), and stunting (χ2= 114, p=0.00). Conclusion: This study demonstrated that near 90% children achieve seroconversion of OPV and well protected against polio virus. There is an urgent need to focus on factors like duration of breast feeding, diarrheal diseases and malnutrition (acute and chronic) among the children as an immunization strategy.

Keywords: seroconversion, oral polio vaccine, Polio, Pakistan

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Authors: Rakesh Kumar Meena, Tanushree Chatterjee

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Okra (Abelmoschus esculentas L. Moench) or lady’s finger is an important vegetable crop belonging to the Malvaceae family. Unfortunately, production and productivity of Okra are majorly affected by Yellow Vein mosaic virus (YVMV). The AO: 189 (resistant parent) X AO: 191(susceptible parent) used for the development of mapping population. The mapping population has 143 individuals (F₂:F₃). Population was characterized by physiological and pathological observations. Screening of 360 DNA markers was performed to survey for parental polymorphism between the contrasting parents’, i.e., AO: 189 and AO: 191. Out of 360; 84 polymorphic markers were used for genotyping of the mapping population. Total markers were distributed into four linkage groups (LG1, LG2, LG3, and LG4). LG3 covered the longest span (106.8cM) with maximum number of markers (27) while LG1 represented the smallest linkage group in terms of length (71.2cM). QTL identification using the composite interval mapping approach detected two prominent QTLs, QTL1 and QTL2 for resistance against YVMV disease. These QTLs were placed between the marker intervals of NBS-LRR72-Path02 and NBS-LRR06- NBS-LRR65 on linkage group 02 and linkage group 04 respectively. The LOD values of QTL1 and QTL2 were 5.7 and 6.8 which accounted for 19% and 27% of the total phenotypic variation, respectively. The findings of this study provide two linked markers which can be used as efficient diagnostic tools to distinguish between YVMV resistant and susceptible Okra cultivars/genotypes. Lines identified as highly resistant against YVMV infection can be used as donor lines for this trait. This will be instrumental in accelerating the trait improvement program in Okra and will substantially reduce the yield losses due to this viral disease.

Keywords: Okra, yellow vein mosaic virus, resistant, linkage map, QTLs

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Authors: H. Boulahyaoui, S. Alaoui Amine, C. Loutfi, H. El Annaz, N. Touil, El M. El Fahim, S. Mrani

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Three Moroccan children fully vaccinated with Rotarix™ have been hospitalized for Rotavirus Gastroenteritis (RVGE) in the pediatric division of the Farabi Hospital, Oujda. Rotavirus G/P genotypes could not be typed because of their delayed crossing threshold (Ct) resolute with a group A rotavirus (RVA) real time RT-PCR. These strains were adapted to cell culture. All viruses replicated and caused extensive cytopathic effects after four or five passages in MA104 cell lines. Significant improvements have been obtained in the amount of viral particles. Each virus multiplied to a high titer (7.5 TCID50/ml). VP7 and VP4 partial gene sequencing revealed distinct genotypes compared to the Rotarix(®) vaccine strain. Two strains were of G2P[4] genotype whereas the third was G9P[8] genotype. Virus isolation while labor intensive, is recommended as a second test, especially when higher sensitivity for conventional RVA genotyping RT-PCR is needed. VP7 antigenic similarities between these strains and Rotarix were determined.

Keywords: esacpe-vaccine, Morocco, Rotarix, G2P[4], G9P[8]

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Authors: Atlal El-Assaad

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Viruses change with time through mutations and result in new variants that may persist or disappear. A Mutation refers to an actual change in the virus genetic sequence, and a variant is a viral genome that may contain one or more mutations. Critical mutations may cause the virus to be more transmissible, with high disease severity, and more vulnerable to diagnostics, therapeutics, and vaccines. Thus, variants carrying such mutations may increase the risk to human health and are considered variants of concern (VOC). Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) - the contagious in humans, positive-sense single-stranded RNA virus that caused coronavirus disease 2019 (COVID-19) - has been studied thoroughly, and several variants were revealed across the world with their corresponding mutations. SARS-CoV-2 has four structural proteins, known as the S (spike), E (envelope), M (membrane), and N (nucleocapsid) proteins, but prior study and vaccines development focused on genetic mutations in the S protein due to its vital role in allowing the virus to attach and fuse with the membrane of a host cell. Specifically, subunit S1 catalyzes attachment, whereas subunit S2 mediates fusion. In this perspective, we studied all charged amino acid mutations of the SARS-CoV-2 viral spike protein S1 when bound to Antibody CC12.1 in a crystal structure and assessed the effect of different mutations. We generated all missense mutants of SARS-CoV-2 protein amino acids (AAs) within the SARS-CoV-2:CC12.1 complex model. To generate the family of mutants in each complex, we mutated every charged amino acid with all other charged amino acids (Lysine (K), Arginine (R), Glutamic Acid (E), and Aspartic Acid (D)) and studied the new binding of the complex after each mutation. We applied Poisson-Boltzmann electrostatic calculations feeding into free energy calculations to determine the effect of each mutation on binding. After analyzing our data, we identified charged amino acids keys for binding. Furthermore, we validated those findings against published experimental genetic data. Our results are the first to propose in silico potential life-threatening mutations of SARS-CoV-2 beyond the present mutations found in the five common variants found worldwide.

Keywords: SARS-CoV-2, variant, ionic amino acid, protein-protein interactions, missense mutation, AESOP

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Authors: José Silvestre Mendoza Figueroa, Anders Kvarnheden, Jesús Méndez Lozano, Edgar Antonio Rodríguez Negrete, Manuel Soriano García

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Agroindustrial plants such as cereals and pseudo cereals offer a substantial source of biomacromolecules, as they contain large amounts per tissue-gram of proteins, polysaccharides and lipids in comparison with other plants. In particular, Amaranthus hypochondriacus seeds have high levels of proteins in comparison with other cereal and pseudo cereal species, which makes the plant a good source of bioactive molecules such as peptides. Geminiviruses are one principal class of pathogens that causes important economic losses in crops, affecting directly the development and production of the plant. One such virus is the Tomato yellow leaf curl virus (TYLCV), which affects mainly Solanacea family plants such as tomato species. The symptoms of the disease are curling of leaves, chlorosis, dwarfing and floral abortion. The aim of this work was to get peptides derived from enzymatic hydrolysis of globulins and albumins from amaranth seeds with specific recognition of the replication origin in the TYLCV genome, and to test the antiviral activity on host plants with the idea to generate a direct control of this viral infection. Globulins and albumins from amaranth were extracted, the fraction was enzymatically digested with papain, and the aromatic peptides fraction was selected for further purification. Six peptides were tested against the replication origin (OR) using affinity assays, surface resonance plasmon and fluorescent titration, and two of these peptides showed high affinity values to the replication origin of the virus, dissociation constant values were calculated and showed specific interaction between the peptide Ampep1 and the OR. An in vitro replication test of the total TYLCV DNA was performed, in which the peptide AmPep1 was added in different concentrations to the system reaction, which resulted in a decrease of viral DNA synthesis when the peptide concentration increased. Also, we showed that the peptide can decrease the complementary DNA chain of the virus in Nicotiana benthamiana leaves, confirming that the peptide binds to the OR and that its expected mechanism of action is to decrease the replication rate of the viral genome. In an infection assay, N. benthamiana plants were agroinfected with TYLCV-Israel and TYLCV-Guasave. After confirming systemic infection, the peptide was infiltrated in new infected leaves, and the plants treated with the peptide showed a decrease of virus symptoms and viral titer. In order to confirm the antiviral activity in a commercial crop, tomato plants were infected with TYLCV. After confirming systemic infection, plants were infiltrated with peptide solution as above, and the symptom development was monitored 21 days after treatment, showing that tomato plants treated with peptides had lower symptom rates and viral titer. The peptide was also tested against other begomovirus such as Pepper huasteco yellow vein virus (PHYVV-Guasave), showing a decrease of symptoms in N. benthamiana infected plants. The model of direct biochemical control of TYLCV infection shown in this work can be extrapolated to other begomovirus infections, and the methods reported here can be used for design of antiviral agrochemicals for other plant virus infections.

Keywords: agrochemical screening, antiviral, begomovirus, geminivirus, peptides, plasmon, TYLCV

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Authors: I. L. Ibrahim, A. Mann, B. M. Abdullahi

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Infectious diseases are prevalent in developing countries and plant extracts are known to contained bioactive compounds that can be used in the management of these diseases. The entire plant of Chrysanthellum indicum (Linn) was air-dried and pulverized into fine powder and then percolated to give ethanol and aqueous extracts. These extracts were phytochemically screened for metabolites and evaluated antibacterial activity against some pathogenic organisms Klebsilla, pneumonia, Bacillus subtilis, and Pseudomonas aeruginosa using agar dilution method. It was found that crude extracts of C. indicum revealed the presence of saponins, tannins, alkaloids, steroidal nucleus, cardiac glycosides, and coumarin while flavonoids and anthraquinones were absent. The Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of the active extract of C. indicum shows that the extract could be a potential source of antibacterial agents.

Keywords: antibacterial activity, Chrysanthellum indicum, infectious diseases, phytochemical screening

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Authors: Charu Kriti

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The year 2020 will be remembered in history as the year when everything changed suddenly. As the world is engrossed in fighting a pandemic, individual life has taken a hit. The sudden imposition of lockdown, the perpetual fear of testing positive for the COVID virus, and rescheduling one’s entire life around this one global phenomenon have created unprecedented stress among all cadres. This paper aims to highlight the level of stress that students face during the shift of the classroom from the physical setup to the virtual one. The paper takes into account the day-to-day hassles that a student faces during online classes. The paper also attempts to analyse these from the other perspective of the students’ lives and the difficulties faced by them on all fronts. This is an empirical research paper that takes into account responses from a total of 4,241 students. The responses have been collected via the online survey, which is being assessed and inferred for the purposes of this paper. The final results show the extent of stress that online classes have induced upon the students.

Keywords: anxiety, COVID, stress, anxiety disorder

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Authors: Yik Sin Chan, Nam Weng Sit, Fook Yee Chye, van Ofwegen Leen, de Voogd Nicole, Kong Soo Khoo

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Chikungunya fever is an arboviral disease transmitted by the Aedes mosquitoes. It has resulted in epidemics of the disease in tropical countries in the Indian Ocean and South East Asian regions. The recent spread of this disease to the temperate countries such as France and Italy, coupled with the absence of vaccines and effective antiviral drugs make chikungunya fever a worldwide health threat. This study aims to investigate the anti-chikungunya virus activity of selected marine organism samples collected from Malaysian coastal areas, including seaweeds (Caulerpa racemosa, Caulerpa sertularioides and Kappaphycus alvarezii), a soft coral (Lobophytum microlobulatum) and a sponge (Spheciospongia vagabunda). Following lyophilization (oven drying at 40C for K. alvarezii) and grinding to powder form, each sample was subjected to sequential solvent extraction using hexane, chloroform, ethyl acetate, ethanol, methanol and distilled water in order to extract bioactive compounds. The antiviral activity was evaluated using monkey kidney epithelial (Vero) cells infected with the virus (multiplicity of infection=1). The cell viability was determined by Neutral Red uptake assay. 70% of the 30 extracts showed weak inhibitory activity with cell viability ≤30%. Seven of the extracts exhibited moderate inhibitory activity (cell viability: 31%-69%). These were the chloroform, ethyl acetate, ethanol and methanol extracts of C. racemosa; chloroform and ethyl acetate extracts of L. microlobulatum; and the chloroform extract of C. sertularioides. Only the hexane and ethanol extracts of L. microlobulatum showed strong inhibitory activity against the virus, resulting in cell viabilities (mean±SD; n=3) of 73.3±2.6% and 79.2±0.9%, respectively. The corresponding mean 50% effective concentrations (EC50) for the extracts were 14.2±0.2 and 115.3±1.2 µg/mL, respectively. The ethanol extract of the soft coral L. microlobulatum appears to hold the most promise for further characterization of active principles as it possessed greater selectivity index (SI>5.6) compared to the hexane extract (SI=2.1).

Keywords: antiviral, seaweed, sponge, soft coral, vero cell

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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: Ansah Iris Baffour, Dong-Ho Kim, Sung-Gyu Park

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The ongoing COVID-19 pandemic, caused by the SARS-CoV-2 virus and is gradually shifting to an endemic phase which implies the outbreak is far from over and will be difficult to eradicate. Global cooperation has led to unified precautions that aim to suppress epidemiological spread (e.g., through travel restrictions) and reach herd immunity (through vaccinations); however, the primary strategy to restrain the spread of the virus in mass populations relies on screening protocols that enable rapid on-site diagnosis of infections. Herein, we employed surface enhanced Raman spectroscopy (SERS) for the rapid detection of SARS-CoV-2 lysate on an Au-modified Au nanodimple(AuND)electrode. Through in situone-pot Au electrodeposition on the AuND electrode, Au-lysate nanocomposites were synthesized, generating3D internal hotspots for large SERS signal enhancements within 30 s of the deposition. The capture of lysate into newly generated plasmonic nanogaps within the nanocomposite structures enhanced metal-spike protein contact in 3D spaces and served as hotspots for sensitive detection. The limit of detection of SARS-CoV-2 lysate was 5 x 10-2 PFU/mL. Interestingly, ultrasensitive detection of the lysates of influenza A/H1N1 and respiratory syncytial virus (RSV) was possible, but the method showed ultimate selectivity for SARS-CoV-2 in lysate solution mixtures. We investigated the practical application of the approach for rapid on-site diagnosis by detecting SARS-CoV-2 lysate spiked in normal human saliva at ultralow concentrations. The results presented demonstrate the reliability and sensitivity of the assay for rapid diagnosis of COVID-19.

Keywords: label-free detection, nanocomposites, SARS-CoV-2, surface-enhanced raman spectroscopy

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Authors: Sujith Rajashekar Swamy, Meghana Rajashekara Swamy

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Although T1-weighted gadolinium-enhanced imaging (T1-Gd) has its established clinical role in diagnosing brain lesions of infectious and metastatic origins, the use of post-contrast susceptibility-weighted imaging (SWI) has been understudied. This observational study aims to explore and compare the prominence of brain parenchymal lesions between T1-Gd and SWI-Gd images. A cross-sectional study design was utilized to analyze 58 patients with brain parenchymal lesions using T1-Gd and SWI-Gd scanning techniques. Our results indicated that SWI-Gd enhanced the conspicuity of metastatic as well as infectious brain lesions when compared to T1-Gd. Consequently, it can be used as an adjunct to T1-Gd for post-contrast imaging, thereby avoiding additional contrast administration. Improved conspicuity of brain lesions translates directly to enhanced patient outcomes, and hence SWI-Gd imaging proves useful to meet that endpoint.

Keywords: susceptibility weighted, T1 weighted, brain lesions, gadolinium contrast

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Authors: Borys Stegniy, Anton Gerilovych, Oleksii Solodiankin, Vitaliy Bolotin, Anton Stegniy, Denys Muzyka, Claudio Afonso

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New APMV was isolated from white fronted goose in Ukraine. This isolate was tested serologically using monoclonal antibodies in haemagglutination-inhibition tests against APMV1-9. As the results obtained isolate showed cross reactions with APMV7. Following investigations were provided for the full genome sequencing using random primers and cloning into pCRII-TOPO. Analysis of 100 transformed colonies of E.coli using traditional sequencing gave us possibilities to find only 3 regions, which could identify by BLAST. The first region with the length of 367 bp had 70 % nucleotide sequence identity to the APMV 12 isolate Wigeon/Italy/3920_1/2005 at genome position 2419-2784. Next region (344 bp) had 66 % identity to the same APMV 12 isolate at position 4760-5103. The last region (365 bp) showed 71 % identity to Newcastle disease virus strain M4 at position 12569-12928.

Keywords: APMV, Newcastle disease virus, Ukraine, full genome sequencing

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Authors: Mikailzade Parvin, Gurbanova Jamila, Alizade Samaya, Hasanova Afat

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It is known that in March 2020, the World Health Organization (WHO) declared a global pandemic of the 2019 coronovirus disease COVID-19, caused by the severe acute respiratory syndrome coronovirus (SARS-CoV-2). In this period, ensuring the safety of pregnancy and childbirth has become one of the necessary issues. The measures taken in this direction naturally consisted of strengthening and improving preventive measures among pregnant women. It should be noted that the lethality of SARS-CoV-2 infection among women reached 25%. The relevance of studying the effect of COVID-19 on reproductive health in women is due to its wide spread worldwide, severe clinical course, and the occurrence of numerous complications or lethality. It is of urgent importance to study the impact of the mentioned coronavirus infection on the health of pregnant women and the serious complications caused by it.Taking these into account, 230 pregnant women infected with the COVID-19 virus infection were registered. The average age of the pregnant women included in the study was: 29.24±6.0. The diagnosis of corona virus infection was made on the basis of polymerase chain reaction (PCR), serological tests (IgG, IgM). In 57.4% of cases, bilateral pneumonia was recorded in pregnant women and confirmed on the basis of radiological (RH) examination. RH examination revealed pneumonia with infiltrate in the lungs. Among clinical symptoms in pregnant women infected with COVID-19 virus infection: in 86 (37.4%) cases, symptoms such as high fever (t-39.0oC), shortness of breath, fatigue, and hypoxia were noted in pregnant women. A decrease in SpO2 to a minimal level was recorded. Laboratory-instrumental examinations were carried out. The obtained results showed: the average limit of D-dimer was 0.8±0.5; prothrombin time 13.2±1.1 seconds; INR 0.98±0.08, prothrombin index 104.3±19.5%, EHS - 34.8±13.6 mm/s. It should be noted that respiratory distress syndrome (RDS), premature birth, malformed and extremely malformed newborns, asphyxia or hypoxia have been reported in infants born to pregnant women infected with the coronavirus disease.Thus, from the obtained indicators, it is known that pregnant women infected with the virus have a high risk of serious illness and death for both themselves and their babies. It has been proven that the majority of babies born to SARS-CoV-2 positive mothers have a negative impact on their health.

Keywords: Covid 19, reproductive health, preqnancy, premature birth

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Authors: Yasser Ahmed Mahmoud Ali Helal

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The use of CAD (Computer Aided Design) technology is ubiquitous in the architecture, engineering and construction (AEC) industry. This has led to its inclusion in the curriculum of architecture schools in Nigeria as an important part of the training module. This article examines the ethical issues involved in implementing CAD (Computer Aided Design) content into the architectural education curriculum. Using existing literature, this study begins with the benefits of integrating CAD into architectural education and the responsibilities of different stakeholders in the implementation process. It also examines issues related to the negative use of information technology and the perceived negative impact of CAD use on design creativity. Using a survey method, data from the architecture department of University was collected to serve as a case study on how the issues raised were being addressed. The article draws conclusions on what ensures successful ethical implementation. Millions of people around the world suffer from hepatitis C, one of the world's deadliest diseases. Interferon (IFN) is treatment options for patients with hepatitis C, but these treatments have their side effects. Our research focused on developing an oral small molecule drug that targets hepatitis C virus (HCV) proteins and has fewer side effects. Our current study aims to develop a drug based on a small molecule antiviral drug specific for the hepatitis C virus (HCV). Drug development using laboratory experiments is not only expensive, but also time-consuming to conduct these experiments. Instead, in this in silicon study, we used computational techniques to propose a specific antiviral drug for the protein domains of found in the hepatitis C virus. This study used homology modeling and abs initio modeling to generate the 3D structure of the proteins, then identifying pockets in the proteins. Acceptable lagans for pocket drugs have been developed using the de novo drug design method. Pocket geometry is taken into account when designing ligands. Among the various lagans generated, a new specific for each of the HCV protein domains has been proposed.

Keywords: drug design, anti-viral drug, in-silicon drug design, hepatitis C virus (HCV) CAD (Computer Aided Design), CAD education, education improvement, small-size contractor automatic pharmacy, PLC, control system, management system, communication

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Authors: Tessalyn Morrison, Alexis Dinno, Taurica Salmon

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Sex and gender conflation continue to perpetuate the invisibility of gender minorities and obscure information about the ways that biological sex and gender affect health. The misuse of sex and gender terms, and their respective binaries, can yield inaccurate results. But more importantly, it contributes to the erasure of sex and gender minority health experiences. This paper discusses ways in which public health researchers can use sex and gender terms correctly and center the health experiences of intersex, transgender, non binary, and a-gender individuals. It includes promoting sensitivity in approaching minority communities, improving survey questions, and collaborating with sex and gender minority communities to improve research quality and participant experiences. Improving our standards for the quality of sex and gender term usage and centering sex and gender minorities in public health research are imperative to address the health inequalities faced by sex and gender minorities.

Keywords: epidemiology, gender, intersex, research methods, sex, transgender

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Authors: Babanyara Y. Y., Ibrahim D. B., Garba T., Bogoro A. G., Abubakar, M. Y.

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Medical care is vital for our life, health, and well-being. However, the waste generated from medical activities can be hazardous, toxic, and even lethal because of their high potential for diseases transmission. The hazardous and toxic parts of waste from healthcare establishments comprising infectious, medical, and radioactive material as well as sharps constitute a grave risks to mankind and the environment, if these are not properly treated/disposed or are allowed to be mixed with other municipal waste. In Nigeria, practical information on this aspect is inadequate and research on the public health implications of poor management of medical wastes is few and limited in scope. Findings drawn from Literature particularly in the third world countries highlights financial problems, lack of awareness of risks involved in MWM, lack of appropriate legislation and lack of specialized MWM staff. The paper recommends how MWM practices can be improved in medical facilities.

Keywords: environmental pollution, infectious, management, medical waste, public health

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Authors: M. Kumpugdee-Vollrath, T. G. D. Phu, M. Helmis

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A suitable model membrane to study the pharmacological effect of pharmaceutical products is human stratum corneum because this layer of human skin is the outermost layer and it is an important barrier to be passed through. Other model membranes which were also used are for example skins from pig, mouse, reptile or fish. We are interested in fish skins in this project. The advantages of the fish skins are, that they can be obtained from the supermarket or fish shop. However, the fish skins should be freshly prepared and used directly without storage. In order to understand the effect of different model drugs e.g. lidocaine HCl, resveratrol, paracetamol, ibuprofen, acetyl salicylic acid on the properties of the model membrane from various types of fishes e.g. trout, salmon, cod, plaice permeation tests were performed and differential scanning calorimetry was applied.

Keywords: fish skin, model membrane, permeation, DSC, lidocaine HCl, resveratrol, paracetamol, ibuprofen, acetyl salicylic acid

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Authors: Abdelsabour G. A. Khaled, Ahmed W. A. Abdalla And Sabry Y. M. Mahmoud

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Banana plants showing typical mosaic and yellow stripes on leaves as symptoms were collected from Assiut Governorate in Egypt. The causal agent was identified as Cucumber mosaic virus (CMV) on the basis of symptoms, transmission, serology, transmission electron microscopy and reverse transcription polymerase chain reaction (RT-PCR). Coat protein (CP) gene was amplified using gene specific primers for coat protein (CP), followed by cloning into desired cloning vector for sequencing. In this study the CMV was transmitted into propagation host either by aphid or mechanically. The transmission was confirmed through Direct Antigen Coating Enzyme Linked Immuno Sorbent Assay (DAC-ELISA). Analysis of the 120 deduced amino acid sequence of the coat protein gene revealed that the EG-A strain of CMV shared from 97.50 to 98.33% with those strains belonging to subgroup IA. The cluster analysis grouped the Egyptian isolate with strains Fny and Ri8 belonging sub-group IA. It appears that there occurs a high incidence of CMV infecting banana belonging to IA subgroup in most parts of Egypt.

Keywords: banana, CMV, transmission, CP gene, RT-PCR

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Authors: Chijindu Okpalaoka, Charles Chinedu Onuselogu

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COVID-19, humanity's coronavirus disease caused by SARS-CoV-2, was first detected in late 2019 in Wuhan, China. COVID-19 lacked an established conventional pharmaceutical therapy, and as a result, the outbreak quickly became an epidemic affecting the entire World. Only a qPCR assay is reliable for diagnosing COVID-19. Clustered, regularly interspaced short palindromic repeats (CRISPR) technology is being researched for speedy and specific identification of COVID-19, among other therapeutic techniques. Apart from its therapeutic capabilities, the CRISPR technique is being evaluated to develop antiviral therapies; nevertheless, no CRISPR-based medication has been approved for human use to date. Prophylactic antiviral CRISPR in living being cells, a Cas 13-based approach against coronavirus, has been developed. While this method can be evolved into a treatment approach, it may face substantial obstacles in human clinical trials for licensure. This study discussed the potential applications of CRISPR-based techniques for developing a speedy and accurate feasible treatment alternative for the COVID-19 virus.

Keywords: COVID-19, CRISPR technique, Cas13, SARS-CoV-2, prophylactic antiviral

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Authors: Ekaterina Sánchez Romero, Emily Gabriela Aguirre Herrera, Sandra Luz Espinoza Esquerra, Jorge García Campos

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Female, 7 months old, daughter of a mother with anemia during pregnancy, with no history of atopy in the family, since birth she presents with recurrent dermatological and gastrointestinal infections, chronically treated for recurrent diaper dermatitis. At 6 months of age, she begins with generalized pallor, hyperpigmentation in hands and feet, smooth tongue, psychomotor retardation with lack of head support, sedation, and hypoactivity. She was referred to our hospital for a fever of 38°C, severe diaper rash, and pancytopenia with HB 9.3, platelets 38000, neutrophils 0.39 MCV: 86.80 high for her age. The approach was initiated to rule out myeloproliferative syndrome, with negative immunohistochemical results of bone marrow aspirate; during her stay, she presented neurological regression, lack of sucking, and focal seizures. CT scan showed cortical atrophy. The patient was diagnosed with primary immunodeficiency due to history; gamma globulin was administered without improvement with normal results of immunoglobulins and metabolic screening. When dermatological and neurological diagnoses were ruled out as the primary cause, a nutritional factor was evaluated, and a therapeutic trial was started with the administration of vitamin B12 and zinc, presenting clinical neurological improvement and resolution of pancytopenia in 2 months. It was decided to continue outpatient management. Discussion: We present a patient with neurological, dermatological involvement, and pancytopenia, so the most common differential diagnoses in this population were ruled out. Vitamin B12 deficiency is an uncommon entity. Due to maternal and clinical history, a therapeutic trial was started resulting in an improvement. Conclusion: VitaminB12 deficiency should be considered one of the differential diagnoses in the approach to pancytopenia with megaloblastic anemia associated with dermatologic and neurologic manifestations. Early treatment can reduce irreversible damage in these patients.

Keywords: vitamin B12 deficiency, pediatrics, pancytopenia, diaper dermatitis

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Authors: Ilyasu G., Ogoina D., Otu AA, Muhammed FD, Ebenso B., Otokpa D., Rotifa S., Tuduo-Wisdom O., Habib AG

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Background: The 2014 Ebola Virus Disease (EVD) outbreak was characterized by fear, misconceptions and irrational behaviors. We conducted a knowledge attitude and practice survey of EVD in Nigeria to inform the institution of effective control measures. Methods: Between July 30th and September 30th 2014, a cross-sectional study on knowledge, attitude and practice (KAP) of Ebola Virus Disease (EVD) was undertaken among adults of the general population and healthcare workers (HCW) in three states of Nigeria, including Kano, Cross River and Bayelsa states. Demographic information and data on KAP were obtained using a self-administered standardized questionnaire. The percentage KAP scores were categorized as good and poor. Independent predictors of good knowledge of EVD were ascertained using a binary logistic regression model. Results: Out of 1035 study participants with a median age of 32 years, 648 (62.6%) were males, 846 (81.7%) had tertiary education and 441 (42.6%) were HCW. There were 218, 239 and 578 respondents from Bayelsa, Cross Rivers, and Kano states, respectively. The overall median percentage KAP scores and interquartile ranges (IQR) were 79.46% (15.07%), 95.0% (33.33%), and 49.95% (37.50%), respectively. Out of the 1035 respondents, 470 (45.4%), 544(52.56%), and 252 (24.35%) had good KAP of EVD defined using 80%, 90%, and 70% score cut-offs, respectively. Independent predictors of good knowledge of EVD were a HCW (Odds Ratio-OR-2.89, 95% Confidence interval-CI of 1.41-5.90), reporting ‘moderate to high fear of EVD’ (OR-2.15, 95% CI-1.47-3.13) and ‘willingness to modify habit’ (OR-1.68, 95% CI-1.23-2.30). Conclusion: Our results reveal suboptimal EVD-related knowledge, attitude and practice among adults in Nigeria. To effectively control future outbreaks of EVD in Nigeria, there is a need to institute public sensitization programs that improve understanding of EVD and address EVD-related myths and misconceptions, especially among the general population.

Keywords: Ebola, health care worker, knowledge, attitude

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Authors: Mojtaba Zabihi, Stephen Munro, Jonathan Little, Ri Li, Joshua Brinkerhoff, Sina Kheirkhah

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Occupational Safety and Health Administration (OSHA) identified dental workers at the highest risk of contracting COVID-19. This is because aerosol-generating procedures (AGP) during dental practices generate aerosols ( < 5µm) and droplets. These particles travel at varying speeds, in varying directions, and for varying durations. If these particles bear infectious viruses, their spreading causes airborne transmission of the virus in the dental room, exposing dentists, hygienists, dental assistants, and even other dental clinic clients to the infection risk. Computational fluid dynamics (CFD) simulation of two-phase flows based on a discrete phase model (DPM) is carried out to study the spreading of aerosol and droplets in a dental room. The simulation includes momentum, heat, and mass transfers between the particles and the airflow. Two simulations are conducted and compared. One simulation focuses on the effects of room ventilation in winter and summer on the particles' travel. The other simulation focuses on the control of aerosol and droplets' spreading. A suction collector is added near the source of aerosol and droplets, creating a flow sink in order to remove the particles. The effects of the suction flow on the aerosol and droplet travel are studied. The suction flow can remove aerosols and also reduce the spreading of droplets.

Keywords: aerosols, computational fluid dynamics, COVID-19, dental, discrete phase model, droplets, two-phase flow

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Authors: Ugo Rovigatti

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Neuroblastoma (NBL) has epitomized for at least 40 years our understanding of cancer cellular and molecular biology and its potential applications to novel therapeutic strategies. This includes the discovery of the very first oncogene aberrations and tumorigenesis suppression by differentiation in the 80s; the potential role of suppressor genes in the 90s; the relevance of immunotherapy in the millennium first, and the discovery of additional mutations by NGS technology in the millennium second decade. Similar discoveries were achieved in the majority of human cancers, and similar therapeutic interventions were obtained subsequently to NBL discoveries. Unfortunately, targeted therapies suggested by specific mutations (such as MYCN amplification –MNA- present in ¼ or 1/5 of cases) have not elicited therapeutic successes in aggressive NBL, where the prognosis is still dismal. The reasons appear to be linked to Tumor Heterogeneity, which is particularly evident in NBL but also a clear hallmark of aggressive human cancers generally. The new avenue of cancer immunotherapy (CIT) provided new hopes for cancer patients, but we still ignore the cellular or molecular targets. CIT is emblematic of high-risk disease (HR-NBL) since the mentioned GD2 passive immunotherapy is still providing better survival. We recently critically reviewed and evaluated the literature depicting the genomic landscapes of HR-NBL, coming to the qualified conclusion that among hundreds of affected genes, potential targets, or chromosomal sites, none correlated with anti-GD2 sensitivity. A better explanation is provided by the Micro-Foci inducing Virus (MFV) model, which predicts that neuroblasts infection with the MFV, an RNA virus isolated from a cancer-cluster (space-time association) of HR-NBL cases, elicits the appearance of MNA and additional genomic aberrations with mechanisms resembling chromothripsis. Neuroblasts infected with low titers of MFV amplified MYCN up to 100 folds and became highly transformed and malignant, thus causing neuroblastoma in young rat pups of strains SD and Fisher-344 and larger tumor masses in nu/nu mice. An association was discovered with GD2 since this glycosphingolipid is also the receptor for the family of MFV virus (dsRNA viruses). It is concluded that a dsRNA virus, MFV, appears to provide better explicatory mechanisms for the genesis of i) specific genomic aberrations such as MNA; ii) extensive tumor heterogeneity and chromothripsis; iii) the effects of passive immunotherapy with anti-GD2 monoclonals and that this and similar models should be further investigated in both pediatric and adult cancers.

Keywords: neuroblastoma, MYCN, amplification, viruses, GD2

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Authors: Sabin Aslam, Sultan Habibullah Khan, James G. Thomson, Abhaya M. Dandekar

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Losses due to viral diseases are posing a serious threat to crop production. A quick breakdown of resistance to viruses like Cotton Leaf Curl Virus (CLCuV) demands the application of a proficient technology to engineer durable resistance. Gene stacking has recently emerged as a potential approach for integrating multiple genes in crop plants. In the present study, recombinase technology has been used for site-specific gene stacking. A target vector (pG-Rec) was designed for engineering a predetermined specific site in the plant genome whereby genes can be stacked repeatedly. Using Agrobacterium-mediated transformation, the pG-Rec was transformed into Coker-312 along with Nicotiana tabacum L. cv. Xanthi and Nicotiana benthamiana. The transgene analysis of target lines was conducted through junction PCR. The transgene positive target lines were used for further transformations to site-specifically stack two genes of interest using Bxb1 and PhiC31 recombinases. In the first instance, Cas9 driven by multiplex gRNAs (for Rep gene of CLCuV) was site-specifically integrated into the target lines and determined by the junction PCR and real-time PCR. The resulting plants were subsequently used to stack the second gene of interest (AVP3 gene from Arabidopsis for enhancing cotton plant growth). The addition of the genes is simultaneously achieved with the removal of marker genes for recycling with the next round of gene stacking. Consequently, transgenic marker-free plants were produced with two genes stacked at the specific site. These transgenic plants can be potential germplasm to introduce resistance against various strains of cotton leaf curl virus (CLCuV) and abiotic stresses. The results of the research demonstrate gene stacking in crop plants, a technology that can be used to introduce multiple genes sequentially at predefined genomic sites. The current climate change scenario highlights the use of such technologies so that gigantic environmental issues can be tackled by several traits in a single step. After evaluating virus resistance in the resulting plants, the lines can be a primer to initiate stacking of further genes in Cotton for other traits as well as molecular breeding with elite cotton lines.

Keywords: cotton, CRISPR/Cas9, gene stacking, genome editing, recombinases

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Authors: K. B. Chathyushya, M. Shiva Prakash, R. Hemalatha

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Introduction: Gastrointestinal (GI) tract has a number of microorganisms (microbiota) that influences the host’s health. The imbalance in the gut microbiota, which is also called as gut dysbiosis, affects human health which causes various metabolic, inflammatory, and infectious diseases. Probiotics play an important role in reinstating the gut balance. Probiotics are involved in the maintenance of healthier gut microbiota and have also been identified as effective adjuvants in insulin resistance therapies. Methods: This paper systematically reviews different randomized, controlled, blinded trials of probiotics for the treatment of various diseases along with the therapeutic or prophylactic properties of probiotic bacteria in different metabolic, inflammatory, infectious and anxiety-related disorders. Conclusion: The present review summarises that probiotics have some considerable effect in the management of various diseases, however, the benefits are strain specific, although more clinical trials are need to be carried out with different probiotic and symbiotic combinations as some probiotics have broad spectrum of benefits and few with specific activity

Keywords: life style diseases, cognition, health, gut dysbiosis, probiotics

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Authors: P.P. Rupasinghe, A.H. Farid

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The Aleutian mink disease virus (AMDV, Carnivore amdoparvovirus 1) causes persistent infection, plasmacytosis, and formation and deposition of immune complexes in various organs in adult mink, leading to glomerulonephritis, arteritis and sometimes death. The disease has no cure nor an effective vaccine, and identification and culling of mink positive for anti-AMDV antibodies have not been successful in controlling the infection in many countries. The failure to eradicate the virus from infected farms may be caused by keeping false-negative individuals on the farm, virus transmission from wild animals, or neighboring farms. The identification of sources of infection, which can be performed by comparing viral sequences, is important in the success of viral eradication programs. High mutation rates could cause inaccuracies when viral sequences are used to trace back an infection to its origin. There is no published information on the mutation rate of AMDV either in vivo or in vitro. The in vivo estimation is the most accurate method, but it is difficult to perform because of the inherent technical complexities, namely infecting live animals, the unknown numbers of viral generations (i.e., infection cycles), the removal of deleterious mutations over time and genetic drift. The objective of this study was to determine the mutation rate of AMDV on which no information was available. A homogenate was prepared from the spleen of one naturally infected American mink (Neovison vison) from Nova Scotia, Canada (parental template). The near full-length genome of this isolate (91.6%, 4,143 bp) was bidirectionally sequenced. A group of black mink was inoculated with this homogenate (descendant mink). Spleen sampled were collected from 10 descendant mink after 16 weeks post-inoculation (wpi) and from anther 10 mink after 176 wpi, and their near-full length genomes were bi-directionally sequenced. Sequences of these mink were compared with each other and with the sequence of the parental template. The number of nucleotide substitutions at 176 wpi was 3.1 times greater than that at 16 wpi (113 vs 36) whereas the estimates of mutation rate at 176 wpi was 3.1 times lower than that at 176 wpi (2.85×10-3 vs 9.13×10-4 substitutions/ site/ year), showing a decreasing trend in the mutation rate per unit of time. Although there is no report on in vivo estimate of the mutation rate of DNA viruses in animals using the same method which was used in the current study, these estimates are at the higher range of reported values for DNA viruses determined by various techniques. These high estimates are logical based on the wide range of diversity and pathogenicity of AMDV isolates. The results suggest that increases in the number of nucleotide substitutions over time and subsequent divergence make it difficult to accurately trace back AMDV isolates to their origin when several years elapsed between the two samplings.

Keywords: Aleutian mink disease virus, American mink, mutation rate, nucleotide substitution

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