Search results for: herpes simplex virus type I ELISA

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: Wei Wen Yang, Emily Chia Yu Su

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Dengue fever is a mosquito-borne tropical infectious disease caused by the dengue virus. After infection, symptoms usually start from three to fourteen days. Dengue virus may cause a high fever and at least two of the following symptoms, severe headache, severe eye pain, joint pains, muscle or bone pain, vomiting, feature skin rash, and mild bleeding manifestation. In addition, recovery will take at least two to seven days. Dengue fever has rapidly spread in tropical and subtropical areas in recent years. Several phenomena around the world such as global warming, urbanization, and international travel are the main reasons in boosting the spread of dengue. In Taiwan, epidemics occur annually, especially during summer and fall seasons. On the other side, Singapore government also has announced the amounts number of dengue cases spreading in Singapore. As the serious epidemic of dengue fever outbreaks in Taiwan and Singapore, countries around the Asia-Pacific region are becoming high risks of susceptible to the outbreaks and local hub of spreading the virus. To improve public safety and public health issues, firstly, we are going to use Microsoft Excel and SAS EG to do data preprocessing. Secondly, using support vector machines and decision trees builds predict model, and analyzes the infectious cases between Taiwan and Singapore. By comparing different factors causing vector mosquito from model classification and regression, we can find similar spreading patterns where the disease occurred most frequently. The result can provide sufficient information to predict the future dengue infection outbreaks and control the diffusion of dengue fever among countries.

Keywords: dengue fever, Taiwan, Singapore, Aedes aegypti

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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: Nazanin Ahmadi Daryakenari

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Background: Diabetes significantly influences the risk of heart failure. The interplay between distinct types of diabetes, heart failure, and their distribution across various age groups remains an area of active exploration. This study endeavors to scrutinize the age group distribution in publications addressing Type 1 and Type 2 diabetes and heart failure on PubMed while also examining the evolving publication trends. Methods: We leveraged E-utilities and RegEx to search and extract publication data from PubMed using various mesh terms. Subsequently, we conducted descriptive statistics and t-tests to discern the differences between the two diabetes types and the distribution across age groups. Finally, we analyzed the temporal trends of publications concerning both types of diabetes and heart failure. Results: Our findings revealed a divergence in the age group distribution between Type 1 and Type 2 diabetes within heart failure publications. Publications discussing Type 2 diabetes and heart failure were more predominant among older age groups, whereas those addressing Type 1 diabetes and heart failure displayed a more balanced distribution across all age groups. The t-test revealed no significant difference in the means between the two diabetes types. However, the number of publications exploring the relationship between Type 2 diabetes and heart failure has seen a steady increase over time, suggesting an escalating interest in this area. Conclusion: The dissection of publication patterns on PubMed uncovers a pronounced association between Type 2 diabetes and heart failure within older age groups. This highlights the critical need to comprehend the distinct age group differences when examining diabetes and heart failure to inform and refine targeted prevention and treatment strategies.

Keywords: Type 1 diabetes, Type 2 diabetes, heart failure, age groups, publication patterns, PubMed

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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: A. M. Abd-Elfattah, M. H. Abu-Moussa

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In this paper, the estimation of stress-strength parameter R = P(Y < X) is considered when X; Y the strength and stress respectively are two independent random variables of Burr Type XII distribution. The samples taken for X and Y are progressively censoring of type II. The maximum likelihood estimator (MLE) of R is obtained when the common parameter is unknown. But when the common parameter is known the MLE, uniformly minimum variance unbiased estimator (UMVUE) and the Bayes estimator of R = P(Y < X) are obtained. The exact condence interval of R based on MLE is obtained. The performance of the proposed estimators is compared using the computer simulation.

Keywords: Burr Type XII distribution, progressive type-II censoring, stress-strength model, unbiased estimator, maximum-likelihood estimator, uniformly minimum variance unbiased estimator, confidence intervals, Bayes estimator

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Authors: Lavrentiadou S. N., Angelou V., Chatzimisios K., Papazoglou L.

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The aim of this study was the isolation and culture of keratinocytes and fibroblasts from feline skin to ultimately create an artificial engineered skin (including dermis and epidermis) useful for the effective treatment of large cutaneous deficits in cats. Epidermal keratinocytes and dermal fibroblasts were freshly isolated from skin biopsies using an 8 mm biopsy punch obtained from 8 healthy cats that had undergone ovariohysterectomy. The owner’s consent was obtained. All cats had a complete blood count and a serum biochemical analysis and were screened for feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV) preoperatively. The samples were cut into small pieces and incubated with collagenase (2 mg/ml) for 5-6 hours. Following digestion, cutaneous cells were filtered through a 100 μm cell strainer, washed with DMEM, and grown in DMEM supplemented with 10% FBS. The undigested epidermis was washed with DMEM and incubated with 0.05% Trypsin/0.02% EDTA (TE) solution. Keratinocytes recovered in the TE solution were filtered through a 100 μm and a 40 μm cell strainer and, following washing, were grown on a collagen type I matrix in DMEM: F12 (3:1) medium supplemented with 10% FΒS, 1 μm hydrocortisone, 1 μm isoproterenol and 0.1 μm insulin. Both fibroblasts and keratinocytes were grown in a humidified atmosphere with 5% CO2 at 37oC. The medium was changed twice a week and cells were cultured up to passage 4. Cells were grown to 70-85% confluency, at which point they were trypsinized and subcultured in a 1:4 dilution. The majority of the cells in each passage were transferred to a freezing medium and stored at -80oC. Fibroblasts were frozen in DMEM supplemented with 30% FBS and 10% DMSO, whereas keratinocytes were frozen in a complete keratinocyte growth medium supplemented with 10% DMSO. Both cell types were thawed and successfully grown as described above. Therefore, we can create a bank of fibroblasts and keratinocytes, from which we can recover cells for further culture and use for the generation of skin equivalent in vitro. In conclusion, cutaneous cell isolation and cell culture and expansion were successfully developed. To the authors’ best knowledge, this is the first study reporting isolation and culture of keratinocytes and fibroblasts from feline skin. However, these are preliminary results and thus, the development of autologous-engineered feline skin is still in process.

Keywords: cat, fibroblasts, keratinocytes, skin equivalent, wound

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Authors: Sumeyra Sevim, Gulsum Gizem Topal, Mercan Merve Tengilimoglu-Metin, Mevlude Kizil

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Aflatoxin M1 (AFM1) is a major toxic and carcinogenic molecule in milk and milk products. Therefore, it poses a risk for public health. Probiotics can be biological agent to remove AFM1. The aim of this study is to evaluate the effect of probiotic bacteria on AFM1 detoxification in phosphate buffer saline. The PBS samples artificially contaminated with AFM1 at concentration 100 pg/ml were prepared with probiotics bacteria that including monoculture (L. plantarum, B. bifidum ATCC, B. animalis ATCC 27672) and binary culture (L. bulgaricus + S. thermophiles, B. bifidum ATCC + B. animalis ATCC 27672, L. plantarum+B. bifidum ATCC, L. plantarum+ B. animalis ATCC 27672). The samples were incubated at 37°C for 4 hours and stored for 1, 5 and 10 days. The toxin was measured by the ELISA. The highest levels of AFM1 binding ability (63.6%) in PBS were detected yoghurt starter bacteria, while L. plantarum had the lowest levels of AFM1 binding ability (35.5%) in PBS. In addition, it was found that there was significant effect of storage on AFM1 binding ability in all groups except the one including B. animalis (p < 0.05). Consequently, results demonstrate that AFM1 detoxification by probiotic bacteria has a potential application to reduce toxin concentrations in yoghurt. Moreover, probiotic strains can react with itself as synergic or antagonist.

Keywords: aflatoxin M1, ELISA, probiotics, storage

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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: Fengmei Li, Li Xu, Guoliang Xia

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Lysozyme is a catalytic enzyme that performs bacterial cell lysis by cleaving the β-1,4-glycosidic bond between N-acetylmuramic acid and N-acetylglucosamine of peptidoglycan in cell walls. In the present study, an invertebrate-type (i-type) lysozyme gene was cloned from Chinese mitten crab Eriocheir sinensis (designated as EsLysozyme) based on PCR-based rapid amplification of cDNA ends (RACE) technology. The full-length cDNA of EsLysozyme was of 831 bp. SMART and SIGNALP 3.0 program analysis revealed that EsLysozyme contained a signal peptide and a destabilase domain. The five amino acid residues (Tyr63, Trp64, Tyr91, His110, Pro114) and the conserved motif GSLSCG(P/Y)FQI and CL(E/L/R/H)C(I/M)C in i-type lysozymes were also found in EsLysozyme. The high similarity of EsLysozyme with L. vannamei lysozymes and phylogenetic analysis suggested that EsLysozyme should be a new member of i-type lysozyme family.

Keywords: i-type lysozyme, Eriocheir sinensis, cloning, characterization

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Authors: Ouahiba Bechiri, Mostefa Abbessi

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In recent year, polyoxometallates are intensely being explored because of their applications as new materiels, structural aesthetics, catalysts, and biologically active compounds. heteropolyanions of general formulae [X2M18O62] n- (X= heteroatom, e.g. P, Si) and (M=W, Mo), known as Dawson-type anions, constitue a special class of polyoxometallate compounds. In this present work, cobalt substituted heteropolyanion Dawson-type [HP2W15Mo3CoO61] were synthesized and characterized by IR spectroscopy, 31 P NMR, cyclic voltammetry.

Keywords: heteropolyanions, nanomaterials, Dawson-type, characterization

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Authors: Kathryn H. Yull, Lina T. Al Kury, Frank Christopher Howarth

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Cardiovascular diseases are frequently reported in patients with Type-1 Diabetes mellitus (DM). In addition to changes in cardiac muscle inotropy, electrical abnormalities are also commonly observed in these patients. In the present study, using streptozotocin (STZ) rat model of Type-1 DM, we have characterized the changes in L-type calcium channel activity in single atrioventricular nodal (AVN) myocytes. Ionic currents were recorded from AVN myocytes isolated from the hearts of control rats and from those with STZ-induced diabetes. Patch-clamp recordings were used to assess changes in cellular electrical activity in individual myocytes. Type-1 DM significantly altered the cellular characteristics of L-type calcium current (ICaL). A reduction in peak ICaL density was observed, with no corresponding changes in the activation parameters of the current. ICaL also exhibited faster time-dependent inactivation in AVN myocytes from diabetic rats. A negative shift in the voltage dependence of inactivation was also evident. These findings demonstrate that experimentally–induced type-1 DM significantly alters AVN L-type calcium channel cellular electrophysiology. The changes in ion channel activity may underlie the abnormalities in the cardiac electrical function that contribute to the high mortality levels in patients with DM.

Keywords: cardiac, ion-channel, diabetes, atrioventricular node, calcium channel

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Authors: Ferreira RC, Simons HZ, Thompson WS, Cutler AJ, Dopico XC, Smyth DJ, Mashar M, Schuilenburg H, Walker NM, Dunger DB, Wallace C, Todd JA, Wicker LS, Pekalski ML

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Type 1 diabetes is caused by autoimmune destruction of insulin-secreting beta cells in the pancreas. T cells are known to play an important role in this immune-mediated destruction; however, there is no general consensus regarding alterations in cytokine production or T cell subsets in peripheral blood of patients with type 1 diabetes. Using polychromatic flow cytometry of peripheral blood mononuclear cells (PBMCs), we assessed production of the proinflammatory cytokines IL-21, IFN-γ and IL-17 by memory CD4 T effector (Teff) cells in 69 patients with type 1 diabetes and 61 healthy donors. We found a 21.9% (95% CI 5.8, 40.2; p = 3.9 × 10(-3)) higher frequency of IL-21(+) CD45RA(-) memory CD4(+) Teffs in patients with type 1 diabetes (geometric mean 5.92% [95% CI 5.44, 6.44]) compared with healthy donors (geometric mean 4.88% [95% CI 4.33, 5.50]). In a separate cohort of 30 patients with type 1 diabetes and 32 healthy donors, we assessed the frequency of circulating T follicular helper (Tfh) cells in whole blood. Consistent with the increased production of IL-21, we also found a 14.9% increase in circulating Tfh cells in the patients with type 1 diabetes (95% CI 2.9, 26.9; p = 0.016). Analysis of IL-21 production by PBMCs from a subset of 46 of the 62 donors immunophenotyped for Tfh showed that frequency of Tfh cells was associated with the frequency of IL-21+ cells (r2 = 0.174, p = 0.004). These results indicate that increased IL-21 production is likely to be an aetiological factor in the pathogenesis of type 1 diabetes that could be considered as a potential therapeutic target.

Keywords: T follicular helper cell, IL-21, IL-17, type 1 diabetes

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Authors: Jaeyoung Kim, Heeju Lee, Huijin Jung, Heesoo Pyo, Seungki Kim, Joonseok Lee

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Avian influenza viruses (AIV) are the primary cause of highly contagious respiratory diseases caused by type A influenza viruses of the Orthomyxoviridae family. AIV are categorized on the basis of types of surface glycoproteins such as hemagglutinin and neuraminidase. Certain H5 and H7 subtypes of AIV have evolved to the high pathogenic avian influenza (HPAI) virus, which has caused considerable economic loss to the poultry industry and led to severe public health crisis. Several commercial kits have been developed for on-site detection of AIV. However, the sensitivity of these methods is too low to detect low virus concentrations in clinical samples and opaque stool samples. Here, we introduced a background-free near-infrared (NIR)-to-NIR upconversion nanoparticle-based lateral flow immunoassay (NNLFA) platform to yield a sensor that detects AIV within 20 minutes. Ca²⁺ ion in the shell was used to enhance the NIR-to-NIR upconversion photoluminescence (PL) emission as a heterogeneous dopant without inducing significant changes in the morphology and size of the UCNPs. In a mixture of opaque stool samples and gold nanoparticles (GNPs), which are components of commercial AIV LFA, the background signal of the stool samples mask the absorption peak of GNPs. However, UCNPs dispersed in the stool samples still show strong emission centered at 800 nm when excited at 980 nm, which enables the NNLFA platform to detect 10-times lower viral load than a commercial GNP-based AIV LFA. The detection limit of NNLFA for low pathogenic avian influenza (LPAI) H5N2 and HPAI H5N6 viruses was 10² EID₅₀/mL and 10³.⁵ EID₅₀/mL, respectively. Moreover, when opaque brown-colored samples were used as the target analytes, strong NIR emission signal from the test line in NNLFA confirmed the presence of AIV, whereas commercial AIV LFA detected AIV with difficulty. Therefore, we propose that this rapid and background-free NNLFA platform has the potential of detecting AIV in the field, which could effectively prevent the spread of these viruses at an early stage.

Keywords: avian influenza viruses, lateral flow immunoassay on-site detection, upconversion nanoparticles

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Authors: Kriti Upadhyay, Ashraf Ali, Puja Sohal, Randeep Guleria

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Background: In Chronic Obstructive Pulmonary diseases (COPD) pathogenesis oxidative stress plays an important role. Hypoxia-Inducible factor (HIF-1α) is a dimeric protein complex which Functions as a master transcriptional regulator of the adaptive response to hypoxiaand is a risk factor that increases when oxidative stress triggers. The role ofHIF-1αin COPD due to smoking is lacking. Aim: This study aims to evaluate the role of HIF-1α in smoker COPD patients comparing its association with diseases severity. Method: In this cross-sectional study, we recruited 87 subjects, 57 were smokers with COPD,15 were smokers without COPD and other 15 were non-smoker healthy controls. The mean age was 54.6± 9.32 (cases 57.08±8.15; controls 50.0± 9.8). There were 62%smokers, 25% non-smokers,7% tobacco chewers and 6% ex-smokers. Enzyme-linked immune sorbent assay (ELISA) method was used for analyzing serum samples wherein HIF-1α was analyzed by Sandwich-ELISA. Results: In smoker COPD patients, a significantly higher HIF-1α level showed positive association with hypoxia, smoking status and severity of disease (p=0.03). The mean value of HIF-1α was not significantly different in smokers without COPD and healthy controls. Conclusion: It is found that HIF-1α level was increased in smoker COPD, but not in smokers without COPD. This suggests that development of COPD drive the HIF-1α pathway and it correlates with the severity of diseases.

Keywords: COPD, chronic obstructive pulmonary diseases, smokers, nonsmokers, hypoxia

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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: Abdusalam Gsiea, Ramadan Al-habashi, Mohamed Atumi, Khaled Atmimi

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We present a theoretical investigation on the structural, electronic properties and vibrational mode of nitrogen impurities in ZnO. The atomic structures, formation and transition energies and vibrational modes of (NO3)i interstitial or NO4 substituting on an oxygen site ZnO were computed using ab initio total energy methods. Based on Local density functional theory, our calculations are in agreement with one interpretation of bound-excition photoluminescence for N-doped ZnO. First-principles calculations show that (NO3)i defects interstitial or NO4 substituting on an Oxygen site in ZnO are important suitable impurity for p-type doping in ZnO. However, many experimental efforts have not resulted in reproducible p-type material with N2 and N2O doping. by means of first-principle pseudo-potential calculation we find that the use of NO or NO2 with O gas might help the experimental research to resolve the challenge of achieving p-type ZnO.

Keywords: DFF, nitrogen, p-type, ZnO

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Authors: Nouf A. Aldarmahi, Nesrin I. Tarbiah, Nuha A. Alkhattabi, Huda F. Alshaibi

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Nigella sativa which is known as dark cumin is a well-known example for a widely applicable herbal medicine. Nigella sativa can be effective in a variety of diseases such as hypertension, diabetes, bronchitis, gastrointestinal upset, and cancer. The anticancer effect of Nigella sativa appeared to be mediated by immune-modulatory effect through stimulating human natural killer (NK) cells. This is a type of lymphocytes which is part of the innate immunity, also known as the first line of defense in the body against pathogens. This study investigated the effect of thymoquinone as a major component of Nigella sativa on the molecular cytotoxic pathway of NK cell and the role of thymoquinone therapeutic effect on NK cells. NK cells were cultured with breast tumor cells in different ways and cultured media was collected and the concentration of perforin, granzyme B and interferon-α were measured by ELISA. The cytotoxic effect of NK cells on breast tumor cells was enhanced in the presence of thymoquinone, with increased activity of perforin in NK cells. This improved anticancer effect of thymoquinone on breast cancer cells.

Keywords: breast cancer, cancer cells, natural killer cells, thymoquinone

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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: 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: B. Falah, N. E. Abakouy

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Due to its high computational cost, mutation testing has been neglected by researchers. Recently, many cost and mutants’ reduction techniques have been developed, improved, and experimented, but few of them has relied the possibility of reducing the cost of mutation testing on the program type of the application under test. This paper is a comparative study between four operators’ selection techniques (mutants sampling, class level operators, method level operators, and all operators’ selection) based on the program code type of each application under test. It aims at finding an alternative approach to reveal the effect of code type on mutation testing score. The result of our experiment shows that the program code type can affect the mutation score and that the programs using polymorphism are best suited to be tested with mutation testing.

Keywords: equivalent mutant, killed mutant, mutation score, mutation testing, program code type, software testing

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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: Samul Sahile Kebede

Abstract:

Back ground: -Immune hemolytic anemia commonly affects human immune deficiency, infected individuals. Among anemic HIV patients in Africa, the burden of IHA due to autoantibody was ranged from 2.34 to 3.06 due to the drug was 43.4%. IHA due to autoimmune is potentially a fatal complication of HIV, which accompanies the greatest percent from acquired hemolytic anemia. Objective: -The main aim of this study was to determine the magnitude and associated factors of immune hemolytic anemia among human immuno deficiency virus infected adults at the university of Gondar comprehensive specialized hospital north west Ethiopia from March to April 2021. Methods: - An institution-based cross-sectional study was conducted on 358 human immunodeficiency virus-infected adults selected by systematic random sampling at the University of Gondar comprehensive specialized hospital from March to April 2021. Data for socio-demography, dietary and clinical data were collected by structured pretested questionnaire. Five ml of venous blood was drawn from each participant and analyzed by Unicel DHX 800 hematology analyzer, blood film examination, and antihuman globulin test were performed to the diagnosis of immune hemolytic anemia. Data was entered into Epidata version 4.6 and analyzed by STATA version 14. Descriptive statistics were computed and firth penalized logistic regression was used to identify predictors. P value less than 0.005 interpreted as significant. Result; - The overall prevalence of immune hemolytic anemia was 2.8 % (10 of 358 participants). Of these, 5 were males, and 7 were in the 31 to 50 year age group. Among individuals with immune hemolytic anemia, 40 % mild and 60 % moderate anemia. The factors that showed association were family history of anemia (AOR 8.30 at 95% CI 1.56, 44.12), not eating meat (AOR 7.39 at 95% CI 1.25, 45.0), and high viral load 6.94 at 95% CI (1.13, 42.6). Conclusion and recommendation; Immune hemolytic anemia is less frequent condition in human immunodeficiency virus infected adults, and moderate anemia was common in this population. The prevalence was increased with a high viral load, a family history of anemia, and not eating meat. In these patients, early detection and treatment of immune hemolytic anemia is necessary.

Keywords: anemia, hemolytic, immune, auto immune, HIV/AIDS

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Authors: R. Osada, S. Ogata, T. Segawa

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Dielectric barrier discharge plasma actuators (DBD-PAs) have been developed for active flow control devices. However, it is necessary to reduce ozone produced by DBD toward practical applications using DBD-PAs. In this study, variations of ozone concentration, flow velocity, power consumption were investigated by changing exposed electrodes of DBD-PAs. Two exposed electrode prototypes were prepared: span-type with exposed electrode width of 0.1 mm, and normal-type with width of 5 mm. It was found that span-type shows lower power consumption and higher flow velocity than that of normal-type at V_p-p = 4.0-6.0 kV. Ozone concentration of span-type higher than normal-type at V_p-p = 4.0-8.0 kV. In addition, it was confirmed that catalyst located in downstream from the exposed electrode can reduce ozone concentration between 18 and 42% without affecting the induced flow.

Keywords: dielectric barrier discharge plasma actuators, ozone diffusion, PIV measurement, power consumption

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Authors: Y. A. Yahaya, Ahmad Tijjani Asabe

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This paper focuses on the formulation of 3-step hybrid Adams type method for the solution of first order differential equation (ODE). The methods which was derived on both grid and off grid points using multistep collocation schemes and also evaluated at some points to produced Block Adams type method and Adams moulton method respectively. The method with the highest order was selected to serve as the corrector. The convergence was valid and efficient. The numerical experiments were carried out and reveal that hybrid Adams type methods performed better than the conventional Adams moulton method.

Keywords: adam-moulton type (amt), corrector method, off-grid, block method, convergence analysis

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Authors: Tejinder Singh

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Information system is the flow of data from different levels to different directions for decision making and data operations in information system (IS). Data can be violated by different manner like manual or technical errors, data tampering or loss of integrity. Security system called firewall of IS is effected by such type of violations. The flow of data among various levels of Information System is done by networking system. The flow of data on network is in form of packets or frames. To protect these packets from unauthorized access, virus attacks, and to maintain the integrity level, network security is an important factor. To protect the data to get pirated, various security techniques are used. This paper represents the various security techniques and signifies different harmful attacks with the help of detailed data analysis. This paper will be beneficial for the organizations to make the system more secure, effective, and beneficial for future decisions making.

Keywords: information systems, data integrity, TCP/IP network, vulnerability, decision, data

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

Abstract:

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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