Search results for: metamorphic virus

Authors: Bhawana, Roshan Kamal Topno, Maneesh Kumar, Major Madhukar, Krishna Pandey, Ganesh Chandra Sahoo, Manas Ranjan Dikhit, Surya Suman, Devendra Prasad Yadav, Rishikesh Kumar, Pradeep Das

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Viral co-infection is now very common across taxa and environments that are involved in congenital infections. Herpes simplex virus (HSV) and Rubella are the two serious viral infections, well categorized in TORCH Syndrome. Here we had endeavoured the seroprevalence of co-infection of HSV and Rubella. Systematic tests have been performed to check the virulence pattern of the co-infection. The study was conducted at Department of Virology, Rajendra Memorial Research Institute of Medical Sciences (ICMR), Patna, Bihar, India during January 2018-July 2018. 299 newly cases were attended with the sign and symptoms of HSV and Rubella. After taking written consent forms from all the subjects, blood samples were collected for serological detection. ELISA was performed to detect the presence of IgM antibody level. 12 patients were found to be IgM positive from each HSV and Rubella infection. The findings of our study showed that 6 patients were positive for both HSV and rubella and hence were co-infected. Such co-infection causes severe health problems as it leads to the mortality rate of the patients during viral infectivity. Epidemiologically, proper screening should be needed to check any chance of occurrence of such co-infection in the affected regions in large scale and take suitable preventive approach to decrease the case totality. Concern has to be given to aid proper diagnosis and treatment in order to decrease the spread of HSV and Rubella co-infection.

Keywords: HSV, Rubella, seroprevalence, co-infection, ELISA, viral infectivity

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Authors: Yik Sin Chan, Bee Ling Chuah, Wei Quan Chan, Ri Jin Cheng, Yan Hang Oon, Kong Soo Khoo, Nam Weng Sit

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Traditionally, medicinal plants have been used to treat different kinds of ailments including infectious diseases. They serve as a good source of lead compounds for the development of new and safer anti-infective agents. This study aimed to investigate the antimicrobial potential of the leaves of three medicinal plants, namely Catunaregam spinosa (Rubiaceae; Mountain pomegranate), Houttuynia cordata (Saururaceae; "fishy-smell herb") and Rhapis excelsa (Arecaceae; “broadleaf lady palm”). The leaves extracts were obtained by sequential extraction using hexane, chloroform, ethyl acetate, ethanol, methanol and water. The antibacterial and antifungal activities were assessed using a colorimetric broth microdilution method against a panel of human pathogenic bacteria (Gram-positive: Bacillus cereus and Staphylococcus aureus; Gram-negative: Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa) and fungi (yeasts: Candida albicans, Candida parapsilosis and Cryptococcus neoformans; Moulds: Aspergillus fumigatus and Trichophyton mentagrophytes) respectively; while antiviral activity was evaluated against the Chikungunya virus on monkey kidney epithelial (Vero) cells by neutral red uptake assay. All the plant extracts showed bacteriostatic activity, however, only 72% of the extracts (13/18) were found to have bactericidal activity. The lowest minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were given by the hexane extract of C. spinosa against S. aureus with the values of 0.16 and 0.31 mg/mL respectively. All the extracts also possessed fungistatic activity. Only the hexane, chloroform and ethyl acetate extracts of H. cordata exerted inhibitory activity against A. fumigatus, giving the lowest fungal susceptibility index of 16.7%. In contrast, only 61% of the extracts (11/18) showed fungicidal activity. The ethanol extract of R. excelsa exhibited the strongest fungicidal activity against C. albicans, C. parapsilosis and T. mentagrophytes with minimum fungicidal concentration (MFC) values of 0.04–0.08 mg/mL, in addition to its methanol extract against T. mentagrophytes (MFC=0.02 mg/mL). For anti-Chikungunya virus activity, only chloroform and ethyl acetate extracts of R. excelsa showed significant antiviral activity with 50% effective concentrations (EC50) of 29.9 and 78.1 g/mL respectively. Extracts of R. excelsa warrant further investigations into their active principles responsible for antifungal and antiviral properties.

Keywords: bactericidal, Chikungunya virus, extraction, fungicidal

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Authors: Nisha Thapa, Ram Prasad Mainali, Prakriti Chand

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Broadleaf mustard is a commercialized leafy vegetable of Nepal. However, its utilization is hindered in terms of production and productivity due to the high intensity of insects, pests, and diseases causing great loss. The plant protection part of the crop’s disease and damage intensity has not been studied much from research perspectives in Nepal. The research aimed to evaluate broadleaf mustard genotypes for resistance against different diseases. A total of 35 native genotypes of broadleaf mustard were screened at weekly intervals by scoring the plants for ten weeks. Five different diseases, such as Rhizoctonia root rot, Alternaria blight, black rot, turnip mosaic virus disease, and white rust, were reported from the broad leaf mustard genotypes. Out of 35 genotypes, 23 genotypes were found with very high Rhizoctonia Root Rot severity, whereas 8 genotypes showed very high Alternaria blight severity. Likewise, 3 genotypes were found with high Black rot severity, and 1 genotype was found with very high Turnip mosaic virus disease incidence. Similarly, 2 genotypes were found to have very high White rust severity. Among the disease of national importance, Rhizoctonia root rot was found to be the most severe disease with the greatest loss. Broadleaf mustard genotypes like Rato Rayo, CO 1002, and CO 11007 showed average to the high level of field resistance; therefore, these genotypes should be used, conserved, and stored in a mustard improvement program as the disease resistance quality or susceptibility of these genotypes can be helpful for seed producing farmers, companies and other stakeholders through varietal improvement and developmental works that further aids in sustainable disease management of the vegetable.

Keywords: genotype, disease resistance, Rhizoctonia root rot severity, varietal improvement

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Authors: Isomar Lima da Silva, Aristeu Jonatas Leite de Oliveira, Roberto Maia Augusto

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Biological isolation capsules are equipment commonly used in the control and prevention of infectious diseases in the hospital environment. This type of equipment, combined with pre-established medical protocols, contributes significantly to the containment of highly transmissible pathogens such as COVID-19. Due to its hermetic isolation, it allows more excellent patient safety, protecting companions and the health team. In this context, this work presents the development, testing, and validation of a medical capsule to treat patients affected by COVID-19. To this end, requirements such as low cost and easy handling were considered to meet the demand of people infected with the virus in remote locations in the Amazon region and/or where there are no ICU beds and mechanical ventilators for orotracheal intubation. Conceived and developed in a partnership between SAMEL Planos de Saúde and Instituto Conecthus, the device entitled "Vanessa Capsule" was designed to be used together with the NIV protocol (non-invasive ventilation), has an automatic exhaust system and filters performing the CO2 exchange, in addition to having BiPaps ventilatory support equipment (mechanical fans) in the Cabin Kit. The results show that the degree of effectiveness in protecting against infection by aerosols, with the protection cabin, is satisfactory, implying the consideration of the Vanessa capsule as an auxiliary method to be evaluated by the health team. It should also be noted that the medical observation of the evaluated patients found that the treatment against the COVID-19 virus started earlier with non-invasive mechanical ventilation reduces the patient's suffering and contributes positively to their recovery, in association with isolation through the Vanessa capsule.

Keywords: COVID-19, mechanical ventilators, medical capsule, non-invasive ventilation

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Authors: T. Ferreira, A. Kulkarni, C. Bretscher, K. Richter, M. Ehrlich, A. Marchini

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H-1 protoparvovirus (H-1PV) is a virus with inherent oncolytic and oncosuppressive activities while remaining non-pathogenic in humans. H-1PV was the first oncolytic parvovirus to undergo clinical testing. Results from trials in patients with glioblastoma or pancreatic carcinoma showed an excellent safety profile and first signs of efficacy. H-1PV infection is vastly dependent on cellular factors, from cell attachment and entry to viral replication and egress. Hence, we believe that the characterisation of the parvovirus life cycle would ultimately help further improve H-1PV clinical outcome. In the present study, we explored the entry pathway of H-1PV in cervical HeLa and glioma NCH125 cancer cell lines. Electron and confocal microscopy showed viral particles associated with clathrin-coated pits and vesicles, providing the first evidence that H-1PV cell entry occurs through clathrin-mediated endocytosis. Accordingly, we observed that by blocking clathrin-mediated endocytosis with hypertonic sucrose, chlorpromazine, or pitstop 2, H-1PV transduction was markedly decreased. Accordingly, siRNA-mediated knockdown of AP2M1, which retains a crucial role in clathrin-mediated endocytosis, verified the reliance of H-1PV on this route to enter HeLa and NCH125 cancer cells. By contrast, we found no evidence of viral entry through caveolae-mediated endocytosis. Indeed, pre-treatment of cells with nystatin or methyl-β-cyclodextrin, both inhibitors of caveolae-mediated endocytosis, did not affect viral transduction levels. Unexpectedly, siRNA-mediated knockdown of caveolin-1, the main driver of caveolae-mediated endocytosis, increased H-1PV transduction, suggesting caveolin-1 is a negative modulator of H-1PV infection. We also show that H-1PV entry is dependent on dynamin, a protein responsible for mediating the scission of vesicle neck and promoting further internalisation. Furthermore, since dynamin inhibition almost completely abolished H-1PV infection, makes it unlikely that H-1PV uses macropinocytosis as an alternative pathway to enter cells. After viral internalisation, H-1PV passes through early to late endosomes as observed by confocal microscopy. Inside these endocytic compartments, the acidic environment proved to be crucial for a productive infection. Inhibition of acidification of pH dramatically reduced H-1PV transduction. Besides, a fraction of H-1PV particles was observed inside LAMP1-positive lysosomes, most likely following a non-infectious route. To the author's best knowledge, this is the first study to characterise the cell entry pathways of H-1PV. Along these lines, this work will further contribute to understand H-1PV oncolytic properties as well as to improve its clinical potential in cancer virotherapy.

Keywords: clathrin-mediated endocytosis, H-1 parvovirus, oncolytic virus, virus entry

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Authors: JiYoung Park, SueGoo Rhee, HyunAe Woo

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In liver regeneration, quiescent hepatocytes need to be primed to fully respond to growth factors such as hepatocyte growth factor. To understand the priming process, it is necessary to analyze patterns of gene expression that occur during liver regeneration after partial hepatectomy (PHx). Recently, tyrosine phosphorylation of signal transducer and activator of transcription 3 (pYSTAT3) has been shown to play an important role in initiating liver regeneration. In order to evaluate the role of pYSTAT3 on liver regeneration after PHx, we used an intrabody which can selectively inhibit pYSTAT3. In our previous studies, an intrabody had been shown that it bound specifically to the pYSTAT3. Adenovirus-mediated expression of the intrabody in HepG2 cells, as well as mouse liver, blocked both accumulation of pYSTAT3 in the nucleus and downstream target of pYSTAT3. In this study, PHx was performed on intrabody-expressing mice and the expression levels of liver regeneration-related genes were analyzed. We also measured liver/body weight ratios and the related cellular signaling pathways were analyzed. Acute phase response genes were reduced in an intrabody-expressing mice during liver regeneration than in control virus-injected mice. However, the time course of liver mass restoration in intrabody-expressing mice was similar to that observed in control virus-injected mice. We also observed that the expression levels of anti-apoptotic genes, such as Bcl2 and Bcl-xL were decreased in intrabody-expressing mice whereas the expression of cell cycle-related genes such as cyclin D1, and c-myc was increased. Liver regeneration after PHx was partially impaired by the selective inhibition of pYSTAT3 with a phosphorylation site-specific intrabody and these results indicated that pYSTAT3 might have limited role in liver mass recovery.

Keywords: STAT3, pYSTAT3, liver regeneration, intrabody

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Authors: Sahar Heidary, Ramin Ghasemi Shayan

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The recent coronavirus disease 2019 (COVID-19) occurrence has carried considerabletrials to the world health system, comprising the training of dental and maxillofacial radiology (DMFR). DMFR will keep avital role in healthcare throughout this disaster. Severe acute breathing disease coronavirus 2 (SARS-CoV-2), the virus producing the current coronavirus disease 2019 (COVID-19) pandemic, is not only extremely contagious but can make solemn consequences in susceptible persons comprising dental patients and dental health care personnel (DHCPs). Reactions to COVID-19 have been available by the Cores for Infection Switch and Inhibition and the American Dental Association, but a more detailed answer is necessary for the harmless preparation of oral and maxillofacial radiology. Our goal is to evaluation the existing information just how the illness threatens patients and DHCPs and how to define which patients are possible to be SARS-CoV-2 infected; study how the usage of private shielding utensils and contamination control measures based on recent top observes, and knowledge can decrease the danger of virus spread in radiologic trials; and scrutinize how intraoral radiography, with its actually superior danger of scattering the infection, might be changed by extraoralradiographic methods for definite diagnostic jobs. In the pandemic, teleradiology has been extensively recycled for diagnostic determinations of COVID-19 patients, for discussions with radiologists in crisis cases, or managing of distance among radiology clinics. Dentists can have the digital radiographic images of their emergency patients through online service area also by electronic message or messaging applications to view in their smart phones, laptops, or other electronic devices.

Keywords: radiology, dental, oral, COVID-19, infection

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Authors: Helen Nelly Naiga, Jane Frances Zalwango, Saudah N. Kizito, Brian Agaba, Brenda N Simbwa, Maria Goretti Zalwango, Richard Migisha, Benon Kwesiga, Daniel Kadobera, Alex Ario Riolexus, Sarah Paige, Julie R. Harris

Abstract:

Background: Traditional community beliefs and practices can facilitate the spread of Ebola virus during outbreaks. On September 20, 2022, Uganda declared a Sudan Virus Disease (SVD) outbreak after a case was confirmed in Mubende District. During September–November 2022, the outbreak spread to eight additional districts. We investigated the role of community beliefs and practices in the spread of SUDV in Uganda in 2022. Methods: A qualitative study was conducted in Mubende, Kassanda, and Kyegegwa districts in February 2023. We conducted nine focus group discussions (FGDs) and six key informant interviews (KIIs). FGDs included SVD survivors, household members of SVD patients, traditional healers, religious leaders, and community leaders. Key informants included community, political, and religious leaders, traditional healers, and health workers. We asked about community beliefs and practices to understand if and how they contributed to the spread of SUDV. Interviews were recorded, translated, transcribed, and analyzed thematically. Results: Frequently-reported themes included beliefs that the community deaths, later found to be due to SVD, were the result of witchcraft or poisoning. Key informants reported that SVD patients frequently first consulted traditional healers or spiritual leaders before seeking formal healthcare, and noted that traditional healers treated patients with signs and symptoms of SVD without protective measures. Additional themes included religious leaders conducting laying-on-of-hands prayers for SVD patients and symptomatic contacts, SVD patients and their symptomatic contacts hiding in friends’ homes, and exhumation of SVD patients originally buried in safe and dignified burials, to enable traditional burials. Conclusion: Multiple community beliefs and practices likely promoted SVD outbreak spread during the 2022 outbreak in Uganda. Engaging traditional and spiritual healers early during similar outbreaks through risk communication and community engagement efforts could facilitate outbreak control. Targeted community messaging, including clear biological explanations for clusters of deaths and information on the dangers of exhuming bodies of SVD patients, could similarly facilitate improved control in future outbreaks in Uganda.

Keywords: Ebola, Sudan virus, outbreak, beliefs, traditional

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Authors: Rabah Laouar, Yahia Boudra, Adel Satouh, Adrian Boyce

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The ultramafic rocks of the Collo region crop out as « stratified » masses that cross-cut older metamorphic formation of the basement. These rocks are mainly peridotites and serpentinites. The peridotites are composed of olivine, orthopyroxene, clinopyroxene and spinel (chromite). The chemical composition of these lherzolites show a magnesian character with high MgO contents (34.4 to 37.5%), high Cr (0.14 to 0.27%), Ni (0.14 to 0.26%) and Co (34 to 133 ppm) and low CaO and Al₂O₃ (0.02 to 2.2 and 0.5 to 2.8 % respectively). They represent a residue (restite) of a mantle magmas partial melting. The chromite which represents about 2 to 3% of the rock is a ubiquitous mineral and shows two different generations: primary idiomorphic millimetric crystals and secondary very fine, xenomorphic and interstitial aggregates. The primary chromites are alumino-ferro-magnesian crystals. They show high Al₂O₃ (25.77% to 27.36%) and MgO (10.70% to 13.36%). Cr# (100*Cr/ (Al+Cr)) varies between 45 and 48, and Mg# (100*Mg/Mg+Fe₂+) varies between 49 and 59. On the other hand, the secondary interstitial grains are iron-rich chromites; they show low Al₂O₃ (4.67% to 9.54%) and MgO (4.60% to 4.65%). Cr# is relatively high (77 to 88) whereas Mg# show relatively low values, varying between 22 and 25. Oxygen isotopic composition of both types of chromites is consistent with their derivation from a mantle source (ð¹⁸O vary between +3.9 and +5.2‰), though a contribution of ¹⁶O-rich component to the secondary chromites is not ruled out.

Keywords: peridotites, serpentinites, chromite, partial melting, collo, Algeria

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Authors: Venuste Kayinamura, V. Iyamuremye, A. Ngirabakunzi

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Anti-retroviral therapy (ART) for HIV patient can cause a deficiency in glucose metabolism by promoting insulin resistance, glucose intolerance, and diabetes, diabetes mellitus keep increasing among HIV-infected patients worldwide but there is limited data on levels of blood glucose and its relationship with antiretroviral drugs (ARVs) and HIV-infection worldwide, particularly in Rwanda. A convenient sampling strategy was used in this study and it involved 323 HIV patients (n=323). Patients who are HIV positive under ARVs were involved in this study. The patient’s blood glucose was analyzed using an automated machine or glucometer (COBAS C 311). Data were analyzed using Microsoft Excel and SPSS V. 20.0 and presented in percentages. The highest diabetes mellitus prevalence was 93.33 % in people aged >40 years while the lowest diabetes mellitus prevalence was 6.67% in people aged between 21-and 40 years. The P-value was (0.021). Thus, there is a significant association between age and diabetes occurrence. The highest diabetes mellitus prevalence was 28.2% in patients under ART treatment for more than 10 years, 16.7% were <5years while 20% of patients were on ART treatment between 5-10 years. The P-value here is (0.03), thus the incidence of diabetes is associated with long-term ART use in HIV-infected patients. This study assessed the prevalence of diabetes among HIV-infected patients under ARVs attending the University Teaching Hospital of Butare (CHUB), it shows that the prevalence of diabetes is high in HIV-infected patients under ARTs. This study found no significant relationship between gender and diabetes mellitus growth. Therefore, regular assessment of diabetes mellitus especially among HIV-infected patients under ARVs is highly recommended to control other health issues caused by diabetes mellitus.

Keywords: anti-retroviral, diabetes mellitus, antiretroviral therapy, human immune deficiency virus

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Authors: Howaida I. Abd-Alla, Amal Z. Hassan, Maha Soltan, Atef G. Hanna, Mounir M. El-Safty

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Acokanthera oblongifolia (Apocynaceae) is used for treatment of several infection diseases and is a well-known cardiac glycoside-containing plant. The infusion of their leaves is gargled to treat tonsillitis and is used medicinally to treat snakebites. The total cardiac glycosides content in the leaves was determined by referring to gitoxigenin as a reference compound. Two triterpenes, lup-20(29)-en-3β-ol (1) and oleanolic acid (2); two cardenolides, acovenoside A (3) and acobioside A (4) were isolated from the ethyl acetate extract. Their structures were determined on the basis of spectral analysis. Major constituents isolated from this species were evaluated for cytotoxicity against normal lung cell line (Wi38) and antimicrobial activities against Gram-positive (two strains) and Gram-negative bacteria (four strains), yeast-like fungi (two strains) and fungi (five strains). The minimum inhibitory concentration (MIC) of the compounds was determined using broth microdilution method. Their viral inhibitory effects against avian influenza virus type A (AI-H5N1) and Newcastle disease virus (NDV) in specific pathogen free (SPF) embryonated chicken eggs (ECE), chicken embryo fibroblasts (CEF) and Vero cells were evaluated. The cardenolide (3) showed viral inhibitory effects against AI-H5N1 and NDV in SPF ECE. The two cardenolides isolated have shown potent cytotoxicity against Vero cells. Compound (3) showed potent anti-Gram-negative bacteria activity. These results suggested that acovenoside A might be promising for future antiviral and antimicrobial drug design.

Keywords: Acokanthera, AI-H5N1, Cardenolides, NDV, SPF-ECE, VERO, Wi38 , Microbe

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Authors: Gemma Postil, Claire Zanin, Michael Halpin, Caroline Ritter

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Illness experience research typically focuses on people that are living with a medical condition; however, the broad consequences of the COVID-19 pandemic are impacting those without the virus itself, as many experienced extensive lockdowns, social isolation, and distress. Drawing on conceptual work in the illness experience literature, we argue that policy and social changes tied to COVID-19 produce biographical disruptions. In this sense, we argue that the COVID-19 pandemic produces illness experience without illness, as the pandemic comprehensively impacts health and biography. This paper draws on 30 in-depth interviews with young adults living in Prince Edward Island (PEI), which were conducted as part of a larger project to understand how young adults navigate compliance with the COVID-19 pandemic. We then inductively analyzed the interviews with a constructivist grounded theory approach. Specifically, we demonstrate that young adults living in PEI during the COVID-19 pandemic experienced biographical disruptions throughout the pandemic despite not contracting the virus. First, we detail how some participants experience biographical acceleration, with the pandemic accelerating relationships, home buying, and career planning. Second, we demonstrate biographical stagnation, wherein participants report being unable to pursue major life milestones. Lastly, we describe biographical regression, wherein participants feel they are losing ground during the pandemic and are actively falling behind their peers. These findings provide the novel application of illness experience concepts to the context of the COVID-19 pandemic, contribute to work on illness experience and ambiguity, and extend Bury’s conceptualization of biographical disruption. In conclusion, we demonstrate that young adults experienced the biographical disruption expected from having COVID-19 without having an illness, highlighting the depth to which the pandemic affected young adults.

Keywords: illness experience, lived experience, biographical disruption, COVID-19, young adults

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Authors: Sara A. Al-Jaberi, Salma Ben-Salem, Meriam Messedi, Fatma Ayadi, Lihadh Al-Gazali, Bassam R. Ali

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Background: The chemokine receptor components play crucial roles in the immune system and some of them serve as co-receptors for the HIV virus. Several studies have documented those variants in chemokine receptors are correlated with susceptibility and resistance to infection with HIV virus. For example, mutations in the chemokine receptor 5 gene (CCR5) resulting in loss-of-function (such as the homozygous CCR5Δ32) confer high degree of resistance to HIV infection. Heterozygotes for these variants exhibit slow progression to AIDS. The prevalence of CCR5 polymorphisms varies among ethnic and geographical groups. For example, the CCR5 Δ32 variant is present in 10–15% of north Europeans but is rarely encountered among Africans. This study aims to identify the prevalence of some CCR5 variants in two geographically distant Arab populations (namely Emiratis and Tunisians). Methodology: The prevalence of CCR5 gene variants including CCR5Δ32, FS299, C101X, A29S and C178R has been determined using PCR and direct DNA sequencing. A total of 403 unrelated healthy individuals (253 Emiratis and 150 Tunisians) were genotyped for the CCR5Δ32 variant using PCR amplification and gel electrophoresis. In addition, 200 Emiratis have been screened for other SNPs using Sanger DNA sequencing. Results: Among Emiratis, the allele frequency of the CCR5Δ32 variant has been found to be 0.002. In addition, two variants L55Q and A159 were found at a frequency of 0.002.Moreover, the prevalence of the CCR5Δ32 variant in Tunisians was estimated to be 0.013 which is relatively higher than its frequency in Emiratis but lower than Europeans. Conclusion: We conclude that the allele frequency of the most critical CCR5 polymorphism (Δ32) is extremely low among Emiratis compared to other Arabs and North Europeans. In addition, very low allele frequencies of other CCR5 polymorphisms have been detected among Emiratis.

Keywords: chemokine receptors, CCR5Δ32, CCR5 polymorphisms, Emiratis, Arab populations

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Authors: Dong Tran, Thanh Dac Van, Ly Le

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Hemagglutinin (HA) and Neuraminidase (NA) play an important role in host immune evasion across influenza virus evolution process. The correlation between HA and NA evolution in respect to epitopic evolution and drug interaction has yet to be investigated. In this study, combining of sequence to structure evolution and statistical analysis on epitopic/binding site specificity, we identified potential therapeutic features of HA and NA that show specific antibody binding site of HA and specific binding distribution within NA active site of current inhibitors. Our approach introduces the use of sequence variation and molecular interaction to provide an effective strategy in establishing experimental based distributed representations of protein-protein/ligand complexes. The most important advantage of our method is that it does not require complete dataset of complexes but rather directly inferring feature interaction from sequence variation and molecular interaction. Using correlated sequence analysis, we additionally identified co-evolved mutations associated with maintaining HA/NA structural and functional variability toward immunity and therapeutic treatment. Our investigation on the HA binding specificity revealed unique conserved stalk domain interacts with unique loop domain of universal antibodies (CR9114, CT149, CR8043, CR8020, F16v3, CR6261, F10). On the other hand, NA inhibitors (Oseltamivir, Zaninamivir, Laninamivir) showed specific conserved residue contribution and similar to that of NA substrate (sialic acid) which can be exploited for drug design. Our study provides an important insight into rational design and identification of novel therapeutics targeting universally recognized feature of influenza HA/NA.

Keywords: influenza virus, hemagglutinin (HA), neuraminidase (NA), sequence evolution

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Authors: Thavasimuthu Citarasu, Mariavincent Michaelbabu, Vikram Vakharia

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The rhizome of Java grass, Cyperus rotundus was extracted different organic polar and non-polar solvents and performed the in vitro antiviral and immunostimulant activities against White Spot Syndrome Virus (WSSV) and Vibrio harveyi respectively. Based on the initial screening the ethyl acetate extract of C. rotundus was strong activities and further it was purified through silica column chromatography and the fractions were screened again for antiviral and immunostimulant activity. Among the different fractions screened against the WSSV and V. harveyi, the fractions, F-III to FV had strong activities. In order to study the in vivo influence of C. rotundus, the fractions (F-III to FV) were pooled and delivered to the F. indicus through artificial feed for 30 days. After the feeding trail the experimental and control diet fed F. indicus were challenged with virulent WSSV and studied the survival, molecular diagnosis, biochemical, haematological and immunological parameters. Surprisingly, the pooled fractions (F-III to FV) incorporated diets helped to significantly (P < 0.01) suppressed viral multiplication, showed significant (P < 0.01) differences in protein and glucose levels, improved total haemocyte count (THC), coagulase activity, significantly increased (P < =0.001) prophenol oxidase and intracellular superoxide anion production compared to the control shrimps. Based on the results, C. rotundus extracts effectively suppressed WSSV multiplication and improve the immune system in F. indicus against WSSV infection and this knowledge will helps to develop novel drugs from C. rotundus against WSSV.

Keywords: antiviral drugs, cyperus rotundus, fenneropenaeus indicus, WSSV

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Authors: Zainul A. Khan, Malik Z. Abdin, Jawaid A. Khan

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Begomoviruses belonging to the family Geminiviridae, have single-stranded circular DNA genomes that are monopartite or bipartite. The large intergenic region (LIR) of the monopartite and common region (CR) of bipartite begomoviruses possess promoter activity in their genomes. In this study, we have characterized novel bidirectional promoters from Cotton leaf curl Burewala virus (CLCuBuV) genome using high-throughput software and analyzed with PlantCARE, PLACE, Cister and PlantPAN databases. The promoters (Rep and CP promoters) were assayed both in stable and transient expression systems in tobacco as well as cotton plants. Rep and CP-based promoters from the LIR sequence of CLCuBuV and 35S promoter of Cauliflower mosaic virus (CaMV) were tagged with β-glucuronidase (GUS) and green fluorescent protein (GFP) reporter genes to check the efficacy of the promoters. Histochemical staining of GUS in transformed tobacco (Nicotiana tabacum cv. Xanthi) leaves showed higher GUS expression driven by CLCuBuV Rep (complimentary sense) promoter as compared to conventional CaMV 35S promoter and CLCuBuV CP (virion sense) promoter, respectively. GUS activity in individual plant cells driven by CLCuBuV Rep, CLCuBuV CP, and CaMV 35S promoter were quantified through fluorometric GUS assay and reverse transcription quantitative real-time PCR (RT-qPCR). The expression level of GUS tagged with CLCuBuV Rep promoter in the transformed tobacco plants was obtained 2 to 4 fold higher than CaMV 35S promoter. When CLCuBuV CP promoter was used, lower expression level was monitored than that by CaMV 35S promoter. The expression of GFP-tagged with CLCuBuV promoters was also investigated through agroinfiltration. The CLCuBuV Rep promoters showed stronger consistent transient expression in the leaves of N. benthamiana, N. tabacum and Gossypium hirsutum plants when compared with CaMV 35S and CLCuBuV CP promoter.

Keywords: Begmovirus, bidirectional promoter, CaMV 35S promoter, GFP, GUS, qPCR

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Authors: Masoud Soltanialvar, Ali Bagherpour

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Documented cases of human infection with H9N2 avian influenza viruses, first detected in 1999 in Hong Kong and China, indicate that these viruses can be directly transmitted from birds to humans. In this study, we characterized the mutation in the Hemagglutinin (HA) genes and proteins that correlates with a shift in affinity of the Hemagglutinin (HA) protein from the “avian” type sialic receptors to the “human” type in 10 Iranian isolates. We delineated the genomes and receptor binding profile of HA gene of some field isolates and established their phylogenetic relationship to the other Asian H9N2 sub lineages. A total of 1200 tissue samples collected from 40 farms located in various states of Iran during 2008 – 2010 as part of a program to monitor Avian Influenza Viruses (AIV) infection. To determine the genetic relationship of Iranian viruses, the Hemagglutinin (HA) genes from ten isolates were amplified and sequenced (by RT-PCR method). Nucleotide sequences (orf) of the (HA) genes were used for phylogenetic tree construction. Deduced amino acid sequences showed the presence of L226 (234 in H9 numbering) in all ten Iranian isolates which indicates a preference to binding of α (2–6) sialic acid receptors, so these Iranian H9N2 viruses have the potential to infect human beings. These isolates showed high degree of homology with 2 human H9N2 isolates A/HK/1073/99, A/HK/1074/99. Phylogenetic analysis of showed that all the HA genes of the Iranian H9N2 viruses fall into a single group within a G1-like sublineage which had contributed as donor of six internal genes to H5N1 highly pathogenic avian influenza. The results of this study indicated that all Iranian viruses have the potential to emerge as highly pathogenic influenza virus, and considering the homology of these isolates with human H9N2 strains, it seems that the potential of these avian influenza isolates to infect human should not be overlooked.

Keywords: influenza virus, hemagglutinin, neuraminidase, Iran

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Authors: Wajad Nazeer, Saghir Ahmad, Khalid Mahmood, Altaf Hussain, Abid Mahmood, Baoliang Zhou

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MNH-886(Bt.) is a upland cotton cultivar (Gossypium hirsutum L.) developed through hybridization of three parents [(FH-207×MNH-770)×Bollgard-1] at Cotton Research Station Multan, Pakistan. It is resistant to CLCuVD with 16.25 % disease incidence (60 DAS, March sowing) whereas moderately susceptible to CLCuVD when planted in June with disease incidence 34 % (60 DAS). This disease reaction was lowest among 25 cotton advanced lines/varieties tested at hot spots of CLCuVD. Its performance was tested during 2009 to 2012 in various indigenous, provincial, and national varietal trials in comparison with the commercial variety IR-3701 and AA-802 & CIM-496. In PCCT trial during 2009-10; 2011-12, MNH-886 surpassed all the existing Bt. strains along with commercial varieties across the Punjab province with seed cotton yield production 2658 kg ha-1 and 2848 kg ha-1 which was 81.31 and 13% higher than checks, respectively. In National Coordinated Bt. Trial, MNH-886(Bt.) produced 3347 kg ha-1 seed cotton at CCRI, Multan; the hot spot of CLCuVD, in comparison to IR-3701 which gave 2556 kg ha-1. It possesses higher lint percentage (41.01%), along with the most desirable fibre traits (staple length 28.210mm, micronaire value 4.95 µg inch-1 and fibre strength 99.5 tppsi, and uniformity ratio 82.0%). The quantification of toxicity level of crystal protein was found positive for Cry1Ab/Ac protein with toxicity level 2.76µg g-1 and Mon 531 event was confirmed. Having tremendous yield potential, good fibre traits, and great tolerance to CLCuVD we can recommended this variety for cultivation in CLCuVD hotspots of Pakistan.

Keywords: cotton, cultivar, cotton leaf curl virus, CLCuVD hit districts

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Authors: Abbas R. Ali, Daroon Hasan Khorsheed

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Twenty one samples of stream sediments were collected from the Diyala River Basin (DRB), which represent one of three major tributaries of the Tigris River at northeastern Iraq. This study is concerned with the heavy minerals (HM) analysis in the + 63μ m fraction of the Diyala River sediments, distribution pattern in the various river basin sectors, as well as comparing the present results with previous works.The metastable heavy minerals (epidote, staurolite, garnet) represent more than (30%) Whereas the ultrastable heavy minerals (pyroxene and amphibole) make only about (19 %). Opaques are present in high proportions reaching about (29%) as an average. The ultrastable (zircon, tourmaline, rutile) heavy minerals are the miner constituents (7%) in the sediments.According to the laboratory analytical data of heavy mineral distributions the studied sediments are derived from mafic and ultramafic rocks are found in northeastern Iraq that represent Walash – Nawpordan Series and Mawat complexes in Zagros zones. The presence of zircon and tourmaline in trace amounts may give an indication for the weak role of acidic rocks in the source area whereas the epidote group minerals give an indication for the role of metamorphic rocks.

Keywords: heavy minerals, mineral distribution, recent stream sediment, Diyala river, northeastern Iraq

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Authors: David Shengelia, Giorgi Chichinadze, Tamara Tsutsunava, Giorgi Beridze, Irakli Javakhishvili

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The Loki pre-Alpine crystalline massif crops out within the Caucasus region. The massif basement is represented by the Upper Devonian gneissose quartz-diorites, the Lower-Middle Paleozoic metamorphic allochthonous complex, and different magmatites. Earlier, the metamorphic complex was considered as indivisible set represented by the series of different temperature metamorphits. The degree of metamorphism of separate parts of the complex is due to different formation conditions. This fact according to authors of the abstract was explained by the allochthonous-flaky structure of the complex. It was stated that the complex thrust over the gneissose quartz diorites before the intrusion of Sudetic granites. During the detailed mapping, the authors turned out that the metamorphism issues need to be reviewed and additional researches to be carried out. Investigations were accomplished by using the following methodologies: finding of key sections, a sampling of rocks, microscopic description of the material, analytical determination of elements in the rocks, microprobe analysis of minerals and new interpretation of obtained data. According to the author’s recent data within the massif four tectonic plates: Lower Gorastskali, Sapharlo-Lok-Jandari, Moshevani and “mélange” overthrust sheets have been mapped. They differ from each other by composition, the degree of metamorphism and internal structure. It is confirmed that the initial rocks of the tectonic plates formed in different geodynamic conditions during overthrusting due to tectonic compression form a thick tectonic sheet. Based on the detailed laboratory investigations additional mineral assemblages were established, temperature limits were specified, and a renewed trend of metamorphism facies and subfacies was elaborated. The results are the following: 1. The Lower Gorastskali overthrust sheet is a fragment of ophiolitic association corresponding to the Paleotethys oceanic crust. The main rock-forming minerals are carbonate, chlorite, spinel, epidote, clinoptilolite, plagioclase, hornblende, actinolite, hornblende, albite, serpentine, tremolite, talc, garnet, and prehnite. Regional metamorphism of rocks corresponds to the greenschist facies lowest stage. 2. The Sapharlo-Lok-Jandari overthrust sheet metapelites are represented by chloritoid, chlorite, phengite, muscovite, biotite, garnet, ankerite, carbonate, and quartz. Metabasites containing actinolite, chlorite, plagioclase, calcite, epidote, albite, actinolitic hornblende and hornblende are also present. The degree of metamorphism corresponds to the greenschist high-temperature chlorite, biotite, and low-temperature garnet subfacies. Later the rocks underwent the contact influence of Late Variscan granites. 3. The Moshevani overthrust sheet is represented mainly by metapelites and rarely by metabasites. Main rock-forming minerals of metapelites are muscovite, biotite, chlorite, quartz, andalusite, plagioclase, garnet and cordierite and of metabasites - plagioclase, green and blue-green hornblende, chlorite, epidote, actinolite, albite, and carbonate. Metamorphism level corresponds to staurolite-andalusite subfacies of staurolite facies and partially to facies of biotite muscovite gneisses and hornfelse facies as well. 4. The “mélange” overthrust sheet is built of different size rock fragments and blocks of Moshevani and Lower Gorastskali overthrust sheets. The degree of regional metamorphism of first and second overthrust sheets of the Loki massif corresponds to chlorite, biotite, and low-temperature garnet subfacies, but of the third overthrust sheet – to staurolite-andalusite subfacies of staurolite facies and partially to facies of biotite muscovite gneisses and hornfelse facies.

Keywords: regional metamorphism, crystalline massif, mineral assemblages, the Caucasus

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Authors: Alessandro Poma, Kal Karim, Sergey Piletsky, Giuseppe Battaglia

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The recent increasing emergency threat to public health of infectious influenza diseases has prompted interest in the detection of avian influenza virus (AIV) H5N1 in humans as well as animals. A variety of technologies for diagnosing AIV infection have been developed. However, various disadvantages (costs, lengthy analyses, and need for high-containment facilities) make these methods less than ideal in their practical application. Molecularly Imprinted Polymeric Nanoparticles (MIP NPs) are suitable to overcome these limitations by having high affinity, selectivity, versatility, scalability and cost-effectiveness with the versatility of post-modification (labeling – fluorescent, magnetic, optical) opening the way to the potential introduction of improved diagnostic tests capable of providing rapid differential diagnosis. Here we present our first results in the production and testing of MIP NPs for the detection of AIV H5N1. Recent developments in the solid-phase synthesis of MIP NPs mean that for the first time a reliable supply of ‘soluble’ synthetic antibodies can be made available for testing as potential biological or diagnostic active molecules. The MIP NPs have the potential to detect viruses that are widely circulating in farm animals and indeed humans. Early and accurate identification of the infectious agent will expedite appropriate control measures. Thus, diagnosis at an early stage of infection of a herd or flock or individual maximizes the efficiency with which containment, prevention and possibly treatment strategies can be implemented. More importantly, substantiating the practicability’s of these novel reagents should lead to an initial reduction and eventually to a potential total replacement of animals, both large and small, to raise such specific serological materials.

Keywords: influenza virus, molecular imprinting, nanoparticles, polymers

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Authors: Olga V. Chervyakova, Elmira T. Tailakova, Vitaliy M. Strochkov, Kulyaisan T. Sultankulova, Nurlan T. Sandybayev, Lev G. Nemchinov, Rosemarie W. Hammond

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Sheep pox is a highly contagious infection that OIE regards to be one of the most dangerous animal diseases. It causes enormous economic losses because of death and slaughter of infected animals, lower productivity, cost of veterinary and sanitary as well as quarantine measures. To control spread of sheep pox infection the attenuated vaccines are widely used in the Republic of Kazakhstan and other Former Soviet Union countries. In spite of high efficiency of live vaccines, the possible presence of the residual virulence, potential genetic instability restricts their use in disease-free areas that leads to necessity to exploit new approaches in vaccine development involving recombinant DNA technology. Vaccines on the basis of recombinant proteins are the newest generation of prophylactic preparations. The main advantage of these vaccines is their low reactogenicity and this fact makes them widely used in medical and veterinary practice for vaccination of humans and farm animals. The objective of the study is to produce recombinant immunogenic proteins for development of the high-performance means for sheep pox prophylaxis. The SPV proteins were chosen for their homology with the known immunogenic vaccinia virus proteins. Assay of nucleotide and amino acid sequences of the target SPV protein genes. It has been shown that four proteins SPPV060 (ortholog L1), SPPV074 (ortholog H3), SPPV122 (ortholog A33) and SPPV141 (ortholog B5) possess transmembrane domains at N- or C-terminus while in amino acid sequences of SPPV095 (ortholog А 4) and SPPV117 (ortholog А 27) proteins these domains were absent. On the basis of these findings the primers were constructed. Target genes were amplified and subsequently cloned into the expression vector рЕТ26b(+) or рЕТ28b(+). Six constructions (pSPPV060ΔТМ, pSPPV074ΔТМ, pSPPV095, pSPPV117, pSPPV122ΔТМ and pSPPV141ΔТМ) were obtained for expression of the SPV genes under control of T7 promoter in Escherichia coli. To purify and detect recombinant proteins the amino acid sequences were modified by adding six histidine molecules at C-terminus. Induction of gene expression by IPTG was resulted in production of the proteins with molecular weights corresponding to the estimated values for SPPV060, SPPV074, SPPV095, SPPV117, SPPV122 and SPPV141, i.e. 22, 30, 20, 19, 17 and 22 kDa respectively. Optimal protocol of expression for each gene that ensures high yield of the recombinant protein was identified. Assay of cellular lysates by western blotting confirmed expression of the target proteins. Recombinant proteins bind specifically with antibodies to polyhistidine. Moreover all produced proteins are specifically recognized by the serum from experimentally SPV-infected sheep. The recombinant proteins SPPV060, SPPV074, SPPV117, SPPV122 and SPPV141 were also shown to induce formation of antibodies with virus-neutralizing activity. The results of the research will help to develop a new-generation high-performance means for specific sheep pox prophylaxis that is one of key moments in animal health protection. The research was conducted under the International project ISTC # K-1704 “Development of methods to construct recombinant prophylactic means for sheep pox with use of transgenic plants” and under the Grant Project RK MES G.2015/0115RK01983 "Recombinant vaccine for sheep pox prophylaxis".

Keywords: prophylactic preparation, recombinant protein, sheep pox virus, subunit vaccine

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Authors: Aussara Panya, Nunghathai Sawasdee, Mutita Junking, Chatchawan Srisawat, Kiattawee Choowongkomon, Pa-Thai Yenchitsomanus

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Dengue virus (DENV) infection is a global public health problem with approximately 100 million infected cases a year. Presently, there is no approved vaccine or effective drug available; therefore, the development of anti-DENV drug is urgently needed. The clinical reports revealing the positive association between the disease severity and viral titer has been reported previously suggesting that the anti-DENV drug therapy can possibly ameliorate the disease severity. Although several anti-DENV agents showed inhibitory activities against DENV infection, to date none of them accomplishes clinical use in the patients. The surface envelope (E) protein of DENV is critical for the viral entry step, which includes attachment and membrane fusion; thus, the blocking of envelope protein is an attractive strategy for anti-DENV drug development. To search the safe anti-DENV agent, this study aimed to search for novel peptide inhibitors to counter DENV infection through the targeting of E protein using a structure-based in silico design. Two selected strategies has been used including to identify the peptide inhibitor which interfere the membrane fusion process whereby the hydrophobic pocket on the E protein was the target, the destabilization of virion structure organization through the disruption of the interaction between the envelope and membrane proteins, respectively. The molecular docking technique has been used in the first strategy to search for the peptide inhibitors that specifically bind to the hydrophobic pocket. The second strategy, the peptide inhibitor has been designed to mimic the ectodomain portion of membrane protein to disrupt the protein-protein interaction. The designed peptides were tested for the effects on cell viability to measure the toxic to peptide to the cells and their inhibitory assay to inhibit the DENV infection in Vero cells. Furthermore, their antiviral effects on viral replication, intracellular protein level and viral production have been observed by using the qPCR, cell-based flavivirus immunodetection and immunofluorescence assay. None of tested peptides showed the significant effect on cell viability. The small peptide inhibitors achieved from molecular docking, Glu-Phe (EF), effectively inhibited DENV infection in cell culture system. Its most potential effect was observed for DENV2 with a half maximal inhibition concentration (IC50) of 96 μM, but it partially inhibited other serotypes. Treatment of EF at 200 µM on infected cells also significantly reduced the viral genome and protein to 83.47% and 84.15%, respectively, corresponding to the reduction of infected cell numbers. An additional approach was carried out by using peptide mimicking membrane (M) protein, namely MLH40. Treatment of MLH40 caused the reduction of foci formation in four individual DENV serotype (DENV1-4) with IC50 of 24-31 μM. Further characterization suggested that the MLH40 specifically blocked viral attachment to host membrane, and treatment with 100 μM could diminish 80% of viral attachment. In summary, targeting the hydrophobic pocket and M-binding site on the E protein by using the peptide inhibitors could inhibit DENV infection. The results provide proof of-concept for the development of antiviral therapeutic peptide inhibitors to counter DENV infection through the use of a structure-based design targeting conserved viral protein.

Keywords: dengue virus, dengue virus infection, drug design, peptide inhibitor

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Authors: Aniekan Peter, ECS Naidu, Edidiong Akang, U. Offor, R. Kalhapure, A. A. Chuturgoon, T. Govender, O. O. Azu

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Introduction: Nano-drugs are novel innovations in the management of human immunodeficiency virus (HIV) pandemic, especially resistant strains of the virus in their sanctuary sites: testis and the brain. There are safety concerns to be addressed to achieve the full potential of this new drug delivery system. Aim of study: Our study was designed to investigate toxicity profile of Tenofovir Nanoparticle (TDF-N) synthesized by University of Kwazulu-Natal (UKZN) Nano-team for prevention and treatment of HIV infection. Methodology: Ten adult male Sprague-Dawley rats maintained at the Animal House of the Biomedical Resources Unit UKZN were used for the study. The animals were weighed and divided into two groups of 5 animal each. Control animals (A) were administered with normal saline. Therapeutic dose (4.3 mg/kg) of TDF-N was administered to group B. At the end of four weeks, animals were weighed and sacrificed. Liver and kidney were removed fixed in formal saline, processed and stained using H/E, PAS and MT stains for light microscopy. Serum was obtained for renal function test (RFT), liver function test (LFT) and full blood count (FBC) using appropriate analysers. Cellular measurements were done using ImageJ and Leica software 2.0. Data were analysed using graph pad 6, values < 0.05 were significant. Results: We reported no histological alterations in the liver, kidney, FBC, LFT and RFT between the TDF-N animals and saline control. There were no significant differences in weight, organo-somatic index and histological measurements in the treatment group when compared with saline control. Conclusion/recommendations: TDF-N is not toxic to the liver, kidney and blood cells in our study. More studies using human subjects is recommended.

Keywords: tenofovir nanoparticles, liver, kidney, blood cells

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Authors: Ibrahim Alreshidi, Aseel Albrekeit, Ruaa Alharthi

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Background: As a result of the spread of COVID-19 at the beginning of 2020, many governments, including Saudi Arabia, have suspended operations in many agencies. Most dermatologists have restricted their practice, including cosmetic procedures, to ensure social distancing. On the 7th of May 2020, Saudi authorities reduced the restriction of COVID-19 virus preventative measures, allowing clinics to start accepting patients following the ministry of health protocols. Objectives: Evaluation of the public's perception and willingness to undergo cosmetic procedures during COVID-19 outbreaks in Saudi Arabia. Materials and methods: A descriptive, cross-sectional, questionnaire-based study was carried out among the individuals who lack typical symptoms of COVID-19 infection in Saudi Arabia. A self-designed web-based questionnaire was developed; content face validity and a pilot study were done. The questionnaire was distributed electronically from the 8th of May until the 31st of May 2020. Results: A total of 656 individuals who lack typical symptoms of COVID-19 infection were included in this analysis. Only 10.5% of participants expressed their will to do cosmetic procedures during the COVID-19 pandemic. More than 90% of the participants believed that the COVID-19 pandemic was either somewhat serious (52.9%) or very serious (38.7%). The willingness to do cosmetic procedures during the COVID-19 pandemic remained unaltered when the price was discounted (p<0.001) and when infection control measures were ensured (p<0.001). Conclusion: The COVID-19 pandemic had a negative impact on the practice of cosmetic dermatology. Fear of transferring the infection to a beloved home member is the main reason to avoid these procedures. Generating well-structured safety guidelines to decrease the risk of this unusual virus transmission in dermatology practice and creating financial incentives may help increase the public willingness to do these cosmetic procedures during this pandemic.

Keywords: COVID-19 pandemic, cosmetic procedures, questionnaire, dermatology

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Authors: Ali Kassem

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Background: In the last few years, factors such as insulin resistance (IR) and hepatic steatosis have been linked to progression of hepatic fibrosis.Patients with chronic liver disease, and cirrhosis in particular, are known to be prone to IR. However, chronic HCV (hepatitis C) infection may induce IR, regardless of the presence of liver cirrhosis. Our aims are to study insulin resistance (IR) assessed by HOMA-IR (Homeostatic Model Assessment Insulin Resistance) as a possible risk factor in disease progression in cirrhotic patients and to evaluate the role of IR in hepatic fibrosis progression. The correlations of HOMA-IR values to laboratory, virological and histopathological parameters of chronic HCV are also examined. Methods: The study included 50 people divided into 30 adult chronic hepatitis C patients diagnosed by PCR (polymerase chain reaction) within previous 6 months and 20 healthy controls. The functional and morphological status of the liver were evaluated by ultrasonography and laboratory investigations including liver function tests and by liver biopsy. Fasting blood glucose and fasting insulin levels were measured and body mass index and insulin resistance were calculated. Patients having HOMA-IR >2.5 were labeled as insulin resistant. Results: Chronic hepatitis C patients with IR showed significantly higher mean values of BMI (body mass index) and fasting insulin than those without IR (P < 0.000). Patients with IR were more likely to have steatosis (p = 0.006), higher necroinflammatory activity (p = 0.05). No significant differences were found between the two groups regarding hepatic fibrosis. Conclusion: HOMA-IR measurement could represent a novel marker to identify the cirrhotic patients at greater risk for the progression of liver disease. As IR is a potentially modifiable risk factor, these findings may have important prognostic and therapeutic implications. Assessment of IR by HOMA-IR and improving insulin sensitivity are recommended in patients with HCV and related chronic liver disease.

Keywords: hepatic fibrosis, hepatitis C virus infection, hepatic steatosis, insulin resistance

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Authors: Wafa Toumi, Alessandro Ripalti, Luigi Ricciardiello, Dalila Gargouri, Jamel Kharrat, Abderraouf Cherif, Ahmed Bouhafa, Slim Jarboui, Mohamed Zili, Ridha Khelifa

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Background & aims: Colorectal cancer (CRC) is one of the most common malignancies throughout the world. Several risk factors, both genetic and environmental, including viral infections, have been linked to colorectal carcinogenesis. A few studies report the detection of human polyomavirus JC (JCV) DNA and transformation antigen (T-Ag) in a fraction of the colorectal tumors studied and suggest an association of this virus with CRC. In order to investigate whether such an association of JCV with CRC will hold in a different epidemiological setting, we looked for the presence of JCV DNA and T-Ag expression in a group of Tunisian CRC patients. Methods: Fresh colorectal mucosa biopsies were obtained from 17 healthy volunteers and from both colorectal tumors and adjacent normal tissues of 47 CRC patients. DNA was extracted from fresh biopsies or from formalin-fixed, paraffin-embedded tissue sections using the Invitrogen Purelink Genomic DNA mini Kit. A simple PCR and a nested PCR were used to amplify a region of the T-Ag gene. The obtained PCR products revealed a 154 bp and a 98 bp bands, respectively. Specificity was confirmed by sequencing of the PCR products. T-Ag expression was determined by immunohistochemical staining using a mouse monoclonal antibody (clone PAb416) directed against SV40 T-Ag that cross reacts with JCV T-Ag. Results: JCV DNA was found in 12 (25%) and 22 (46%) of the CRC tumors by simple PCR and by nested PCR, respectively. All paired adjacent normal mucosa biopsies were negative for viral DNA. Sequencing of the DNA amplicons obtained confirmed the authenticity of T-Ag sequences. Immunohistochemical staining showed nuclear T-Ag expression in all 22 JCV DNA- positive samples and in 3 additional tumor samples which appeared DNA-negative by PCR. Conclusions: These results suggest an association of JCV with a subpopulation of Tunisian colorectal tumors.

Keywords: colorectal cancer, immunohistochemistry, Polyomavirus JC, PCR

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

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

Keywords: dengue, lncRNA, NEAT1, transcriptome

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Authors: Pradakshina Sharma, Jagriti Narang

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Due to the critical significance in the early identification of infectious diseases, electrochemical sensors have garnered considerable interest. Here, we develop a detection platform for the chikungunya virus by rationally implementing the extremely high charge-transfer efficiency of a ternary nanocomposite of graphene oxide, silver, and gold (G/Ag/Au) (CHIKV). Because paper is an inexpensive substrate and can be produced in large quantities, the use of electrochemical paper analytical device (EPAD) origami further enhances the sensor's appealing qualities. A cost-effective platform for point-of-care diagnostics is provided by paper-based testing. These types of sensors are referred to as eco-designed analytical tools due to their efficient production, usage of the eco-friendly substrate, and potential to reduce waste management after measuring by incinerating the sensor. In this research, the paper's foldability property has been used to develop and create 3D multifaceted biosensors that can specifically detect the CHIKVX-ray diffraction, scanning electron microscopy, UV-vis spectroscopy, and transmission electron microscopy (TEM) were used to characterize the produced nanoparticles. In this work, aptamers are used since they are thought to be a unique and sensitive tool for use in rapid diagnostic methods. Cyclic voltammetry (CV) and linear sweep voltammetry (LSV), which were both validated with a potentiostat, were used to measure the analytical response of the biosensor. The target CHIKV antigen was hybridized with using the aptamer-modified electrode as a signal modulation platform, and its presence was determined by a decline in the current produced by its interaction with an anionic mediator, Methylene Blue (MB). Additionally, a detection limit of 1ng/ml and a broad linear range of 1ng/ml-10µg/ml for the CHIKV antigen were reported.

Keywords: biosensors, ePAD, arboviral infections, point of care

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Authors: J. T. Koininga, J. E. Teigen, A. Wilkinson, D. Kanneh, F. Kanneh, M. Foday, D. S. Grant, M. Leach, L. M. Moses

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Lassa fever (LF) is a severe febrile illness endemic to West Africa. While human-to-human transmission occurs, evidence suggests most LF cases originate from exposure to rodents, particularly the Natal multimammate mouse, Mastomys natalensis. Within West Africa, LF occurs primarily in rural communities where agriculture is the main economic activity. Seasonality of LF has also been linked to agricultural cycles, with peak incidence occurring in the dry season when fields are burned and plowed. To investigate this pattern of seasonality, four agricultural communities were selected for this two-year longitudinal study. Each community was to be sampled four times each year, but this was interrupted by the Ebola virus disease outbreak. Agricultural land use, forested, and fallow areas were identified through participatory mapping. Transects were plotted in each area and Sherman traps were set for four nights. Captured small mammals were identified, ear tagged, and released. Mastomys natalensis abundance was found to be highest in areas of converted fallow land and rice swamps in the dry season and upland mixed crop areas toward the onset of the rainy season. All peak times were associated with heavy perturbation of soil. All ages and genders were present during these time points. These results suggest that peak abundance of the Mastomys natalensis in agricultural areas coincides with peak incidence of LF reported in this region. Although contact with rodents may be higher in villages, our study suggests human behaviors in agricultural areas may increase risk of transmission of Lassa virus.

Keywords: agriculture, land use, Lassa Fever, rodent abundance

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