Search results for: bovine papilloma virus
508 Evaluation of Medicinal Plants, Catunaregam spinosa, Houttuynia cordata, and Rhapis excelsa from Malaysia for Antibacterial, Antifungal and Antiviral Properties
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
Procedia PDF Downloads 403507 Association of Temperature Factors with Seropositive Results against Selected Pathogens in Dairy Cow Herds from Central and Northern Greece
Authors: Marina Sofia, Alexios Giannakopoulos, Antonia Touloudi, Dimitris C Chatzopoulos, Zoi Athanasakopoulou, Vassiliki Spyrou, Charalambos Billinis
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Fertility of dairy cattle can be affected by heat stress when the ambient temperature increases above 30°C and the relative humidity ranges from 35% to 50%. The present study was conducted on dairy cattle farms during summer months in Greece and aimed to identify the serological profile against pathogens that could affect fertility and to associate the positive serological results at herd level with temperature factors. A total of 323 serum samples were collected from clinically healthy dairy cows of 8 herds, located in Central and Northern Greece. ELISA tests were performed to detect antibodies against selected pathogens that affect fertility, namely Chlamydophila abortus, Coxiella burnetii, Neospora caninum, Toxoplasma gondii and Infectious Bovine Rhinotracheitis Virus (IBRV). Eleven climatic variables were derived from the WorldClim version 1.4. and ArcGIS V.10.1 software was used for analysis of the spatial information. Five different MaxEnt models were applied to associate the temperature variables with the locations of seropositive Chl. abortus, C. burnetii, N. caninum, T. gondii and IBRV herds (one for each pathogen). The logistic outputs were used for the interpretation of the results. ROC analyses were performed to evaluate the goodness of fit of the models’ predictions. Jackknife tests were used to identify the variables with a substantial contribution to each model. The seropositivity rates of pathogens varied among the 8 herds (0.85-4.76% for Chl. abortus, 4.76-62.71% for N. caninum, 3.8-43.47% for C. burnetii, 4.76-39.28% for T. gondii and 47.83-78.57% for IBRV). The variables of annual temperature range, mean diurnal range and maximum temperature of the warmest month gave a contribution to all five models. The regularized training gains, the training AUCs and the unregularized training gains were estimated. The mean diurnal range gave the highest gain when used in isolation and decreased the gain the most when it was omitted in the two models for seropositive Chl.abortus and IBRV herds. The annual temperature range increased the gain when used alone and decreased the gain the most when it was omitted in the models for seropositive C. burnetii, N. caninum and T. gondii herds. In conclusion, antibodies against Chl. abortus, C. burnetii, N. caninum, T. gondii and IBRV were detected in most herds suggesting circulation of pathogens that could cause infertility. The results of the spatial analyses demonstrated that the annual temperature range, mean diurnal range and maximum temperature of the warmest month could affect positively the possible pathogens’ presence. Acknowledgment: This research has been co‐financed by the European Regional Development Fund of the European Union and Greek national funds through the Operational Program Competitiveness, Entrepreneurship and Innovation, under the call RESEARCH–CREATE–INNOVATE (project code: T1EDK-01078).Keywords: dairy cows, seropositivity, spatial analysis, temperature factors
Procedia PDF Downloads 198506 Screening of Different Native Genotypes of Broadleaf Mustard against Different Diseases
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
Procedia PDF Downloads 80505 Integration of FMEA and Human Factor in the Food Chain Risk Assessment
Authors: Mohsen Shirani, Micaela Demichela
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During the last decades, a number of food crises such as Bovine Spongiform Encephalopathy (BSE), Mad-Cow disease, Dioxin in chicken food, Food-and-Mouth Disease (FMD), have certainly inflicted the reliability of the food industry. Consequently, the trend in applying different scientific methods of risk assessment in food safety has obtained more attentions in the academic and practice. However, lack of practical approach considering entire food supply chain is tangible in the academic literature. In this regard, this paper aims to apply risk assessment tool (FMEA) with integration of Human Factor along the entire supply chain of food production and test the method in a case study of Diary production, and analyze its results.Keywords: FMEA, food supply chain, risk assessment, human factor
Procedia PDF Downloads 447504 Development of an Aerosol Protection Capsule for Patients with COVID-19
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
Procedia PDF Downloads 84503 Oncolytic H-1 Parvovirus Entry in Cancer Cells through Clathrin-Mediated Endocytosis
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
Procedia PDF Downloads 155502 Role of Tyrosine-Phosphorylated STAT3 in Liver Regeneration: Survival, DNA Synthesis, Inflammatory Reaction and Liver Mass Recovery
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
Procedia PDF Downloads 312501 Comprehensive Evaluation of Oral and Maxillofacial Radiology in "COVID-19"
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
Procedia PDF Downloads 172500 The Role of Community Beliefs and Practices on the Spread of Ebola in Uganda, September 2022
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
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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
Procedia PDF Downloads 55499 Prevalence of Bovine Mastitis and Associated Risk Factors in Selected Dairy Farms in Zoba Anseba, Eritrea
Authors: Redie Kidane Ghebrehawariat, Betiel Habte Hadgu, Filmon Berhane Kahsay, Rim Berhane Fisehaye, Samuel Haile Kahsay, Saron Yemane Yosief, Selemawit Mosazghi Gilazghi
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A cross-sectional study was conducted from 22 February to 9 April 2022 on small, medium, and large holder dairy farms to determine the bovine mastitis prevalence and associated risk factors in the Anseba region, Eritrea. A total of 34 dairy farms and 193 dairy cows were randomly selected. Dairy cows were physically examined for any change on the udder and milk; a California mastitis test was performed to check sub-clinical mastitis; a closed-ended semi-structured questionnaire composed of 28 variables/risk factors (21 management risk factors and 7 animal-level risk factors) was used to determine the risk factors responsible for clinical and sub-clinical mastitis in the dairy cows. The overall cow-level prevalence of mastitis was 147 (76.2%). The animal level prevalence rate of clinical and sub-clinical mastitis was found to be 22 (11.4%) and 125 (64.8%), respectively, while herd level prevalence both for clinical and subclinical mastitis was found to be 14 (41.2%) and 26 (76.5%) respectively. Based on the already set P-value, which is <0.05, a number of risk factors were found to have a significant relationship with the occurrence of clinical and sub-clinical mastitis. Generally, animal risk factors such as animal age, parity, injury on the udder or teat, and previous history of mastitis presence of injury on the udder and lactation stage were risk factors with a significant relationship with the occurrence of clinical and sub-clinical mastitis. On the other hand, management risk factors with a significant relationship to the occurrence of clinical and sub-clinical mastitis were herd size, failure to milk mastitic cow, at last, educational level, floor type, failure to use a towel, using one towel for more than one cow and failure to practice mastitis test. From a total of 772 quarters, 280 (36.3%) were found positive for sub-clinical mastitis using the California mastitis test; of these, 70 (9%) were weakly positive, 90 (11.7%) were distinct positive, and 120 (15.5%) were strongly positive. Furthermore, 13 (1.7%) quarters were blocked. Quarter level prevalence was right front 80 (41.5%), left front 64 (33.3%), right hind 69 (35.8%) and left hind 67 (34.7%). The study has shown that mastitis is a major problem for dairy farms and the findings suggested that mastitis is one of the limiting factors in increasing milk production. Subclinical mastitis was found to be a devastating problem, and it occurred in all three breeds of lactating dairy cattle. Therefore, farmers should work hard to avoid the above-mentioned risk factors to minimize the infection of their dairy cattle by mastitis and thereby increase their profit. On the other hand, the Ministry of Agriculture, through the extension unit, should work in close contact with the farmers to increase awareness of the economic importance of the disease and associated risk factors.Keywords: mastitis, prevalence, dairy cattle, Anseba, Eritrea
Procedia PDF Downloads 128498 Prevalence of Diabetes Mellitus Among Human Immune Deficiency Virus-Positive Patients Under Anti-retroviral Attending in Rwanda, a Case Study of University Teaching Hospital of Butare
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
Procedia PDF Downloads 113497 Cytotoxic, Antimicrobial and Antiviral Activities of Acovenoside A: A Cardenolide Isolated from an Egyptian Cultivar of Acokanthera spectabilis Leaves
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
Procedia PDF Downloads 178496 Illness Experience Without Illness: A Qualitative Study on the Lived Experience of Young Adults During the COVID-19 Pandemic
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
Procedia PDF Downloads 161495 Determination of the CCR5Δ32 Frequency in Emiratis and Tunisians and Screening of the CCR5 Gene for Novel Alleles in Emiratis
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
Procedia PDF Downloads 378494 In-Depth Analysis on Sequence Evolution and Molecular Interaction of Influenza Receptors (Hemagglutinin and Neuraminidase)
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
Procedia PDF Downloads 164493 Influence of Cyperus Rotundus Active Principles Inhibit Viral Multiplication and Stimulate Immune System in Indian White Shrimp Fenneropenaeus Indicus against White Spot Syndrome Virus Infection
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
Procedia PDF Downloads 456492 Comparison of Two Methods of Cryopreservation of Testicular Tissue from Prepubertal Lambs
Authors: Rensson Homero Celiz Ygnacio, Marco Aurélio Schiavo Novaes, Lucy Vanessa Sulca Ñaupas, Ana Paula Ribeiro Rodrigues
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The cryopreservation of testicular tissue emerges as an alternative for the preservation of the reproductive potential of individuals who still cannot produce sperm; however, they will undergo treatments that may affect their fertility (e.g., chemotherapy). Therefore, the present work aims to compare two cryopreservation methods (slow freezing and vitrification) in testicular tissue of prepubertal lambs. For that, to obtain the testicular tissue, the animals were castrated and the testicles were collected immediately in a physiological solution supplemented with antibiotics. In the laboratory, the testis was split into small pieces. The total size of the testicular fragments was 3×3x1 mm³ and was placed in a dish contained in Minimum Essential Medium (MEM-HEPES). The fragments were distributed randomly into non-cryopreserved (fresh control), slow freezing (SF), and vitrified. To SF procedures, two fragments from a given male were then placed in a 2,0 mL cryogenic vial containing 1,0 mL MEM-HEPES supplemented with 20% fetal bovine serum (FBS) and 20% dimethylsulfoxide (DMSO). Tubes were placed into a Mr. Frosty™ Freezing container with isopropyl alcohol and transferred to a -80°C freezer for overnight storage. On the next day, each tube was plunged into liquid nitrogen (NL). For vitrification, the ovarian tissue cryosystem (OTC) device was used. Testicular fragments were placed in the OTC device and exposed to the first vitrification solution composed of MEM-HEPES supplemented with 10 mg/mL Bovine Serum Albumin (BSA), 0.25 M sucrose, 10% Ethylene glycol (EG), 10% DMSO and 150 μM alpha-lipoic acid for four min. The VS1 was discarded and then the fragments were submerged into a second vitrification solution (VS2) containing the same composition of VS1 but 20% EG and 20% DMSO. VS2 was then discarded and each OTC device containing up to four testicular fragments was closed and immersed in NL. After the storage period, the fragments were removed from the NL, kept at room temperature for one min and then immersed at 37 °C in a water bath for 30 s. Samples were warmed by sequentially immersing in solutions of MEM-HEPES supplemented with 3 mg/mL BSA and decreasing concentrations of sucrose. Hematoxylin-eosin staining to analyze the tissue architecture was used. The score scale used was from 0 to 3, classified with a score 0 representing normal morphologically, and 3 were considered a lot of alteration. The histomorphological evaluation of the testicular tissue shows that when evaluating the nuclear alteration (distinction of nucleoli and condensation of nuclei), there are no differences when using slow freezing with respect to the control. However, vitrification presents greater damage (p <0.05). On the other hand, when evaluating the epithelial alteration, we observed that the freezing showed scores statistically equal to the control in variables such as retraction of the basement membrane, formation of gaps and organization of the peritubular cells. The results of the study demonstrated that cryopreservation using the slow freezing method is an excellent tool for the preservation of pubertal testicular tissue.Keywords: cryopreservation, slow freezing, vitrification, testicular tissue, lambs
Procedia PDF Downloads 174491 Characterization of Novel Bi-Directional Promoter from Begomovirus: A Breakthrough in Plant Genomics
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
Procedia PDF Downloads 332490 The New Insight about Interspecies Transmission of Iranian H9N2 Influenza Viruses from Avian to Human
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
Procedia PDF Downloads 449489 MNH-886(Bt.): A Cotton Cultivar (G. Hirsutum L.) for Cultivation in Virus Infested Regions of Pakistan, Having High Seed Cotton Yield and Desirable Fibre Characteristics
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
Procedia PDF Downloads 318488 Biocompatibility and Sensing Ability of Highly Luminescent Synthesized Core-Shell Quantum Dots
Authors: Mohan Singh Mehata, R. K. Ratnesh
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CdSe, CdSe/ZnS, and CdSe/CdS core-shell quantum dots (QDs) of 3-4 nm were developed by using chemical route and following successive ion layer adsorption and reaction (SILAR) methods. The prepared QDs have been examined by using X-ray diffraction, high-resolution electron microscopy and optical spectroscopy. The photoluminescence (PL) quantum yield (QY) of core-shell QDs increases with respect to the core, indicating that the radiative rate increases by the formation of shell around core, as evident by the measurement of PL lifetime. Further, the PL of bovine serum albumin is quenched strongly by the presence of core-shall QDs and follow the Stern-Volmer (S-V) relation, whereas the lifetime does not follow the S-V relation, demonstrating that the observed quenching is predominantly static in nature. Among all the QDs, the CdSe/ZnS QDs shows the least cytotoxicity hence most biocompatibility.Keywords: biocompatibility, core-shell quantum dots, photoluminescence and lifetime, sensing ability
Procedia PDF Downloads 236487 Molecularly Imprinted Nanoparticles (MIP NPs) as Non-Animal Antibodies Substitutes for Detection of Viruses
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
Procedia PDF Downloads 362486 Immunoglobulins and Importance in Ruminants
Authors: M. Akoz, O. B. Citil, I. Aydin
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Colostrum secreted by the mammary glands after birth in the early days, a high proportion of fat, protein and ash containing a secretion containing low amounts of casein and lactose. Especially immunoglobulins contain high proportions. Maternal immunoglobulins own immune system to protect the newborn against neonatal disease until development are very important matter. However, colostrum is transferred to the offspring due to placental barrier in ruminants. Immunoglobulins are absorbed through the intestinal epithelium but absorption can vary under the influence of some factors. These factors are among the priority ones taking colostrum first time, amount, concentration, the metabolic status of the newborn. intestinal absorption of immunoglobulins occurs over the first 24 h high. Absorption from the gut after nine hours, 50% after 24 hours was only 11%. On the other hand pup's digestive system degrade the enzymes after 24 hours immunoglobulins. Bovine colostrum in the composition while basic immune IgG, IgA and IgM are also available. Total IgG in colostrum of ruminants, while in other species is a greater amount in blood serum.Keywords: immunoglobulin, ruminants, colostrum, immune system
Procedia PDF Downloads 268485 Sheep Pox Virus Recombinant Proteins To Develop Subunit Vaccines
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
Procedia PDF Downloads 242484 Development of Peptide Inhibitors against Dengue Virus Infection by in Silico Design
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
Procedia PDF Downloads 357483 No Histological and Biochemical Changes Following Administration of Tenofovir Nanoparticles: Animal Model Study
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
Procedia PDF Downloads 183482 Public Perception and Willingness to Undergo Cosmetic Procedures during COVID-19 Pandemic: A Questionnaire-Based Study Applied to Asymptomatic Individuals
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
Procedia PDF Downloads 182481 Insulin Resistance in Patients with Chronic Hepatitis C Virus Infection: Upper Egypt Experience
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
Procedia PDF Downloads 154480 Green Synthesis of Red-Fluorescent Gold Nanoclusters: Characterization and Application for Breast Cancer Detection
Authors: Agnė Mikalauskaitė, Renata Karpicz, Vitalijus Karabanovas, Arūnas Jagminas
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The use of biocompatible precursors for the synthesis and stabilization of fluorescent gold nanoclusters (NCs) with strong red photoluminescence creates an important link between natural sciences and nanotechnology. Herein, we report the cost-effective synthesis of Au nanoclusters by templating and reduction of chloroauric acid with the cheap amino acid food supplements. This synthesis under the optimized conditions leads to the formation of biocompatible Au NCs having good stability and intense red photoluminescence, peaked at 680 to 705 nm, with a quantum yield (QY) of ≈7% and the average lifetime of up to several µs. The composition and luminescent properties of the obtained NCs were compared with ones formed via well-known bovine serum albumin reduction approach. Our findings implied that synthesized Au NCs tend to accumulate in more tumorigenic breast cancer cells (line MDA-MB-213) and after dialysis can be prospective for bio imagining.Keywords: gold nanoclusters, proteins, materials chemistry, red-photoluminescence, bioimaging
Procedia PDF Downloads 278479 Detection of JC Virus DNA and T-Ag Expression in a Subpopulation of Tunisian Colorectal Carcinomas
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
Procedia PDF Downloads 363