Search results for: Zika virus

Authors: Nino G. Vepkhvadze, Tea Enukidze

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Every year the number of countries around the world face the risk of the spread of infectious diseases that bring significant ecological and social-economic damage. Hence, the importance of food product safety is emphasized that is the issue of interest for many countries. To solve them, it’s necessary to conduct preventive measures against the diseases, have accurate diagnostic results, leadership, and management. The Peste des petits ruminants (PPR) disease is caused by a morbillivirus closely related to the rinderpest virus. PPR is a transboundary disease as it emerges and evolves, considered as one of the top most damaging animal diseases. The disease imposed a serious threat to sheep-breeding when the farms of sheep, goats are significantly growing within the country. In January 2016, PPR was detected in Georgia. Up to present the origin of the virus, the age relationship of affected ruminants and the distribution of PPRV in Georgia remains unclear. Due to the nature of PPR, and breeding practices in the country, reemerging of the disease in Georgia is highly likely. The purpose of the studies is to provide laboratories with efficient tools allowing the early detection of PPR emergence and re-emergences. This study is being accomplished under the Biological Threat Reduction Program project with the support of the Defense Threat Reduction Agency (DTRA). The purpose of the studies is to investigate the samples and identify areas at high risk of the disease. Georgia has a high density of small ruminant herds bred as free-ranging, close to international borders. Kakheti region, Eastern Georgia, will be considered as area of high priority for PPR surveillance. For this reason, in 2019, in Kakheti region investigated n=484 sheep and goat serum and blood samples from the same animals, utilized serology and molecular biology methods. All samples were negative by RT-PCR, and n=6 sheep samples were seropositive by ELISA-Ab. Future efforts will be concentrated in areas where the risk of PPR might be high such as international bordering regions of Georgia. For diagnostics, it is important to integrate the PPRV knowledge with epidemiological data. Based on these diagnostics, the relevant agencies will be able to control the disease surveillance.

Keywords: animal disease, especially dangerous pathogen, laboratory diagnostics, virus

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Authors: Zunnu Raen Akhtar

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Cotton Leaf Curl Disease (CLCuD) is from Gemini virus and is transmitted through whiteflies in cotton. Transgenic cotton containing Antisense Coat Protein (ACP) has been found to show better results against CLCuD in cotton. In current research, Antisense Coat Protein was inserted in cotton plants to observe resistance developed in the cotton plants against CLCuD. T1 generation of plants were observed for its expression in plants. Tests were carried out to observe the expression of Antisense Coat Protein using Polymerase Chain Reaction (PCR) technique and by southern blotting. Whiteflies showing positive Cotton Leaf Curl Virus (CLCV) were reared and released in bioassay on ACP expressing cotton plants under laboratory as well as confined semi-field conditions. Results confirmed the expression of AC protein in PCR and southern blotting. Further laboratory results showed that cotton plants expressing AC protein showed rare incidence of CLCuD infection as compared to control. In the confined semi-field, similar results were observed in AC protein expressing cotton as compared to control. These results explicitly show that ACP can help to tackle the CLCuD issue in the future and further studies on biochemical processes involved in these plants and effects of ACP induction on non-target organisms should also be studied for eco-system.

Keywords: cotton, white flies, antisense coat protein, CLCV

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Authors: Rizky Kusuma Cahyani

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Hepatitis B is a liver inflammatory disease caused by hepatitis B virus (HBV). This infection can be prevented by vaccination which contains HBV surface protein (sHBsAg). However, vaccine supply is limited. Several attempts have been conducted to produce local sHBsAg. However, the purity degree and protein yield are still inadequate. Therefore optimization of HBsAg purification steps is required to obtain high yield with better purification fold. In this study, optimization of purification was done in 2 steps, precipitation using variation of NaCl concentration (0,3 M; 0,5 M; 0,7 M) and PEG (3%, 5%, 7%); ion exchange chromatography (IEC) using NaCl 300-500 mM elution buffer concentration.To determine HBsAg protein, bicinchoninic acid assay (BCA) and enzyme-linked immunosorbent assay (ELISA) was used in this study. Visualization of HBsAg protein was done by SDS-PAGE analysis. Based on quantitative analysis, optimal condition at precipitation step was given 0,3 M NaCl and PEG 3%, while in ion exchange chromatography step, the optimum condition when protein eluted with NaCl 500 mM. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis indicates that the presence of protein HBsAg with a molecular weight of 25 kDa (monomer) and 50 kDa (dimer). The optimum condition for purification of sHBsAg produced in Pichia pastoris gave a yield of 47% and purification fold 17x so that it would increase the production of hepatitis B vaccine to be more optimal.

Keywords: hepatitis B virus, HBsAg, hepatitis B surface antigen, Pichia pastoris, purification

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Authors: Taimur Khan, Zakirullah, Muhammad Shahab

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The SARS-CoV-2 variant BA.2.86 (Omicron) has emerged with unique mutations that may increase its transmission and infectivity. This study investigates how these mutations alter the Omicron receptor-binding domain's interaction network and dynamic properties (RBD) compared to the wild-type virus, focusing on its binding affinity to the human ACE2 (hACE2) receptor. Protein-protein docking and all-atom molecular dynamics simulations were used to analyze structural and dynamic differences. Despite the structural similarity to the wild-type virus, the Omicron variant exhibits a distinct interaction network involving new residues that enhance its binding capacity. The dynamic analysis reveals increased flexibility in the RBD, particularly in loop regions crucial for hACE2 interaction. Mutations significantly alter the secondary structure, leading to greater flexibility and conformational adaptability compared to the wild type. Binding free energy calculations confirm that the Omicron RBD has a higher binding affinity (-70.47 kcal/mol) to hACE2 than the wild-type RBD (-61.38 kcal/mol). These results suggest that the altered interaction network and enhanced dynamics of the Omicron variant contribute to its increased infectivity, providing insights for the development of targeted therapeutics and vaccines.

Keywords: SARS-CoV-2, molecular dynamic simulation, receptor binding domain, vaccine

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Authors: Kalpana Poudel-Tandukar

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Background: Human Immunodeficiency Virus (HIV) infection has been frequently associated with vitamin D deficiency and depression. Vitamin D deficiency increases the risk of depression in people without HIV. We assessed the cross-sectional and prospective associations between serum concentrations of 25-hydroxyvitamin D (25[OH]D) and depression in a HIV-positive people. Methods: A survey was conducted among 316 HIV-positive people aged 20-60 years residing in Kathmandu, Nepal for a cross-sectional association at baseline, and among 184 participants without depressive symptoms at baseline who responded to both baseline (2010) and follow-up (2011) surveys for prospective association. The competitive protein-binding assay was used to measure 25(OH)D levels and the Beck Depression Inventory-Ia method was used to measure depression, with cut off score 20 or higher. Relationships were assessed using multiple logistic regression analysis with adjustment of potential confounders. Results: The proportion of participants with 25(OH)D level of <20ng/mL, 20-30ng/mL, and >30ng/mL were 83.2%, 15.5%, and 1.3%, respectively. Only four participants with 25(OH)D level of >30ng/mL were excluded in the further analysis. The mean 25(OH)D level in men and women were 15.0ng/mL and 14.4ng/mL, respectively. Twenty six percent of participants (men:23%; women:29%) were depressed. Participants with 25(OH)D level of < 20 ng/mL had a 1.4 fold higher odds of depression in a cross-sectional and 1.3 fold higher odds of depression after 18 months of baseline compared to those with 25(OH)D level of 20-30ng/mL (p=0.40 and p=0.78, respectively). Conclusion: Vitamin D may not have significant impact against depression among HIV-positive people with 25(OH)D level below normal ( > 30ng/mL).

Keywords: depression, HIV, Nepal, vitamin D

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Authors: Ren-Jye Lin, Li-Hsiung Lin, Bing-Cheng Liu, Ching-Len Liao

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The human zinc finger protein 36-like protein family, containing zinc finger protein 36-like 1 (ZFP36L1) and zinc finger protein 36-like 2 (ZFP36L2), belongs to CCCH-type zinc-finger protein identified as an RNA-binding protein that participates in controlling posttranscriptional regulation via RNA decay pathways. Recently, we demonstrated that human ZFP36L1 showed potent antiviral activity against flavivirus Infection by both 5´-3´ XRN1 and 3´-5´RNA-exosome RNA decay pathways (Journal of Virology 2022 Jan 12;96(1): e0166521). However, another zinc finger protein 36-like protein member, ZFP36L2, in the host defense response against flaviviruses has yet to be addressed. Here, we also demonstrate that ZFP36L2 functions as a host innate defender against flaviviruses, including Japanese encephalitis virus (JEV) and dengue virus (DENV). Overexpression of ZFP36L2 reduced JEV and DENV infection, and ZFP36L2 knockdown significantly promoted viral replication. Distinct from the antiviral mechanism of ZFP36L1, ZFP36L2 inhibits flavivirus infection by only a 5´-3´ XRN1-mediated RNA decay pathway but not the 3´-5´RNA-exosome RNA decay pathway. Human ZFP36L1 and ZFP36L2 can restrict flavivirus replication by directly binding and destabilizing viral RNA. Thus, for the first time, human zinc finger protein 36-like family members, ZFP36L1 and ZFP36L2, are identified as host antiviral factors that can bind and degrade flavivirus viral RNA by diverse antiviral mechanisms.

Keywords: ZFP36L1, ZFP36L2, 5'-3' exonuclease XRN1, antiviral mechansim

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Authors: Bagul Abhijeet

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Background: Client-Server model is the backbone of today’s internet communication. In which normal user can not have control over particular website or server? By using the same processing model one can have unauthorized access to particular server. In this paper, we discussed about application scenario of hacking for simple website or server consist of unauthorized way to access the server database. This application emerges to autonomously take direct access of simple website or server and retrieve all essential information maintain by administrator. In this system, IP address of server given as input to retrieve user-id and password of server. This leads to breaking administrative security of server and acquires the control of server database. Whereas virus helps to escape from server security by crashing the whole server. Objective: To control malicious attack and preventing all government website, and also find out illegal work to do hackers activity. Results: After implementing different hacking as well as non-hacking techniques, this system hacks simple web sites with normal security credentials. It provides access to server database and allow attacker to perform database operations from client machine. Above Figure shows the experimental result of this application upon different servers and provides satisfactory results as required. Conclusion: In this paper, we have presented a to view to hack the server which include some hacking as well as non-hacking methods. These algorithms and methods provide efficient way to hack server database. By breaking the network security allow to introduce new and better security framework. The terms “Hacking” not only consider for its illegal activities but also it should be use for strengthen our global network.

Keywords: Hacking, Vulnerabilities, Dummy request, Virus, Server monitoring

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Authors: Mohamad Ammar Ayass, Natalya Griko, Victor Pashkov, Wanying Cao, Kevin Zhu, Jin Zhang, Lina Abi Mosleh

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Respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent for coronavirus disease 2019 (COVID-19). Early evidence pointed at the angiotensin-converting enzyme 2 (ACE-2) expressed on the epithelial cells of the lung as the main entry point of SARS-CoV-2 into the cells. The viral entry is mediated by the binding of the Receptor Binding Domain (RBD) of the spike protein that is expressed on the surface of the virus to the ACE-2 receptor. As the number of SARS-CoV-2 variants continues to increase, mutations arising in the RBD of SARS-CoV-2 may lead to the ineffectiveness of RBD targeted neutralizing antibodies. To address this limitation, the objective of this study is to develop a combination of aptamers that target different regions of the RBD, preventing the binding of the spike protein to ACE-2 receptor and subsequent viral entry and replication. A safe and innovative biomedical tool was developed to inhibit viral infection and reduce the harms of COVID-19. In the present study, DNA aptamers were developed against a recombinant trimer S protein using the Systematic Evolution of Ligands by Exponential enrichment (SELEX). Negative selection was introduced at round number 7 to select for aptamers that bind specifically to the RBD domain. A series of 9 aptamers (ADI2010, ADI2011, ADI201L, ADI203L, ADI205L, ADIR68, ADIR74, ADIR80, ADIR83) were selected and characterized with high binding affinity and specificity to the RBD of the spike protein. Aptamers (ADI25, ADI2009, ADI203L) were able to bind and pull down endogenous spike protein expressed on the surface of SARS-CoV-2 virus in COVID-19 positive patient samples and determined by liquid chromatography- tandem mass spectrometry analysis (LC-MS/MS). LC-MS/MS data confirmed that aptamers can bind to the RBD of the spike protein. Furthermore, results indicated that the combination of the 9 best aptamers inhibited the binding of the purified trimer spike protein to the ACE-2 receptor found on the surface of Vero E6 cells. In the same experiment, the combined aptamers displayed a better neutralizing effect than antibodies. The data suggests that the selected aptamers could be used in therapy to neutralize the effect of the SARS-CoV-2 virus by inhibiting the interaction between the RBD and ACE-2 receptor, preventing viral entry into target cells and therefore blocking viral replication.

Keywords: aptamer, ACE-2 receptor, binding inhibitor, COVID-19, spike protein, SARS-CoV-2, treatment

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Authors: Aghasadeghi MR., Bahramali G., Sadat SM., Sadeghi SA., Yousefi M., Khodaei K., Ghorbani M., Sadat Larijani M.

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Background:The emerging COVID-19 pandemic is a serious concernfor the public health worldwide. Despite the many mutations in the virus genome, it is important to find an effective vaccine against viral mutations. Therefore, in current study, we aimed at immunoinformatic evaluation of the virus proteins immunogenicity to design a preventive vaccine candidate, which could elicit humoral and cellular immune responses as well. Methods:Three antigenic regions are included;Spike, Membrane, and Nucleocapsid amino acid sequences were obtained, and possible fusion proteins were assessed andcompared by immunogenicity, structural features, and population coverage. The best fusion protein was also evaluated for MHC-I and MHC-II T-cell epitopes and the linear and conformational B-cell epitopes. Results: Among the four predicted models, the truncated Spike protein in fusion with M and N proteins is composed of 24 highly immunogenic human MHC class I and 29 MHC class II, along with 14 B-cell linear and 61 discontinues epitopes. Also, the selected protein has high antigenicity and acceptable population coverage of 82.95% in Iran and 92.51% in Europe. Conclusion: The data indicate that the truncated Spike-M-N SARS-CoV-2form which could be potential targets of neutralizing antibodies. The protein also has the ability to stimulate humoral and cellular immunity. The in silico study provided the fusion protein as a potential preventive vaccine candidate for further in vivo evaluation.

Keywords: SARS-CoV-2, immunoinformatic, protein, vaccine

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Authors: Ammal M. Metwally, Dalia M. Elmosalami, Walaa A. Fouad, Abla G. Khalifa, Lobna A. El Etreby, Mohamed AbdelRahman

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People with hepatitis C are likely to experience psychological distress related to adjustment issues following diagnosis. Objective: The study was conducted to determine the psycho-social stressors accompanying Hepatitis C virus (HCV) chronic infection. The study focused on immediate and later on reactions to being diagnosed as infected HCV patients. Effect of HCV on disruption of patients’ relationships in term of family relationship and friendship, employment and financial status was assessed. The magnitude and causes of the social stigma and its relation to awareness about illness, level of education were also assessed. Methods: During this study the subjective experiences of people having HCV was explored through a designed questionnaire targeted 540 cases; 359 males and 181 females from ten out of 21 National Treatment Reference Centers of National Hepatology and Tropical Medicine Research Institutes of Ministry of Health (MOH) hospitals. The study was conducted along a period of six months from September 2011 to March 2012. Results: The study revealed that the financial problems are the commonest problems faced by 75.5 % of the cases. More than 70% of the cases suffered from immediate sadness versus 67.4% suffered from worry. Social stigma was reported by 13 % of HCV +patients, the majority of which were females. Conclusions: Exploring the psychosocial consequences of HCV infection can act as pressing motivators for behavior change needed for limiting HCV endemicity in Egypt.

Keywords: Egypt, HCV infection, psycho-social adjustment, stigma

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Authors: A. M. John-Otumu, M. M. Rahman, M. C. Onuoha, E. P. Ojonugwa

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A Convolutional Neural Network (CNN) model for predicting Lassa fever was built using Python 3.8.0 programming language, alongside Keras 2.2.4 and TensorFlow 2.6.1 libraries as the development environment in order to reduce the current high risk of Lassa fever in West Africa, particularly in Nigeria. The study was prompted by some major flaws in existing conventional laboratory equipment for diagnosing Lassa fever (RT-PCR), as well as flaws in AI-based techniques that have been used for probing and prognosis of Lassa fever based on literature. There were 15,679 blood smear microscopic image datasets collected in total. The proposed model was trained on 70% of the dataset and tested on 30% of the microscopic images in avoid overfitting. A 3x3x3 convolution filter was also used in the proposed system to extract features from microscopic images. The proposed CNN-based model had a recall value of 96%, a precision value of 93%, an F1 score of 95%, and an accuracy of 94% in predicting and accurately classifying the images into clean or infected samples. Based on empirical evidence from the results of the literature consulted, the proposed model outperformed other existing AI-based techniques evaluated. If properly deployed, the model will assist physicians, medical laboratory scientists, and patients in making accurate diagnoses for Lassa fever cases, allowing the mortality rate due to the Lassa fever virus to be reduced through sound decision-making.

Keywords: artificial intelligence, ANN, blood smear, CNN, deep learning, Lassa fever

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Authors: Alaa A. Shamaun Al-Abboodi, Yunis A. A. Bapeer

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400 one-day-old male broiler chicks (Ross-308) randomly divided to 2 main groups, 1st main group (GA) was feeding basal diet with medium chain fatty acid (MCFA) at rate of 0.15% and divided to four subgroups, 3 subgroups vaccinated with different routes with Newcastle Disease Virus (NDV) and non-vaccinated group. The 2nd main group (GB) feeding basal diet without MCFA and divided the same as 1st main group. The parameters used in this study included: ND antibody titers at 1, 10, 21, 28, 35 and 42 days of age and values of CD4 and CD8 at 1, 20, 30 and 42 days of age. This experiment detected increase in ND antibodies titers in (G1, G2, G3) groups were fed on basal diet MCFA comparing to groups were fed without adding MCFA (G5, G6, G7) and control groups (G4, G8). The results of cellular immune response (CD4 and CD8) T-cells in broiler chicks indicated that there was obviously significant relationship between dietary Fatty Acid (FA) versus the diet without FA on the level of CD4 parameter, for the entire experimental period. The effect of different ages was statistically significant in creating different values of CD4 level, whereas the CD4 level decreases markedly with age. However, analyzing the data of different vaccination methods, oculonasal method of vaccination led to the highest value of CD4 compared with the oral, S/C and control groups. There were statistical differences in CD8 values due to supplementation of FA versus the basal diet and due to the effect of different age periods. As for the age effect, the CD8 value at 20 days of age was significantly higher than at 42 and 30 days.

Keywords: broiler, CD4 and CD8, fatty acids, Newcastle Disease

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Authors: Kitoholi V. Zhimo

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Unlike the cases of cases of homosexuals, haemophilic and or drug users in USA, France, Africa and other countries, in India the first case of HIV/AIDS was detected in heterosexual female sex workers (FSW) in Chennai in 1986. This image played an important role in understanding HIV/AIDS scenario in the country. Similar to popular and dominant metaphors on HIV/AIDS such as ‘gay plague’, ‘new cancer’, ‘lethal disease’, ‘slim disease’, ‘foreign disease’, ‘junkie disease’, etc. around the world, the social construction of the virus was largely attributed to women in India. It was established that women particularly sex workers are ‘carrier’ and ‘transmitter’ of virus and were categorised as High Risk Groups (HRG’s) alongside homosexuals, transgenders and injecting drug users. Recent literature reveals growing rate of HIV infection among housewives since 1997 which revolutionised public health scenario in India. This means shift from high risk group to general public through ‘bridge population’ encompassing long distance truckers and migrant labours who at the expense of their nature of work and mobility comes in contact with HRG’s and transmit the virus to the general public especially women who are confined to the domestic space. As HIV epidemic expands, married women in monogamous relationship/marriage stand highly susceptible to infection with limited control, right and access over their sexual and reproductive health and planning. In context of Nagaland, a small state in North-eastern part of India HIV/AIDS transmission through injecting drug use dominated the early scene of the epidemic. However, paradigm shift occurred with declining trend of HIV prevalence among injecting drug users (IDU’s) over the past years with the introduction of Opioid Substitution Therapy (OST) and easy access/availability of syringes and injecting needles. Reflection on statistical data reveals that out of 36 states and union territories in India, the position of Nagaland in HIV prevalence among IDU’s has significantly dropped down from 6th position in 2003 to 16th position in 2017. The present face of virus in Nagaland is defined by (hetero) sexual mode of transmission which accounts for about 91% of as reported by Nagaland state AIDS control society (NSACS) in 2016 wherein young and married woman were found to be most affected leading to feminization of HIV/AIDS epidemic in the state. Thus, not only is HIV epidemic feminised but emerged victim to domestic violence which is more often accepted as normal part of heterosexual relationship. In the backdrop of these understanding, the present paper based on ethnographic fieldwork explores the plight, lived experiences and images of HIV+ve women with regard to sexual and reproductive rights against the backdrop of patriarchal system in Nagaland.

Keywords: HIV/AIDS, monogamy, Nagaland, sex worker disease, women

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Authors: Eric Berry White

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The paper examines the causality between solid waste pollution and lockdown. Particularly in 2020, the world experiences the takeover of the Corona virus pandemic, and most countries decided to adopt lockdown measure as the best solution to curtail the spread of the virus. On March 20, 2020, the Government of Liberia implemented a curfew that starts from 3:00PM to 6:00AM. This means that no unauthorized person is allowed to be in the streets during this time. In most developing countries, the issue of public waste and environmental hazard pollution tend to have a high effect among the slum communities where there are markets. This research covers 6 slums communities around the two biggest market hubs within Monrovia, and the result shows that the lockdown measure significantly reduced public waste pollution by reducing the movement of marketers in slum communities , where limited educational and sensitization for young people is reflected on their job market exclusion, jobless circle, and youth workforce concentration in informal work market. The study discovered that with public awareness and sensitization with females, solid waste could be reduced by 13 percentage point. But there is no evidence that awareness among male conduce pollution. within affected communities, Despite the impact of the lockdown on food consumption, these results emphasized that with the right monitoring of waste and aware, pollution could be reduce. By understanding these results and implementing the best policy, the paper recommends that dump sites be close at certain hours.

Keywords: lockdown, environmental, pollution, waste

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Authors: Asma Ammar, Nabiha Bouafia , Asma BenCheikh, Mohamed Mahjoub, Olfa Ezzi, Wadiaa Bannour, Radhia Helali, Mansour Njah

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Background: The objective of the present study was to identify factors associated with vaccination against Hepatitis B virus (HBV) among healthcare workers (HWs) in the University Hospital Center (UHC) Farhat Hached Sousse, Tunisia. Methods: We conducted a descriptive cross-sectional study all licensed physicians (n= 206) and a representative sample of paramedical staff (n= 372) exercising at UHC Hached Sousse (Tunisia) during two months (January and February 2014). Data were collected using a self-administered and pre-tested questionnaire, which composed by 21 questions. In order to determinate factors associated with vaccination against hepatitis B among HWs, this questionnaire was based on the Health Belief Model, one of the most classical behavior theories. Logistic regression with the stepwise method of Hosmer and Lemeshow was used to identify the determinants of the use of vaccination against HBV. Results: The response rates were 79.8%. Fifty two percent believe that HBV is frequent in our healthcare units and 60.6% consider it a severe infection. The prevalence of HWs vaccination was 39%, 95% CI [34.49%; 43.5%]. In multivariate analysis, determinants of the use of vaccination against HBV among HWs were young age (p=10-4), male gender (p = 0. 006), high or very high importance accorded to health (p = 0.035), perception membership in a risk group for HBV infection (p = 0.038) and very favorable or favorable opinion about vaccination against HVB (p=10-4). Conclusion: The results of our study should be considered in any strategy for preventing VHB infection in HWs. In the mean time, coverage with standard vaccines should be improved also by supplying complete information on the risks of VHB infection and on the safety and efficacy of vaccination.

Keywords: Hepatitis B virus, healthcare workers, prevalence, vaccination

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Authors: Suwattana Visetnan, Premruethai Supungul, Sureerat Tang, Ikuo Hirono, Anchalee Tassanakajon, Vichien Rimphanitchayakit

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In animals, infection by Gram-negative bacteria and certain viruses activates the Imd signaling pathway wherein the a NF-κB transcription factor, Relish, is a key regulatory protein for the synthesis of antimicrobial proteins. Infection by yellow head virus (YHV) activates the Imd pathway. To investigate the expression of genes involved in YHV infection and under the influence of PmRelish regulation, RNA interference and suppression subtractive hybridization (SSH) are employed. The genes in forward library expressed in shrimp after YHV infection and under the activity of PmRelish were obtained by subtracting the cDNAs from YHV-infected and PmRelish-knockdown shrimp with cDNAs from YHV-infected shrimp. Opposite subtraction gave a reverse library whereby an alternative set of genes under YHV infection and no PmRelish expression was obtained. Sequencing of 252 and 99 cDNA clones from the respective forward and reverse libraries were done and annotated through blast search against the GenBank sequences. Genes involved in defense and homeostasis were abundant in both libraries, 31% and 23% in the forward and reverse libraries, respectively. They were predominantly antimicrobial proteins, proteinases and proteinase inhibitors. The expression of antimicrobial protein genes, ALFPm3, crustinPm1, penaeidin3 and penaeidin5 were tested under PmRelish silencing and Gram-negative bacterium V. harveyi infection. Together with the results previously reported, the expression of penaeidin5 and also penaeidin3 but not ALFPm3 and crustinPm1 were under the regulation of PmRelish in the Imd pathway.

Keywords: relish, yellow head virus, penaeus monodon, antimicrobial proteins

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Authors: Nidhi S. Chandra, Veena Agrawal, Debprasad Chattopadhyay

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Background: Hepatitis E virus (HEV) is a major cause of acute viral hepatitis in developing countries. It spreads feco orally mainly due to contamination of drinking water by sewage. There is limited data on the genotypic comparison of HEV isolates from sewage water and humans. The aim of this study was to identify genotype and conduct phylogenetic analysis of HEV isolates from sewage water and humans. Materials and Methods: 14 sewage water and 60 serum samples from acute sporadic hepatitis E cases (negative for hepatitis A, B, C) were tested for HEV-RNA by nested polymerase chain reaction (RTnPCR) using primers designed with in RdRp (RNA dependent RNA polymerase) region of open reading frame-1 (ORF-1). Sequencing was done by ABI prism 310. The sequences (343 nucleotides) were compared with each other and were aligned with previously reported HEV sequences obtained from GeneBank, using Clustal W software. A Phylogenetic tree was constructed by using PHYLIP version 3.67 software. Results: HEV-RNA was detected in 49/ 60 (81.67%) serum and 5/14 (35.71%) sewage samples. The sequences obtained from 17 serums and 2 sewage specimens belonged to genotype I with 85% similarity and clustering with previously reported human HEV sequences from India. HEV isolates from human and sewage in North West India are genetically closely related to each other. Conclusion: These finding suggest that sewage acts as reservoir of HEV. Therefore it is important that measures are taken for proper waste disposal and treatment of drinking water to prevent outbreaks and epidemics due to HEV.

Keywords: hepatitis E virus, nested polymerase chain reaction, open reading frame-1, nucleotidies

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Authors: Yung-Feng Yen, I-an Jen, Yi-Ming Arthur Chen

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The molecular mimicry between human immunodeficiency virus (HIV) protein and red blood cell (RBC) antigens could induce the production of anti-RBC autoantibodies. However, the association between HIV infection and subsequent development of autoimmune hemolytic anemia (AIHA) remains unclear. This nationwide population-based cohort study aimed to determine the association between incident AIHA and HIV in Taiwan. From 2000–2012, we identified adult people living with HIV/AIDS (PLWHA) from the Taiwan centers for disease control HIV Surveillance System. HIV-infected individuals were defined by positive HIV-1 western blot. Age- and sex-matched controls without HIV infection were selected from the Taiwan National Health Insurance Research Database for comparison. All patients were followed until Dec. 31, 2012, and observed for occurrence of AIHA. Of 171,468 subjects (19,052 PLWHA, 152,416 controls), 30 (0.02%) had incident AIHA during a mean follow-up of 5.45 years, including 23 (0.12%) PLWHA and 7 (0.01%) controls. After adjusting for potential confounders, HIV infection was found to be an independent risk factor of incident AIHA (adjusted hazard ratio [AHR], 20.9; 95% confidence interval [CI], 8.34-52.3). Moreover, PLWHA receiving HAART were more likely to develop AIHA than those not receiving HAART (AHR, 10.8; 95% CI, 2.90-40.1). Additionally, the risk of AIHA was significantly increased in those taking efavirenz (AHR, 3.15; 95% CI, 1.18-8.43) or atazanavir (AHR, 6.58; 95% CI, 1.88-22.9) component of the HAART. In conclusion, HIV infection is an independent risk factor for incident AIHA. Clinicians need to be aware of the higher risk of AIHA in PLWHA.

Keywords: autoimmune disease , hemolytic anemia, HIV, highly active antiretroviral treatment

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Authors: Manal Kamel, Sara Maher, Hanan El Baz, Faten Salah, Omar Sayyouh, Zeinab Demerdash

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In the recent COVID 19 pandemic, experts of public health have emphasized testing, tracking infected people, and tracing their contacts as an effective strategy to reduce the spread of the virus. Development of rapid and sensitive diagnostic assays to replace reverse transcription polymerase chain reaction (RT-PCR) is mandatory..Our innovative test strip relying on the application of nanoparticles conjugated to recombinant nanobodies for SARS-COV-2 spike protein (S1) & angiotensin-converting enzyme 2 (that is responsible for the virus entry into host cells) for rapid detection of SARS-COV-2 spike protein (S1) in saliva or sputum specimens. Comparative tests with RT-PCR will be held to estimate the significant effect of using COVID 19 nanobodies for the first time in the development of lateral flow test strip. The SARS-CoV-2 S1 (3 ng of recombinant proteins) was detected by our developed LFIA in saliva specimen of COVID-19 Patients No cross-reaction was detected with Middle East respiratory syndrome coronavirus (MERS-CoV) or SARS- CoV antigens..Our developed system revealed 96 % sensitivity and 100% specificity for saliva samples compared to 89 % and 100% sensitivity and specificity for nasopharyngeal swabs. providing a reliable alternative for the painful and uncomfortable nasopharyngeal swab process and the complexes, time consuming PCR test. An increase in testing compliances to be expected.

Keywords: COVID 19, diagnosis, LFIA, nanobodies, ACE2

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Authors: Suthida Panwong, Jeeraporn Pekkoh, Yingmanee Tragoolpua, Aussara Panya

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Dengue virus (DENV) infection is a major public health problem in many countries, especially in tropical and subtropical countries. DENV infection causes dengue fever that can progress to serious conditions of dengue hemorrhagic fever (DHF) or dengue shock syndrome (DSS), relevant to a high risk of mortality. However, there are no effective treatments available against the manifestation and fatalities. Currently, natural extracts have been widely used for the treatment of infectious diseases due to their safety, non-accumulation in the body, or lower side effects. Chlorella spp. is a microalgae with anti-viral activity, but there is not much report to support its ability to inhibit DENV infection. Thus, this study aimed to investigate the inhibitory effect of ethanolic extract from Chlorella sp. AARL G049, which was explored in Thailand on inhibition of DENV-2 infection. The inhibitory effect on viral infection was assessed using a foci-forming assay (FFA), which revealed that a concentration of 125 µg/mL could inhibit viral infection in Vero cells by 75.45±8.06% when treated at the same time as DENV-2 infection. Moreover, the extract at an equal concentration effectively reduced viral protein synthesis by 90.51±5.48% when assessed in human cell lines using enzyme-linked immunosorbent assay (ELISA). Concordantly, the number of infected cells after treatment was reduced as measured by immunofluorescent assay (IFA). Therefore, the finding of this study supports the potential use of Chlorella sp. extract to suppress DENV infection.

Keywords: viral infection, flavivirus, treatment, natural extract

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Authors: A. Asambe, A. K. B. Sackey, L. B. Tekdek

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A serosurveillance study was conducted to detect the presence of antibodies to African swine fever virus (ASFV) and Classical swine fever virus in pigs sampled from piggeries and Makurdi central slaughter slab in Benue State, Nigeria. 416 pigs from 74 piggeries across 12 LGAs and 44 pigs at the Makurdi central slaughter slab were sampled for serum. The sera collected were analysed using Indirect Enzyme Linked Immunosorbent Assay (ELISA) test kit to test for antibodies to ASFV, while competitive ELISA test kit was used to test for antibodies to CSFV. Of the 416 pigs from piggeries and 44 pigs sampled from the slaughter slab, seven (1.7%) and six (13.6%), respectively, tested positive to ASFV antibodies and was significantly associated (p < 0.0001). Out of the 12 LGAs sampled, Obi LGA had the highest ASFV antibody detection rate of (4.8%) and was significantly associated (p < 0.0001). None of the samples tested positive to CSFV antibodies. The study concluded that antibodies to CSFV were absent in the sampled pigs in piggeries and at the Makurdi central slaughter slab in Benue State, while antibodies to ASFV were present in both locations; hence, the need to keep an eye open for CSF too since both diseases may pose great risk in the study area. Further studies to characterise the ASFV circulating in Benue State and investigate the possible sources is recommended. Routine surveillance to provide a comprehensive and readily accessible data base to plan for the prevention of any fulminating outbreak is also recommended.

Keywords: African swine fever, classical swine fever, piggery, slaughter slab, surveillance

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Authors: Samaneh Farokhirad, Fatemeh Ahmadpoor

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Biological membranes are constantly in contact with various filamentous soft nanostructures that either reside on their surface or are being transported between the cell and its environment. In particular, viral infections are determined by the interaction of viruses (such as filovirus) with cell membranes, membrane protein organization (such as cytoskeletal proteins and actin filament bundles) has been proposed to influence the mechanical properties of lipid membranes, and the adhesion of filamentous nanoparticles influence their delivery yield into target cells or tissues. The goal of this research is to integrate the rapidly increasing but still fragmented experimental observations on the adhesion and self-assembly of nanofilaments (including filoviruses, actin filaments, as well as natural and synthetic nanofilaments) on cell membranes into a general, rigorous, and unified knowledge framework. The global outbreak of the coronavirus disease in 2020, which has persisted for over three years, highlights the crucial role that nanofilamentbased delivery systems play in human health. This work will unravel the role of a unique property of all cell membranes, namely flexoelectricity, and the significance of nanofilaments’ flexibility in the adhesion and self-assembly of nanofilaments on cell membranes. This will be achieved utilizing a set of continuum mechanics, statistical mechanics, and molecular dynamics and Monte Carlo simulations. The findings will help address the societal needs to understand biophysical principles that govern the attachment of filoviruses and flexible nanofilaments onto the living cells and provide guidance on the development of nanofilament-based vaccines for a range of diseases, including infectious diseases and cancer.

Keywords: virus nanofilaments, cell mechanics, computational biophysics, statistical mechanics

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Authors: Yahya Almurtadha, Mukhtar Ghaleb, Ahmed M. Shamsan Saleh

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The world witnessed a fierce attack from the Covid-19 virus, which affected public life socially, economically, healthily and psychologically. The world's governments tried to confront the pandemic by imposing a number of precautionary measures such as general closure, curfews and social distancing. Scientists have also made strenuous efforts to develop an effective vaccine to train the immune system to develop antibodies to combat the virus, thus reducing its symptoms and limiting its spread. Artificial intelligence, along with researchers and medical authorities, has accelerated the vaccine development process through big data processing and simulation. On the other hand, one of the most important negatives of the impact of Covid 19 was the state of anxiety and fear due to the blowout of rumors through social media, which prompted governments to try to reassure the public with the available means. This study aims to proposed using Sentiment Analysis (AKA Opinion Mining) and deep learning as efficient artificial intelligence techniques to work on retrieving the tweets of the public from Twitter and then analyze it automatically to extract their opinions, expression and feelings, negatively or positively, about the symptoms they may feel after vaccination. Sentiment analysis is characterized by its ability to access what the public post in social media within a record time and at a lower cost than traditional means such as questionnaires and interviews, not to mention the accuracy of the information as it comes from what the public expresses voluntarily.

Keywords: deep learning, opinion mining, natural language processing, sentiment analysis

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Authors: Simona Pileviciene

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On 24th January 2014, Lithuania notified two primary cases of African swine fever (ASF) in wild boars. The animals were tested positive for ASF virus (ASFV) genome by real-time PCR at the National Reference Laboratory for ASF in Lithuania (NRL), results were confirmed by the European Union Reference Laboratory for African swine fever (CISA-INIA). Intensive wild and domestic animal monitoring program was started. During the period of 2014-2017 ASF was confirmed in two large commercial pig holding with the highest biosecurity. Pigs were killed and destroyed. Since 2014 ASF outbreak territory from east and south has expanded to the middle of Lithuania. Diagnosis by PCR is one of the highly recommended diagnostic methods by World Organization for Animal Health (OIE) for diagnosis of ASF. The aim of the present study was to compare singleplex real-time PCR assays to a duplex assay allowing the identification of ASF and internal control in a single PCR tube and to compare primers, that target the p72 gene (ASF 250 bp and ASF 75 bp) effectivity. Multiplex real-time PCR assays prove to be less time consuming and cost-efficient and therefore have a high potential to be applied in the routine analysis. It is important to have effective and fast method that allows virus detection at the beginning of disease for wild boar population and in outbreaks for domestic pigs. For experiments, we used reference samples (INIA, Spain), and positive samples from infected animals in Lithuania. Results show 100% sensitivity and specificity.

Keywords: African swine fewer, real-time PCR, wild boar, domestic pig

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Authors: Eyüp S. Akbas, Betul Ayaz, Beyza S. Haksever, Sema Basat

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We aimed to evaluate the relationship between insulin resistance, muscle mass and muscle strength in patients with Hepatitis B virus-related cirrhosis. In our study, there were 65 patients with hepatitis B virus-related cirrhosis in Child A and B group and 65 healthy control individual. Control group was chosen between patients who admitted to the internal medicine clinic and had no pathological values in a routine examination. Muscle mass index was calculated with bioimpedance analysis for both groups to determine muscle strength and muscle mass. Handgrip strength, arm, and calf circumference were measured. In both groups, HOMA-IR was calculated to determine insulin resistance. Homeostatic Model Assessment of Insulin Resistance (HOMA-IR) value was detected 3,47±3,80 in the study group and 1,83±1,20 in control group. There were significant differences between the two groups in arm circumference, fasting insulin, fasting glucose, HOMA-IR, High-density lipoprotein (HDL) and total cholesterol parameters. The correlation coefficient between muscle mass and insulin resistance was statistically insignificant, especially in the study group. In healthy individuals group and all the groups, there wasn’t a correlation between muscle mass and insulin resistance. The upper limit for HOMA-IR was determined as 3,2. In control group, %78,9 of individuals were in HOMA-IR ( < 3.2) group and %21,1 of them were in ( ≥ 3,2) group. In study group, %68,3 of individuals were in HOMA-IR ( < 3,2) group and %31.7 were in HOMA-IR ( ≥ 3,2) group. In our study, we did not find a relationship between muscle mass and insulin resistance in patients with liver cirrhosis. In the study group, we detected a positive relationship between muscle mass, handgrip strength, and calf circumference. We did not find a relationship between insulin resistance and handgrip strength in our study.

Keywords: cirrhosis, hepatitis B, Insulin resistance, muscle mass

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

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Cyperus rotendus rhizomes are used as traditional medicine, including Ayurveda in chronic liver diseases and hepatitis B. We investigated the in vitro hepatoprotective and anti-hepatitis B virus (HBV) potential of Cyperus rotundus rhizome organic and aqueous fractions. Of these, the n-butanol and aqueous fractions showed the most promising, dose-dependent hepatoprotection in DCFH-injured HepG2 cells at 48 h. DCFH-toxicated cells were recovered to about 88% and 96%, upon treatment with n-butanol and aqueous fractions (200 g/ml), respectively compared to DCFH-only treated cells. Further, C. rotundus fractions were tested for anti-HBV activities by measuring the expression levels of viral antigens (HBsAg and HBeAg) in the HepG2.2.15 culture supernatants. At 48 h post-treatment, the ethyl acetate, n-butanol and aqueous fractions showed dose-dependent inhibition wherein at a higher dose (100 g/ml), HBsAg production was reduced to 60.27%, 46.87 and 42.76%, respectively. In a time-course study, HBsAg production was inhibited up to 50% and 40% by ethyl acetate and n-butanol fractions (100 g/ml), respectively on day 5. Three three active fractions were further subjected to time-dependent inhibition of HBeAg expression, an indirect measure of HBV active DNA replication. At day 5 post-treatment, ethyl acetate and n-butanol fractions downregulated HBV replication by 44.14% and 24.70%, respectively. In conclusion, our results showed very promising hepatoprotective and anti-HBV potential of C. rotendus tubers fractions in vitro. Our data could, therefore, provide the basis for the claimed traditional use of C. rotendus for jaundice and hepatitis.

Keywords: anti-hepatitis B, cyperus rotundus, hepatitis B virus, hepatoprotection

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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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