Search results for: viral polymerase

Authors: Aman Ullah Khan, Muhammad Younus, Aqil Ijaz, Muti-Ur-Rehman Khan, Sayyed Aun Muhammad, Asif Idrees, Sanan Raza, Amar Nasir

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Dengue fever and malaria are important vector-borne diseases of public health significance affecting millions of people around the globe. Dengue fever is caused by Dengue virus while malaria is caused by plasmodium protozoan. Generally, the consequences of Malaria are less severe compared to dengue fever. This study was designed to differentiate dengue fever and malaria on the basis of clinical and laboratory findings and to compare the changes in both diseases having different causative agents transmitted by the common vector. A total of 200 patients of dengue viral infection (120 males, 80 females) were included in this prospective descriptive study. The blood samples of the individuals were first screened for malaria by blood smear examination and then the negative samples were tested by anti-dengue IgM strip. The strip positive cases were further screened by IgM capture ELISA and their complete blood count including hemoglobin estimation (Hb), total and differential leukocyte counts (TLC and DLC), erythrocyte sedimentation rate (ESR) and platelet counts were performed. On the basis of the severity of signs and symptoms, dengue virus infected patients were subdivided into dengue fever (DF) and dengue hemorrhagic fever (DHF) comprising 70 and 100 confirmed patients, respectively. On the other hand, 30 patients were found infected with Malaria while overall 120 patients showed thrombocytopenia. The patients of DHF were found to have more leucopenia, raised hemoglobin level and thrombocytopenia < 50,000/µl compared to the patients belonging to DF and malaria. On the basis of the outcomes of the study, it was concluded that patients affected by DF were at a lower risk of undergoing haematological disturbance than suffering from DHF. While, the patients infected by Malaria were found to have no significant change in their blood components.

Keywords: dengue fever, blood, serum, malaria, ELISA

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Authors: Malik Muhammad Arslan, Muneeb Ullah, Dai Shihan, Daniyal Haider, Xiaodong Yang

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Chest X-rays are instrumental in the detection and monitoring of a wide array of diseases, including viral infections such as COVID-19, tuberculosis, pneumonia, lung cancer, and various cardiac and pulmonary conditions. To enhance the accuracy of diagnosis, artificial intelligence (AI) algorithms, particularly deep learning models like Convolutional Neural Networks (CNNs), are employed. However, these deep learning models demand a substantial and varied dataset to attain optimal precision. Generative Adversarial Networks (GANs) can be employed to create new data, thereby supplementing the existing dataset and enhancing the accuracy of deep learning models. Nevertheless, GANs have their limitations, such as issues related to stability, convergence, and the ability to distinguish between authentic and fabricated data. In order to overcome these challenges and advance the detection and classification of CXR normal and abnormal images, this study introduces a distinctive technique known as DCWGAN (Diverse Conditional Wasserstein GAN) for generating synthetic chest X-ray (CXR) images. The study evaluates the effectiveness of this Idiosyncratic DCWGAN technique using the ResNet50 model and compares its results with those obtained using the traditional GAN approach. The findings reveal that the ResNet50 model trained on the DCWGAN-generated dataset outperformed the model trained on the classic GAN-generated dataset. Specifically, the ResNet50 model utilizing DCWGAN synthetic images achieved impressive performance metrics with an accuracy of 0.961, precision of 0.955, recall of 0.970, and F1-Measure of 0.963. These results indicate the promising potential for the early detection of diseases in CXR images using this Inimitable approach.

Keywords: CNN, classification, deep learning, GAN, Resnet50

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Authors: Murali Aarthy, Sanjeev Kumar Singh

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High risk human papillomaviruses are highly associated with the carcinoma of the cervix and the other genital tumors. Cervical cancer develops through the multistep process in which increasingly severe premalignant dysplastic lesions called cervical intraepithelial neoplastic progress to invasive cancer. The oncoprotein E7 of human papillomavirus expressed in the lower epithelial layers drives the cells into S-phase creating an environment conducive for viral genome replication and cell proliferation. The replication of the virus occurs in the terminally differentiating epithelium and requires the activation of cellular DNA replication proteins. To date, no suitable drug molecule is available to treat HPV infection whereas identification of potential drug targets and development of novel anti-HPV chemotherapies with unique mode of actions are expected. Hence, our present study aimed to identify the potential inhibitors analogous to EGCG, a green tea molecule which is considered to be safe to use for mammalian systems. A 3D similarity search on the natural small molecule library from natural product database using EGCG identified 11 potential hits based on their similarity score. The structure based docking strategies were implemented in the potential hits and the key interacting residues of protein with compounds were identified through simulation studies and binding free energy calculations. The conformational changes between the apoprotein and the complex were analyzed with the simulation and the results demonstrated that the dynamical and structural effects observed in the protein were induced by the compounds and indicated the dominance to the oncoprotein. Overall, our study provides the basis for the structural insights of the identified potential hits and EGCG and hence, the analogous compounds identified can be potent inhibitors against the HPV 16 E7 oncoprotein.

Keywords: EGCG, oncoprotein, molecular dynamics simulation, analogues

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Authors: Don Anushka Sandaruwan Elvitigala, P. D. S. U. Wickramasinghe, Jehee Lee

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Cystatins are a large superfamily of proteins which act as reversible inhibitors of cysteine proteases. Papain proteases and cysteine cathepsins are predominant substrates of cystatins. Cystatin superfamily can be further clustered into three groups as Stefins, Cystatins, and Kininogens. Among them, stefines are also known as family 1 cystatins which harbors cystatin Bs and cystatin As. In this study, a homologue of family one cystatins more close to cystatin Bs was identified from Korean black rockfish (Sebastes schlegeli) using a prior constructed cDNA (complementary deoxyribonucleic acid) database and designated as RfCyt1. The full-length cDNA of RfCyt1 consisted of 573 bp, with a coding region of 294 bp. It comprised a 5´-untranslated region (UTR) of 55 bp, and 3´-UTR of 263 bp. The coding sequence encodes a polypeptide consisting of 97 amino acids with a predicted molecular weight of 11kDa and theoretical isoelectric point of 6.3. The RfCyt1 shared homology with other teleosts and vertebrate species and consisted conserved features of cystatin family signature including single cystatin-like domain, cysteine protease inhibitory signature of pentapeptide (QXVXG) consensus sequence and N-terminal two conserved neighboring glycine (⁸GG⁹) residues. As expected, phylogenetic reconstruction developed using the neighbor-joining method showed that RfCyt1 is clustered with the cystatin family 1 members, in which more closely with its teleostan orthologues. An SYBR Green qPCR (quantitative polymerase chain reaction) assay was performed to quantify the RfCytB transcripts in different tissues in healthy and immune stimulated fish. RfCyt1 was ubiquitously expressed in all tissue types of healthy animals with gill and spleen being the highest. Temporal expression of RfCyt1 displayed significant up-regulation upon infection with Aeromonas salmonicida. Recombinantly expressed RfCyt1 showed concentration-dependent papain inhibitory activity. Collectively these findings evidence for detectable protease inhibitory and immunity relevant roles of RfCyt1 in Sebastes schlegeli.

Keywords: Sebastes schlegeli, family 1 cystatin, immune stimulation, expressional modulation

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Authors: Alexandra-Monica Toma

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Internet memes are an element of computer mediated communication and an important part of online culture that combines text and image in order to generate meaning. This term coined by Richard Dawkings refers to more than a mere way to briefly communicate ideas or emotions, thus naming a complex and an intensely perpetuated phenomenon in the virtual environment. This paper approaches memes as a cultural artefact and a virtual trope that mirrors societal concerns and issues, and analyses the pragmatics of their use. Memes have to be analysed in series, usually relating to some image macros, which is proof of the interplay between imitation and creativity in the memes’ writing process. We believe that their potential to become viral relates to three key elements: adaptation to context, reference to a successful meme series, and humour (jokes, irony, sarcasm), with various pragmatic functions. The study also uses the concept of multimodality and stresses how the memes’ text interacts with the image, discussing three types of relations: symmetry, amplification, and contradiction. Moreover, the paper proves that memes could be employed as speech acts with illocutionary force, when the interaction between text and image is enriched through the connection to a specific situation. The features mentioned above are analysed in a corpus that consists of memes related to the COVID-19 pandemic. This corpus shows them to be highly adaptable to context, which helps build the feeling of connection and belonging in an otherwise tremendously fragmented world. Some of them are created based on well-known image macros, and their humour results from an intricate dialogue between texts and contexts. Memes created in relation to the COVID-19 pandemic can be considered speech acts and are often used as such, as proven in the paper. Consequently, this paper tackles the key features of memes, makes a thorough analysis of the memes sociocultural, linguistic, and situational context, and emphasizes their intertextuality, with special accent on their illocutionary potential.

Keywords: context, memes, multimodality, speech acts

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Authors: Abtehal Y. Anaas, Mohd. Nazmi Bin Abd. Manap

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Meat Tenderness is one of the most important factors affecting consumers' assessment of meat quality. Variation in meat tenderness is genetically controlled and varies among breeds, and it is also influenced by environmental factors that can affect its creation during rigor mortis and postmortem. The final postmortem meat tenderization relies on the extent of proteolysis of myofibrillar proteins caused by the endogenous activity of the proteolytic calpain system. This calpain system includes different calcium-dependent cysteine proteases, and an inhibitor, calpastatin. It is widely accepted that in farm animals including chickens, the μ-calpain gene (CAPN1) is a physiological candidate gene for meat tenderness. This study aimed to identify the association of single nucleotide polymorphism (SNP) markers in the CAPN1 gene with the tenderness of chicken breast meat from two Malaysian native and commercial broiler breed crosses. Ten, five months old native chickens and ten, 42 days commercial broilers were collected from the local market and breast muscles were removed two hours after slaughter, packed separately in plastic bags and kept at -20ºC for 24 h. The tenderness phenotype for all chickens’ breast meats was determined by Warner-Bratzler Shear Force (WBSF). Thawing and cooking losses were also measured in the same breast samples before using in WBSF determination. Polymerase chain reaction (PCR) was used to identify the previously reported C7198A and G9950A SNPs in the CAPN1 gene and assess their associations with meat tenderness in the two breeds. The broiler breast meat showed lower shear force values and lower thawing loss rates than the native chickens (p<0.05), whereas there were similar in the rates of cooking loss. The study confirms some previous results that the markers CAPN1 C7198A and G9950A were not significantly associated with the variation in meat tenderness in chickens. Therefore, further study is needed to confirm the functional molecular mechanism of these SNPs and evaluate their associations in different chicken populations.

Keywords: CAPNl, chicken, meat tenderness, meat quality, SNPs

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Authors: Mohd Izzat Md. Nor, Norfazlina Jaffar, Noor Zaitulakma Md. Zain, Nur Izyanti Mohd Suppian, Subhashini Balakrishnan, Geetha Kandavello

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During the Coronavirus (COVID-19) pandemic, Malaysia has been focusing on building herd immunity by introducing vaccination programs into the community. Hospital Standard Operating Procedures (SOP) were developed to prevent inpatient transmission. Objective: In this study, we focus on the positivity rate of inpatient Person Under Surveillance (PUS) becoming COVID-19 positive and compare this to the National rate in order to see the outcomes of the patient who becomes COVID-19 positive in relation to their vaccination status. Methodology: This is a retrospective observational study carried out from 1 January until 30 March 2022 in Institut Jantung Negara (IJN). There were 5,255 patients admitted during the time of this study. Pre-admission Polymerase Chain Reaction (PCR) swab was done for all patients. Patients with positive PCR on pre-admission screening were excluded. The patient who had exposure to COVID-19-positive staff or patients during hospitalization was defined as PUS and were quarantined and monitored for potential COVID-19 infection. Their frequency and risk of exposure (WHO definition) were recorded. A repeat PCR swab was done for PUS patients that have clinical deterioration with or without COVID symptoms and on their last day of quarantine. The severity of COVID-19 infection was defined as category 1-5A. All patients' vaccination status was recorded, and they were divided into three groups: fully immunised, partially immunised, and unvaccinated. We analyzed the positivity rate of PUS patients becoming COVID-positive, outcomes, and correlation with the vaccination status. Result: Total inpatient PUS to patients and staff was 492; only 13 became positive, giving a positivity rate of 2.6%. Eight (62%) had multiple exposures. The majority, 8/13(72.7%), had a high-risk exposure, and the remaining 5 had medium-risk exposure. Four (30.8%) were boostered, 7(53.8%) were fully vaccinated, and 2(15.4%) were partial/unvaccinated. Eight patients were in categories 1-2, whilst 38% were in categories 3-5. Vaccination status did not correlate with COVID-19 Category (P=0.641). One (7.7%) patient died due to COVID-19 complications and sepsis. Conclusion: Within the first quarter of 2022, our institution's positivity rate (2.6%) is significantly lower than the country's (14.4%). High-risk exposure and multiple exposures to positive COVID-19 cases increased the risk of PUS becoming COVID-19 positive despite their underlying vaccination status.

Keywords: COVID-19, boostered, high risk, Malaysia, quarantine, vaccination status

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Authors: N. R. Prasannakumar, M. N. Maruthi

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The whitefly, Bemisia tabaci (Gennadius) (Hemiptera; Aleyrodidae) is a highly polyphagous pest reported to infest over 600 plant hosts globally. About 42 genetic groups/cryptic species of B. tabaci exist in the world on different hosts. The species have variable behaviour with respect to feeding, development and transmission of viral diseases. Feeding on diverse host plants affect both whitefly development and the population of the endosymbionts harboured by the insects. Due to changes in the level of endosymbionts, the virus transmission efficiency by the vector also gets affected. We investigated these interactions on five host plants – egg plant, tomato, beans, okra and cotton - using a single whitefly species Asia 1 infected with three different bacteria Portiera, Wolbachia and Arsenophonus. The Asia 1 transmits the Tomato leaf curl Bangalore virus (ToLCBV) effectively and thus was used in the interaction studies. We found a significant impact of hosts on whitefly growth and development; eggplant was most favourable host, while okra and tomato were least favourable. Among the endosymbiotic bacteria, the titre of Wolbachia was significantly affected by feeding of B. tabaci on different host plants whereas Arsenophonus and Portiera were unaffected. When whitefly fed on ToLCBV-infected tomato plants, the Arsenophonus population was significantly increased, indicating its previously confirmed role in ToLCBV transmission. Further, screening of total proteins of B. tabaci Asia 1 genetic group interacting with ToLCBV coat protein was carried out using Y2H system. Some of the proteins found to be interacting with ToLCBV CP were HSPs 70kDa, GroEL, nucleoproteins, vitellogenins, apolipophorins, lachesins, enolase. The reported protein thus would be the potential targets for novel whitefly control strategies such as RNAi or novel insecticide target sites for sustainable whitefly management after confirmation of genuine proteins.

Keywords: cDNA, whitefly, ToLCBV, endosymbionts, Y2H

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Authors: N. Jait, M. Maamar, H. Khibri, H. Harmouche, N. Mouatssim, W. Ammouri, Z. Tazimezaelek, M. Adnaoui

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Introduction: Uveitis presents with inflammation of the uvea, intraocular, of heterogeneous etiology and presentation. The objective of our study is to describe the clinical and therapeutic characteristics of infectious uveitis in HIV+ and HIV- patients. Patients and Methods: This is a retrospective study conducted at the internal medicine department of CHU Ibn Sina in Rabat over a period of 12 years (2010–2021), collecting 42 cases of infectious uveitis. Results: 42 patients were identified. 34% (14 cases) had acquired immunosuppression (9 cases: 22% had HIV infection and 12% were on chemotherapy), and 66% were immunocompetent. The M/F sex ratio was 1.1. The average age was 39 years old. Uveitis revealed HIV in a single case; 8/9 patients have already been followed, their average viral load is 3.4 log and an average CD4 count is 356/mm³. The revealing functional signs were: ocular redness (27%), decreased visual acuity (63%), visual blurring (40%), ocular pain (18%), scotoma (13%), and headaches (4%). The uveitis was site: anterior (30%), intermediate (6%), posterior (32%), and pan-uveitis (32%); unilateral in 80% of patients and bilateral in 20%. The etiologies of uveitis in HIV+ were: 3 cases of CMV, 2 cases of toxoplasmosis, 1 case of tuberculosis, 1 case of HSV, 1 case of VZV, and 1 case of syphilis. Etiologies of immunocompetent patients: tuberculosis (41%), toxoplasmosis (18%), syphilis (15%), CMV infection (4 cases: 10%), HSV infection (4 cases: 10%) , lepromatous uveitis (1 case: 2%), VZV infection (1 case: 2%), a locoregional infectious cause such as dental abscess (1 case: 2%), and one case of borreliosis (3% ). 50% of tuberculous uveitis was of the pan-uveitis type, 75% of the uveitis by toxoplasmosis was of the posterior type. Uveitis was associated with other pathologies in 2 seropositive cases (cerebral vasculitis, multifocal tuberculosis). A specific treatment was prescribed in all patients. The initial evolution was favorable in 67%, including 12% HIV+. 11% presented relapses of the same seat during uveitis of the toxoplasmic, tuberculous and herpetic type. 47% presented complications, of which 4 patients were HIV+: 3 retinal detachments; 7 Retinal hemorrhages. 6 unilateral blindness (including 2 HIV+ patients). Conclusion: In our series, the etiologies of infectious uveitis differ between HIV+ and HIV- patients. In HIV+ patients most often had toxoplasmosis and CMV, while HIV - patients mainly presented with tuberculosis and toxoplasmosis. The association between HIV and uveitis is undetermined, but HIV infection was an independent risk factor for uveitis.

Keywords: uveitis, HIV, immunosuppression, infection

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

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

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

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Authors: Dmitry Yu. Korulkin, Raissa A. Muzychkina

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Currently to treat infectious diseases bioactive substances of plant origin having fewer side effects than synthetic medicines and medicines similar to natural components of a human body by the structure and action, become very important. One of the groups of secondary metabolites of the plants - alkaloids can be related the number of the most promising sources of medicines of plant origin. Currently, the structure of more than 7500 compounds has been identified. Analyzing the scope of research in the field of chemistry, pharmacology and technology of alkaloids, we can make a conclusion about that there is no system approach during the research of relation structure-activity on different groups of these substances. It is connected not only with a complex structure of their molecules, but also with insufficient information on the nature of their effect on organs, tissues and other targets in organism. The purpose of this research was to identify pharmacophore groups in the structure of alkaloids of endemic Polygonum L. plants growing in Kazakhstan responsible for their antiviral action. To isolate alkaloids pharmacopoeian methods were used. Antiviral activity of alkaloids of Polygonum L. plants was researched in the Institute of Microbiology and Virology of the Ministry of Education and Science of the Republic of Kazakhstan. Virus-inhibiting properties of compounds were studies in experiments with ortho- and paramyxoviruses on the model of chick-embryos. Anti-viral properties were determined using ‘screening test’ method designed to neutralization of a virus at the amount of 100EID50 with set concentrations of medicines. The difference of virus titer compared to control group was deemed as the criterion of antiviral action. It has been established that Polygonum L. alkaloids has high antiviral effect to influenza and parainfluenza viruses. The analysis of correlation of the structure and antiviral activity of alkaloids allowed identifying the main pharmacophore groups, among which the most important are glycosidation, the presence of carbonyl and hydroxyl groups, molecular weight and molecular size.

Keywords: alkaloids, antiviral, bioactive substances, isolation, pharmacophore groups, Polygonum L.

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

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Dengue virus infection is now considered as one of the most important mosquito-borne infection in human. The virus is known to promote vascular permeability, cerebral edema leading to Dengue hemorrhagic fever (DHF) or Dengue shock syndrome (DSS). Dengue infection has known to be endemic in India for over two centuries as a benign and self-limited disease. In the last couple of years, the disease symptoms have changed, manifesting severe secondary complication. So far, Delhi has experienced 12 outbreaks of dengue virus infection since 1997 with the last reported in 2014-15. Without specific antivirals, the case management of high-risk dengue patients entirely relies on supportive care, involving constant monitoring and timely fluid support to prevent hypovolemic shock. Nonetheless, the diverse clinical spectrum of dengue disease, as well as its initial similarity to other viral febrile illnesses, presents a challenge in the early identification of this high-risk group. WHO recommends the use of warning signs to identify high-risk patients, but warning signs generally appear during, or just one day before the development of severe illness, thus, providing only a narrow window for clinical intervention. The ability to predict which patient may develop DHF and DSS may improve the triage and treatment. With the recent discovery of high throughput RNA sequencing allows us to understand the disease progression at the genomic level. Here, we will collate the results of RNA-Sequencing data obtained recently from PBMC of different categories of dengue patients from India and will discuss the possible role of deregulated genes and long non-coding RNAs NEAT1 for development of disease progression.

Keywords: long non-coding RNA (lncRNA), dengue, peripheral blood mononuclear cell (PBMC), nuclear enriched abundant transcript 1 (NEAT1), dengue hemorrhagic fever (DHF), dengue shock syndrome (DSS)

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Authors: Okan Cakir, Okan Tatli

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Acute motor and sensory axonal neuropathy (AMSAN) syndrome usually occurs following a postviral infection in two to four weeks and is a polyneuropathy characterized by axonal and sensorial degeneration as a rare variant of Gullian-Barre syndrome. In our case, we wanted to mention that a rare case of AMSAN Syndrome due to prior surgery. A 61-year-old male case admitted to emergency department with complaints of weakness in feet, numbness and incapability to walk. In his history, it was learned that endovascular aneurysm repair (EVAR) had applied for abdominal aort aneurysm two weeks ago before admission, his complaints had been for a couple of days increasingly and bilaterally, and there had been no infection disease history for four weeks. In physical examination, general status was good, vital signs were stable, and there was a mild paresis in dorsal flexion of feet in bilaterally lower extremities. No nuchal rigidity was determined. Other system examinations were normal. Urea:52 mg/dL (normal range: 15-44 mg/dL), creatinine: 1,05 mg/dL (normal range: 0,81-1,4 mg/dL), potassium: 3,68 mmol/L (normal range: 3,5-5,5 mmol/L), glycaemia: 142 mg/dL, calcium: 9,71 mg/dL (normal range: 8,5-10,5 mg/dL), erythrocyte sedimentation rate (ESR): 74 mm/h (normal range: 0-15 mm/h) were determined in biochemical tests. The case was consulted to neurology department and hospitalized. In performing electromyography, it was reported as a bilateral significant axonal degeneration with sensory-motor polyneuropathy. Normal ranges of glycaemia and protein levels were detected in lumbal punction. Viral markers and bucella, toxoplasma, and rubella markers were in normal range. Intravenous immunoglobulin (IVIG) was applied as a treatment, physical treatment programme was planned and the case discharged from neurology department. In our case, we mentioned that it should be considered polyneuropathy as an alternative diagnosis in cases admitting symptoms like weakness and numbness had a history of prior surgery.

Keywords: AMSAN Syndrome, emergency department, prior surgery, weakness

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Authors: Xiaolian Jiang, Dongling Liu, Kun Liu

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Aims: To examine the prevalence of POST-TRAUMATIC STRESS DISORDER (PTSD) symptoms among Tibetan adolescents in heavily-hit disaster area three years after Yushu earthquake, and to explore the interactions of the psycho-social-genetic risk factors. Methods: This was a three-stage study. Firstly, demographic variables，PTSD Checklist-Civilian Version （PCL-C），the Internality、Powerful other、Chance Scale，（IPC），Coping Style Scale（CSS），and the Social Support Appraisal（SSA）were used to explore the psychosocial factors of PTSD symptoms among adolescent survivors. PCL-C was used to examine the PTSD symptoms among 4072 Tibetan adolescents，and the Structured Clinical Interview for DSM-IV Disorders（SCID）was used by psychiatrists to make the diagnosis precisely. Secondly，a case-control trial was used to explore the relationship between PTSD and gene polymorphisms. 287adolescents diagnosed with PTSD were recruited in study group, and 280 adolescents without PTSD in control group. Polymerase chain reaction-restriction fragment length polymorphism technology（PCR-RFLP）was used to test gene polymorphisms. Thirdly，SPSS 22.0 was used to explore the interactions of the psycho-social-genetic risk factors of PTSD on the basis of the above results. Results and conclusions: 1．The prevalence of PTSD was 9.70%. 2．The predictive psychosocial factors of PTSD included earthquake exposure, support from others, imagine, abreact, tolerant, powerful others and family support. 3．Synergistic interactions between A1 gene of DRD2 TaqIA and the external locus of control, negative coping style, severe earthquake exposure were found. Antagonism interactions between A1 gene of DRD2 TaqIA and poor social support was found. Synergistic interactions between A1/A1 genotype and the external locus of control, negative coping style were found. Synergistic interactions between 12 gene of 5-HTTVNTR and the external locus of control, negative coping style, severe earthquake exposure were found. Synergistic interactions between 12/12 genotype and the external locus of control, negative coping style, severe earthquake exposure were also found.

Keywords: adolescents, earthquake, PTSD, risk factors

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Authors: Nemat Sokhandan Bashir

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Threats from viruses to agricultural crops could be even larger than the losses caused by the other pathogens because, in many cases, the viral infection is latent but crucial from an epidemic point of view. Wild vegetation can be a source of many viruses that eventually find their destiny in crop plants. Although often asymptomatic in wild plants due to adaptation, they can potentially cause serious losses in crops. Therefore, exploring viruses in wild vegetation is very important. Recently, omics have been quite useful for exploring plant viruses from various plant sources, especially wild vegetation. For instance, we have discovered viruses such as Ambrossia asymptomatic virus I (AAV-1) through the application of metagenomics from Oklahoma Prairie Reserve. Accordingly, extracts from randomly-sampled plants are subjected to high speed and ultracentrifugation to separated virus-like particles (VLP), then nucleic acids in the form of DNA or RNA are extracted from such VLPs by treatment with phenol—chloroform and subsequent precipitation by ethanol. The nucleic acid preparations are separately treated with RNAse or DNAse in order to determine the genome component of VLPs. In the case of RNAs, the complementary cDNAs are synthesized before submitting to DNA sequencing. However, for VLPs with DNA contents, the procedure would be relatively straightforward without making cDNA. Because the length of the nucleic acid content of VPLs can be different, various strategies are employed to achieve sequencing. Techniques similar to so-called "chromosome walking" may be used to achieve sequences of long segments. When the nucleotide sequence data were obtained, they were subjected to BLAST analysis to determine the most related previously reported virus sequences. In one case, we determined that the novel virus was AAV-l because the sequence comparison and analysis revealed that the reads were the closest to the Indian citrus ringspot virus (ICRSV). AAV—l had an RNA genome with 7408 nucleotides in length and contained six open reading frames (ORFs). Based on phylogenies inferred from the replicase and coat protein ORFs of the virus, it was placed in the genus Mandarivirus.

Keywords: wild, plant, novel, metagenomics

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Authors: Hanaa M. Abu El Einin, Ahmed T. Sharaf El- Din

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Biomphalaria snails play an integral role in the transmission of Schistosoma mansoni, the causative agent for human schistosomiasis. Two species of Biomphalaria were reported from Egypt, Biomphalaria alexandrina and Biomphalaria glabrata, and later on a hybrid of B. alexandrina and B. glabrata was reported in streams at Nile Delta. All were known to be excellent hosts of S. mansoni. Host-parasite relationship can be viewed in terms of snail susceptibility and parasite infectivity. The objective of this study will highlight the progress that has been made in using molecular approaches to describe the correct identification of snail species that participating in transmission of schistosomiasis, rapid diagnose of infection in addition to susceptibility and resistance type. Snails were identified using of molecular methods involving Randomly Amplified Polymorphic DNA (RAPD), Polymerase Chain Reaction, Restriction Fragment Length Polymorphisms (PCR-RFLP) and Species - specific- PCR. Molecular approaches to diagnose parasite in snails from Egypt: Nested PCR assay and small subunit (SSU) rRNA gene. Also RAPD PCR for study susceptible and resistance phenotype. The results showed that RAPD- PCR, PCR-RFLP and species-specific-PCR techniques were confirmed that: no evidence for the presence of B. glabrata in Egypt, All Biomphalaria snails collected identified as B. alexandrina snail i-e B alexandrinia is a common and no evidence for hybridization with B. glabrata. The adopted specific nested PCR assay revealed much higher sensitivity which enables the detection of S. mansoni infected snails down to 3 days post infection. Nested PCR method for detection of infected snails using S. mansoni fructose -1,6- bisphosphate aldolase (SMALDO) primer, these primers are specific only for S. mansoni and not cross reactive with other schistosomes or molluscan aldolases Nested PCR for such gene is sensitive enough to detect one cercariae. Genetic variations between B. alexandrina strains that are susceptible and resistant to Schistosoma infec¬tion using a RAPD-PCR showed that 39.8% of the examined snails collected from the field were resistant, while 60.2% of these snails showed high infection rates. In conclusion the genetics of the intermediate host plays a more important role in the epidemiological control of schistosomiasis.

Keywords: biomphalaria, molecular differentiation, parasite detection, schistosomiasis

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Authors: Prince Vivek, Vijay K. Bharti, Manishi Mukesh, Ankita Sharma, Om Prakash Chaurasia, Bhuvnesh Kumar

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High performance of endurance in equid requires adaptive changes involving physio-biochemical, and molecular responses in an attempt to regain homeostasis. We hypothesized that the identification of the suitable reference genes might be considered for assessing of endurance related traits in pony at high altitude and may ensure for individuals struggling to potent endurance trait in ponies at high altitude. A total of 12 mares of ponies, Zanskar breed, were divided into three groups, group-A (without load), group-B, (60 Kg) and group-C (80 Kg) on backpack loads were subjected to a load carry protocol, on a steep climb of 4 km uphill, and of gravel, uneven rocky surface track at an altitude of 3292 m to 3500 m (endpoint). Blood was collected before and immediately after the load carry on sodium heparin anticoagulant, and the peripheral blood mononuclear cell was separated for total RNA isolation and thereafter cDNA synthesis. Real time-PCR reactions were carried out to evaluate the mRNAs expression profile of a panel of putative internal control genes (ICGs), related to different functional classes, namely glyceraldehyde 3-phosphate dehydrogenase (GAPDH), β₂ microglobulin (β₂M), β-actin (ACTB), ribosomal protein 18 (RS18), hypoxanthine-guanine phosophoribosyltransferase (HPRT), ubiquitin B (UBB), ribosomal protein L32 (RPL32), transferrin receptor protein (TFRC), succinate dehydrogenase complex subunit A (SDHA) for normalizing the real-time quantitative polymerase chain reaction (qPCR) data of native pony’s. Three different algorithms, geNorm, NormFinder, and BestKeeper software, were used to evaluate the stability of reference genes. The result showed that GAPDH was best stable gene and stability value for the best combination of two genes was observed TFRC and β₂M. In conclusion, the geometric mean of GAPDH, TFRC and β₂M might be used for accurate normalization of transcriptional data for assessing endurance related traits in Zanskar ponies during load carrying.

Keywords: endurance exercise, ubiquitin B (UBB), β₂ microglobulin (β₂M), high altitude, Zanskar ponies, reference gene

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Authors: Houda Haddad, Nolwen Jouvenet, Maxime Chazal, Frédéric Tangy, Amira Zairi

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Zika virus represents the primary cause of infection during pregnancy and can lead to various neurological disorders, such as microcephaly and Guillain-Barré syndrome, affecting both children and adults. This infection is also associated with urological and nephrological problems. So far, evidence of mosquito-borne Zika virus infection has been reported in a total of 89 countries and territories. However, surveillance efforts primarily concentrate on outbreaks that this virus can cause, yet the measures implemented are typically limited. Currently, there are no specific treatments or vaccines designed for the prevention or treatment of Zika virus infection or its associated disease. The development of effective therapeutic agents presents an urgent need. Importantly, an alternative for advancing the discovery of molecules could be dermaseptins, a family of antimicrobial peptides known for their potential antiviral properties. In this study, we carried out the synthesis of dermaseptins and their analogs and subsequently assessed the bioactivity tests against Zika virus (ZIKV PF13) of dermaseptins B2 and S4 and their derivatives. The cytotoxicity of these peptides was investigated on the HMC3 cell line and HeLa cells by CellTiter-Glo® Luminescent Cell Viability Assay. Thereafter, we evaluated the antiviral activity caused by the action of our dermaseptins on the viral envelope using the Fluorescence Activated Cell Sorting (FACS). The cytotoxicity of our molecules was concentration-dependent at microgram concentrations except for dermaseptin B2 and its derivative, which present no toxicity against HeLa and HMC3 cell lines. It was observed that all tested analogs from the S4 family exhibited antiviral activity with low concentrations ranging from 3 to 12.5 μg/mL, unlike the native B2 and its derivative, which increased the infectivity. Pre-incubating of dermaseptins with ZIKV PF13 before infection revealed that these derivatives inhibit the initial stages of virus infection. In summary, these results suggest that dermaseptins could serve as lead structures for the development of potent antiviral agents against Zika virus infections.

Keywords: dermaseptin B2, dermaseptin S4, analogs, zika virus, neurological infections, antiviral activity

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Authors: Wassie Molla, Klaas Frankena, Mart De Jong

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Lumpy skin disease (LSD) is a severe viral disease of cattle, which often occurs in epidemic form. It is caused by lumpy skin disease virus of the genus capripoxvirus of family poxviridae. Mathematical models play important role in the study of infectious diseases epidemiology. They help to explain the dynamics and understand the transmission of an infectious disease within a population. Understanding the transmission dynamics of lumpy skin disease between animals is important for the implementation of effective prevention and control measures against the disease. This study was carried out in central and north-western part of Ethiopia with the objectives to understand LSD outbreak dynamics, quantify the transmission between animals and herds, and estimate the disease reproduction ratio in dominantly crop-livestock mixed and commercial herd types. Field observation and follow-up study were undertaken, and the transmission parameters were estimated based on a SIR epidemic model in which individuals are susceptible (S), infected and infectious (I), and recovered and immune or dead (R) using the final size and generalized linear model methods. The result showed that a higher morbidity was recorded in infected crop-livestock (24.1%) mixed production system herds than infected commercial production (17.5%) system herds whereas mortality was higher in intensive (4.0%) than crop-livestock (1.5%) system and the differences were statistically significant. The transmission rate among animals and between herds were 0.75 and 0.68 per week, respectively in dominantly crop-livestock production system. The transmission study undertaken in dominantly crop-livestock production system highlighted the presence of statistically significant seasonal difference in LSD transmission among animals. The reproduction numbers of LSD in dominantly crop-livestock production system were 1.06 among animals and 1.28 between herds whereas it varies from 1.03 to 1.31 among animals in commercial production system. Though the R estimated for LSD in different production systems at different localities is greater than 1, its magnitude is low implying that the disease can be easily controlled by implementing the appropriate control measures.

Keywords: commercial, crop-livestock, Ethiopia, LSD, reproduction number, transmission

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Authors: Hafiza Javaria Ashraf, Asim Abbasi, Muhammad Hussnain Babar, Muhammad Sufyan

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Brevicoryne brassicae (Aphid) is not only the major biotic constraint of rapeseed crop but also transmits 20 different viral pathogens that cause diseases in crucifers. Aphids cause major losses to rapeseed by stunting growth and yield, with real damage being contamination of harvested heads. The misuse of pesticides has led to tremendous economic losses and hazards to human health and environmental pollution. Thus, newer approaches for pest control are continuously being sought. The naturally occurring, biologically active plant-based products seem to have a prominent role in the development of future commercial pesticides not only for increased productivity but their eco-friendly nature. The present experiment was carried out in Research Area of Ayub Agriculture Research Institute, Faisalabad to check the efficacy of different botanicals against rapeseed aphid. The tested botanicals were, neem seed extract, neem leaf extract, dathora seed extract, kaner leaf extract and aak leaf extract. Insecticide, advantage 20 EC served as the positive control in the experiment. Data was recorded before and after 1, 3 and 7 days of treatment application. The results of the experiment revealed that neem seed extract exhibited maximum mortality (48.42%) followed by dathora (45.54%) and kaner leaf extract (40.29%) after 7 days of treatment application. However minimum mortality i.e. 26.64% was observed in case of aak leaf extract. Advantage encountered maximum mortality i.e. 86.14%. All treatments caused maximum mortality after 7 days of treatment application. In case of pollinators maximum population reduction was observed in case of insecticide (74.29%) while minimum reduction was observed in neem leaf extract (11.57%). Hence it was concluded that unlike insecticides, plant based products can be a better option for regulating pests and conserving beneficial insect fauna.

Keywords: Aphid, mortality, plant based, pollinators

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Authors: Ljiljana Stojković, Manja Zec, Maja Zivkovic, Maja Bundalo, Marija Glibetić, Dragan Alavantić, Aleksandra Stankovic

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Cardiovascular disease (CVD) is associated with alterations in DNA methylation, the latter modulated by dietary polyphenols. The present pilot study (part of the original clinical study registered as NCT02800967 at www.clinicaltrials.gov) aimed to investigate the impact of 4-week daily consumption of polyphenol-rich Aronia melanocarpa juice on Long Interspersed Nucleotide Element-1 (LINE-1) methylation in peripheral blood leukocytes, in subjects (n=34, age of 41.1±6.6 years) at moderate CVD risk, including an increased body mass index, central obesity, high normal blood pressure and/or dyslipidemia. The goal was also to examine whether factors known to affect DNA methylation, such as folate intake levels, MTHFR C677T gene variant, as well as the anthropometric and metabolic parameters, modulated the LINE-1 methylation levels upon consumption of polyphenol-rich Aronia juice. The experimental analysis of LINE-1 methylation was done by the MethyLight method. MTHFR C677T genotypes were determined by the polymerase chain reaction-restriction fragment length polymorphism method. Folate intake was assessed by processing the data from the food frequency questionnaire and repeated 24-hour dietary recalls. Serum lipid profile was determined by using Roche Diagnostics kits. The statistical analyses were performed using the Statistica software package. In women, after vs. before the treatment period, a significant decrease in LINE-1 methylation levels was observed (97.54±1.50% vs. 98.39±0.86%, respectively; P=0.01). The change (after vs. before treatment) in LINE-1 methylation correlated directly with MTHFR 677T allele presence, average daily folate intake and the change in serum low-density lipoprotein cholesterol, while inversely with the change in serum triacylglycerols (R=0.72, R2=0.52, adjusted R2=0.36, P=0.03). The current results imply potential cardioprotective effects of habitual polyphenol-rich Aronia juice consumption achieved through the modifications of DNA methylation pattern in subjects at CVD risk, which should be further confirmed. Hence, the precision nutrition-driven modulations of DNA methylation may become targets for new approaches in the prevention and treatment of CVD.

Keywords: Aronia melanocarpa, cardiovascular risk, LINE-1, methylation, peripheral blood leukocytes, polyphenol

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Authors: Lethukuthula Ngobese, Abin Gupthar, Patrick Govender

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The ability of yeast cell wall-derived mannoproteins (glycoproteins) to positively contribute to oenological properties has been a key factor that stimulates research initiatives into these industrially important glycoproteins. In addition, and from a fundamental research perspective, yeast cell wall glycoproteins are involved in a wide range of biological interactions. To date, and to the best of our knowledge, our understanding of the fine molecular structure of these mannoproteins is fairly limited. Generally, the amino acid sequences of their protein moieties have been established from structural and functional analysis of the genomic sequence of these yeasts whilst far less information is available on the glycosyl moieties of these mannoproteins. A novel strategy was devised in this study that entails the genetic engineering of yeast strains that over-express and release cell wall-associated glycoproteins into the liquid growth medium. To this end, the Flo1p mannoprotein was overexpressed in Saccharomyces cerevisiae laboratory strains bearing a specific deletion in KNR4 and GPI7 genes involved in cell wall biosynthesis that have been previously shown to extracellularly hyper-secrete cell wall-associated glycoproteins. A polymerase chain reaction (PCR) -based cloning strategy was employed to generate transgenic yeast strains in which the native cell wall FLO1 glycoprotein-encoding gene is brought under transcriptional control of the constitutive PGK1 promoter. The modified Helm’s flocculation assay was employed to assess flocculation intensities of a Flo1p over-expressing wild type and deletion mutant as an indirect measure of their abilities to release the desired mannoprotein. The flocculation intensities of the transformed strains were assessed and all the strains showed similar intensities (>98% flocculation). To assess if mannoproteins were released into the growth medium, the supernatant of each strain was subjected to the BCA protein assay and the transformed Δknr4 strain showed a considerable increase in protein levels. This study has the potential to produce mannoproteins in sufficient quantities that may be employed in future investigations to understand their molecular structures and mechanisms of interaction to the benefit of both fundamental and industrial applications.

Keywords: glycoproteins, genetic engineering, flocculation, over-expression

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Authors: Umar Saeed

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Peptidyl-prolyl cis/trans isomerases Par14 and Par17 in humans play crucial roles in diverse cellular processes, including protein folding, chromatin remodeling, DNA binding, ribosome biogenesis, and cell cycle progression. However, the effects of Par14 and Par17 on viral replication have been explored to a limited extent. We first time discovered their influential roles in promoting Hepatitis B Virus replication. In this study, we observed that in the presence of HBx, either Par14 or Par17 could upregulate HBV replication. However, in the absence of HBx, neither Par14 nor Par17 had any effect on replication. Their mechanism of action involves binding to specific motifs within HBc and HBx proteins. Notably, they target the conserved 133Arg-Pro134 (RP) motif of HBc and the 19RP20-28RP29 motifs of HBx. This interaction is fundamental for the stability of HBx, core particles, and HBc. Par14 and Par17 exhibit versatility by binding both outside and inside core particles, thereby facilitating core particle assembly through their participation in HBc dimer-dimer interactions. NAGE and immunoblotting analyses unveiled the binding of Par14/Par17 to core particles. Co-immunoprecipitation experiments further demonstrated the interaction of Par14/Par17 with core particle assembly-defective and dimer-positive HBc-Y132A. It's essential to emphasize that R133 is the key residue in the HBc RP motif that governs their interaction with Par14/Par17. Chromatin immunoprecipitation conducted on HBV-infected cells elucidated the participation of residues S19 and E46/D74 in Par14 and S44 and E71/D99 in Par17 in the recruitment of 133RP134 motif-containing HBc into cccDNA. Depleting PIN4 in liver cell lines results in a significant reduction in cccDNA levels, pgRNA, sgRNAs, HBc, core particle assembly, and HBV DNA synthesis. Notably, parvulin inhibitors like juglone and PiB have proven to be effective in substantially reducing HBV replication. These inhibitors weaken the interaction between HBV core particles and Par14/Par17, underscoring the dynamic nature of this interaction. It's also worth noting that specific Par14/Par17 inhibitors hold promise as potential therapeutic options for chronic hepatitis B.

Keywords: Par14Par17, HBx, HBc, cccDNA, HBV

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Authors: Zeb Hussain, Muhammad Asif Qureshi, Shaheen Sharafat

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Background: Measles is a contagious viral infection common in childhood. Vaccination against measles is included in the expanded program of immunization (EPI). However, and alarmingly, a high mortality rate is observed due to measles infection in Pakistan. Moreover a recent outbreak of measles in various areas of Pakistan further highlights the problem. It is therefore important to investigate measles specific IgG (antibody) levels in our population. Objective: To quantify measles specific IgG antibodies amongst vaccinated children in district Qamber Shahdadkot, Sindh. Methodology: This cross-sectional study was conducted at the Microbiology section of the Dow-Diagnostic-Research-and-Reference-Laboratory (DDRRL), DUHS after Institutional Review Board approval (IRB-516/DUHS/-14) during August-December-2014. A total of 173 participants (residents of district Qamber Shahdadkot, Sindh) aged between 1-5 years were recruited in the study. Blood samples were collected as per standard phlebotomy guidelines. Blood was stored at 4 °C overnight. Samples were subsequently spun at a speed of 10000rpm to separate sera, which were divided into small aliquots to be frozen at -20 °C. Frozen sera were transported to the DDRRL on dry ice. Measles specific IgG (antibody) titers were quantified using enzyme linked immunosorbant assay (ELISA). Results: Blood was collected from a total of 173 individuals ranging between 1-5 years of age. Of these, a total of 88 participants were males and 85 were females. Of the 173 investigated samples, only 53 (30.6%) showed protective IgG titers against measles while 120 (69%) were sero-negative. Measles specific IgG antibodies titers were higher in female participants compared to the males. Conclusion: Our data demonstrate that a substantial percentage of vaccinated children in district Qamber-Shahdadkot did not have protective antibody titres against measles. It is therefore extremely important to investigate measles specific IgG levels in various parts of Pakistan in order to implement appropriate protective measures.

Keywords: sero-surveillance, measles, vaccinated children, Pakistan

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Authors: Ali Ilter Akdag, Pratima Ray

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Background:This acute gastroenteritis viruses such as rotavirus, astrovirus, and adenovirus are mainly responsible for diarrhea in children below < 5 years old. Molecular detection of these viruses is crucially important to the understand development of the effective cure. This study aimed to determine the prevalence of common these viruses in children < 5 years old presented with diarrhea from Lala Lajpat Rai Memorial Medical College (LLRM) centre (Meerut) North India, India Methods: Total 312 fecal samples were collected from diarrheal children duration 3 years: in year 2014 (n = 118), 2015 (n = 128) and 2016 (n = 66) ,< 5 years of age who presented with acute diarrhea at the Lala Lajpat Rai Memorial Medical College (LLRM) centre(Meerut) North India, India. All samples were the first detection by EIA/RT-PCR for rotaviruses, adenovirus and astrovirus. Results: In 312 samples from children with acute diarrhea in sample viral agent was found, rotavirus A was the most frequent virus identified (57 cases; 18.2%), followed by Astrovirus in 28 cases (8.9%), adenovirus in 21 cases (6.7%). Mixed infections were found in 14 cases, all of which presented with acute diarrhea (14/312; 4.48%). Conclusions: These viruses are a major cause of diarrhea in children <5 years old in North India. Rotavirus A is the most common etiological agent, follow by astrovirus. This surveillance is important to vaccine development of the entire population. There is variation detection of virus year wise due to differences in the season of sampling, method of sampling, hygiene condition, socioeconomic level of the entire people, enrolment criteria, and virus detection methods. It was found Astrovirus higher then Rotavirus in 2015, but overall three years study Rotavirus A is mainly responsible for causing severe diarrhea in children <5 years old in North India. It emphasizes the required for cost-effective diagnostic assays for Rotaviruses which would help to determine the disease burden.

Keywords: adenovirus, Astrovirus, hospitalized children, Rotavirus

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Authors: Sergio Alejandro Cuevas, Catherine Etchebest, Fernando Luis Barroso Da Silva

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The flavivirus genus has several organisms responsible for generating various diseases in humans. Special in Brazil, Zika (ZIKV), Dengue (DENV) and Yellow Fever (YFV) viruses have raised great health concerns due to the high number of cases affecting the area during the last years. Diagnostic is still a difficult issue since the clinical symptoms are highly similar. The understanding of their common structural/dynamical and biomolecular interactions features and differences might suggest alternative strategies towards differential serological diagnostics and therapeutic intervention. Due to their immunogenicity, the primary focus of this study was on the ZIKV, DENV and YFV non-structural proteins 1 (NS1) protein. By means of computational studies, we calculated the main physical chemical properties of this protein from different strains that are directly responsible for the biomolecular interactions and, therefore, can be related to the differential infectivity of the strains. We also mapped the electrostatic differences at both the sequence and structural levels for the strains from Uganda to Brazil that could suggest possible molecular mechanisms for the increase of the virulence of ZIKV. It is interesting to note that despite the small changes in the protein sequence due to the high sequence identity among the studied strains, the electrostatic properties are strongly impacted by the pH which also impact on their biomolecular interactions with partners and, consequently, the molecular viral biology. African and Asian strains are distinguishable. Exploring the interfaces used by NS1 to self-associate in different oligomeric states, and to interact with membranes and the antibody, we could map the strategy used by the ZIKV during its evolutionary process. This indicates possible molecular mechanisms that can explain the different immunological response. By the comparison with the known antibody structure available for the West Nile virus, we demonstrated that the antibody would have difficulties to neutralize the NS1 from the Brazilian strain. The present study also opens up perspectives to computationally design high specificity antibodies.

Keywords: zika, biomolecular interactions, electrostatic interactions, molecular mechanisms

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Authors: John Hang Leung, Shyh-Yau Wang, Hei-Tung Yip, Fion, Ho Tsung-chin, Agnes LF Chan

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Objectives: Platinum-resistant ovarian cancer has a short overall survival of 9–12 months and limited treatment options. The combination of immunotherapy and targeted therapy appears to be a promising treatment option for patients with ovarian cancer, particularly to patients with platinum-resistant recurrent ovarian cancer (PRrOC). However, there are no direct head-to-head clinical trials comparing their efficacy and toxicity. We, therefore, used a network to directly and indirectly compare seven newer immunotherapies or targeted therapies combined with chemotherapy in platinum-resistant relapsed ovarian cancer, including antibody-drug conjugates, PD-1 (Programmed death-1) and PD-L1 (Programmed death-ligand 1), PARP (Poly ADP-ribose polymerase) inhibitors, TKIs (Tyrosine kinase inhibitors), and antiangiogenic agents. Methods: We searched PubMed (Public/Publisher MEDLINE), EMBASE (Excerpta Medica Database), and the Cochrane Library electronic databases for phase II and III trials involving PRrOC patients treated with immunotherapy or targeted therapy plus chemotherapy. The quality of included trials was assessed using the GRADE method. The primary outcomes compared were progression-free survival, the secondary outcomes were overall survival and safety. Results: Seven randomized controlled trials involving a total of 2058 PRrOC patients were included in this analysis. Bevacizumab plus chemotherapy showed statistically significant differences in PFS (Progression-free survival) but not OS (Overall survival) for all interested targets and immunotherapy regimens; however, according to the heatmap analysis, bevacizumab plus chemotherapy had a statistically significant risk of ≥grade 3 SAEs (Severe adverse effects), particularly hematological severe adverse events (neutropenia, anemia, leukopenia, and thrombocytopenia). Conclusions: Bevacizumab plus chemotherapy resulted in better PFS as compared with all interested regimens for the treatment of PRrOC. However, statistical differences in SAEs as bevacizumab plus chemotherapy is associated with a greater risk for hematological SAE.

Keywords: platinum-resistant recurrent ovarian cancer, network meta-analysis, immune checkpoint inhibitors, target therapy, antiangiogenic agents

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Authors: Amir Hamed Abd-Elrahman, Mohamed Bessat

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450 fattening cattle and buffaloes aged from 6 to 30 months old were examined clinically to determine patterns of occurrence of hemoparasitic diseases and the efficacy of different anti theilerial drugs. 420 animals examined clinically to determine relation between different outbreak of FMD and LSD in Egypt 2012- 2013 and haemoprotozoal diseases. The clinical pictures of haemoprotozoal diseases are variable, from sever to mild, depending on the endemic situation which governed by frequent previous exposure and tick infestation. B. bigemina is the most common haemoprotozoal diseases in the area of study and the infection rate in a descending manner for B. bigemina, A. marginale and T. annulata were 20%, 9.7% and 6.6% respectively. The species susceptibility of B. bigemina and T. annulata showed a higher incidence in cattle than buffaloes while in A. marginale showed a little difference in cattle and buffaloes susceptibility by 10% and 9.2% respectively. The breed susceptibility of B. bigemina and T. annulata showed a higher incidence in crossbred cattle than native baladi cattle while A. marginale showed a higher incidence in native baladi cattle than crossbred cattle. The maximal infection rates were recorded during summer months. The infection rates of B. bigemina and A. marginale were higher among young animals over 6 months and declined above 2 year old while in T. annulata the infection rates were lower among young animals and increased above 2 year old. The case fatality of T. annulata was higher than A. marginale and B. bigemina. Efficacy of different anti theilerial drugs were studied, cure rate of chlouroquine group and Butalex group were 60% disappearance of schizont in lymph node smear after 9 days and 5 days respectively while cure rate of Oxytetracycline Dihydrate (Alamycine) group 20% with disappearance of schizont in lymph node smear after 14 days. FMD and LSD infection enhancement the occurrence of bovine haemoprotozoal diseases.

Keywords: Babesia bigemina, Anaplasma marginale, Theileria annulata, FMD, LSD, ephemeral fever

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Authors: Hamed Ahari, Behrouz Akbari Adreghani, Vadood Razavilar, Amirali Anvar, Sima Moradi, Hourieh Shalchi

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Considering the ever increasing population and industrialization of the developmental trend of humankind's life, we are no longer able to detect the toxins produced in food products using the traditional techniques. This is due to the fact that the isolation time for food products is not cost-effective and even in most of the cases, the precision in the practical techniques like the bacterial cultivation and other techniques suffer from operator errors or the errors of the mixtures used. Hence with the advent of nanotechnology, the design of selective and smart sensors is one of the greatest industrial revelations of the quality control of food products that in few minutes time, and with a very high precision can identify the volume and toxicity of the bacteria. Methods and Materials: In this technique, based on the bacterial antibody connection to nanoparticle, a sensor was used. In this part of the research, as the basis for absorption for the recognition of bacterial toxin, medium sized silica nanoparticles of 10 nanometer in form of solid powder were utilized with Notrino brand. Then the suspension produced from agent-linked nanosilica which was connected to bacterial antibody was positioned near the samples of distilled water, which were contaminated with Staphylococcus aureus bacterial toxin with the density of 10-3, so that in case any toxin exists in the sample, a connection between toxin antigen and antibody would be formed. Finally, the light absorption related to the connection of antigen to the particle attached antibody was measured using spectrophotometry. The gene of 23S rRNA that is conserved in all Staphylococcus spp., also used as control. The accuracy of the test was monitored by using serial dilution (l0-6) of overnight cell culture of Staphylococcus spp., bacteria (OD600: 0.02 = 107 cell). It showed that the sensitivity of PCR is 10 bacteria per ml of cells within few hours. Result: The results indicate that the sensor detects up to 10-4 density. Additionally, the sensitivity of the sensors was examined after 60 days, the sensor by the 56 days had confirmatory results and started to decrease after those time periods. Conclusions: Comparing practical nano biosensory to conventional methods like that culture and biotechnology methods(such as polymerase chain reaction) is accuracy, sensitiveness and being unique. In the other way, they reduce the time from the hours to the 30 minutes.

Keywords: exotoxin, nanobiosensor, recognition, Staphylococcus aureus

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Authors: Yugander Arra, Florence Auguy, Melissa Stiebner, Sophie Chéron, Michael M. Wudick, Van Schepler-Luu, Sébastien Cunnac, Wolf B. Frommer, Laurence Albar

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Rice yellow mottle virus (RYMV) is one of the most important diseases affecting rice in Africa. The most promising strategy to reduce yield losses is the use of highly resistant varieties. The resistance gene RYMV2 is homolog of the Arabidopsis constitutive expression of pathogenesis related protein-5 (AtCPR5) nucleoporin gene. Resistance alleles are originating from African cultivated rice Oryza glaberrima, rarely cultivated, and are characterized by frameshifts or early stop codons, leading to a non-functional or truncated protein. Rice possesses two paralogs of CPR5 and function of these genes are unclear. Here, we evaluated the role of the two rice candidate nucleoporin paralogs OsCPR5.1 (pathogenesis-related gene 5; RYMV2) and OsCPR5.2 by CRISPR/Cas9 genome editing. Despite striking sequence and structural similarity, only loss-of-function of OsCPR5.1 led to full resistance, while loss-of-function oscpr5.2 mutants remained susceptible. Short N-terminal deletions in OsCPR5.1 also did not lead to resistance. In contrast to Atcpr5 mutants, neither OsCPR5.1 nor OsCPR5.2 knock out mutants showed substantial growth defects. Taken together, the candidate nucleoporin OsCPR5.1, but not its close homolog OsCPR5.2, plays a specific role for the susceptibility to RYMV, possibly by impairing the import of viral RNA or protein into the nucleus. Whereas gene introgression from O. glaberrima to high yielding O. sativa varieties is impaired by strong sterility barriers and the negative impact of linkage drag, genome editing of OsCPR5.1, while maintaining OsCPR5.2 activity, thus provides a promising strategy to generate O. sativa elite lines that are resistant to RYMV.

Keywords: CRISPR Cas9, genome editing, knock out mutant, recessive resistance, rice yellow mottle virus

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