Search results for: viral hemorrhagic fever
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 635

Search results for: viral hemorrhagic fever

245 Antepartum and Postpartum Pulmonary Cryptococcosis: A Case Report and Systematic Review

Authors: Ghadeer M Alkusayer, Adelicia Yu, Pamela Orr

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Study objective: To report a case of postpartum pulmonary cryptococcal infection (CCI) in an otherwise healthy 35-year-old woman. Additionally, the cases of pulmonary cryptococcal infections either in the antepartum or the postpartum period with pregnancy outcomes, were systematically reviwed. Methods: A systematic search of Cochrane Library, MEDLINE, and EMBASE was conducted for peer-reviewed studies without date restrictions, published in English and relating to CCI during pregnancy or postpartum period. Conference press, editorials, opinion pieces and letters were excluded. Two authors independently screened citations and full-text articles, extracted data and assessed study quality. Given the heterogeneity of study designs, a narrative synthesis was conducted. Results: The search identified 128 references, of which 22 case reports and series met the inclusion criteria. This is a total of 29 women (including the current case) . The mean age of the women was 28.3 ± 12.3 years. Nine (31.03%) presented and were diagnosed in the postpartum period. Two (6.90%) of the patients were reported as immunocompromised with HIV. Four maternal deaths (13.79%) were found in this case series with one (4.3%) patient with severe neurological deficits. Four (17.4%) infant deaths were reported. Women primary presentation varied with chest pain 13 (44.82%), headache 10 (35.70%), dyspnea 19 (65.51%), or fever 12 (41.38%). Three studies reported placental pathology positive for C. neoformans. Conclusion: This case of pulmonary cryptococcal infection in the postpartum period is an important addition to the literature of this rare infection in pregnancy. The patient is not immunocompromised. The patient was successfully treated with 4 months of Fluconazole 400 mg and continued to breastfeed the healthy baby.

Keywords: pulmonary cryptococcus, pregnancy, cryptococci , postpartum

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244 An Internet of Things Smart Washroom Framework

Authors: Robin Ratnasingham, Maher Elshakankiri

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This research report will look at how to make a smart washroom to increase public hygiene and cleanliness. The system would use IoT devices to pick up various activities in the washroom and notify the appropriate stakeholders or devices to regulate the condition of the washroom. As more people are required to physically go back to the office or school, ensuring a clean and sanitized washroom is even more important now than before. It would help prevent virus outbreaks and safeguard the organization from shutdowns or slowdowns in their business. A framework of the suggested smart washroom was introduced to help reduce the chances of a virus outbreak. Most organizations outsource renovation or implementation to an external party. Using the smart washroom framework, we looked at vendors that provide smart washroom solutions. There are IoT vendors that cannot match the framework, and there are vendors that can support the framework design. This segment is a niche market, and most of the devices are similar in their basic functions. However, all the vendors have unique characteristics to give them a competitive advantage over the rest of the IoT washroom companies. Ultimately, the organization would need to decide if they want to add IoT devices to enable smart capability or renovate the washroom to create a fluid IoT smart washroom design. The report would introduce an IoT smart washroom framework to help organizations design a cohesive preventive measure network for the daily maintenance routine. The framework is designed to help understand how to manage washroom cleanliness more efficiently and to provide guidance in achieving this goal. The leading result is eliminating potential viral outbreaks that could jeopardize the organization.

Keywords: IoT, smart washroom, public hygiene, cleanliness, virus outbreaks, safeguard

Procedia PDF Downloads 69
243 Investigation of Leptospira Infection in Stray Animals in Thailand: Leptospirosis Risk Reduction in Human

Authors: Ruttayaporn Ngasaman, Saowakon Indouang, Usa Chethanond

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Leptospirosis is a public health concern zoonosis in Thailand. Human and animals are often infected by contact with contaminated water. The infected animals play an important role in leptospira infection for both human and other hosts via urine. In humans, it can cause a wide range of symptoms, some of which may present mild flu-like symptoms including fever, vomiting, and jaundice. Without treatment, Leptospirosis can lead to kidney damage, meningitis, liver failure, respiratory distress, and even death. The prevalence of leptospirosis in stray animals in Thailand is unknown. The aim of this study was to investigate leptospira infection in stray animals including dogs and cats in Songkhla province, Thailand. Total of 434 blood samples were collected from 370 stray dogs and 64 stray cats during the population control program from 2014 to 2018. Screening test using latex agglutination for the detection of antibodies against Leptospira interrogans in serum samples shows 29.26% (127/434) positive. There were 120 positive samples of stray dogs and 7 positive samples of stray cats. Detection by polymerase chain reaction specific to LipL32 gene of Leptospira interrogans showed 1.61% (7/434) positive. Stray cats (5/64) show higher prevalence than stray dogs (2/370). Although active infection was low detected, but seroprevalence was high. This result indicated that stray animals were not active infection during sample collection but they use to get infected or in a latent period of infection. They may act as a reservoir for domestic animals and human in which stay in the same environment. In order to prevent and reduce the risk of leptospira infection in a human, stray animals should be done health checking, vaccination, and disease treatment.

Keywords: leptospirosis, stray animals, risk reduction, Thailand

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242 Antiulcer Potential of Heme Oxygenase-1 Inducers

Authors: Gaweł Magdalena, Lipkowska Anna, Olbert Magdalena, Frąckiewicz Ewelina, Librowski Tadeusz, Nowak Gabriel, Pilc Andrzej

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Heme oxygenase-1 (HO-1), also known as heat shock protein 32 (HSP32), has been shown to be implicated in cytoprotection in various organs. Its activation plays a significant role in acute and chronic inflammation, protecting cells from oxidative injury and apoptosis. This inducible isoform of HO catalyzes the first and rate-limiting step in heme degradation to produce equimolar quantities of biologically active products: carbon monoxide (CO), free iron and biliverdin. CO has been reported to possess anti-apoptotic properties. Moreover, it inhibits the production of proinflammatory cytokines and stimulates the synthesis of the anti-inflammatory interleukin-10 (IL-10), as well as promotes vasodilatation at sites of inflammation. The second product of catalytic HO-1 activity, free cytotoxic iron, is promptly sequestered into the iron storage protein ferritin, which lowers the pro-oxidant state of the cell. The third product, biliverdin, is subsequently converted by biliverdin reductase into the bile pigment bilirubin, the most potent endogenous antioxidant among the constituents of human serum, which modulates immune effector functions and suppresses inflammatory response. Furthermore, being one of the so-called stress proteins, HO-1 adaptively responds to different stressors, such as reactive oxygen species (ROS), inflammatory cytokines and heavy metals and thus protects cells against such conditions as ischemia, hemorrhagic shock, heat shock or hypoxia. It is suggested that pharmacologic modulation of HO-1 may represent an effective strategy for prevention of stress and drug-induced gastrointestinal toxicity. HO-1 is constitutively expressed in normal gastric, intestinal and colonic mucosa and up-regulated during inflammation. It has been proven that HO-1 up-regulated by hemin, heme and cobalt-protoporphyrin ameliorates experimental colitis. In addition, the up-regulation of HO-1 partially explains the mechanism of action of 5-aminosalicylic acid (5-ASA), which is used clinically as an anti-colitis agent. In 2009 Ueda et al. has reported for the first time that mucosal protection by Polaprezinc, a chelate compound of zinc and L-carnosine used as an anti-ulcer drug in Japan, is also attributed to induction of HO-1 in the stomach. Since then, inducers of HO-1 are desired subject of research, as they may constitute therapeutically effective anti-ulcer drugs.

Keywords: heme oxygenase-1, gastric lesions, gastroprotection, Polaprezinc

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241 Estrogen Controls Hepatitis C Virus Entry and Spread through the GPR30 Pathway

Authors: Laura Ulitzky, Dougbeh-Chris Nyan, Manuel M. Lafer, Erica Silberstein, Nicoleta Cehan, Deborah R. Taylor

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Hepatitis C virus (HCV)-associated hepatocellular carcinoma, fibrosis and cirrhosis are more frequent in men and postmenopausal women than in premenopausal women and women receiving hormone replacement therapy, suggesting that β-estradiol (estrogen) plays an innate role in preventing viral infection and liver disease. Estrogen classically acts through nuclear estrogen receptors or, alternatively, through the membrane-bound G-protein-coupled estrogen receptor (GPR30 or GPER). We observed a marked decrease in detectable virus when HCV-infected human hepatoma cells were treated with estrogen. The effect was mimicked by both Tamoxifen (Tam) and G1, a GPR30-specific agonist, and was reversed by the GPR30-specific antagonist, G15. Through GPR30, estrogen-mediated the down-regulation of occludin; a tight junction protein and HCV receptor, by promoting activation of matrix metalloproteinases (MMPs). Activated MMP-9 was secreted in response to estrogen, cleaving occludin in the extracellular Domain D, the motif required for HCV entry and spread. This pathway gives new insight into a novel innate immune pathway and the disparate host-virus responses to HCV demonstrated by the two sexes. Moreover, these data suggest that hormone replacement therapy may have beneficial antiviral properties for HCV-infected postmenopausal women and show promise for new antiviral treatments for both men and women.

Keywords: HCV, estrogen, occludin, MMPs

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240 Kids and COVID-19: They are Winning with Their Immunity

Authors: Husham Bayazed, Fatimah Yousif

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Purpose of Presentation: The infant immune system has a reputation for being weak and underdeveloped when compared to the adult immune system, but the comparison isn’t quite fair. At the start, as the COVID-19 pandemic drags on and evolves, many Pediatricians and kids' parents have been left with renewed questions about the consequences and sequel of infection on children and the steps to be taken if their child has the symptoms of COVID-19 or tests positive. Recent Findings Literature reviews and recent studies revealed that children are better than adults at controlling SARS-CoV-2. There was conflicting evidence on age-related differences in ACE2 expression in the nose and lungs. But scientists who measured the ‘viral load’ in children's upper airways have seen no clear difference between children and adults. Moreover, the hypothesis is that kids might be more exposed to other coronaviruses common cold, with a production of ready protective antibodies to lock on to the pandemic coronavirus. But the evidence suggests that adults also have this immunity too. Strikingly, these ‘cross-reactive’ antibodies don’t offer any special protection. Summary One of the few silver linings of the Covid-19 pandemic is that children are relatively spared. The kid's Innate Immunity is hardly the whole story, the innate immune response against SARS-CoV-2 infection is early initiative calm with low immunological tone to prevent an overactive immunity and with rapidly repair damage to the lungs in contrast to stormy waves in adults. Therefore, Kids are at much lower risk of Covid-19 infection, and they are still winning the battle against Covid-19 with their innate immunity.

Keywords: Covid-19, kids, ACE2 receptors, immunity

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239 The Role of a Novel DEAD-Box Containing Protein in NLRP3 Inflammasome Activation

Authors: Yi-Hui Lai, Chih-Hsiang Yang, Li-Chung Hsu

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The inflammasome is a protein complex that modulates caspase-1 activity, resulting in proteolytic cleavage of proinflammatory cytokines such as IL-1β and IL-18, into their bioactive forms. It has been shown that the inflammasomes play a crucial role in the clearance of pathogenic infection and tissue repair. However, dysregulated inflammasome activation contributes to a wide range of human diseases such as cancers and auto-inflammatory diseases. Yet, regulation of NLRP3 inflammasome activation remains largely unknown. We discovered a novel DEAD box protein, whose biological function has not been reported, not only negatively regulates NLRP3 inflammasome activation by interfering NLRP3 inflammasome assembly and cellular localization but also mitigate pyroptosis upon pathogen evasion. The DEAD-box protein is the first DEAD-box protein gets involved in modulation of the inflammasome activation. In our study, we found that caspase-1 activation and mature IL-1β production were largely enhanced upon LPS challenge in the DEAD box-containing protein- deleted THP-1 macrophages and bone marrow-derived macrophages (BMDMs). In addition, this DEAD box-containing protein migrates from the nucleus to the cytoplasm upon LPS stimulation, which is required for its inhibitory role in NLRP3 inflammasome activation. The DEAD box-containing protein specifically interacted with the LRR motif of NLRP3 via its DEAD domain. Furthermore, due to the crucial role of the NLRP3 LRR domain in the recruitment of NLRP3 to mitochondria and binding to its adaptor ASC, we found that the interaction of NLRP3 and ASC was downregulated in the presence of the DEAD box-containing protein. In addition to the mechanical study, we also found that this DEAD box protein protects host cells from inflammasome-triggered cell death in response to broad-ranging pathogens such as Candida albicans, Streptococcus pneumoniae, etc., involved in nosocomial infections and severe fever shock. Collectively, our results suggest that this novel DEAD box molecule might be a key therapeutic strategy for various infectious diseases.

Keywords: inflammasome, inflammation, innate immunity, pyroptosis

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238 Rabies Surveillance Data Analysis in Addis Ababa, Ethiopia during 2012/13: Retrospective Cross Sectional Study

Authors: Fantu Lombamo Untiso, Sylvia Murphy, Emily Pieracci

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Background: Rabies is a highly fatal viral disease of all warm-blooded animals including human globally. However, effective rabies control program still remains to be a reality and needs to be strengthened. Objective: Reviewing of recorded data and analyzing it to generate information on the status of rabies in Addis Ababa in the year 2012/13. Methods: A retrospective data were used from the Ethiopian Public Health Institute rabies case record book registered in the year 2012/13. Results: Among 1357 suspected rabid animals clinically examined; only 8.84% were positive for rabies. Out of 216 animal brains investigated in the laboratory with Fluorescent Antibody Technique, 55.5% were confirmed rabies positive. Among the laboratory confirmed positive rabies cases, high percentage of the animals came from Yeka (20%) and lower number from Kirkos subcity (3.3%). Out of 1149 humans who came to the institute seeking anti-rabies post-exposure prophylaxis, 85.65% and 7.87% of them were exposed to suspected dogs and cats respectively. 3 human deaths due to rabies were reported in the year after exposure to dog bite of unknown vaccination status. Conclusion: The principal vector of rabies in Addis Ababa is dog. Effective rabies management and control based on confirmed cases and mass-immunization and control of stray dog populations is recommended.

Keywords: Addis Ababa, exposure, rabies, surveillance

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237 Bacteremia Caused by Nontoxigenic Vibrio cholerae in an Immunocompromised Patient in Istanbul, Turkey

Authors: Fatma Koksal Çakirlar, Si̇nem Ozdemir, Selcan Akyol, Revazi̇ye Gulesen, Murat Gunaydin, Nevri̇ye Gonullu, Belkis Levent, Nuri̇ Kiraz

Abstract:

Vibrio cholerae O1 and O139 are the causative agent of epidemic or pandemic cholera. V. cholerae O1 is generally accepted as a non-invasive enterotoxigenic organism causing gastroenteritis of various severities. Non-O1 V. cholerae can cause small outbreaks of diarrhea due to consumption of contaminated food and water. Particularly, the patients with achlorydria have a risk for vibrio infections. There are numerous case reports of bacteremia caused by vibrio in patients with predisposing conditions like cirrhosis, nephrotic syndrome, diabetes, hematologic malignancy, gastrectomy, and AIDS. We described in this study the first case of nontoxigenic, non-01/non-O139 V. cholerae isolated from the blood culture of a 77-year-old female patient with hipertension, diabetes, coronary artery disease, gout and about 9 years ago migrated breast cancer history. The patient with complaints of shortness of breath, fever and malaise admitted to our emergency clinic were evaluated. There was no diarrhea or abdominal symptoms in the patient. No growth in her urine culture, but blood culture (BACTEC 9120 system, Becton Dickinson, USA) was positive for non-01/non-O139 V. cholerae that was identified by conventional methods and Phoenix automated system (BD Diagnostic Systems, Sparks, MD). It does not secrete the cholera toxin. The agglutination test was negative with polyvalent O1 antisera and O139 antiserum. Empirically ceftriaxone was administered to the patient and she was discharged with improvement in general condition. In this study we report bacteremia by non-01/non-O139 V. cholerae that is rare in the worldwide and first in Turkey.

Keywords: bacteremia, blood culture, immunocompromised patient, Non-O1 vibrio cholerae

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236 The Link of the Human Immunodeficiency Virus With the Progression of Multiple Sclerosis Disease

Authors: Sina Mahdavi

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Multiple sclerosis (MS) is a progressive inflammatory autoimmune disease of the CNS that affects the myelination process in the central nervous system (CNS). Complex interactions of various "environmental or infectious" factors may act as triggers in autoimmunity and disease progression. The association between viral infections, especially human immunodeficiency virus (HIV) and MS is one potential cause that is not well understood. This study aims to summarize the available data on human HIV infection in MS disease progression. In this study, the keywords "Multiple sclerosis", "Human immunodeficiency virus ", and "Central nervous system" in the databases PubMed, and Google Scholar between 2017 and 2022 were searched and 15 articles were chosen, studied, and analyzed. Revealed histologic signs of "MS-like illness" in the setting of HIV, which comprised widespread demyelination with reactive astrocytes, foamy macrophages, and perivascular infiltration with inflammatory cells, all of which are compatible with MS lesions. Human immunodeficiency virus causes dysfunction of the immune system, especially characterized by hypergammaglobulinemia and chronic activation of B cells. Activation of B cells leads to increased synthesis of immunoglobulin and finally to an excess of free light chains. Free light chains may be involved in autoimmune responses against neurons. There is a high expression of HIV during the course of MS, which indicates the relationship between HIV and MS, that this virus can play a role in the development of MS by creating an inflammatory state. Therefore, measures to modulate the expression of HIV may be effective in reducing inflammatory processes in demyelinated areas of MS patients.

Keywords: multiple sclerosis, human immunodeficiency virus, central nervous system, autoimmunity

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235 Biopsy Proven Polyoma (BK) Virus in Saudi Kidney Recipients – Prevalence, Clinicopathological Features and Clinico-Pathological Correlations

Authors: Sarah Hamdan Al-Jahdali, Khaled Alsaad, Abdullah Al-Sayyari

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Objectives: To study the prevalence, clinicopathological features, risk factors and outcome of biopsy proven polyoma (BK) virus infection among Saudi kidney transplant recipients and compare them to negative BK virus group. Methods: We retrospectively reviewed the charts of all the patients with biopsy-proven polyoma (BK) virus infection in King Abdulaziz Medical City in Riyadh between 2005 and 2011. The details of clinical presentation, the indication for kidney biopsy, the laboratory findings at presentation, the natural history of the disease, thepathological findings, the prognosis as well as the response to therapy were all recorded. Results: Kidney biopsy was performed in 37 cases of unexplained graft dysfunction. BK virus was found in 10 (27%). Out of those 10, 3 (30%) ended with graft failure. BK virus occurred in all patients who received ATG induction therapy 100% versus 59.3% in the non BK virus patients (p=0.06). Furthermore, the risk of BK virus was much less in those who received acyclovir as an anti-viral prophylaxis as compared to those who did not receive it (p=0.01). Also, patients with BK virus weighed much less (mean 46.7±20.6 Kgs) than those without BK virus at time of transplantation (mean 64.3±12.1). Graft survival was better among deceased donor kidneys compared to living ones (P=0.016) and with older age (P=0.005). Conclusion: Our findings suggest the involvement of ATG induction therapy, the lack of antiviral prophylaxis therapy and lower weight at transplant as significant risk factors for the development of BK virus infection.

Keywords: BKVAN, BKV, kidney transpant, Saudi Arabia

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234 Linkages of Environment with the Health Condition of Poor Women and Children in the Urban Areas of India

Authors: Barsharani Maharana

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India is the country that shelters the largest number of poor. One of the major areas of concern in India is the unsatisfactory situation of the poor in social developmental and health parameters, not only in rural areas which are partly devoid of the facilities but also in the urban areas where the facilities are insufficient to provide services of a satisfactory quality. Objectives: 1) to examine the association between the environmental condition and health condition among poor women in urban areas. 2) to find out the significance of the effect of environment on the child health among the poor children. 3) to present the scenario of poor among highly urbanized and less urbanized states with respect to the health and environment. Data: data from National Family Health survey-3 and census are used to fulfill the objectives. Methodology: In this study, the standard of living condition of people living in urban areas is computed by taking some household characteristics and assets. People possessing low standard of living are considered as poor. Bivariate and multivariate analysis are employed to examine the effect of environment on poor women and children. A geographical information system is used to present the health and environmental condition of poor in highly and less urbanized states. Results: The findings reveal that the poor women who are not accessed to improved source of water, and sanitation facility are facing more health problems. Children who are living in a dirty environment and are not accessed to improved source of drinking water, among them prevalence of diarrhea and fever is found to be high. As well, the health condition of poor in highly urbanized states is dreadful. Policy implications: Government should emphasize on the implementation of programs regarding the improvement in the infrastructural facilities and health care treatment of urban poor.

Keywords: environment, urban poor, health, sanitation

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233 Design and in Slico Study of the Truncated Spike-M-N SARS-CoV-2 as a Novel Effective Vaccine Candidate

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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232 Preparation of Flurbiprofen Derivative for Enhanced Brain Penetration

Authors: Jungkyun Im

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Nonsteroidal anti-inflammatory drugs (NSAIDs) are effective for relieving pain and reducing inflammation. They are nonselective inhibitors of two isoforms of COX, cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2), and thereby inhibiting the production of hormone-like lipid compounds such as, prostaglandins and thromboxanes which cause inflammation, pain, fever, platelet aggregation, etc. In addition, recently there are many research articles reporting the neuroprotective effect of NSAIDs in neurodegenerative diseases, such as Alzheimer’s disease (AD) and Parkinson’s disease (PD). However, the clinical use of NSAIDs in these diseases is limited by low brain distribution. Therefore, in order to assist the in-depth investigation on the pharmaceutical mechanism of flurbiprofen in neuroprotection and to make flurbiprofen a more potent drug to prevent or alleviate neurodegenerative diseases, delivery of flurbiprofen to brain should be effective and sufficient amount of flurbiprofen must penetrate the BBB thus gaining access into the patient’s brain. We have recently developed several types of guanidine-rich molecular carriers with high molecular weights and good water solubility that readily cross the blood-brain barrier (BBB) and display efficient distributions in the mouse brain. The G8 (having eight guanidine groups) molecular carrier based on D-sorbitol was found to be very effective in delivering anticancer drugs to a mouse brain. In the present study, employing the same molecular carrier, we prepared the flurbiprofen conjugate and studied its BBB permeation by mouse tissue distribution study. Flurbiprofen was attached to a molecular carrier with a fluorescein probe and multiple terminal guanidiniums. The conjugate was found to internalize into live cells and readily cross the BBB to enter the mouse brain. Our novel synthetic flurbiprofen conjugate will hopefully delivery NSAIDs into brain, and is therefore applicable to the neurodegenerative diseases treatment or prevention.

Keywords: flurbiprofen, drug delivery, molecular carrier, organic synthesis

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231 Immunoinformatic Design and Evaluation of an Epitope-Based Tetravalent Vaccine against Human Hand, Foot, and Mouth Disease

Authors: Aliyu Maje Bello, Yaowaluck Maprang Roshorm

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Hand, foot, and mouth disease (HFMD) is a highly contagious viral infection affecting mostly infants and children. Although the Enterovirus A71 (EV71) is usually the major causative agent of HFMD, other enteroviruses such as coxsackievirus A16, A10, and A6 are also found in some of the recent outbreaks. The commercially available vaccines have demonstrated their effectiveness against only EV71 infection but no protection against other enteroviruses. To address the limitation of the monovalent EV71 vaccine, the present study thus designed a tetravalent vaccine against the four major enteroviruses causing HFMD and primarily evaluated the designed vaccine using an immunoinformatics approach. The immunogen was designed to contain the EV71 VP1 protein and multiple reported epitopes from all four distinct enteroviruses and thus designated a tetravalent vaccine. The 3D structure of the designed tetravalent vaccine was modeled, refined, and validated. Epitope screening showed the presence of B-cell, CTL, CD4 T cell, and IFN epitopes with vast application among the Asian population. Docking analysis confirmed the stable and strong binding interactions between the immunogen and immune receptor B-cell receptor (BCR). In silico cloning and immune simulation analyses guaranteed high efficiency and sufficient expression of the vaccine candidate in humans. Overall, the promising results obtained from the in-silico studies of the proposed tetravalent vaccine make it a potential candidate worth further experimental validation.

Keywords: enteroviruses, coxsackieviruses, hand foot and mouth disease, immunoinformatics, tetravalent vaccine

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230 Analgesic, Toxicity and Anti-Pyretic Activities of Methanolic Extract from Hyoscyamus albus Leaves in Albinos Rats

Authors: Yahia Massinissa, Henhouda Affaf, Yahia Mouloud

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The aim of this study was to investigate the toxicity; analgesic and anti-pyretic properties of standardized HA methanolic extract (HAMeOH) in vivo. The acute toxicity study was performed on rats while adopting the OECD-420 Guidelines (fixed dose procedure). Assessment of analgesic activity was performed in rats with two analgesic models. One was acetic acid induced writhing response and the other formalin-induced paw licking. The anti-pyretic effect was tested by brewer’s yeast induced fever in rats. For the acute toxicity test, the higher dose administration of 2000 mg/kg bw. of Hyoscyamus albus did not produce any toxic signs or deaths in rats. There were no significant differences (p>0.05) in the body and organ weights between control and treated groups. The (LD50) of Hyoscyamus albus was higher than 2000 g/kg bw. In subacute toxicity study, no mortality and toxic signs were observed with the doses of 100 and 200 mg/kg bw. of extracts of for 28 consecutive days. These analgesic experimental results indicated that HAMeOH (100 mg/kg and 200 mg/kg) decreased the acetic acid-induced writhing responses and HAMeOH (100 mg/kg and 200 mg/kg) decreased the licking time in the second phase of the formalin test. Moreover, in the model of yeast induced elevation of the body temperature HAMeOH showed dose-dependent lowering of the body temperature up to 3h at both the doses these results obtained, were comparable to that of paracetamol. The present findings indicate that the leaves of Hyoscyamus albus L. possess potent analgesic and antipyretic activity.

Keywords: Hyoscyamus albus, methanolic extract, toxicity, analgesic activity, antipyretic activity, formalin test

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229 Clinical Impact of Ultra-Deep Versus Sanger Sequencing Detection of Minority Mutations on the HIV-1 Drug Resistance Genotype Interpretations after Virological Failure

Authors: S. Mohamed, D. Gonzalez, C. Sayada, P. Halfon

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Drug resistance mutations are routinely detected using standard Sanger sequencing, which does not detect minor variants with a frequency below 20%. The impact of detecting minor variants generated by ultra-deep sequencing (UDS) on HIV drug-resistance (DR) interpretations has not yet been studied. Fifty HIV-1 patients who experienced virological failure were included in this retrospective study. The HIV-1 UDS protocol allowed the detection and quantification of HIV-1 protease and reverse transcriptase variants related to genotypes A, B, C, E, F, and G. DeepChek®-HIV simplified DR interpretation software was used to compare Sanger sequencing and UDS. The total time required for the UDS protocol was found to be approximately three times longer than Sanger sequencing with equivalent reagent costs. UDS detected all of the mutations found by population sequencing and identified additional resistance variants in all patients. An analysis of DR revealed a total of 643 and 224 clinically relevant mutations by UDS and Sanger sequencing, respectively. Three resistance mutations with > 20% prevalence were detected solely by UDS: A98S (23%), E138A (21%) and V179I (25%). A significant difference in the DR interpretations for 19 antiretroviral drugs was observed between the UDS and Sanger sequencing methods. Y181C and T215Y were the most frequent mutations associated with interpretation differences. A combination of UDS and DeepChek® software for the interpretation of DR results would help clinicians provide suitable treatments. A cut-off of 1% allowed a better characterisation of the viral population by identifying additional resistance mutations and improving the DR interpretation.

Keywords: HIV-1, ultra-deep sequencing, Sanger sequencing, drug resistance

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228 Systems for Air Renewal Inside Bus Bodies Importance in the Prevention of Disease Transmission

Authors: Giovanni Matheus Rech, Gilberto Zan, Filipe P. Aguiar

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The current pandemic scenario raises questions that many times would have previously gone unnoticed. One of these issues is the quality of the air we breathe in the most diverse environments in which we are inserted in an everyday. It is plausible to suppose that, at times like this, there is apprehension regarding the possibility of contamination by pathological agents such as viruses and bacterias through the airways. However, the renewal of indoor air, combined with a properly sanitized air conditioning system, are important tools for the prevention of viral diseases, as is the case with COVID-19. The bus is an example of an environment where renovation is applied to improve the quality of indoor air, helping to reduce the possibility of spreading pathological agents. Together with other care, such as an alcohol gel dispenser, curtains to separate the passengers, cleaning the environment more frequently, and mandatory use of masks, help to reduce the transmission of pathologies, such as COVID-19. Knowing the reality of a large part of the population regarding the need for public transport, there are standards and devices dedicated to promoting air quality, ensuring greater comfort and safety for users. This paper seeks to present such standards and recommendations to improve the quality of indoor air, as well as the equipment responsible for the renewal of the air in the body of a bus. Experimental measurement of the flow rates of the renewal devices present in the bus body allows quantifying the average volume of external air admitted in each type of body. This way, it was possible to compare, in terms of airflow per person, the values of a bus in relation to a series of other environments, using recommendations for air renewal are described through the Brazilian standard ABNT NBR 16401.

Keywords: air quality, air renewal, buses, Covid-19

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227 Microbial Contaminants in Drinking Water Collected from Different Regions of Kuwait

Authors: Abu Salim Mustafa

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Water plays a major role in maintaining life on earth, but it can also serve as a matrix for pathogenic organisms, posing substantial health threats to humans. Although, outbreaks of diseases attributable to drinking water may not be common in industrialized countries, they still occur and can lead to serious acute, chronic, or sometimes fatal health consequences. The analysis of drinking water samples from different regions of Kuwait was performed in this study for bacterial and viral contaminations. Drinking tap water samples were collected from 15 different locations of the six Kuwait governorates. All samples were analyzed by confocal microscopy for the presence of bacteria. The samples were cultured in vitro to detect cultivable organisms. DNA was isolated from the cultured organisms and the identity of the bacteria was determined by sequencing the bacterial 16S rRNA genes, followed by BLAST analysis in the database of NCBI, USA. RNA was extracted from water samples and analyzed by real-time PCR for the detection of viruses with potential health risks, i.e. Astrovirus, Enterovirus, Norovirus, Rotavirus, and Hepatitis A. Confocal microscopy showed the presence of bacteria in some water samples. The 16S rRNA gene sequencing of culture grown organisms, followed by BLAST analysis, identified the presence of several non-pathogenic bacterial species. However, one sample had Acinetobacter baumannii, which often causes opportunistic infections in immunocompromised people, but none of the studied viruses could be detected in the drinking water samples analyzed. The results indicate that drinking water samples analyzed from various locations in Kuwait are relatively safe for drinking and do not contain many harmful pathogens.

Keywords: drinking water, microbial contaminant, 16S rDNA, Kuwait

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226 The Influence of Amygdalin on Glioblastoma Multiforme Cell Lines

Authors: Sylwia K. Naliwajko, Justyna Moskwa, Patryk Nowakowski, Renata Markiewicz-Zukowska, Krystyna Gromkowska-Kepka, Anna Puscion-Jakubik, Maria H. Borawska

Abstract:

Amygdalin is found in many fruit seeds, including apricot, peach, quince, apples, and almonds. Amygdalin (also named vitamin B17), as well as its sources, are commonly used as an alternative therapy or prevention of cancer. The potential activity of amygdalin is related to its enzymatic degradation to the hydrogen cyanide. Hydrogen cyanide is a toxic substance that causes liver and nerves damage, fever, coma or even death. Amygdalin is much better tolerated after intravenous than oral administration. The aim of this study was to examine the influence of amygdalin on glioblastoma multiforme cell lines. Three glioblastoma multiforme cell lines – U87MG, T98, LN18 were incubated (48 h) with amygdalin in concentrations 100, 250, 500, 1000 and 2000 µg/mL. The MTT (Thiazolyl Blue Tetrazolium Bromide) test and DNA binding test by [3H]-thymidine incorporation were used to determine the anti-proliferative activity of amygdalin. The secretion of metalloproteinases (MMP2 and MMP-9) from U87MG cells was estimated by gelatin zymography. The statistical analysis was performed using Statistica v. 13.0 software. The data was presented as a % of control. Amygdalin did not show significant inhibition of viability of all the glioblastoma cells in concentrations 100, 250, 500, 1000 µg/mL. In 2000 µg/mL there were significant differences compared to the control, but inhibition of viability was less than 20% (more than 80% of control). The average viability of U87MG cells was 92,0±4%, T98G: 85,8±3% and LN18: 94,7±2% of the control. There was no dose-response viability, and IC50 value was not recognized. DNA binding in U87MG cells was not inhibited (109,0±3 % of control). After treatment with amygdalin, we observed significantly increased secretion of MMP2 and MMP9 in U87MG cells (130,3±14% and 112,0±5% of control, respectively). Our results suggest that amygdalin has no anticancer activity in glioblastoma cell lines.

Keywords: amygdalin, anticancer, cell line, glioblastoma

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225 Oral Antibiotics in Trans-Rectal Prostate Biopsy and Its Efficacy to Reduce Infectious Complications: Systematic Review

Authors: Mohand Yaghi, O. Kehinde

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Background: For the diagnosis of prostate cancer Trans-rectal prostate biopsy (TRPB) is used commonly, the procedure is associated with infective complications. There is evidence that antibiotics (ABx) decrease infective events after TRPB, but different regimens are used. Aim: To systematically review different regimens of prophylactic oral antibiotics in TRPB. Design: Medline, Embase, Clinical trials site, and Cochrane library were searched, experts were consulted about relevant studies. Randomized clinical trials (RCT) conducted in the last twenty years, which investigated different oral antibiotic regimens in TRPB, and compared their efficacy to reduce infectious complications were analyzed. Measurements: Primary outcomes were bacteriuria, urinary tract infection (UTI), fever, bacteremia, sepsis. Secondary outcomes were hospitalization rate, and the prevalence of ABx-resistant bacteria. Results: Nine trials were eligible with 3012 patients. Antibiotics prevented bacteriuria (3.5% vs. 9.88%), UTI (4.46% vs. 9.75%), and hospitalization (0.21% vs. 2.13%) significantly in comparison with placebo or no treatment. No significant difference was found in all outcomes of the review between the single dose regimen and the 3 days. The single dose regimen was as effective as the multiple dose except in Bacteriuria (6.75% vs. 3.25%), and the prevalence of ABx-resistant bacteria (1.57% vs. 0.27%). Quinolones reduced only UTI significantly in comparison with other antibiotics. Lastly, Ciprofloxacin is the best Quinolone to prevent UTI, and hospitalization. Conclusion: it is essential to prescribe prophylactic Antibiotics in TRPB. No conclusive evidence could be claimed about the superiority of the multiple or the 3 days regimens to the single dose regimen. Unexpectedly, ABx-resistant bacteria was identified more often in the single dose cohorts.

Keywords: infection, prostate cancer, sepsis, TRPB

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224 Hemolytic Anemia Monitored After Post-COVID-19 Infection: Changes Related to General Blood Parameters

Authors: Akbarov Elbek Elmurodovich

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Introduction: We are analyzing the topic of hemolytic anemia observed in patients after COVID-19 infection. The purpose of this research is to investigate the development of hemolytic anemia, identify its causes, and study treatment methods. Objective and Task: The goal of our research is to analyze the changes in blood occurring after COVID-19 infection and study the development of hemolytic anemia. Our main task is to analyze the results and assess subsequent changes in patients. Materials and Methods: The study was conducted among patients treated with a diagnosis of COVID-19 in the Department of Infectious Diseases at the TTA 1-Multiprofile Clinic from March to August 2023. Out of the 32 patients included, 16 were female, and 16 were male. Monitoring Blood Coagulation in Patients: The hemoglobin level of patients upon admission was initially measured using the URITEST-150 analyzer. The average for women was 110 g/l, and for men was 120 g/l. Over the course of 3 months, a decrease was observed: an average of 72 g/l in women (a decrease of up to 35%) and 84 g/l in men (a decrease of up to 30%). In the next 2 months, the positive dynamics of hemoglobin levels were observed, with an average increase to 93 g/l in women (>28%) and 112 g/l in men (>25%). Research Results: Hemolytic anemia developed in men within 5 months, reaching up to 112 g/l. In women, this process required a longer period, with the last month of observation (6 months) showing that women reached levels of up to 112 g/l, similar to men. Conclusion: Hemolytic anemia observed in patients after COVID-19 infection was monitored for 6 months (5 months in men, 6 months in women), reaching up to 112 g/l. The first 3 months after contracting COVID showed the period of development of anemia, and the subsequent 3 months indicated a stabilization period in patients.

Keywords: COVID, anemia, hemoglobin, tma, virus, viral infrection

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223 Nutritional Status of Rural Women in Bengaluru Rural District of Karnataka, India

Authors: A. M. Maruthesh, B. M. Anandakumar, O. Kumara, Akshatha Gombi, S. R. Rajini

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Women play a vital role in ensuring proper development and growth of children. They also contribute significantly towards income generation, food preparation and health. Nutritional status reflects the health of a person and is influenced by the quality of foods eaten and the ability of the body to utilize these foods to meet its needs it is affected by various socio-economic factors including income, family size, occupation and educational status of the people. The study was undertaken on nutritional status of rural women in Heggadehalli of Doddaballapurtaluk and Venkathalli of Devanahallitaluk in Bengaluru rural district with the sample size of 200 respondents. The prevalence of symptoms of malnutrition in a community is in turn a reflection of dietary consumption of its members. Mean anthropometric measurement of rural women were 153.8 cm of height, 46.8 kg of weight. In comparison with the mean BMI standards, it was observed that 20 percent of women were under nourished, 64 percent of women were normal and 16 percent women were obese. In comparison with the mean waist/hip ratio with standards, it was observed that 84 percent were in normal category and 16 percent were obese. Education, land holding, income and age had significant positive association with anthropometric measurements of rural women. The deficient level of haemoglobin existed in 53 percent of rural women, low in 20 percent and only 27 percent had acceptable level. The occurrence of morbidity symptoms was higher in rural women, its illness reported among women in the study were pain in hands and legs, backache, headache, pain in abdomen, fever, weakness, cold and cough and acidity. This may be due to considerable amount of workload on women who spend 8 to 9 hours at work and after returning continue their day’s work at home also.

Keywords: anthrometry, body index, hemoglobin, nutrient deficiency, rural women, nutritional status

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222 The Prevalence of Blood-Borne Viral Infections among Autopsy Cases in Jordan

Authors: Emad Al-Abdallat, Faris G. Bakri, Azmi Mahafza, Rayyan Al Ali, Nidaa Ababneh, Ahmed Idhair

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Background: Morgues are high-risk areas for the spread of infection from the cadavers to the staff during the postmortem examination. Infection can spread from corpses to workers by the airborne route, by direct contact, or from needle and sharp object injuries. Objective: Knowledge about the prevalence of these infections among autopsies is prudent to appreciate any risk of transmission and to further enforce safety measures. Method: A total of 242 autopsies were tested. Age ranged from 3 days to 94 years (median 75.5 years, mean 45.3 (21.9 ± SD)). There were 172 (71%) males. Results: The cause of death was considered natural in 137 (56.6%) cases, accidental in 89 (36.8%), homicidal in 9 (3.7%), suicidal in 4 (1.7%), and unknown in 3 (1.2%). Hepatitis B surface antigen was positive in 5 (2.1%) cases. Hepatitis C virus antibody was detected in 5 (2.1%) cases and the hepatitis C virus polymerase chain reaction was positive in 2 of them (0.8%). HIV antibody was not detected in any of the cases. Conclusions: Autopsies can be associated with exposure to blood borne viruses. Autopsies performed during the study period were tested for hepatitis B surface antigen, hepatitis C virus antibody, and human immunodeficiency virus antibody. Positive tests were subsequently confirmed by polymerase chain reaction. There is low prevalence of infections with these viruses in our autopsy cases. However, the risk of transmission remains a threat. Healthcare workers in the forensic departments should adhere to standard precautions.

Keywords: autopsy, hepatitis B virus, hepatitis C virus, human immunodeficiency virus, Jordan

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221 Metastatic Esophageal Squamous Cell Carcinoma Presenting with COVID-19 Infection and Cardiac Tamponade

Authors: Sutinon Yuchomsuk, Satchachon Changthom, Pruet Areesawangvong, Monsiri Jinapen

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Background: Esophageal squamous cell carcinoma can be presented with many symptoms, such as dysphagia or weight loss. However, in some circumstances, rare presentations can be found, e.g., dyspnea, which is more common in pulmonary malignancy. And dyspnea is also one of the most common presentations of COVID-19 infection. So, in this case, we can learn from many points in patient symptoms and findings leading to the diagnosis of esophageal squamous cell carcinoma. Method: This research is a case-report study including one patient from Mahasarakham Hospital, Thailand. Data were collected during December 2021. Result: A 55-year-old Thai male patient with an unknown past medical history presented with dyspnea and shortness of breath for the duration of three days prior to admission. His symptom also included cough, fever, and sore throat. Laboratory results indicated that the patient had COVID-19 pneumonia. Further investigation showed that he had cardiac tamponade and suspected pulmonary/esophageal cancer. Lung biopsy and pericardiocentesis were done, which were positive for carcinoma from pericardial effusion but negative for malignancy from the lung biopsy. Later esophagogastroduodenoscopy was done with endoscopic tissue biopsy; the result was positive for squamous cell carcinoma of the esophagus. Conclusion: Most commonly, esophageal cancer is presented with dysphagia or weight loss. However, in some rare cases, patients can also be presented with dyspnea due to cardiac tamponade. And in recent years, COVID-19 has become a pandemic all over the world, sometimes masking symptoms of other diseases. Such as in this case, the patient didn’t improve after the pneumonia was resolved, which led to the final diagnosis of metastatic esophageal cancer.

Keywords: esophageal cancer, cardiac tamponade, metastatic squamous cell carcinoma, COVID-19 infection

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220 The Association between ABO-Rh Blood Groups and the Risk of COVID-19 Infection

Authors: Abbas Sedighinejad, Hossein Khoshrang, Mohammad Haghighi, Ali Ashraf, Mostafa Saeedinia, Gelareh Biazar, Zahra Atrkarroushan, Mahdi Ajdadi

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Background: Coronavirus disease 2019 (COVID-19) as an alarming attack broke out in China and spread rapidly worldwide. Objectives: Based on a theory indicating the correlation between some viral diseases and blood types, we investigated the relationship between blood groups and coronavirus infection risk in Guilan Province, Iran. Methods: Retrospectively, all the files of the admitted patients with suspected COVID-19, in peak conditions of the disease between March 1 and May 30, 2020, were reviewed using the Census method. The required data, including epidemiological and clinical information and outcomes, were obtained from electronic records. Results: A total of 249 cases were analyzed, of whom 109 were collected from governmental centers, and the rest were collected from private hospitals. The most common co-morbidity was diabetes with 71 (37.6%) cases and the main symptom at the admission time was dyspnea with 170 (24.5%) cases. Of the total patients, 155 (62.2%) were discharged, and the rest died. The most common blood group among our patients was O Rh-positive with 91 (36.5%) cases. No meaningful correlation was found between outcomes and blood groups in terms of ABO types (P = 0.89) or Rh factor (P = 0.456). The Rh-positive proportion was significantly higher in the COVID-19 cases than in the general population (P = 0.038). Conclusions: We found that the Rh-positive proportion was significantly higher in the COVID-19 cases than in the healthy individuals. However, no correlation was observed between the groups regarding ABO typing.

Keywords: ABO blood group, COVID 19, association, personal protection

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219 Toxicity, Analgesic, and Anti-Pyretic Activities of Methanolic Extract from Hyoscyamus albus’ Leaves in Albinos Rats

Authors: Yahia Massinissa, Afaf Benhouda, Mouloud Yahia

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Objective: The aim of this study was to investigate the toxicity; analgesic and anti-pyretic properties of standardized HA methanolic extract (HAMeOH) in vivo. Methods: The acute toxicity study was performed on rats while adopting the OECD-420 Guidelines (fixed dose procedure). Assessment of analgesic activity was performed in rats with two analgesic models. One was acetic acid induced writhing response and the other formalin-induced paw licking. The anti-pyretic effect was tested by Brewer’s yeast induced fever in rats. Results: For the acute toxicity test, the higher dose administration of 2000 mg/kg bw. of H.albus did not produce any toxic signs or deaths in rats. There were no significant differences (p>0.05) in the body and organ weights between control and treated groups. The (LD50) of 'H. albus' was higher than 2000 g/kg bw. In subacute toxicity study, no mortality and toxic signs were observed with the doses of 100 and 200 mg/kg bw. of extracts of for 28 consecutive days. These analgesic experimental results indicated that HAMeOH (100 mg/kg and 200 mg/kg) decreased the acetic acid-induced writhing responses and HAMeOH (100 mg/kg and 200 mg/kg) decreased the licking time in the second phase of the formalin test. Moreover, in the model of yeast-induced elevation of the body temperature HAMeOH showed dose-dependent lowering of the body temperature up to 3h at both the doses these results obtained, were comparable to that of paracetamol. Conclusion: The present findings indicate that the leaves of Hyoscyamus albus L. possess potent analgesic and antipyretic activity.

Keywords: Hyoscyamus albus, Umbilicus rupestris, secondary metabolites, NMR with protons, pharmacobiologic activities, methanolic extract

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218 Evaluation of Immunostimulant Potential of Proteoliposomes Derived from Vibrio anguillarum Administered by Immersion in Zebrafish (Danio rerio)

Authors: M. Caruffo, P. Navarrete, C. G. Feijoo, L. Sáenz

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Disease prevention through the use of vaccines has been crucial to achieve the current level of production in the salmon industry. However, vaccines have been developed based largely on inactivated bacterial formulations, using the whole pathogen. These formulations have demonstrated excellent efficacy against extracellular bacterial pathogens. However diseases with the greatest economic impacts correspond to intracellular bacterial and viral pathogens, vaccines based on these types of agents have shown a discrete effectiveness. It is for these reasons that the development of subunit vaccines based on defined antigens offers a promising solution. The main problem is that subunit vaccines offer a low immunogenicity, since they lack immunostimulatory elements, so that the development of new adjuvants platforms becomes an important challenge for this type of formulations. We evaluate the effect of a formulation based on proteoliposomes of Vibrio anguillarum administered by immersion as a new adjuvant strategy, allowing efficient stimulation of the innate immune system. Proteoliposomes physicochemical properties were evaluated in its ability to produce an inflammatory process. Using zebrafish (Danio rerio) larvae as a model species and the transgenic line (Tg(mpx: GFP)i114) allowed us to track the neutrophil migration in real time. Additionally we evaluated the gene expression of some molecular markers involved in the development of the innate immune response characterizing the adjuvant capacity of the formulation.

Keywords: adjuvants, vaccine development, zebrafish, innate immunity

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217 The Investigation of Effect of Alpha Lipoic Acid against Damage on Neonatal Rat Lung to Maternal Tobacco Smoke Exposure

Authors: Elif Erdem, Nalan Kaya, Gonca Ozan, Durrin Ozlem Dabak, Enver Ozan

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This study was carried out to determine the histological and biochemical changes in the lungs of the rat pups exposed to tobacco smoke during pregnancy period and to investigate the protective effects of alpha lipoic acid, which is administered during pregnancy, on these changes. In our study, 24 six-week old Spraque-Dawley female rats weighing 160 ± 10 g were used (n:7). Rats were randomly divided into four equal groups: group I (control), group II (tobacco smoke), group III (tobacco smoke + alpha lipoic acid) and group IV (alpha lipoic acid). Rats in the group II, group III were exposed to tobacco smoke twice a day for one hour starting from eight weeks before mating and during pregnancy. In addition to tobacco smoke, 20 mg/kg of alpha lipoic acid was administered via oral gavage to the rats in the group III. Only alpha lipoic acid was administered to the rats in the group IV. Once after the delivery, all administrations were stopped. On the 7 and 21th days, the seven pups of all groups were decapitated. A portion of the lung was taken and stained with HE, PAS and Masson. In addition to immunohistochemical staining of surfactant protein A, vascular endothelial growth factor, caspase-3, TUNEL method was also used to determine apoptosis. Biochemical analyzes were performed with some part of the lung tissue specimens. In the histological evaluations performed under light microscopy, inflammatory cell increase, hemorrhagic areas, edema, interalveolar septal thickening, alveolar numbers decrease, degeneration of some bronchi and bronchial epithelium, epithelial cells that were fallen into the lumen and hyaline membrane formation were observed in tobacco smoke group. These findings were ameliorated in tobacco smoke + ALA group. Hyaline membrane formation was not detected in this group. The TUNEL positive cell numbers a significant increase was detected in the tobacco smoke group, whereas a significant decrease was detected in the tobacco smoke + ALA group. In terms of the immunoreactivity of both SP-A and VEGF, a significant decrease was observed in the tobacco smoke group, and a significant increase was observed in the tobacco smoke + ALA group. Regarding the immunoreactivity of caspase-3, there was a significant increase in the group of tobacco smoke and a significant decrease in the group of tobacco smoke + ALA. The malondialdehyde levels were determined to be significantly increased in the tobacco smoke group, and a significant decreased in the tobacco smoke + ALA. Glutathione and superoxide dismutase enzyme activities showed a significant decrease in the group of tobacco smoke and a significant increase in the tobacco smoke + ALA group. In conclusion, we suggest that the exposure to tobacco smoke during pregnancy leads to morphological, histopathological and functional changes on lung development by causing oxidative damage in lung tissues of neonatal rats and the maternal use of alpha lipoic acid can provide a protective effect on the neonatal lung development against this oxidative stress originating from tobacco smoke.

Keywords: alpha lipoic acid, lung, neonate, tobacco smoke, pregnancy

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216 Preparation of Pegylated Interferon Alpha-2b with High Antiviral Activity Using Linear 20 KDa Polyethylene Glycol Derivative

Authors: Ehab El-Dabaa, Omnia Ali, Mohamed Abd El-Hady, Ahmed Osman

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Recombinant human interferon alpha 2 (rhIFN-α2) is FDA approved for treatment of some viral and malignant diseases. Approved pegylated rhIFN-α2 drugs have highly improved pharmacokinetics, pharmacodynamics and therapeutic efficiency compared to native protein. In this work, we studied the pegylation of purified properly refolded rhIFN-α2b using linear 20kDa PEG-NHS (polyethylene glycol- N-hydroxysuccinimidyl ester) to prepare pegylated rhIFN-α2b with high stability and activity. The effect of different parameters like rhIFN-α2b final concentration, pH, rhIFN-α2b/PEG molar ratios and reaction time on the efficiency of pegylation (high percentage of monopegylated rhIFN-α2b) have been studied in small scale (100µl) pegylation reaction trials. Study of the percentages of different components of these reactions (mono, di, polypegylated rhIFN-α2b and unpegylated rhIFN-α2b) indicated that 2h is optimum time to complete the reaction. The pegylation efficiency increased at pH 8 (57.9%) by reducing the protein concentration to 1mg/ml and reducing the rhIFN-α2b/PEG ratio to 1:2. Using larger scale pegylation reaction (65% pegylation efficiency), ion exchange chromatography method has been optimized to prepare and purify the monopegylated rhIFN-α2b with high purity (96%). The prepared monopegylated rhIFN-α2b had apparent Mwt of approximately 65 kDa and high in vitro antiviral activity (2.1x10⁷ ± 0.8 x10⁷ IU/mg). Although it retained approximately 8.4 % of the antiviral activity of the unpegylated rhIFN-α2b, its activity is high compared to other pegylated rhIFN-α2 developed by using similar approach or higher molecular weight branched PEG.

Keywords: antiviral activity, rhIFN-α2b, pegylation, pegylation efficiency

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