Search results for: Phage vaccine
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 260

Search results for: Phage vaccine

170 Using Baculovirus Expression Vector System to Express Envelop Proteins of Chikungunya Virus in Insect Cells and Mammalian Cells

Authors: Tania Tzong, Chao-Yi Teng, Tzong-Yuan Wu

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Currently, Chikungunya virus (CHIKV) transmitted to humans by Aedes mosquitoes has distributed from Africa to Southeast Asia, South America, and South Europe. However, little is known about the antigenic targets for immunity, and there are no licensed vaccines or specific antiviral treatments for the disease caused by CHIKV. Baculovirus has been recognized as a novel vaccine vector with attractive characteristic features of an optional vaccine delivery vehicle. This approach provides the safety and efficacy of CHIKV vaccine. In this study, bi-cistronic recombinant baculoviruses vAc-CMV-CHIKV26S-Rhir-EGFP and vAc-CMV-pH-CHIKV26S-Lir-EGFP were produced. Both recombinant baculovirus can express EGFP reporter gene in insect cells to facilitate the recombinant virus isolation and purification. Examination of vAc-CMV-CHIKV26S-Rhir-EGFP and vAc-CMV-pH-CHIKV26S-Lir-EGFP showed that this recombinant baculovirus could induce syncytium formation in insect cells. Unexpectedly, the immunofluorescence assay revealed the expression of E1 and E2 of CHIKV structural proteins in insect cells infected by vAc-CMV-CHIKV26S-Rhir-EGFP. This result may imply that the CMV promoter can induce the transcription of CHIKV26S in insect cells. There are also E1 and E2 expression in mammalian cells transduced by vAc-CMV-CHIKV26S-Rhir-EGFP and vAc-CMV-pH-CHIKV26S-Lir-EGFP. The expression of E1 and E2 proteins of insect and mammalian cells was validated again by Western blot analysis. The vector construction with dual tandem promoters, which is polyhedrin and CMV promoter, has higher expression of the E1 and E2 of CHIKV structural proteins than the vector construction with CMV promoter only. Most of the E1 and E2 proteins expressed in mammalian cells were glycosylated. In the future, the expression of structural proteins of CHIKV in mammalian cells is expected can form virus-like particle, so it could be used as a vaccine for chikungunya virus.

Keywords: chikungunya virus, virus-like particle, vaccines, baculovirus expression vector system

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169 Improving Infant Vaccination Rates Through Expanded Access to Care

Authors: Aidan Jacobsen, Morgan Motia, David Sam, Jonathan Mudge

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Background: The Centers for Disease Control (CDC) lists vaccine requirements for children under two years old to correlate with development markers. CDC lists the coverage by age 24 months to be at least 90% nationally and 84% for Rhode Island Blackstone Valley Community Health Center (BVCHC) in Central Falls, Rhode Island, currently has a completed vaccination rate of 51% for children by the age of 24 months. Current barriers to care for up to date well child vaccinations include lack of transportation, parental work, childcare, and other social stressors. Objective: Increase the vaccination rate of children under the age of 24 months at BVCHC. Conduct a literature review to identify the common barriers preventing children under 24 months from receiving vaccinations. Reduce the barriers to expand access to vaccination care for infants Methods: Setting: Blackstone Valley Community Health Center, Pawtucket, RI Participants: (n=41), Patients between the age of 20-24 months, not up to date with the CDC vaccination recommendations and without a future appointment. QI Intervention: Patients were contacted via phone and offered an appointment during extra Saturday clinic hours in order to receive up to date vaccine care. A Saturday vaccine clinic was established specifically for patients in need of vaccines and having identified barriers to care. Conclusions: Expanding clinic hours and targeting non vaccine up –to-date patients can increase the current standard of childhood immunizations at BVCHC. Overcoming barriers preventing childhood immunization can improve access to providing up to date vaccinations. Other barriers still deter from reaching the national standard of immunizations rates.

Keywords: vaccinations, well child care, barriers to care, social determinants of health

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168 Isolation and Characterization of Actinophages Infecting Streptomyces scabies in Egypt

Authors: D. Zahran, M. AlKhazindar, M. Khalil, E. T. A. Sayed

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Streptomyces scabies is a pathogenic actinomycete that infects potato crop causing severe production losses. Actinophages affect the composition and diversity of the bacterial population, thereby, can be used as a biological control. Samples of actinomycetes and phages were collected from different cultivated soils including farms of Faculty of Science, Faculty of Agriculture and different locations in Giza, Egypt. Actinomycetes were identified by using biochemical, morphological tests and molecular studies using 16S rRNA sequencing. Two specific phages (E1 and E2) against Streptomyces scabies and other hosts were isolated. Phages were identified using dilution end point (DEP), longevity in vitro (LIV), thermal inactivation point (TIP), host range and electron microscopy. PhageE1 was characterized by 10-8 (DEP),180 days(LIV), 95°C(TIP), narrow host range and electron microscopy showed ahead (59.9 nm) and neck (10.4nm). On the other hand phageE2 had 10-20 (DEP),180 days(LIV), 90°C(TIP), and the size of head was (67.2 nm) and tail (114nm). Antiviral activity was also studied using different chemicals (NaCL, KCL, CaCL2, BaCL2, CoCL2, AgNO3, ALCL3and HgCL2) with different concentrations and different plant extracts with different concentrations (star anise, tea, tillia, peppermint, ginger, cumin, chamomile, turmeric cinnamon, marjoram and black cumin). Both Phage E1and phage E2 were vulnerable to (cumin, ginger, chamomile, guavas leaves and star anise) but resistant to (Tillie, marjoram, fennelflower seeds, peppermint, and cinnamon).

Keywords: potato scab, actinophages, biological control, electron microscopy, TIP, DEP, LIV, antiviral activity

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167 A Novel Epitope Prediction for Vaccine Designing against Ebola Viral Envelope Proteins

Authors: Manju Kanu, Subrata Sinha, Surabhi Johari

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Viral proteins of Ebola viruses belong to one of the best studied viruses; however no effective prevention against EBOV has been developed. Epitope-based vaccines provide a new strategy for prophylactic and therapeutic application of pathogen-specific immunity. A critical requirement of this strategy is the identification and selection of T-cell epitopes that act as vaccine targets. This study describes current methodologies for the selection process, with Ebola virus as a model system. Hence great challenge in the field of ebola virus research is to design universal vaccine. A combination of publicly available bioinformatics algorithms and computational tools are used to screen and select antigen sequences as potential T-cell epitopes of supertypes Human Leukocyte Antigen (HLA) alleles. MUSCLE and MOTIF tools were used to find out most conserved peptide sequences of viral proteins. Immunoinformatics tools were used for prediction of immunogenic peptides of viral proteins in zaire strains of Ebola virus. Putative epitopes for viral proteins (VP) were predicted from conserved peptide sequences of VP. Three tools NetCTL 1.2, BIMAS and Syfpeithi were used to predict the Class I putative epitopes while three tools, ProPred, IEDB-SMM-align and NetMHCII 2.2 were used to predict the Class II putative epitopes. B cell epitopes were predicted by BCPREDS 1.0. Immunogenic peptides were identified and selected manually by putative epitopes predicted from online tools individually for both MHC classes. Finally sequences of predicted peptides for both MHC classes were looked for common region which was selected as common immunogenic peptide. The immunogenic peptides were found for viral proteins of Ebola virus: epitopes FLESGAVKY, SSLAKHGEY. These predicted peptides could be promising candidates to be used as target for vaccine design.

Keywords: epitope, b cell, immunogenicity, ebola

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166 Chitosan Modified Halloysite Nanomaterials for Efficient and Effective Vaccine Delivery in Farmed Fish

Authors: Saji George, Eng Khuan Seng, Christof Luda

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Nanotechnology has been recognized as an important tool for modern agriculture and has the potential to overcome some of the pressing challenges faced by aquaculture industry. A strategy for optimizing nanotechnology-based therapeutic delivery platform for immunizing farmed fish was developed. Accordingly, a compositional library of nanomaterials of natural chemistry (Halloysite (clay), Chitosan, Hydroxyapatite, Mesoporous Silica and a composite material of clay-chitosan) was screened for their toxicity and efficiency in delivering models antigens in cellular and zebrafish embryo models using high throughput screening platforms. Through multi-parametric optimization, chitosan modified halloysite (clay) nanomaterial was identified as an optimal vaccine delivery platform. Further, studies conducted in juvenile seabass showed the potential of clay-chitosan in delivering outer membrane protein of Tenacibaculum maritimum- TIMA (pathogenic bacteria) to and its efficiency in eliciting immune responses in fish. In short, as exemplified by this work, the strategy of using compositional nanomaterial libraries and their biological profiling using high-throughput screening platform could fasten the discovery process of nanomaterials with potential applications in food and agriculture.

Keywords: nanotechnology, fish-vaccine, drug-delivery, halloysite-chitosan

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165 Expression of Human Papillomavirus Type 18 L1 Virus-Like Particles in Methylotropic Yeast, Pichia Pastoris

Authors: Hossein Rassi, Marjan Moradi Fard, Samaneh Niko

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Human papillomavirus type 16 and 18 are closely associated with the development of human cervical carcinoma, which is one of the most common causes of cancer death in women worldwide. At present, HPV type 18 accounts for about 34 % of all HPV infections in Iran and the most promising vaccine against HPV infection is based on the L1 major capsid protein. The L1 protein of HPV18 has the capacity to self-assemble into capsomers or virus-like particles (VLPs) that are non-infectious, highly immunogenic and allowing their use in vaccine production. The methylotrophic yeast Pichia pastoris is an efficient and inexpensive expression system used to produce high levels of heterologous proteins. In this study we expressed HPV18 L1 VLPs in P. pastoris. The gene encoding the major capsid protein L1 of the high-risk HPV type 18 was isolated from Iranian patient by PCR and inserted into pTG19-T vector to obtain the recombinant expression vector pTG19-HPV18-L1. Then, the pTG19-HPV18-L1 was transformed into E. coli strain DH5α and the recombinant protein HPV18 L1 was expressed under IPTG induction in soluble form. The HPV18 L1 gene was excised from recombinant plasmid with XhoI and EcoRI enzymes and ligated into the yeast expression vector pPICZα linearized with the same enzymes, and transformed into P. pastoris. Induction and expression of HPV18 L1 protein was demonstrated by BMGY/BMMY and RT PCR. The parameters for induced cultivation for strain in P. pastoris KM71 with HPV16L1 were investigated in shaking flask cultures. After induced cultivation BMMY (pH 7.0) medium supplemented with methanol to a final concentration of 1.0% every 24 h at 37 degrees C for 96 h, the recombinant produced 78.6 mg/L of L1 protein. This work offers the possibility for the production of prophylactic vaccine for cervical carcinoma by P. pastoris for HPV-18 L1 gene. The VLP-based HPV vaccines can prevent persistent HPV18 infections and cervical cancer in Iran. The HPV-18 L1 gene was expressed successfully in E.coli, which provides necessary basis for preparing HPV-18 L1 vaccine in human. Also, HPV type 6 L1 proteins expressed in Pichia pastoris will facilitate the HPV vaccine development and structure-function study.

Keywords: Pichia pastoris, L1 virus-like particles, human papillomavirus type 18, biotechnology

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164 Expectations of Unvaccinated Health Workers in Greece and the Question of Trust: A Qualitative Study of Vaccine Hesitancy

Authors: Sideri Katerina, Chanania Eleni

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The reasons why people remain unvaccinated, especially health workers, are complex. In Greece, 2 percent of health workers (around 7,000) remain unvaccinated, despite the fact that for this group of people vaccination against COVID-19 is mandatory. In April 2022, the Greek health minister repeated that unvaccinated health care workers will remain suspended from their jobs ‘for as long as the pandemic lasts,’ explaining that the suspension of the workers in question was ‘entirely their choice’ and that health professionals who do not believe in vaccines ‘do not believe in their own science.’ Although policy circles around the world often link vaccine hesitancy to ignorance of science or misinformation, various recently published qualitative studies show that vaccine hesitancy is the result of a combination of factors, which include distrust towards elites and the system of innovation and distrust towards government. In a similar spirit, some commentators warn that labeling hesitancy as “anti-science” is bad politics. In this paper, we worked within the tradition of STS taking the view that people draw upon personal associations to enact and express civic concern with an issue, the enactment of public concern involves the articulation of threats to actors’ way of life, personal values, relationships, lived experiences, broader societal values and institutional structures. To this effect, we have conducted 27 in depth interviews with unvaccinated Greek health workers and we are in the process of conducting 20 more interviews. We have so far found that rather than a question of believing in ‘facts’ vaccine hesitancy reflects deep distrust towards those charged with the making of decisions and pharmaceutical companies and that emotions (rather than rational thinking) play a crucial role in the formation of attitudes and the making of decisions. We need to dig deeper so as to understand the causes of distrust towards technical government and the ways in which public(s) conceive of and want to be part in the politics of innovation. We particularly address the question of the effectiveness of mandatory vaccination of health workers and whether such top-down regulatory measures further polarize society, to finally discuss alternative regulatory approaches and governance structures.

Keywords: vaccine hesitancy, innovation, trust in vaccines, sociology of vaccines, attitude drivers towards scientific information, governance

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163 Molecular Farming: Plants Producing Vaccine and Diagnostic Reagent

Authors: Katerina H. Takova, Ivan N. Minkov, Gergana G. Zahmanova

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Molecular farming is the production of recombinant proteins in plants with the aim to use the protein as a purified product, crude extract or directly in the planta. Plants gain more attention as expression systems compared to other ones due to the cost effective production of pharmaceutically important proteins, appropriate post-translational modifications, assembly of complex proteins, absence of human pathogens to name a few. In addition, transient expression in plant leaves enables production of recombinant proteins within few weeks. Hepatitis E virus (HEV) is a causative agent of acute hepatitis. HEV causes epidemics in developing countries and is primarily transmitted through the fecal-oral route. Presently, all efforts for development of Hepatitis E vaccine are focused on the Open Read Frame 2 (ORF2) capsid protein as it contains epitopes that can induce neutralizing antibodies. For our purpose, we used the CMPV-based vector-pEAQ-HT for transient expression of HEV ORF2 in Nicotiana benthamina. Different molecular analysis (Western blot and ELISA) showed that HEV ORF2 capsid protein was expressed in plant tissue in high-yield up to 1g/kg of fresh leaf tissue. Electron microscopy showed that the capsid protein spontaneously assembled in low abundance virus-like particles (VLPs), which are highly immunogenic structures and suitable for vaccine development. The expressed protein was recognized by both human and swine HEV positive sera and can be used as a diagnostic reagent for the detection of HEV infection. Production of HEV capsid protein in plants is a promising technology for further HEV vaccine investigations. Here, we reported for a rapid high-yield transient expression of a recombinant protein in plants suitable for vaccine production as well as a diagnostic reagent. Acknowledgments -The authors’ research on HEV is supported with grants from the Project PlantaSYST under the Widening Program, H2020 as well as under the UK Biotechnological and Biological Sciences Research Council (BBSRC) Institute Strategic Programme Grant ‘Understanding and Exploiting Plant and Microbial Secondary Metabolism’ (BB/J004596/1). The authors want to thank Prof. George Lomonossoff (JIC, Norwich, UK) for his contribution.

Keywords: hepatitis E virus, plant molecular farming, transient expression, vaccines

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162 Production of Recombinant VP2 Protein of Canine Parvovirus Type 2c Using Baculovirus Expression System

Authors: Jae Young Song, In-Ohk Ouh, Seyeon Park, Byeong Sul Kang, Soo Dong Cho, In-Soo Cho

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Canine parvovirus (CPV) is a major pathogen of diarrhea disease in dogs. CPV type 2 has three of antigenic variants such as 2a, 2b, and 2c. CPV constructs a small non-enveloped, icosahedral capsid that contains single-stranded DNA. It has capsids that two largely overlapping virion proteins (VP), VP1 (82 kDa), and VP2 (65 kDa). Baculoviruses are insect pathogens that regulate insect populations in nature and are being successfully used to control insect pests. The proteins produced in the baculovirus-expression system are used for instance for functional studies, vaccine preparations, or diagnostics. The vaccines produced by baculovirus-expression system showed elicitation of antibodies. The recombinant baculovirus infected SF9 cells showed broken shape. The recombinant VP2 proteins from cell pellet or supernatant were confirmed by western blotting. The result showed that the recombinant VP2 protein bands were appeared at 65 kDa molecular weight in both cell pellet and supernatant of infected SF9 cell. These results indicated that the recombinant baculovirus infected SF9 cell express the recombinant VP2 protein successfully. In addition, the expressed recombinant VP2 protein is secreted from cell to supernatant. The baculovirus expression system can be used to produce the VP2 protein of CPV 2c. In addition, the secretion property of the expression of VP2 protein may decrease the cost of production, because it can be skipped the cell breaking step. The produced VP2 protein could be used for vaccine and the agent of diagnostic tests. This study provides the foundation of the production of CPV 2c vaccine and the diagnostic agent.

Keywords: baculovirus, canine parvovirus 2c, dog, Korea

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161 Safety Profile of Human Papillomavirus Vaccines: A Post-Licensure Analysis of the Vaccine Adverse Events Reporting System, 2007-2017

Authors: Giulia Bonaldo, Alberto Vaccheri, Ottavio D'Annibali, Domenico Motola

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The Human Papilloma Virus (HPV) was shown to be the cause of different types of carcinomas, first of all of the cervical intraepithelial neoplasia. Since the early 80s to today, thanks first to the preventive screening campaigns (pap-test) and following to the introduction of HPV vaccines on the market; the number of new cases of cervical cancer has decreased significantly. The HPV vaccines currently approved are three: Cervarix® (HPV2 - virus type: 16 and 18), Gardasil® (HPV4 - 6, 11, 16, 18) and Gardasil 9® (HPV9 - 6, 11, 16, 18, 31, 33, 45, 52, 58), which all protect against the two high-risk HPVs (6, 11) that are mainly involved in cervical cancers. Despite the remarkable effectiveness of these vaccines has been demonstrated, in the recent years, there have been many complaints about their risk-benefit profile due to Adverse Events Following Immunization (AEFI). The purpose of this study is to provide a support about the ongoing discussion on the safety profile of HPV vaccines based on real life data deriving from spontaneous reports of suspected AEFIs collected in the Vaccine Adverse Events Reporting System (VAERS). VAERS is a freely-available national vaccine safety surveillance database of AEFI, co-administered by the Centers for Disease Control and Prevention (CDC) and Food and Drug Administration (FDA). We collected all the reports between January 2007 to December 2017 related to the HPV vaccines with a brand name (HPV2, HPV4, HPV9) or without (HPVX). A disproportionality analysis using Reporting Odds Ratio (ROR) with 95% confidence interval and p value ≤ 0.05 was performed. Over the 10-year period, 54889 reports of AEFI related to HPV vaccines reported in VAERS, corresponding to 224863 vaccine-event pairs, were retrieved. The highest number of reports was related to Gardasil (n = 42244), followed by Gardasil 9 (7212) and Cervarix (3904). The brand name of the HPV vaccine was not reported in 1529 cases. The two events more frequently reported and statistically significant for each vaccine were: dizziness (n = 5053) ROR = 1.28 (CI95% 1.24 – 1.31) and syncope (4808) ROR = 1.21 (1.17 – 1.25) for Gardasil. For Gardasil 9, injection site pain (305) ROR = 1.40 (1.25 – 1.57) and injection site erythema (297) ROR = 1.88 (1.67 – 2.10) and for Cervarix, headache (672) ROR = 1.14 (1.06 – 1.23) and loss of consciousness (528) ROR = 1.71 (1.57 – 1.87). In total, we collected 406 reports of death and 2461 cases of permanent disability in the ten-year period. The events consisting of incorrect vaccine storage or incorrect administration were not considered. The AEFI analysis showed that the most frequently reported events are non-serious and listed in the corresponding SmPCs. In addition to these, potential safety signals arose regarding less frequent and severe AEFIs that would deserve further investigation. This already happened with the referral of the European Medicines Agency (EMA) for the adverse events POTS (Postural Orthostatic Tachycardia Syndrome) and CRPS (Complex Regional Pain Syndrome) associated with anti-papillomavirus vaccines.

Keywords: adverse drug reactions, pharmacovigilance, safety, vaccines

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160 Lessons Learned from Push-Plus Implementation in Northern Nigeria

Authors: Aisha Giwa, Mohammed-Faosy Adeniran, Olufunke Femi-Ojo

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Four decades ago, the World Health Organization (WHO) launched the Expanded Programme on Immunization (EPI). The EPI blueprint laid out the technical and managerial functions necessary to routinely vaccinate children with a limited number of vaccines, providing protection against diphtheria, tetanus, whooping cough, measles, polio, and tuberculosis, and to prevent maternal and neonatal tetanus by vaccinating women of childbearing age with tetanus toxoid. Despite global efforts, the Routine Immunization (RI) coverage in two of the World Health Organization (WHO) regions; the African Region and the South-East Asia Region, still remains short of its targets. As a result, the WHO Regional Director for Africa declared 2012 as the year for intensifying RI in these regions and this also coincided with the declaration of polio as a programmatic emergency by the WHO Executive Board. In order to intensify routine immunization, the National Routine Immunization Strategic Plan (2013-2015) stated that its core priority is to ensure 100% adequacy and availability of vaccines for safe immunization. To achieve 100% availability, the “PUSH System” and then “Push-Plus” were adopted for vaccine distribution, which replaced the inefficient “PULL” method. The NPHCDA plays the key role in coordinating activities in area advocacy, capacity building, engagement of 3PL for the state as well as monitoring and evaluation of the vaccine delivery process. eHealth Africa (eHA) is a player as a 3PL service provider engaged by State Primary Health Care Boards (SPHCDB) to ensure vaccine availability through Vaccine Direct Delivery (VDD) project which is essential to successful routine immunization services. The VDD project ensures the availability and adequate supply of high-quality vaccines and immunization-related materials to last-mile facilities. eHA’s commitment to the VDD project saw the need for an assessment of the project vis-a-vis the overall project performance, evaluation of a process for necessary improvement suggestions as well as general impact across Kano State (Where eHA had transitioned to the state), Bauchi State (currently manage delivery to all LGAs except 3 LGAs currently being managed by the state), Sokoto State (eHA currently covers all LGAs) and Zamfara State (Currently, in-sourced and managed solely by the state).

Keywords: cold chain logistics, health supply chain system strengthening, logistics management information system, vaccine delivery traceability and accountability

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159 Health Communication: A Southwest Georgia Health Literacy Project

Authors: Marsha R. Lawrence

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Introduction: In February and March of 2020, many Black Americans in Albany, Georgia, were impacted by COVID-19 compared to the rest of the country. Due to misinformation and distrust in the community, citizens were not able to make good health decisions regarding COVID-19. The city of Albany applied for a grant with the Department of Health and Human Services, specifically the Office of Minority Health and it was approved. The city of Albany partnered with Albany State University to administer the grant and implementation ensued. Method: An eleven-page electronic and paper cross-sectional survey was given to participants. Albany State University recruited community partners like health care organizations and faith-based organizations to reach the citizens of Albany, Georgia. These partners reached participants through creative community activities to educate participants about COVID-19 and provide incentives to receive a vaccine. Data collection is still in progress because activities are ongoing. Anticipated Results: By December 2023, we anticipate results of the number of participants who accepted vaccines based on participants who stated providers checked their understanding, participants who were satisfied with communication regarding COVID-19 health information about the vaccine, and participants who were involved in decisions regarding the COVID-19 vaccine. Conclusion: Health communication is a subsection of health literacy. At this point, approximately 4000 individuals have received information and education about COVID-19 in the Albany area. We expect building trusting relationships played an important part in the increase in knowledge and vaccination in Albany, Georgia.

Keywords: health literacy, health communication, vaccination, COVID-19

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158 A Statistical Analysis on the Comparison of First and Second Waves of COVID-19 and Importance of Early Actions in Public Health for Third Wave in India

Authors: Maitri Dave

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Coronaviruses (CoV) is such infectious virus which has impacted globally in a more dangerous manner causing severe lung problems and leaving behind more serious diseases among the people. This pandemic has affected globally and created severe respiratory problems, and damaged the lungs. India has reported the first case of COVID-19 in January 2020. The first wave of COVID-19 took place from April to September of 2020. Soon after, a second peak is also noticed in the month of March 2021, which in turn becomes more dangerous due to a lack of supply of medical equipment. It created resource deficiency globally, specifically in India, where some necessary life-saving equipment like ventilators and oxygenators were not sufficient to cater to the demand-supply ratio effectively. Through carefully examining such a situation, India began to execute the process of vaccination in the month of January 2021 and successfully administered 25,46,71,259 doses of vaccines till now, which is only 15.5% of the total population while only 3.6% of the total population is fully vaccinated. India has authorized the British Oxford–AstraZeneca vaccine (Covishield), the Indian BBV152 (Covaxin) vaccine, and the Russian Sputnik V vaccine for emergency use. In the present study, we have collected all the data state wisely of both first and second wave and analyzed them using MS Excel Version 2019 and SPSS Statistics Version 26. Following the trends, we have predicted the characteristics of the upcoming third wave of COVID-19 and recommended some strategies, early actions, and measures that can be taken by the public health system in India to combat the third wave more effectively.

Keywords: COVID-19, vaccination, Covishiled, Coronavirus

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157 Efficacy and Safety of COVID-19 Vaccination in Patients with Multiple Sclerosis: Looking Forward to Post-COVID-19

Authors: Achiron Anat, Mathilda Mandel, Mayust Sue, Achiron Reuven, Gurevich Michael

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Introduction: As coronavirus disease 2019 (COVID-19) vaccination is currently spreading around the world, it is of importance to assess the ability of multiple sclerosis (MS) patients to mount an appropriate immune response to the vaccine in the context of disease-modifying treatments (DMT’s). Objectives: Evaluate immunity generated following COVID-19 vaccination in MS patients, and assess factors contributing to protective humoral and cellular immune responses in MS patients vaccinated against severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) virus infection. Methods: Review our recent data related to (1) the safety of PfizerBNT162b2 COVID-19 mRNA vaccine in adult MS patients; (2) the humoral post-vaccination SARS-CoV2 IgG response in MS vaccinees using anti-spike protein-based serology; and (3) the cellular immune response of memory B-cells specific for SARS-CoV-2 receptor-binding domain (RBD) and memory T-cells secreting IFN-g and/or IL-2 in response to SARS-CoV2 peptides using ELISpot/Fluorospot assays in MS patients either untreated or under treatment with fingolimod, cladribine, or ocrelizumab; (4) covariate parameters related to mounting protective immune responses. Results: COVID-19 vaccine proved safe in MS patients, and the adverse event profile was mainly characterised by pain at the injection site, fatigue, and headache. Not any increased risk of relapse activity was noted and the rate of patients with acute relapse was comparable to the relapse rate in non-vaccinated patients during the corresponding follow-up period. A mild increase in the rate of adverse events was noted in younger MS patients, among patients with lower disability, and in patients treated with DMTs. Following COVID-19 vaccination protective humoral immune response was significantly decreased in fingolimod- and ocrelizumab- treated MS patients. SARS-CoV2 specific B-cell and T-cell cellular responses were respectively decreased. Untreated MS patients and patients treated with cladribine demonstrated protective humoral and cellular immune responses, similar to healthy vaccinated subjects. Conclusions: COVID-19 BNT162b2 vaccine proved as safe for MS patients. No increased risk of relapse activity was noted post-vaccination. Although COVID-19 vaccination is new, accumulated data demonstrate differences in immune responses under various DMT’s. This knowledge can help to construct appropriate COVID-19 vaccine guidelines to ensure proper immune responses for MS patients.

Keywords: covid-19, vaccination, multiple sclerosis, IgG

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156 Process Development of pVAX1/lacZ Plasmid DNA Purification Using Design of Experiment

Authors: Asavasereerat K., Teacharsripaitoon T., Tungyingyong P., Charupongrat S., Noppiboon S. Hochareon L., Kitsuban P.

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Third generation of vaccines is based on gene therapy where DNA is introduced into patients. The antigenic or therapeutic proteins encoded from transgenes DNA triggers an immune-response to counteract various diseases. Moreover, DNA vaccine offers the customization of its ability on protection and treatment with high stability. The production of DNA vaccines become of interest. According to USFDA guidance for industry, the recommended limits for impurities from host cell are lower than 1%, and the active conformation homogeneity supercoiled DNA, is more than 80%. Thus, the purification strategy using two-steps chromatography has been established and verified for its robustness. Herein, pVax1/lacZ, a pre-approved USFDA DNA vaccine backbone, was used and transformed into E. coli strain DH5α. Three purification process parameters including sample-loading flow rate, the salt concentration in washing and eluting buffer, were studied and the experiment was designed using response surface method with central composite face-centered (CCF) as a model. The designed range of selected parameters was 10% variation from the optimized set point as a safety factor. The purity in the percentage of supercoiled conformation obtained from each chromatography step, AIEX and HIC, were analyzed by HPLC. The response data were used to establish regression model and statistically analyzed followed by Monte Carlo simulation using SAS JMP. The results on the purity of the product obtained from AIEX and HIC are between 89.4 to 92.5% and 88.3 to 100.0%, respectively. Monte Carlo simulation showed that the pVAX1/lacZ purification process is robust with confidence intervals of 0.90 in range of 90.18-91.00% and 95.88-100.00%, for AIEX and HIC respectively.

Keywords: AIEX, DNA vaccine, HIC, puification, response surface method, robustness

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155 Factors Associated with Seroconversion of Oral Polio Vaccine among the Children under 5 Year in District Mirpurkhas, Pakistan 2015

Authors: Muhammad Asif Syed, Mirza Amir Baig

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Background: Pakistan is one of the two remaining polio-endemic countries, posing a significant public health challenge for global polio eradication due to failure to interrupt polio transmission. Country specific seroprevalence studies help in the evaluation of immunization program performance, the susceptibility of population against polio virus and identification of existing level of immunity with factors that affect seroconversion of the oral polio vaccine (OPV). The objective of the study was to find out factors associated with seroconversion of the OPV among children 6-59 months in Pakistan. Methods: A Hospital based cross-sectional serosurvey was undertaken in May-June 2015 at District Mirpurkhas, Sindh-Pakistan. Total 180 children aged 6–59 months were selected by using systematic random sampling from Muhammad Medical College Hospital, Mirpurkhas. Demographic, vaccination history and risk factors information were collected from the parents/guardian. Blood sample was collected and tested for the detection of poliovirus IgG antibodies by using ELISA Kit. The IgG titer <10 IU/ml, 50 to <150 IU/ml and >150 IU/ml was defined as negative, weak positive and positive immunity respectively. Pearson Chi-square test was used to determine the difference in seroprevalence in univariate analysis. Results: A total of 180 subjects were enrolled mean age was 23 months (7 -59 months). Off these 160 (89%) children were well and 18 (10%) partially protected against polio virus. Two (1.1%) children had no protection against polio virus as they had <10 IU/ml poliovirus IgG antibodies titer. Both negative cases belong from the female gender, age group 12-23 months, urban area and BMI <50 percentile. There was a difference between normal and the wasting children; it did attain statistical significance (χ2= 35.5, p=0.00). The difference in seroconversion was also observed in relation to the gender (χ2=6.23, p=0.04), duration of breast feeding (χ2=18.6, p=0.04), history of diarrheal disease before polio vaccine administration (χ2=7.7, p=0.02), and stunting (χ2= 114, p=0.00). Conclusion: This study demonstrated that near 90% children achieve seroconversion of OPV and well protected against polio virus. There is an urgent need to focus on factors like duration of breast feeding, diarrheal diseases and malnutrition (acute and chronic) among the children as an immunization strategy.

Keywords: seroconversion, oral polio vaccine, Polio, Pakistan

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154 Engineering of Reagentless Fluorescence Biosensors Based on Single-Chain Antibody Fragments

Authors: Christian Fercher, Jiaul Islam, Simon R. Corrie

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Fluorescence-based immunodiagnostics are an emerging field in biosensor development and exhibit several advantages over traditional detection methods. While various affinity biosensors have been developed to generate a fluorescence signal upon sensing varying concentrations of analytes, reagentless, reversible, and continuous monitoring of complex biological samples remains challenging. Here, we aimed to genetically engineer biosensors based on single-chain antibody fragments (scFv) that are site-specifically labeled with environmentally sensitive fluorescent unnatural amino acids (UAA). A rational design approach resulted in quantifiable analyte-dependent changes in peak fluorescence emission wavelength and enabled antigen detection in vitro. Incorporation of a polarity indicator within the topological neighborhood of the antigen-binding interface generated a titratable wavelength blueshift with nanomolar detection limits. In order to ensure continuous analyte monitoring, scFv candidates with fast binding and dissociation kinetics were selected from a genetic library employing a high-throughput phage display and affinity screening approach. Initial rankings were further refined towards rapid dissociation kinetics using bio-layer interferometry (BLI) and surface plasmon resonance (SPR). The most promising candidates were expressed, purified to homogeneity, and tested for their potential to detect biomarkers in a continuous microfluidic-based assay. Variations of dissociation kinetics within an order of magnitude were achieved without compromising the specificity of the antibody fragments. This approach is generally applicable to numerous antibody/antigen combinations and currently awaits integration in a wide range of assay platforms for one-step protein quantification.

Keywords: antibody engineering, biosensor, phage display, unnatural amino acids

Procedia PDF Downloads 146
153 Genotyping of G/P No Typable Group a Rotavirus Strains Revealed G2 and G9 Genotype Circulations in Moroccan Children Fully Vaccinated with Rotarix™

Authors: H. Boulahyaoui, S. Alaoui Amine, C. Loutfi, H. El Annaz, N. Touil, El M. El Fahim, S. Mrani

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Three Moroccan children fully vaccinated with Rotarix™ have been hospitalized for Rotavirus Gastroenteritis (RVGE) in the pediatric division of the Farabi Hospital, Oujda. Rotavirus G/P genotypes could not be typed because of their delayed crossing threshold (Ct) resolute with a group A rotavirus (RVA) real time RT-PCR. These strains were adapted to cell culture. All viruses replicated and caused extensive cytopathic effects after four or five passages in MA104 cell lines. Significant improvements have been obtained in the amount of viral particles. Each virus multiplied to a high titer (7.5 TCID50/ml). VP7 and VP4 partial gene sequencing revealed distinct genotypes compared to the Rotarix(®) vaccine strain. Two strains were of G2P[4] genotype whereas the third was G9P[8] genotype. Virus isolation while labor intensive, is recommended as a second test, especially when higher sensitivity for conventional RVA genotyping RT-PCR is needed. VP7 antigenic similarities between these strains and Rotarix were determined.

Keywords: esacpe-vaccine, Morocco, Rotarix, G2P[4], G9P[8]

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152 Factors Influencing the General Public Intention to Be Vaccinated: A Case of Botswana

Authors: Meng Qing Feng, Otsile Morake

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Background: Successful implementation of the COVID-19 vaccination ensures the prevention of virus infection. Postponement and refusal of the vaccination will threaten public health, which is now common among the general public across the world. In addition, an acceptance of the COVID-19 vaccine appears as a decisive factor in controlling the COVID-19 pandemic. Purpose: This study's objective is to explore the factors influencing the public intention to be vaccinated (ITBV). Design/methodology/approach: The web-based survey included socio-demographics and questions related to the theory of planned behavior (TPB) and the health belief model (HBM). An online survey was administered using Google Form to collect data from participants of Botswana. The sample included 339 participants, half-half of the participants were female. Data analysis was run using the Statistical Package for the Social Sciences (SPSS). Findings: The study results highlight that perceived severity, perceived barriers, health motivation, and attitude have a positive and significant effect on ITBV, while perceived susceptibility, benefits, subjective norms, and perceived behavior control do not affect ITBV. Among all of the predictors, perceived barriers have the most significant influence on ITBV. Conclusion: Theoretically, this research stated that both HBM and TPB are effective in predicting and explaining the general public ITBV. Practically, this study offers insights to the government and health departments to arrange and launch health awareness programs and provide a better guide to vaccination so that doubts about vaccine confidence and the level of uncertainty can be decreased.

Keywords: COVID-19, Omicron, intention to be COVID-19 vaccine, health behavior model, theory of planned behavior, Botswana

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151 Assessment of Urban Immunization Practices among Urban Mother's in Sri Lanka

Authors: Kasun U. G. Palihakkara

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Although vaccine coverage in Sri Lanka is close to 100%, with the widely spreading vaccine rejection trend reaching South Asian regions, it is essential to catch on whether Sri Lankans are being misinformed from the common misconceptions regarding vaccines. As the rates of target diseases decrease, parents become less accepting of even minor common adverse events. It is essential to preserve the integrity of immunization programs and protect public health by finding out the prevalence of anti-immunization trends. The primary objective of this study was to assess the immunization practices and prevalence of trends related to anti-immunization among urban community in Sri Lanka. A descriptive cross-sectional quantitative study on 323 participants using convenient sampling with 213 self-administered questionnaires. Additionally, 110 online questionnaires were distributed. 31% of the study population does not maintain immunization records for their children. While majority seek information regarding immunization from reliable sources such as the family physician or specialist pediatricians, 30% also refer to unreliable sources such as online communities for their opinion. 31% of study population had not vaccinated for Japanese encephalitis. 73% of the study population had encountered with side effects of vaccination such as fever & 47% believed that such side effects are rare. 52% of the population had hostile attitude regarding the administration of several doses multiple vaccines within a child’s first year. Diseases like polio had been successfully eradicated from Sri Lanka with the help of vigorous vaccination programs. However, majority of the study population believe that there’s no need to keep vaccinating the children for those eradicated diseases and exposing the child for adverse effects of such vaccines. Majority of the population were aware of the existing misconceptions regarding immunization. The most popular misconceptions about vaccines popular among the study population were the MMR (Measles, Mumps, and Rubella) vaccine being a possible cause leading to autism and bowel disease and children getting infected with the disease even after they get vaccinated, may be due to the inactivated vaccine. Disturbingly 22% of the study population believed that vaccines are useless in preventing diseases nowadays. These data obtained from the urban study population reveals that even though statistically Sri Lankan immunization coverage is 100%, there is a possibility of anti-vaccination trend arising in Sri Lanka due to the prevalence of various misconceptions and rumors related to it. Therefore these data recommend the need for thorough awareness among the mothers.

Keywords: anti-vaccination, immunization, infectious diseases, pediatric health

Procedia PDF Downloads 139
150 PCR Detection, Histopathological Characterization, and Autogenous Immunization of Bovine Papillomatosis (Wart) in Cattle, in Mekelle, Northern Ethiopia

Authors: Kidane Workelul, Yohans Tekle, Guesh Negash, Haftay Abraha, Nigus Abebe Shumuye, Yisehak Tsegaye Redda

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Bovine papillomatosis (wart) is one of the economically important bovine skin diseases worldwide, caused by a group of viruses named papillomaviruses (PVs). However, it has often been misdiagnosed as other skin diseases and remained untreated. In order to determine the status of the diseases, twenty-two farms were visited, and fourteen infected cattle with cutaneous papillomatosis were identified from a total of 235. Papilloma biopsies were taken for molecular and histopathological characterization, the therapeutic trial of an autogenous vaccine was evaluated on infected animals. The overall status of bovine papillomatosis in this study was calculated as 5.96% (14/235). The disease was found to be statistically significant in the age groups less than two years (X² = 26.69, P = 0.0001). The more prominent histologically characterized lesions in the sampled tissue were identified as squamous papilloma and fibro-papilloma. The Polymerase Chain Reaction (PCR) based identification revealed that all the clinically and histo-pathologically characterized papillomatosis cases were found to be infected with Bovine Papilloma Virus1(BPV1), indicating that BPV1 was the most common and sole causative agent of the diseases in the study area. In immunizing active bovine papillomatosis, an autogenous vaccine therapeutic trial demonstrated excellent results, with practically full recovery and no recurrence of the infection. Hence, it is concluded that bovine papillomatosis is an economically important disease of young age group cattle as well as a treatable disease. So, the production of marketable autogenous vaccines against bovine papillomatosis should be started and given at an early stage.

Keywords: autogenous vaccine, bovine papillomatosis, bovine papilloma virus1 clinical-pathology, polymerase chine reaction, wart

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149 Identification of Promiscuous Epitopes for Cellular Immune Responses in the Major Antigenic Protein Rv3873 Encoded by Region of Difference 1 of Mycobacterium tuberculosis

Authors: Abu Salim Mustafa

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Rv3873 is a relatively large size protein (371 amino acids in length) and its gene is located in the immunodominant genomic region of difference (RD)1 that is present in the genome of Mycobacterium tuberculosis but deleted from the genomes of all the vaccine strains of Bacillus Calmette Guerin (BCG) and most other mycobacteria. However, when tested for cellular immune responses using peripheral blood mononuclear cells from tuberculosis patients and BCG-vaccinated healthy subjects, this protein was found to be a major stimulator of cell mediated immune responses in both groups of subjects. In order to further identify the sequence of immunodominant epitopes and explore their Human Leukocyte Antigen (HLA)-restriction for epitope recognition, 24 peptides (25-mers overlapping with the neighboring peptides by 10 residues) covering the sequence of Rv3873 were synthesized chemically using fluorenylmethyloxycarbonyl chemistry and tested in cell mediated immune responses. The results of these experiments helped in the identification of an immunodominant peptide P9 that was recognized by people expressing varying HLA-DR types. Furthermore, it was also predicted to be a promiscuous binder with multiple epitopes for binding to HLA-DR, HLA-DP and HLA-DQ alleles of HLA-class II molecules that present antigens to T helper cells, and to HLA-class I molecules that present antigens to T cytotoxic cells. In addition, the evaluation of peptide P9 using an immunogenicity predictor server yielded a high score (0.94), which indicated a greater probability of this peptide to elicit a protective cellular immune response. In conclusion, P9, a peptide with multiple epitopes and ability to bind several HLA class I and class II molecules for presentation to cells of the cellular immune response, may be useful as a peptide-based vaccine against tuberculosis.

Keywords: mycobacterium tuberculosis, PPE68, peptides, vaccine

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148 Proportion and Factors Associated with Presumptive Tuberculosis among Suspected Pediatric Tuberculosis Patients

Authors: Naima Nur, Safa Islam, Saeema Islam, Md. Faridul Alam

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Background: The worldwide increase in pediatric presumptive tuberculosis (TB) is the most life-threatening challenge in effectively controlling TB. The objective of this study was to determine the proportion of presumptive TB and the factors associated with it. Methods: A cross-sectional study was conducted between March and November 2013 at ICDDR-Bangladesh. Two hundred twelve pulmonary and extra-pulmonary specimens were collected from 84 suspected pediatric patients diagnosed with TB based on their clinical symptoms/radiological findings. Presumptive TB and confirmed TB were considered presumptive TB and non-presumptive TB and were isolated by smear-microscopy, culture, and GeneXpert. Logistic regression was used to analyze associations between outcome and predictor variables. Results: The proportion of presumptive TB was 85.7%, and 14.3% of non-presumptive TB. In presumptive TB, vaccine scars, family TB history, and school-going children were 16.6%, 33.3%, and 56.9%, respectively. In contrast, vaccine scars and family TB history were 8.3%, and school-going children were 58.3% in non-presumptive TB. Significant factors did not appear in the logistic regression analysis. Conclusion: Despite the high proportion of presumptive TB, there was no statistically significant between presumptive TB and non-presumptive TB.

Keywords: presumptive tuberculosis, confirmed tuberculosis, patient's characteristics, diagnosis

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147 Surveillance of Adverse Events Following Immunization during New Vaccines Introduction in Cameroon: A Cross-Sectional Study on the Role of Mobile Technology

Authors: Andreas Ateke Njoh, Shalom Tchokfe Ndoula, Amani Adidja, Germain Nguessan Menan, Annie Mengue, Eric Mboke, Hassan Ben Bachir, Sangwe Clovis Nchinjoh, Yauba Saidu, Laurent Cleenewerck De Kiev

Abstract:

Vaccines serve a great deal in protecting the population globally. Vaccine products are subject to rigorous quality control and approval before use to ensure safety. Even if all actors take the required precautions, some people could still have adverse events following immunization (AEFI) caused by the vaccine composition or an error in its administration. AEFI underreporting is pronounced in low-income settings like Cameroon. The Country introduced electronic platforms to strengthen surveillance. With the introduction of many novel vaccines, like COVID-19 and the novel Oral Polio Vaccine (nOPV) 2, there was a need to monitor AEFI in the Country. A cross-sectional study was conducted from July to December 2022. Data on AEFI per region of Cameroon were reviewed for the past five years. Data were analyzed with MS Excel, and the results were presented in proportions. AEFI reporting was uncommon in Cameroon. With the introduction of novel vaccines in 2021, the health authorities engaged in new tools and training to capture cases. AEFI detected almost doubled using the open data kit (ODK) compared to previous platforms, especially following the introduction of the nOPV2 and COVID-19 vaccines. The AEFI rate was 1.9 and 160 per administered 100 000 doses of nOPV2 and COVID-19 vaccines, respectively. This mobile tool captured individual information for people with AEFI from all regions. The platform helped to identify common AEFI following the use of these new vaccines. The ODK mobile technology was vital in improving AEFI reporting and providing data to monitor using new vaccines in Cameroon.

Keywords: adverse events following immunization, cameroon, COVID-19 vaccines, nOPV, ODK

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146 Specific Colon Cancer Prophylaxis Using Dendritic Stem Cells and Gold Nanoparticles Functionalized with Colon Cancer Epitopes

Authors: Teodora Mocan, Matea Cristian, Cornel Iancu, Flaviu A. Tabaran, Florin Zaharie, Bartos Dana, Lucian Mocan

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Colon cancer (CC) a lethal human malignancy, is one of the most commonly diagnosed cancer. With its high increased mortality rate, as well as low survival rate combined with high resistance to chemotherapy CC, represents one of the most important global health issues. In the presented research, we have developed a distinct nanostructured colon carcinoma vaccine model based on a nano-biosystem composed of 39 nm gold nanoparticles conjugated to colon cancer epitopes. We prove by means of proteomic analysis, immunocytochemistry, flow cytometry and hyperspectral microscopy that our developed nanobioconjugate was able to contribute to an optimal prophylactic effect against CC by promoting major histocompatibility complex mediated (MHC) antigen presentation by dendritic cells. We may conclude that the proposed immunoprophylactic approach could be more effective than the current treatments of CC because it promotes recognition of the tumoral antigens by the immune system.

Keywords: anticancer vaccine, colon cancer, gold nanoparticles, tumor antigen

Procedia PDF Downloads 453
145 Barriers and Drivers Towards the Use of Childhood Vaccination Services by Undocumented Migrant Caregivers in Sabah, Malaysia: A Qualitative Analysis

Authors: Michal Christina Steven, Mohd. Yusof Hj Ibrahim, Haryati Abdul Karim, Prabakaran Dhanaraj, Kelly Alexius Mansin

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After 27 years, Malaysia reported polio cases in 2019 involving the children of the undocumented migrants living in Sabah. These undocumented migrants present a significant challenge in achieving the elimination of vaccine-preventable diseases (VPD). Due to the recent polio outbreak among the undocumented migrant children in Sabah, an in-depth interview was conducted among the caregivers of undocumented migrant children to identify the barriers and drivers towards vaccinating their children. Financial barriers, legal citizenship status, language barrier, the COVID-19 pandemic, and physical barriers have been the barriers to access vaccination services by undocumented migrants. Five significant drivers for undocumented migrants to vaccinate their children are social influence, fear of disease, parental trust in healthcare providers, good support, and vaccine availability. Necessary action should be taken immediately to address the problems of vaccinating the children of undocumented migrants to prevent the re-emergence of VPD.

Keywords: Malaysia, polio, Sabah, undocumented migrants

Procedia PDF Downloads 159
144 The Evaluation of Adjuvant Effects of CD154 in a Subunit Vaccine against Classical Swine Fever Virus

Authors: Yu-Chieh Chen, Li-Yun Wang, Chi-Chih Chen, Huy Hùng Đào, Ya-Mei Chen, Ming-Chu Cheng, Wen-Bin Chung, Hso-Chi Chaung, Guan-Ming Ke

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Many recent researches have demonstrated that CD154, a protein primarily expressed on activated T cell molecules, has potentially acted as a molecular adjuvant to improve the immunogenicity of subunit vaccines against viral infections. Classical swine fever (CSF) affects the swine industry worldwide that is one of the most devastating and highly contagious pig diseases. It is listed by the World Organization for Animal Health (OIE) as an infectious animal disease that must be reported. Although pigs vaccinated with subunit vaccines can be differentially diagnosed from those infected animals, subunit vaccines usually need adjuvants to enhance and elicit immune responses. In this study, CD154 was linked with CSFV E2 sequences and then expressed in CHO cells to produce the fusion protein as E2-CD154. The porcine specific CpG adjuvant was also used in one of the formulations. The specific pathogen-free pigs (SPF) at the age of 4-week-old were randomly separated into four groups, vaccinated with E2-CpG, E2-CD154, E2-CD154-CpG or the commercial Bayovac® CSF-E2 vaccine and boosted two weeks after primary vaccination. The results showed that the percentages of CD4+ and CD4+IL2+ in peripheral blood mononuclear cells (PBMC) in E2-CD154 vaccinated piglets seven days after primary vaccination were gained by 1-5% relative to the control group. In addition, the percentages of CD4+IFNγ+ T cells had slightly edged up 0.1-0.3% compared with the control group. Also, increased E2-specific IFNγ levels had edged up CD4+CD8+ T cells found in E2-CD154 and E2-CD154-CpG groups, particularly in the E2-CD154-CpG group. These results implicate that CD154 may enhance cellular immunity and synergistically act with species-specific CpG adjuvant as a dual-phase adjuvant. Therefore, the CD154 may be beneficial as a promising adjuvant in subunit vaccines.

Keywords: CD154, CpG adjuvant, cellular immunity, subunit vaccine, pig

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143 High-Throughput Artificial Guide RNA Sequence Design for Type I, II and III CRISPR/Cas-Mediated Genome Editing

Authors: Farahnaz Sadat Golestan Hashemi, Mohd Razi Ismail, Mohd Y. Rafii

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A huge revolution has emerged in genome engineering by the discovery of CRISPR (clustered regularly interspaced palindromic repeats) and CRISPR-associated system genes (Cas) in bacteria. The function of type II Streptococcus pyogenes (Sp) CRISPR/Cas9 system has been confirmed in various species. Other S. thermophilus (St) CRISPR-Cas systems, CRISPR1-Cas and CRISPR3-Cas, have been also reported for preventing phage infection. The CRISPR1-Cas system interferes by cleaving foreign dsDNA entering the cell in a length-specific and orientation-dependant manner. The S. thermophilus CRISPR3-Cas system also acts by cleaving phage dsDNA genomes at the same specific position inside the targeted protospacer as observed in the CRISPR1-Cas system. It is worth mentioning, for the effective DNA cleavage activity, RNA-guided Cas9 orthologs require their own specific PAM (protospacer adjacent motif) sequences. Activity levels are based on the sequence of the protospacer and specific combinations of favorable PAM bases. Therefore, based on the specific length and sequence of PAM followed by a constant length of target site for the three orthogonals of Cas9 protein, a well-organized procedure will be required for high-throughput and accurate mining of possible target sites in a large genomic dataset. Consequently, we created a reliable procedure to explore potential gRNA sequences for type I (Streptococcus thermophiles), II (Streptococcus pyogenes), and III (Streptococcus thermophiles) CRISPR/Cas systems. To mine CRISPR target sites, four different searching modes of sgRNA binding to target DNA strand were applied. These searching modes are as follows: i) coding strand searching, ii) anti-coding strand searching, iii) both strand searching, and iv) paired-gRNA searching. The output of such procedure highlights the power of comparative genome mining for different CRISPR/Cas systems. This could yield a repertoire of Cas9 variants with expanded capabilities of gRNA design, and will pave the way for further advance genome and epigenome engineering.

Keywords: CRISPR/Cas systems, gRNA mining, Streptococcus pyogenes, Streptococcus thermophiles

Procedia PDF Downloads 257
142 Parental Awareness and Willingness to Vaccinate Adolescent Daughters against Human Papilloma Virus for Cervical Cancer Prevention in Eastern Region of Kenya: Towards Affirmative Action

Authors: Jacinta Musyoka, Wesley Too

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Cervical cancer is the second leading cause of cancer-related deaths in Kenya and the second most common cancer among women, yet preventable following prevention strategies put in place, which includes vaccination with Human Papilloma Virus Vaccine (HPPV) among the young adolescent girls. Kenya has the highest burden of cervical cancer and the leading cause of death among women of reproductive age and is a known frequent type of cancer amongst women. This is expected to double by 2025 if the necessary steps are not taken, which include vaccinating girls between the ages of 9 and 14 and screening women. Parental decision is critical in ensuring that their daughters receive this vaccine. Hence this study sought to establish parental willingness and factors associate with the acceptability to vaccine adolescent daughters against the human papilloma virus for cervical cancer prevention in Machakos County, Eastern Region of Kenya. Method: Cross-sectional study design utilizing a mixed methods approach was used to collect data from Nguluni Health Centre in Machakos County; Matungulu sub-county, Kenya. This study targeted all parents of adolescent girls seeking health care services in the Matungulu sub-county area who were aged 18 years and above. A total of 220 parents with adolescent girls aged 10-14 years were enrolled into the study after informed consent were sought. All ethical considerations were observed. Quantitative data were analyzed using Multivariate regression analysis, and thematic analysis was used for qualitative data related to perceptions of parents on HPVV. Results, conclusions, and recommendations- ongoing. We expect to report findings and articulate contributions based on the study findings in due course before October 2022

Keywords: adolescents, human papilloma virus, kenya, parents

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141 Acute Hepatitis A Outbreak in Men Who Has Sex with Men in a Medical Center in Northern Taiwan

Authors: Yu-Tzu Hsu, Alice Wu, Hsiang-Kuang Tseng

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Introduction: Hepatitis A virus causes acute hepatitis and is usually transmitted by a fecal-oral route of food contamination, which is more prevalent in areas with poor hygienic practices. However, we described a hepatitis A outbreak associated with a fecal-oral route of sexual behavior in men who has sex with men (MSM) in Northern Taiwan. Methods: We retrospectively collected patients with acute HAV infection in MacKay Memorial Hospital, Taipei, Taiwan between July 2015 and November 2016. Demographic data (age, gender, onset time and infection risk), laboratory data (GOT, GPT, bilirubin, HIV status, HBsAg, HCV antibody and syphilis), clinical symptoms and travel history with a foreign tour were analyzed. We compared variables between HIV and non-HIV group. Unless otherwise stated, continuous variables were expressed as mean ± SD, and categorical variables were expressed as number (percentage) for each item. The t test for continuous variables was applied for the comparison between two groups and chi-square for categorical variables were applied for measures of association. Results: We collected 80 cases during the study period. Among them, 54 (67.5%) cases were MSM and 43 (53.8%) cases were HIV positive. The average age was 32.6±7.59 years-old. The average value of initial liver function was 1324 IU/L for AST (GOT), 2100 IU/L for ALT (GPT), and 5.82 mg/dL for bilirubin. We found seven (8.6%) cases were in the status of HBV carrier, five (6.3%) cases were positive for HCV antibody, and 15 (18.6%) cases were co-infected with syphilis. With regards to associated symptoms, 32 (40%) had fever, 46 (57.5%) had nausea, 34 (42.5%) had abdominal discomfort and 46 (57.5%) had general malaise. To compare the non-HIV patients with HIV patients, HIV patients were more likely to be male (p=0.008), MSM (p=0.000), co-infected syphilis (p=0.000) and slowly improving liver function of transaminases (p=0.033, 0.027). Conclusion: The HAV outbreak in Northern Taiwan was mainly occurred in MSM population. Hereafter, our cohort data support a policy in Taiwan to provide one dose of free HAV vaccine shot in this population. Hopefully, the outbreak could be stop by the free vaccine policy and public education.

Keywords: acute hepatitis A, men who has sex with men, human immunodeficiency virus, vaccine

Procedia PDF Downloads 203