Recent advances in nanotechnology has greatly boosted the development of drug delivery systems for therapeutic and vaccine applications, in particular the great success in nanoparticle-based mRNA vaccines for COVID-19. To be noted, rational design and fabrication of safe and efficient nano-carries is the key to lead a successful technology development. It is noteworthy that the delivery performance could be possibly maximized by custom-designed nano-carriers considering the configuration and surface textures of both cargo biomolecules and target cell/environment. Here, we showcase our recent progress on the development of silica based advanced delivery platform.[1-3] Through a biomimetic approach, silica nanoparticles with an intrinsic spiky surface are fabricated and characterized by the unique tool of electron tomography. We demonstrate that control over delicate nanotopography of silica nanoparticles as plasmid DNA vectors has significant impact on the transfection efficacy. A designer spiky of silica nanoparticles acts as hooks to entangle the DNA loops and protect the gene molecules sheltered in the spiky layer against nuclease degradation. Further in vivo demonstrations present enhanced immune responses mediated by spiky silica nanoparticle-mediated DNA vaccines, showing superior performance than the commercial product of in vivo JET-PEI. From bench to market, this spiky nanoparticle based delivery platform is on the industrial translation toward novel nanoparticle-based vaccine technology.
Iron is essential for the functionality of biological processes, however, excess iron is toxic. In humans, due to its toxicity, this metal is found coupled to proteins, such as transferrin, which transports iron to tissues, and ferritin, an intracellular storage molecule of this metal. During an infection, microorganisms obtain iron from the host to survive. Candida albicans, one of the major pathogens responsible for severe fungal diseases, contains a transmembrane protein, iron Ftr1 high-affinity permease, which uses to get iron from ferritin, transferrin, and iron chelators synthesized by other organisms. In the search for new mechanisms to contain fungal multiplication, antibodies are applicable since it is possible to produce specific antibodies against a microbial structure. In this context, the work objective was to produce and perform the biological characterization of egg yolk antibodies (IgY) against the Ftr1 iron permease of C. albicans. A peptide derived from Ftr1 was synthesized and used to immunize laying hens seven times. Eggs from pre-immunized chickens and post-3rd, 5th, and 7th immunizations were collected. IgY antibodies were extracted from egg yolks by the ammonium sulfate precipitation technique. Subsequently, they were purified by molecular exclusion chromatography using Sephadex G-100 resin. The samples containing proteins or peptides were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) at 10%. After this step, the samples were concentrated by filtration using a 50 kDa cut-off ultra centrifugal filter. Then, an avidity enzyme-linked immunosorbent assay (ELISA) was performed to establish the binding strength of the antigen with the antibodies, using urea as a chaotropic agent. To determine the antifungal effect of immunoglobulins extracted after the 7th immunization, in vivo test was performed on larvae of the moth Galleria mellonella, an alternative model of systemic infection. Regarding the results, the production, extraction, and purification occurred successfully, being possible to observe the presence of bands corresponding to the heavy (65 kDa) and light (25 kDa) IgY chains in the electrophoresis gel. Besides, after purification, contaminants (38 kDa) were removed. The immunoglobulin was reactive to the antigen and the avidity was considered low for pre-immunization antibodies, moderate for antibodies extracted post-3rd and 5th immunizations, and high after the 7th immunization. Concerning the challenge in the larvae, at 96 h after the beginning of the experiment, the survival of larvae treated with 80 mg/kg of anti-Fr1 IgY was 90%. In contrast, all larvae that did not receive treatment died (p < 0.0001). Meanwhile, only 16% of the larvae that received 80 mg/kg of pre-immunized IgY survived, and there was no difference between this group and the untreated group (p = 0.2359). This work showed that an antibody produced against C. albicans Ftr1 was able to increase the survival of G. mellonella larvae infected with the yeast.
Vaccination is a key component in the control of animal pathogens that impact the livestock industry. However, deployable, efficacious vaccines remain unavailable for many prevalent diseases, particularly those requiring cell-mediated immunity, such as bovine tuberculosis (TB). In addition,the current ‘vaccinate and challenge’ approach offers little insight into why a candidate vaccine may fail. BCG is a live-attenuated form of Mycobacterium bovis, the causative agent of bovine TB, and is known to induce a strong Th-1 response associated with protective immunity. This project aimed to identify protective signatures associated with cell-mediated immunity in afferent lymphatic DC (ALDC) trafficking from BCG vaccination sites to the lymph node. To achieve this, we utilised a bovine afferent lymphatic cannulation model and collected pseudo-afferent lymph both pre- and 24h post- BCG vaccination, allowing us to perform analysis directly ex-vivo. ALDC were analysed by flow cytometry, single cell and bulk RNA-sequencing. We found three major populations of ALDC draining from the vaccination site: CD172a+ve DC, CD172a-ve DC, and Langerhans Cells. These major populations were divided into 10 sub-populations which exhibited differential gene expression in response to BCG, indicating that individual populations may have differing roles in response to mycobacteria. Pseudo-afferent lymph was also exposed to BCG in vitro, with the aim of using cultured ALDC as a novel way to screen vaccine candidates prior to animal studies.
The aim of this study is the investigation of the polymorphism of the tax gene/Tax protein and pre-miRs-B (pre-miRNA genes of Bovine leukemia virus (BLV)) polymorphism and its relation to Enzootic Bovine Leukosis.
Methods: Peripheral blood samples of 2 to 5 years old female Holstein cattle (farming in Moscow region, Russia) have been used. They have been tested for proviral DNA host genome insertion by PCR (BLV-positive or BLV-negative) and for the presence of antibodies to gp51 BLV antigen by AGID (BLV-seropositive or BLV-seronegative). All samples have been divided into two groups: the group of 16 BLV-seropositive and proviral BLV-positive samples (further: BLV-positive) and another group of 26 BLV-seronegative and proviral BLV-negative samples (further: BLV-negative). WBCs of the samples have been counted on the Abacus Junior Vet 5 Automatic Hematology Analyzer (Diatron, Austria). The BLV-positive samples group has been used for further investigation of the tax gene/Tax protein and pre-miR-B genes’ polymorphism. DNA fragments of tax and pre-miR-B genes have been amplified and further cloned and sequenced. Five or six clones for each of the BLV-positive samples have been sequenced by Sanger’s method. The t-test (http://www.graphpad.com/quickcalcs/) and ANOVA test (GraphPad Prism V.7.04 (1992–2017 GraphPad Software, Inc.)) have been made for statistical testing of the results.
Results: Several alleles of the tax gene and pre-miR-B genes have been received as proof of its polymorphism. Some of them have a highly significant association with an increase and others – with a decreased number of the WBCs in BLV-infected cows. The specific pre-miR-B alleles can be responsible for the increased number of WBCs in BLV-infected cows. The specific tax alleles (and corresponded Tax protein variants) can be responsible for the decreased number of the WBCs in BLV-infected cows, which could be considered the feature of the aleukemic form of BLV infection. They could be named the aleukemic tax alleles.
Introduction and objective: Along with the success of developing biological products of new generation the traditional vaccines, as well as hyperimmune sera, remain as the main means of preventing viral diseases in animals. However, there is a potential threat of the dissemination of emergent infections in a case of insufficient control in the production of animal origin products. It has been already confirmed almost all around the world that pestiviruses are frequent contaminants of live modified vaccines and fetal serum, which induce infection in the organism of animals.
Hobi- like virus or Pestivirus H is one of the 11 main viral species of the genus Pestivirus in the Flaviviridae family. This virus was first identified in 2004 in a batch of bovine fetal serum. This research presents the results of the detection of the Hobi-like pestivirus in the virus vaccine against PPRV used for the vaccination of small ruminants in some farms of the Republic of Tajikistan.
Materials and methods: The studies were carried out in the laboratory of virology of the FSC VIEV. Serum samples of sheep and goats with symptoms of respiratory and reproductive failure from some regions of Republic of Tajikistan were tested by AGID. The vaccine strain of PPRV and strain “Oregon C24V” of BVDV was used as the control antigens. The total RNA was extracted by the commercial kit Syntol (Moscow, Russia) according to the manufacturer’s protocol. RT-PCR for the identification of N and F genes of PPRV and NS3 genes of BVDV was performed in accordance with the recommendations of the OIE Terrestrial Manual, 2013. Viruses were sequenced and phylogenetic analysis carried out using MEGA 7.0.
Results: It has been obtained a positive result when PPRV vaccine strain was used as an antigen. A precipitation line formed between the well with the PPR vaccine and the well with the field serum indicated the presence of antibodies to the PPR virus. In some cases, when examining blood serum samples from sheep and goats with a commercial virus vaccine against PPR, double precipitation lines were observed. In this stage of the study, it has been assumed that the vaccine against PPR contained two viruses. Consequently, we had tested the commercial vaccine used in the Republic of Tajikistan for the presence of contamination. The results of RT-PCR had shown that in addition to the PPR virus, the vaccine contains the genome of bovine viral diarrhea virus (BVDV). When sequencing the amplification products, it was determined that the detected virus belongs to the BVDV genotype 3 or the so-called Hobi-like pestivirus. The sequences of detected virus were deposited to GenBank under accession number KX900607
Nucleic acid-based vaccines represent an attractive alternative to live attenuated and subunit-based vaccines due to their capacity to trigger both humoral and cellular immunity and potential for low-cost and shortened production processes. The fast-track approval and the worldwide vaccination programs for Covid-19 pandemic underscores the efficacy and safety of these novel class of vaccines. DNA and RNA vaccines have been recognized to provide an efficient protection without significant safety risk. Despite the recent successes in Covid-19 vaccines there is strong rationale to continue the investigations of other vaccine strategies to address the need for emergence of new variants (Covid-19), uncontrolled outbreak (flu), the demand for infectious diseases with an unmet need (CMV, RSV), and the requirement for diseases demanding diversified immune response (malaria). Our DNA-based vaccine approach encompasses molecular elements that are designed to improve the breadth of immune response by targeting multiple antigens of a pathogen or multiple mutants of the same antigen, improve the intrinsic adjuvanticity of the vaccine as needed by co-expressing a potent molecular adjuvant particularly against pathogens where conventional vaccines are poorly immunogenic, promote uptake and chemical adjuvanticity of the DNA vaccine with the use of synthetic delivery system, prolong vaccine shelf-life and stability at working temperatures, and utilize an existing cost-effective and scalable process that can be rapidly functional in case of a new outbreak or an evolving existing pandemic. To that end, we have initially produced a family of DNA vaccine vectors expressing one or more of SARS-CoV-2 surface antigens as a proof-of-concept target, verified vector composition, and demonstrated expression of the encoded genes. We have also developed an intramuscular vaccine formulation based on a covalently modified co-polymer that yields high levels of a reporter gene expression within 24 hours after treatment, a critical time window for immune activation, followed by durable expression potentially conducive to B- and T-cell responses. Immunization of Balb-C mice with a plasmid expressing the Spike protein of SARS-CoV-2, resulted in the production of IgG antibodies with evidence of viral neutralization and cytotoxic T-cell response specific to the antigen. Parallel in vivo studies are in progress to optimize vector and antigen design, improve the delivery system, explore alternative route of administration, identify the optimal vaccine dose and regimen. The current data and results from the ongoing studies will be presented at the conference.
We will present several examples to highlight the capabilities of the fluorescence A-TEEM method, a novel approach which combines UV/Vis with fluorescence EEM spectroscopy. The benefit of this approach is the ability to provide robust and rapid spectroscopic characterization of vaccine components and vaccine formulations, samples that are extremely challenging to traditional spectroscopies, such as Raman and NIR. The Coronavirus pandemic applied historic pressure on vaccine development and production, collapsing development timelines from years to months, with many firsts in formulation and production. Requirements for product quality still had to be met though, highlighting the need for rapid analytical techniques to characterize vaccines from R&D to formulation development, and through manufacturing to final QA/QC. Spectroscopic techniques are known to be rapid and are therefore used extensively for PAT and QA/QC testing. However, the “go-to” techniques such as Raman and NIR often do not work for vaccines, as they struggle with low protein concentrations typical of these formulations. The fluorescence A-TEEM method is a unique alternative, combines high sensitivity and specificity, with limits of detection to 0.015 ug/mL, and data acquisition times typically under 60 seconds. As the method combines two well established techniques (UV/Vis and fluorescence), the technique can be validated by following the relevant USP chapters, <853> and <857>. We will present results from an A-TEEM study on four closely related combination vaccine formulations, Solo-Jec brand canine vaccines from Boehringer Ingelheim VetMedica. A-TEEM was able to accurately differentiate between the four formulations, even when they differed by only a single coronavirus component. To ensure repeatability, calibration data (two separate samples for each formulation) were collected, and reproducibility was assessed with a third unique set of validation samples, collected on a different instrument by a different operator. The A-TEEM is able to identify and validate “unknown” samples with 100% certainty. In addition to vaccine formulations, we will present AAV characterization studies, where the A-TEEM was able to rapidly (<90sec) differentiate biotinylated AAV samples that are complexed with streptavidin-dye conjugates from uncomplexed AAVs.
In this theme the author will discuss various explanations that was used or not to implement this kind of rules in Europe and in many countries nowadays.
Purpose; to demonstrate that using only this type of measures to control the spread of the virus, will not be reducinf the incidence of he infecton because the author is demonstrating that the type of population that we have nowadays are not immune competent but immune deficient in energy , that will compromise the formation of antibodies for SARS-CoV-2 after receiving the vaccines and also, there are other variants and strains that only one vaccine cannot prevent all forms of SARS-CoV-2. Also, there are studies showing that persons that are fully vaccinating can spread virus even if assymptomactic due to maitaing virus in the nose and in the throat.
Methods: In this study,the author is showing the studies that we have nowadays reagarding this theme and also, said by many specialists in the area.
Results; in these studies, they are waiting for the B and T cells responses after the vaccination. In another talk from Antony Falci, he is also saying that there is no evidence that the vaccine can control the wide spread of the virus. And there will be increase in the formation of auto-immune disease in the near future, according to some studies , due to this massive vaccine implementation.
Conclusion: the conclusion of this study is that the implementation of universe vaccination is not based on studies very well done and are based on studies that have no conclusion yet. According to the author, the vaccination will not control the widespread of the SARS-CoV-2 infection due to the fact that persons full vaccinated can spread virus to other even if assymptomactic and the majority of the population nowadays are with immune supressed, induced by the chronic exposition to electrtomanectic waves and affecting our energy and immune system, leading to less respónse to vaccines nowadays. The use of other forms of measurements to increase the immune system of the entire population nowadays, such as the use of highly dilluted medications such as homeopathies, increasing the viotal energy of the population , that is very low nowadays, is the major importance, to treat the cause of the problem and not just treating the symptoms, that is the SARS-CoV-2 infection.
This process is not very easy to explain in the eyes of Western medicine and the author will explain it from the perspective of traditional Chinese medicine, following the commandments of Hippocrates, father of medicine. In an article written by the author entitled Is SARS-CoV-2 Strong or Our Body Is Weak? it shows that more than 97% of patients have low Zheng-Qi, which is the energy that protects the individual's body against the invasion of external pathogenic factors. In another article written by the author entitled Energy Alterations and Chakras' Energy Deficiencies and Propensity to SARS-CoV-2 Infection, she demonstrates that more than 90% of her patients studied between 2015 and 2020 are without energy in the five massive internal organs , which are responsible for maintaining our health, and that this energy deficiency is responsible for the formation of infectious and non-infectious diseases today. In another article written by the author, entitled What have behind in all kinds of infections that we need to know?, the author says that what all bacterial, viral and fungal infections have in common, is the deficiency of energy in the chakras and formation of internal Heat.
The purpose of this study is that, the author wants to show that most people are considered immunedefiient , due to the energy deficiency pattern , generating in this way, internal Heat formation, this being the predominant factor for colonization and infection by bacteria and viruses.
Method: the author uses many articles written by her explaining how to treat community and nosocomial infections withouyt need to use any antibiotics or antiviral medications.
Results: because she knows that If we replenish the energy of these patients in order to reduce the formation of internal Heat, there would no longer be viral colonization in the nasal cavity of these vaccinated individuals, because currently, even if they are immunized, the underlying cause of immunodeficiency was not treated, which is low state os energy, due to the electromagnetic radiation generated by the modernization of communication.
Conclusion; to reduce virus colonization even after vaccination for COVID-19, individuals need to improve their energy state, which is weakened, to reduce the production of internal Heat, responsible for the adherence of bacteria and viruses in individuals with these infections and which, according to the author's experience, who is a specialist in infectious diseases, but treats most community and hospital diseases without using antimicrobials, she uses the methods of older medicines, such as Chinese medicine and thus manages to treat most of the infections by resistant bacteria and viruses, only drawing internal Heat and rebalancing the internal energies of Yin, Yang, Qi and Blood.
StreptInCor, a candidate vaccine against S. pyogenes is based on protective 55 amino acids residues of C-terminal portion of the M protein. Experimental assays have demonstrated that the StreptInCor peptide induces high titers of opsonic and neutralizing and protective antibodies in outbred immunized mice. Using HLA class II transgenic mice, it was possible to evaluate the immunogenicity and safety of the StreptInCor vaccine epitope for a period of one year. Specific and non-auto reactive antibodies were produced as well as no autoimmune or pathological reactions were observed in the heart or other organs of these animals. We also performed several studies in mini-pigs in order to evaluate the immune response and safety by submitting these animals to echocardiogram examination before immunization and after the four doses treatment. No alterations were observed. In addition, both repeated intramuscular-dose toxicity tests (28 days) with four doses and echocardiography procedure in mini pigs after 28 days were performed. No harmful effects to the tissues and organs studied were observed indicating that the vaccine is safe. StreptInCor vaccine also induces regulatory T cells (Treg) that strongly indicate that the vaccine peptide may have therapeutic potential to control both inflammatory and autoimmune response in RF/RHD patients.
There is no doubt that vaccines are the most effective solution against the global challenges that pandemic brought to the world. In addition, thousands of studies all around the web are publishing the succeed results and social media is spreading them rapidly. Nevertheless, there are some noticed cases about people whose general condition turned into worse after the first or even second dose of vaccines. During my personal practical experience in admission department and infectious diseases department of several city hospitals I worked, there are observed cases about patients who had complications with their general condition after they were vaccinated with vaccines such as Sputnik-V, Hayat-vax, Sino-Vac and others. This study is aimed to appeal for a careful attention of society to vaccines with people with chronic and acute infection diseases, or people with first registered personal complications after vaccination and people of elder ages. Also, for an additional information, some clinical cases and patients’ histories of diseases will be shown as an example to expand the understandings and widen the view of contraindications and actual side effects which might be already known but underestimated properly or some instances that could be marked as new or rare. It was noticed that during the survey many patients with complications reported they were not informed about possible side effects that may happen after vaccination and were not asked if they have or not other chronic diseases already. So, it is quite clear there is a need of common general list of questions that should be taken as a strict rule by medical personnel. Despite the fact it is not the first year of pandemic, there is still part of people who is not informed well or informed incorrectly which means it is a call for Health Department to increase the methods of spreading the knowledge in all modern and simple ways for public.
Lassa fever (LF) is an acute viral haemorrhagic disease caused by Lassa virus (LASV), which is endemic in Western Africa and is associated with high rates of infection and mortality. Currently, there are limited treatment options and no licensed vaccines to protect against LASV infection. Previously, we reported on a DNA vaccine, INO-4500, encoding the LASV (Josiah strain) glycoprotein precursor (LASV GPC) gene, that elicits protective immunity and completely protects guinea pigs and non-human primates against viremia, illness, and death after Lassa virus exposure. Here, we present T cell responses after vaccination with INO-4500 from a randomized, placebo-controlled phase 1 LASV vaccine trial in healthy adults. Two doses of 1 mg of INO-4500 were administered by intradermal injection followed by electroporation using CELLECTRA® 2000 device at day 0 and week 4. The cellular immune response was assessed at day 0 and weeks 6, 12, 24 and 48 following vaccination. INO-4500 elicited a strong T cell response against overlapping LASV peptide pools as measured by IFNg-ELISpot compared to placebo controls. Using flow cytometry, we examined specific LASV reactive T cells after stimulation with LASV peptides and analysed their cytokine profile. Significantly higher frequencies of polyfunctional CD4+ and CD8+ T cells, producing IL-2, IFNg and TNFα, were observed at week 6 and 12 following vaccination. Also, activation-induced LASV-specific IRF4+CD137+-CD4+ and -CD8+ T cells were significantly expanded at week 6 but were detectable up to 48 weeks after vaccination, suggesting induction of long- term memory antigen-specific T cells. This is supported by a significant increase of IL-2, IFNg, TNFα, IL-6, IP-10 and MIG in the supernatant of cultured cells stimulated with LASV peptides at different time points post-vaccination as detected using Legendplex. These results revealed that INO-4500 LASV-GPC DNA vaccine induce robust T cell responses, which are critical for protection against LF. The activation and persistence of LASV-specific T cells supports the potential of INO-4500 to protect against LASV.
To prevent next flu pandemics and to reduce the burden of seasonal flu infections, there is an urgent need to develop vaccines that are broadly protective against different influenza strains. In this regard we investigated polymeric nanoparticles as antigen carriers of a highly conserved epitope, the extracellular domain of M2 membrane protein (M2e) of the influenza type A virus, and compared immunogenicity from simple adsorption versus the chemical conjugations of M2e on to polymeric nanoparticles. We synthesized two types of nanoparticles from similar molecular weight polymers: i) PmNPs: nanoparticles made of poly (lactic-co-glycolic acid)–polyethylene glycol with maleimide linker, ii) and PNPs: nanoparticles made of methoxy-polyethylene glycol-poly (lactic-co-glycolic acid). PmNPs and PNPs were characterized by transmission electron microscopy and dynamic light scattering. We conjugated M2e on the surface of PmNPs, via maleimide-thiol rxn, whereas for PNPs, M2e was physically adsorbed at the same dose. We were immunized mice via intramuscular or intranasal routes on day 0 and 21 using M2e conjugated PmNPs or PNPs with soluble form of CpG as an adjuvant. Anti-M2e-specific IgG, IgG1 and IgG2a responses were determined in serum samples and lungs lavage after day 42 by ELISA. Cellular immune response was determined through cytokine analysis from splenocytes restimulation. Mice were challenged after day 42 with 3xLD50 of various strain of influenza virus and survival and weight loss were monitored daily. We synthesized PmNPs and PNPs of the comparable size (~100 nm). We found that covalently attached M2e at PmNPs with CpG had a more significant immune response and protection with challenge than from physically adsorbed M2e on PNPs. The intramuscular route was identified to be significantly better than the intranasal route in terms of anti-M2e immunoglobulin G (IgG), IgG1 and IgG2a, cellular immune response and protection with challenge. We found full protection in PmNP-M2e with CpG immunized mice against challenged influenza viruses such as A/California/04/2009 (H1N1pdm), A/Victoria/3/75 (H3N2) and A/PR/8/34 (H1N1). We equally found reduced lungs virus titers and histopathological signs at day 5 after challenge in PmNPs-M2e with CpG immunized mice in comparison to other groups. Based on these data, we are successfully demonstrated that the use of PmNPs-M2e with CpG formulation could lead to the development of a universal influenza vaccine.
‘History teaches us all about the course of past pandemics and the importance of the use of effective and safe vaccines to contribute to prevent diffuse bacterial and viral infections. But vaccines have also generated ethical controversies over the years. It is important to learn from the past and to make sure the efficacy and the safety of the available and future vaccines is continuously and carefully monitored.
There is an urgent need today to merge all serious and non-serious adverse events post COVID-19 vaccine administrations generated on a national and continental level, in order to globally assess the safety profile of these vaccines, so they can be conveyed in one only global clinical safety database. This harmonized and coordinated effort would allow generating a global clinical safety report to be continuously assessed and monitored by an independent global panel of safety experts across the world. The approach is of highest clinical relevance to figure out any potential safety signal emerging from all adverse events around the world post COVID-19 vaccinations. The approach would allow to eventually recalibrating, if deemed necessary, the clinical positioning and the labelling of the approved vaccines, which are currently utilized for preventing COVID-19 infection.
This abstract aims to generate awareness on this topic and to invite all involved regulatory authorities to collate all the adverse events arisen from every single country and to promptly disclose in one global report the available information related to the serious and non-serious side effects occurred post-COVID-19 vaccinations. This is of clinically high relevance, particularly to the whole world community. It is extremely important to carefully evaluate and not to downplay any emerging safety signal from the real world data, even if this was assessed as ‘minimal’.
Due to the accelerated timing to develop and to produce the approved COVID-19 vaccines by several regulatory agencies, also by considering the limited information from the available clinical studies at the time of their approval, i.e. because of the studies sample size; the limited number of patients/subgroups; the study durations, the absence at that time of the latest variants of the virus etc. based on prediction of more vaccines to become available, the clinical relevance to urgently identify vaccine-specific subgroups of people at higher risk to develop specific side effects, especially those most serious ones, all this is of high clinical importance. The approach could help to better understand the use of the existing vaccines and the most eligible populations needed to be vaccinated and to complete the full course of each vaccination. This is also relevant in light of future vaccination recalls and a full global vaccination campaign necessary to tackle the ongoing pandemic’.
Toll-like receptors (TLRs) are essential for activation of innate immunity and initiation of adaptive immune response. They have been associated with several respiratory diseases, but information on the relation of TLRs with SARS-CoV-2 still needs to be completed. Endosomal TLRs can detect viral or bacterial nucleic acids and have an important role in initiation of immune responses against the pathogens. TLR7/8 can detect consecutive uridine-containing single-stranded RNA of the viral genome. TLR9 binds CpG signaling motifs in bacterial and viral DNA molecules, and is found to be highly expressed in peripheral blood mononuclear cells after infection with SARS-CoV. Also, the SARS-CoV genome has higher numbers of TLR9 signaling motifs than some other respiratory diseases viruses. It was shown that SARS-CoV-2 genome has more TLR7/8-detectable motifs than SARS-CoV genome, representing a higher probability to interact with TLR7/8. This was linked to the potential ability of SARS-CoV-2 to induce pro-inflammatory responses in severe lung injury cases. Since it was shown that alpha and delta variants are associated with higher transmission rates, in this study, we performed a bioinformatic analysis on the interaction of endosomal TLRs and SARS-CoV-2 wild type, alpha and delta variants. We used Sequence Searcher software for searching TLR7/TLR8 RNA motifs, and TLR9 CpG motif in the SARS-CoV-2 genome of Wuhan reference sequence and alpha and delta variants. Data analysis showed that the overall number of genomic motifs detectable by TLR7/8/9 in SARS-CoV-2 alpha and delta variants was lower than in SARS-CoV-2 wild type sequence. This suggests that the endosomal TLRs may have no important effect on higher transmissibility of SARS-CoV-2 alpha and delta variants. Also, lower detectable motifs in delta than alpha variant, indicated a potential lower binding affinity to endosomal TLRs compared with alpha. As a conclusion, higher transmission rates reported for alpha and delta variants may not be due to the genomic mutations that alter the number of TLR7/8/9-detectable motifs.
Audience Take away:
Mycobacterium tuberculosis is one of the most dangerous pathogens. Bacterial resistance to antituberculosis drugs grows each year, but searching for new drugs is a long process. Testing for available drugs to find active against mycobacteria may be a good alternative. In the presented study, we used the fast?growing and nonpathogenic Mycobacterium smegmatis to investigate the effect of aureolic acid group antibiotics.
In this work, M. smegmatis mc2 155 strain was used. To antibacterial activity analysis, Olivomycin A, Mithramycin A, Chromomycin A3, BRACO?19, and TMPyP4 (all from Sigma?Aldrich, Missouri, USA) (Supplementary Materials Figure S1) were added to the cultures, at a final concentration of 10 μM. As a positive control, Kanamycin was used at a final concentration of 20 μM. Negative control samples were treated with the same volume of DMSO (a solvent for all chemicals above). G4 motifs in the genomes of M. tuberculosis and M. smegmatis were analyzed, and the ability of the aureolic acid group drugs to stabilize G4 motifs was tested. For scanning electron microscopy (SEM), fixed cells examined using a scanning electron microscopy multipurpose analytical complex Merlin (Carl Zeiss, Germany). Transcriptomic analysis was conducted on Illumina HiSeq 2500.
We presumed that antibiotics of this group may be potential G4 ligands. However, this was not confirmed in our analyses. Our data demonstrate antimycobacterial activity of Olivomycin A. We showed that it significantly inhibits the growth of M. smegmatis, the closest relative of M. tuberculosis. Transcriptomic analysis revealed a decrease in the transcription of several essential genes and an active cell response on the stress. Mycobacterial cells cultivated in the presence of sublethal doses of antibiotic (0.5 μM) were elongated and formed conglomerates not typical for control cells. Transcriptomic analysis documented increased expression of MSMEG_3743/soj and MSMEG_4228/ftsW, involved in cell division. Therefore, drugs may affect cell division, possibly disrupting the function of the Z?ring and the formation of a septum. Additionally, a decrease in the transcription level of several indispensable genes, such as nitrate reductase subunits (MSMEG_5137/narI and MSMEG_5139/narX) and MSMEG_3205/hisD was shown. We concluded that the mechanism of action of aureolic acid and its related compounds may be similar to that bedaquiline and disturb the NAD+/NADH balance in the cell. All of this allowed us to conclude that aureolic acid derivatives can be considered as potential antituberculosis drugs.
Rheumatoid arthritis (RA) is one of the most common chronic inflammatory autoimmune disease. Patients with RA have a significant increased risk of serious infection following treatment with disease- modifying antirheumatic drugs, and clinical guidelines strongly recommend pneumococcal and influenzae vaccines; recurring allegations on their association with disease incidence are raised. The role of vaccination on the occurrence of flare-ups of the disease is not fully known. Associations with vaccines have been reported, such as those against tetanus, hepatitis B, and influenza, but causality has not been confirmed. This study aimed the association between vaccination and the occurrence of acute flare-ups in patients diagnosed with RA A case-crossover study was conducted within a cohort of SLE. All patients with a diagnosis of RA were identified between 1st Jan. 2008 and 31st Dec. 2018 in a nationwide linked health care database covering 97% of the French population. A RA flare-up was defined as either a new pharmacy dispensing claim for high-dose corticosteroid or a hospitalization with a RA-related primary discharge diagnosis (ICD-10 codes). Vaccine exposure in the 2 months prior to the date of flare-up (risk window) was compared to prior exposure in up to 4 control time windows per patient (each of 2 months). GEE models were used to account for the occurrence of multiple flare-ups within an individual, adjusting for health care utilisation. Stratification by type of flare-up (first flare-up in incident patients or any flare-up in prevalent patients) and by type of vaccine (bacterial or viral) was conducted.A total of 223,612 patients with RA were identified. The mean age of patients at study entry was 56.1 years [SD:14.8] and 72.6% were females. A total of 799,634 acute RA flare-ups were identified over the study period, among which 79,417 (9.9%) were first episodes in incident RA patients. Overall, 81.9% RA patients were vaccinated at least once during the study period, distributed as follows (non-mutually exclusive): vaccine combinations (46.1%), flu (53.0%), pneumococcus (41.7%), hepatitis B (2.4%),tetanus (5.9%) and others (<1%). In patients with prevalent flare-ups, 57,218 (7.49%) vaccinations occurred during the risk window compared to 168,234 (7.11%) occurring in the control windows (odds ratio (OR): 1.13; 95% confidence interval (CI): [1.12-1.14]). The OR for viral vaccines was OR=1.19 [1.18 - 1.21]), for bacterial vaccines OR=0.96 [0.94 – 0.98] and for combinations: OR= 0.93 [0.91-0.96]. Findings were similar for incident flare-ups. From this large-scale study, we observed a small association between vaccination and flare-ups in RA patients. Further research is needed to confirm this association notably the role of circulating viral epidemics and disease activity on vaccination and flare-ups.
Immunoinformatics plays a key role in vaccine design and antibody production. In the past, antibody design and vaccine development are expensive and time-consuming. Nowadays, advances in the field of bioinformatics have provided practical tools that can be used to lessen the time and cost of vaccine and antibody design. The Immunoinformatics approach allows the identification of the immunogenic epitopes from the pathogen genomes. Also, the ideal and immunogenic parts could be developed as potential vaccine candidates to trigger protective immune responses in the hosts. In our studies, are identified the potential and immunogenic epitopes of a selected protein by bioinformatics tools. Then, these are selected based on allergenicity, antigenicity, and solubility features. And we use linkers and adjuvants to structure the epitope vaccines. Then, the 3D structure of the protein was predicted, and its affinity to different HLA, TLR was investigated by molecular docking. These methods have been performed with a wide range of bioinformatics tools to design vaccines against Covid19, Acinetobacter Baumannii, and Neisseria meningitidis. The design technique of these vaccines will be clearly mentioned in the lecture.
Major hindrances to getting a COVID-19 vaccine include vaccine hesitancy, skepticism, refusal, and anti-vaccine movements. Several studies have been conducted on attitudes of the public towards COVID-19 vaccines and the potential influencing factors. The purpose of this scoping review is to summarize the data available on the various factors influencing public attitudes towards COVID-19 vaccination. This scoping review was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for Scoping Reviews (PRISMA-ScR) Statement. PubMed, Embase, Web of Science, and Cochrane Central were searched without restrictions to reclaim all publications on the factors that shape individuals’ attitudes towards COVID-19 vaccines from 1 January 2020 to 15 February 2021. Fifty studies were included. The scoping review revealed that the factors influencing public attitudes towards COVID-19 vaccines were embedded within the different levels of the socio-ecological model. These factors included the sociodemographic characteristics of the individuals, individual factors, social and organizational factors. In addition, certain characteristics of COVID-19 vaccines themselves influenced public attitudes towards accepting the vaccines. Understanding various population needs and the factors shaping public attitudes towards the vaccines would support planning for evidence-based multilevel interventions in order to enhance global vaccine uptake.
Background – Since the day the Novel SARS Cov has been detected and the ensuing pandemic the search for a cure or prevention has been the only target of the medical fraternity. As the second wave racked havoc Vaccine seemed the only viable option to stop this global surge. WHO & subsequently Government of India have issued emergency use authorization to 2 vaccines. Our study is aimed at estimating the seroconversion rates and to identify predictors of antibody titres in vaccinated healthcare workers in VIMSAR, Burla.
Methods – This is a part of the ongoing repeated cross-sectional study. Participants were enrolled well above the Sample size (322) to increase precision. Two rounds of survey were conducted & is being reported. Serum IgG antibodies against spike protein of SARS-CoV-2 was estimated using Elecsys® Anti-SARS-CoV-2S is an immunoassay by ECLIA based Cobas e411 analyzer. Univariate and multivariate regression were used in statistical analysis.
Results - Our results show 95.1 % and 99.5% of the vaccinated individuals have developed anti spike protein antibodies after the 1st and 2nd dose respectively. Previous Covid infection was Significantly Correlated with antibody production and age was negatively correlated. No difference was reported for sex, occupation, diabetes.
Conclusion. Our interim analysis report is coherent with the available literature and research regarding high efficacy of Covid vaccine as far as seroconversion is concerned.
Brucellosis is a zoonotic bacterial infection of pandemic potential – caused by Brucella species. Lack of vaccines against human brucellosis despite continuous research has led us to address this issue using several selectively chosen antigens pertaining to Brucella spp. We have been utilizing different marketed adjuvants e.g. Aluminium hydroxide, MF-59 etc. Simultaneously, we have been developing nano-encapsulations of various antigens namely – rL7/L12, rOmp25 and rOmp28 etc.
Poly-lactide-co-glycolide (PLGA) has been one pf the most successful with all the antigens mentioned above in eliciting a good humoral immune response as well as causing a huge reduction in the organs’ bacterial load especially liver and spleen. Additionally, we have been using nano-liposomes which were similar in their efficiacy aspects but had much inferior stability upon storage.
Post-immunization, several parameters e.g. IgG antibody levels, cytokines – their levels, and bacterial load in the organs and all the antigens being highly immunodominant produced significantly high antibody titers apart from ensuing varied cytokine release patterns.
To summarise, Omps were found to spike apical immune responses as well as maximal protective efficacy. Thus, Omps hold a huge promise as prophylactics of future use. More research is aimed to be conducted to understand the immune memory pertaining to these antigenic formulations.
During recent years we have witnessed numerous occurrences of viral infectious diseases such as Ebola, MERS, SARS, and recently COVID-19 with a drastic negative effect on human health. Following the outbreak of the COVID-19 pandemic, it has been declared a public health emergency of international concern by the World Health Organization. The causative agent of this infection would be able to transmit via contact with contaminated secretions of infected people, or close contact with a cough or sneeze. Prompt development and comprehensive investigations about the production of an efficacious vaccine against the SARS-CoV-2 have been performed in some countries, but rigorous studies are required to determine the safety and potency of candidate vaccines. SARS-CoV-2 vaccines are being developed using several different platforms. Some of these are traditional approaches, such as an inactivated virus or live attenuated virus platforms, some are newer approaches, such as recombinant proteins and vector vaccines, and some have never been previously employed in a licensed vaccine, such as RNA and DNA vaccines. Also, following availability and widespread uptake of SARS-CoV-2 vaccines, efficacy issues that were not addressed in clinical trials will need to be evaluated, including the duration of protection and the potential need for additional doses, effectiveness in subpopulations not included in trials, and impact on community transmission. Also, in my country some new vaccines were produced as a new effective vaccine by several Research Institutes as first time in Iran. In this lecture, some COVID-19 vaccines will be discussed for effectiveness, safety, protection and efficacy in sensitive and high risk population.
The physicians are talking about the treatment of fever indifferently. They are talking about not to treat fever but for the underlying cause of fever. At the same time when people have a fever, they instruct to reduce fever urgently. The cause of fever and cause of disease both are different. The physicians misunderstand the cause of disease as cause of fever. If we remove the cause of disease the fever never cures. If we remove the cause of fever, it can be immediately cured. The basic cause of fever is increased severe inflammation and decreased blood circulation. If we remove the cause of disease, the disease will not be cured.If a worm eats the stem and leaf of a plant, we can kill the worm with pesticides, then the destroyed part of the plant will not recover completely. Likewise, we can kill some kind of bacteria with antibiotics. But the problems made by bacteria will not be resolved.The actual treatment to cause of fever.The actual treatment to fever is to increase blood circulation. Two ways to increase blood circulation. 1. Never allow body temperature to lose 2. Apply heat from outside to the body. When the temperature produced by the body due to fever and heat which we applied on the body combines together, the blood circulation increases.
Then the body will stop to produce heat to increase blood circulation. And the body will get extra heat from outside without any usage of energy.
How can we prove that the cause of fever and cause of disease both are different?
“If we ask any type of question-related to fever by assuming that the cause of fever and cause of disease both are different, we will get a clear answer. If we avoid or evade from this definition, we will never get a proper answer to even a single question
If we do any type of treatment by assuming that the cause of fever and cause of disease both are different, the body will accept the cause of fever, at the same time body will resist whatever treatment to decrease temperature and blood circulation.No further evidence is required to prove the cause of fever and cause of disease both are different.
To generate more effective subunit vaccines against complex organisms like Leishmania parasites, we need to fuse several immunogenic proteins together. However, the critical issue is that which arrangement of the constituting components should be selected as the favored combination for vaccine design. These days, RNA and protein structure analysis techniques together with immunoinformatics are the handiest approaches available for selecting among the appropriate combinations. To advantage this approach, all possible combinations are designed and analyzed at mRNA and protein levels. At the mRNA level, the full sizes of the transcripts are estimated based on the used expression vector. Then, the secondary structures of mRNAs are predicted by specific web servers. At the protein level, the 3D structures of all combinations are modeled by online platforms. Then, the predicted 3D models are refined and validated by different parameters. Eventually, validated models are superimposed to the original individual proteins to find out structural identity, the higher the similarity the better the reliability of candidate combination. On the other hand, the combined structures are also further analyzed for junctional epitopes by immunoinformatics. This approach was used to combine two immunogenic salivary proteins (PpSP15 from Ph. papatasi and PsSP9 from Ph. sergenti) together to develop an effective vaccine against cutaneous leishmaniasis. The best combination as the vaccine candidate was selected based on mRNA and protein stability results besides peptide analysis and will be presented as an example model for a multi-protein vaccine design.
Japanese encephalitis (JE) is a vector-borne zoonotic viral disease caused by (Japanese encephalitis virus, JEV). Vaccination is the most effective way to control JE in both humans and pigs. However, JEV genotype shift that the dominant genotype III (GIII) has been replaced by genotype I (GI) raised concerns about the effectiveness of GIII-derived vaccines against the GI strain infection. Indeed, GIII-derived vaccine showed a reduced protection against GI strain challenge, suggesting a potential need of development of GI-derived vaccine. In this study, a comparative analysis of the phenotypic and genotypic properties of an attenuated GI strain (SD12-F120) and its virulent parental strain (SD12) was performed. To characterize the attenuated GI strain SD12-F120, the phenotypic and genotypic characteristics of SD12-F120 with its virulent parental SD12 strain were compared in vitro and in vivo. SD12-F120 formed smaller plaque on BHK-21 cells and showed the reduced replication in mouse primary neuron cells and mouse brains as compared with SD12. Mice inoculated with SD12-F120 up to 105 PFU via either intraperitoneal or intracerebral route showed no clinical signs of JEV infection, indicating highly attenuated phenotype in terms of both neuroinvasiveness and neurovirulence. Immunization of mice with SD12-F120 provided a complete protection against SD12 challenge. Comparison of genome variations between SD12-F120 and SD12 revealed that SD12-F120 harbored 29 nucleotide variations, of which 20 were considered silent nucleotide mutations, while 9 resulted in eight amino acid substitutions: two in E, one in NS1, two in NS3, one in NS4B and two in NS5 proteins. Comparison of the amino acid variations of SD12-F120 vs SD12 pair with those from other four isogenic pairs of the attenuated and their virulent parental strains revealed that the numbers and positions of amino acid variations were different from each other. Out of the five pairs, the variations at E138 and E176 positions of E protein were identified in four and three pairs, respectively. The remained amino acid variations were almost unique to their respective strain pairs, suggesting that the genetic changes acquired during the attenuation process were likely to be strain specific and that the mechanisms associated with JEV attenuation/virulence were complicated. The SD12-F120 was obtained by serial passage of its virulent parental SD12 strain on BHK-21 cells, which was not appropriated for vaccine production. Therefore, SD12-F120 was further passaged on Vero cells for 20 passages and a Vero cell-adapted strain SD12-F120VC was obtained. SD12-F120VC had an increased in virus titers compared to early passages, showing an adaptation to Vero cells. The plaque morphology of SD12-F120VC was same as that of SD12-F120. The animal experiments showed that SD12-F120VC had attenuation phenotype in suckling mice. Vaccination of mice with SD12-F120VC completely protected vaccinated mice against challenge with SD12 strain, but failed to provide the vaccinated mice complete protection against the challenge of GIII N28 strain. The neutralizing antibodies titer of immunized mice against SD12-F120VC and SD12 was higher than that of heterologous N28 strain. SD12-F120VC harbored 6 amino acid substitutions in prM protein, of which 5 were present in the pr domain of prM. These amino acid substitutions may be involved in the adaptation of SD12-F120 to Vero cells. Overall, the phenotypic and genotypic characteristics of SD12-F120 with its virulent parental SD12 strain were compared in vitro and in vivo, and the protection of the attenuated strains was determined in mice. The outcome would be useful for development of GI-derived vaccine.
This fact is well known that the appearance and development of all fish disease process could be the result of the interaction between three parameters such as pathogen, host, and environment. Therefore, only multidisciplinary studies involving knowledge of the characteristics of the potential pathogenic microorganisms for fish, aspects of the biology of the fish hosts, as well as a better understanding of the environmental factors affecting them, will allow the application of adequate measures to prevent and control the main infectious diseases limiting the production of freshwater and marine fishes. Vaccination is becoming an increasingly important part of aquaculture, since it is considered a cost effective method of controlling different threatening diseases. Also, it was alternated for Antibiotic application in some developed countries and it could be positive aspect to decreasing antibiotic resistance in some familiar pathogens in the environment and aquatic ecosystems. The term vaccination strategy has been defined to include the decision as to which diseases to vaccinate against, as well as the vaccine type, vaccination method, the timing of vaccination and the use of revaccination. In this lecture most important aquatic vaccines such as Bacterins, Live attenuated vaccines, killed vaccine and DNA vaccines would be discussed for some important fish bacterial and viral diseases. Also, monovalent and polyvalent and route and strategy of administration fish vaccines would be talked. Meanwhile, economic aspects related to fish vaccination, potential costs and benefits could be discussed.
All treatments for fever are based on the belief that fits is the result of 41 degrees Celsius temperature and it damages cells of the brain and body. At the same time, there is no evidence-based tests or concrete diagnosing methods to the belief that fits and brain damage is the result of pyrexia . Necessary ingredients to destroy brain cells and fits cannot be seen in fever. In pyrexia or absence of fever, a fainted patient fell on the floor with unconscious state and destroy cells of the brain, and necessary ingredients to become conscious are the same. When disease increases essential blood circulation and energy level also decreases. The vertical height between the heart and brain is more than one foot. When the disease becomes severe, the ability to pump the blood to the brain decreases. As a result of this brain cells are damaged. so the patient might be paralyzed or may even die. In pyrexia or absence of fever, when blood flow to the brain decreases and fits are formed. There is no other way than this to increase blood circulation to the brain. It is a sensible and discreet action of the brain to protect the life or organ. Recovery from Fits. The patient becomes conscious before the time to get decreasing the temperature of fever. When the fainted patient lie on the floor, the vertical height between the heart and brain is decreased, blood circulation increased to the brain. Self-checking methods. When the fainted patient lie on the floor, The patient can stand straight and lie on bed alternatively. Then the patient can experience himself the intensity of blood circulation. The patient can experience when he stands his blood circulation decreases and when lying on the bed the blood circulation decreases. Besides that, he can also experience increased blood circulation when lie on the bed raise the foot higher than the head.
First of all I would like to discuss about the global pandemic covid 19 outbreak across the Whole World & how much impact on human after spreading this virus globally.Most of the peoples has been said its (virus) originated from china ,Wuhan state.whatever this virus was increasingly day by day then time. Million of peoples were died by this killer virus across the whole world as well till present time.From that time many scientist were worried about this outbreak ,they Search new innovation how to get rid of the whole world .At last they partially successful to invention new vaccine .Drug regularity authority has approved emergency Authorization to give on human clinical trial like pfizer &moderna vaccine is found highly efficacy rate until now other than various vaccine. We are also inoculated this. At last we pray to almighty Allah to get rid of each of the part of the world free from this killer virus.
Background: Vaccine coverage is lower for influenza than for other vaccines, varying by age, race/ethnicity, and region. Vaccine safety concerns are common despite a lack of epidemiological evidence. We characterized vaccine hesitancy and identified associations with influenza vaccination.
Methods: Respondents ≥18 years old were recruited from a non-probability-based Internet panel survey (N=1,925). We measured sociodemographic characteristics, vaccine confidence, influenza vaccination history (2018-2019 and 2019-2020), trust in pharmaceutical companies and public health authorities, and perceived vaccine reaction history. The association between variables hypothesized to be associated with vaccine hesitancy and influenza vaccination was estimated with Taylor-linearized variance estimation for tabulations and Poisson regression for unadjusted and adjusted prevalence ratios. Backwards stepwise regression identified parsimonious, adjusted models (p<0.05).
Results: The weighted study population was 50.6% female, 61.8% White, non-Hispanic. 62.9% had a child <18 years old, and 47.1% had a high school education or less. High vaccine hesitancy was greatest among parents of young children (45.4% vs. 27.6% parents of teenagers vs. 37.7% adults without minor children). Awareness of federal vaccine safety oversight was low across age groups. In adjusted models of all age groups, higher education and use of complementary/alternative medicine (CAM) were associated with higher vaccination prevalence. Vaccination prevalence was lower among those with high vaccine hesitancy.
Discussion: We identified common vaccine misconceptions associated with vaccine hesitancy. CAM use and higher education were associated with vaccination across age groups. Vaccine hesitancy differed by parental age, which may have influenced results. Results are subject to selection and social desirability biases, though quotas were used to enroll a sample representative of sociodemographic distribution of the U.S.
Conclusions: Age and education-level appropriate, targeted communications are needed. Future research should investigate how to reach sociodemographic minorities, less likely to use CAM or be vaccinated, and whether raising public awareness of federal vaccine safety oversight improves confidence.