Sars-Cov-2 Vaccine from Infected Plant Attenuated Virus Using Acer Cincinatum (Vine Maple)

Purpose of study Proposal for an oral (or if required, parenteral) COVID-19 vaccination based upon this described technology. Investigational theory under study for the past 9 months of COVID-19 growing season. Coronavirus can attack and infect plant species. It was found that SARS-CoV-2 can infect various plant species. Others have found plants, for example tobacco as a good growth medium for Coronavirus and SARS-CoV-2. This current study has found various plants species infected with SARS-CoV-2 by rPCR. As the plants were located beside a well used hiking trail for humans, and were infected along the trail including various species with SARS-CoV-2, hypothesized that human airborne contact had caused infection in the bordering plants. Humans were observed to be coughing while walking on the trail, and were not wearing masks. The plant leaves developed small circular colonies of the virus, which became self-limited at several millimeters in diameter. All of the plants were clear of these lesions before the COVID-19 Pandemic. The plants 'immune' system produced antiviral agents, including lectins which limited the growth of the colonies and prevent death of the leaf and whole plant. The fungal cultures of the 'spots' were negative. The rPCR of all spots tested in the present series was positive for SARS-CoV-2. Hypothesis, that self-augmentation of the virus occurred by the natural culturing in plant leaves that produce antiviral agents as part of their 'immune system.' Hypothesis, a symbiotic type relationship developed between the plant using its chemical immune system, and the virus allowed to replicate in an augmented fashion to allow both the virus and the host to survive and grow. As the top candidates for the oral vaccine are nontoxic, hypothesis involves the maceration of the infected leaves, mixing with a nontoxic adjuvant and flavoring to promote assimilation and palatability, with the proposed route of entry being mastication, thus exposing the oral-nasal mucosa to the vaccine, with the probable best of immunity to usual exposure to the SARS-CoV-2 virus, that is the oral-nasal mucosal and upper airway route. As many types of animals are now infected with SARS-CoV-2, it is further hypothesized that this oral vaccine could also be mass produced to add to various animals by feedstock and oral route. Methods used Hypotheses formed through observations. Testing of observations by pPCR, viral cell culture, fungal culture, light and electron microscopy. Summary of results pPCR SARS-CoV-2 positive, cell culture 'lysis experiment' positive, EM and light microscopy positive, fungal culture negative. Conclusions TABLE OF HYPOTHESES AND STUDY RESULTS (HYPOTHETICAL, OBSERVED, PROVEN) 1. The first hypothesis that the virus is attenuated by the plant, using its innate chemical immune system. Similarly, Pasteur used chemical such as phenol to attenuate viruses for wome of the first successful vaccines. Observed. 2. Hypothesis, the plants 'immune' system produced antiviral agents, including lectins, flavonoids, and others, which limited the growth of the colonies and prevent death of the leaf and whole plant. Proven. 3. Hypothesis is that the nontoxic plants, such as Vine Maple sp.(Acer cincinatum), could be used to produce and oral plant attenuated vaccine. Hypothesis. 4. Hypothesis involves the maceration of the infected leaves, mixing with a nontoxic adjuvant and flavoring to promote assimilation and palatability, with the proposed route of entry being mastication, thus exposing the oral-nasal mucosa to the vaccine, with the probable best of immunity to usual exposure to the SARS-CoV-2 virus, that is the oral-nasal mucosa, upper airway. (Figure Presented).

CD8+ T CELL CLONES FROM PRIOR SARS-CoV-2 INFECTION PREDOMINATE AFTER mRNA VACCINATION

Background: Hybrid immunity is more protective than vaccination or prior infection alone. To understand the formation of hybrid immunity, we studied how SARS-CoV-2 mRNA vaccines interact with T cell memory by tracking spike (S) specific T cells in cohorts of hospitalized (n = 19) or non-hospitalized (n = 34) COVID-19 convalescents. We hypothesized that S-reactive CD4 and CD8 T cells would increase in response to serial vaccine doses and reflect prior immune exposure at the clonal level. Method(s): After vaccination, we stimulated PBMCs from 12 participants (8M/4F) with peptides spanning S. Activated cells (CD69+CD137+) were sorted and CD4/CD8 phenotype linked with paired TRB-TRA sequences at single cell resolution. S-reactive TRB sequences were mapped within 4-6 serial blood and post-booster nasal TRB repertoires to evaluate S-reactive CD4 and CD8 T cell clonotypic kinetics spanning convalescence to boost. PBMCs from 53 participants were sequenced with the ImmunoSEQ assay to evaluate S-reactive TRB breadth using a database of S-assigned TRB sequences (Adaptive Biotechnologies), comparing S-reactive TRB diagnostic breadth by hospitalization status (Wilcoxon test). Result(s): SARS-CoV-2 mRNA vaccination provoked strong T cell clonal expansion in most participants. At 8-12 months after infection, each primary mRNA dose increased the abundance and diversity of S-specific T cells. Clonal and integrated expansions were larger in CD8 than in CD4 T cells. At the convalescent time point, we observed greater diagnostic S-reactive CD4 T cell breadth in hospitalized vs. non-hospitalized patients (p< 0.01). CD4 T cell S breadth was again higher in previously hospitalized persons after the 2nd primary (p=0.02) and booster (p< 0.01) doses, suggesting that diverse CD4 T cell memory after severe infection leads to increased repertoire diversity after vaccination. S-specific T cells with identical TCRs were detectable in blood and the nasal mucosa, with specificity confirmed using a TRA/TRB transgenic T cell with the matching receptor. Conclusion(s): Although both S-specific CD8 and CD4 T cell memory are established by prior infection, S-specific CD8 T cells predominated in blood after primary vaccination, with some clonotypes showing up to 1000-fold expansion across 1-2 mRNA doses. Vaccine-reactive CD8 clonotypes were present at the barrier nasal site after booster mRNA dosing. Severe disease imprinted a highly diverse S-reactive CD4 repertoire persisting through vaccination.

Seroconversion in nasal lining fluid antibodies after Covid19 infection in a health professional cohort

Background: The novel coronavirus SARS-CoV- 2 has caused far-reaching consequences world-wide. Lack of immunity in human, severe airway disease based on a high virulence and its airborne transmission pointed to a significant role of the airways. To investigate immune responses and antibody seroconversion in nasal lining fluid and in serum we examined a cohort of health professionals at the university hospital Klinikum rechts der Isar in Munich. By long-term follow-up of infected and non-infected participants, we were able to investigate the development of local and systemic immunity against SARS-CoV- 2. Method(s): To learn about nasal antibody production we reached out for hospital staff with estimated high risk of a possible Covid19 infection due to their working conditions and staff members suffering from symptoms like fever, cough, loss of smell and taste or sore throat. To detect current infections, we performed SARS-CoV- 2 PCR testing at visit one (V1) and asked our participants to rate possible Covid19 symptoms by filling a questionnaire before every sampling. We eventually included participants, who had been tested positive for SARS-CoV- 2 before (n = 22), as well as people without detected infection, including high-risk contact persons of Covid19 patients and individuals with no Covid-19 infection so far (n = 85). The cohort included 107 hospital staff members, who were sampled six times overall between March and September 2020. Each of the six visits V1 -V6 contained the sampling of serum and nasal fluid to measure IgG, IgM, and IgA rates using immunoassay technique. Result(s): We were able to show the increase of IgA and IgG in the nasal mucosa after recent Covid19 infection. In infected individuals the levels of SARS-CoV- 2 specific nasal IgA increased until V2 with a mean of 4,81 +/- 1,92 mug/l compared to a mean of 0,13 mug/l in non-infected participants, followed by a plateau until V4 and decreased again until V6. Nasal IgG showed a similar trend, apart from a steeper decline after reaching a peak on V2 with a mean of 7,39 +/- 1,63 mug/l, which correlated to the antibody responses in serum. Non-infected individuals showed a mean level of 0,03 mug/l nasal IgG on V2. Serum IgA declined from V1 onwards and hereby showed a quicker drop of systemic antibody levels compared to the nasal lining fluid. Nasal antibody rates reached peaks of 40,00 mug/l (nasal IgA) and 25,74 mug/l (nasal IgG). However, these counts will need further confirmation by a vaccinated control group. Conclusion(s): Nasal measurement of SARS-CoV- 2 specific antibodies provides deeper understanding of mucosal processes while facing inflammation, which may pave the way to less invasive diagnostic possibilities in the future. Furthermore, nasal antibodies built-up after an infection with Covid19 may be protective features concerning a possible re-infection with the virus.

Diagnosis of Nasal Breathing Using a Rhinomanometer in Patients with Osa Prescribed Conservative Treatment with Intraoral Devices

In recent years, diagnostics in the field of medicine has developed at an extremely rapid pace, thanks to the use and improvement of new medical devices and devices. The problem of timely and adequate diagnosis and treatment of the syndrome of obstructive sleep apnea (OSA) and snoring is particularly relevant world-wide. The disease has been proven to worsen the patients' quality of life, and may even threaten it. Obstructive sleep apnea and snoring syndrome (OSAS) is a widespread disease of social importance in which there is a reduction or cessation of airf low through the nose/mouth during sleep due to upper airway collapse. Obstructive sleep apnea affects the cardiovascular, endocrine, neurocognitive and other systems of the body. There are symptoms of loud snoring, choking, hypoxemia, and micro-awakenings, leading to sleep frag-mentation, daytime fatigue, and sleepiness. The latter greatly worsens the quality of life of patients. There are real risks to the life and health of the patient and others, given the possibility of falling asleep at the wheel in drivers with sleep apnea and participation in traffic accidents. To diagnose the syndrome, a poly-somnographic study is performed, which is still the gold standard. For a better diagnosis, it is recommend-ed to combine it with rhinomanometry. Treatment of OSA includes control of risk factors and removal of obstructive factors that make breathing difficult. Severe OSA syndrome is treated with continuous positive pressure ventilation (CPAP) during sleep, possibly in combination with intraoral devices. Rhinomanome-try can also be used to monitor the effectiveness of CPAP therapy in severe forms of the syndrome by deter-mining tissue resistance. The impact on patients with a milder form of OSA treated with intraoral devices is also monitored. The method can also be used in patients with allergic rhinitis, sinusitis of rhinogenic and other origin, and patients with orthodontic deformities. The correct choice of intraoral appliances for conservative treatment of OSA and timely diagnosis are key to successful treatment.Copyright © 2022, Bulgarian-American Center. All rights reserved.

Post-COVID anosmia and the expression of pathological proteins in nasal mucosa nerves endings

Introduction: Anosmia and ageusia were observed as frequent neurological complications of SARS-CoV-2 infections. the aim of the study was to elaborate methods for detection of pathological proteins in nerve endings and to evaluate the frequency and intensity of pathological proteins expression in patients with persistent anosmia. Material(s) and Method(s): the study included 249 patients (181 females and 68 males) aged 47 +/-14 years from NeuroCOViD Polyclinic in Poznan observed from April 2021 untill now. the mucosal biopsy was performed using endoscopy from anterior ethmoid cells. the expression of alpha-synuclein was evaluated using immunofuorescence, and amyloid, tau and tDP43 proteins-using immunohistochemistry. Result(s): Anosmia was observed in 42% of patients and cacosmia-in 6%. Ageusia/dysgeusia was observed in 31% cases. in patients with mild clinical course of COViD19-not hospitalized anosmia (45%) and dys-geusia were more frequent (33%), and cacosmia was observed only in this group. in hospitalized patients anosmia was found in 22% of cases, dysgeusia in 13%, and cacosmia was not observed at all. the expression of alpha-synuclein, amyloid, tau and tDP43 proteins was found in nerve bundles, epithelial cells and in surrounding (nerve endings) of gland cells. Conclusion(s): SARS-CoV-2 infection may induce the expression of pathological proteins in olfactory mucosa of post-COViD patients with anosmia.

Spotting the Needle Amongst the Haystack - When Asthma Isn't Asthma

Introduction/Background: Eosinophilic granulomatosis with polyangiitis (EGPA), previously known as Churg-Strauss Syndrome, is a rare, small to medium vessel ANCA associated vasculitis. Hallmarks of EGPA include asthma, chronic rhinosinusitis, and peripheral neuropathy. EGPA is characterized by a prodrome of asthma and allergic rhinitis, followed by peripheral blood hyper-eosinophilia and accumulation of extravascular eosinophils, and finally systemic vasculitis. Extrapulmonary involvement is common, sometimes with fatal outcomes. The onset of EPGA is typically between 25-50 years;however, EGPA also occurs during childhood and has a significant morbidity and mortality. Description/Method: Our patient presented to the emergency department with a 2-week history of lethargy, wheeze and left sided neck swelling. After testing COVID-19 positive eight months prior to this, she developed wheezy episodes and was subsequently diagnosed with asthma which was managed with bronchodilators as required. She was reviewed by an allergist who confirmed a dust mite allergy and prescribed Montelukast. She remained well during the summer months however during winter she had 3 distinctive episodes of wheeze and cough which were managed by antibiotics and prednisolone. In the emergency department, an echocardiogram was performed which showed a cardiac tamponade. She was transferred to CICU where she had a pericardial drain inserted. The fluid was abundant with inflammatory cells. Multiple investigations were performed as follows: Hb: 135g/L, wbc: 20.30 x 10 9/L, Eosinophils: 12.77 x 10 9/L, CRP: 51 mg/L, ESR: 75 mm/hr, LDH: 1188 IU/L, IgE: 8000 UI/ml, ANA, ANCA: negative. CT chest showed mediastinal lymphadenopathy and patchy bilateral infiltrate and cardiac MRI showed myopericarditis and LV fibrosis. BMA showed no malignant cells and sinusitis was confirmed by CT. On examination, she was underweight. Her nasal mucosa looked inflamed. Otherwise systemic examination was unremarkable. In the context of poor ejection fraction (20%) with LV fibrosis, urgent MDT was arranged and concluded that our working diagnosis was EGPA. The decision was made to start IV methylprednisolone 10mg/kg/day for 3 days and Ivermectin. That night our patient had a VF arrest which required a single shock conversion 4J/kg. There was 7-minute downtime. Treatment was escalated to include cyclophosphamide, rituximab and plasmapheresis. The patient made a remarkable recovery, extubated and transferred to a normal ward. Her eosinophils count and inflammatory markers improved dramatically following treatment. However, she developed severe neuropathic left leg pain and NCS confirmed peripheral neuropathy Discussion/Results: EGPA is a very rare disease and diagnosis can be challenging especially with the absence of histopathology diagnosis. Early empirical treatment especially in a very ill child in intensive care unit can save lives and divert the progress of the disease. This patient has fulfilled the American College of Rheumatology criteria to diagnose EGPA including asthma, eosinophil count > 10% of upper normal, peripheral neuropathy, pulmonary infiltrates on CT thorax and paranasal sinuses abnormalities. Cardiac biopsy of the fibrotic mass may be a useful tool for diagnosis;however, this invasive procedure may expose this patient with high risk of fatal arrhythmias. Since other causes of eosinophilia were ruled out including parasitic infections, lymphoproliferative disorders, and rare primary immunodeficiency syndromes (hyper-IgE syndrome due to STAT3 or DOCK8 deficiency and Omenn syndrome) and the patient responded well to treatment, the diagnosis of EGPA was supported. Key learning points/Conclusion: Asthma not responding to bronchodilator could be another diagnosis Eosinophilia should be interpreted with caution. Defer the need for histopathology diagnosis in critically ill children Cardiac involvement is a life-threatening marker Early diagnosis prevents life threatening complications.

Study on the Mucosal and Serological Immune Response to the Novel Coronavirus(SARS-CoV- 2) Vaccines

Vaccines that elicit mucosal immune responses against SARS-CoV-2 could potent ially be of exceptional importance in providing first line defence at the site of viral entry. In order to understand the mucosal immune response profiles of SARS-CoV-2 vaccines, we examined both the mucosal and systemic responses of subjects vaccinated by two different vaccination platforms: mRNA (Comirnaty) and inactivated virus (CoronaVac). Nasal epithelial lining fluid (NELF) and peripheral blood samples were collected in subjects who had received two doses of CoronaVac or Comirnaty. We quantified IgA and IgG specific to SARS-CoV-2 S1 protein, neutralisation antibody by ELISA in NELF and plasma samples. Only Comirnaty induced nasal S1-specific IgA and IgG responses, which were evident as early as on 14±2 days after the first dose. The NELF samples of 72% of subjects became IgA+IgG+, while in 62.5% of subjects the samples were neutralising by 7±2 days after the second dose. In 45% of the subjects their NELF remained neutralising 50 days after the booster. In plasma, 91% and 100% Comirnaty subjects possessed S1-specific IgA+IgG+ on 14±2 days after the first dose and 7±2 days after booster, respectively. The plasma collected on 7±2 days after booster was 100% neutralising. The induction of S1-specific antibody by CoronaVac was IgG dominant, and 70% of the subjects possessed specific IgG by 7±2 days after booster and were all neutralising. This study reveals that Comirnaty is able to induce S1-specific IgA and IgG r esponse with neutralising activity in the nasal mucosa in addition to a consistent systemic response.

COVID-19 associated mixed mold infection: a rare case report of aspergillosis and mucormycosis

Background: Fungal epidemic was announced amid COVID pandemic with several cases of COVID-associated Mucormycosis and Aspergillosis being reported however there is not sufficient data regarding mixed fungal infection. Case Study: A 54-year-old male patient diagnosed with severe COVID 19 pneumonia and diabetes 1-month back presented to OPD with C/O of chest pain and breathlessness for two days associated with haemoptysis, heaviness and congestion of right nostril but no fever. O/E patient was tachypnoeic, hypoxic and in shock, Neutrophil count 87%, RBS-530 mg/dl, urine ketone body was absent. Chest x-ray showed opacity over the left upper and mid-zone, HRCTthorax showed a bird-nest-sign noted in the left upper lobe S/O invasive fungal infection. MRI PNS showed mucosal thickening S/O sinusitis, Fungal infection. Sino-nasal mucosa KHO-mount and fungal culture showed mixed infection of Rhizopus species and aspergillus flavus. Right nasal HP study showed mixed invasive moulds infection. Initially, the patient was treated conservatively later on inj. amphotericin-B was started. The patient's condition worsened on day-18 and succumbed a day later. Discussion: Uncontrolled diabetes-mellitus, and corticosteroids leading to hyperglycaemia, extensive use of broad-spectrum antibiotics increases the risk of invasive Moulds. In our case study, patients suffered from COVIDpneumonitis and had uncontrolled diabetes leading to damage of airway epithelium inviting an invasion of tissues by moulds. Conclusion: Mixed fungal infections as COVID-19 sequelae may be an emerging issue and seen particularly in post- COVID patients with uncontrolled diabetes, and on steroids. The focus should be on prompt management: hit hard approach with both medical and surgical treatment.

Evaluation of il-6 levels in covid-19 patients whit smell disorders

Smell dysfunction is one of the most frequent symptoms in COVID-19 patients. In the early stages of the disease it allows to identify positive subjects. The odorous substances recognize two different systems in the olfactory epithelium: the olfactory and the trigeminal systems that coexist and interact in the processing of sensory information. In COVID-19 patients there is an inflammatory reaction of the nasal mucosa. Infected supporting cells of the nasal mucosa release molecules that activate the local antiviral innate immune response. In fact, macrophages spread inflammatory mediators, in particular TNF-η , IL-6 and IL-1. In this study we compared IL-6 levels with the degree of olfactory disorders and with the type of unperceived odour.Materials and methodsFrom 15 March to 30 November 2020 have been selected 82 patients (45 men age 62.3 ±14.2 and 37 women age 57.1± 12.8) with only smell dysfunctions were divided into mild and moderate patients. The evaluation of the smell disorder was carried out with a 14 questionnaire relating to the perception of domestic odorous: 6 questions for olfactory sensitivity (own perfume usually sprayed, oregano, olive oil, nutella, coffee aroma, orange juice) and 8 for olfactory-trigeminal sensitivity (alcohol, fish odor, vinegar, mint (gum), toothpaste, shampoo, cheese, ammonia).The IL-6 (v.n. 0 - 7 pg/ml) was measured with chemiluminescence assay using Cobas e801 (Roche Instrumentation). Statistical analyses were performed with Wilcoxon Rank test, and Mann-Whitney test (p <0.05). ResultsThe trigeminal and olfactory sensitivity are more compromised in moderate than mild patients (p <0.05). The statistically significant differences there were in IL6 levels in moderate versus mild patients when there was an impairment of trigeminal sensitivity (p <0.05). Conclusion In this study suggested that the smell disorders in Covid-19 patients couldn't be a deficit of the olfactory central nervous pathways but could be rather than mainly associated with the inflammatory process of the nasal mucosa and that deficit of the type of domestic unperceived odour ('olfactory' or 'trigeminal' sensitivity) could indicate the degree of severity of the disease.

THE COVID-19 EFFECTS ON DENTISTRY

The rapid spread of the COVID-19virus and related pneumonia has posed a major challenge for healthcare systems worldwide. The infection was discovered in the city of Wuhan, Central of China and swept across the world.Theincubation period of the viruscan range from 1-14 days, and the virus spread can happen in the absence of clinical symptoms as well.The most frequently reported symptoms are fever, cough, dyspnea, and myalgia or fatigue. Modes for transmission of virus include direct transmission through coughing,sneezing, and inhalation of droplets and contact transmission through contact with nasal, oral, and ocular mucosa. Droplet and aerosol transmission of the virus are the most common causes of COVID-19 infection in dental clinics and hospitals.COVID-19 virus has lately been detected in saliva of infected patients, thus posing an alert to health professionals to be customarily vigilant in protecting against the infectious disease spread. Because of the presence of virus in saliva, it may be helpful as a non-invasive tool in the rapid detection of the virus.During this pandemic dissemination of COVID-19, dental treatment must be confined to the procedures that cannot be deferred. All the precautions must be taken in terms of triaging, personal protective equipment, hand hygiene, pre-procedural mouthrinse, use of rubber dam, disinfection of the surfaces. Dental professionals are at the highest risk of COVID-19 infection;hence, dental practicehas to be reorganized in order to ensure higher safety standards for both dentists and patients.

Black Fungus: A Lethal Communal Issue After Winning the Life Battle Against COVID-19

The authors aimed to give a quick reference guide for humanity about the new threat even after the COVID-19 health battle i.e., the Black fungus also baptized as Mucormycosis infection. After recovering from COVID-19, patients with diabetes mellitus and patients who have undergone steroidal treatment are more prone to black fungus infection. This review gives quick information about various types of Mucormycosis infections, risk factors, symptoms, treatment, and prevention of black fungus. The things that can be and cannot be done to eradicate the black fungus. Any sign of black nasal mucosa/sputum, fever, headache, hazy/blurred/double vision with eye pain, loss of one side sensation on face and loss in the sensation while chewing, etc., then it should not be neglected and immediately intimated to the health professionals and get treated. If black fungus is unidentified early stages and untreated in time, the patients may lose their facial/neck parts as a part of cleaning surgery. The study concludes that by maintaining hygienic conditions, health checkups, and doctors' advice one can fight and eradicate the black fungus.

Efficacy of recombinant interferon α-2b in the complex therapy of covid-19

Objective. To evaluate the efficacy and safety of VIFERON® (recombinant interferon α-2b) rectal suppositories and gel for external and local use in the complex therapy of COVID-19 in adult patients. Patients and methods. 140 patients with COVID-19 were included in a prospective comparative controlled research study. The main group consisted of 71 patients who received standard therapy for coronavirus infection in combination with VIFERON® (rectal suppositories 3.000.000 IU for 1 supp. 3 times a day and gel 36.000 IU/g 5 times a day on the surface of nasal mucosa and palatine tonsils for 14 days);the comparison group – 69 patients who received standard therapy. Results. Patients who received VIFERON® complex therapy demonstrated more rapid clinical improvement by reducing the duration of symptoms of weakness and intoxication compared to the patients who received only standard therapy. The high activity of VIFERON® in elimination rate of SARS-CoV-2 RNA in nasopharyngeal samples was shown. The use of VIFERON® made it possible to achieve a stable concentration of IL-6 in the blood serum during the entire follow-up period. There were no adverse events associated with the administration of VIFERON®. Conclusion. The results obtained indicate the superiority of therapy with the inclusion of VIFERON® over standard therapy. Given the favorable safety profile, the studied regimen can be recommended for the treatment of adult patients, especially from risk groups, where the possibilities of conventional etiotropic therapy are limited due to possible toxicological effects.