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Objective: COVID-19 has emerged as a significant global health crisis causing devastating effects on world population accounting for over 6 million deaths worldwide. Although acute RTI is the prevalent cause of morbidity, kidney outcomes centered on a spectrum of AKI have evolved over the course of the pandemic. Especially the emerging variants have posed a daunting challenge to the scientific communities, prompting an urging requirement for global contributions in understanding the viral dynamics. In addition to canonical genes, several subgroup- specific accessory genes are located between the S and E genes of coronaviruses regarding which little is known. Previous studies have shown that accessory proteins (aps) in viruses function as viroporins that regulate viral infection, propagation and egress [1]. In this study we attempted to characterize the function of aps of coronavirus variants as ion channels. Furthermore, we also probed the interaction of ap4 with the host system. Method(s): Serial passaging (selection pressure), growth kinetics, confocal imaging, genome sequence analysis and proteomics were performed in Huh-7, MRC5 cells and/or human monocyte derived macrophages. Potassium uptake assay was performed in a Saccharo myces cerevisiae strain, which lacks the potassium transporters trk1 and trk2. Ion conductivity experiments were performed in Xenopus laevis oocytes using Two Electrode Voltage Clamp (TEVC) method. Result(s): Serial passaging demonstrated the acquisition of several frameshift mutations in ORF4 resulting in C-terminally truncated protein versions (ap4 and ap4a) and indicate a strong selection pressure against retaining a complete ORF4 in vitro. Growth kinetics in primary cells illustrated a reduction of viral titers when the full-length ap4 was expressed compared to the C-terminally truncated protein ap4a. Confocal imaging showed that ap4 and ap4a are not exclusively located in a single cellular compartment. Potassium uptake assay in yeast and TEVC analyses in Xenopus oocytes showed that ap4 and ap4a act as a weak K+ selective ion channel. In addition, accessory proteins of other virus variants also elicited microampere range of currents. Conclusion(s): Our study provides the first evidence that ap4 and other accessory proteins of coronavirus variants act as viroporins. Future studies are aimed at demonstrating the role of ap4 during the viral life cycle by modulating ion homeostasis of host cell in vivo (interacting proteins obtained from proteomic studies) and thereby serve as a tool for potential drug target.
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Vaccines that are delivered orally have several advantages over their counterparts that are administered via injection. Despite the advantages of oral delivery, however, approved oral vaccines are currently limited either to diseases that affect the gastrointestinal tract or to pathogens that have a crucial life cycle stage in the gut. Moreover, all of the approved oral vaccines for these diseases involve live-attenuated or inactivated pathogens. This mini-review summarizes the potential and challenges of yeast oral vaccine delivery systems for animal and human infectious diseases. These delivery systems utilize whole yeast recombinant cells that are consumed orally to transport candidate antigens to the immune system of the gut. This review begins with a discussion of the challenges associated with oral administration of vaccines and the distinct benefits offered by whole yeast delivery systems over other delivery systems. It then surveys the emerging yeast oral vaccines that have been developed over the past decade to combat animal and human diseases. In recent years, several candidate vaccines have emerged that can elicit the necessary immune response to provide significant protection against challenge by pathogen. They serve as proof of principle to show that yeast oral vaccines hold much promise.
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Introduction: Disseminated histoplasmosis (DH) presents as primarily lung manifestations with extrapulmonary involvement in immunocompromised hosts. Granulomatous hepatitis as first presentation of DH in an immunocompetent host is uncommon. Case Description/Methods: 25-year-old female presented with one month of fever, fatigue, myalgias, 30-pound weight loss, cough, nausea, vomiting, and epigastric pain. She has lived in the Midwest and southwestern US. Presenting labs: TB 1.9 mg/dL, AP 161 U/L, AST 172 U/L, ALT 463 U/L. Workup was negative for COVID, viral/autoimmune hepatitis, sarcoidosis, tuberculosis, and HIV. CT scan showed suspected gallstones and 9 mm left lower lobe noncalcified nodule. EUS showed a normal common bile duct, gallbladder sludge and enlarged porta hepatis lymph nodes which underwent fine needle aspiration (FNA). She was diagnosed with biliary colic and underwent cholecystectomy, with white plaques noted on the liver surface (A). Liver biopsy/FNA showed necrotizing granulomas (B) and fungal yeast on GMS stain (C). Although histoplasmosis urine and blood antigens were negative, histoplasmosis complement fixation was >1:256. She could not tolerate itraconazole for DH, requiring amphotericin B. She then transitioned to voriconazole, discontinued after 5 weeks due to increasing AP. However, her symptoms resolved with normal transaminases. At one year follow up, she is asymptomatic with normal liver function tests. Discussion(s): DH is a systemic granulomatous disease caused by Histoplasma capsulatum endemic to Ohio, Mississippi River Valley, and southeastern US. DH more commonly affects immunocompromised hosts with AIDS, immunosuppressants, and organ transplant. Gastrointestinal involvement is common in DH (70-90%) with liver involvement in 90%. However, granulomatous hepatitis as primary manifestation of DH is rare (4% of liver biopsies). Hepatic granulomas are seen in < 20%. Patients may present with nonspecific systemic symptoms. Serum/urine antigens may be negative. Gold standard for diagnosis is identifying yeast on tissue stains. Recommended treatment is amphotericin B followed by 1 year of itraconazole. However, shorter treatment duration may be effective in immunocompetent hosts. This case is unique in that granulomatous hepatitis was the first presentation of DH in our immunocompetent patient diagnosed on EUS FNA and liver biopsy. Clinicians must have a high degree of suspicion for DH in patients with fever of unknown origin especially in endemic areas regardless of immunologic status. (Table Presented).
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Objective To determine the risk factors for developing secondary fungal pneumonia in moderate to severe coronavirus disease 2019 (COVID-19) cases. Using predictors of fungal infection helps to guide the diagnosis and treatment in these cases and save their lives. Patients and methods A total of 257 patients with moderate to severe COVID-19 pneumonia were examined in this retrospective study at Al Qassimi Hospital of EHS. An assessment of clinical, laboratory, and radiologic findings was performed upon admission. The data were collected and analyzed. Results Overall, 32% of critically ill COVID cases had fungal infection;47% of them were candida, whereas aspergillosis and yeast were positive in 26.5% each. At the time of hospitalization, computed tomography chest findings had a strong correlation with fungal culture results in COVID-19 cases. Fungal infection in COVID-19 cases correlated strongly with metabolic acidosis, high erythrocyte sedimentation rate, high blood sugar, need for mechanical ventilation at admission, vasopressor use, renal replacement, long duration of steroid treatment, long stay in ICU, and long duration on mechanical ventilation. The longer the duration of PCR positivity, the higher the incidence of positive sputum fungal culture result. Conclusion COVID-19-infected patients with other risk factors for fungal infections should always be considered to have fungal infections if pathogenic organisms are isolated from respiratory secretions or other microbiological or immunological markers appear positive. Computed tomography chest finding in COVID-19 cases is an important predictor for fungal infection.Copyright © 2023 The Egyptian Journal of Chest Diseases and Tuberculosis.
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Background: The continued emergence of severe acute respiratory syndrome coronaviruses (SARS-CoVs) and recent explosion of the SARS-CoV-2 pandemic highlights the need for broad and potent antibody recognition and understanding the contexts in which they may develop. Antibodies with cross reactivity across SARS lineages may be of particular value in preparing for future outbreaks of new sarbecoviruses. Method(s): We isolated monoclonal antibodies (mAbs) from an individual 60-days post-vaccination, 30-days post Delta-infection. Reconstructed antibodies were screened for binding to a panel of prefusion-stabilized Spike trimers from SARS-CoV-2 and other beta-coronaviruses using enzyme-linked immunosorbent assay (ELISA). Neutralization potency and breadth was assessed using a spike-pseudotyped lentivirus neutralization assay. Additionally, epitope and escape mutant profiling was conducted by deep mutational scanning (DMS) to identify mutations that affect antibody binding. Lastly, binding breadth was further evaluated using a yeast display library of RBDs from SARS-CoV-2 variants and related sarbecoviruses. Result(s): We identified several SARS-CoV-2-specific mAbs that neutralized SARS-CoV-2 variants of concern (VOCs) and SARS-CoV-1. Notably, two of these mAbs (C68.61 and C68.185) neutralized SARS-CoV-1 with an IC50 = 307 and 139 ng/mL (respectively) that is similar to or better than the potency of S309 (IC50 = 206 ng/mL) and CR3022 (IC50 = 981 ng/mL), which are mAbs isolated from individuals with SARS-CoV-1 infections. C68.61 also neutralized all Omicron VOCs tested and retained neutralization activity against currently circulating variants BQ1.1 (IC50=790 ng/ml) and XBB (IC50=590 ng/ml). Key C68.61 mAbescape mutations identified by DMS in the Omicron BA.2 background yeast display library included sites K462, E465, R466, and I468, which are conserved sites across all VOCs and SARS-CoV-1. The isolated mAbs displayed crossreactive binding to RBDs from diverse SARS-CoV-1-related CoVs and African and European sarbecovirus isolates as well as SARS-CoV-2 VOCs. Conclusion(s): Here we describe mAbs from a SARS-CoV-2-infected individual that bound and neutralized both SARS-CoV-2 and SARS-CoV-1, including one that showed breadth across recent VOCs. Given their breadth, these SARS-CoV-2 cross-reactive mAbs may be robust to viral escape and thus could contribute to therapeutic efforts. In addition, these mAbs displayed broad cross-reactive activity across sarbecoviruses and may be beneficial against future spillover events.
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Case Report: Cryptococcosis is an opportunistic infection caused by the encapsulated yeast Cryptococcus, with C. neoformans and C. gattii being the most common species to cause human disease. Immunocompromised individuals are predisposed to infections with C. neoformans, which has known predilection to CNS and pulmonary lymph nodes. We present a unique case of disseminated cryptococcosis in the setting of end-stage renal disease (ESRD), cirrhosis, tumor necrosis factor inhibitor use and steroid use for COVID19. Method(s): A single-patient case report was conducted after IRB approval. Case Presentation: A 55-year-old woman with uncontrolled diabetes, lupus, rheumatoid arthritis on adalimumab, hepatitis C status post boceprevir, cirrhosis, former IV drug use, and ESRD on hemodialysis via bovine arterial-venous fistula graft presented with worsening dyspnea, cough, and altered mental status. Three months prior, patient was admitted to an outside hospital for COVID19, complicated by pulmonary embolism status post anticoagulation therapy. Patient was treated with an unknown steroid regimen, which was continued by a second outside facility when symptoms failed to improve. Patient then presented to our facility 24 hours after discharge due to continued symptoms. On admission, patient was noted to have altered mentation and hypoxia with pulmonary edema on chest x-ray and was urgently hemodialyzed. Further work-up was obtained due to non-resolving symptoms, including blood and sputum cultures, cocci serology and QuantiFERON gold. CT chest revealed bilateral consolidations. Patient was started on antibiotics for presumed hospital-acquired pneumonia. During the hospital stay, preliminarily blood cultures grew yeast and patient was started on Micafungin. However, Micafungin was changed to Liposomal Amphotericin B as ovoid structures seen on gram stain could not confirm nor rule out cryptococcus. Subsequent bronchial wash and bronchoalveolar lavage cultures, as well as final blood cultures resulted Cryptococcus neoformans. Serum cryptococcus antigen returned reactive, titer 1:512. Antibiotics were discontinued and Isavuconazonium was started with Liposomal Amphotericin B. Due to recurrent headaches, lumbar puncture was obtained and revealed lymphocytic pleocytosis without cryptococcal antigenicity. Patient completed 14 days of Liposomal Amphotericin B and Isavuconazole with continuation of Isavuconazole upon discharge. Conclusion(s): Disseminated cryptococcosis in non-HIV patients is rare in the modern HIV era. Clinicians should be aware and include it in their differential of any patient with multiple risk factors for opportunistic infection. In patients with cirrhosis and ESRD, treatment is limited given altered pharmacokinetics. Studies have shown improved survival with the addition of Isavuconazole in patients with disseminated cryptococcosis with CNS involvement in the setting of chronic liver disease and ESRD.
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Background: Pathogen surveillance is crucial but has become more challenging in the era of highly effective modulator therapy (HEMT), with many people with cystic fibrosis (PwCF) noting a considerable reduction or even absence of sputum on elexacaftor/tezacaftor/ivacaftor (ELX/TEZ/IVA). This challenge has been exacerbated by reduced face-toface contact with patients because of variable COVID-19 government restrictions on travel and social interaction (lockdowns) since March 20, relying on PwCF returning high-quality samples by mail. For those with pre-established bronchiectasis, it is likely that chronic infections and risk of new acquisition of infections remain on ELX/TEZ/IVA, although registry data suggest less prevalence of respiratory microorganisms on IVA [1]. We aimed to examine the impact of ELX/TEZ/IVA on frequency of respiratory pathogen surveillance and microorganism growth in our large, adult CF center. Method(s): A retrospective analysis of pathology results from respiratory samples received from March 19 to December 21, 2020, was completed for all patients commenced on ELX/TEZ/IVA at our CF center. Result(s): Respiratory samples from 216 PwCF who had commenced ELX/ TEZ/IVAwere analyzed. Median start date of ELX/TEZ/IVAwas October 10,2020. Before ELX/TEZ/IVA, the average number of respiratory samples per month was 108. This declined by 55% to an average of 48 per month when the first 50 PwCF commenced on ELX/TEZ/IVA and to 20 per month (82% reduction from pre-ELX/TEZ/IVA) when 100 PwCF had commenced ELX/ TEZ/IVA. The number of positive samples per month decreased from January 20, 2020, correlating with the introduction of ELX/TEZ/IVA and the reduction of respiratory samples received (Figure 1). The proportion of cough swabs and sputum samples remained similar from March 19 to December 21, 2020. (Six-month average showed that 19% of samples were cough swabs and 80% sputum for March to June 2019 and July to December 2021). We found no significant changes in proportion of samples positive for non-Pseudomonas spp. gram-negative organisms, Burkholderia spp., or gram-positive organisms (predominantly S. aureus) isolated over the period. There was a reduction by more than 50% of fungi and Candida spp. and a slight trend toward an increase in Pseudomonas spp. (mainly P. aeruginosa). Forty percent of PwCF who had one respiratory sample after ELX/TEZ/IVA initiation and 20% of those who had two or more samples showed a change in organism growth after ELX/TEZ/IVA initiation. In nearly 50% of these cases, the organism changed from gram negative to gram positive (P. aeruginosa to S. aureus in 69% of cases).(Figure Presented) Figure 1. Number of positive respiratory samples per month and number of people with cystic fibrosis on elexacaftor/tezacaftor/ivacaftor March 19 to December 21, 2020 Conclusion(s): The introduction of ELX/TEZ/IVA and its impact on sputum production has reduced surveillance of our patients' respiratory microbiology. This has been exacerbated by reduced face-to-face contact with patients due to the COVID-19 pandemic. The trends showa reduction in the isolation of fungi and yeasts and a slight increase in isolating Pseudomonas spp. In those who we have seen a change in organism growth, many have gone from gram-negative to gram-positive organisms. These data highlight the challenges of monitoring for new positive cultures and changes in microbiology cultures in the era of HEMT.Copyright © 2022, European Cystic Fibrosis Society. All rights reserved
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The novel COVID-19 vaccines have been instrumental at transforming the pandemic into an endemic disease. However, many contemporary vaccines, especially the landmark mRNA vaccines, require cold storage that makes them difficult for low income and developing countries to keep and distribute, and no shelf stable, low-cost alternative currently exists. In response to this need, we are developing a novel COVID-19 vaccine delivery system using the probiotic yeast Saccharomyces boulardii. We engineered an integrating construct to express the receptor binding domain (RBD) of the SARS-CoV-2 spike protein tagged with the yeast pheromone secretion signal and with the Claudin-4 targeting sequence of the Clostridium perfringens enterotoxin. Preliminary data from two animal trials suggest that our candidate yeast oral COVID-19 vaccine can trigger a robust humoral immune response in mice. Experiments are underway to assess its effect on the murine T-cell response. Our laboratory is supported in part by a research grant from the PCHRD-DOST of the Republic of the Philippines.Copyright © 2023 The American Society for Biochemistry and Molecular Biology, Inc.
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This review is aimed to explore the health beneficial effects of probiotics which are live microorganisms that provide a positive health influence on humans when taken in sufficient quantity. Lactic acid bacteria, bifidobacteria, and yeast are frequently used as probiotics. These health-beneficial bacteria could compete with pathogens and modulate the gut microbiota, and exhibit immunomodulatory, anti-obesity, anti-diabetic, and anti-cancer activities which are discussed in this review. Moreover, recent studies showed that probiotics could neutralize COVID-19 infections. Hence, probiotics have become an alternative to several drugs including antibiotics. In addition, probiotic efficacy also depends on the delivery system as the delivery agents help the bacteria to survive in the harsh environment of the human gut. Considering these health benefits of probiotics, now it has been applied to different food materials which are designated as functional food. This review explored a portrait of the beneficial effects of probiotics on human health.Copyright © 2022 The Author(s)
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Mycoses are infectious diseases caused by fungi, which incidence has increased in recent decades due to the increasing number of immunocompromised patients and improved diagnostic tests. As eukaryotes, fungi share many similarities with human cells, making it difficult to design drugs without side effects. Commercially available drugs act on a limited number of targets and have been reported fungal resistance to commonly used antifungal drugs. Therefore, elucidating the pathogenesis of fungal infections, the fungal strategies to overcome the hostile environment of the host, and the action of antifungal drugs is essential for developing new therapeutic approaches and diagnostic tests. Large-scale transcriptional analyses using microarrays and RNA sequencing (RNA-seq), combined with improvements in molecular biology techniques, have improved the study of fungal pathogenicity. Such techniques have provided insights into the infective process by identifying molecular strategies used by the host and pathogen during the course of human mycoses. This chapter will explore the latest discoveries regarding the transcriptome of major human fungal pathogens. Further we will highlight genes essential for host–pathogen interactions, immune response, invasion, infection, antifungal drug response, and resistance. Finally, we will discuss their importance to the discovery of new molecular targets for antifungal drugs. © The Editor(s) (if applicable) and The Author(s), under exclusive license to Springer Nature Switzerland AG 2014, 2022.
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CAUSES OF C AURIS TRANSMISSION In 2009, laboratory personnel isolated C auris from discharge originating in the external ear canal of a hospitalized patient in Japan and named it accordingly.5 Although C auris is closely related to other Candida species, it behaves much differently;rather than originating in the host's flora, it is most often acquired through contact with contaminated surfaces or fomites shed from colonized or infected individuals.6 Patients infected or colonized with C auris shed the pathogen from their skin, which can contaminate the environment-including shared medical equipment.7 Because the pathogen can remain viable on surfaces for prolonged periods (eg, four weeks8), it can be transmitted to patients and lead to systemic infection.9 Traditional biochemical tests for yeast identification may misidentify C auris as another yeast,9,10 and ineffective environmental cleaning can allow it to persist on surfaces, thereby increasing the risk of an outbreak.9 The first reported health care-associated C auris outbreak occurred between April 2015 and July 2016 in a cardiothoracic center in London.11 In June 2016, the CDC issued a clinical alert to health care facilities, noting that one isolate of C auris was detected in 2013 and that the pathogen had been in identified in nine countries on four continents since 2009.12 The number of clinical C auris cases has increased each year since 2016;from 2020 to 2021, that number almost doubled (755 to 1,470).13 In December 2022, there were 1,994 clinical cases and 5,071 screening cases (ie, indicating colonization) of C auris in the United States, with the highest incidence in California, Florida, Illinois, Nevada, New York, and Texas.13 In contrast, from 2013 to 2016, there had only been 63 clinical cases and 14 screening cases reported, occurring only in Illinois, Maryland, New Jersey, and New York. The patient risk factors for acquiring C auris are comparable to those associated with other types of Candida infections, such as Candida albicans, and include * recent surgery,4 * hospitalization in an endemic country outside the United States,4 * extended stays in an intensive care unit,17 * recent care in a postacute setting (eg, long-term care facility),4 * recent therapy with broad-spectrum antimicrobials,4 * the presence of invasive medical devices (eg, central venous catheter, feeding tube, endotracheal tube),4 and * chronic conditions or immunosuppression.18 The risk of healthy individuals (eg, health care workers [HCWs]) contracting C auris is very low. STRATEGIES TO CONSIDER Preventing transmission of epidemiologically important pathogens, such as C auris, in the perioperative practice setting requires strict compliance with infection prevention and control (IPC) measures, including * adhering to hand hygiene requirements, * using transmission-based (ie, contact) precautions, * communicating effectively, and * cleaning and disinfecting the health care environment with approved products.23 In addition, appropriate screening to identify patients colonized or infected with C auris and laboratory surveillance to identify the pathogen also are needed. When caring for patients with possible or confirmed C auris colonization or infection, HCWs should use contact precautions in addition to standard precautions.23 When implementing contact precautions in perioperative areas, personnel should consult with a facility infection preventionist on required personal protective equipment (PPE), patient transport protocol, patient placement in the facility, and enhanced environmental cleaning practices.25 Effective communication may help to ensure that HCWs implement IPC measures correctly and consistently.
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Introduction: Reverse genetics has become an indispensable tool to gain insight into the pathogenesis of viruses and the development of vaccines. The yeast-based synthetic genomics platform has demonstrated the novel capabilities to genetically reconstruct different viruses. Methods: In this study, a transformation-associated recombination (TAR) system in yeast was used to rapidly rescue different strains of feline infectious peritonitis virus, which causes a deadly disease of cats for which there is no effective vaccine. Results and discussion: Using this system, the viruses could be rescued rapidly and stably without multiple cloning steps. Considering its speed and ease of manipulation in virus genome assembly, the reverse genetics system developed in this study will facilitate the research of the feline coronaviruses pathogenetic mechanism and the vaccine development.
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Current agricultural practices, food processing, and extensive availability of unhealthy "fast foods” impose a broad spectrum of degenerative disorders including bacterial, viral, parasitic, fungal, and yeast infections, all of which became a leading cause of death. An infection is defined as "The invasion and growth of germs in the body.” Several of these germs are opportunistic anaerobic organisms, while their propagation and proliferation potentiate the anaerobic bio-environment alarmingly. An ideal therapeutic objective is to restore a healthier cellular "aerobic” metabolic environment. This chapter will extensively focus on three important infectious disorders (i) COVID-19 infection, (ii) Herpesvirus infection, and (iii) Candida albicans yeast infections. Disease etiology, mechanisms, and pathology will be extensively discussed. Prevention and the potential of therapeutic interventions by structurally diverse nutraceuticals, phytopharmaceuticals, probiotics, and micronutrients will be extensively reviewed. © 2023 Elsevier Inc. All rights reserved.
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Background Statins are the cornerstone for the prevention and treatment of atherosclerotic cardiovascular disease (ASCVD). Yet, nearly half of all patients with established ASCVD risk are not on statins and reasons for the pervasive problem of statin nonadherence are not well understood. Artificial intelligence (AI) can be used to analyze social media to provide insights into these barriers and perceptions. Methods We created a pipeline that collects all statin-related discussions from Reddit, a popular social media platform, between 2010 to 2022. Conventional topic modeling techniques were used to cluster these discussions into 100 topics and 6 broad groups. We used a pretrained AI model (RoBERTa) for sentiment analysis to classify each discussion as positive, neutral, or negative. Results We identified 1,189 posts and 9,364 comments related to statins from 5,337 unique users during the study period (Figure). Six key groups of discussion were identified: statin initiation hesitance;clinical trial appraisals;diabetogenic effects and ketogenic diets;common side effects;industry bias around statins;and red yeast rice as alternative. Other notable topics included associations with COVID, coronary calcium, and cognitive decline. Most documents were neutral (66.5%) in tone, but far more were negative (30.8%) than positive (2.6%). Conclusion The use of AI to analyze social media can generate novel insights into perceptions around statins and help guide strategies to improve adherence. [Formula presented]Copyright © 2023 American College of Cardiology Foundation
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Background: The recent emergence of fungal resistance strains has caused concern in medical settings. Medicinal plants continue to be viable sources of bioactive chemicals with therapeutic potential. These compounds can be extracted in different techniques using various solvents that give rise to a wide variety of extracted bioactive compounds that act as anti-fungal. The research aimed to evaluate the effect of fenugreek seed extracts on resistant isolates of Candida spp. isolated from sever COVID-19 patients. Methodology: The study was conducted from August 2021 to November 2022 at Al-Imam Al-Hussein Medical City and Al-Hayat Respiratory Diseases Units. Under a specialist's physician's supervision, severe COVID-19 cases were collected. The collected 455 sputum samples were examined directly and cultured on Sabouraud's Dextrose agar (SDA) media;growth colonies were distinguished and used Grams stain with the API system before the antifungal susceptibility test was performed in accordance with clinical and laboratory standards institute (CLSI 2020) by disc diffusion method to differentiate the resistance microorganism. The extraction process was conducted using the soxhlet technique (100 grams of seed powder and 800 milliliters of solvents (chloroform, methanol, and water) for eight hours. Electrical rotatory evaporators were used to evaporate the extract to get the concentrated crude extracts. FTIR and GC-MS instruments used to detection of bioactive compounds in crude fenugreek seed extracts(aqueous, methanol, and chloroform). Then, different concentrations of each extract (25, 50, 100, and 150 mg/ml) and their effect against the tested resistance study isolated were examined by well diffusion method and Minimum inhibitory concentration was measured. Result(s): A 455 were enrolled in this study. Patients' ages ranged from 20 to 91 years (mean 52.23, SD 15.009). This study indicated that more than half of the samples were males [(262) 57.6%] and [(193) 42.4%] were females. The FTIR and GC-MS showed the methanolic extract potent the most bioactive compounds, followed by the chloroform and water extracts. Evaluation of antimicrobial effects at 50 mg/ml, the methanolic extract showed the greatest effect, with a mean inhibition zone of 9.33 mm and a significant value of 0.01;at 100 mg/ml, the chloroform extract showed the next greatest effect, with a mean inhibition zone of 10.33 mm and a significant value of 0.005. At 150 mg/ml, the aqueous extracts showed the least effect, with a mean inhibition zone of 8.33 mm and a non-significant value of 0.024. Conclusion(s): Candida spp. were most frequent isolated yeast from sputum of patients with severe COVID-19. Methanol extract was the most effective anti-candida, followed by chloroform extract, and the aqueous extract was the least effective. The most effective anti-candida drug is ketoconazole.Copyright © 2022.
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Therapeutic inhibition of critical viral functions is important for curtailing coronavirus disease 2019 (COVID-19). We sought to identify antiviral targets through the genome-wide characterization of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) proteins that are crucial for viral pathogenesis and that cause harmful cytopathogenic effects. All 29 viral proteins were tested in a fission yeast cell-based system using inducible gene expression. Twelve proteins, including eight nonstructural proteins (NSP1, NSP3, NSP4, NSP5, NSP6, NSP13, NSP14, and NSP15) and four accessory proteins (ORF3a, ORF6, ORF7a, and ORF7b), were identified that altered cellular proliferation and integrity and induced cell death. Cell death correlated with the activation of cellular oxidative stress. Of the 12 proteins, ORF3a was chosen for further study in mammalian cells because it plays an important role in viral pathogenesis and its activities are linked to lung tissue damage and a cytokine storm. In human pulmonary and kidney epithelial cells, ORF3a induced cellular oxidative stress associated with apoptosis and necrosis and caused activation of proinflammatory response with production of the cytokines tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), and IFN-ß1, possibly through the activation of nuclear factor kappa B (NF-κB). To further characterize the mechanism, we tested a natural ORF3a Beta variant, Q57H, and a mutant with deletion of the highly conserved residue, ΔG188. Compared with wild-type ORF3a, the ΔG188 variant yielded more robust activation of cellular oxidative stress, cell death, and innate immune response. Since cellular oxidative stress and inflammation contribute to cell death and tissue damage linked to the severity of COVID-19, our findings suggest that ORF3a is a promising, novel therapeutic target against COVID-19. IMPORTANCE The ongoing COVID-19 pandemic caused by SARS-CoV-2 has claimed over 5.5 million lives with more than 300 million people infected worldwide. While vaccines are effective, the emergence of new viral variants could jeopardize vaccine protection. Treatment of COVID-19 by antiviral drugs provides an alternative to battle against the disease. The goal of this study was to identify viral therapeutic targets that can be used in antiviral drug discovery. Utilizing a genome-wide functional analysis in a fission yeast cell-based system, we identified 12 viral candidates, including ORF3a, which cause cellular oxidative stress, inflammation, apoptosis, and necrosis that contribute to cytopathogenicity and COVID-19. Our findings indicate that antiviral agents targeting ORF3a could have a great impact on COVID-19.
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Trichosporon asahii is an opportunistic fungus that forms septate hyphae and pseudohyphae, resembling Candida albicans, and causes fungemia in susceptible individuals. Risk factors for T. asahii infection include immunosuppression, IV catheters, and malignancy. In the present case, a 67-year-old male with a history of renal transplant on immunosuppressive therapy was hospitalized for coronavirus disease 2019 (COVID-19) pneumonia. Despite treatment with steroids and broad initial antibiotic coverage with cefepime, doxycycline, and vancomycin, the patient underwent continual respiratory decline. His sputum culture on hospital day 10 was positive for non-candidal yeast, and despite subsequent appropriate empiric coverage with micafungin and amphotericin B, the patient continued to decline and ultimately died due to the resistance of T. asahii to these antifungals. This case highlights the importance of suspecting T. asahii as an infectious cause in patients whose cultures show non-candidal yeast and initiating appropriate antifungal treatment early in their treatment course.
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Case Report: Over 90% of cases of cryptococcal meningoencephalitis present in immunocompromised patients, with the majority of those being in patients with AIDS. However, this infection can also occur in patients with other immunocompromised states, such as steroid use, malignancy, rheumatologic diseases, and use of immunosuppressive medications. Delay in diagnosis can often lead to rapid neurological deterioration and mortality. Case: A young, otherwise immunocompetent patient, with a history of Chiari I malformation and recent COVID- 19 infection presented with syncope following two weeks of headaches, generalized body aches and weakness after COVID-19 diagnosis. Physical exam demonstrated an isolated CN VI palsy. Head imaging revealed new right caudate infarcts, and a cerebellar tonsillar descent compatible with history of Chiari I malformation. Initial lumbar puncture (LP) was deferred due to congenital brain herniation. Over the next few days, the patient continued to show increasing neurological deficits such as truncal ataxia and increased mood instability. The patient was transferred to the Intensive Care Unit, and LP was obtained under special neuro-critical care direction. Due to increased opening pressures and yeast on gram stain, cryptococcus was suspected and later confirmed. Although anti-fungal therapy was initiated, the patient continued to deteriorate, leading to cardiac arrest, intubation, and placement of lumbar drain. The patient unfortunately did not demonstrate neurologic recovery following arrest and progressed to brain death. Discussion(s): While cryptococcal meningoencephalitis is overwhelmingly a disease of immunocompromised patients, it can occur in immunocompetent hosts, and delay in diagnosis and treatment can lead to adverse and fatal outcomes. This patient had no known underlying conditions besides a recent mild COVID-19 infection and underlying Chiari I malformation, neither of which are known to be associated with cryptococcal meningoencephalitis. These factors may however have played a role in his disease and progression. Copyright © 2023 Southern Society for Clinical Investigation.
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The emergence of severe acute respiratory syndrome coronavirus type II (SARS-CoV-2) variants have led to a decline in the protection of existing vaccines and antibodies, and there is an urgent need for a broad-spectrum vaccination strategy to reduce the pressure on the prevention and control of the pandemic. In this study, the receptor binding domain (RBD) of the SARS-CoV-2 Beta variant was successfully expressed through a glycoengineered yeast platform. To pursue a more broad-spectrum vaccination strategy, RBD-Beta and RBD-wild type were mixed at the ratio of 1:1 with Al(OH)3 and CpG double adjuvants for the immunization of BALB/c mice. This bivalent vaccine stimulated robust conjugated antibody titers and a broader spectrum of neutralizing antibody titers. These results suggested that a bivalent vaccine of RBD-Beta and RBD-wild type could be a possible broad-spectrum vaccination strategy.
ABSTRACT
The emergence of new escape mutants of the SARS-CoV-2 virus has escalated its penetration among the human population and has reinstated its status as a global pandemic. Therefore, developing effective antiviral therapy against emerging SARS-CoV variants and other viruses in a short period of time becomes essential. Blocking SARS-CoV-2 entry into human host cells by disrupting the Spike glycoprotein-angiotensin-converting enzyme 2 (ACE2) interaction has already been exploited for vaccine development and monoclonal antibody therapy. Unlike the previous reports, our study used a nine-amino acid peptide from the receptor-binding motif (RBM) of the Spike (S) protein as an epitope. We report the identification of an efficacious nanobody N1.2 that blocks the entry of pseudovirus-containing SARS-CoV-2 Spike as the surface glycoprotein. Moreover, using mCherry fluorescence based reporter assay we observe a more potent neutralizing effect against both the hCoV19 (Wuhan/WIV04/2019) and the Omicron (BA.1) pseudotyped Spike virus with a bivalent version of the N1.2 nanobody. In summary, our study presents a rapid, and efficient methodology to use peptide sequences from a protein-receptor interaction interface as epitopes for screening nanobodies against potential pathogenic targets. We propose that this approach can also be widely extended to target other viruses and pathogens in the future.