Exposure to SARS-CoV-2 Aerosols in U.S. Residences: Sample analysis Using Two Detection Criteria

Uniform practices and quality control methods are needed to detect and quantify airborne viruses across sampling and analysis platforms. We compared detection of airborne SARSCoV-2 RNA in residences of individuals with COVID-19 using two commonly used criteria: environmental (at least one SARS-CoV-2-specific gene and internal control amplified by PCR with Ct ≤ 40) and clinical (at least two SARS-CoV-2-specific genes and internal control amplified with Ct ≤ 37). 24-hr total aerosol samples were collected in a self-isolation room and an additional room without manipulating subjects' behavior/activities. Under the environmental criterion, 7/16 samples in primary rooms and 7/15 samples in secondary rooms were positive. Comparable but lower positive sample proportions were observed using the more rigorous clinical criterion: 6/16 primary rooms and 5/15 secondary rooms. A consensus SARS-CoV-2 environmental sampling and analysis framework is needed for comparisons between studies. © 2022 17th International Conference on Indoor Air Quality and Climate, INDOOR AIR 2022. All rights reserved.

Sirna-A Potential Therapy for Covid-19 Infection

Background: The deadly virus COVID-19 has affected more than 1 crore people and claimed more than 5 lakh lives worldwide according to the World health organization. Though there are numerous treatment modalities available including anti-bacterials, antivirals, vaccines etc., none of them can be considered as effective cure for SARS CoV-2 virus as they are mostly non-specific in action. Aim(s): siRNA therapy can be considered as a significant treatment modality due to its specificity in action. The aim of this review is to explore siRNA as a potential treatment strategy for the treatment of COVID-19. Material(s) and Method(s): In this review we shall explore the targets of siRNA therapy which includes viral RNA-dependent RNA polymerase, helicase, protease and nucleoprotein N. siRNA related patents provide solutions for novel RNAi techniques, high expense of chemically synthetic siRNA, techniques for restraining SARS-CoV by disturbing RNA etc., siRNA-based drug delivery systems and limitations of nanocarrier delivery system were reviewed. siRNA is a gene silencer that targets highly conserved sequences which codes for protease 3CL (nsp5) and viral helicase (from 16-18 kbp). Conclusion(s): Thus, siRNA-based therapy is considered highly efficacious as it can hit the highly conserved regions of SARS-CoV-2 RNA.Copyright © 2023, Advanced Scientific Research. All rights reserved.

Jahrestagung der Gesellschaft fur Padiatrische Pneumologie.

The proceedings contain 54 papers. The topics discussed include: cytokines in severe childhood asthma;transcriptional gene regulation of interleukin-6 in epithelial cells in viral-induced asthma exacerbation;assessment of the long-term safety and efficacy of dupilumab in children with asthma: LIBERTY ASTHMA EXCURSION;impulse oscillometry bronchodilator response in preschool children;pulmonary function in non-hospitalized adults and children after mild Covid-19;exhaled aerosols in PCR-confirmed SARS-CoV-2-infected children;early respiratory infectious diseases have an influence on the gut microbiome;comparison of three eradication treatment protocols for pseudomonas aeruginosa in children and adolescents with cystic fibrosis;neutrophilic airway inflammation in children with repaired esophageal atresia-tracheoesophageal fistula (EA/TEF);and multiplex immunofluorescence and multispectral imaging as a tool to evaluate host directed therapy.

Pyrrole-Imidazole Polyamides - A Frontrunner in Nucleic Acid-Based Small Molecule Drugs

The remarkable success of messenger RNA vaccines against the ongoing coronavirus-2019 (COVID-19) pandemic renews attention toward nucleic acid therapeutics. While nucleic acid therapy using unmodified DNA or RNA is the primary focus in disease treatment, there is growing need to develop nucleic acid-based small molecules owing to their potential clinical benefits as drugs in terms of cost and scalability. While small molecules targeting protein-protein interactions are known to alter the transcriptional status of a cell, they can result in a transient effect and variation of bio-efficacy among patients. Small molecules targeting DNA and/or RNA are in demand in the precision medicine approach as they have consistent bioactivity among patients. This review details the progress of sequence-specific DNA-binding pyrrole-imidazole polyamides (PIPs) in modulating the transcriptional status of target gene(s) without altering the underlying DNA sequence. Here, the different versions of PIPs are listed, and also, how conjugating them with DNA alkylating agents, epigenetic modulators, and other drugs can improve their clinical utility as targeted transcription therapeutics. Owing to their specificity, functional diversity, and limited toxicity, PIP technology holds enormous promise as frontrunner in small-molecule-based nucleic acid drugs to precisely regulate therapeutically important genes on demand and treat intractable diseases.Copyright © 2023 Wiley-VCH GmbH.

RNA INTERFERENCE IS THE BASIS OF HUMAN ANTIVIRAL DEFENSE.

Comparative analysis of antiviral protective mechanisms in protozoa and RNA interference of multicellular organisms has revealed their similarity, also providing a clue to understanding the adaptive immunity. In this article, we present the latest evidence on the importance of RNA-guided gene regulation in human antiviral defense. The role of neutralizing antibodies and interferon system in viral invasion is considered. The new concept has been introduced, i.e., antiviral protection of any living organism is based on the intracellular RNA-guided mechanisms. Simple and effective defense against viruses is that spacer segment of the viral DNA is inserted into the cellular chromosomes. Upon re-infection, the RNA transcript of the spacer directs nuclease enzymes against the foreign genome. This is a really adaptive immune defense that any cell potentially possesses. In humans, the interferon system provides an additional tool for early suppression of viral infections which shifts the cells to the alert regimen, thus preventing further spread of infection. The main task of the human central immune system is to maintain integrity and combat foreign organisms. Accordingly, a suitable index of acquired antiviral immunity should be a presence of specific spacer markers in DNA samples from reconvalescent persons, rather than detection of neutralizing antibodies, B and T memory cells. This article is addressed primarily to general medical community, and its practical conclusions are as follows: 1. Presence or absence of specific antibodies to SARS-CoV-2 is not a prognostic sign of the disease. Detection of specific antibodies in blood simply reflects the fact that the person has contacted with the viral agent. Absence of antibodies does not mean a lack of such contact, and the persons with high titers of specific antibodies are not protected from re-infection with SARS-CoV-2. 2. PCR testing: The PCR results may remain "false positive" in those subjects who have had COVID-19, if the genetic material is taken from the site of initial virus contraction (mainly, nasopharynx). In our opinion, negative PCR tests for COVID-19 in blood plasma and urine will be a more correct index for the absence of the disease, even with positive PCR tests from the nasopharyngeal samples. 3. It is necessary to draw attention of general practitioners to potential usage of retinol in prevention and treatment of COVID-19, given the importance of RLR receptors in recognition of viral RNAs and positive experience of vitamin A administration in measles, another dangerous viral disease. Copyright © 2022, SPb RAACI.

Extending and adapting the functions of genetic laboratories during the COVID-19 pandemic–challenges and successes

Clinical molecular genetics laboratories have expanded rapidly in the last 15 years, incorporating new technologists at an astounding rate that has brought rare disease testing out of research labs and into standard of care medical practice. These laboratories have had to adapt a succession of new technologies and methods of data analysis while building in-house expertise. When the SARS-CoV-2 virus, the cause of COVID-19, emerged in early 2020 and quickly spread across the globe, many areas of the United States (U.S.) when into lockdown. Noncritical healthcare appointments were postponed resulting in a dramatic drop in the number of samples being referred for genetic testing for rare diseases. As large genetics laboratory experienced the resulting drop in volume, the demand for SARS-CoV-2 testing soared. Equipped with expertise in high throughput testing, as well as clinical technologists trained in high-complexity testing, large genetics laboratories stepped in the fill the gap, a measure that kept laboratories running and staff employed. Our expertise in highthroughput high-complexity led from requests to perform testing in in our genomics laboratory to building new laboratories in both the U.S. and the United Kingdom (U.K.). These efforts resulted in building three laboratories from an empty space to a functioning, staffed clinical laboratory in approximately eight weeks. These laboratories employ over 1200 individuals (∼550 U.S. and ∼700 U.K.) with plans to expand to over 2000. To date, these laboratories have performed ∼2.5 million SARS-CoV-2 assays. Challenges included navigating state, federal, and country regulations and rapidly training a large clinical staff while ensuring optimal assay performance. Clinical testing in the U.S. is governed by the Clinical Laboratory Improvement Amendments (CLIA), which provide very specific requirements for personnel, training, proficiency testing and the quality management system. However, high complexity molecular testing for a viral target could fall into the CLIA category of general chemistry (as does molecular genetic testing) or microbiology, subcategory virology. The category chosen has dramatic effects on the specific experience required for technologists, supervisors and the laboratory director. Outside the U.S., laboratory requirements are dictated by accepted best practices and accrediting agencies, rather than specific laws, sometimes making it difficult to know what requirements need to be met. Various assays with slightly different designs are available, and the assay used must be best suited to the testing workflow. In the U.S., samples collection is supervised by a healthcare provider. A higher sensitivity assay that does not include an internal human control genewas chosen. In the U. K., home collection is allowed, therefore, an assay that includes a human RNAseP gene control but with lower sensitivity for SARS-CoV- 2 was chosen. Given the current global awareness of respiratory virus activity and spread, there is a growing demand for newand expanded testing. Combining SARS-CoV-2 testing with influenza, RSV and potentially other viruses is clinically desirable. Pooling of samples will allow for even greater throughput while reducing the demand for increasingly scarce consumables. Finally, our experience with highthroughput sequencing is allowing us to pivot quickly to viral genome sequencing, which is proving critical to understanding and combating this pandemic. Rare metabolic diseases, intellectual disabilities and hereditary cancer syndromes will always still need attention and continuous innovation. We will need to learn to balance these activities and continue to support testing needs for these in addition to emerging diseases.