COVID-19 complicating pregnancy: A retrospective study of nine intensive care unit patients.

Background: With the emergence of the coronavirus 2019 (COVID-19) pandemic, it was essential to determine the impact of this disease on pregnant women and neonatal outcomes. In this study, we present a series of nine cases of pregnant women with COVID-19 disease requiring intensive care unit (ICU) admission. Methods: We retrospectively collected clinical data of pregnant women with COVID-19 disease admitted to ICU between September 2020 and September 2021. Results: Most common presenting symptom was cough. Two patients had no respiratory symptoms at presentation. Five of the nine patients required invasive mechanical ventilation. Seven patients required caesarean section, four of whom delivered preterm. There were no maternal or neonatal deaths. Conclusions: Although maternal and neonatal outcomes reported in our study are encouraging, it is imperative to emphasize the importance of an individualized, multidisciplinary approach, and good healthcare infrastructure for optimal management of this group of patients.

Insertion of foreign genes into the simian varicella virus genome by Tn7-mediated site-specific transposition.

A Tn7-transposition approach was utilized for site-specific insertion of foreign genes into the genome of simian varicella virus (SVV), the causative agent of simian varicella in nonhuman primates. The severe acute respiratory syndrome coronavirus (SARS-CoV-2) nucleocapsid (N) gene and receptor binding domain (RBD) of the spike gene were inserted into the ORF 14 region of the SVV genome cloned into a bacterial artificial chromosome and then transfected into Vero cells to generate the infectious recombinant SVV (rSVV). The rSVV replicated efficiently in infected Vero cells and expressed the N and RBD antigens as indicated by immunoblot and immunofluorescence assays. Tn7-mediated transposition provides a rapid and efficient method for constructing rSVVs which may be evaluated as live-attenuated vaccines.

LAMPrimers iQ: New primer design software for loop-mediated isothermal amplification (LAMP).

Nucleic acids amplification is a widely used technique utilized for different manipulations with DNA and RNA. Although, polymerase chain reaction (PCR) remains the most popular amplification method, isothermal approaches are gained more attention last decades. Among these, loop-mediated isothermal amplification (LAMP) became an excellent alternative to PCR. LAMP requires an increased number of primers and, therefore, is considered a highly specific amplification reaction compared to PCR. LAMP primers design is still a non-trivial task, and all niceties should be taken into account during their selection. Here, we report on a new program called LAMPrimers iQ destined for high-quality LAMP primers design. LAMPrimers iQ is based on an original algorithm considering rigorous criteria for primers selection. Unlike alternative programs, LAMPrimers iQ can process long DNA or RNA sequences, and completely avoid primers that can form homo- and heterodimers. The quality of the primers designed was checked using SARS-CoV-2 coronavirus RNA as a model target. It was shown that primers selected with LAMPrimers iQ provide higher specificity and reliable detection of viral RNA compared to those obtained by alternative programs. The program is available at https://github.com/Restily/LAMPrimers-iQ.

Development of potent and effective SARS-CoV-2 main protease inhibitors based on maleimide analogs for the potential treatment of COVID-19.

In the present work, we report a new series of potent SARS-CoV-2 Main Protease (Mpro) inhibitors based on maleimide derivatives. The inhibitory activities were tested in an enzymatic assay using recombinant Mpro (3CL Protease from coronavirus SARS-CoV-2). Within the set of new Mpro inhibitors, 6e demonstrated the highest activity in the enzymatic assay with an IC50 value of 8.52 ± 0.44 µM. The IC50 value for Nirmatrelvir (PF-07321332, used as a reference) was 0.84 ± 0.37 µM. The cytotoxic properties were determined in the MTT assay using MRC-5 and HEK-293 cell lines. In the course of the investigation, we found that the newly obtained maleimide derivatives are not substantially cytotoxic (IC50 values for most compounds were above 200 µM).

Membrane activity of viral proteins.

COVID-19 infection, caused by the SARS-CoV-2 coronavirus, has led to the largest pandemic since the Spanish flu in 1918. In view of this, the development of vaccines and antiviral drugs that can stop the spread of this infection has become an acute issue. Currently, in the search for antiviral drugs against COVID-19, much attention is paid to the study of the structure of the receptor-binding domain of the surface protein S. However, the emergence of new SARS-CoV-2 coronavirus strains indicates its high variability, which reduces the effectiveness of vaccines and antiviral drugs. At the same time, the envelope protein E of this virus is membrane active and shows a rather high conservatism. Despite the critical importance of this protein in the coronavirus life cycle, the physicochemical mechanisms of its interaction with cell membranes still remain unclear. So, we investigated the membrane activity of protein E of the SARS-CoV-2 coronavirus on models of giant unilamellar vesicles and lipid nanotubes. As a result, it was found that the protein forms pores in the lipid bilayer, i.e., performs the main function of viroporin. In addition, protein E is able to deform lipid membranes and form double-membrane vesicles depending on the concentration.

The influence of SARS-CoV-2 infection on the quality of life of women who reach menopause.

Introduction: At the time of the COVID-19 pandemic, providing adequate medical care in all its aspects, including the care of women with menopause and keeping social distance, was a challenge. Menopause results in a lower level of oestrogens and progesterone, which is the cause of lower immunological response and may result in more people being ill with COVID-19. The aim of the research was to evaluate the correlation between being sick with COVID-19 and the quality of life of women with menopause. Material and methods: The research was done in a group of 249 women with menopause. The criteria deciding about inclusion into the group were as follows: female gender, age 40-65 years, time after infection with SARS-CoV-2 virus 14-30 days, no hospitalization, and diagnosis of SARS-CoV-2 virus infection by means of anti-gene test. A propriety survey was used as well as medical documents analysis and a questionnaire with standardized WHOQOL-BREF. SPSS Statistics 27.0 program was used for statistical analysis. In all calculations p < 0.05 was accepted as the level of significance. Results: While evaluating the quality of life in the case of women after suffering from COVID-19 caused by the SARS-CoV-2 virus, no statistically significant difference was observed. The correlation between the level of satisfaction with one's health and suffering from SARS-CoV-2 was within the range of α = 0.1, with a significance level p = 0.061. Conclusions: No statistically significant correlation was noted between the quality of life of women with menopause after SARS-CoV-2 and women who did not suffer from it.

Higher Rates of Complicated Appendicitis During the COVID-19 Pandemic: A Year-to-Year Analysis.

INTRODUCTION: The COVID-19 pandemic impacted presentation, management strategies, and patient outcomes of numerous medical conditions. The aim of this study is to perform a year-to-year comparison of clinical outcomes of patients with acute appendicitis (AA) before and during the pandemic. METHODS: Patients treated for AA during the initial 12-mo period of the pandemic at our institute were compared to those treated for AA during the 12-mo period before. Clinical and laboratory parameters, treatment strategies, intraoperative findings, pathology reports, and postoperative outcomes were compared. RESULTS: During the study period, 541 patients presented with AA. The median (interquartile range) age was 28 (21-40) y and 292 (54%) were males. 262 (48%) patients presented during the pre-COVID-19 period, while 279 patients (52%) presented during the COVID 19 pandemic. The groups were comparable for baseline clinical data and imaging results upon index admission. There was no significant difference in rate of nonoperative treatment between the Pre-COVID-19 and During-COVID-19 eras (51% versus 53%, P = 0.6) as well as the success rate of such treatment (95.4% versus 96.4%, P = 0.3). Significantly more patients presented with a periappendicular abscess during COVID-19 (4.6% versus 1.1%, P = 0.01) and median (interquartile range) operative time was significantly longer (78 (61-90) versus 32.5 (27-45) min, P < 0.001). Pathology reports revealed a higher rate of perforated appendicitis during COVID-19 (27.4% versus 10.2%, P < 0.001). CONCLUSIONS: Patients with AA present with higher rates of perforated and complicated appendicitis during the COVID-19 pandemic. The success rates of nonoperative management in selected patients with noncomplicated AA did not change during the pandemic and is a safe, feasible, option.

Detection of Specific RNA Targets by Multimerization.

Detection of specific RNA targets via amplification-mediated techniques is widely used in fundamental studies and medicine due to essential role of RNA in transfer of genetic information and development of diseases. Here, we report on an approach for detection of RNA targets based on the particular type of isothermal amplification, namely, reaction of nucleic acid multimerization. The proposed technique requires only a single DNA polymerase possessing reverse transcriptase, DNA-dependent DNA polymerase, and strand-displacement activities. Reaction conditions that lead to efficient detection of the target RNAs through multimerization mechanism were determined. The approach was verified by using genetic material of the SARS-CoV-2 coronavirus as a model viral RNA. Reaction of multimerization allowed to differentiate the SARS-CoV-2 RNA-positive samples from the SARS-CoV-2 negative samples with high reliability. The proposed technique allows detection of RNA even in the samples, which were subjected to multiple freezing-thawing cycles.

Ferrans and Powers Quality of Life Index Applied to Nursing Students during the COVID-19 Pandemic / Índice de Qualidade de Vida de Ferrans e Powers Aplicado a Estudantes de Enfermagem Durante a Pandemia de COVID-19

COVID-19 pandemic can have a significant impact on an individual's physical and emotional well-being. However, it is important to note that not all experiences of the pandemic are negative. This study aims to evaluate the perception of quality of life of students enrolled in a nursing course during the COVID-19 pandemic by administering a global questionnaire. A quantitative, exploratory, cross-sectional, and descriptive study to investigate perception of quality of life of students during the COVID-19 pandemic. Participants comprised 80 students graduating with a nursing qualification between 2020 and 2021. The Ferrans and Powers Quality of Life Index was used in the study. The mean scores are reported as follows: health/functioning domain (22.06), socioeconomic domain (21.40), psychological/spiritual domain (23.34), and family domain (23.06), with an average general quality of life index of 22.46. After evaluating the domains, it was found that there was a higher perception of the psychological/spiritual domain and lower perception of the socioeconomic domain. The results supported the internal consistency reliability of the entire Quality of Life (QLI; alpha=0.95) and the four subscales (alpha=0.88, 0.71, 0.92, and 0.80). The study suggests that the COVID-19 pandemic did not have an impact on the perceived quality of life among students.(AU)

Pandemia de COVID-19 pode ter um impacto significativo no bem-estar físico e emocional de um sujeito. No entanto, é importante notar que nem todas as experiências da pandemia são negativas. Este estudo teve como objetivo avaliar a percepção da qualidade de vida de estudantes matriculados em um curso de enfermagem durante a pandemia de COVID-19 por meio da aplicação de um questionário global. Estudo quantitativo, exploratório, transversal e descritivo para investigar a percepção da qualidade de vida de escolares durante a pandemia de COVID-19. Participaram 80 estudantes concluintes do curso de enfermagem entre 2020 e 2021. O Índice de Qualidade de Vida de Ferrans e Powers foi utilizado no estudo. Os escores médios foram relatados da seguinte forma: domínio saúde/funcionamento (22,06), domínio socioeconômico (21,40), domínio psicológico/espiritual (23,34) e domínio família (23,06), com média geral do índice de qualidade de vida de 22,46. Após a avaliação dos domínios, constatou-se maior percepção do domínio psicológico/espiritual e menor percepção do domínio socioeconômico. Os resultados apoiaram a confiabilidade da consistência interna de toda a Qualidade de Vida (QLI; alfa=0,95) e as quatro subescalas (alfa=0,88, 0,71, 0,92 e 0,80). O estudo sugere que a pandemia de COVID-19 não teve impacto na qualidade de vida percebida entre os estudantes.(AU)

Pyridine Derivatives as Potential Inhibitors for Coronavirus SARS-CoV-2: A Molecular Docking Study.

Coronavirus SARS-CoV-2, a causative agent for the global epidemic disease COVID-19, which has a highest modality rate. Several initiatives have been undertaken to repurpose current antiviral medications and tested the classic pyridine derivatives (PyDev), which have showed substantial therapeutic potential against a variety of illnesses and also have several biological functions such as, antibacterial, antiviral, and anti-inflammatory. However, limited reports are available for the treatment of Coronavirus SARS-CoV-2 using PyDev. Hence, the possibilities of the best-described PyDev molecules of powerful Coronavirus SARS-CoV-2 inhibitors have been attempted in this investigation. This study primarily focused on blocking four key targets of Coronavirus SARS-CoV-2 proteins. Terpyridine has shown the greatest inhibitory potential (with a binding energy of -8.8 kcal/mol) against all four coronavirus targets. This study results would pave the potential lead medication for Coronavirus SARS-CoV-2 therapeutic strategies.

Possibilities of simulation of coronavirus SARS-CoV-2 by using light scattering approach.

Issues related to human coronavirus (SARS CoV-2) are a burning topic of research in present times. Due to its easily contagious nature, real experimentation under laboratory conditions requires a high level of biosafety. A powerful algorithm serves as a potential tool for the analysis of these particles. We attempted to simulate the light scattering from coronavirus (SARS CoV-2) model. Different images were modelled using a modified version of a Monte Carlo code. The results indicate that spikes on the viruses exhibit a significant scattering profile and that the presence of spikes during modelling contributes to the distinctiveness of the scattering profiles.

Wave of COVID-19 outbreaks in Gold Coast residential aged care facilities after easing travel restrictions into Queensland.

Objective: This paper describes outbreaks of coronavirus disease 2019 (COVID-19) in Gold Coast residential aged care facilities (RACFs), in the two months following the easing of travel restrictions at Queensland's domestic border on 13 December 2021. Methods: This audit reviewed all RACF COVID-19 outbreaks notified to the Gold Coast Public Health Unit between 13 December 2021 and 12 February 2022. An outbreak was defined by the Communicable Diseases Network Australia guidelines current at the time. Results: There were 60 COVID-19 outbreaks across 57 RACFs during this period. In 44 outbreaks (73.3%), a staff member was identified as the primary or co-primary case. Transmission amongst residents occurred in 48 outbreaks (80.0%). The attack rates in staff and residents were 17.0% (n = 1,060) and 11.7% (n = 645) respectively. A higher number of males were hospitalised (n = 39: 57.4%) or died (n = 28: 66.7%) than were females (n = 29: 42.6%; n = 14: 33.3% respectively). Most resident cases (n = 565: 87.6%) had received two or more doses of a COVID-19 vaccine. In resident cases who were under-vaccinated (n = 76), twenty (26.3%) required hospitalisation and nine (11.8%) died. In resident cases who received two doses of vaccine (n = 484), forty-three (8.9%) were hospitalised and 27 (5.8%) died. In resident cases who had received three doses (n = 80), four (5.0%) were hospitalised and five (6.3%) died. Conclusions: COVID-19 caused significant morbidity and mortality in Gold Coast RACFs following the easing of border restrictions. Higher rates of hospitalisation and death occurred in males than in females, and in under-vaccinated resident cases than in those vaccinated with at least two doses of a COVID-19 vaccine. Implications for public health: These data support the need for up-to-date COVID-19 vaccination of residents in RACFs, continued surveillance and timely and appropriate implementation of public health guidelines to manage COVID-19 outbreaks in RACFs.

Monoclonal antibody designed for SARS-nCoV-2 spike protein of receptor binding domain on antigenic targeted epitopes for inhibition to prevent viral entry.

SARS, or severe acute respiratory syndrome, is caused by a novel coronavirus (COVID-19). This situation has compelled many pharmaceutical R&D companies and public health research sectors to focus their efforts on developing effective therapeutics. SARS-nCoV-2 was chosen as a protein spike to targeted monoclonal antibodies and therapeutics for prevention and treatment. Deep mutational scanning created a monoclonal antibody to characterize the effects of mutations in a variable antibody fragment based on its expression levels, specificity, stability, and affinity for specific antigenic conserved epitopes to the Spike-S-Receptor Binding Domain (RBD). Improved contacts between Fv light and heavy chains and the targeted antigens of RBD could result in a highly potent neutralizing antibody (NAbs) response as well as cross-protection against other SARS-nCoV-2 strains. It undergoes multipoint core mutations that combine enhancing mutations, resulting in increased binding affinity and significantly increased stability between RBD and antibody. In addition, we improved. Structures of variable fragment (Fv) complexed with the RBD of Spike protein were subjected to our established in-silico antibody-engineering platform to obtain enhanced binding affinity to SARS-nCoV-2 and develop ability profiling. We found that the size and three-dimensional shape of epitopes significantly impacted the activity of antibodies produced against the RBD of Spike protein. Overall, because of the conformational changes between RBD and hACE2, it prevents viral entry. As a result of this in-silico study, the designed antibody can be used as a promising therapeutic strategy to treat COVID-19.

Humoral efficacy of the third SARS-CoV-2 vaccine dose in Multiple Sclerosis subjects undergoing different disease-modifying therapies.

BACKGROUND: It remains unclear how vaccine doses and combinations of vaccination and infection affect the magnitude and quality of immune responses, particularly against novel SARS-CoV-2 variants in subjects with immune-related disorders, such as people with multiple sclerosis (pwMS). Several studies have evaluated the duration of anti-SARS-CoV-2 immune protection in healthy individuals; however clinical data suggest an attenuated short-term humoral response to SARS-CoV-2 vaccines in pwMS receiving disease-modifying therapies (DMTs). METHODS: In this prospective study, we evaluated the humoral response to the third (3rd) BNT162b2 vaccine (booster) dose in a monocentric cohort of pwMS undergoing eight different DMTs, all without previous SARS-CoV-2 infection. Quantitative determination of SARS-CoV-2 IgG Spike titre was carried out by anti-SARS-CoV-2 S assay in 65 pwMS and 9 healthy controls, all without previous SARS-CoV-2 infection. Moreover, these measurements were also compared to their relative levels at 21 days (T1) and ∼6 months (T2) after the second (2nd) vaccination. RESULTS: We observed that the humoral response to the booster dose in Interferon ß-1a-, Dimethyl fumarate- and Teriflunomide-treated pwMS is comparable to healthy controls, while increased in Cladribine-treated pwMS. Additionally, the 3rd dose elicits a seroconversion in the 100% of pwMS under Fingolimod and in the 65% of those under Ocrelizumab. Moreover, multivariate regression analysis showed that treatment with Interferon ß-1a, Dimethyl fumarate and Cladribine positively associates with an increased humoral response. CONCLUSIONS: Taken together this evidence strongly indicates the importance of the booster dose to enhance SARS-CoV-2-specific immunity especially in immunocompromised subjects, such as pwMS under DMTs.

Salud mental en tiempos de pandemia / Mental health in pandemic times

La enfermedad por coronavirus de 2019 (COVID-19) es una emergencia de salud pública catalogada por la Organización Mundial de la Salud como pandemia, la cual ha generado múltiples síntomas deletéreos en la salud mental a escala global, derivados, en parte, de la incertidumbre sobre la calidad y precisión de los informes ofrecidos al respecto y, en gran medida, por la imposibilidad de saber cuándo el mundo volverá a la normalidad. Por tales razones, con este manuscrito se pretende revelar la importancia de la salud mental de la población durante la pandemia, para lo cual se brindan estrategias de prevención e intervención que contribuyan a superar los desafíos propios a los cuales se han enfrentado los individuos y que inevitablemente, en la mayoría de los casos, han sido abordados de forma inadecuada.

Coronavirus disease 2019 (COVID-19) is an emergency of public health classified by the World Health Organization as pandemic, which has generated multiple deleterious symptoms in mental health at global scale, derived, partly, of the uncertainty about the quality and precision of the reports offered in this respect and, in great extent, for the impossibility of knowing when the world will return to normality. For such reasons, with this manuscript it is sought to reveal the importance of the population mental health during the pandemic, for which strategies of prevention and intervention are offered that contribute the challenges faced by individuals and that inevitably, in most of the cases, have been approached in an inadequate way.

Early laboratory hematological parameters associated with COVID-19 mortality in an Amazonian population of Peru

Introduction: COVID-19 has a significant impact on the hematopoietic system and hemostasis. Leukocytosis, lymphopenia, and thrombocytopenia are associated with increased severity and even death in COVID-19 cases. Objective: The aim is to examine the laboratory results of COVID-19 patients from a hospital in the Peruvian Amazon and their clinical prognosis. Material and Methods: An analytical cross-sectional study was carried out whose purpose was to identify the laboratory tests of patients with COVID-19 and mortality in a hospital in Ucayali, Peru during the period from March 13 to May 9, 2020, selecting a total of 127 with Covid-19. Mean and the standard deviation was described for age, leukocytes, neutrophils, platelets, RDW-SD; median and interquartile range for the variables lymphocyte, RN / L, fibrinogen, CRP, D-dimer, DHL, hematocrit, monocytes, eosinophils. Results: No differences were observed in this population regarding death and sex (OR: 1.31; 95% CI 0.92 to 1.87), however, it was observed that, for each one-year increase, the probability of death increased by 4% (PR: 1.04, 95% CI 1.03 to 1.05). The IRR (Incidence Risk Ratio) analysis for the numerical variables showed results strongly associated with hematological values such as Leukocytes (scaled by 2500 units) (IRR: 1.08, 95% CI 1.03 to 1.13), neutrophils (scaled by 2500 units) (IRR: 1.08; 95% CI 1.03 to 1.13), on the contrary, it is observed that the increase of 1000 units in lymphocytes, the probability of dying decreased by 48% (IRR: 0.52; 95% CI 0.38 to 071). Conclusions: Parameters such as leukocytes,neutrophils and D-dimer were statistically much higher in patients who died.

Introducción: COVID-19 tiene un impacto significativo en el sistema hematopoyético y la hemostasia. La leucocitosis, la linfopenia y la trombocitopenia se asocian con una mayor gravedad e incluso la muerte en los casos de COVID-19. Objetivo: examinar los resultados de laboratorio de pacientes con COVID-19 de un hospital de la Amazonía peruana y su pronóstico clínico. Material y métodos: Se realizó un estudio transversal analítico cuyo propósito fue identificar las pruebas de laboratorio de pacientes con COVID-19 y mortalidad en un hospital de Ucayali, Perú durante el periodo del 13 de marzo al 9 de mayo del 2020, seleccionando un total de 127 con COVID-19. Se describió la media y la desviación estándar para edad, leucocitos, neutrófilos, plaquetas, RDW-SD; mediana y rango intercuartílico para las variables linfocito, RN/L, fibrinógeno, PCR, dímero D, DHL, hematocrito, monocitos, eosinófilos. Resultados: No se observaron diferencias en esta población en cuanto a muerte y sexo (OR: 1,31; IC 95% 0,92 a 1,87), sin embargo, se observó que, por cada aumento de un año, la probabilidad de muerte aumentaba un 4% (RP: 1,04). , IC del 95%: 1,03 a 1,05). El análisis de RIR (Razón de incidencia de riesgos) para las variables numéricas mostró resultados fuertemente asociados con valores hematológicos como Leucocitos (escala de 2500 unidades) (RRI: 1.08, 95% CI 1.03 a 1.13), neutrófilos (escala de 2500 unidades) (RRI: 1.08; IC 95% 1.03 a 1.13), por el contrario, se observa que al aumento de 1000 unidades en linfocitos, la probabilidad de morir disminuyó en un 48% (TIR: 0.52; IC 95% 0.38 a 071). Conclusiones: Parámetros tales como los leucocitos, los neutrófilos y el dímero D fueron estadísticamente mucho más altos en los pacientes que fallecieron.

El enigma del coronavirus - Reinfecciones por COVID-19, COVID prolongado - Vacunas bivalentes La gestante - Derechos humanos y Ética

Al finalizar el segundo año de la pandemia COVID-19, da la impresión que el coronavirus enigmático nos estuviera otorgando cierto respiro. Sin embargo, las cifras de infectados han iniciado un ascenso en el hemisferio norte, donde se aproxima un invierno fuerte, ya con nevadas. Pero también se observa este nuevo incremento en el hemisferio sur, como en el Perú, cuando estamos en primavera. El SARS-CoV-2 continúa diferenciándose en multitud de variantes y subvariantes, algunas de la cuales tienen mayor facilidad de evadir la inmunidad humana y aquella alcanzada con las vacunas de ARNm, y también podrían no responder a los tratamientos con anticuerpos monoclonales. La información clínica actual está orientada a la expectativa sobre si las vacunas existentes pudieran al menos minorizar las hospitalizaciones, el ingreso a cuidados intensivos y las muertes. Se sigue aprendiendo sobre los efectos clínicos de la infección por el SARS-CoV-2, la ocurrencia de reinfecciones y los daños físicos y mentales a largo plazo, buscándose la prevención de la infección, las reinfecciones y el COVID prolongado.

At the end of the second year of the COVID-19 pandemic, it seems that the enigmatic coronavirus is giving us some respite. However, the number of people infected has started to rise in the northern hemisphere, where a strong winter with snowfall is approaching. But this new increase is also observed in the southern hemisphere, as in Peru, when we are in spring. SARS-CoV-2 continues to differentiate into a multitude of variants and subvariants, some of which are more easily able to evade human immunity and that achieved with mRNA vaccines, and also may not respond to monoclonal antibody treatments. Current clinical information is oriented to the expectation that existing vaccines could at least reduce hospitalizations, intensive care admission and deaths. Learning about the clinical effects of SARS-CoV-2 infection, the occurrence of reinfection, and long-term physical and mental harm continues, with prevention of infection, reinfection, and prolonged COVID being sought.

A Novel Rolling Circle Amplification-Based Detection of SARS-CoV-2 with Multi-Region Padlock Hybridization.

SARS-CoV-2 has remained a global health burden, primarily due to the continuous evolution of different mutant strains. These mutations present challenges to the detection of the virus, as the target genes of qPCR, the standard diagnostic method, may possess sequence alterations. In this study, we develop an isothermal one-step detection method using rolling circle amplification (RCA) for SARS-CoV-2. This novel strategy utilizes a multi-padlock (MP-RCA) approach to detect viral-RNA via a simplified procedure with the reliable detection of mutated strains over other procedures. We designed 40 padlock-based probes to target different sequences across the SARS-CoV-2 genome. We established an optimal one-step isothermal reaction protocol utilizing a fluorescent output detected via a plate reader to test a variety of padlock combinations. This method was tested on RNA samples collected from nasal swabs and validated via PCR. S-gene target failure (SGTF)-mutated strains of SARS-CoV-2 were included. We demonstrated that the sensitivity of our assay was linearly proportional to the number of padlock probes used. With the 40-padlock combination the MP-RCA assay was able to correctly detect 45 out 55 positive samples (81.8% efficiency). This included 10 samples with SGTF mutations which we were able to detect as positive with 100% efficiency. We found that the MP-RCA approach improves the sensitivity of the MP-RCA assay, and critically, allows for the detection of SARS-CoV-2 variants with SGTF. Our method offers the simplicity of the reaction and requires basic equipment compared to standard qPCR. This method provides an alternative approach to overcome the challenges of detecting SARS-CoV-2 and other rapidly mutating viruses.

Phytochemical Discrimination, Biological Activity and Molecular Docking of Water-Soluble Inhibitors from Saussurea costus Herb against Main Protease of SARS-CoV-2.

Siddha medicine is one of the oldest medical systems in the world and is believed to have originated more than 10,000 years ago and is prevalent across ancient Tamil land. It is undeniable that inhibitor preferences rise with increasing solubility in water due to the considerations pertaining to the bioavailability and the ease of which unabsorbed residues can be disposed of. In this study, we showed the phytochemical discrimination of Saussurea costus extracted with water at room temperature as a green extraction procedure. A total of 48 compounds were identified using gas chromatography-mass spectrometry (GC-MS). The fatty acids had a high phytochemical abundance at 73.8%, followed by tannins at 8.2%, carbohydrates at 6.9%, terpenoids at 4.3%, carboxylic acids at 2.5%, hydrocarbons at 2.4%, phenolic compounds at 0.2%, and sterols at 1.5%. Of these compounds, 22 were docked on the active side and on the catalytic dyad of His41 and Cys145 of the main protease of SARS-CoV-2 (Mpro). Eight active inhibitors were carbohydrates, five were fatty acids, three were terpenoids, two were carboxylic acids, one was a tannin, one was a phenolic compound, and one was a sterol. The best inhibitors were 4,8,13-Cyclotetradecatriene-1,3-diol, 1,5,9-trimethyl-12-(1-methylethyl), Andrographolide, and delta.4-Androstene-3.beta.,17.beta.-diol, with a binding affinity that ranged from -6.1 kcal/mol to -6.5 kcal/mol. The inhibitory effect of Saussurea costus of SARS-CoV-2 entry into the cell was studied using a pseudovirus with Spike proteins from the D614G variant and the VOC variants Gamma and Delta. Based on the viral cycle of SARS-CoV-2, our results suggest that the Saussurea costus aqueous extract has no virucidal effect and inhibits the virus in the events after cell entry. Furthermore, the biological activity of the aqueous extract was investigated against HSV-1 virus and two bacterial strains, namely Staphylococcus aureus ATCC BAA 1026 and Escherichia coli ATCC 9637. According to this study, an enormous number of water-soluble inhibitors were identified from Saussurea costus against the Mpro, and this is unprecedented as far as we know.

A study on the factors influencing the transfer of COVID-19 severe illness patients out of the ICU based on generalized linear mixed effect model.

The clinical data of 76 severe illness patients with novel coronavirus SARS-CoV-2 from July to August, 2020 admitted to the ICU Intensive Care Unit ward in a hospital in Urumqi were collected in the paper. By using the Laplace approximation parameter estimation method based on maximum likelihood estimation, the generalized linear mixed effect model (GLMM) was established to analyze the characteristics of clinical indicators in critical patients, and to screen the main influencing factors of COVID-19 critical patients' inability to be transferred out of the ICU in a short time: age, C-reactive protein, serum creatinine and lactate dehydrogenase.