Glutamine Deficiency Promotes Immune and Endothelial Cell Dysfunction in COVID-19.

The coronavirus disease 2019 (COVID-19) pandemic has caused the death of almost 7 million people worldwide. While vaccinations and new antiviral drugs have greatly reduced the number of COVID-19 cases, there remains a need for additional therapeutic strategies to combat this deadly disease. Accumulating clinical data have discovered a deficiency of circulating glutamine in patients with COVID-19 that associates with disease severity. Glutamine is a semi-essential amino acid that is metabolized to a plethora of metabolites that serve as central modulators of immune and endothelial cell function. A majority of glutamine is metabolized to glutamate and ammonia by the mitochondrial enzyme glutaminase (GLS). Notably, GLS activity is upregulated in COVID-19, favoring the catabolism of glutamine. This disturbance in glutamine metabolism may provoke immune and endothelial cell dysfunction that contributes to the development of severe infection, inflammation, oxidative stress, vasospasm, and coagulopathy, which leads to vascular occlusion, multi-organ failure, and death. Strategies that restore the plasma concentration of glutamine, its metabolites, and/or its downstream effectors, in conjunction with antiviral drugs, represent a promising therapeutic approach that may restore immune and endothelial cell function and prevent the development of occlusive vascular disease in patients stricken with COVID-19.

New Short RNA Motifs Potentially Relevant in the SARS-CoV-2 Genome

Background: The coronavirus disease has led to an exhaustive exploration of the SARS-CoV-2 genome. Despite the amount of information accumulated, the prediction of short RNA motifs encoding peptides mediating protein-protein or protein-drug interactions has received limited attention. Objective(s): The study aims to predict short RNA motifs that are interspersed in the SARS-CoV-2 genome. Method(s): A method in which 14 trinucleotide families, each characterized by being composed of triplets with identical nucleotides in all possible configurations, was used to find short peptides with biological relevance. The novelty of the approach lies in using these families to search how they are distributed across genomes of different CoV genera and then to compare the distributions of these families with each other. Result(s): We identified distributions of trinucleotide families in different CoV genera and also how they are related, using a selection criterion that identified short RNA motifs. The motifs were reported to be conserved in SARS-CoVs;in the remaining CoV genomes analysed, motifs contained, exclusively, different configurations of the trinucleotides A, T, G and A, C, G. Eighty-eight short RNA motifs, ranging in length from 12 to 49 nucleotides, were found: 50 motifs in the 1a polyprotein-encoding orf, 27 in the 1b polyprotein-encoding orf, 5 in the spike-encoding orf, and 6 in the nucleocapsid-encoding orf. Although some motifs (~27%) were found to be intercalated or attached to functional peptides, most of them have not yet been associated with any known functions. Conclusion(s): Some of the trinucleotide family distributions in different CoV genera are not random;they are present in short peptides that, in many cases, are intercalated or attached to functional sites of the proteome.Copyright © 2022 Bentham Science Publishers.

Biomolecules of the Horseshoe Crab's Hemolymph: Components of an Ancient Defensive Mechanism and Its Impact on the Pharmaceutical and Biomedical Industry

Without adaptive immunity, invertebrates have evolved innate immune systems that react to antigens on the surfaces of pathogens. These defense mechanisms are included in horseshoe crab hemocytes' cellular responses to pathogens. Secretory granules, large (L) and small (S), are found on hemocytes. Once the invasion of pathogens is present, these granules release their contents through exocytosis. Recent data in biochemistry and immunology on the granular constituents of granule-specific proteins are stored in large and small granules which are involved in the cell-mediated immune response. L-granules contain most clotting proteins, which are necessary for hemolymph coagulation. They also include tachylectins;protease inhibitors, such as cystatin and serpins;and anti-lipopolysaccharide (LPS) factors, which bind to LPS and agglutinate bacteria. Big defensin, tachycitin, tachystatin, and tachyplesins are some of the essential cysteine-rich proteins in S-granules. These granules also contain tachycitin and tachystatins, which can agglutinate bacteria. These proteins in granules and hemolymph act synergistically to fight infections. These biomolecules are antimicrobial and antibacterial, enabling them to be drug resistant. This review is aimed at explaining the biomolecules identified in the horseshoe crab's hemolymph and their application scopes in the pharmaceutical and biotechnology sectors.Copyright © 2022 Md. Ashrafuzzaman et al.

Study in-silico oleanane triterpenoids in Aquilaria spp. as a COVID-19 antiviral

Covid-19, a disease characterized by Severe Acute Respiratory Syndrome, is caused by Coronavirus-2 (SARS-CoV-2). This virus causes tissue damage and a decrease in the respiratory system. Agarwood (Aquilaria spp) is a plant that has various pharmacological activities, including relieving respiratory diseases. One of the several secondary metabolites reported in Aquilaria spp. is oleanane triterpenoids, suspected of having antiviral activity. This research was aimed to determine the potential of oleanane triterpenoids from Agarwood as a covid-19 antiviral by in silico study. The research methods were molecular docking, prediction of Lipinski rules of five, and prediction of ADME. As a receptor, main protease (Mpro) Covid-19 was used. The four oleanane triterpenoid compounds in Agarwood demonstrated a higher affinity for the main protease covid-19 (G 11-oxo-beta-amyrin = -9.8 kcal/mol, G hederagenin-an = -9.6 kcal/mol, G 3beta-acetoxyfriedelane = -9.4 kcal/mol, G ursolic acid = -9.5 kcal/mol) than Lopinavir (G = -6.2 kcal/mol) and Remdesivir (G = -7.2 kcal/mol). The major amino acids involved in ligand and receptor interactions are methionine 49 and 165, proline 168, glutamine 189, arginine 188, and threonine 25. According to the prediction of Lipinski's rule of five and ADME, hederageninan is potential for development as oral medicine.

COVID-19 Metabolomic-Guided Amino Acid Therapy Protects from Inflammation and Disease Sequelae.

The outbreak of coronavirus disease 2019 (COVID-19) has caused a worldwide pandemic since 2019. A metabolic disorder is a contributing factor to deaths from COVID-19. However, the underlying mechanism of metabolic dysfunction in COVID-19 patients and the potential interventions are not elucidated. Here targeted plasma metabolomic is performed, and the metabolite profiles among healthy controls, and asymptomatic, moderate, and severe COVID-19 patients are compared. Among the altered metabolites, arachidonic acid and linolenic acid pathway metabolites are profoundly up-regulated in COVID-19 patients. Arginine biosynthesis, alanine, aspartate, and glutamate metabolism pathways are significantly disturbed in asymptomatic patients. In the comparison of metabolite variances among the groups, higher levels of l-citrulline and l-glutamine are found in asymptomatic carriers and moderate or severe patients at the remission stage. Furthermore, l-citrulline and l-glutamine combination therapy is demonstrated to effectively protect mice from coronavirus infection and endotoxin-induced sepsis, and is observed to efficiently prevent the occurrence of pulmonary fibrosis and central nervous system damage. Collectively, the data reveal the metabolite profile of asymptomatic COVID-19 patients and propose a potential strategy for COVID-19 treatment.

Protein losing enteropathy (PLE) secondary to norovirus infection in a 4 year old patient

Case Report: Protein losing enteropathy (PLE) occurs when proteins leak from the gastrointestinal (GI) system more rapidly than they are produced. Inflammation of the GI tract facilitates increased membrane permeability of gastric mucosa, leading to excess protein leakage. 1 PLE in children has been associated with CMV, rotavirus, COVID-19, HIV, C. difficile, and autoimmune diseases like Crohn's Disease. 2-6 Norovirus is a known cause of PLE in immunocompromised pediatric patients. 7-8 However, to our knowledge, there are no case reports about PLE precipitated by norovirus in immunocompetent pediatric patients. The purpose of this case report is to present a case of PLE precipitated by a norovirus infection in a 4- year-old previously healthy child. While the above gastrointestinal viruses have been proposed as precipitators for this disease, PLE precipitated by norovirus infection has not been well described. This case also highlights the importance of early diagnosis and management to avoid complications. Method(s): Our patient initially presented with two days of abdominal pain, diarrhea, emesis, reduced urine output, and swelling of the lower extremities. He was exposed to several sick family members-his sister had upper respiratory symptoms and his grandmother had gastrointestinal symptoms. Physical exam was notable for diminished breath sounds in the right lower lobe, abdominal distension with diffuse tenderness and dullness to percussion, significant scrotal and penile edema, and bilateral lower extremity pitting edema. Laboratory results revealed leukocytosis, hypoalbuminemia, hyponatremia, elevated aspartate aminotransferase (AST), and elevated serum alpha-1-antitrypsin, as well as low Immunoglobulins G and M. CD3 and CD4 levels were low reflecting cellular immune dysregulation seen in patients with PLE. IgA and Tissue Transglutaminase (TTF) were within normal limits. Ebstein Barr Virus and cytomegalovirus IgM antibodies were negative. COVID IgG was negative as well. His Polymerase chain reaction (PCR) gastrointestinal panel was positive for norovirus. A chest X-ray showed a large right pleural effusion. Abdominal CT revealed large ascites slightly more predominant in the upper abdomen, mesenteric lymphadenitis, and bilateral pleural effusions. Echocardiogram showed small anterior and apical pericardial effusions. Result(s): Based on the patient's elevated serum alpha-1 antitrypsin levels, hypoalbuminemia, low levels of immunoglobulins and lymphocytes, and clinical manifestations of ascites, bilateral pleural effusions, pericardial effusion, and dependent edema, along with a positive PCR for norovirus, the diagnosis of PLE secondary to Norovirus was made. Conclusion(s): This case demonstrates the importance of recognizing viruses like Norovirus as potential causes of PLE to avoid a delay in diagnosis and initiation of therapy, and to avoid unnecessary additional testing. Copyright © 2023 Southern Society for Clinical Investigation.

Dysregulation of glutamine/glutamate metabolism in COVID-19 patients: A metabolism study in African population and mini meta-analysis.

Coronavirus disease 2019 (COVID-19) remains a serious global threat. The metabolic analysis had been successfully applied in the efforts to uncover the pathological mechanisms and biomarkers of disease severity. Here we performed a quasi-targeted metabolomic analysis on 56 COVID-19 patients from Sierra Leone in western Africa, revealing the metabolomic profiles and the association with disease severity, which was confirmed by the targeted metabolomic analysis of 19 pairs of COVID-19 patients. A meta-analysis was performed on published metabolic data of COVID-19 to verify our findings. Of the 596 identified metabolites, 58 showed significant differences between severe and nonsevere groups. The pathway enrichment of these differential metabolites revealed glutamine and glutamate metabolism as the most significant metabolic pathway (Impact = 0.5; -log10P = 1.959). Further targeted metabolic analysis revealed six metabolites with significant intergroup differences, with glutamine/glutamate ratio significantly associated with severe disease, negatively correlated with 10 clinical parameters and positively correlated with SPO2 (rs = 0.442, p = 0.005). Mini meta-analysis indicated elevated glutamate was related to increased risk of COVID-19 infection (pooled odd ratio [OR] = 2.02; 95% confidence interval [CI]: 1.17-3.50) and severe COVID-19 (pooled OR = 2.28; 95% CI: 1.14-4.56). In contrast, elevated glutamine related to decreased risk of infection and severe COVID-19, the pooled OR were 0.30 (95% CI: 0.20-0.44), and 0.44 (95% CI: 0.19-0.98), respectively. Glutamine and glutamate metabolism are associated with COVID-19 severity in multiple populations, which might confer potential therapeutic target of COVID-19, especially for severe patients.

Glutamine-Driven Metabolic Adaptation to COVID-19 Infection.

Background: COVID-19 is known to be transmitted by direct contact, droplets or feces/orally. There are many factors which determines the clinical progression of the disease. Aminoacid disturbance in viral disease is shown in many studies. In this study we aimed to evaluate the change of aminoacid metabolism especially the aspartate, glutamine and glycine levels which have been associated with an immune defence effect in viral disease. Methods: Blood samples from 35 volunteer patients with COVID-19, concretized diagnosis was made by oropharyngeal from nazofaringeal swab specimens and reverse transcriptase-polymerase chain reaction, and 35 control group were analyzed. The amino acid levels were measured with liquid chromatography-mass spectrometry technology. Two groups were compared by Kolmogorov-Smirnov analysis, Kruskal-Wallis and the Mann-Whitney U. The square test was used to evaluate the tests obtained by counting, and the error level was taken as 0.05. Results: The average age of the patient and control group were 48.5 ± 14.9 and 48.8 ± 14.6 years respectively. The decrease in aspartate (p = 5.5 × 10-9) and glutamine levels (p = 9.0 × 10-17) were significiantly in COVID group, whereas Glycine (p = 0.243) increase was not significiant. Conclusions: Metabolic pathways, are affected in rapidly dividing cells in viral diseases which are important for immun defence. We determined that aspartate, glutamine and glycine levels in Covid 19 patients were affected by the warburg effect, malate aspartate shuttle, glutaminolysis and pentose phosphate pathway. Enteral or parenteral administration of these plasma amino acid levels will correct the duration and pathophysiology of the patients' stay in hospital and intensive care.

State of Pediatric Celiac Disease Practice in North America: A Naspghan 2021 Survey

Background and Aims: Substantial practice variation exists in both the diagnostic criteria for and the post-diagnosis monitoring of celiac disease (CeD). Differences include standards for serological diagnosis, endoscopic practices, models of care, and long-term clinical monitoring, all confounded by the COVID-19 pandemic. With the exponential rise of gluten-related disorders, revised ESPGHAN guidelines and new healthcare barriers, it is helpful to explore practice patterns to inform updates to clinical guidelines and future research endeavors. The purpose of this survey was to understand the expertise and practice parameters of pediatric gastroenterology (GI) clinicians across North America for the diagnosis and management of children with celiac disease. Method(s): A 23-item survey designed by a working committee of the NASPGHAN Celiac Disease Special Interest Group was distributed electronically to NASPGHAN members, including attending physicians, fellows, and advanced practice providers from September to December 2021. Four themes were explored: 1) screening and diagnosis;2) monitoring;3) impact of the COVID-19 pandemic;and 4) education and training. The implementaion of the ESPGHAN non-biopsy serologic diagnosis (based on the 2020 guidelines: tissue transglutaminase IgA (TTG-IgA) 10x upper limit of normal and a second sample with a positive endomysial antibody) by providers was explored. Descriptive statistics were tabulated by region, clinical role and those who identified as working at a celiac center. Result(s): A total of 284 surveys were completed with a response rate of 11.1% (264/2552). The majority of respondents were from the United States (89%, n=235) and Canada (8%, n=22) with 2% (n=5) from Mexico. Serology-based diagnosis as per ESPGHAN 2020 guidelines was accepted by 54.5% (n=12/22) of Canadian respondents and 39.6% (n=93/235) from the U.S (p=0.17). Since the COVID-19 pandemic, 36% of respondents have increased their application of non-biopsy diagnosis. Canadian respondents reported offering the ESPGHAN non-biopsy approach to diagnosis more often during the COVID-19 pandemic (Canada 74% vs US 33%, <0.0001). A higher precentage of patients who lived in Canada (52%) with positive celiac serologies waited >1 month to be evaluated by GI than the US (30%);p=0.03. There was also a significant difference between access to endoscopy within a month between patients who lived in Canada and the US patients (Canada 77% >1 month, US 20% >1 month;p=<0.001). Investigations at follow-up which were completed most frequently by those who identified as working at a celiac center (n=108) included complete blood count, thyroid function tests, liver enzymes, iron profile, Vitamin D and TTG-IgA (Figure 1). Among these respondents, 49.1% (n=53/108) repeat family screening ranging every 1-5 years. Specialty training in CeD remains limited as only 25.7% (n=61/237) staff pediatric gastroenterologists had celiac-focused didactic lectures, and 23.3% (n=55/237) participated in a CeD specialty clinic during their fellowship. Conclusion(s): This survey revealed heterogeneity in current practices for the diagnosis and management of CeD in North America and the influence of the COVID-19 pandemic in increasing the use of the ESPGHAN no-biopsy approach to diagnosis. An education gap was identified for CeD in pediatric GI fellowship training. Further studies are needed to understand the impact of these variable practices and future research priorities and clinical guidelines should take this variation into consideration. (Figure Presented).

Celiac Disease Screening and Autoimmunity in the Era of the Covid-19 Pandemic

Celiac disease (CD) is a common immune-mediated disease of the gastrointestinal tract that results in small bowel damage from ingesting gluten in genetically susceptible patients. The SARS-CoV-2 (COVID-19) pandemic resulted in a nationwide healthcare lockdown, with the halting of elective medical procedures and tests. The potential consequence of the COVID-19 pandemic on the long-term risk of systemic and autoimmune disorders like CD is not yet clear. There is growing evidence that COVID-19 can cause a "cytokine storm" affecting epithelial cells like intestinal mucosal cells, which may increase the risk of celiac autoimmunity;defined as positive celiac serology like anti-tissue transglutaminase immunoglobulin A (TTG IgA). Many providers report observing an increased number of newly diagnosed autoimmune disorders such as the CD post-COVID-19 pandemic. It is not clear if this is a true increase in CD autoimmunity or due to overcompensation of diagnostic numbers after the lockdown. This study aims to evaluate the effect of the COVID-19 pandemic on the frequency of CD screening tests and the rate of celiac autoimmunity reflected by positive TTG IgA titers. Method(s): This retrospective cohort study used the Rochester Epidemiology Project (REP) electronic health records between January 2018 and December 2021. All subjects who underwent celiac serology screening testing (TTG IgA) during the study period were included. The REP 7-county core region includes Olmsted County and the following 6 counties: Wabasha, Waseca, Steele, Dodge, Freeborn, and Mower. We documented the total number of celiac serology screening (TTG IgA) tests ordered during the study period and compared the frequency of tests performed and the rate of positivity before and after COVID-19. We used January 2020 as a marker of the start of the COVID- 19 pandemic in the USA;referring to the first reported case by the CDC. All patients had active research authorization at the onset of the study before being included in the analysis. All data were stored and managed in compliance with the REP and Mayo Clinic IRB regulations. Result(s): A total of 15,443 (Mean=321.7, SD= 6.7, CI= 13.4) TTG IgA screening tests for CD were ordered from January 2018 till December 2021. As expected, the frequency of celiac screening tests ordered decreased sharply during the lockdown in early 2020 (March-May) (Mean= 202.7, SD=39.8, CI= 171.4)), but it picked rapidly by June 2020 with a 2.5% increase to the pre-pandemic testing frequency rate (Figure1). Despite this sharp decrease in celiac screening testing during the lockdown, the rate of celiac autoimmunity (positive TTG IgA) was higher during similar periods in pre and post-lock down years Figure1. Comparing the rate of positivity of celiac tests done during the months of March to May of the lockdown in 2020 (44/608;7.24%) to the same period during years 2018, 2019, and 2021, we found a statistical difference in the rate of positivity (p= 0.04, df= 3) figure2. There was no significant difference in the number of tests administered between pre-and post-COVID years (7625 vs. 7818, p = 0.09). Over the study period, the total number of positive screening tests was 655/15443 (4%). Using January 2020 as our cut-off, we compared the rate of screening test positivity pre and post COVID-19 to the total number of tests as demonstrated in figure1. Conclusion(s): Celiac screening and autoimmunity were stable over the study period, however, during the lockdown, the rate of celiac autoimmunity was significantly higher as compared to similar periods before and after the pandemic year. It is not clear if this is due to false-positive TTG IgA post-COVID, or true autoimmunity. We expect that follow-up studies with a longer follow-up period during and after the COVID-19 pandemic are needed to understand the true effect of COVID-19 on celiac autoimmunity. (Figure Presented).

SARS-CoV-2 infection impacts carbon metabolism and depends on glutamine for replication in Syrian hamster astrocytes.

COVID-19 causes more than million deaths worldwide. Although much is understood about the immunopathogenesis of the lung disease, a lot remains to be known on the neurological impact of COVID-19. Here, we evaluated immunometabolic changes using astrocytes in vitro and dissected brain areas of SARS-CoV-2 infected Syrian hamsters. We show that SARS-CoV-2 alters proteins of carbon metabolism, glycolysis, and synaptic transmission, many of which are altered in neurological diseases. Real-time respirometry evidenced hyperactivation of glycolysis, further confirmed by metabolomics, with intense consumption of glucose, pyruvate, glutamine, and alpha ketoglutarate. Consistent with glutamine reduction, the blockade of glutaminolysis impaired viral replication and inflammatory response in vitro. SARS-CoV-2 was detected in vivo in hippocampus, cortex, and olfactory bulb of intranasally infected animals. Our data evidence an imbalance in important metabolic molecules and neurotransmitters in infected astrocytes. We suggest this may correlate with the neurological impairment observed during COVID-19, as memory loss, confusion, and cognitive impairment.

Combined exploration of the mechanism of Sang Xing Decoction in the treatment of smoke-induced acute bronchitis from protein and metabolic levels.

Sang Xing decoction (SXD) is a typical prescription for treating "warm dryness" in traditional Chinese medicine (TCM), which is equivalent to respiratory diseases such as acute bronchitis in modern medicine. However, its mechanism of action remains unclear. In this study, the representative components of SXD were characterized using liquid chromatography-tandem mass spectrometry (LC-MS). The key targets, signaling pathways, and metabolic pathways associated with SXD in the treatment of acute bronchitis were identified via network prediction and metabolomics. A rat model of acute bronchitis was also established using mixed smoke, systematic in vivo experiments such as histopathological analyses, enzyme-linked immunosorbent assay (ELISA), immunofluorescence, immunohistochemistry and western blotting were conducted to evaluate the network prediction results. An in-depth analysis of the targeted quantitative results was performed using the SIMCA software and MetaboAnalyst website. The results revealed that 50 active compounds and 45 key targets were screened and clustered with 20 approved drugs. The NF-κB signaling pathway, oxidative stress, and glutamine metabolism were associated with the therapeutic mechanism of SXD in acute bronchitis. In vivo experiments showed that SXD may maintain the production of inflammatory factors by regulating the PI3K/Akt/NF-κB signaling pathway, improving the metabolism of glutamine and glutamate to reduce oxidative stress, and inhibiting apoptosis. Simultaneously, the possibility of using SXD as an adjuvant drug for COVID-19 treatment was also revealed. This research will lay the foundation for the modern clinical application of SXD and promote the promotion and innovation of TCM.

MANAGEMENT OF SICKLE CELL DISEASE: A CLINICAL PRACTICE SURVEY

Background: Therapeutic options for Sickle Cell Disease (SCD) have increased recently as well as the development of updated national guidelines. It is not known how these options are being offered or to what degree guidelines are incorporated into clinical practice. Objectives: To assess the clinical practice patterns of providers treating children with SCD. Design/Method: A survey study was performed which included nine sections: clinic structure, prophylaxis, immunizations, hydroxyurea, splenic sequestration, stroke, novel therapies, potential curative therapies, and transition. Survey was disseminated over a three-month period via SurveyMonkey, to members of the American Society of Pediatric Hematology-Oncology Hemoglobinopathy Special Interest Group. Results: There were 86 respondents;most were attending/faculty (85%, 73/86) who were part of a university/academic practice (65%, 56/86). Program size was most commonly 50-250 patients (44%, 37/86). Accessibility to support staff in clinic included 95% (81/86) social work;76% (65/86) child life;68% (58/86) nurse coordinator and 34% (29/86) school liaison and 15% (13/86) transition navigator. For preventive care, 72% prescribe penicillin prophylaxis before 2 months of age recommending 100% (83) for HbSS and Sβnull, 72% (60/83) for HbSC and 70% (58/83) for HbSβplus. Influenza was the most common vaccine offered in clinic at 96% (76/79) with 91% (72/79) offering pneumococcal vaccines, 84% (67/79) offering meningococcal vaccines and 50% (40/79) offering COVID vaccines. Transcranial doppler screening was offered in 95% (69/73) but only 42% (31/73) performed MRI screening for silent stroke. Transfusion therapy was recommended for primary stroke prevention by 90% (65/72) and 84% (59/70) attempt to transition to hydroxyurea following TWITCH guidelines. For secondary stroke prevention, 88% (63/72) recommend chronic transfusion therapy. Regarding disease-modifying therapy, 90% (70/78) report starting hydroxyurea routinely in patients with HbSS and Sβnull;initiated at 9 months of age by 69% (54/78). Laboratory monitoring recommended every 3 months for stable dosing by 62% (49/78) and hydroxyurea held by 56% (44/78) if platelets <75,000, 73% (56/78) for neutrophils <1000. New therapies were recommended for patients on hydroxyurea who were still experiencing SCD complications: L-glutamine 68% (37/54;crizanlizumab 93% (54/58). Voxelotor was recommended for patients on hydroxyurea with low hemoglobin 65% (43/66). Matched sibling transplant was considered for any disease severity by 55% (38/69). Gene therapy trial is offered on-site by 29% (20/69). Transition programs were endorsed by 61% (42/69), but only 45% (31/68) had dedicated staff. Conclusion: This survey is the only assessment of the application of SCD guidelines in clinical practice.

MAIN BLOOD SERUM METABOLITES RESPONSIBLE FOR THE DISCRIMINATION OF HOSPITALIZED PATIENTS WITH MODERATE SYMPTOMS WITH OR WITHOUT COVID-19

Introduction: The variation in human blood serum metabolites resulting from an infection can assist in understanding mechanisms of pathogen action and body response and improve diagnosis. Aim: To map serum signatures of hospitalized symptomatic patients, positive or negative to SARS-CoV-2. Methods: Patients (n = 64) admitted to Anhembi Field Municipal Hospital, a hospital set up for initial care to patients with moderate symptoms, were analyzed being discriminated in positive (n = 32) or negative patients. Age and gender were matched to ensure homogeneity in the basal metabolic rates. Three Nuclear Magnetic Resonance (NMR) data set were recorded on Bruker AVANCE III 600 MHz spectrometer for serum samples analyzed in MetaboAnalyst 5.0 software platform. Results and discussion: The mean age of groups was 54.92 ±12.41 and 54.30 ±12.15, for positive and negative patients, divided in 16 female and 16 male. The ethnicity was 56.2% vs 46.8% caucasian, 34.3% mixed race in both groups, and 9.3% 12.5% vs black in positive and negative groups, respectively. BMI was 24 ±6.93 vs 33.5 ±7.85 in comparison to positive and negative patients, respectively. In both groups 50% of patients presented alveolar infiltrate. Although the groups were not paired by comorbidities, they were homogeneous ensuring that the metabolic variation is due to COVID-19 as similar percentage of patients with arterial hypertension, diabetes and dyslipidemia. Clinical symptoms were also remarkably similar between the groups in relation to: fever, dry cough, dyspnea and myalgia. The Partial Least Squares - Discriminant Analysis (PLS-DA) performed onto noesy1d data discriminated positively from negative patients. Also, it covered lower variance. Combining NMR techniques, it was possible to depict the main metabolites that distinguished the COVID-19 signatures. Alanine, glucose, cholesterol, and glutamine were increased, and lactate decreased in COVID-19. Conclusion: These results suggest NMR as an excellent tool to differentiate hospitalized patients with moderate symptoms as COVID-19 positive or negative. The Ethics Research Committee of the University of Campinas approved all of the experimental procedures, and all individuals signed the informed consent form.

[Innovations in peritoneal dialysis]. / Innovationen in der Peritonealdialyse.

Peritoneal dialysis used to be a common treatment for acute kidney failure that required dialysis. In favor of continuous, extracorporeal renal replacement procedures, it disappeared from the scene in the western world, whereas it continues to be used in structurally poor countries due to its simplicity and low resource intensity. Recently, the shortages in medical care in the context of the coronavirus disease 2019 (COVID-19) pandemic led to renewed worldwide interest in peritoneal dialysis as a safe option in acute kidney failure requiring dialysis. The introduction of biocompatible solutions 20 years ago was expected to reduce mortality or technical failure. Unfortunately, so far this could only be implied but not confirmed in studies. Immunomodulatory adjuvants are an innovative option which have the potential to improve the local immunocompetence and prevent the loss of peritoneal function. Currently, the vision of a wearable artificial kidney is getting closer. Intensification of dialysis dose also appears achievable with minimal dialysate volumes. In times of global warming, the regeneration of dialysates could not only save relevant amounts of water but also have a favorable impact on the CO2 balance. In summary, peritoneal dialysis is currently enjoying a comeback. This article describes the current and future developments of this procedure.

Activation of TCA cycle restrains virus-metabolic hijacking and viral replication in mouse hepatitis virus-infected cells.

BACKGROUND: One of coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has caused coronavirus disease 2019 (COVID-19) pandemic and threatened worldwide. However, therapy for COVID-19 has rarely been proven to possess specific efficacy. As the virus relies on host metabolism for its survival, several studies have reported metabolic intervention by SARS-CoV-2. RESULTS: We investigated the coronavirus-metabolic hijacking using mouse hepatitis virus (MHV) as a surrogate for SARS-CoV-2. Based on the altered host metabolism by MHV infection, an increase of glycolysis with low mitochondrial metabolism, we tried to investigate possible therapeutic molecules which increase the TCA cycle. Endogenous metabolites and metabolic regulators were introduced to restrain viral replication by metabolic intervention. We observed that cells deprived of cellular energy nutrition with low glycolysis strongly suppress viral replication. Furthermore, viral replication was also significantly suppressed by electron transport chain inhibitors which exhaust cellular energy. Apart from glycolysis and ETC, pyruvate supplement suppressed viral replication by the TCA cycle induction. As the non-glucose metabolite, fatty acids supplement decreased viral replication via the TCA cycle. Additionally, as a highly possible therapeutic metabolite, nicotinamide riboside (NR) supplement, which activates the TCA cycle by supplying NAD+, substantially suppressed viral replication. CONCLUSIONS: This study suggests that metabolite-mediated TCA cycle activation suppresses replication of coronavirus and suggests that NR might play a role as a novel therapeutic metabolite for coronavirus.

IMPACT OF A SARS-COV2 INFECTION IN PATIENTS WITH CELIAC DISEASE

Objective. No data are available on the effects of SARS-CoV2 infection in patients with celiac disease (CD) in terms of development of related symptoms and antibodies. We aimed to investigate the impact of SARS-CoV2 infection in CD. Design. During lockdown (March-May 2020), celiac patients living in Milano were interviewed about the development of COVID-19 resembling symptoms, adherence to an anti-virus lifestyle and gluten-free diet (GFD), and were asked to reply to a stress questionnaire. The development of anti SARSCoV2 IgG and IgA (anti RBD and N proteins) and the expression of duodenal ACE2 receptor were also investigated. Whenever available, duodenal histology, anti-tissue transglutaminase IgA (tTGA), immunologic comorbidities and GFD adherence were analyzed as possible risk factors. Results. 362 celiac patients have been interviewed and 42 (11%) reported COVID-19 resembling symptoms. The presence of symptoms was not influenced by positivity of tTGA, presence of duodenal atrophy or adherence to GFD. 37% of symptomatic patients showed anti SARS-CoV2 Immunoglobulins (Ig). Globally, 18% of celiac patients had anti SARS-CoV2 Ig vs 25% of non-celiac controls (p=0.18). Levels of anti RBD IgG/IgA and anti N IgG did not differ from non-celiac controls. Celiac patients had significantly lower levels of anti N IgA. ACE2 receptor was detected in the non-atrophic duodenal mucosa of celiac patients;atrophy was associated with a weaker expression of ACE2 receptor. Conclusion. CD patients show an anti SARS-CoV2 Ig positivity/profile similar to non-celiac controls, except for anti-N IgA. Main celiac biomarkers and adherence to the GFD do not influence the development of different antibody profiles.

Supplying the trip to antibody production-nutrients, signaling, and the programming of cellular metabolism in the mature B lineage.

The COVID pandemic has refreshed and expanded recognition of the vital role that sustained antibody (Ab) secretion plays in our immune defenses against microbes and of the importance of vaccines that elicit Ab protection against infection. With this backdrop, it is especially timely to review aspects of the molecular programming that govern how the cells that secrete Abs arise, persist, and meet the challenge of secreting vast amounts of these glycoproteins. Whereas plasmablasts and plasma cells (PCs) are the primary sources of secreted Abs, the process leading to the existence of these cell types starts with naive B lymphocytes that proliferate and differentiate toward several potential fates. At each step, cells reside in specific microenvironments in which they not only receive signals from cytokines and other cell surface receptors but also draw on the interstitium for nutrients. Nutrients in turn influence flux through intermediary metabolism and sensor enzymes that regulate gene transcription, translation, and metabolism. This review will focus on nutrient supply and how sensor mechanisms influence distinct cellular stages that lead to PCs and their adaptations as factories dedicated to Ab secretion. Salient findings of this group and others, sometimes exhibiting differences, will be summarized with regard to the journey to a distinctive metabolic program in PCs.

The effect of glutamine supplementation on serum levels of some inflammatory factors, oxidative stress, and appetite in COVID-19 patients: a case-control study.

BACKGROUND: Malnutrition is seen in COVID-19 patients, and reducing malnutrition with appropriate therapies may improve these patients' health. This case-control study aimed to assess and compare serum levels of some inflammatory factors, oxidative stress, and appetite in COVID-19 patients with respiratory infections that receive glutamine treatment with a control group. METHODS: In this study, patients who consented to use glutamine were considered as the case group and other patients who did not use glutamine were considered as a control group. Two hundred twenty-two COVID-19 patients (51.2 ± 6.7) using L-Glutamine and 230 COVID-19 patients (51.3 ± 8.2) with similar age, gender, and clinical status, as the control group, were included in the study. For 5 days, the case group consumed 10 g of glutamine supplement three times per day. At the end of the 5 days, blood samples were taken again to test for serum levels of IL1ß, tumor necrosis factor-α, malondialdehyde, and total antioxidant capacity, then all data were analyzed. RESULTS: Serum levels of ß-1 interleukin, tumor necrosis factor-α and hs-CRP were significantly reduced with five days of glutamine supplementation (p < 0.05), and patients' appetite during 5 days of glutamine supplementation compared with the control group had a significant increase (p < 0.05). CONCLUSION: Glutamine supplementation in COVID-19 patients with respiratory infection significantly reduces serum levels of interleukin-1 ß, hs-CRP, and tumor necrosis factor-α and significantly increases appetite, so glutamine supplementation may be useful for COVID-19 patients in the hospital.

Phytoconstituents from ten natural herbs as potent inhibitors of main protease enzyme of SARS-COV-2: In silico study.

Background: Lack of treatment of novel Coronavirus disease led to the search of specific antivirals that are capable to inhibit the replication of the virus. The plant kingdom has demonstrated to be an important source of new molecules with antiviral potential. Purpose: The present study aims to utilize various computational tools to identify the most eligible drug candidate that have capabilities to halt the replication of SARS-COV-2 virus by inhibiting Main protease (Mpro) enzyme. Methods: We have selected plants whose extracts have inhibitory potential against previously discovered coronaviruses. Their phytoconstituents were surveyed and a library of 100 molecules was prepared. Then, computational tools such as molecular docking, ADMET and molecular dynamic simulations were utilized to screen the compounds and evaluate them against Mpro enzyme. Results: All the phytoconstituents showed good binding affinities towards Mpro enzyme. Among them laurolitsine possesses the highest binding affinity i.e. -294.1533 kcal/mol. On ADMET analysis of best three ligands were simulated for 1.2 ns, then the stable ligand among them was further simulated for 20 ns. Results revealed that no conformational changes were observed in the laurolitsine w.r.t. protein residues and low RMSD value suggested that the Laurolitsine-protein complex was stable for 20 ns. Conclusion: Laurolitsine, an active constituent of roots of Lindera aggregata, was found to be having good ADMET profile and have capabilities to halt the activity of the enzyme. Therefore, this makes laurolitsine a good drug candidate for the treatment of COVID-19.