The role of plasma serine leukocyte proteinase inhibitor in the body's defense against COVID-19

The COVID-19 pandemic continues, causing colossal damage to the population and the global economy. As COVID-19 is studied, new data are emerging regarding the risk of severe coronavirus infection in patients with α1-antitrypsin deficiency. α1 -Antitrypsin is the main inhibitor and key endogenous regulator of the serine leukocyte proteinase activitry released from the granules of activated neutrophils to the cell surface and into the extracellular space. It has been established that the number of cases of severe course and death of COVID-19 in the territories of 68 countries of the world correlates with the frequency of the spread of mutations in the proteinase inhibitor gene among the population of these countries, at which the concentration of α1-antitrypsin in the human blood plasma is 10 times lower than normal. All this contributes to the revision of a number of provisions of the pathogenesis and therapy of a new coronavirus infection.The review presents an analysis of the literature on the role of an inhibitor of serine leukocyte proteinases in protecting the body from COVID-19. The participation of α1-antitrypsin in the inhibition of SARS-CoV-2 penetration into the respiratory tract epithelial cells, in the protection of the vascular endothelium, blood plasma proteins and elastin of the lung tissue from the damaging effect of leukocyte elastase released during neutrophil degranulation and the formation of neutrophil extracellular traps (NETs) is considered. The role of a1-antitrypsin in suppressing inflammation by limiting the secretion of proinflammatory cytokines and neutrophil extracellular traps into the blood has been shown. The individual links in the pathogenesis of the new coronavirus infection have been detailed, which will allow revising the strategy for reducing the risks of severe course of COVID-19.

Immunological Indicators of Infl ammation in Late-Life Bipolar Disorder

The aim of the study was to determine the immune markers of infl ammation in the blood plasma of the elderly patients with bipolar affective disorders (BD) in relation to the clinical specifi cities of the disease. Patients and methods: 134 blood samples from the elderly patients aged from 52 to 88 years old (mean age 66.7 ± 7.7 years) with diagnose of bipolar disorder were examined. Infl ammatory markers in the blood plasma were determined as follows: the enzymatic activity of leukocyte elastase (LE) and the functional activity of the D1-proteinase inhibitor (D1-PI), as well as the level of autoantibodies (aAB) to S100b and myelin basic protein (MBP), and the protease inhibitor index (PII), which was the ratio of LE and D1-PI activity and characterized the activity of the proteolytic system as the most important component of infl ammation. Cluster analysis was used to reveal immunotypes. Results and discussion: а signifi cant increase in D1-PI and the level of aAB to S100b was revealed in elderly patients diagnosed with bipolar disorders, as well as low proteolytic activity of infl ammation (according to PII). Immune markers of infl ammation in different types of affective episodes (depressions, manias, mixed affective states) and in therapeutic remission did not differ from each other. Immunological parameters in elderly patients with bipolar disorders depended on the severity degree of the affective disorder. A relationship was found between the severity of depression and the level of aAB to S100b; the difference between mania and hypomania in terms of LE and PII activity was shown; in mixed affective states immunological parameters differed from the control only in moderate disorders. Remission with residual symptoms differed from asymptomatic therapeutic remission in terms of LE and PII activity. The two identifi ed clusters (immunotypes) differed in the activity of LE and PII. Conclusion: the results indicated the participation of infl ammation in the pathogenesis of bipolar disorder, and the isolated immunotypes confi rmed the clinical diversity of the disease. The study of the pathogenetic signifi cance of infl ammation and the identifi cation of various immunotypes was aimed at search for new therapy targets, taking into account the contribution of infl ammation.

Cluster analysis of blood serum inflammation markers of conditionally healthy people

Determination of inflammatory markers in blood of conventionally healthy people is of interest due to opportunity of detecting diseases at early (preclinical) stages, as well as latent forms of pathological processes. The level of inflammation may serve as an additional criterion to forming control groups in clinical and biological studies. The aim of the study is to identify some inflammatory and autoimmune markers in a group of conventionally healthy people and to conduct a cluster analysis of the data obtained. The study involved 100 apparently healthy people (without clinical signs of infections, somatic, neurological or mental diseases) aged 19 to 88 years. The levels of IL-10, TNFα, IL-6 and autoantibodies to S100b and MBP were determined in blood serum using ELISA. Enzymatic activity of leukocyte elastase (LE) and functional activity of the α1-proteinase inhibitor (α1-PI) were determined spectrophotometrically. Protease inhibitory index (PII) was calculated as the ratio of LE to α1-PI. Cluster analysis, as well as the Shapiro–Wilk, Kruskal–Wallis, and ANOVA methods were used as the main approach to statistical data processing. All the subjects were divided into three clusters, according to their immunological parameters. The selected clusters were statistically significantly different from each other, in terms of LE activity, protease-inhibitory index (PII), as well as IL-10 and TNFα levels. The indices of a distinct cluster (43% of total cohort) are most close to average indices assessed for the general sample, which gives ground to consider the values of immune indicators from this cluster as physiological norm, corresponding to the background immunity state in healthy people. Combination of immunological parameters in two other clusters (30 and 27% of the subjects, respectively) may reflect different variants of inflammatory reactions. These clusters are characterized by multidirectional changes in LE activity and protease-inhibitory index, compared to the standard values, thus suggestive for different variants of latent inflammatory reactivity, which are realized in the patients presented in these clusters. The obtained clusters did not differ by age of the subjects (p = 0.3476), which makes it possible to exclude a significant influence of age on the determined immune parameters, and by gender characteristics (p = 0.7233). The selected clusters did not differ statistically in the functional activity of α1-PI and in the level of autoantibodies to S100b and MBP. Thus, the group of conditionally healthy people is heterogeneous in terms of inflammation markers. Inflammatory reactions of varying severity were detected in about half of the cases. Probably, this may indicate the presence of a latent pathological process and requires a detailed clinical examination.

Proteomics-Based Identification of Salivary Changes in Patients with Burning Mouth Syndrome

Burning mouth syndrome (BMS) is a chronic oral condition characterized by an intraoral burning sensation, taste alterations, and dry mouth sensations. Although a number of factors have been closely related to the appearance of the symptoms, including anxiety, depression, and sleep disturbances, the etiology of BMS remains unclear. Furthermore, currently no objective diagnostic tools exist, making its diagnosis challenging. Therefore, to contribute to the knowledge about BMS etiology and look for objective tools for its diagnosis, the present study was conducted. Thus, the aim of this study was to analyze the proteomic profile of the resting whole saliva of patients with BMS and age and sex-matched controls using two-dimensional electrophoresis. The results showed evidence of changes in saliva at the level of proteins related to important pathways such as stress (sAA), immune system (Ig), and inflammation (leukocyte elastase inhibitor). While some of our findings have been previously described others, such as the deregulation of the coiled-coin domain containing protein 25 in BMS, are presented here for the first time to our knowledge. Thus, saliva provides us with relevant information about BMS pathophysiology and could be considered a suitable biofluid for its study and/or diagnosis.

Lossen Rearrangement of p-Toluenesulfonates of N-Oxyimides in Basic Condition, Theoretical Study, and Molecular Docking

The sulfonic esters of N-oxyimides are a group of compounds with a wide range of biological activities, as well as a unique reactivity toward amines. They undergo this reaction with primary amines and other nucleophilic reagents according to a Lossen-like rearrangement. The reaction is initiated by nucleophilic attack on a carbonyl group in the succinimide ring followed by isocyanate formation, which next interacts with another nucleophile molecule forming an addition product (e.g., ureido or urethane derivative). However, the secondary amines are also susceptible to other reactions leading to products containing the maleimide ring formed by sulphonic acid elimination. In the case of tertiary amines, this reaction is predominant. To explain the phenomenon of the reactivity of the N- oxyimides toward different types of amines, we employed various spectroscopic and X-ray approaches as well as DFT calculation. Results suggest that the basicity of the amine used for the reaction plays a crucial role in the reaction mechanism that eventually dominates the entire chemical process. Moreover, we applied molecular docking to investigate the ability of the products to act as serine protease inhibitors using human leukocyte elastase (HLE).

Premature Birth Infants Present Elevated Inflammatory Markers in the Meconium

Introduction: Prematurity, a well-established risk factor for various intestinal diseases in newborns, results in increased morbidity and mortality. However, the intestinal inflammatory status of preterm (PT) infants has been poorly characterized. Here we have broadly described the intestinal and systemic inflammatory status of PT children.Materials and Methods: Meconium and plasma from 39 PT and 32 full term (T) newborns were studied. Fecal calprotectin, polymorphonuclear leukocyte elastase (PMN-E), TNF, IL-17A, IL-8, IP-10, MCP-1, MIP-1, IL-1β, IL-1α, and E-selectin and the enzymatic activities of myeloperoxidase (MPO) and alkaline phosphatase (AP) in meconium were measured. Plasma levels of AP, hepatocyte growth factor, nerve growth factor (NGF), proinflammatory cytokines, leptin, adiponectin, PAI-1, and resistin were also determined. Correlations with gestational age (GA) and birth weight (BW) were studied.Results: Neutrophil derived PMN-E, MPO and calprotectin were increased in the meconium of PT compared to T newborns, while AP was decreased. No significant differences were found in other inflammatory parameters. Considering data from all children, GA and BW showed inverse correlation with neutrophil markers, while AP directly correlated with BW. Plasma levels of IL-1β and NGF were enhanced in PT infants, and were also negatively correlated with BW. PT children additionally showed neutropenia and decreased adiponectin, leptin, haematocrit, and haemoglobin. These parameters (neutrophils, adiponectin, and so forth) were positively correlated with GA and BW, while IL-8, MCP-1, PAI-1, and plasma AP were negatively correlated. PT children showing postnatal morbidity exhibited increased meconium MPO and MIP-1α.Conclusion: PT neonates present a significant elevation of intestinal inflammatory parameters, characterized by the presence of neutrophil markers, associated with mild systemic inflammation.

The origin and evolution of antistasin-like proteins in leeches (Hirudinida, Clitellata)

Bloodfeeding is employed by many parasitic animals and requires specific innovations for efficient feeding. Some of these innovations are molecular features that are related to the inhibition of hemostasis. For example, bloodfeeding insects, bats and leeches release proteins with anticoagulatory activity through their salivary secretions. The antistasin-like protein family, composed of serine protease inhibitors with one or more antistasin-like domains, is tightly linked to inhibition of hemostasis in leeches. However, this protein family has been recorded also in non-bloodfeeding invertebrates, such as cnidarians, molluscs, polychaetes and oligochaetes. The present study aims to: (1) root the antistasin-like gene tree and delimit the major orthologous groups, (2) identify potential independent origins of salivary proteins secreted by leeches and (3) identify major changes in domain and/or motif structure within each orthologous group. Five clades containing leech antistasin-like proteins are distinguishable through rigorous phylogenetic analyses based on nine new transcriptomes and a diverse set of comparative data: the trypsin + leukocyte elastase inhibitors clade, the antistasin clade, the therostasin clade and two additional, unnamed clades. The antistasin-like gene tree supports multiple origins of leech antistasin-like proteins due to the presence of both leech and non-leech sequences in one of the unnamed clades, but a single origin of factor Xa and trypsin + leukocyte elastase inhibitors. This is further supported by three sequence motifs that are exclusive to antistasins, the trypsin + leukocyte elastase inhibitor clade and the therostasin clade, respectively. We discuss the implications of our findings for the evolution of this diverse family of leech anticoagulants.

Feedlot diets containing increasing starch levels and different feed additives changes cecal proteome profile involved on energy metabolism and inflammatory response of Nellore cattle

Background Diets for feedlot cattle require higher energy density, thus contributing to the high rate of fermentable carbohydrate. The use of feed additives is necessary to reduce possible metabolic disorders. The objective of this study was to analyze the post-rumen effects of different levels of starch (25, 35, and 45%) and additives (Monensin, Blend of essential oil + exogenous α-Amylase) in diets for Nellore cattle feedlot. The cecum tissue proteome was separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), and then, with the verification of differentially expressed protein SPOTS, these were characterized by electrospray ionization mass spectrometry (ESI-MS/MS).Results The expression of nine enzymes participating in the Steps of the glycolysis pathway was verified, such as: Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), Glyceraldehyde-3-phosphate dehydrogenase like-17 protein, Triosephosphate isomerase (Step 1); Phosphoglycerate mutase and Phosphoglycerate mutase 2 (Step 2); Alpha-enolase (ENO1), Beta-enolase (ENO3) and Fructose-bisphosphate aldolase (ALDOB) (Step 4); and Pyruvate Kinase (PKM) (Step 5). There was expression of three enzymes linked to catalytic activities participating in the synthesis of lactate from pyruvate: L-lactate dehydrogenase B, L-lactate dehydrogenase A chain and L-lactate dehydrogenase. The ATP synthase subunit beta and ATP synthase subunit beta_mitochondrial participate in the electron transport chain, producing ATP from ADP in the presence of a proton gradient across the membrane. Due to the manipulation of diets, the expression of the Leukocyte elastase inhibitor protein, associated with the inflammatory response.Conclusions The use of blends of essential oil associated with α-amylase as a feed additive promoted the greater expression of enzymes in the pathway of glycolysis and gluconeogenesis (and the absence of proteins linked to inflammation (Leukocyte elastase inhibitor) in cecum tissues. On the other hand, the increase in starch in the diets promoted a reduction in enzymes linked to carbohydrate degradation with increased responses linked to inflammatory injuries.

Evidence-Based Psychiatry: Paraclinical Diagnostics of Asthenic Syndrome in Schizophrenia Based on the Determination of Leukocyte-Inhibitory Index

A leukocyte-inhibitory index (LII) is the ratio of the proteolytic enzyme leukocyte elastase (LE) to its inhibitor, an α1- proteinase inhibitor (α1-PI). LII characterizes the activity of the proteolytic system and can be considered as a potential objective criterion that determines both the course and the outcome of the disease. The changes of LII in schizophrenia patients with clinically diagnosed asthenia (schizoasthenia) and patients with schizophrenia without clinical signs of this syndrome were revealed. The objective: to study the possibility of the 95% confidence intervals for a comparative assessment of LII in patients with schizoasthenia and patients with schizophrenia without clinical signs of asthenic syndrome to obtain correct statistical conclusions. Patients and methods: Overall, 95 patients aged 20–55 years with paroxysmal-progressive (F20.x1) and paranoid (F20.00) schizophrenia were examined: 61 patients in the total sample were clinically diagnosed with asthenic symptom-complex. The enzymatic activity of LE and the functional activity of α1-PI were determined in blood serum. LII was calculated according to the formula. The confidence intervals were built using 4 different methods: Fieller’s theorem, delta method, regression methods and bootstrap method. Results: the statistical analysis indicates that the 95% confidence intervals of these indicators for the examined patient groups do not overlap. Therefore, these indicators relate to different populations, which mean the examined groups are characterized by different variants of the ratio of the proteolytic system components. Conclusion: the assessment of LII can serve as an objective statistically correct criterion for presence or absence of asthenic disorder in patients with schizophrenia in addition to clinical examination.