Peripheral Blood Of Individuals Research Articles

Molecular Characteristics of Blood Serum After Covid-19 Vaccination in a Remote Period

COVID-19 is a dangerous disease with long-lasting consequences. Vaccination contributes to the accumulation of neutralizing anti-S IgG antibodies, reducing the incidence of COVID-19 and its complications. However, in some individuals, the inflammatory process can persist for an indefinite period and lead to a wide range of dysfunctions. The current task is to investigate molecular markers for their detection. The aim of this study is to examine the levels of anti-S IgG antibodies, lactate, glucose, lactate dehydrogenase, and C-reactive protein in the peripheral blood of individuals who have and have not been affected by COVID-19 after vaccination. The research subject is venous blood. Among 547 employees of the Neurosurgery Institute (481 vaccinated against COVID-19 and 66 unvaccinated individuals), levels of anti-S IgG antibodies were investigated, as well as levels of lactate, lactate dehydrogenase, glucose, and C-reactive protein. At the time of the study, among 372 individuals, 16 months had passed from the first vaccination, and 12 months had passed from the second vaccination; in 21 individuals, 12 months had passed after a single vaccination, and in 88 individuals, 16 months had passed from the first vaccination, 12 months from the second, and 6 months from the third vaccination. Methods. Quantitative determination of IgG antibodies to the S protein of the SARS-CoV-2 virus. Confirmation of COVID-19 using the RT-PCR method (Allplex 2019-nCoV kit, SeeGene, Korea). Levels of lactate, lactate dehydrogenase, glucose, and C-reactive protein were determined using reagents from BioSystems (Spain). Statistical analysis of the obtained data was performed using Jamovi software (USA) and the following criteria: χ2 ‒ Kruskal-Wallis, W ‒ Dwass-Steel-Critchlow-Fligner (DSCF), χ2 ‒ Pearson, t ‒ Student, rs ‒ Spearman, τb ‒ Kendall. A statistically significant difference was considered at p < 0.05. Results. The level of anti-S IgG antibodies to the SARS-CoV-2 virus was higher in vaccinated individuals compared to unvaccinated individuals (Kruskal-Wallis χ2=14.09; p < 0.001). A higher level of antibodies to the S protein of the virus was observed when using the Comirnaty vaccine compared to vaccination with Moderna, AstraZeneca, Pfizer, and CoronaVac (Dwass-Steel-Critchlow-Fligner (DSCF): W 4.26, p=0.002; W 4.62, p=0.010; W 4.84, p=0.006, respectively). Vaccination reduces the likelihood of contracting the disease by 1.84 times (Odds Ratio (OR) 1.84; 95% Confidence Interval (CI) 1.02‒3.30; χ2=4.129; p=0.043). However, no statistically significant dependence on the prevention of COVID-19 incidence based on the type of vaccines used was found (Kruskal-Wallis χ2=2.072; p=0.72). A statistically significant difference in C-reactive protein levels is observed between groups with early mild complications and early moderate-severity complications (DSCF: W=4.193, p=0.009). A statistically significant difference in LDH levels is noted between individuals without chronic diseases and those with chronic diseases at the time of the study (Kruskal-Wallis χ2=6.08, p=0.014). In individuals vaccinated against the SARS-CoV-2 virus, a positive correlation is found between the levels of C-reactive protein and lactate dehydrogenase (Kendall's τb 0.134, p < 0.001). The mean levels of lactate among individuals with mild, moderate, and severe forms of COVID-19 are higher than the reference mean; similarly, the mean levels of glucose in these same groups are higher than the reference mean. A positive correlation exists between the levels of lactate and glucose among individuals vaccinated against the SARS-CoV-2 virus (Kendall's τb 0.082, p < 0.01). Conclusions. Vaccination contributes to an increase in antibody levels. The level of antibodies after the third vaccination exceeded the levels after the first (Dwass-Steel-Critchlow-Fligner (DSCF): W 4.42, p=0.005) and second vaccinations (W 4.24, p=0.008). Vaccination reduces the likelihood of COVID-19 infection by 1.84 times (Odds Ratio ‒ 1.84; 95% Confidence Interval 1.02‒3.30; Pearson χ2=4.129; p=0.043). The frequency of COVID-19 incidence is not dependent on the type of vaccine used: AstraZeneca, Comirnaty, CoronaVac, Moderna, Pfizer (Kruskal-Wallis χ2=2.072; p=0.723), and the level of antibodies in the vaccinated individuals' serum. In the post-COVID-19 remote period, regardless of vaccination status, various complications are observed. However, among the vaccinated, the number of individuals without complications or with minimal complications is greater than in the unvaccinated group, while the number of individuals with early and severe complications is lower (Kruskal-Wallis χ2=6.127; p=0.047). A high level of C-reactive protein (DSCF: W=4.19, p=0.009), a tendency toward increased levels of lactate dehydrogenase (DSCF: W=3.27, p=0.054), elevated levels of lactate (2.17+1.23, t=3.34; p=0.002), and glucose (6.06+0.048, t=10.54; p < 0.001) indicate that after recovering from COVID-19, regardless the type of vaccines used, in individuals with distant symptoms there are metabolic changes that are signs of a chronic inflammatory process. Individuals with chronic diseasees show an increase in the level of lactate dehydrogenase (χ2=6.08; p=0.014) and a tendency toward increased levels of C-reactive protein (χ2=3.74; p=0.053). Molecular markers of inflammation such as increased levels of lactate, glucose, C-reactive protein, and lactate dehydrogenase are informative for identifying individuals with an inflammatory process in the post-COVID-19 remote period.

Decades Long Involvement of THP-1 Cells as a Model for Macrophage Research: A Comprehensive Review.

Over the years, researchers have endeavored to identify dependable and reproducible in vitro models for examining macrophage behavior under controlled conditions. The THP-1 cell line has become a significant and widely employed tool in macrophage research within these models. Originating from the peripheral blood of individuals with acute monocytic leuke-mia, this human monocytic cell line can undergo transformation into macrophage-like cells, closely mirroring primary human macrophages when exposed to stimulants. Macrophages play a vital role in the innate immune system, actively regulating inflammation, responding to infec-tions, and maintaining tissue homeostasis. A comprehensive understanding of macrophage bi-ology and function is crucial for gaining insights into immunological responses, tissue healing, and the pathogenesis of diseases such as viral infections, autoimmune disorders, and neoplastic conditions. This review aims to thoroughly evaluate and emphasize the extensive history of THP-1 cells as a model for macrophage research. Additionally, it will delve into the significance of THP-1 cells in advancing our comprehension of macrophage biology and their invaluable contributions to diverse scientific domains.

CD4+ T Cells From Individuals With Type 1 Diabetes Respond to a Novel Class of Deamidated Peptides Formed in Pancreatic Islets.

The β-cell plays a crucial role in the pathogenesis of type 1 diabetes, in part through the posttranslational modification of self-proteins by biochemical processes such as deamidation. These neoantigens are potential triggers for breaking immune tolerance. We report the detection by LC-MS/MS of 16 novel Gln and 27 novel Asn deamidations in 14 disease-related proteins within inflammatory cytokine-stressed human islets of Langerhans. T-cell clones responsive against one Gln and three Asn deamidated peptides could be isolated from peripheral blood of individuals with type 1 diabetes. Ex vivo HLA class II tetramer staining detected higher T-cell frequencies in individuals with the disease compared to control individuals. Furthermore, there was a positive correlation between the frequencies of T cells specific for deamidated peptides, insulin antibody levels at diagnosis, and duration of disease. These results highlight that stressed human islets are prone to enzymatic and biochemical deamidation and suggest that both Gln and Asn deamidated peptides can promote the activation and expansion of autoreactive CD4+ T cells. These findings add to the growing evidence that post-translational modifications undermine tolerance and may open the road for the development of new diagnostic and therapeutic applications for individuals living with type 1 diabetes.

Targeted Genomic Sequencing of TSC1 and TSC2 Reveals Causal Variants in Individuals for Whom Previous Genetic Testing for Tuberous Sclerosis Complex Was Normal

Tuberous sclerosis complex (TSC) is caused by inactivating variants in TSC1 and TSC2. Somatic mosaicism, as well as the size and complexity of the TSC1 and TSC2 loci, makes variant identification challenging. Indeed, in some individuals with a clinical diagnosis of TSC, diagnostic testing fails to identify an inactivating variant. To improve TSC1 and TSC2 variant detection, we screened the TSC1 and TSC2 genomic regions using targeted HaloPlex custom capture and next-generation sequencing (NGS) in genomic DNA isolated from peripheral blood of individuals with definite, possible or suspected TSC in whom no disease-associated variant had been identified by previous diagnostic genetic testing. We obtained >95% target region coverage at a read depth of 20 and >50% coverage at a read depth of 300 and identified inactivating TSC1 or TSC2 variants in 83/155 individuals (54%); 65/113 (58%) with clinically definite TSC and 18/42 (43%) with possible or suspected TSC. These included 19 individuals with deep intronic variants and 54 likely cases of mosaicism (variant allele frequency 1-28%; median 7%). In 13 cases (8%), we identified a variant of uncertain significance (VUS). Targeted genomic NGS of TSC1 and TSC2 increases the yield of inactivating variants found in individuals with suspected TSC.

The Use of Novel Antibodies to Characterize B cells in Health and Disease

IntroductionCD38, a glycoprotein ectoenzyme found on the surface of majority of hematopoietic cells, can exist in monomeric or multimeric conformations and enzymatic activity correlates with multimerization. During human B cell differentiation, CD38 expression levels are low on naïve and memory B cells, intermediate on transitional cells, and high on antibody-secreting plasmablasts and plasma cells. Conventional anti-CD38 (C-CD38) antibodies, recognize all CD38 formations and are not selective for identification of plasmablasts, thus requiring the use of additional antibodies including CD27. In contrast, a novel sea lamprey derived monoclonal VLRB antibody, tetVLRB MM3 (V-CD38), recognizes an epitope likely located within the catalytic site of active multimeric CD38 and therefore might be more specific to plasmablasts, which express active CD38. ObjectivesCharacterize the B cells recognized by V-CD38 in the peripheral blood of individuals with or without inborn errors of immunity (IEI). MethodsB cell subsets were characterized as described in figure 1 using conventional antibodies to CD3, CD19, IgM, CD27, C-CD38, and by V-CD38 generated by a collaborator. Results were presented as means with standard deviations (SD). Correlations were calculated using the Pearson’s Correlation Coefficient formula. ResultsTo validate our identification of plasmablasts with V-CD38, samples from patients with X-linked agammaglobulinemia lacking B cells, and tonsils rich in plasmablasts were used as negative and positive controls, respectively. Analysis of samples from 16 individuals revealed nearly identical (r = 0.99, p <.00001) proportions of CD27hi plasmablasts recognized by V-CD38 (0.51%+/-0.74%) and C-CD38 (0.49%+/-0.70%), suggesting that all cells expressed multimeric CD38. In contrast, only 45.28% ±16.19% of transitional IgMhighCD27-C-CD38int B cells were also recognized by V-CD38, (2.52%+/–3.62% and 1.15%+/–1.75%, respectively), a finding that was consistent across samples (r = 0.98, p <.00001). Preliminary studies in three IEI patients demonstrated markedly reduced plasmablasts (0.035%+/–0.026%), class-switched memory B cells (1.25% +/–1.52%) and transitional cells (1.25%+/–1.06%) compared to healthy controls (0.59%+/–0.74%, 15.29%+/–8.95%, and 2.81%+/–3.96% respectively). [Display omitted] ConclusionsWhile C-CD38 and V-CD38 antibodies can identify plasmablasts in peripheral blood, V-CD38 allows more selective detection of CD38 on plasmablasts. Some circulating transitional cells also express multimeric CD38. Additional studies will determine whether V-CD38 can better identify patients suffering from abnormal plasmablast formation and impaired production of antibodies.

Investigation of liquid biopsy analytes in peripheral blood of individuals after SARS-CoV-2 infection

Post-acute COVID-19 syndrome (PACS) is linked to severe organ damage. The identification and stratification of at-risk SARS-CoV-2 infected individuals is vital to providing appropriate care. This exploratory study looks for a potential liquid biopsy signal for PACS using both manual and machine learning approaches. Using a high definition single cell assay (HDSCA) workflow for liquid biopsy, we analysed 100 Post-COVID patients and 19 pre-pandemic normal donor (ND) controls. Within our patient cohort, 73 had received at least 1 dose of vaccination prior to SARS-CoV-2 infection. We stratified the COVID patients into 25 asymptomatic, 22 symptomatic COVID-19 but not suspected for PACS and 53 PACS suspected. All COVID-19 patients investigated in this study were diagnosed between April 2020 and January 2022 with a median 243 days (range 16-669) from diagnosis to their blood draw. We did a histopathological examination of rare events in the peripheral blood and used a machine learning model to evaluate predictors of PACS. The manual classification found rare cellular and acellular events consistent with features of endothelial cells and platelet structures in the PACS-suspected cohort. The three categories encompassing the hypothesised events were observed at a significantly higher incidence in the PACS-suspected cohort compared to the ND (p-value < 0.05). The machine learning classifier performed well when separating the NDs from Post-COVID with an accuracy of 90.1%, but poorly when separating the patients suspected and not suspected of PACS with an accuracy of 58.7%. Both the manual and the machine learning model found differences in the Post-COVID cohort and the NDs, suggesting the existence of a liquid biopsy signal after active SARS-CoV-2 infection. More research is needed to stratify PACS and its subsyndromes. This work was funded in whole or in part by Fulgent Genetics, Kathy and Richard Leventhal and Vassiliadis Research Fund. This work was also supported by the National Cancer InstituteU54CA260591.

Estimation of hematological and biochemical parameters of a representative of the amphibious fauna of the Urals: Salamandrella keyserlingii (Caudata, Amphibia)

The paper has presented for the first time the results of a study of hematological and biochemical parameters of a threatened species of the fauna of the Middle Urals, Salamandrella keyserlingii (Caudata, Amphibia). Mature males of S. keyserlingii (n = 30; sampled in spring and summer) living in the Protected Area «Kalinovsky Forest Park» (Sverdlovsk Region, Russia), located in the urban agglomeration of Yekaterinburg city, were used as research objects. The leukocyte composition of S. keyserlingii blood is represented by granulocytes (neutrophils: early, bands, segments; basophils, eosinophils) and agranulocytes (monocytes and lymphocytes). The peripheral blood of individuals was characterised by the predominance of agranulocytes. They provide immune surveillance and selective reactivity of the organism (adaptive immunity) both in spring (55.7%) and summer (60.3%) seasons (p &lt; 0.001). The content of granulocytes, providing non-specific urgent protection of the organism from infections and toxic effects, is ranged as 39.3–44.4% in males. In the spring and summer periods, the leukocyte formula of the peripheral blood of S. keyserlingii has a high lymphoid profile (52.4–56.7%). Seasonal variability of the integral leukocyte index (ISL) of S. keyserlingii was 0.80 in spring and 0.65 in summer. The content of free amino acids in the blood plasma of S. keyserlingii has been assessed for the first time from the standpoint of their participation in the processes of immunomodulation and the formation of adaptive reactions that ensures the survival and eurybiont of the species in a wide temperature range. The amino acid fund of S. keyserlingii blood plasma was represented by 17 amino acids: alanine, serine, threonine, lysine, leucine, valine, glycine, aspartic acid, glutamic acid, arginine, histidine, methionine, proline, phenylalanine, cystine, tyrosine, and isoleucine. The analysis of the metabolic groups of amino acids in the blood plasma of S. keyserlingii shows a high percentage of glycogenic amino acid (GGAA: 82.1%) and essential amino acids (EAA: 53.4%). The high content of amino acids in the blood indicates their relevance in the processes of protein synthesis, active participation in both immune processes and in the processes of detoxification and the formation of adaptive reactions during the summer period of amphibian’s growth and development. The obtained results of studies of hematological and biochemical parameters of S. keyserlingii’s blood significantly expand the understanding of the adaptation mechanisms of amphibians to conditions of natural and anthropogenic habitat transformations. This is undoubtedly of theoretical interest and applies significance for development of environmental measures in the monitoring system of natural and artificial aquatic ecosystems.

Залежність рівня ендотеліну-1 у осіб, що перенесли негоспітальну пневмонію на тлі коронавірусної хвороби (COVID-19), від особливостей перебігу гострого й постгострого періодів

Abstract. The impairment of endothelial function is one of the mechanisms contributing to the increased risk of cardiovascular and cerebrovascular events in patients who have experienced community-acquired pneumonia associated with COVID-19. The aim of the study was to establish the connection between the level of endothelin-1 in peripheral blood of individuals who have experienced community-acquired pneumonia associated with COVID-19, with clinical-anamnestic, demographic, laboratory, and functional indicators in both the acute and post-acute periods. Materials and methods. 39 patients (age — 57.0 (48.5; 64.0) years, male — 18 (46.2 %), female — 21 (53.8 %)) survived COVID-19-associated community-acquired pneumonia were investigated. They were investigated on the 45.0 (40.0; 60.0) day from the onset of COVID-19 and had the dyspnea at the moment of the investigation. The examination included analyzing complaints, medical history and documentation, physical examination, pulse oximetry (SpO2 ), a 6-minute walk test (6MWT), measurement of lung diffusion capacity (DLco), laboratory methods (C-reactive protein (CRP), D-dimer, platelet count, endothelin-1 measurements). Results. Since it was found that patients, on one hand, had varying degrees of respiratory failure, different laboratory parameters during the acute period of COVID-19, and varying severity of dyspnea in the post-acute period of the disease, and on the other hand, exhibited wide fluctuations in endothelin-1 levels, a cluster analysis was conducted. Two clusters of patients were identified. Cluster 1 comprised 15 individuals with higher body mass index, lower SpO2 levels at the time of examination, and poorer results in the 6MWT and DLco compared to Cluster 2, which included 24 individuals. The level of endothelin-1 in Cluster 1 was significantly higher than that in Cluster 2. Clusters 1 and 2 were comparable in terms of age and gender but differed in terms of treatment conditions (outpatient or inpatient), duration of hospitalization, minimum SpO2 levels, CRP, and D-dimer levels in the acute period, and severity of the acute phase of COVID-19. Conclusion. A high level of endothelin-1 in patients who have experienced severe or critical COVID-19, accompanied by elevated levels of CRP and D-dimer, overweight or obesity, significant dyspnea, low tolerance to physical exertion, and poor DLco in post-acute period of COVID-19, indicates the necessity of careful monitoring of these individuals to prevent the development of adverse events in post-acute period of COVID-19.

Utility of Peripheral Blood Flow Cytometric Blast Enumeration Reporting in Patients without Prior Hematologic Malignancy

Background: The significance of documenting the enumerated data of low levels of blast events (CD34+/CD117+) by peripheral blood flow cytometry studies is largely unaddressed in the literature. The presence of a blast population in peripheral blood of individuals without documented appropriate explanation (such as treatment with a colony-stimulating factor) can be the very first indication of an evolving hematolymphoid neoplastic process. We aim to identify a threshold of quantified blast events on peripheral blood flow cytometric studies that may alert medical professionals of a possible disease process affecting the bone marrow. Methods and materials: Peripheral blood flow cytometry studies performed by the clinical flow cytometry laboratory at the University of California Davis Medical Center between 2015 and 2020 that were detected to have less than or equal to 1% blast population were evaluated. Patients with a history of a known hematologic malignancy at the time of the aforementioned peripheral blood flow cytometry study were excluded. The remaining cases from patients without a known hematological malignancy were investigated. All flow cytometry events were analyzed by multiparametric 10-color flow cytometry using an extensive panel of monoclonal antibodies. The data events were analyzed with Kaluza® software. Percentages of blasts in patients with progression to a hematologic malignancy were compared with the group of patients without progression to a hematologic malignancy. The results of this comparison were utilized to establish a threshold of the presence of CD34+/CD117+ blasts that may warrant formal reporting and recommendations of additional studies. Results: A total of 165 flow cytometry cases from 2015-2020 were evaluated. The patients' electronic medical records were reviewed in order to determine the presence of a prior hematolymphoid malignancy and to evaluate for the progression of a malignancy after the flow study. Of the cases reviewed, 110 were excluded due to the presence of a prior hematolymphoid malignancy. The remaining 55 cases were separated into a group who on follow up had no progression to a hematolymphoid malignancy (n=50), and a second group who developed a hematolymphoid malignancy (n=5) (see figure). The mean percent of CD34+/CD117+ events in the group of patients who progressed to a hematolymphoid malignancy was higher than the non-malignancy control group (0.80% vs 0.40% of total flow cytometry events respectively). The mean percent of CD34+ events in the group of patients who had a malignancy was slightly higher than the control non-malignant group mean percent CD34+ events (0.248% vs 0.220% of total flow cytometry events respectively). Conclusions: We identified a small percentage of patients with no known hematolymphoid malignancy and low levels of blasts on peripheral blood flow cytometry study who later developed a hematolymphoid malignancy (5/55 = 9%). On further investigation, a slightly elevated level of CD34+/CD117+ blast events in these patients was noted. Of the patients who did not develop hematolymphoid malignancies, a significant percentage had non-malignant inflammatory conditions. Based on our current assessment, some patients with low levels of circulating blasts detected by flow cytometry may necessitate follow-ups, including bone marrow evaluation. The presence of cytopenias and/or elevated inflammatory markers may further assist in the decision to proceed with additional hematologic work-up. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Clonal IgA and IgG autoantibodies from individuals at risk for rheumatoid arthritis identify an arthritogenic strain of Subdoligranulum.

The mucosal origins hypothesis of rheumatoid arthritis (RA) proposes a central role for mucosal immune responses in the initiation or perpetuation of the systemic autoimmunity that occurs with disease. However, the connection between the mucosa and systemic autoimmunity in RA remains unclear. Using dual immunoglobulin A (IgA) and IgG family plasmablast-derived monoclonal autoantibodies obtained from peripheral blood of individuals at risk for RA, we identified cross-reactivity between RA-relevant autoantigens and bacterial taxa in the closely related families Lachnospiraceae and Ruminococcaceae. After generating bacterial isolates within the Lachnospiraceae/Ruminococcaceae genus Subdoligranulum from the feces of an individual, we confirmed monoclonal antibody binding and CD4+ T cell activation in individuals with RA compared to control individuals. In addition, when Subdoligranulum isolate 7 but not isolate 1 colonized germ-free mice, it stimulated TH17 cell expansion, serum RA-relevant IgG autoantibodies, and joint swelling reminiscent of early RA, with histopathology characterized by antibody deposition and complement activation. Systemic immune responses were likely due to mucosal invasion along with the generation of colon-isolated lymphoid follicles driving increased fecal and serum IgA by isolate 7, because B and CD4+ T cell depletion not only halted intestinal immune responses but also eliminated detectable clinical disease. In aggregate, these findings demonstrate a mechanism of RA pathogenesis through which a specific intestinal strain of bacteria can drive systemic autoantibody generation and joint-centered antibody deposition and immune activation.

Importance of clonal hematopoiesis for hematologic neoplasms

Clonal hematopoiesis of indeterminate potential (CHIP) is afairly newly described phenomenon characterized by myeloid cancer-associated somatic mutations detectable in the peripheral blood of individuals without evidence of hematologic disease. Individuals with CHIP have asignificantly increased risk of developing ahematologic malignancy, although the overall rate of transformation is low. We review the current state of knowledge on causes of clonal expansion of blood cells as well as identifiable risk factors for progression to overt hematologic malignancy. CHIP is considered apremalignant state and predisposes to the development of hematologic malignancy. Because the overall rate of transformation is low, clear identification and subsequent monitoring of those CHIP individuals at a higher risk is of paramount importance. In the future, prospective studies evaluating preventive and/or preemptive therapeutic strategies may aid in avoiding progression to blood cancer in individuals with CHIP.

High baseline expression of IL-6 and IL-10 decreased CCR7 B cells in individuals with previous SARS-CoV-2 infection during BNT162b2 vaccination.

The current pandemic generated by SARS-CoV-2 has led to mass vaccination with different biologics that have shown wide variations among human populations according to the origin and formulation of the vaccine. Studies evaluating the response in individuals with a natural infection before vaccination have been limited to antibody titer analysis and evaluating a few humoral and cellular response markers, showing a more rapid and intense humoral response than individuals without prior infection. However, the basis of these differences has not been explored in depth. In the present work, we analyzed a group of pro and anti-inflammatory cytokines, antibody titers, and cell populations in peripheral blood of individuals with previous SARS-CoV-2 infection using BNT162b2 biologic. Our results suggest that higher antibody concentration in individuals with an earlier disease could be generated by higher production of plasma cells to the detriment of the presence of memory B cells in the bloodstream, which could be related to the high baseline expression of cytokines (IL-6 and IL-10) before vaccination.

Charting the proteome landscape in major psychiatric disorders: From biomarkers to biological pathways towards drug discovery

Schizophrenia (SZ), bipolar disorder (BD), and major depressive disorder (MDD) are major mental disorders that affect a significant proportion of the global population. Advancing our knowledge of the pathophysiology of these disorders and identifying biomarkers are urgent needs for developing objective diagnostic tests and new therapeutics. In this study, we performed a systematic review and then extracted, curated, and analyzed proteomics data from published studies, aiming to assess the proteome in peripheral blood of individuals with SZ, BD, or MDD. Then, we performed pathway and network analyses to illuminate the biological themes concatenated by the differentially expressed proteins by systematically interrogating the literature to uncover biological pathways with more robust biological meaning. We identified 486 differentially expressed proteins from 51 studies across the three disorders with 9,423 participants. The great majority of pathways were common to SZ, BD, and MDD. They were related to the immune system, including signaling by interleukins, Toll-like receptor signaling pathway, and complement cascade, and to signal transduction, notably MAPK1/MAPK3 signaling, PI3K-Akt Signaling Pathway, Focal Adhesion-PI3K-Akt-mTOR-signaling, rhodopsin-like receptors, GPCR signaling, and the JAK-STAT signaling pathway. Other shared pathways included advanced glycosylation end-product receptor signaling, Regulation of Insulin-like Growth Factor, cholesterol metabolism, and IL-17 signaling pathway. Pathways shared between SZ and BD were integrin cell-surface interactions, GRB2:SOS provides linkage to MAPK signaling for integrins, and syndecan interactions. Shared between BD and MDD were the NRF2 pathway and signaling by EGFR pathways. Our findings advance our understanding of the protein variations and associations with these disorders, which are useful for accelerating biomarker development and drug discovery.

Субпопуляционный состав Т-хелперов в периферической крови хронически облученных лиц в отдаленном периоде

Earlier, it has been convincingly established that exposure to ionizing radiation (IR) alters the T cell-mediated immunity in the long term. However, a search for papers describing the effect chronic exposure to radiation has on various subpopulations of T-helpers yielded no results. Therefore, we designed this study seeking to investigate the quantitative characteristics of various subpopulations of T-helpers in the peripheral blood of individuals chronically exposed to low-level radiation for a long period of time. The study involved 102 chronically exposed Techa Riverside residents (Russia) aged 60–87 years. The participants were divided into two groups, one comprised of exposed individuals with the average red bone marrow (RBM) irradiation dose of 567 ± 73 mGy, another, the control group, comprised of people with the irradiation dose below 70 mGy. With the help of flow cytometry, we identified the quantitative characteristics of T-helper subpopulations in the peripheral blood at various stages of their differentiation, as well as various T-helper subpopulations of central and effector memory. The study revealed no significant differences in the composition of T-helper subpopulations in the compared groups. We discovered a significant growth of the double positive follicular T-helper 17 subpopulation in the population of central memory T-helpers, which is associated with the increase of RBM (p = 0.04; S = 0.19), thymus and peripheral lymphoid organs (p = 0.03; S = 0.22) irradiation dose. In the group of exposed individuals, the number of naive T-helpers (p = 0.009) and double positive follicular T-helpers 17 in the TEM subpopulation (p = 0.04) was decreasing as the age of participants increased, and the number of effector memory T-helpers, on the contrary, increased with age (p = 0.04). We have not registered similar phenomena in the comparison group.

The Differential Expression of Circular RNAs in Type 2 Diabetes Mellitus and Latent Autoimmune Diabetes in Adults.

Objective: Expression of circular RNAs (circRNAs) in the peripheral blood of individuals with latent autoimmune diabetes in adults (LADA) and type 2 diabetes mellitus (T2DM) were quantified to identify dysregulated circRNAs compared with control individuals. Methods: circRNAs were obtained from the peripheral blood serum of 12 healthy adults and 12 individuals with LADA and 12 type 2 diabetics. The circRNA expression profiles were analyzed by high-throughput RNA sequencing. The most highly dysregulated circular RNAs were validated by quantitative real-time polymerase chain reaction. A circular RNA-microRNA (miRNA) network diagram predicted the interactions of circular RNAs, miRNAs, and coding genes. Results: A total of 2334 differentially expressed circRNAs were detected among the three groups, with 277 circRNAs in the Group DM versus Group NG; 992 circRNAs in the Group LADA versus Group NG and 1065 circRNAs in the Group DM versus Group LADA. Six circRNAs were identified as the most distinctive differentially expressed targets (p < 0.05). The proposed molecular functions of these differentially expressed circRNAS included the tumor necrosis factor signaling pathway, the FoxO signaling pathway, cellular senescence, and long-term potentiation (all false discovery rate p < 0.05) which may contribute to T2DM and LADA. Conclusion: circRNAs are aberrantly expressed in the peripheral blood of patients with T2DM and LADA and may interact with miRNA and circRNA-derived peptides in the development of diabetes. Further investigations may illustrate the partial pathogenesis of diabetes mellitus. Clinical Trial Registration number: ChiCTR1900020644.

Transcript levels of cytokine coding genes in peripheral blood and tissues of patients with periodontitis.

Periodontal diseases are common conditions in almost all age groups and a public health problem. Numerous risk factors have been demonstrated for this condition. The main mechanism of tissue destruction in the periodontitis is the functional interactions between microbial pathogens and host immune responses, thus cytokines have crucial roles in the pathogenesis periodontitis. Our previous study has demonstrated the susceptibility role of HLA-DRB1*04 allele in development of this disease. So, the individuals who were positive for HLA-DRB1*04 allele were excluded. We aimed to appraise the function of cytokines in the pathogenesis of periodontitis via assessment of tissue and blood levels of a number of cytokine coding genes, namely IL-1B, CXCL8, IL-17, IFNG, TGFB and TNFA1. Expressions of IFNG, IL-17, TGFB and TNFA1 were significantly higher in the peripheral blood of individuals with periodontitis compared with unaffected persons (Posterior beta = 1.91, P value = 0.043; Posterior beta = 1.84, P value = 0.033; Posterior beta = 0.713, P value = 0.009 and Posterior beta = 2.85, P value = 0.001, respectively). Moreover, expression of IL-17 was higher in females compared with males (Posterior beta = 1.47, P value = 0.036). As the interaction effect between gender and group was remarkable for IL-17 expression, we further conducted subgroup analysis within gender group. Expression of IL-17 was higher in male patients compared with unaffected males (Posterior beta = 1.9, P value = 0.048). We did not detect any significant difference in the expression of these cytokines in tissues obtained from affected individuals and unaffected controls. Therefore, our results imply dysregulation of cytokine coding genes in patients with periodontitis and warrant further mechanistical studies.

Gut microbiota\u2013derived metabolite 3-idoleacetic acid together with LPS induces IL-35+ B cell generation

BackgroundIL-35–producing Bregs and Treg cells critically regulate chronic illnesses worldwide via mechanisms related to disrupting the gut microbiota composition. However, whether the gut microbiota regulates these IL-35+ cells remains elusive. We herein investigated the regulatory effects of the gut microbiota on IL-35+ cells by using genetically modified mouse models of obesity.ResultsWe first found that gut Reg4 promoted resistance to high-fat diet-induced obesity. Using 16S rRNA sequencing combined with LC-MS (liquid chromatography–mass spectrometry)/MS, we demonstrated that gut Reg4 associated with bacteria such as Lactobacillus promoted the generation of IL-35+ B cells through 3-idoleacetic acid (IAA) in the presence of LPS. HuREG4IECtg mice fed a high-fat diet exhibited marked IL-35+ cell accumulation in not only their adipose tissues but also their colons, whereas decreased IL-35+ cell accumulation was observed in the adipose and colon tissues of Reg4 knockout (KO) mice. We also found that Reg4 mediated HFD-induced obesity resistance via IL-35. Lower levels of IAA were also detected in the peripheral blood of individuals with obesity compared with nonobese subjects. Mechanistically, IAA together with LPS mediated IL-35+ B cells through PXR and TLR4. KO of PXR or TLR4 impaired the generation of IL-35+ B cells.ConclusionTogether, IAA and LPS induce the generation of IL-35+ B cells through PXR and TLR4.ESFDmA-1LKhrrc6QfnM_pPVideo

Presence of periodontal pathogenic bacteria in blood of patients with coronary artery disease

It has been hypothesised that oral bacteria can migrate, through the blood, from the mouth to the arterial plaques, thus exacerbating atherosclerosis. This study compared bacteria present in the peripheral blood of individuals with and without coronary artery disease (CAD). RNA sequences obtained from blood were downloaded from GEO (GSE58150). Eight patients with coronary artery calcification (CAC) scoring > 500 and eight healthy individuals were analysed. After conducting quality control, the sequences were aligned to the hg38 reference genome using Hisat2. Bacterial taxa were analysed by inputting the unmapped sequences into Kraken. Ecological indices were calculated using Vegan. The package DESeq2 was used to compare the counts of bacteria per standard rank between groups. A total of 51 species were found only in patients with CAD and 41 were exclusively present in healthy individuals. The counts of one phylum, one class, three orders, two families and one genus were significantly different between the analysed groups (p < 0.00032, FDR < 10%), including the orders Cardiobacteriales, Corynebacteriales and Fusobacteriales. Twenty-three bacterial species belonging to the subgingival plaque bacterial complexes were also identified in the blood of individuals from both the groups; Fusobacterium nucleatum was significantly less frequent in patients with CAD (p = 0.0012, FDR = 4.8%). Furthermore, the frequency of another 11 bacteria differed significantly among patients with CAD than that among healthy individuals (p < 0.0030, FDR < 10%). These bacteria have not been previously reported in patients with atherosclerosis and periodontitis. The presence of members of the subgingival plaque bacterial complexes in the blood of patients with CAC supports the hypothesis that the periodontopathogens can be disseminated through the blood flow to other body parts where they may enhance inflammatory processes that can lead to the development or exacerbation of atherosclerosis.

Genetic polymorphism of opioid receptors and use of opioid substances by high school students of Cotonou and Parakou (Benin)

In Benin, schools become sometimes the hub for dealing and consuming psychoactive substances; opioids in particular. The objectives of this study are to identify the risk factors related to the use of opioids and investigate the genetic polymorphism of mu and delta opioid receptors of teenagers and young adults who consume opioids in schools. To accomplish this, 453 students participated in this study; R diversity 3.6.1 software in the RStudio environment was used to identify students who experience opioids through ASSIST V3.0 scoring. SNPs A118G on the OPRM gene (µ) and T921C on the OPRD gene (d), were searched by PCR on DNA extracts from peripheral blood of individuals. We identified 54 regular opioid users and 399 non users. This experience begins for most with the consumption of alcohol and tobacco and is facilitated by the proximity of marshlands, kiosks, and pubs near high schools and colleges. The aggressive advertisement combined with relative socio-cultural tolerance just worsen this behavioral deviance. We found no difference in the SNP frequencies of the OPRM (µ) and OPRD (d) genes between students opioids consumers and non-consumers.

Vascular repair and regeneration in cardiometabolic diseases.

Chronic cardiometabolic assaults during type 2 diabetes (T2D) and obesity induce a progenitor cell imbalance in the circulation characterized by overproduction and release of pro-inflammatory monocytes and granulocytes from the bone marrow alongside aberrant differentiation and mobilization of pro-vascular progenitor cells that generate downstream progeny for the coordination of blood vessel repair. This imbalance can be detected in the peripheral blood of individuals with established T2D and severe obesity using multiparametric flow cytometry analyses to discern pro-inflammatory vs. pro-angiogenic progenitor cell subsets identified by high aldehyde dehydrogenase activity, a conserved progenitor cell protective function, combined with lineage-restricted cell surface marker analyses. Recent evidence suggests that progenitor cell imbalance can be reversed by treatment with pharmacological agents or surgical interventions that reduce hyperglycaemia or excess adiposity. In this state-of-the-art review, we present current strategies to assess the progression of pro-vascular regenerative cell depletion in peripheral blood samples of individuals with T2D and obesity and we summarize novel clinical data that intervention using sodium-glucose co-transporter 2 inhibition or gastric bypass surgery can efficiently restore cell-mediated vascular repair mechanisms associated with profound cardiovascular benefits in recent outcome trials. Collectively, this thesis generates a compelling argument for early intervention using current pharmacological agents to prevent or restore imbalanced circulating progenitor content and maintain vascular regenerative cell trafficking to sites of ischaemic damage. This conceptual advancement may lead to the design of novel therapeutic approaches to prevent or reverse the devastating cardiovascular comorbidities currently associated with T2D and obesity.