Immunological Reactions Research Articles

Key platelet genes play important roles in predicting the prognosis of sepsis

Sepsis is a life-threatening organ malfunction induced by an imbalanced immunological reaction to infection in the host. Many studies have utilized traditional RNA sequencing (RNA-seq) data to identify important biological targets to predict sepsis prognosis. However, alterations in core cells and functional status cannot be effectively detected in sepsis patients. The goal of this study was to identify key cells through single-cell RNA-seq (scRNA-seq), and combine bulk RNA-seq data and multiple algorithm analysis to construct a stable prognostic model for sepsis. The scRNA-seq and bulk RNA-seq data from sepsis patients were collected from the Gene Expression Omnibus (GEO) database. The R package “Seurat” was used to process the scRNA-seq data. Cell communication was investigated using the R package “CellChat”. The pseudo-time of the cells was calculated using the R package “monocle”. The R package “limma” was used to identify differentially expressed genes (DEGs) between the sepsis group and the control group. Weighted gene correlation network analysis (WGCNA) was used to identify critical modules. Eight kinds of machine learning and 90 algorithm combinations were used to construct the prognostic model for sepsis. Quantitative real-time PCR (qRT‒PCR) was performed to determine the expression of key genes in the cecal ligation and puncture (CLP)-induced sepsis mouse model. The immunological status and related properties of DEGs were then investigated in the high- and low-risk groups delineated by the model. By combining the scRNA-seq data from nine samples, 13 clusters and 9 cell types were identified. CellChat analysis revealed that the number and strength of interactions between platelets and a variety of cells increased. We identified key platelet genes from the scRNA-seq data and combined these genes and the results of differential analysis and WGCNA of the bulk RNA-seq data. After univariate Cox regression analysis, we calculated the Cindex of the model constructed by the combination of 90 algorithms, and we finally determined the “CoxBoost + Lasso” combination. Multivariate Cox regression was used to construct the final prognostic model. The qRT-PCR results revealed significant differences in five key prognostic genes between the CLP and sham groups. The data was classified into high- and low-risk groups based on the model score. The high-risk group had a poorer survival rate and less immune infiltration. We identified the importance of platelets in sepsis patients through scRNA-seq, and established prognostic models with key genes that were identified via scRNA-seq combined with bulk RNA-seq analysis. The results of this model were closely associated with patient survival rates and immunological status and this model is useful for the prognostic management of sepsis.

Clinical Evaluation of Some Biochemical Parameters from Patients in Heamodialysis Room

As a marker of systemic inflammation, raised (C-reactive protein (CRP)) concentrations which are still within the normal range have been associated with an increased inflammation of chronic renal diseases (CRD). The current study aimed to establish potential determinats of raised CRP concentrations in patients who treated in Heamodialysis room,then study the relationship between CRP& some biochemical parameters related CRD We used a CRP latex reagents Kit which is based on an immunological reaction between CRP antisera bounded to the biologically inert latex particles or with CRP in the test specimens of 19 patients with (CRD) mean age 48 years ,range = 30?65 & in 21 healthy subjects as control group their age range = 30 ?45 years. The results are classified according to visible agglutination to: 1-A positive result / is indicated by the obvious agglutination pattern of the latex, in a clear solution. 2-A negative result / is indicated by no change in the latex suspension on the test slide. Then, we correlated the results of the precipitin test with the quantitative data on C-reactive protein. This study has found that: CRP concentrations in patients with CRD were increased very clearly than normal subjects , and established that CRP concentration in male was more than in female for patients specimens. Biochemical studies have shown raising (CRP) concentration is a marker of systemic inflammation. The relation between CRP & S.creatinine ,hemoglobine ,blood sugar is negative ,while positive with blood urea . Other biochemical parameter related to CRD'' blood urea, blood creatinine & hemoglobine and blood sugar'' were assayed for both subjects in order to asses the disease by compared the results. Abbreviation: CRP:C-reactive protein, CRD: Chronic renal diseases, F: Female, M: Male, Hb: hemoglobin, BU: blood Urea, S.Cr: Serum creatinine, BS: blood sugar, nCRP: native CRP, MCRP: Matified CRP.

Enhanced Cancer Biomarker Detection using Fe3O4 Magnetic Nanoparticles: Synthesis, Surface Modifications and Diagnostic Applications: A Review

Fe3O4 magnetic nanoparticles exhibit significant potential for cancer detection due to their unique magnetic properties and versatility. This review examines their role in enhancing cancer biomarker detection, focusing on synthesis methods, surface modifications and diagnostic accuracy. Fe3O4 nanoparticles serve as sensitive MRI contrast agents, facilitating early cancer diagnosis and improving patient prognosis. Synthesis techniques influence their properties, morphology and dimensions, which affect performance. Surface modifications enhance targeting and reduce immunological reactions, enabling precise biomarker detection and effective drug delivery to tumours, minimizing damage to healthy tissue. Despite these advantages, challenges remain in optimizing delivery efficiency and overcoming medication resistance. Further research is required to understand the pharmacokinetics and pharmacodynamics of these nanoparticles, including their distribution, metabolism and physiological impacts. This review provides insights into the potential of Fe3O4 magnetic nanoparticles as cancer detection agents, aiming to guide future research towards developing safer and more efficient applications in medical diagnostics and treatment. HIGHLIGHTS Fe3O4 magnetic nanoparticles exhibit unique magnetic properties that are highly effective in detecting cancer biomarkers. These nanoparticles enable early cancer diagnosis by identifying biomarkers at low concentrations with high sensitivity. The review assesses various synthesis techniques for Fe3O4 magnetic nanoparticles to optimize their detection capabilities. Exploration of surface modification strategies to enhance the functionality and efficacy of Fe3O4 nanoparticles in cancer biomarker detection. Enhanced diagnostic accuracy through the application of Fe3O4 magnetic nanoparticles, promising improved outcomes in cancer detection technologies. GRAPHICAL ABSTRACT

Lateral flow assay sensitivity and signal enhancement via laser µ-machined constrains in nitrocellulose membrane

Lateral flow assay (LFA) is a handful diagnostic technology that can identify severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and other common respiratory viruses in one strip, which can be tested at the point-of-care without the need for equipment or skilled personnel outside the laboratory. Although its simplicity and practicality make it an appealing solution, it remains a grand challenge to substantially enhance the colorimetric LFA sensitivity. In this work, we present a straightforward approach to enhance the sensitivity of LFA by imposing the flow constraints in nitrocellulose (NC) membranes via a number of vertical femtosecond laser micromachined microchannels which is important for prolonged specific binding interactions. Porous NC membrane surfaces were structured with different widths and densities µ-channels employing a second harmonic of the Yb:KGW femtosecond laser and sample XYZ translation over a microscope objective-focused laser beam. The influence of the microchannel parameters on the vertical wicking speed was evaluated from the video recordings. The obtained results indicated that µ-channel length, width, and density in NC membranes controllably increased the immunological reaction time between the analyte and the labeled antibody by 950%. Image analysis of the colorimetric indicators confirmed that the flow rate delaying strategy enhanced the signal sensitives by 40% compared with pristine NC LFA.

Immunological reactivity in cows under technogenic load conditions

Abstract Technogenic load has a negative impact on agricultural efficiency and threatens global food security. Exogenous toxicants have a complex effect on the organism of animals, provoking the development of compensatory mechanisms and changes in the clinical status, morphological and immune blood parameters. The authors analysed the state of the immunohaematological profile. For qualitative and quantitative adaptation reactions characterisation of the organism, leukocyte blood indexes were calculated. The results of the study have allowed to determine the main markers of impaired immunological reactivity in animals kept in territories with unfavourable environmental conditions.

Air pollution exposure and inflammatory bowel disease: a systematic literature review of epidemiological and mechanistic studies

This systematic literature review investigates the relationship between air pollution exposure and inflammatory bowel disease (IBD), encompassing Crohn's disease (CD) and ulcerative colitis (UC). Despite the growing concern over air pollution's impact on various health outcomes, studies specifically addressing effects on the digestive system, particularly IBD, are relatively rare. This review aims to synthesize the current knowledge on this topic, focusing on the mechanisms underlying these associations and the role of different air pollutants. Following PRISMA guidelines, a systematic literature search of PubMed and Web of Science databases was conducted, yielding 13 epidemiological studies and six mechanistic (toxicological) studies meeting the inclusion criteria. The epidemiological studies examined associations between IBD and various air pollutants, including PM2.5 (particles with an aerodynamic diameter smaller than, or equal to, 2.5 µm), PM10 (particles with an aerodynamic diameter smaller than, or equal to, 10 µm), PM2.5–10 (coarse particles with an aerodynamic diameter in the range of 2.5–10 µm), BC (black carbon), NO2 (nitrogen dioxide), NO (nitrogen monoxide), NOx (nitrogen oxides), N2O (nitrous oxide), CO (carbon monoxide), SO2 (sulfur dioxide), VOC (volatile organic compounds), O3 (ozone), Ox (oxidant capacity), and traffic load. Study methodologies varied among these 13 epidemiological studies, including four cohort studies, two ecological studies, three case-control studies, two studies using two-sample Mendelian randomization, and two longitudinal time-series studies. Eight studies investigated associations with Crohn's disease and ulcerative colitis separately, while five studies analyzed IBD as a whole without distinguishing between CD and UC. Eleven studies found statistically significant associations between air pollution exposure and IBD, although inconsistent results were found in several of these studies. A total number of six mechanistic (toxicological) studies were retrieved. Among these six studies, five were using particulate matter as exposure metric, and one was based on NO2 and O3 as exposure metrics. With a combination of animal, human, and in vitro studies, the results in terms of biological mechanisms indicate that air pollution exposure influences the composition of the gut microbiome, altering metabolic functions within the gut, and creates immunological reactions with inflammation contributing to the development of IBD. Consequently, the results suggest a link between air pollution exposure and both the onset and exacerbation of IBD. However, differences in study design, exposure assessment, and pollutant types make it challenging to draw any firm conclusions. Moreover, the lack of multi-pollutant models in most epidemiological studies makes it difficult to estimate the individual effect of specific air pollutants. This review highlights the need for further research utilizing robust study designs and standardized exposure assessment methods to better understand the impact of air pollution on IBD. By elucidating these associations, policymakers and healthcare professionals can develop effective strategies to mitigate the adverse effects of air pollution on digestive health.

A synthetic review: natural history of amniote reproductive modes in light of comparative evolutionary genomics

ABSTRACTThere is a current lack of consensus on whether the ancestral parity mode was oviparity (egg‐laying) or viviparity (live‐birth) in amniotes and particularly in squamates (snakes, lizards, and amphisbaenids). How transitions between parity modes occur at the genomic level has primary importance for how science conceptualises the origin of amniotes, and highly variable parity modes in Squamata. Synthesising literature from medicine, poultry science, reproductive biology, and evolutionary biology, I review the genomics and physiology of five broad processes (here termed the ‘Main Five’) expected to change during transitions between parity modes: eggshell formation, embryonic retention, placentation, calcium transport, and maternal–fetal immune dynamics. Throughout, I offer alternative perspectives and testable hypotheses regarding proximate causes of parity mode evolution in amniotes and squamates. If viviparity did evolve early in the history of lepidosaurs, I offer the nucleation site hypothesis as a proximate explanation. The framework of this hypothesis can be extended to amniotes to infer their ancestral state. I also provide a mechanism and hypothesis on how squamates may transition from viviparity to oviparity and make predictions about the directionality of transitions in three species. After considering evidence for differing perspectives on amniote origins, I offer a framework that unifies (i) the extended embryonic retention model and (ii) the traditional model which describes the amniote egg as an adaptation to the terrestrial environment. Additionally, this review contextualises the origin of amniotes and parity mode evolution within Medawar's paradigm. Medawar posited that pregnancy could be supported by immunosuppression, inertness, evasion, or immunological barriers. I demonstrate that this does not support gestation or gravidity across most amniotes but may be an adequate paradigm to explain how the first amniote tolerated internal fertilization and delayed egg deposition. In this context, the eggshell can be thought of as an immunological barrier. If serving as a barrier underpins the origin of the amniote eggshell, there should be evidence that oviparous gravidity can be met with a lack of immunological responses in utero. Rare examples of two species that differentially express very few genes during gravidity, suggestive of an absent immunological reaction to oviparous gravidity, are two skinks Lampropholis guichenoti and Lerista bougainvillii. These species may serve as good models for the original amniote egg. Overall, this review grounds itself in the historical literature while offering a modern perspective on the origin of amniotes. I encourage the scientific community to utilise this review as a resource in evolutionary and comparative genomics studies, embrace the complexity of the system, and thoughtfully consider the frameworks proposed.

Evaluation of an Enzyme-Linked Magnetic Electrochemical Assay for Hepatitis a Virus Detection in Drinking and Vegetable Processing Water

Globally, waterborne viral infections significantly threaten public health. While current European Union regulations stipulate that drinking water must be devoid of harmful pathogens, they do not specifically address the presence of enteric viruses in water used for irrigation or food production. Traditional virus detection methods rely on molecular biology assays, requiring specialized personnel and laboratory facilities. Here, we describe an electrochemical sandwich enzyme-linked immunomagnetic assay (ELIME) for the detection of the hepatitis A virus (HAV) in water matrices. This method employed screen-printed electrodes as the sensing platform and utilized commercially available pre-activated magnetic beads to provide a robust foundation for the immunological reaction. The ELIME assay demonstrated exceptional analytical performance in only 185 min achieving a detection limit of 0.5 genomic copies per milliliter (g.c./mL) and exhibiting good reproducibility with a relative standard deviation (RSD) of 7% in HAV-spiked drinking and processing water samples. Compared with the real-time RT-qPCR method described in ISO 15216-1, the ELIME assay demonstrated higher sensitivity, although the overall linearity of the method was moderate. These analytical attributes highlight the potential of the ELIME assay as a rapid and viable alternative for HAV detection in water used for agriculture and food processing.

Clinician Perspectives of Gene Therapy as a Treatment Option for Duchenne Muscular Dystrophy.

Duchenne muscular dystrophy (DMD) is a progressive, life-limiting, neuromuscular disorder. Clinicians play an important role in informing families about therapy options, including approved gene therapies and clinical trials of unapproved therapies. This study aimed to understand the perspectives of clinicians about gene therapy for DMD, which has not previously been studied. We conducted interviews with specialist clinicians treating patients with DMD in the United States (n = 8) and United Kingdom (n = 8). Interviews were completed in 2022, before any approved gene therapies, to gain insight into barriers and facilitators to implementing gene therapy and educational needs of clinicians. Most respondents expressed cautious optimism about gene therapy. Responses varied regarding potential benefits with most expecting delayed progression and duration of benefit (1 year to lifelong). Concern about anticipated risks also varied; types of anticipated risks included immunological reactions, liver toxicity, and cardiac or renal dysfunction. Clinicians generally, but not uniformly, understood that gene therapy for DMD would not be curative. Most reported needing demonstrable clinical benefit to justify treatment-related risks. Our data demonstrate variability in knowledge and attitudes about gene therapy among clinicians who follow patients with DMD. As our knowledge base about DMD gene therapy grows, clinician education is vital to ensuring that accurate information is communicated to patients and families.

Assessment of Immunoreactivity Patterns in the Excised Cervical Lymph Nodes of Oral Squamous Cell Carcinoma

Introduction: Epidermoid carcinoma (oral squamous cell carcinoma [OSCC]) comprises approximately 80% of the malignancies of the oral cavity, which is also responsible for the morbidity and mortality due to the distant spread of the neoplastic cells. The regional draining lymph node has been considered an anatomic barrier to the systemic dissemination of malignant cells. It has been postulated that the microscopic architectural pattern of a regional lymph node (RLn) represents the immunologic reactivity in that node and the host response. Morphological assessment of the regional nodes aids in a better understanding of the immune response and its correlation with the stage of the tumour. Materials and Methods: A total of 51 head-and-neck dissection cases were assessed for immunomorphological patterns and the degree of tumour involvement in positive lymph nodes. 49 lymph nodes were in the early stage (TNM Stage l and ll) and 71 lymph nodes were in the advanced stage(TNM stage lll and lV). A total of 595 lymph nodes were histologically evaluated and were categorised into metastatic and non-metastatic nodes showing four different patterns: lymphocyte predominance (LP) pattern, germinal centre predominance (GCP) pattern, sinus histiocytosis (SH) pattern and lymphocyte-depleted pattern. Results: The most frequently encountered pattern of lymph nodes was of LP (436 nodes), followed by GCP (61 nodes), SH (36 nodes) and lymphocyte depleted (18 nodes), respectively. Out of 595, 120 nodes were positive for metastasis, indicating that as the stage advances, the GCP type predominates (the predominant immunomorphological pattern shifts from LP to GCP), facilitating metastasis. Conclusion: Immunomorphological changes in the RLns of OSCCs seem to be pivotal factors that serve as an important prognostic indicator. Whereas the cell-mediated immune response represented by LP and SH appears to resist the metastasis process, the humoral response reflected by GCP favours metastasis.

Species-specific or para-specific antivenoms? On the immunological reactivity and neutralization efficacy for Saw-scaled viper venoms

Species-specific or para-specific antivenoms? On the immunological reactivity and neutralization efficacy for Saw-scaled viper venoms

IgE- and non-IgE-mediated food allergies - an overview

Food allergies are divided into 2 main categories: IgE-mediated and non-IgE-mediated food allergies. Both forms can have significant health effects but differ in mechanism, symptoms, and management. The manifestation of the 2 forms differs between children and adults. These differences can be observed in the prevalence and the type of most common allergens and clinical presentation. The prevalence of food allergies has increased worldwide in recent decades. IgE-mediated allergies are the best researched and documented. They are particularly common in children, while non-IgE-mediated allergies are less well understood and diagnosed, leading to uncertainty about their prevalence. They often manifest as gastrointestinal symptoms that can occur hours to days after ingestion and are often difficult to distinguish from other food intolerances. The occurrence of food allergies varies significantly geographically. Differences in dietary habits, food composition, and environmental factors can partly explain these differences. There are also indications that genetics may play a role. IgE-mediated and non-IgE-mediated food allergies represent a significant and growing challenge for the global healthcare system. This article provides an in-depth review of both types of food allergy, discussing their potential causes, diagnostic possibilities, and available therapeutic strategies. Some diseases represent a mixed form of IgE and non-IgE-mediated immunological adverse reactions. Eosinophilic oesophagitis is the most common eosinophilic disease, and the diagnosis and treatment options are explained in more detail below.

A Novel Monoclonal Antibody against PD-1 for the Treatment of Viral Oncogene-Induced Tumors or Other Cancer.

Programmed cell death 1 (PD-1) and programmed death-ligand 1 (PD-L1) interact to form an immune checkpoint fostering viral infection and viral oncogene-induced tumorigenesis. We generated a novel anti-human PD-1, humanized monoclonal antibody P1801 and investigated its pharmacologic, pharmacokinetic (PK), and pharmacodynamic properties. In vitro binding assays revealed that P1801 uniquely binds to human PD-1 and inhibits its interaction with PD-L1/2. It showed a minor effect on the induction of antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC). P1801 significantly induced the release of IL-2 from activated T-cells but not from nonactivated T-cells. A dose-dependent linear PK profile was observed for the cynomolgus monkeys treated with repeated doses of P1801 at 5 mg/kg to 200 mg/kg once weekly. A four-week repeat-dose toxicity study revealed that P1801 given weekly was safe and well tolerated at doses ranging from 5 to 200 mg/kg/dose. No pathological abnormalities were noted. In humanized PD-1 mice harboring human PD-L1-expressing colon tumor cells, P1801 administered intraperitoneally twice per week at 12 mg/kg significantly inhibited tumor growth and prolonged mouse survival. P1801 displayed unique binding properties different from pembrolizumab and nivolumab. Therefore, it showed distinctive immunological reactions and significant antitumor activities. We are initiating a Phase 1 clinical study to test its combination use with ropeginterferon alfa-2b, which also has antiviral and antitumor activities, for the treatment of cancer.

Evaluating Autoimmune Markers in Relation to Gastrointestinal Measures in Patients With Symptoms of Gastroparesis.

Patients with symptoms (Sx) of gastroparesis (Gp) may have signs of autoimmune disease. We hypothesized that serum autoantibodies in Gp Sx patients are associated with follow-up at 48 weeks (about 11mo) and later autoimmune questionnaires (AQ). One hundred eleven patients: 24 male, 87 female, mean age 43 years, with Gp Sx: 27 diabetic (DM), 78 idiopathic (ID), 6 postsurgical (PS) were studied at baseline, 48 weeks (about 11mo) and long term. Serum western blot (WB) was performed by an enzyme linked immunosorbent assay for qualitative evaluation of serum antibodies to various specific nuclear antigens. A blotting score (GIBS) was used to compare global immunologic reactions. At long-term follow-up (>5y), 38 AQ-responding patients (34 with baseline WB data) were compared with WB by AQ responses. A total of 69/111 (62%) had gastric emptying delay. Correlations exist between WB autoantibodies and clinical follow-up at 48 weeks (about 11mo). 94/96 (97.9%) were WB positive and 62/96 patients had 48-week data. From 5 years follow-up of 38 patients, AQ Sx responses were recurring abdominal pain (89%), lack of energy (87%), exhaustion (74%), and constipation (68%). From AQ and WB data (34 patients), Anti-SSA was higher in those indicating dry mouth and mouth and nose sores and SM was higher in dysphagia (P=0.01 for both). Conclusions In patients with gastroparesis symptoms baseline western blot has significant associations with a number of clinical findings and autoimmune disorders symptoms. These associations suggest possible undiagnosed autoimmune disorders. Further work with Western blotting and other autoimmune measures in patients with gastroparesis symptoms is needed.

Update on the Mechanism of Action of Intravesical BCG Therapy to Treat Non-Muscle-Invasive Bladder Cancer.

While more than four decades have elapsed since intravesical Bacillus Calmette-Guérin (BCG) was first used to manage non-muscle invasive bladder cancer (NMIBC), its precise mechanism of anti-tumor action remains incompletely understood. Besides the classic theory that BCG induces local (within the bladder) innate and adaptive immunity through interaction with multiple immune cells, three new concepts have emerged in the past few years that help explain the variable response to BCG therapy between patients. First, BCG has been found to directly interact and become internalized within cancer cells, inducing them to act as antigen-presenting cells (APCs) for T-cells while releasing multiple cytokines. Second, BCG has a direct cytotoxic effect on cancer cells by inducing apoptosis through caspase-dependent pathways, causing cell cycle arrest, releasing proteases from mitochondria, and inducing reactive oxygen species-mediated cell injury. Third, BCG can increase the expression of programmed death ligand 1 (PD-L1) on both cancer and infiltrating inflammatory cells to impair the cell-mediated immune response. Current data has shown that high-grade recurrence after BCG therapy is related to CD8+ T-cell anergy or 'exhaustion'. High-field cancerization and subsequently higher neoantigen presentation to T-cells are also associated with this anergy. This may explain why BCG therapy stops working after a certain time in many patients. This review summarizes the detailed immunologic reactions associated with BCG therapy and the role of immune cell subsets in this process. Moreover, this improved mechanistic understanding suggests new strategies for enhancing the anti-tumor efficacy of BCG for future clinical benefit.

The Human Monkeypox Virus and Host Immunity: Emerging Diagnostic and Therapeutic Challenges.

This article explores the Human Monkeypox Virus (MPV), a contagious virus that causes disease in both vertebrates and insects. It originated in Denmark in 1958 and expanded beyond Africa during the 1970s. The virus was initially detected in the United States in 2003 following the hospitalisation of a toddler who had been bitten by a prairie dog. The article examines the identification of the virus, its categorization into two genetic groups with dif-ferent levels of harmfulness, and its genetic changes over time due to specific influences. Additionally, it investigates the immunological reaction to MPXV, encompassing both the innate and adaptive systems. The essay also addresses the diagnostic difficulties presented by MPXV's resemblance to other orthopoxviruses and the progress made in molecular diagnos-tics. The paper analyses different therapeutic interventions, such as tecovirimat, an antiviral medication, and JYNNEOS, a vaccine, in terms of their efficacy, potential drawbacks, and the difficulties encountered in managing outbreaks. The future outlook emphasises the ne-cessity of inventive research methodologies, worldwide monitoring, and individualised med-ical treatments to counteract the dissemination of MPXV and alleviate its consequences on public health.

Effect of interval hypoxytherapy on the immune status of hypertensive patients

In recent years, the role of sluggish nonspecific inflammation in the pathogenesis of hypertension has been proven. Oxidative stress that develops in hypertension leads to damage of endothelial glycocalyx with impaired barrier and adaptive functions of endothelium. Activation of transcription factor NF-kB leads to increased synthesis of free oxygen radicals, adhesion molecules (VCAM-1, ICAM-1, P-selectin, E-selectin) and proinflammatory cytokines (TNFα, IL-1, IL-6), triggering the inflammatory process in the endothelium. Modern antihypertensive drugs, affecting various pathogenetic mechanisms of arterial hypertension development, do not fully affect the immune component of arterial hypertension. Therefore, the search for various methods affecting the immunologic reactivity of hypertensive patients remains relevant in our time. The aim of the study was to reveal the effect of interval hypoxytherapy on the state of immunologic reactivity of patients with hypertension stage I. One hundred seventy male patients of 30-45 years old (mean age 38.36±1.64 years) with hypertension stage I (n = 170) who underwent combined treatment including drug therapy and normobaric interval hypoxic therapy in hypoxia-normoxia mode (oxygen content in hypoxic gas mixture was 20.9%) were examined. Indices of redox status and immunologic reactivity before and after interval hypoxytherapy were studied. Having a significant antioxidant effect, interval hypoxytherapy led to the subsidence of oxidative stress, which was indicated by an increase in the activity of superoxide dismutase and glutathione peroxidase in blood erythrocytes and a decrease in the content of malonic dialdehyde in blood as a result of a decrease in the generation of free oxygen radicals and suppression of lipid peroxidation processes. Interval hypoxytherapy resulted in a statistically significant increase in the initially decreased content of CD3+T lymphocytes (p 0.05), CD3+CD4+T lymphocytes (p 0.02), CD3+CD8+T lymphocytes (p 0.02). Initially increased number of B-lymphocytes CD19+, CD16+ lymphocytes, CD95+ lymphocytes significantly decreased. An important result of interval hypoxytherapy was a pronounced anti-inflammatory effect: statistically significant decrease in the content of pro-inflammatory interleukins IL-1β (p 0.02), IL-6 (p 0.01) and increase in the content of anti-inflammatory cytokines IL-4 (p 0.005) and IL-10 (p 0.005), which led to the subsidence of sluggish nonspecific inflammation. Suppression of oxidative stress and normalization of immunological reactivity of hypertensive patients after interval hypoxytherapy led to the subsidence of sluggish nonspecific inflammation, reduction of endothelial dysfunction and restoration of the structure and tone of the vascular wall with a decrease in blood pressure and improvement of the clinical course of hypertension.

Concept and role of immunomodulatory medicinal plants used in Ayurvedic health care system

The statement “prevention is better than cure” and its applications is not a new for Ayurvedic health care system. The statement defines the aim and objectives of Ayurvedic health care system in a nutshell “swasthasya swasthya rakshanam aaturasya vikara prashamanam”. The immune system is certainly considered one of our most discreet biological systems withinside the body. Immunizations may be obtained by us actively or passively. Active immunization includes stimulating with an antigen to generate immunological defenses against a destiny exposure, rather than passive vaccination, which includes administering antibodies to someone who has already been exposed to an antigen. Both plant and animal sources produce immunomodulatory effects to boost the body’s immunological reactivity against infections by engaging the non-specific immune system. Herbal immunomodulators are substances that either activate or suppress innate and adaptive immune responses in the body. Different ailments including allergies, asthma, rheumatoid arthritis, cancer, and other infectious disorders are caused by immune system failure. Therefore, regulating the many infectious illnesses requires significant immune response modification. Global scientific study is currently focused on how various medicinal plant components might alter the immune system. Numerous Indian and “Rasayana” and medicinal plants may have immunomodulatory characteristics. Some of these plants are Tinospora cordifolia, Morus alba, Acacia catechu, Allium sativum, and Mangifera indica. There are many more that are still undiscovered and offer space for greater study.

Advancing Precision in Vascularized Composite Allografts through Multiplex Sensing of Predictive Biomarkers

Vascularized Composite Allotransplantation (VCA) is a promising option for extensive tissue reconstruction. Successful graft outcomes in VCA surgeries require close monitoring of immunological reactions and the progression of ischemic conditions. Excessive immunosuppression may lead to chronic infections and dangerous side effects, while inadequate immunosuppression can result in acute or chronic rejection episodes, jeopardizing graft function. Monitoring the immunological status of an allograft becomes critical when exploring innovative tolerance induction protocols or transitioning patients from conventional therapies. Currently, confirming clinical rejection involves invasive biopsies, which are suboptimal for routine monitoring due to associated risks. Thus, developing non-invasive technologies capable of detecting changes in the immunological status of the graft before apparent clinical signs of inflammation and tissue damage become imperative. To address these limitations, this study proposes a low-cost, portable, flexible, miniaturized, and site-of-care (SOC) platform for multiplex sensing of predictive biomarkers (e.g., interleukin 6 (IL6), lactate) and pH to detect instances of VCA rejection before apparent clinical signs of immunogenic reactions such as inflammation and tissue damage.The sensor fabrication was performed on in-house fabricated porous laser-engraved graphene (PLEG)-electrode, utilizing the laser-induced carbonization (LIC) technique on a thin (12.5 µm) polyimide (PI) surface at optimized laser parameters (laser power-22%, speed-5.2%, and gas flow-50%). The PI sheet was laminated on a polyethylene terephthalate (PET) sheet with silicone adhesive in the middle, helping to dissipate the heat generated during the carbonization process. Compared to conventional screen-printed platforms, the PLEG electrode provides higher surface area, receptor loading, electrical conductivity, ease of surface modification, and improved sensitivity with an average resistance of 40 ± 3 Ω/cm. Subsequently, the connection pad and reference electrodes (RE) were coated with silver/silver chloride (Ag/AgCl) conductive paste for improved connection. The RE was stabilized by coating it with polyvinyl butyrate (PVB) and sodium chloride polymeric cocktail. Three working electrodes (WE) were designated as WE-1 (for IL6), WE-2 (for lactate), and WE-3 (for pH). The sensor construction was initiated with surface activation WEs in 0.50 M H2SO4 containing 0.10 M KCl as a supporting electrolyte by sweeping potential between 1.0 and -1.2 V vs. Ag/AgCl reference electrode. For IL-6 sensing, the WEs were incubated with freshly prepared 1-pyrenebutyric acid N-hydroxysuccinimide (PBASE) as a non-covalent crosslinker (10.0 mM) in dimethylformamide (DMF) for 3 hrs., followed by buffer wash (3 times) to remove any unbound crosslinker. Later, a monoclonal antibody to IL6 was immobilized using peptide bond formation between PBASE and the recognition element, passivated by incubating in Bovine serum albumin (BSA) to block non-specific binding. Upon binding, the antigen-antibody complex's formation results in a decrease in voltammetric signal of the redox signal, generating an IL6-dependent change in current. For lactate detection, an amperometric method utilizes a two-electrode electrochemical setup. The WE comprise a composite Glutaraldehyde/BSA/Lactate-Oxidase/Prussian Blue/PLEG-electrode (GA/BSA/LOx/PB/ PLEG). An Ag/AgCl-coated electrode serves as the RE, and counter electrodes (CE). For the pH sensor (PANI/PLEG), polyaniline (PANI) was electrodeposited at 0.80 V for 600 sec on an activated PLEG electrode washed with distilled water and coated with Nafion (0.25% v/v). For pH measurements, the open-circuit potential (OCP) was measured between the PANI/PLEG electrode and the RE.Post characterization, the sensor’s electrochemical measurements to detect IL6, lactate, and pH were performed in buffer, artificial interstitial fluid, porcine blood, and serum samples with portable potentiostat. Under optimal experimental conditions, the devised platform showed a sensitivity of 2.59 µA/(log10IL6), 0.021 µA/mM, and ~49.39 ± 2 mV in spiked serum for IL6, lactate, and pH, respectively. Subsequently, serum samples of porcine VCA recipients were collected on postoperative days (POD) 0, 2, and 6 and tested using the IL-6, lactate, and pH sensors, respectively. The sensors can successfully track the change in the levels of IL6 (increasing) and lactate (increasing) and the change in pH post-surgery and align with clinical observations of inflammation and ischemia post-allograft on POD6. The sensor results were also validated using commercial IL6 ELISA and lactate kits using the same samples and showed a correlation (r) of r = 0.9885 (n=3, for IL6) and r = 0.9896 (n=3, for lactate). Our results demonstrate that the developed miniaturized platform offers a portable, low-cost, robust, and multiplex sensing platform capable of quantifying predictive biomarkers post-VCA surgeries. This platform presents a promising, miniaturized, and cost-effective solution for the real-time monitoring of biomarkers in VCA models. KEYWORDS: Vascularized Composite Allotransplantation (VCA), Ischemia, Interleukin-6, Lactate, Electrochemical sensor, Porous laser engraved graphene, Multiplex sensing

Skin Telocytes Could Fundament the Cellular Mechanisms of Wound Healing in Platelet-Rich Plasma Administration.

For more than 40 years, autologous platelet concentrates have been used in clinical medicine. Since the first formula used, namely platelet-rich plasma (PRP), other platelet concentrates have been experimented with, including platelet-rich fibrin and concentrated growth factor. Platelet concentrates have three standard characteristics: they act as scaffolds, they serve as a source of growth factors and cytokines, and they contain live cells. PRP has become extensively used in regenerative medicine for the successful treatment of a variety of clinical (non-)dermatological conditions like alopecies, acne scars, skin burns, skin ulcers, muscle, cartilage, and bone repair, and as an adjuvant in post-surgery wound healing, with obvious benefits in terms of functionality and aesthetic recovery of affected tissues/organs. These indications were well documented, and a large amount of evidence has already been published supporting the efficacy of this method. The primordial principle behind minimally invasive PRP treatments is the usage of the patient's own platelets. The benefits of the autologous transplantation of thrombocytes are significant, representing a fast and economic method that requires only basic equipment and training, and it is biocompatible, thus being a low risk for the patient (infection and immunological reactions can be virtually disregarded). Usually, the structural benefits of applying PRP are attributed to fibroblasts only, as they are considered the most numerous cell population within the interstitium. However, this apparent simplistic explanation is still eluding those different types of interstitial cells (distinct from fibroblasts) that are residing within stromal tissue, e.g., telocytes (TCs). Moreover, dermal TCs have an already documented potential in angiogenesis (extra-cutaneous, but also within skin), and their implication in skin recovery in a few dermatological conditions was attested and described ultrastructurally and immunophenotypically. Interestingly, PRP biochemically consists of a series of growth factors, cytokines, and other molecules, to which TCs have also proven to have a positive expression. Thus, it is attractive to hypothesize and to document any tissular collaboration between cutaneous administered PRP and local dermal TCs in skin recovery/repair/regeneration. Therefore, TCs could be perceived as the missing link necessary to provide a solid explanation of the good results achieved by administering PRP in skin-repairing processes.