Circulating microRNAs as predictive biomarkers of coronary artery diseases in type 2 diabetes patients

BackgroundType 2 diabetes mellitus (T2DM) is an increasing metabolic disorder mostly resulting from unhealthy lifestyles. T2DM patients are prone to develop heart conditions such as coronary artery disease (CAD) which is a major cause of death in the world. Most clinical symptoms emerge at the advanced stages of CAD; therefore, establishing new biomarkers detectable in the early stages of the disease is crucial to enhance the efficiency of treatment. Recently, a significant body of evidence has shown alteration in miRNA levels associate with dysregulated gene expression occurring in T2DM and CAD, highlighting significance of circulating miRNAs in early detection of CAD arising from T2DM. Therefore, it seems crucial to establish a link between the miRNAs prognosing value and development of CAD in T2DM.AimThis study provides an overview on the alterations of the circulatory miRNAs in T2DM and various CADs and consider the potentials of miRNAs as biomarkers prognosing CADs in T2DM patients.Materials and MethodsLiterature search was conducted for miRNAs involved in development of T2DM and CAD using the following key words: “miRNAs”, “Biomarker”, “Diabetes Mellitus Type 2 (T2DM)”, “coronary artery diseases (CAD)”. Articles written in the English language.ResultThere has been shown a rise in miR‐375, miR‐9, miR‐30a‐5p, miR‐150, miR‐9, miR‐29a, miR‐30d, miR‐34a, miR‐124a, miR‐146a, miR‐27a, and miR‐320a in T2DM; whereas, miR‐126, miR‐21, miR‐103, miR‐28‐3p, miR‐15a, miR‐145, miR‐375, miR‐223 have been shown to decrease. In addition to T2DM, some miRNAs such as mirR‐1, miR‐122, miR‐132, and miR‐133 play a part in development of subclinical aortic atherosclerosis associated with metabolic syndrome. Some miRNAs increase in both T2DM and CAD such as miR‐1, miR‐132, miR‐133, and miR‐373‐3‐p. More interestingly, some of these miRNAs such as miR‐92a elevate years before emerging CAD in T2DM.Conclusiondysregulation of miRNAs plays outstanding roles in development of T2DM and CAD. Also, elevation of some miRNAs such as miR‐92a in T2DM patients can efficiently prognose development of CAD in these patients, so these miRNAs can be used as biomarkers in this regard.     

Non-Thermal Plasma Reduction of Ag+ Ions into Silver Nanoparticles in Open Atmosphere under Statistically Optimized Conditions for Biological and Photocatalytic Applications

An environmentally friendly non-thermal DC plasma reduction route was adopted to reduce Ag⁺ ions at the plasma–liquid interface into silver nanoparticles (AgNPs) under statistically optimized conditions for biological and photocatalytic applications. The efficiency and reactivity of AgNPs were improved by statistically optimizing the reaction parameters with a Box–Behnken Design (BBD). The size of the AgNPs was chosen as a statistical response parameter, while the concentration of the stabilizer, the concentration of the silver salt, and the plasma reaction time were chosen as independent factors. The optimized parameters for the plasma production of AgNPs were estimated using a response surface methodology and a significant model p < 0.05. The AgNPs, prepared under optimized conditions, were characterized and then tested for their antibacterial, antioxidant, and photocatalytic potentials. The optimal conditions for these three activities were 3 mM of stabilizing agent, 5 mM of AgNO₃, and 30 min of reaction time. Having particles size of 19 to 37 nm under optimized conditions, the AgNPs revealed a 82.3% degradation of methyl orange dye under UV light irradiation. The antibacterial response of the optimized AgNPs against S. aureus and E. coli strains revealed inhabitation zones of 15 mm and 12 mm, respectively, which demonstrate an antioxidant activity of 81.2%.     

Rituximab abrogates aquaporin-4–specific germinal center activity in patients with neuromyelitis optica spectrum disorders

Neuromyelitis optica spectrum disorders (NMOSDs) are caused by immunoglobulin G (IgG) autoantibodies directed against the water channel aquaporin-4 (AQP4). In NMOSDs, discrete clinical relapses lead to disability and are robustly prevented by the anti-CD20 therapeutic rituximab; however, its mechanism of action in autoantibody-mediated disorders remains poorly understood. We hypothesized that AQP4-IgG production in germinal centers (GCs) was a core feature of NMOSDs and could be terminated by rituximab. To investigate this directly, deep cervical lymph node (dCLN) aspirates (n = 36) and blood (n = 406) were studied in a total of 63 NMOSD patients. Clinical relapses were associated with AQP4-IgM generation or shifts in AQP4-IgG subclasses (odds ratio = 6.0; range of 3.3 to 10.8; P < 0.0001), features consistent with GC activity. From seven dCLN aspirates of patients not administered rituximab, AQP4-IgGs were detected alongside specific intranodal synthesis of AQP4-IgG. AQP4-reactive B cells were isolated from unmutated naive and mutated memory populations in both blood and dCLNs. After rituximab administration, fewer clinical relapses (annual relapse rate of 0.79 to 0; P < 0.001) were accompanied by marked reductions in both AQP4-IgG (fourfold; P = 0.004) and intranodal B cells (430-fold; P < 0.0001) from 11 dCLNs. Our findings implicate ongoing GC activity as a rituximab-sensitive driver of AQP4 antibody production. They may explain rituximab’s clinical efficacy in several autoantibody-mediated diseases and highlight the potential value of direct GC measurements across autoimmune conditions.

Immunoglobulin G (IgG) autoantibodies directed against the extracellular domain of the water channel aquaporin-4 (AQP4) are directly causative in patients with neuromyelitis optica spectrum disorders (NMOSDs) (1–4). AQP4-IgGs are predominantly of the IgG1 subclass, and their major proposed pathogenic mechanism is via complement-mediated damage to the AQP4-rich astrocyte end feet (5). In NMOSDs, patient disability is accrued through discrete clinical relapses, typically affecting the spinal cord and/or optic nerve (6, 7). However, the immunobiology underlying these attacks is poorly understood, and few serum biomarkers can accurately predict relapses (8).Traditionally, ongoing autoantibody production is considered to occur via two broadly discrete cellular pathways: continual germinal center (GC) activity versus long-lived plasma cells (LLPCs) (9). GCs are specialized microenvironments, typically located within secondary lymphoid organs, where antigen-reactive B cells diversify and mature their immunoglobulin genes via somatic hypermutation, with help from specialized lymphoid-resident T follicular helper (Tfh) cells (10). The process of somatic hypermutation is commonly observed alongside a DNA excision process known as class-switch recombination. Together, somatic hypermutation and class-switch recombination can generate high-affinity IgG responses. Autoantigen reactivity of the B cell receptor (BCR) may either arise de novo following somatic hypermutation in GCs or be originally encoded by antigen-reactive germline BCRs expressed by naive B cells (10, 11). Ongoing GC activity may be responsible for the prolonged presence of autoantibodies, such as AQP4-IgGs (9, 12). In an alternative model, LLPCs that successfully exit active GCs and acquire a bone marrow niche may autonomously persist for decades after an autoimmunizing event. These niched LLPCs are thought to secrete >90% of human serum IgG, including a variety of autoantibodies (13, 14).To date, a series of observations suggest that GC activity may play an important role in AQP4-IgG generation. First, close correlations between serum AQP4-IgG levels and AQP4-IgG secreted in vitro by circulating B cells suggest a limited role for LLPCs in AQP4-IgG generation (12, 15). Second, the detection of circulating AQP4-reactive naive B cells identifies a source of cells that could enter GCs and are reported to share clonal relationships with the hypermutated BCRs of intrathecal AQP4-reactive plasma cells (16, 17). Next, annualized relapse rates (ARR) in NMOSDs are robustly reduced by multiple immunotherapies likely to spare nonproliferative CD20⁻ LLPCs, including the anti-CD20 monoclonal antibody rituximab (RTX) (18–20); however, likely because RTX spares the LLPCs, it does not reduce serum AQP4-IgG levels, an observation that presents a potential clinical–serological paradox in a disease with proven pathogenic autoantibodies (21, 22).We hypothesized that the rapid clinical efficacy of RTX observed in patients with NMOSDs may be explained by its direct disruption of active GC reactions, impacting the most affinity matured, and hence pathogenic, B cells and antibodies. However, contradictory data from both human and mouse studies mean that it remains unclear whether RTX effectively depletes B cells within secondary lymphoid organs (23–25). Further, the putative role of GCs in NMOSDs has not been studied directly. In autoimmune diseases of the central nervous system (CNS), the lymphoid organs that drain meningeal lymphatics represent the most plausible anatomical site of active GCs, the deep cervical lymph nodes (dCLNs) (26).To address these concepts, we studied 63 patients with NMOSDs as a prototypical model of an autoantibody-mediated condition. From patients seen as part of routine clinical practice in two specialist NMO centers, we identified clinical relapses in association with proxy measures of an active GC response: class-switch recombination and de novo AQP4-IgM production. Next, to directly sample the secondary lymphoid organs most likely to generate a GC response to neuronal antigens, we aspirated dCLNs from NMOSD patients. These aspirates contained intranodal AQP4-specific B cells and evidence of local, intranodal AQP4-IgG synthesis, both of which were rapidly and efficiently abrogated by RTX over a timescale consistent with clinical remission. Our data present direct insights into the immunological drivers of NMOSD, highlight the effects of RTX in a model of human autoantibody-mediated illness, and provide a platform for the direct analyses of GCs in human autoimmunity.     

Establishment of Patient-Derived Gastric Cancer Organoid Model From Tissue Obtained by Endoscopic Biopsies

Cancer organoids are three-dimensional mini-organ analogues derived from cancer tissues and have been proposed as models capable of simulating the structure and function of human organs and tissues in vitro. We sought to establish gastric cancer patient-derived organoids (PDOs) from tissues obtained by endoscopic biopsies. Gastric cancer-PDOs were successfully established and cultured from cancer tissues with gastric adenocarcinoma by endoscopic biopsies. To confirm that gastric cancer-PDOs were derived from cancer tissue, the consistency of the original cancer tissue was assessed by histopathological examination. As a result, it was confirmed that the shape and internal structure of gastric cancer-PDO were derived from the original gastric cancer cells, and the tumor specificity of gastric cancer-PDO was confirmed through Periodic acid-Schiff (PAS) and polyclonal carcinoembryonic antigen antibody staining. These results demonstrate that gastric cancer-PDO models show the characteristics of primary tumors and have potential for drug screening and providing a personalized medicine platform.     

Effect of Different Wax Pattern Manufacturing Techniques on the Marginal Fit of Lithium Disilicate Crowns

Purpose: The present study evaluated the marginal gap of lithium disilicate crowns fabricated through three different wax pattern techniques; Conventional, Milling and 3D-printing. Materials and Methods: Thirty stone models were replicated from a stainless-steel model representing a prepared tooth; ten were sent to make conventional wax patterns while the remaining were sent to a digital dental scanner. The computer aided design was completed and STL (Standard Tessellation Language) files were sent to either milling or 3D-printing machines. All wax patterns (n = 30) were pressed, and a stabilizing instrument was used to secure the crowns on the master model. The marginal gap was measured at 18 points for each crown using a digital microscope (µm) (n = 540) and compared using One-way ANOVA (p ≤ 0.05). Results: There was a significant difference in the marginal gap value between all three groups (p < 0.01) where the milled group showed the least mean gap (28.87 ± 30.18 µm), followed by 3D printed (47.85 ± 27.44 µm), while the highest mean marginal gap was found in the conventional group (63.49 ± 28.05 µm). Conclusion: Milled and 3D-printed wax patterns produced better fitting crowns compared to conventional techniques.     

A case of lymphoproliferative disorder of NK‐cells: aggressive immunophenotype but indolent behavior

Distinguishing chronic lymphoproliferative disorder of NK‐cells from aggressive NK‐cell leukemia is critical because they have distinct clinical course and management. Immunophenotyping plays a key role in distinguishing these two entities, however, it could not be used as sole criteria and clinical/laboratory findings are equally important.     

Clinical utility of haematological inflammatory biomarkers in predicting 30-day mortality in hospitalised adult patients with COVID-19

Coronavirus disease 2019 (COVID-19) is a multisystem disease affecting respiratory, cardiovascular, gastrointestinal, neurological, immunological and haematological systems. The most important indices that have been studied are platelet (PLT) indices in addition to the PLT count and red blood cell distribution width (RDW). This retrospective study included 95 patients with COVID-19 and was conducted at the Hospital Isolation, Scientific and Medical Research Centre and Clinical Pathology Department at Zagazig University Hospitals, Egypt over 6 months from March to August 2021. All patients on admission had a full blood count, which included white blood cell (WBC) count, haemoglobin, RDW, PLT count and its indices in addition to PLT-to-WBC ratio (PWR) and PLT-to-lymphocyte ratio (PLR), which were calculated for all the study patients. There were significant linear correlations for higher levels of the PLR, PWR and RDW and mortality rate (p = 0.03, p < 0.001 and p < 0.001 respectively). Moreover, on multivariable analysis the RDW, PLT count and PWR levels were independent prognostic predictors for mortality with a hazard ratio [HR] of 1.25 (95% confidence interval [CI] 1.09–1.44, p = 0.002), 1.00 (95% CI 0.99–1.00, p = 0.03) and 2.3 (95% CI 1.21–4.48, p = 0.01) respectively. The RDW and PLT indices are accessible predictors that can be valuable prognostic factors for survival assessment and risk stratification of COVID-19.     

COVID-19 related vascular complications in a pediatric patient: A case report

The novel coronavirus infection (COVID-19) caused by the SARS-CoV-2 virus is a relatively recent infection that has resulted in a global pandemic, appearing first at the end of 2019. While initially presenting as a predominantly respiratory disease, with a classical picture of fever, dry cough, dyspnea and, in some cases anosmia and ageusia, recent cases have shown increasingly atypical and more systemic manifestation of the disease. A precise understanding of the extent and pathophysiology of COVID-19 remains underway to this day, particularly concerning its behavior in the pediatric population. Moreover, there has been an increasing number of COVID-19 reports with neurological complications and manifestations, prompting inquiry into neuroinvasion. Postulations include indirect invasion through a surge of inflammatory mediators “cytokine storm” and subsequent widespread endothelial injury; and direct neural tropism. We report the case of a previously healthy 12-year-old male presenting with acute right-sided hemiparesis, new-onset seizures and a generalized petechial rash. Laboratory tests revealed elevated inflammatory markers and radiological investigations confirmed an evolving left middle cerebral artery (MCA) infarct and large vessel vasculitis. Testing for SARS-CoV-2 infection was positive.