The Diagnostic Utility of Natural Killer Cell Subsets in Deep Vein Thrombosis.

Background: Natural killer (NK) cells are important components of adaptive and innate immune responses. NK cell subsets have different functions and may play a role in vascular disorders. This study aimed to evaluate the proportions of NK cells and their subsets to determine whether they can be used as markers of venous thrombosis and to identify whether there was a link between NK cell proportion and citrullinated histone (H3) levels. Patients and Methods: This study included 100 participants divided into Group I (n=50, patients with deep venous thrombosis (DVT)) and Group II (n=50, age- and sex-matched healthy controls). Group I was further categorized into Group Ia (n=25, patients with acute DVT) and Group Ib (n=25, patients with chronic DVT). The proportions of NK cells and their subsets were evaluated by flow cytometry using CD3/CD16/CD56. The levels of citrullinated histones (H3) were estimated using enzyme-linked immunosorbent assay (ELISA). Results: Compared to the control group, DVT patients had a significantly lower proportion of (CD56 dim/CD16+) NK cells, a significantly higher proportion of (CD56-/CD16+) NK cells and a high level of citrullinated histone (H3). Conclusion: NK cell subsets and citrullinated histone (H3) could be used as markers for DVT and as targets for therapeutic drugs to inhibit the formation or progression of thrombosis.

Prognostic utility of MicroRNA-221 and interleukin-6 in cerebral ischemic stroke.

Cerebral ischemic stroke has a significant mortality rate and persistent impairment. The initial diagnosis of stroke occurs by magnetic resonance imaging and computed tomography. There is a strong need for more accessible, less expensive, and non-invasive methods besides the neuroimaging methods. MicroRNAs (miRNAs) are critical regulators for ischemic stroke as they are involved in stroke pathophysiology. The goal of the current study was to determine whether microRNA-221 (miR-221) could be used as a diagnostic biomarker for patients with ischemic stroke, and whether it can serve as a promising indicator of the disease severity especially if combined with interleukin-6 (IL-6). The study included 90 subjects, 45 cerebral ischemic stroke patients and 45 controls. MiR-221 was evaluated by quantitative real-time polymerase chain reaction (q-PCR) and IL-6 by enzyme-linked immunosorbent assay (ELISA). Our study results revealed that the serum miR-221 level was significantly reduced in cerebral ischemic stroke patients when compared to the control group (p<0.0001). In addition, serum miR-221 showed a significant negative correlation with cerebral stroke severity (p<0.0001), whereas serum IL-6 showed a significant positive correlation with cerebral stroke severity (p < 0.0001). We also analyzed the receiver operator characteristic (ROC) curve and found that area under the ROC curve (AUC) for severity of ischemic stroke by miR-221 was 0.97 (95% confidence intervall0.93-1, p<0.001). Notably, the combination of serum miR-221 with IL-6 for prediction of ischemic stroke severity showed both increased sensitivity/specificity (AUC=0.99, 95% confidence interval 0.96-1, p<0.001) than miR-221 alone. We concluded that miR-221 constituted a non-invasive, sensitive, and specific biomarker that could be used for diagnosis of ischemic stroke and for prediction of its severity.

The Correlation Between C-Peptide and Severity of Peripheral Atherosclerosis in Type 2 Diabetes Mellitus.

Background: One of the major issues affecting global health is Diabetes mellitus (DM), not only in terms of the disease itself but also its complications. Macrovascular complications are both common and serious, affecting many patients. This study aimed to assess fasting C-peptide levels and correlate them with the severity of the peripheral arterial disease complicating type 2 DM (T2DM). Patients and Methods: This study included 200 participants who were categorized into two groups: Group I (n=100, patients with T2DM complicated by femoropopliteal arterial atherosclerosis) and Group II (n=100, healthy age- and sex-matched individuals serving as controls). Fasting C-peptide levels were estimated using an immunochemiluminometric assay. Results: Fasting C-peptide levels were significantly higher in Group I than in the control group. Fasting C-peptide levels were positively correlated with the severity of atherosclerosis. In addition, the receiver operating characteristic (ROC) curve analysis revealed that fasting C-peptide levels served as a specific and sensitive marker for detecting the severity of this disease. Conclusion: Fasting C-peptide levels can be used as a sensitive and specific indicator of the severity of femoropopliteal arteriosclerosis that complicates T2DM.

Impact of Toll-Like Receptor 2 and 9 Gene Polymorphisms on COVID-19: Susceptibility, Severity, and Thrombosis.

Background: Toll-like receptors (TLRs) play an important role in activation of innate and adaptive immune responses. Aim: We aimed to detect the association between TLR2 rs5743708 G>A and TLR9 rs5743836 C>T variants and COVID-19 disease susceptibility, severity, and thrombosis by using neutrophil extracellular traps (NETs). Subjects and Methods: We included 100 adult COVID-19 patients as well as 100 age- and gender-matched normal controls. Participants were genotyped for TLR2 rs5743708 and TLR9 rs5743836. Citrullinated Histone (H3) was detected as an indicator of NETs. Results: The mutant (G/A and C/C) genotypes and (A and C) alleles of TLR2 rs5743708 and TLR9 rs5743836, respectively, have been significantly related to a higher risk of COVID-19 infection, representing a significant risk factor for the severity of COVID-19. There was no significant association between the two variants and citrullinated histone (H3). Conclusion: TLR2 rs5743708 and TLR9 rs5743836 variants have been significantly related to a higher risk and severity of COVID-19 infection but had no effect on thrombus formation.

Expression levels of complement regulatory proteins (CD35, CD55 and CD59) on peripheral blood cells of patients with chronic kidney disease.

BACKGROUND: Altered regulation of the complement system is associated with multiple kidney diseases. CD35, CD55 and CD59 regulate the complement system, and changes in their expression have previously been linked with kidney disease. This study assessed whether changes in the expression levels of these proteins are associated specifically with chronic kidney disease (CKD) to understand its pathogenesis. MATERIALS AND METHODS: Sixty CKD patients and 60 age-matched controls were enrolled and divided into two groups: Group I (n=30 pediatric patients and n=30 controls) and Group II (n=30 adult patients and n=30 controls). The expression of CD35, CD55 and CD59 on peripheral blood cells was evaluated by flow cytometry as the proportion of positive cells expressing the marker and mean fluorescence intensity (MFI), also the relation of these markers to the stage of CKD was also evaluated. RESULTS: Pediatric and adult CKD patients had significantly lower proportion of erythrocytes expressing CD35, CD55 and CD59 than healthy controls (P<0.001). In pediatric CKD patients, there was no significant difference in the three studied markers on neutrophils, lymphocytes and monocytes. The changes in expression of CD35, CD55 and CD59 on leukocytes were more pronounced in adult patients, who had lower proportion of CD59-positive neutrophils, CD35- and CD59-positive lymphocytes, and CD59-positive monocytes, as well as lower expression of CD59 on neutrophils and monocytes than adult controls (P<0.001, P=0.019, P<0.001, P=0.026, P<0.001 and P=0.003, respectively). The eGFR directly correlated with the proportion of positivity of some of those markers on peripheral leukocytes while there was inverse correlation between the disease stage and the same markers. CONCLUSION: There are alterations in the patterns of expression of complement regulatory proteins CD35, CD55 and CD59 on peripheral blood cells of patients with CKD compared with healthy controls.

Platelet-derived growth factor-beta as a new marker of deep venous thrombosis.

BACKGROUND: Deep venous thrombosis (DVT) is associated with significant morbidity and mortality. Thus, there is a great need to demonstrate a more efficient biomarker that would confirm the diagnosis of DVT. Our work aimed to evaluate the role of platelet-derived growth factor-beta (PDGF-B) as a new marker of DVT and its correlation with other radiological and laboratory tools used for the diagnosis. MATERIALS AND METHODS: A case-control study enrolled forty patients selected from our university hospital between April 2018 and August 2018, who divided into two groups: Group I (n = 20) consisted of patients diagnosed with acute venous thrombosis and Group II (n = 20) consisted of patients diagnosed with chronic venous thrombosis. Twenty samples were collected from age- and gender-matched apparently healthy controls to be used as a control. Venous duplex ultrasonography, routine laboratory investigations, D-dimer (DD), and protein expression of PDGF-B were performed on all patients. RESULTS: There was a highly significant increase in a protein expression of PDFG-B in all cases of acute and chronic venous thrombosis compared to the control group with P < 0.001; furthermore, it was more specific than DD for the detection of DVT (specificity 95% and 90%, respectively). CONCLUSION: Our study submits a novel association of PDGF-B plasma levels with DVT, and PDGF-B is considered to be a more specific indicator for DVT than is DD.

Significance of neuropilin-1 expression, ceruloplasmin oxidase and copper in egyptian children with b-lineage acute lymphoblastic leukemia

Background: Neuropilins are transmembrane glycoproteins that act as receptors for vascular endothelial growth factors (VEGF) and are involved in the process of tumor angiogenesis. Ceruloplasmin is a member of the multi copper oxidase family. It has antioxidant properties that play a central role in protection of the body against advanced oxidation protein products.Objective: This study was aimed to assess the expression of Neuropilin-1(NRP-1) on blasts of B-lineage precursor lymphoblastic leukemia (ALL) to be used in the diagnostic panel of this disease. We also aimed to assess the alteration of the levels of ceruloplasmin oxidase and copper as a compensatory mechanism to minimize the effects of reactive oxygen species resulting from leukemias.Subjects and Methods: This study was conducted on 40 children with newly diagnosed B-lineage precursor lymphoblastic leukemia. 40 age-matched controls were enrolled to serve as control. The expression of NRP-1 on peripheral blood samples was evaluated by flow cytometry as the proportion of positive cells expressing the marker. Ceruloplasmin oxidase and copper levels were assessed by immunoturbidimetric assay.Results: There was highly significant increase in the proportion of positivity of NRP -1 in patients compared with control group (P<0.001) Ceruloplasmin oxidase and copper levels were also higher in patients compared with control group (P<0.001).Conclusions: It could be concluded that NRP-1 is a valuable marker for diagnosis of B-lineage ALL. There is an increase in the levels of ceruloplasmin oxidase and copper which at the time of diagnosis of B-ALL