Platelet and Monocyte Microvesicles as Potential Biomarkers of COVID-19 Severity: A Cross-Sectional Analysis.

Background: Coronavirus disease (COVID-19) induces inflammation, coagulopathy following platelet and monocyte activation, and fibrinolysis, resulting in elevated D-dimer levels. Activated platelets and monocytes produce microvesicles (MVs). We analyzed the differences in platelet and monocyte MV counts in mild, moderate, and severe COVID-19, as well as their correlation with D-dimer levels. Methods: In this cross-sectional study, blood specimens were collected from 90 COVID-19 patients and analyzed for D-dimers using SYSMEX CS-2500. Platelet MVs (PMVs; PMVCD42b+ and PMVCD41a+), monocyte MVs (MMVs; MMVCD14+), and phosphatidylserine-binding annexin V (PS, AnnV+) were analyzed using a BD FACSCalibur instrument. Results: PMV and MMV counts were significantly increased in COVID-19 patients. AnnV+ PMVCD42b+ and AnnV+ PMVCD41a+ cell counts were higher in patients with severe COVID-19 than in those with moderate clinical symptoms. The median (range) of AnnV+ PMVCD42b+ (MV/µL) in mild, moderate, and severe COVID-19 was 1,118.3 (328.1-1,910.5), 937.4 (311.4-2,909.5), and 1,298.8 (458.2-9,703.5), respectively (P =0.009). The median (range) for AnnV+ PMVCD41a+ (MV/µL) in mild, moderate, and severe disease was 885.5 (346.3-1,682.7), 663.5 (233.8-2,081.5), and 1,146.3 (333.3-10,296.6), respectively (P =0.007). D-dimer levels (ng/mL) weak correlated with AnnV+ PMVCD41a+ (P =0.047, r=0.258). Conclusions: PMV PMVCD42b+ and PMVCD41a+ counts were significantly increased in patients with severe clinical symptoms, and PMVCD41a+ counts correlated with D-dimer levels. Therefore, MV counts can be used as a potential biomarker of COVID-19 severity.

The Positive Effect of Four-Week Combined Aerobic-Resistance Training on Body Composition and Adipokine Levels in Obese Females.

Obesity is a metabolic disease that is caused by a lack of physical activity and is associated with an increased risk of chronic inflammation. A total of 40 obese adolescent females with an average age of 21.93 ± 1.35 years and average body mass index (BMI) of 30.81 ± 3.54 kg/m2 were enrolled in this study, randomized, and divided into four groups, i.e., control (CTL; n = 10), moderate intensity aerobic training (MAT; n = 10), moderate intensity resistance training (MRT; n = 10), and moderate intensity combined aerobic-resistance training (MCT; n = 10). The enzyme-linked immunosorbent assay (ELISA) kits method was used to analyze the adiponectin and leptin levels between pre-intervention and post-intervention. Statistical analysis was conducted using a paired sample t-test, while correlation analysis between variables used the Pearson product-moment correlation test. Research data showed that MAT, MRT, and MCT significantly increased adiponectin levels and decreased leptin levels compared to the CTL (p ≤ 0.05). The results of the correlation analysis of delta (∆) data showed that an increase in adiponectin levels was significantly negatively correlated with a decrease in body weight (BW) (r = -0.671, p ≤ 0.001), BMI (r = -0.665, p ≤ 0.001), and fat mass (FM) (r = -0.694, p ≤ 0.001) and positively correlated with an increase in skeletal muscle mass (SMM) (r = 0.693, p ≤ 0.001). Whereas, a decrease in leptin levels was significantly positively correlated with a decrease in BW (r = 0.744, p ≤ 0.001), BMI (r = 0.744, p ≤ 0.001), and FM (r = 0.718, p ≤ 0.001) and negatively correlated with an increase in SMM (r = -0.743, p ≤ 0.001). In summary, it can be concluded that our data show that adiponectin levels increased and leptin levels decreased after the intervention of aerobic, resistance, and combined aerobic-resistance training.

The Comparison of Monocyte Human Leukocyte Antigen-D-Related (mHLA-DR) Expression Levels Between Corona Virus Disease 2019 (COVID-19) Patients and Healthy Subjects.

BACKGROUND: SARS-CoV-2 can trigger a dysfunctional immune response in COVID-19 patients and lead to immunosuppression. HLA-DR molecule expressed on the surface of monocytes, known as mHLA-DR, has been widely used as a reliable marker of immunosuppression. Downregulation of mHLA-DR reflects an immunosuppressed state. This study aimed to compare the expression level of mHLA-DR between COVID-19 patients and healthy subjects concerning immune system dysregulation that can be triggered by SARS-CoV-2 and lead to immunosuppression. METHODS: This was an analytic observational study with a cross-sectional design that measured the mHLA-DR expression in EDTA blood samples from 34 COVID-19 patients and 15 healthy subjects using the BD FACSLyricTM Flow Cytometry System. The mHLA-DR examination results were expressed in AB/C (antibodies bound per cell) that were quantified using a standard curve constructed with Quantibrite phycoerythrin beads (BD Biosciences). RESULTS: Expression of mHLA-DR in COVID-19 patients (n = 34) were 21,201 [2,646-92,384] AB/C, with 40,543.5 [9,797-92,384] AB/C mild cases (n = 22), 21,201 [9,831-31,930] AB/C moderate cases (n = 6), and 7,496 [2,646-13,674] AB/C severe to critical cases (n= 6). Expression of mHLA-DR in healthy subjects (n = 15) was 43,161 [25,147-89,846] AB/C. Based on the Mann-Whitney U test, the mHLA-DR expression in COVID-19 patients significantly differed from the mHLA-DR expression in healthy subjects (p = 0.010). CONCLUSION: The level of mHLA-DR expression in COVID-19 patients was lower and significantly different from healthy subjects. Moreover, immunosuppression could be indicated by the decrease of mHLA-DR expression, which was below the reference range found in severe to critically ill COVID-19 patients.

Acute Limb Ischemia due to Arterial Thrombosis in a Patient with COVID-19 Pneumonia: A Case Report.

The COVID-19 pandemic has caused more than 4 million deaths worldwide to date. During the course of the COVID-19 pandemic, thrombotic complications due to hypercoagulable state have emerged as an important issue. Acute limb ischemia is one of emergency cases in vascular disease caused by a sudden decrease in arterial limbs perfusion. Here, we report a 53-year-old male patient with severe COVID-19 and a history of uncontrolled type 2 diabetes mellitus (T2DM) who developed extensive arterial thrombosis and limb ischemia despite being on therapeutic-dose anticoagulation, requiring surgical intervention. Right and left leg open thrombectomy was performed at day 7 after admission due to the excruciating pain and the worsening of the limb conditions. The patient was transferred to intensive care unit in emergency room because of the unstable hemodynamic and passed away a few hours after the surgery. For critically ill patients with COVID-19, special attention should be paid to abnormal coagulation dysfunction and microcirculatory disorders.

Analysis of Common Beta-Thalassemia (ß-Thalassemia) Mutations in East Java, Indonesia.

Background: The frequency of the beta-thalassemia (ß-thalassemia) gene in Indonesia ranges from 3 to 10%. However, in the East Java province, there is still limited information on the prevalence of ß-thalassemia mutations in clinically diagnosed beta-thalassemia patients of East Java. Therefore, this study aimed to characterize ß-thalassemia mutations in selected patients in the East Java province of Indonesia. Methods: This is an analytical observational study. Diagnosis of ß-thalassemia was based on clinical presentation, complete blood count (CBC), and hemoglobin (Hb) electrophoresis. Blood specimens taken from each patient in three ethylenediaminetetraacetic acid (EDTA) tubes were analyzed for CBC and Hb electrophoresis and processed for DNA extraction and subsequent polymerase chain reaction (PCR). Detection of mutations in Hemoglobin Subunit Beta (HBB) gene exons 1-3 of the ß-thalassemia gene as the common mutation in Indonesia was done using PCR followed by Sanger sequencing. Results: In total, 33 (n = 33) participants were involved in this study with ages ranging from 5 to 17 years comprising 19 women and 14 men. Their ethnic origins were Javanese (n = 30) and Chinese (n = 3). CBC results showed that mean ± standard deviation (SD) for Hb, red blood cell (RBC), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), and red cell distribution width (RDW)-CV were 81.2 ± 7.0 g/L; 3.40 ± 0.39 × 109/L; 71.05 ± 5.72 fL; 24.12 ± 2.45 pg; 33.91 ± 1.47 g/dl; 24.38 ± 6.02%, respectively. Hb electrophoresis revealed that 5 out of 33 participants had beta-thalassemia and 28 out of 33 participants had hemoglobinopathy (Hb) E/beta-thalassemia. Results of Sanger sequencing showed the following genotype variations in the samples: 12 (36.4%) with ß CD26 /ß IVS-I-5; 6 (18.2%) with ß CD26 /ß CD35; 3 (9.1%) with ß CD26 /ß IVS-I-2; 2 (6.1%) with ß CD27/28 /ß CD40; 2 (6.1%) with ß IVS-I-1 /ß CAP+1; and ß CD26 /ß IVS-I-1; ß IVS-I-5 /ß CAP+1; ß IVS-I-5 /ß CD35; ß CD26 /ß CD37; ß CD26 /ß CD15; ß CD26 /ß CD40; and ß IVS-I-5 /ß CD19 in 1 (3%) sample, respectively, and 1 (3%) had no abnormality detected in sequencing even though electrophoresis showed abnormality in the migration pattern. The ß CD26 /ß IVS-I-5 mutation was found in samples that were noted to have Hb E/beta-thalassemia on Hb electrophoresis. Conclusion: The underlying genetic variations are heterogeneous in thalassemia patients in East Java, where 12 variants were found. The most common variant was ß CD26 /ß IVS-I-5, which all accounted for Hb E/beta-thalassemia on Hb electrophoresis. Furthermore, 28 out of 33 participants had hemoglobinopathy (Hb) E/beta-thalassemia.

Central resources of variant discovery and annotation and its role in precision medicine.

Rapid technological advancement in high-throughput genomics, microarray, and deep sequencing technologies has accelerated the possibility of more complex precision medicine research using large amounts of heterogeneous health-related data from patients, including genomic variants. Genomic variants can be identified and annotated based on the reference human genome either within the sequence as a whole or in a putative functional genomic element. The American College of Medical Genetics and Genomics (ACMG) and the Association for Molecular Pathology (AMP) mutually created standards and guidelines for the appraisal of proof to expand consistency and straightforwardness in clinical variation interpretations. Various efforts toward precision medicine have been facilitated by many national and international public databases that classify and annotate genomic variation. In the present study, several resources are highlighted with recognition and data spreading of clinically important genetic variations.

Acquired Hemophilia A Associated with NSAID: A Case Report.

Acquired hemophilia A (AHA) is a blood clotting disorder caused by the presence of autoantibodies (inhibitors) against factor VIII. The typical symptom of this disorder is bleeding under the skin and soft tissue (rarely in the joints), with no family or personal previous history of bleeding. This case reports is tended to build up awareness for better diagnosis and therapy. Woman, 39 years old, bruises on both forearms are intermittent for 2 months with a history of long term drug consumption for headache treatment. Hemostatic test showed the elongation of activated partial thromboplastin test (APTT) to 87.1 (normal 24.4-44.4 seconds) and the decreament of factor VIII (FVIII) activity to 5% (normal 60-150%). Provision of recombinant factor VIII lowered factor VIII activity to 2%. Factor VIII inhibitor titer was 21.12 BU and diagnosis AHA was made. Inhibitor eradication by methylprednisolone tablet 3x16mg which was given for 2 months, improved the APPT to 46.7 seconds and factor VIII activity to 36%. Acquired Hemophilia A should be suspected in an adult bleeding patient with history of taking a long time non-steroidal anti-inflammatory drugs (NSAIDs). This case is a rare case in Indonesia and therefore the procedure for diagnosis needs to be improved in order to avoid errors in delivering a therapy which can cause the decreament of factor VIII activity.

The effects of hemolysis on plasma prothrombin time and activated partial thromboplastin time tests using photo-optical method.

Hemolysis is the most common reason why coagulation test samples are rejected. However, the effects of hemolysis on plasma prothrombin time (PPT) and activated partial thromboplastin time (APTT) are rarely investigated and the results are controversial. This research aims to analyze the effects of hemolysis on PPT and APPT using the photo-optical method.Nonhemolyzed citrate blood samples (n = 30) with normal PPT and APTT underwent 2-step mechanical lysis and then hemoglobin level measurement was carried out at each step. The first lysis was mild to moderate resulting in a hemoglobin level of <0.8 g/dL. These samples were labeled as group 1. The second step showed more severe lysis, which resulted in a plasma hemoglobin level of ≥0.8 g/dL. These samples were labeled as group 2. Analysis was carried out on the PPT and APTT differences between the 2 groups and baseline, as well as between group 1 and group 2 using repeated-measures analysis of variance (ANOVA). The effects of hemolysis were analyzed using linear regression. Receiver operating characteristic (ROC) curve analysis was performed to determine the cut-off value in PPT and APTT.Significantly shorter APTT was measured for group 1 than baseline, (P = .000), group 2 than baseline (P = .000), and group 2 than group 1 (P = .003). With regard to PPT results, those for group 1 were significant shorter than baseline (P = .002), while those for group 2 were significantly longer than group 1 (P = .000). In the correlation assay, the level of hemolysis revealed a mildly significant correlation to APTT (R = 0.245; P = .02). Cut-off value for PPT was 1.55 g/dL (100% sensitivity and 87.9% specificity), while the value for APTT was 0.95 g/dL (75% sensitivity and 62.5% specificity).Not all hemolyzed samples should be rejected for PPT and APTT tests using photo-optical methods.