How to Avoid False-Negative and False-Positive COVID-19 PCR Testing

Background: Up to 40% of test results for COVID-19 in the presence of clinical manifestations of the disease might be negative. The reason for a false-negative result might originate from any step of the analysis: poor-quality or empty swab, poor RNA isolation, inactivation of reverse transcriptase or Taq polymerase in the test. Methods: Here we describe a PCR approach for SARS-CoV-2 detection with swab quality and integrity controlled by human ABL1 mRNA amplification. Designed primers work with the cDNA of the ABL1 gene, not genomic DNA. Results: The simultaneous appearance of three signals corresponding to the nucleocapsid, spike, and ABL1 gene indicates infection with the Omicron strain. The amplification of ABL1 gene and nucleocapsid only indicate other than Omicron infection. The appearance of ABL1 amplification only indicates a true negative result for SARS-CoV-2. All other variants are null and void. Conclusions: A system has been developed for multiplex PCR diagnostics of SARS-CoV-2, which makes it possible to eliminate errors leading to false-negative and false-positive results at all stages of analysis. This is accomplished by the presence of specific primers for human RNA, controlling proper swab application, handling, and all the stages of RT-PCR

Features of Microstructure and Texture Formation of Large-Sized Blocks of C11000 Copper Produced by Electron Beam Wire-Feed Additive Technology

The paper investigated the possibility of obtaining large-sized blocks of C11000 copper on stainless steel substrates via electron beam wire-feed additive technology. The features of the microstructure and grain texture formation and their influence on the mechanical properties and anisotropy were revealed. A strategy of printing large-sized C11000 copper was determined, which consists of perimeter formation followed by the filling of the internal layer volume. This allows us to avoid the formation of defects in the form of drops, underflows and macrogeometry disturbances. It was found that the deposition of the first layers of C11000 copper on a steel substrate results in rapid heat dissipation and the diffusion of steel components (Fe, Cr and Ni) into the C11000 layers, which promotes the formation of equiaxed grains of size 8.94 ± 0.04 μm. As the blocks grow, directional grain growth occurs close to the <101> orientation, whose size reaches 1086.45 ± 57.13 μm. It is shown that the additive growing of large-sized C11000 copper leads to the anisotropy of mechanical properties due to non-uniform grain structure. The tensile strength in the opposite growing direction near the substrate is 394 ± 10 MPa and decreases to 249 ± 10 MPa as the C11000 blocks grows. In the growing direction, the tensile strength is 145 ± 10 MPa

Genetic Lesions in Russian CLL Patients with the Most Common Stereotyped Antigen Receptors

Chronic lymphocytic leukemia (CLL) is one of the most common B-cell malignancies in Western countries. IGHV mutational status is the most important prognostic factor for this disease. CLL is characterized by an extreme narrowing of the IGHV genes repertoire and the existence of subgroups of quasi-identical stereotyped antigenic receptors (SAR). Some of these subgroups have already been identified as independent prognostic factors for CLL. Here, we report the frequencies of TP53, NOTCH1, and SF3B1 gene mutations and chromosomal aberrations assessed by NGS and FISH in 152 CLL patients with the most common SAR in Russia. We noted these lesions to be much more common in patients with certain SAR than average in CLL. The profile of these aberrations differs between the subgroups of SAR, despite the similarity of their structure. For most of these subgroups mutations prevailed in a single gene, except for CLL#5 with all three genes affected by mutations. It should be noted that our data concerning the mutation frequency in some SAR groups differ from that obtained previously, which could be due to the population differences between patient cohorts. The research in this area should be important for better understanding the pathogenesis of CLL and therapy optimization

Clinical Study of the Relationship between Sjögren Syndrome and T-Cell Large Granular Lymphocytic Leukemia: Single-Center Experience

The relationship between Sjögren syndrome (SS) and T-cell large granular lymphocytic (T-LGL) leukemia remains unclear. In this paper, we report for the first time a large case series of 21 patients with primary and secondary SS associated with T-LGL leukemia. Our results suggest the importance of considering T-LGL leukemia in the diagnostic evaluation of SS patients, particularly when neutropenia occurs. We also postulate that elevated antinuclear antibody titers in patients with T-LGL leukemia indicate the need for the clinical assessment of SS. To assess whether SS affects the frequency of the signal transducer and activator of transcription 3 (STAT3) gene mutations in T-LGL leukemia, we examined STAT3 mutations by next-generation sequencing in two cohorts of patients: with SS-associated T-LGL leukemia and T-LGL leukemia in the setting of rheumatic diseases but without SS. While our results suggest that SS, per se, is not associated with an increased frequency of STAT3 mutations in T-LGL leukemia, further studies are needed to better assess the role of the STAT pathway in the development of concomitant SS and T-LGL leukemia

In-Situ Al-Mg Alloy Base Composite Reinforced by Oxides and Intermetallic Compounds Resulted from Decomposition of ZrW₂O₈ during Multipass Friction Stir Processing

In the presented work, the effect of friction stir processing admixing the zirconium tungstate ZrW2O8 powder on the microstructure, mechanical and tribological properties of the AA5056 Al-Mg alloy stir zone has been studied. The FSP resulted in obtaining dense composite stir zones where α-ZrW2O8 underwent the following changes: (i) high-temperature transformation into metastable β’-ZrW2O8 and (ii) decomposition into WO3 and ZrO2 oxides followed by the formation of intermetallic compounds WAl12 and ZrAl3. These precipitates served as reinforcing phases to improve mechanical and tribological characteristics of the obtained fine-grained composites. The reduced values of wear rate and friction coefficient are due to the combined action the Hall–Petch mechanism and reinforcement by the decomposition products, including Al2O3, ZrO2, β’-ZrW2O8 and intermetallic compounds such as WAl12 and ZrAl3. Potential applications of the above-discussed composites maybe related to their improved tribological characteristics, for example in aerospace and vehicle-building industries

Viral Load and Patterns of SARS-CoV-2 Dissemination to the Lungs, Mediastinal Lymph Nodes, and Spleen of Patients with COVID-19 Associated Lymphopenia

Lymphopenia is a frequent hematological manifestation, associated with a severe course of COVID-19, with an insufficiently understood pathogenesis. We present molecular genetic immunohistochemical, and electron microscopic data on SARS-CoV-2 dissemination and viral load (VL) in lungs, mediastinum lymph nodes, and the spleen of 36 patients who died from COVID-19. Lymphopenia <1 × 109/L was observed in 23 of 36 (63.8%) patients. In 12 of 36 cases (33%) SARS-CoV-2 was found in lung tissues only with a median VL of 239 copies (range 18–1952) SARS-CoV-2 cDNA per 100 copies of ABL1. Histomorphological changes corresponding to bronchopneumonia and the proliferative phase of DAD were observed in these cases. SARS-CoV-2 dissemination into the lungs, lymph nodes, and spleen was detected in 23 of 36 patients (58.4%) and was associated with the exudative phase of DAD in most of these cases. The median VL in the lungs was 12,116 copies (range 810–250281), lymph nodes—832 copies (range 96–11586), and spleen—71.5 copies (range 0–2899). SARS-CoV-2 in all cases belonged to the 19A strain. A immunohistochemical study revealed SARS-CoV-2 proteins in pneumocytes, alveolar macrophages, and bronchiolar epithelial cells in lung tissue, sinus histiocytes of lymph nodes, as well as cells of the Billroth pulp cords and spleen capsule. SARS-CoV-2 particles were detected by transmission electron microscopy in the cytoplasm of the endothelial cell, macrophages, and lymphocytes. The infection of lymphocytes with SARS-CoV-2 that we discovered for the first time may indicate a possible link between lymphopenia and SARS-CoV-2-mediated cytotoxic effect