Increased LAMP1 Expression Enhances SARS-CoV-1 and SARS-CoV-2 Production in Vero-Derived Transgenic Cell Lines.

Coronaviridae is a family of single-stranded RNA (ssRNA) viruses that can cause diseases with high mortality rates. SARS-CoV-1 and MERS-CoV appeared in 2002‒2003 and 2012, respectively. A novel coronavirus, SARS-CoV-2, emerged in 2019 in Wuhan (China) and has caused more than 5 million deaths in worldwide. The entry of SARS-CoV-1 into the cell is due to the interaction of the viral spike (S) protein and the cell protein, angiotensin-converting enzyme 2 (ACE2). After infection, virus assembly occurs in Golgi apparatus-derived vesicles during exocytosis. One of the possible participants in this process is LAMP1 protein. We established transgenic Vero cell lines with increased expression of human LAMP1 gene and evaluated SARS-CoV-1 and SARS-CoV-2 production. An increase in the production of both viruses in LAMP1-expressing cells when compared with Vero cells was observed, especially in the presence of trypsin during infection. From these results it can be assumed that LAMP1 promotes SARS-CoV-1 and SARS-CoV-2 production due to enhanced exocytosis.

[Increased LAMP1 Expression Enhances SARS-CoV-1 and SARS-CoV-2 Production in Vero-Derived Transgenic Cell Lines].

Coronaviridae is a family of single-stranded RNA (ssRNA) viruses that can cause diseases with high mortality rates. SARS-CoV-1 and MERS-CoV appeared in 2002-2003 and 2012, respectively. A novel coronavirus, SARS-CoV-2, emerged in 2019 in Wuhan (China) and has caused more than 5 million deaths in worldwide. The entry of SARS-CoV-1 into the cell is due to the interaction of the viral spike (S) protein and the cell protein, angiotensin-converting enzyme 2 (ACE2). After infection, virus assembly occurs in Golgi apparatus-derived vesicles during exocytosis. One of the possible participants in this process is LAMP1 protein. We established transgenic Vero cell lines with increased expression of human LAMP1 gene and evaluated SARS-CoV-1 and SARS-CoV-2 production. An increase in the production of both viruses in LAMP1-expressing cells when compared with Vero cells was observed, especially in the presence of trypsin during infection. From these results it can be assumed that LAMP1 promotes SARS-CoV-1 and SARS-CoV-2 production due to enhanced exocytosis.

Detection of SARS-CoV-2 RNA in nasopharyngeal swabs from COVID-19 patients and asymptomatic cases of infection by real-time and digital PCR.

In this work we tested two reagent kits developed by us for detecting SARS-CoV-2 RNA using a fragment of the ORF1ab gene in digital PCR and real-time PCR formats. Data were obtained on the detection of SARS-CoV-2 virus RNA in nasopharyngeal swabs of patients with COVID-19 and asymptomatic carriers. The developed reagent kits provided 100% sensitivity and a detection limit of 103 GE / ml for qPCR, and at least 200 copies / ml of viral RNA when performing digital PCR. These methods were tested using a panel of 1,328 samples collected from patients with suspected COVID-19 at the beginning of 2020 in the Russian Federation. It has been shown that dPCR is more sensitive and can be used to analyze samples with low viral load, including those from patients without clinical symptoms. dPCR significantly improves the accuracy of laboratory research and significantly reduces the number of false negative results in the diagnosis of SARS-CoV-2. Determination of the concentration of SARS-CoV-2 RNA in patients with different clinical course of the disease showed that the concentration of viral RNA can sharply decrease in the first days of the disease. A low concentration of viral RNA in samples from patients is also characteristic of asymptomatic disease. Digital PCR provides a higher detection rate for asymptomatic cases, which is approximately 75% of those infected, as opposed to 45% for real-time PCR. The results obtained on the use of the digital PCR method for detecting SARS-CoV-2 RNA showed that this method is especially suitable for detecting RNA in case of its low concentration in contacts, as well as for monitoring changes in viral load in convalescent patients.

[Laparoscopic adhesiolysis and prevention of abdominal adhesions by using mechanical barriers]. / Laparoskopicheskii adgeziolizis i bar'ernaia profilaktika spaechnogo protsessa briushnoi polosti.

Adhesions commonly result from abdominal and pelvic surgical procedures and may result in intestinal obstruction, infertility, chronic pain, or complicate subsequent operations. Laparoscopy produces less peritoneal trauma than does conventional laparotomy and may result in decreased adhesion formation. We present a review of the available data on laparoscopy and adhesion formation, as well as laparoscopic adhesiolysis. We also review current adjuvant techniques that may be used by practicing laparoscopists to prevent adhesion formation.

[Specific features of medico-diagnostic strategy for cholesterosis of the gallbladder].

The work shows the incidence, criteria of the diagnosis and indications for surgical treatment of patients with cholesterosis of the gallbladder. An analysis of the results of examination and treatment of 3426 patients with diseases of the gallbladder has shown that the ultrasonic method of diagnostics allows detection of the cholesterosis form and determination of treatment strategy.

[Is a unified concept of species in ornithology possible? (An opinion of a practicing taxonomist)]. / Vozmozhna li edinaia kontseptsiia vida v ornitologii? (Mnenie praktikuiushchego sistematika).

An analysis of current publications on species problem reveals the radical discrepancy between the statements of species reality in nature and different number of these "realities" in light of different species concepts. It has been shown, that the only realities are populations, and "species" is only logical construct--the notion. A concept of species is a system of interconnected notions, where the main element--ontology of species has been always given a priori to the real biodiversity. For that reason every concept of species creates its own "virtual reality". Various concepts of species are not mutually comparable, and can not be criticized from each other's views and from position of "reality". Biological and Phylogenetic Species Concepts competing in ornithology exclude each other completely, and the unified concept of species is principally impossible. But as generally accepted species "standard" is needed, the unified convention of species must take the place of the various concepts of species.

[A commercial immunoenzyme reagent for the determination of ceruloplasmin]. / Kommercheskii immunofermentnyi diagnostikum dlia opredeleniia tseruloplazmina.

A diagnostic agent for enzyme immunoassay of ceruloplasmin has been developed. The method is highly sensitive and specific. The minimal detectable amount of ceruloplasmin is 25 ng/ml, variance coefficients with 1 lot being 2.6-7.3 percent and 4.2-10.8 percent within 3 lots. Blood serum ceruloplasmin concentration in health has made up 0.9 to 1.3 mg/ml. Ceruloplasmin levels were regularly shifted in Wilson's disease, cerebral lymphomas, sepsis, injuries, liver cirrhosis and other diseases.

[Evaluation of the clinical significance of determining the level of plasma fibronectin in acute poisoning]. / Otsenka klinicheskoi znachimosti opredeleniia urovnia plazmennogo fibronektina pri ostrykh toksicheskikh porazheniiakh.

Changes in plasma fibronectin (FN) levels in acute toxic conditions have been studied in 24 patients. A significant reduction of FN concentration was found within the first days of the disease; further changes in the FN levels are related to the disease outcome and have a prognostic value. An inverse correlation was found between the FN concentration and other blood biochemical parameters (LDH, creatinine, and urea). Developed renal insufficiency in the presence of hypofibronectinemia suggests a poor prognosis of the disease.

[Plasma ceruloplasmin in patients with hepatorenal insufficiency]. / Plazmennyi tseruloplazmin u bol'nykh s pechenochno-pochechnoi nedostatochnost'iu.

Changes in plasma ceruloplasmin levels were studied by enzyme immunoassay in 81 patients with the acute hepatorenal insufficiency syndrome (AHIS). Changed plasma ceruloplasmin concentration was detected in the first days of the disease, this change depending on AHIS severity and unrelated to the disease cause. A lowering of plasma ceruloplasmin level below 0.75 mg/ml is considered to be a poor prognostic sign. Measurement of plasma ceruloplasmin is a test available for clinical laboratories, it is highly sensitive, accurate, and reproducible.