[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.

[Treatment of chronic allergic blepharoconjunctivitis].

OBJECTIVE: to evaluate the effectiveness of therapeutic eyelid hygiene in the treatment of chronic, or perennial, allergic blepharoconjunctivitis (ABC). MATERIAL AND METHODS: A total of 30 patients with chronic ABC (11 men, 19 women aged 25-41 years; disease duration 2-4 years) were examined before and after a treatment course which included instillations of olopatodine hydrochloride instillations (1 mg/ml, 2 times daily) and hyaluronic acid (2 mg/ml, 3-4 times daily). Patients from the main group (18 patients, 36 eyes) additionally practiced therapeutic eyelid hygiene using Blepharoshampoo, Blepharolotion or Blepharosalfetka, and Blepharogel-1 (2 times daily). The control group was given only anti-allergic therapy and tear substitutes. Conventional ophthalmological examination, allergy testing, ABC signs and symptoms evaluation, Schirmer's 1 and Norn's tests, tear film break-up time evaluation, assessment of meibomian gland function, optical coherence tomography (OCT) for tear meniscus, xerosis index evaluation and lissamine green staining for lid wiper epitheliopathy, and anterior segment photography with further computer morphometry were performed in all patients. RESULTS: Dry eye signs and symptoms, including combined lipid, water, and mucus tear deficiency, were found in all patients with chronic ABC. Patients from the main group demonstrated a significantly greater decrease of integral indices of subjective ocular discomfort, reduction of chronic ABC signs as well as more pronounced improvement of the meibomian gland function, appearance of the lid wiper region, tear film break-up time, results of the OCT-meniscometry, and xerosis index. CONCLUSION: Introduction of therapeutic eyelid hygiene to the complex treatment of chronic allergic blepharoconjunctivitis with the aim of resolving secondary dry eye symptoms enhances the effectiveness of the treatment.

[Mechanism of population density compensation in island bumblebee taxocenoses (Hymenoptera, Apidae, Bombus) and the notion of reserve compensatory species].

The notion of a dynamic compensatory system is discussed, characterized by the alternation of species occupying the leading position in bumblebee taxocenoses, while the total density of these pollinators in island ecosystems remains at similar levels. The functioning of the compensatory system is regulated by both abiotic factors (the weather and climate) and biotic factors (competition for trophic resources). The stability of the system is determined by the presence of reserve compensatory species capable of rapid population growth against the background of depressed abundance of other species under changing environmental conditions.

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.

A transgenic cell line with inducible transcription for studying (CGG)n repeat expansion mechanisms.

There are more than 30 inherited human disorders connected with repeat expansion (myotonic dystrophy type I, Huntington's disease, Fragile X syndrome). Fragile X syndrome is the most common reason for inherited intellectual disability in the human population. The ways of the expansion development remain unclear. An important feature of expanded repeats is the ability to form stable alternative DNA secondary structures. There are hypotheses about the nature of repeat instability. It is proposed that these DNA secondary structures can block various stages of DNA metabolism processes, such as replication, repair and recombination and it is considered as the source of repeat instability. However, none of the hypotheses is fully confirmed or is the only valid one. Here, an experimental system for studying (CGG)n repeat expansion associated with transcription and TCR-NER is proposed. It is noteworthy that the aberrations of transcription are a poorly studied mechanism of (CGG)n instability. However, the proposed systems take into account the contribution of other processes of DNA metabolism and, therefore, the developed systems are universal and applicable for various studies. Transgenic cell lines carrying a repeat of normal or premutant length under the control of an inducible promoter were established and a method for repeat instability quantification was developed. One type of the cell lines contains an exogenous repeat integrated into the genome by the Sleeping Beauty transposon; in another cell line, the vector is maintained as an episome due to the SV40 origin of replication. These experimental systems can serve for finding the causes of instability and the development of therapeutic agents. In addition, a criterion was developed for the quantification of exogenous (CGG)n repeat instability in the transgenic cell lines' genome.

Derivation of iPS cell line (ICGi032-A) from a patient affected with fragile X syndrome.

Trinucleotide repeat expansion diseases such as fragile X syndrome are of great interest to study since the mechanism of its development is still unknown. IPS cell lines are some of the most convenient models for studying. The ICGi032-A iPS cell line was obtained from the peripheral blood mononuclear cells of the patient affected with fragile X syndrome. ICGi032-A iPS cell line have a normal karyotype, expression of pluripotency markers and can differentiate in vitro into the cells of three germ layers.

Establishment of an induced pluripotent stem cell line (ICGi026-A) from peripheral blood mononuclear cells of a patient with fragile X syndrome.

Expansion over 200 CGG repeats in FMR1 gene causes inherited intellectual disability or autism spectrum disorder named as fragile X syndrome. Despite the known cause fragile X syndrome pathogenesis has not been specified yet. The ICGi026-A iPSCs line was obtained by the reprogramming of the peripheral blood mononuclear cells from a 9-year-old boy with fragile X syndrome. The ICGi026-A iPSCs expressed pluripotency markers, had a normal male karyotype (46, XY) and had the capacity to in vivo differentiate into the cells of three germ layers.