General dynamics of the URT microbiome and microbial signs of recovery in COVID-19 patients.

COVID-19 is caused by an airborne virus, SARS-CoV-2. The upper respiratory tract (URT) is, therefore, the first system to endure the attack. Inhabited by an assemblage of microbial communities, a healthy URT wards off the invasion. However, once invaded, it becomes destabilised, which could be crucial to the establishment and progression of the infection. We examined 696 URT samples collected from 285 COVID-19 patients at three time-points throughout their hospital stay and 100 URT samples from 100 healthy controls. We used 16S ribosomal RNA sequencing to evaluate the abundance of various bacterial taxa, α-diversity, and ß-diversity of the URT microbiome. Ordinary least squares regression was used to establish associations between the variables, with age, sex, and antibiotics as covariates. The URT microbiome in the COVID-19 patients was distinctively different from that of healthy controls. In COVID-19 patients, the abundance of 16 genera was significantly reduced. A total of 47 genera were specific to patients, whereas only 2 were unique to controls. The URT samples collected at admission differed more from the control than from the samples collected at later stages of treatment. The following four genera originally depleted in the patients grew significantly by the end of treatment: Fusobacterium, Haemophilus, Neisseria, and Stenotrophomonas. Our findings strongly suggest that SARS-CoV-2 caused significant changes in the URT microbiome, including the emergence of numerous atypical taxa. These findings may indicate increased instability of the URT microbiome in COVID-19 patients. In the course of the treatment, the microbial composition of the URT of COVID-19 patients tended toward that of controls. These microbial changes may be interpreted as markers of recovery.

[Metabolic characteristics, nutritional status and behavior factors in people with obesity, working at industrial plant].

An obesity is the topical problem worldwide, because excessive weight is the reason of metabolic changes that lead to development of serious chronic non-communicable diseases. The examination was aimed to assess clinical and metabolic health, nutritional status and behavior risk factors of obesity in people, working at the industrial plant. Material and methods. In 348 workers of the industrial plant (210 males, 138 females) examination was performed including anthropometry: body mass index (BMI), waist and hip circumferences and their ratio, blood pressure level detection, laboratory blood tests (glucose and cholesterol levels), and questionnaire on nutritional habits and physical activity. All examined were divided in three groups depending on BMI: 1st group - 151 (43.3%) persons with normal body mass, 2nd group - 125 (36%) working people with overweight and 3rd group - 72 (20.7%) people with obesity. Results. More than half of examined workers (mostly females) had overweight or obesity with predominantly abdominal type of fat distribution and I degree. Arterial hypertension was diagnosed in 41% of all working people (predominantly I degree), more frequently observed in obese (62.5%) and in males (45%). Each fourth man and woman had elevated total cholesterol level, which increased depending on BMI raise. Postprandial hyperglycemia was observed in 5.5% of cases, with the same frequency in both genders, predominantly in obesity. A significant part of responded with obesity, especially females, confirmed, that the reasons of excessive weight were stresses and somatic diseases (37%), and they preferred chocolate from confectionery. It was noted a correlation of BMI with egg consumption in males and with the consumption frequency of nuts, seeds and chocolate in females. Physical activity less than 3 times a month or never was observed in most of responded workers with overweight and obesity (73%), especially in males, as well as in females during the last month before the survey (57%). Conclusion. Preventive examination detected not only high prevalence of abdominal obesity, arterial hypertension and fat metabolism disturbances in people, working at the industrial plant, but also allowed to detect the most significant alimentary and behavioral risk factors of obesity.

Catching a virus in a molecular net.

A metal-organic molecular net composed of tannic acid (TA) and iron(iii) was constructed around the brome mosaic virus (BMV) particle to determine whether the added net could act as a transport barrier for water, and if the net could stabilize the virus in physically or chemically challenging environments. This new virus engineering strategy is expected to provide benefits both in the study and technological applications of viruses. For instance, a virus wrapped in a thin molecular layer could be extracted from solution either in air or vacuum, and its structure, composition and even internal dynamics could be interrogated by methods not compatible with a liquid environment. Atomic force microscopy (AFM) studies of Fe(iii)-TA coated BMV in liquid and in air supported a marked resistance to dehydration when compared to wtBMV. Native charge detection mass spectrometry (CDMS), was employed to estimate the number of molecules in the molecular net which wrapped the virus. The CDMS data suggested that less than one molecular monolayer wrapped the virus. Additionally, it was found, that this very thin molecular coat was sufficient to render the coated viruses resistant to storage conditions that typically lead to virus disassembly over time. A temporary coat imparting increased resistance to disassembly could be useful in adding time delay control or alleviate required storage conditions of engineered viruses for therapeutic purposes.

Nanoindentation of Isometric Viruses on Deterministically Corrugated Substrates.

It has been just over 100 years since inventor Joseph Coyle perfected the egg carton-a package format that has known very little changes since its first appearance ( Dhillon , S. B. C. Inventor Created Better Way to Carry Eggs. In The Globe and Mail Vancouver , 2013 ). In this article, we extend Coyle's old idea to the study of mechanical properties of viruses. Virus stiffness, strength, and breaking force obtained by force spectroscopy atomic force microscopy (AFM) provide the knowledge required for designing nanocontainers for applications in biotechnology and medicine, and for understanding the fundamentals of virus-host interaction such as virus translocation from one cellular compartment to another. In previous studies, virus particles adsorbed on flat surfaces from a physiological buffer were subjected to directional deformation by a known force exerted via a microscopic probe. The affinity between the virus shell and surface is required to be strong enough to anchor particles on the substrate while they are indented or imaged, yet sufficiently weak to preserve the native structure and interactions prior deformation. The specific question addressed here is whether an experimental scheme characterized by increased contact area and stable mechanical equilibrium under directional compression would provide a more reliable characterization than the traditional flat substrate approach.

Molecular Mechanisms of Ceftaroline Susceptibility Reduction in Methicillin-Resistant Staphylococcus aureus.

Ceftaroline is a unique cephalosporin with activity against methicillin resistant Staphylococcus aureus (MRSA). It was approved for clinical use in the USA, Europe and Russian Federation since 2010 for the treatment of the skin and soft tissue infection and community-acquired pneumoniae. In the present study there was used molecular typing of 24 isolates of MRSA with reduced susceptibility to ceftaroline. For 8 isolates belonging to different genetic lines (ST8, ST239 and ST228) and requiring MICs there were determined antibiotic concentrations preventing formation of resistant mutants (mutant prevention concentration) and the ranges of the mutant selection window (MSW). The last majority of the isolates with reduced susceptibility to ceftaroline (MIC of 2 mcg/ml) belonged to the clonal line ST228. The whole genome sequencing of two isolates of ST228 showed that they belonged to the epidemic South Germany genetic line and were characterized by the presence of mutations in PBP2a (N146K) and PBP2 (C197Y) responsible for reduced susceptibility. The highest rates of MPC (32 mcg/ml) and MSW (2-16 mcg/ml) were observed in the clinical isolates belonging to the genetic line ST8. The isolates of ST239 and ST228 had the selection window within 2-4 mcg/ml. No dependence of the MIC and MPC/MSW levels was detected.

[Clinico-morphological substantiation of feasibility of angiosomic adipose-cutaneous flaps from temporal and parietal regions application].

Actuality of the problem, consisting of restoration of defects and deformity zones on head and neck, is caused by great quantity of patients, surgical treatment of which constitute important medico-social problem. The flaps, raised from temporal and parietal regions, are considered perspective for replacement of defects and the deformity zones on head and neck. But, for their correct isolation and mobilization it is necessary to know the topographoanatomical peculiarities of superficial temporal artery, localization of which differs, depending on the head form. The methods of isolation and mobilization of angiosomic preauricular, postauricular and fascial temporal-parietal flaps were optimized in the clinic, basing on analysis of results of the investigations performed and computeric modeling of the superficial temporal artery branches.

[Methods for determining the activity of lysosomal hydrolases used in diagnosis of hereditary lysosomal diseases].

The paper describes biochemical methods for determining the activity of a number of lysosomal hydrolases, deficiency of which underlies hereditary lysosomal storage diseases. The methods permit the precise diagnosis of the diseases in a patient and the prenatal diagnosis of a disease in a fetus in at-risk families.

[Expression of gelatinases A and B and their endogenous regulators in immortal and transformed fibroblasts].

Matrix metalloproteinases (MMP) play a critical role in tumor invasion and metastasis. The aim of this study was to elucidate peculiarity of expression of gelatinases A and B (MMP-2 and MMP-9), membrane type MMP (MT1-MMP) and tissue inhibitor of MMP (TIMP-2) in immortal (IF) and transformed fibroblasts (TF).The study was carried out using embryo rat fibroblasts, sequentially immortalized with the polyomavirus LT gene and transformed with the E7 gene of human papilloma virus (HPV-16). Papilloma virus type 16 and 18 are etiological factors of cervical cancer. The primary fibroblast (PF) culture of Fisher rats was used as control. Analysis of TF and IF involved: determination of MMP-2 and MMP-9 activity by hydrolysis of specific substrate--radioactive collagen type IV; obtaining of MMP spectra by zimographic assay and estimation of the mRNA expression (by RT-PCR) of MMP-2, MMP-9, MT1- MMP and TIMP-2. It was found: 1) collagenolytic activity of MMP was increased only in TF and was dependent on the degree of cell tumorogenity; 2) the study of MMP spectra was shown that MMP-9 was found in TF only but MMP-2 was found in all investigated clones; 3) The mRNA expression of MMP-9, MT1-MMP and TIMP-2 was increased in all TF while the MMP-2 expression was increased in TF only after TF cell selection on rats; 4) The collagenolytic activity as well as the mRNA expression of MMP-2 and MMP-9 themselves and of MMP-2 endogenous regulators (MT1-MMP and TIMP-2) did not change in immortalized fibroblasts compared to PF. The data obtained indicate changes in the enzyme/inhibitor/activator ratio and also suggest of a significant increase in the TF destructive potential. MMP-9 is supposed to be a marker of fibroblasts transformed by E7 HPV-16 gene in cell culture.

[A surgical treatment for neovascular terminal glaucoma (preliminary communication)].

In 40 patients with terminal neovascular glaucoma of various genesis (visual acuity 0 to wrong light projection), 4 rectus muscles were tightly ligated at the insertion site of their strings to the sclera of the eyeball with silk threads under local anesthesia. The threads were not postoperatively removed and left under the conjunctiva (Invention Patent No. 2286752 of November 10, 2006). The proposed procedure for blocking the anterior ciliary arteries may be used as the method of choice in patients with terminal neovascular glaucoma. It makes it possible to achieve a steady-state reduction in intraocular pressure, by improving humor outflow from the corner of the anterior chamber, to diminish or completely eliminate pain syndrome and corneal edema, by maintaining the mobility of the eye in corpore, and to avoid enucleation.

[Distribution of mutations of acid beta-D-glucosidase gene (GBA) among 68 Russian patients with Gaucher's disease].

Gaucher disease (GD) is the most frequent lysosomal storage disease presenting in all populations. Mutations in the acid beta-D-glucosidase gene (GBA) cause development of GD, resulting in a decrease or full loss of activity of this enzyme. We report here the results of the molecular-genetic analysis in 68 Russian GD patients from 65 families with the three types of the disease. We have identified 126 mutation alleles from 136 investigate alleles. In addition to known mutations p.N370S, c.1263-1317del (del55), p.L444P, p.R463C, Rec NciI, we identified rare mutations p.R120W, p.R170C, p.W184R, p.G202R, Rec C, presenting in other populations and mutations p.P236T, p.L249Q, p.L288P, p.P319S, p.V352M, p.W381X, p.A384D which are had not been described before.

[Some factors influencing the efficiency of disk-diffusion test to determine the antibiotic sensitivity of microorganisms].

On diffusion of an antibiotic from the disk to the nutrient medium, gel concentration of an antibiotic changes. This process is individual for each antibiotic. The action of different concentrations of the antibiotic at different intervals results in the suppression of microbial growth around the disk (formation of a zone). The relationship between the antibiotic concentration in the disk and the diameter of a microbial growth suppression area is proportional in the limited range of concentrations. The relationship between the concentration of an antibiotic and the diameter of a growth suppression area may greatly vary for various taxonomic groups of microorganisms and for various strains. The established fact may be responsible for the disagreement of results of a microbial antibiotic sensitivity test, by using different methods and makes the standardization of the disk-diffusion test difficult.

Study on the substituents' effects of a series of synthetic chalcones against the yeast Candida albicans.

A large series of chalcones were synthesized and studied for activity against Candida albicans. The SAR analysis showed that the antifungal activity was highly dependent on the substitution pattern of the aryl rings and correlated to a large extent with the ability of compounds to interact with sulfhydryl groups. The most active were the hydroxylated chalcones as their activity related to the location of the phenolic group in the aryl ring B as follows: o-OH>p-OH approximately 3,4-di-OH>m-OH. These and other correlations obtained strongly contribute to the knowledge for design of anticandidal chalcones.

[Lysosomal cysteine proteinases at neoplastic transformation].

This review summarizes the data on classification, properties, structure and biological functions of mammalian papain-like lysosomal cysteine proteinases (cathepsins). The most studied are cathepsins B, H, L, S and K. The peculiarities of lysosomal cysteine proteinase regulation, particularly regulation by specific endogenous inhibitors are discussed. Involvement of these enzymes in development of physiological and pathological processes are considered. Special attention is paid to the role of cysteine proteinases and their inhibitors in neoplastic transformation. The possibility of use cathepsin endogenous inhibitors as basis for design antitumorogenic therapeutic substances are discussed.

Antimicrobial and cytotoxic activity of Ruta graveolens.

The methanol, petroleum ether, ethyl acetate and water-methanol extracts of Ruta graveolens were found to possess antimicrobial and cytotoxic activities.

[Tripeptidyl peptidase 1 deficiency in neuronal ceroid lipofuscinosis. A novel mutation]. / Nedostatochnost' tripeptidil peptidazy 1 pri neironal'nom tseroidnom lipofuktsinoze. Novaia mutatsiia.

The data on biochemical and molecular-genetic diagnostics of a hereditary lisosomal storage disease, late infantile neuronal ceroid lipofuscinosis (CLN2) are presented. The disease is associated with a hereditary deficiency of pepstatin-unsensitive peptidase--tripeptidylpeptidase 1 (TPP1)--caused by mutations in the TPP1-coding gene CLN2. Among the 30 patients with clinical manifestations of CLN, six patients with a pronounced decrease in TPP1 activity were revealed; these data were interpreted as indicating the presence of CLN2 in these patients. The analysis of the isolated DNA indicated the availability of the most widespread mutation g3670 C > T(R208X) leading to the untimely termination of TPP1 synthesis. It was shown that in 5 patients this mutation is present in homozygous state and in one patient, in the heterozygous state. In this patient a hitherto unknown mutation, g3665G > A (R206H), was revealed. The pathogenetic significance of this mutation and the importance of molecular-genetic diagnosis of CLN are discussed with regard to medico-genetic consulting and prenatal diagnosis of this disease.

New bioactive chalcones in propolis from El Salvador.

2',3'-Dihydroxy-4,4'-dimethoxychalcone (1) and 2',3',4-trihydroxy-4'-methoxy-chalcone, two new chalcones, were isolated from propolis from El Salvador. The compounds showed significant antibacterial and antifungal activity and moderate toxicity to Artemia salina nauplii.

The first glycosides isolated from propolis: diterpene rhamnosides.

Two diterpene glycosides, ent-8(17)-labden-15-O-alpha-L-rhamnoside and ent-8(17)-labden-15-O-(3'-O-acetyl)-alpha-L-rhamnoside (new natural compounds) were isolated from propolis from El Salvador. The compounds showed significant antibacterial activity and moderate toxicity to Artemia salina nauplii. These are the first glycosides reported in bee glue.