Affimer proteins for F-actin: novel affinity reagents that label F-actin in live and fixed cells

Imaging the actin cytoskeleton in cells uses a wide range of approaches. Typically, a fluorescent derivative of the small cyclic peptide phalloidin is used to image F-actin in fixed cells. Lifeact and F-tractin are popular for imaging the cytoskeleton in live cells. Here we characterised novel affinity reagents called Affimers that specifically bind to F-actin in vitro to determine if they are suitable alternatives as eGFP-fusion proteins, to label actin in live cells, or for labeling F-actin in fixed cells. In vitro experiments showed that 3 out of the 4 Affimers (Affimers 6, 14 and 24) tested bind tightly to purified F-actin, and appear to have overlapping binding sites. As eGFP-fusion proteins, the same 3 Affimers label F-actin in live cells. FRAP experiments suggest that eGFP-Affimer 6 behaves most similarly to F-tractin and Lifeact. However, it does not colocalize with mCherry-actin in dynamic ruffles, and may preferentially bind stable actin filaments. All 4 Affimers label F-actin in methanol fixed cells, while only Affimer 14 labels F-actin after paraformaldehyde fixation. eGFP-Affimer 6 has potential for use in selectively imaging the stable actin cytoskeleton in live cells, while all 4 Affimers are strong alternatives to phalloidin for labelling F-actin in fixed cells

The Relationship of Urinary Metabolites of Carbaryl/Naphthalene and Chlorpyrifos with Human Semen Quality

Most of the general population is exposed to carbaryl and other contemporary-use insecticides at low levels. Studies of laboratory animals, in addition to limited human data, show an association between carbaryl exposure and decreased semen quality. In the present study we explored whether environmental exposures to 1-naphthol (1N), a metabolite of carbaryl and naphthalene, and 3,5,6-trichloro-2-pyridinol (TCPY), a metabolite of chlorpyrifos and chlorpyrifos-methyl, are associated with decreased semen quality in humans. Subjects (n = 272) were recruited through a Massachusetts infertility clinic. Individual exposures were measured as spot urinary concentrations of 1N and TCPY adjusted using specific gravity. Semen quality was assessed as sperm concentration, percent motile sperm, and percent sperm with normal morphology, along with sperm motion parameters (straight-line velocity, curvilinear velocity, and linearity). Median TCPY and 1N concentrations were 3.22 and 3.19 μg/L, respectively. For increasing 1N tertiles, adjusted odds ratios (ORs) were significantly elevated for below-reference sperm concentration (OR for low, medium, and high tertiles = 1.0, 4.2, 4.2, respectively; p-value for trend = 0.01) and percent motile sperm (1.0, 2.5, 2.4; p-value for trend = 0.01). The sperm motion parameter most strongly associated with 1N was straight-line velocity. There were suggestive, borderline-significant associations for TCPY with sperm concentration and motility, whereas sperm morphology was weakly and nonsignificantly associated with both TCPY and 1N. The observed associations between altered semen quality and 1N are consistent with previous studies of carbaryl exposure, although suggestive associations with TCPY are difficult to interpret because human and animal data are currently limited

Relationship of cytokine level dynamics with longterm results of early rheumatoid arthritis therapy

The aim of the work – to research the relationship between the dynamics of cytokine levels in the early period (the first 12 and 24 weeks) of pharmacotherapy for rheumatoid arthritis (RA) and long-term outcomes in patients with the disease.Material and methods. The analysis included 93 patients with early RA. The majority of patients were women (n=77), middle-aged (58 [49; 66] years), with an early stage of the disease (the duration of the disease is 7 [4–11.5] months), seropositive according to IgM rheumatoid factor and cyclic citrullinated peptide antibodies, who had high (59.7%) or moderate (38.8%) disease activity.The concentration of 27 cytokines in the blood serum was determined with the help of multiplex xMAP technology on the Bio-Plex array system analyzer (BIORAD, USA). Repeated clinical examination of patients was carried out after 6 years.Results. Patients who achieved remission/low disease activity SDAI after 6 years had a lower level of IL-6 (7.7 [7.4; 23.3]), IL-9 (13.5 [9.1; 18.9]) 12 weeks after the start of therapy; lower IL-9 level (12.6 [6.8; 16.2]) 24 weeks after the start of the treatment, compared with the group of patients with moderate and high inflammatory activity (23.5 [12.4; 69.5], 17.8 [15; 29] and 18.5 [14.2; 22.8] respectively).Findings. The assessment of the level of proinflammatory cytokines (IL-6, IL-17), immunoregulatory cytokine IL-9 allows to evaluate the activity of the disease more fully and identify a group of patients, who needs the therapy intensification

IСP-MS ANALYSIS OF HONEY AND BEE PRODUCTS

The microelement composition and isotopic ratios (87Sr/86Sr) of bee tissues and apicultural products from the Perm Krai, Republic of Bashkortostan, and Krasnodar Krai have been investigated. The obtained statistical data allowed the meads to be classified

Practical guidelines for standardising the measurement of resting metabolism by indirect calorimetry: a literature review

Accurate resting metabolic rate readings are essential for dietary planning and body composition monitoring not only for healthy individuals but also for athletes. A number of factors can alter resting metabolic rate during its measurement by indirect calorimetry. The methodology used may affect the results of the study. A clear standardisation of this procedure is needed to obtain the most accurate results.Purpose: To review the literature to determine the optimal subject condition and methodology for the resting metabolism measurement procedure using indirect calorimetry.Materials and methods: A literature search was conducted in PubMed, MEDLINE and Cochrane Library databases. The query included key words and logical phrases: “calorimetry”, “indirect calorimetry”, “resting metabolic rate”, “energy metabolism”, “basal metabolism”, “standards”. Only Englishlanguage studies and human studies were considered. Additional information was identified because of the review and included in the review.Results: the parameters of standardization during the resting metabolism measurement procedure are described: consumption of food, ethanol, caffeine, nicotine; daily activities and physical activity; body position in space and environmental conditions during the measurement; actions of the specialist performing the procedure, etc. The article outlines effective methods for measuring resting metabolism to obtain the most accurate results in both athletes and non-athletes.Conclusion: an attempt has been made to formulate precise methodological rules for standardization and recommendations for measuring resting metabolism by indirect calorimetry

Clinical features of ACPA-negative and ACPA-positive variants of rheumatoid arthritis

The aim of the study was to study the features of the clinical picture of the disease in patients with ACPA-negative and ACPA-positive variants of rheumatoid arthritis.Materials and methods. The study included patients with a reliable diagnosis of rheumatoid arthritis (RA) according to the criteria of ACR/EULAR 2010. Depending on the values of the ACPA, two groups of patients were recruited: ACPA-positive and ACPA-negative, comparable in gender, age, duration of the disease and therapy. The nature of the onset and course of the disease, the activity of RA were evaluated (according to the DAS28, SDAI, CDAI indices).Results and discussion. The study included 79 patients with ACPA-negative variant of RA and 79 with ACPA-positive. Age of patients (Me [IR], in years) with the ACPA(–) variant was 52 [39; 62], with the ACPA(+) – 54 [42; 62], the duration of the disease (in months) is 59 [23; 122] and 48 [17; 84] respectively. In ACPA(+) patients, higher disease activity was determined by the indices DAS28-CRP, DAS28-ESR, SDAI, CDAI, values of C-reactive protein and erythrocyte sedimentation rate, a greater number of painful and swollen joints (p<0.05). According to the localization of the involved joints, arthritis of the proximal interphalangeal, metacarpal, wrist and shoulder joints was more often determined in ACPA(+) patients. Systemic manifestations of RA at the time of examination and in the anamnesis were statistically significantly more common in ACPA(+) (32.9%) than in ACPA(–) (17.7%). Of the systemic manifestations, rheumatoid nodules were more common in ACPA(+) patients, a tendency to a higher frequency of neuropathy, scleritis and episcleritis was revealed in ACPA(–) patients.Conclusion. In patients with ACPA(–) subtype, clinical signs of joint damage and the inflammatory component are less pronounced compared to ACPA(+). However, the mixed picture of manifestation, the less “bright” course of the disease, the absence of characteristic immunological biomarkers necessitate long-term and careful monitoring of this group of patients. At the same time, the subjective severity of the disease and dysfunction due to ankylosing joints do not differ from the ACPA(+) variant of RA

Practical recommendations for safe reduction of body weight in combat sports: scoping review

The presence of weight categories in martial arts requires an athlete to constantly maintain and control body weight and, in some cases, to reduce it. For this reason, it is common among martial artists to use forced methods of body weight reduction, which are often unphysiological and may have a negative impact on performance, as well as pose a danger to health. In this connection, it is of practical interest to search, analyse and introduce into practice physiological and safe methods of body weight reduction by athletes-athletes before participation in competitions. The aim of the study is to evaluate the safety and effectiveness of various methods of weight loss in athletes preparing to participate in competitions.Materials and methods: the review was conducted according to the methodology of Scoping review. Articles were selected in the domestic data-bases eLibrary and Russian State Library, using the following keywords: «методы снижения массы тела ИЛИ снижение массы тела ИЛИ весогонка» И «единоборства ИЛИ боевые виды спорта», as well as in the foreign databases ScienceDirect and PubMed, using the following keywords: «making weight OR weight loss» AND «combat sports», placed in a search depth of 15 years.Results: we found 121 studies, 3 duplicates were excluded after initial screening, 118 studies were screened for inclusion criteria, and a total of 16 studies were included in the review.Conclusion: a reduction of 5 to 10 % of body weight less than 7 days before the official weigh-in may adversely affect performance parameters and the state of the body’s functional systems. A reduction of up to 5 % of body weight 7 or more days before the official weigh-in is the safest

Improved method for the obtaining DTTA-appended 2,2’-bipyridine ligands for lanthanide cations

The composition of the reaction mixture after DTTA tert-butyl ester alkylation with 6'-halomethyl-5-phenyl-2,2'-bipyridines was studied. In addition to the target product, DTTA-appended 2,2’-bipyridine, the corresponding 6'-hydroxymethyl-substituted 2,2’-bipyridine and (5'-phenyl-[2,2'-bipyridin]-6-yl)methyl formate were isolated as by-products in some cases. Finally, an improved procedure for the DTTA tert-butyl ester alkylation with 6'-halomethyl-5-phenyl-2,2'-bipyridines by using Finkelstein reaction was developed

Practical guidelines for standardising the measurement of resting metabolism by indirect calorimetry: a literature review

Accurate resting metabolic rate readings are essential for dietary planning and body composition monitoring not only for healthy individuals but also for athletes. A number of factors can alter resting metabolic rate during its measurement by indirect calorimetry. The methodology used may affect the results of the study. A clear standardisation of this procedure is needed to obtain the most accurate results.Purpose: To review the literature to determine the optimal subject condition and methodology for the resting metabolism measurement procedure using indirect calorimetry.Materials and methods: A literature search was conducted in PubMed, MEDLINE and Cochrane Library databases. The query included key words and logical phrases: “calorimetry”, “indirect calorimetry”, “resting metabolic rate”, “energy metabolism”, “basal metabolism”, “standards”. Only English-language studies and human studies were considered. Additional information was identified because of the review and included in the review.Results: the parameters of standardization during the resting metabolism measurement procedure are described: consumption of food, ethanol, caffeine, nicotine; daily activities and physical activity; body position in space and environmental conditions during the measurement; actions of the specialist performing the procedure, etc. The article outlines effective methods for measuring resting metabolism to obtain the most accurate results in both healthy individuals and athletes.Conclusion: an attempt has been made to formulate precise methodological rules for standardisation and recommendations for measuring resting metabolism by indirect calorimetry

Pathological anatomy of the new coronavirus infection COVID-19 according to autopsy materials 2020

Introduction. Organ lesions due to the new coronavirus infection are determined by several factors: direct cytopathic effect of SARS-CoV-2, release of an unbalanced amounts of proinflammatory cytokines leads to systemic damage of the vascular endothelium and increasing hypoxia. These factors in total result in organ dysfunction.Methods and materials. We examined 96 pathological studies’ protocols of patients who died from the PCR-confirmed new coronavirus infection. Histological slides stained with hematoxylin and eosin were also analyzed.Results. Macroscopical and microscopical changes seen mostly in lungs, myocardium and other internal organs were described. We reviewed main causes of death (pulmonary and cardiopulmonary failure, multiple organ dysfunction syndrome) and most common concomitant diseases (diabetes mellitus, essential hypertension, obesity). The morphological features of diffuse alveolar damage in the acute and proliferative phases were described in detail. Taken together with the synchronous involvement of the microvasculature in the pathological process it became the main cause of death. The most severe structural changes in lungs were seen after 20 days of disease. These changes were correlated with dysregenerative metaplastic and dysplastic processes. Processes of disturbed regeneration reflect both the severe viral transformation of the alveolar epithelium and the fact that regeneration developed under mechanical ventilation