Proteomic and transcriptomic analyses of Cutibacterium acnes biofilms and planktonic cultures in presence of epinephrine.

Transcriptomic and proteomic analysis were performed on 72 h biofilms of the acneic strain Cutibacterium acnes and planktonic cultures in the presence of epinephrine. Epinephrine predominantly downregulated genes associated with various transporter proteins. No correlation was found between proteomic and transcriptomic profiles. In control samples, the expression of 51 proteins differed between planktonic cultures and biofilms. Addition of 5 nM epinephrine reduced this number, and in the presence of 5 µM epinephrine, the difference in proteomic profiles between planktonic cultures and biofilms disappeared. According to the proteomic profiling, epinephrine itself was more effective in the case of C. acnes biofilms and potentially affected the tricarboxylic acid cycle (as well as alpha-ketoglutarate decarboxylase Kgd), biotin synthesis, cell division, and transport of different compounds in C. acnes cells. These findings are consistent with recent research on Micrococcus luteus, suggesting that the effects of epinephrine on actinobacteria may be universal.

Changes in the Transcription of Proliferation- and Apoptosis-Related Genes in Embryos in Women of Different Ages under the Influence of Extracellular Vesicles from Donor Follicular Fluid In Vitro.

We studied the influence of extracellular vesicles from the follicular fluid of a young donor on gene expression (MKI67, MYBL2, CCNB1, CCND1, CCNE1, CALM2, BAX, NDRG1, TP53I3, VEGF, VCAN, HAS2, CTSL2, PIBF1, RPL37, PFKP, GPX3, and AQP3) in embryos of women of different ages. According to nanoparticle tracking analysis data, the concentration of extracellular vesicles was 3.75±0.47×1011 particles/ml and the mean particle size was 138.78±9.90 nm. During co-culturing of the follicular fluid extracellular vesicles with blastocysts of young women, we observed significantly increased expression of mRNA for genes CTSL2, CCND1, CCNE1, VEGF and reduced expression of BAX gene mRNA in comparison with embryos in women of late reproductive age. We hypothesized that addition of extracellular vesicles of the oocyte follicular fluid from a young donor to the culture medium of embryos could slow down apoptosis process typical of blastocyst cells in women above 36 years.

Investigation of the variability of the pterygomandibular space from the point of view of the target point of the inferior alveolar nerve block.

INTRODUCTION: A thorough knowledge of the anatomy and variability of the pterygomandibular space (PMS) is essential to the success of local anesthesia. This study is aimed at determining the volume of the PMS depending on the shape of the skull, face and mandible using the computed tomography (CT). MATERIALS AND METHODS: anonymized computed tomograms of 48 adult patients were analyzed, the indicators of the cranial index (CI), facial index (FI) according to Yzard, the high lengthy index of the mandible (HLI), the latitudinal-altitude index of the mandible (LAI), the longitudinal latitude index of the mandible (LLI) were calculated. RESULTS: Differences in the volume of the PMS were found depending on the shape of the skull, face and mandible. When determining the CI, the largest volume of space on the right was observed with brachycrania (2.05 ml), on the left - with mesocrania and brachycrania (2.0 ml each). With regard to the FI, the largest volume of space was obtained with medium face on the right side (2.03 ml) and broad and medium types of faces (2.0 ml each) on the left. When calculating the indicators for the shapes of mandible, the largest volume of space was found in the brachygenic (2.05 ml), leptogenic (1.98 ml) and platigenic (1.97 ml) shapes on the right and leptogenic and platigenic shapes on the left (2.0 ml each). There were statistically significant differences in the volume of the right side of the PMS between different shapes according to the cranial index (F = 5.075; p = 0.0095). The most pronounced difference was present between samples with brachycrania and dolichocrania: 0.35 mm³ (95% CI 0.05-0.65); p = 0.0188. There was a statistically significant correlation with sex for parameters on both the right and left sides. The values of indicators in men exceeded those of measurements obtained from women. CONCLUSION: The data obtained determine the differences in the volume of the PMS in individuals with different shapes of the skull, face and mandible, as well as sex differences. Thus, indicating the need to take into account these features when choosing the required amount of anesthetic in clinical practice. The average volume of the PMS space, according to our data, is 1.8-2.0 ml, which is consistent with the clinical data described in the literature.

Female and Male Phenotypes of Iron Deficiency in CHF. Additional analysis of the «The Prevalence of Iron Deficiency in Patients With Chronic Heart Failure in the Russian Federation (J-CHF-RF)¼ study.

Aim    To evaluate the incidence of iron deficiency (ID) in men and women with chronic heart failure (CHF) and to compare clinical and functional indexes in patient with and without ID depending on the gender.Material and methods    An additional analysis of the study "Prevalence of Iron Deficiency in Patients With Chronic Heart Failure in the Russian Federation (ID-CHF-RF)" was performed. The study included 498 (198 women, 300 men) patients with CHF, in whom, in addition to iron metabolism, the quality of life and exercise tolerance (ET) were studied. 97 % of patients were enrolled during their stay in a hospital. ID was defined in consistency with the European Society of Cardiology (ESC) Guidelines. Also, and additional analysis was performed according to ID criteria validated by the morphological picture of the bone marrow.Results    ID was detected in 174 (87.9 %) women and 239 (79.8 %) men (p=0.028) according to the ESC criteria, and in 154 (77.8 %) women and 217 (72.3 %) men (p=0.208) according to the criteria validated by the morphological picture of the bone marrow. Men with ID were older and had more severe CHF. They more frequently had HF functional class (FC) III and IV (63.4 % vs. 43.3 % in men without ID); higher concentrations of N-terminal pro-brain natriuretic peptide (NT-proBNP) and lower ET. HF FC III increased the probability of ID presence 3.4 times (p=0.02) and the probability of HF FC IV 13.7 times (p=0.003). This clinical picture was characteristic of men when either method of determining ID was used. In women, ID was not associated with more severe CHF.Conclusion    Based on the presented analysis, it is possible to characterize the male and female ID phenotypes. The male ID phenotype is associated with more severe CHF, low ET, and poor quality of life. In females of the study cohort, ID was not associated with either the severity of CHF or with ET.

[CIC-rearranged sarcoma: a case report and literature review]. / CIC-perestroennaya sarkoma: klinicheskoe nablyudenie i obzor literatury.

CIC-rearranged sarcoma is a rare and extremely aggressive tumor that occurs mainly in soft tissues. Despite the fact that identification of a characteristic genetic rearrangement is necessary to verify the diagnosis, in most cases, the correct diagnosis can be made by comparing histological signs and a characteristic immunophenotype, which greatly speeds up the diagnosis. The article describes a case of CIC-rearranged sarcoma in a 14-year girl with the successful application of the CWS-2009 treatment protocol.

[Analysis of intraocular lens displacement relative to the haptic plane by ultrasound biomicroscopy data]. / Analiz smeshchenii opticheskoi chasti intraokulyarnoi linzy otnositel'no ploskosti gapticheskikh elementov po dannym ul'trazvukovoi biomikroskopii.

PURPOSE: This study analyzed cases with postoperative displacement of intraocular lens (IOL) relative to the haptic plane using ultrasound biomicroscopy (UBM). MATERIAL AND METHODS: The study analyzed biometry data of 231 patients (277 eyes) aged 72.39±7.77 years, among them 43.25% were males. IOL position and refraction were analyzed at 1, 3 and 6 months after standard phacoemulsification. RESULTS AND DISCUSSION: UBM was performed to analyze the position of the IOL. Stabilization of lens position was observed by the 3rd month of observation. Among the cases with displacement of the optical part relative to the haptic plane, the forward shift was determined in 24.85%, backward - in 16.67% of cases. Eyes with opposite IOL displacements differed significantly in lens diameter, ciliary sulcus diameter and the power of implanted IOL. IOL shift towards the retina produced significant hyperopic refractive error. A discriminant function was compiled using preoperative biometry data comprising the model for predicting backward IOL displacement with high probability. However, we failed to obtain a qualitative model for forward IOL displacements. CONCLUSION: Calculation error in modern formulas could occur because of IOL displacement, including the shift of the optical part relative to the haptic plane in postoperative period. Analysis of biometry data allows calculating IOL displacement towards the retina with a high probability, which could help avoid hyperopic refraction error postoperatively.

Choosing the optimal mandible position for inferior alveolar nerve block (IANB) using finite element analysis.

BACKGROUND: One of the most popular methods of local anesthesia in dentistry, inferior alveolar nerve block (IANB) involves the blockade of the inferior alveolar nerve (IAN) and lingual nerve (LN) in the pterygomandibular space. Despite the large number of works describing the contents of this space, the spatial displacements of the anatomical structures of this area at different positions of the mandible have not been sufficiently studied. The aim of our study was to study the spatial movements of the IAN and inferior alveolar artery (IAA) at various positions of the mandible using computer simulation and finite element analysis to find the safest way to conduct IANB. MATERIALS AND METHODS: Reverse engineering was used to create a model of the cranial base and the mandible based on computed tomography (CT) data obtained from patient N (male, 24 years old), the arteries of the head and neck were designed from the data of multiphase angiography of patient M (female, 61 years old). Masticatory muscles, sphenomandibular ligament, temporomandibular joint and mandibular nerve were modeled in the SolidWorks software package based on an open database of anatomical structures. The finite element grid was generated in the Solidworks software. In the first series of experiments, the displacement of the mandible was modeled along the vertical axis down by 48 mm (maximum opening of the mouth), in the second series, the jaw was displaced vertically by 48 mm with a simultaneous transversal movement of 10 mm, in the third series, the jaw was displaced along the vertical axis down by 34 mm and transversally by 22 mm. RESULTS: The largest distance between IAN and IAA was noted in the third series of experiments. The distance between the nerve and the vessel was minimal in the first series, with an open mouth without lateral displacements. CONCLUSION: The generated computer model opens new possibilities for studying the dynamic anatomy of the pterygomandibular space. The results of this study can be used for further experimental and clinical trials to find the safest approach to the implementation of IANB, as well as applied in the practice of the educational process.

Influence of Extracellular Vesicles from the Follicular Fluid of Young Women and Women of Advanced Maternal Age with Different miRNA Profiles on Sperm Functional Properties.

We studied the effect of co-culturing of extracellular vesicles in the follicular fluid of young women and women of advanced maternal age on sperm motility. Vesicles were obtained by differential centrifugation. The sperm fraction was isolated from the seminal fluid of 18 patients (age 28-36 years). The spermatozoa were incubated with vesicles (1:2 ratio) for 60 or 120 min at 37°C in a CO2 incubator. A fraction of spermatozoa incubated without vesicles served as the control. After the incubation, the sperm samples were sedimented by centrifugation, fixed in 2.5% glutaraldehyde, and analyzed by transmission electron microscopy. RNA was isolated from the follicular fluid vesicles by column method followed by cDNA synthesis in a reaction mixture according to miScript II RT Kit protocol (Qiagen). After 60-min incubation with extracellular vesicles from the follicular fluid of women of advanced maternal age, the sperm motility and hyperactivation slightly changed in comparison with the group where incubation was performed with follicular fluid vesicles from young women and control group. Follicular fluid miRNA profiles in women of different ages varied, which suggests different functional compositions and effects of follicular fluid vesicles of different age groups on sperm characteristics. Transmission electron microscopy revealed differences in the interaction of follicular fluid vesicles from women of different age groups with spermatozoa. Further study of the effect of extracellular vesicles from the follicular fluid and analysis of their transcriptomic, proteomic, and metabolomic composition on sperm mobility and fertilizing ability will improve the effectiveness of assisted reproductive technology programs in patients with male infertility.

[Iron deficiency in Russia heart failure patients. Observational cross-sectional multicenter study].

Aim    To evaluate the prevalence of iron deficiency (ID) in Russian patients with heart failure (HF).Material and methods    Iron metabolism variables were studied in 498 (198 women, 300 men) patients with HF. Data were evaluated at admission for HF (97 %) or during an outpatient visit (3 %). ID was determined according to the European Society of Cardiology Guidelines.Results    83.1 % of patients had ID; only 43.5 % of patients with ID had anemia. Patients with ID were older: 70.0 [63.0;79.0] vs. 66.0 years [57.0;75.2] (p=0.009). The number of patients with ID increased in parallel with the increase in HF functional class (FC). Among patients with ID, fewer people were past or current alcohol users (p=0.002), and a greater number of patients had atrial fibrillation (60.1 vs. 45.2 %, p=0.016). A multiple logistic regression showed that more severe HF (HF FC) was associated with a higher incidence of ID detection, whereas past alcohol use was associated with less pronounced ID. An increase in N-terminal pro-brain natriuretic peptide (NT-proBNP) by 100 pg/ml was associated with an increased likelihood of ID (odds ratio, 1.006, 95 % confidence interval: 1.002-1.011, p=0.0152).Conclusion    The incidence rate of HF patients is high in the Russian Federation (83.1 %). Only 43.5 % of these patients had anemia. The prevalence of ID in the study population increased with increases in HF FC and NT-proBNP.

Cell mechanics can be robustly derived from AFM indentation data using the brush model: error analysis.

The brush model was introduced to interpret AFM indentation data collected on biological cells in a more consistent way compared just to the traditional Hertz model. It takes into account the presence of non-Hertzian deformation of the pericellular brush-like layer surrounding cells (a mix of glycocalyx molecules and microvilli/microridges). The model allows finding the effective Young's modulus of the cell body in a less depth-dependent manner. In addition, it allows finding the force due to the pericellular brush layer. Compared to simple mechanical models used to interpret the indentation experiments, the brush model has additional complexity. It raises the concern about the possible unambiguity of separation of mechanical properties of the cell body and pericellular layer. Here we present the analysis of the robustness of the brush model and demonstrate a weak dependence of the obtained results on the uncertainties within the model and experimental data. We critically analyzed the use of the brush model on a variety of AFM force curves collected on rather distinct cell types: human cervical epithelial cells, rat neurons, and zebrafish melanocytes. We conclude that the brush model is robust; the errors in the definition of the effective Young's modulus due to possible uncertainties of the model and experimental data are within 4%, which is less than the error, for example, due to a typical uncertainty in the spring constant of the AFM cantilever. We also discuss the errors of parameterization of the force due to the pericellular brush layer.

Influence of Extracellular Vesicles of the Follicular Fluid on Morphofunctional Characteristics of Human Sperm.

We studied the effect of extracellular vesicles of the follicular fluid on morphofunctional characteristics of human spermatozoa using CASA (computer-assisted sperm analysis) analytical system. The vesicles were obtained by sequential centrifugation at different rotational speeds and frozen at -80°C in the Sydney IVF Gamete Buffer medium. The sperm fraction was isolated from the seminal fluid of 21 patients aged 27-36 years by differential centrifugation in a density gradient. The precipitate was suspended in Sydney IVF Gamete Buffer to a concentration of 106/ml and incubated with vesicles (1:2) at 37°C in a CO2 incubator for 30 min and 1 h. Sperm fraction incubated without vesicles served as the control. After incubation, some sperm samples were centrifuged at 700g for 5 min and fixed in 2.5% glutaraldehyde in 0.1 M buffer for transmission electron microscopy. After 30-min and 1-h incubation, the progressive and total sperm motility improved, the curvilinear and linear velocity of spermatozoa did not change significantly. Incubation with vesicles significantly changed the trajectory of sperm movement, which can attest to an increase in their hyperactivation and, probably, fertilizing capacity. Analysis of the effect of extracellular vesicles of follicular fluid on sperm motility will help to improve the effectiveness of assisted reproductive technology programs with male infertility factor by improving sperm characteristics in patients with asthenozoospermia and increasing the fertilizing ability of the sperm.

Epinephrine affects gene expression levels and has a complex effect on biofilm formation in M icrococcus luteus strain C01 isolated from human skin.

In this study, the effect of epinephrine on the biofilm formation of Micrococcus luteus C01 isolated from human skin was investigated in depth for the first time. This hormone has a complex effect on biofilms in various systems. In a system with polytetrafluoroethylene (PTFE) cubes, treatment with epinephrine at a physiological concentration of 4.9 × 10-9 M increased the total amount of 72-h biofilm biomass stained with crystal violet and increased the metabolic activity of biofilms, but at higher and lower concentrations, the treatment had no significant effect. On glass fiber filters, treatment with the hormone decreased the number of colony forming units (CFUs) and changed the aggregation but did not affect the metabolic activity of biofilm cells. In glass bottom plates examined by confocal microscopy, epinephrine notably inhibited the growth of biofilms. RNA-seq analysis and RT-PCR demonstrated reproducible upregulation of genes encoding Fe-S cluster assembly factors and cyanide detoxification sulfurtransferase, whereas genes encoding the co-chaperone GroES, the LysE superfamily of lysine exporters, short-chain alcohol dehydrogenase and the potential c-di-GMP phosphotransferase were downregulated. Our results suggest that epinephrine may stimulate matrix synthesis in M. luteus biofilms, thereby increasing the activity of NAD(H) oxidoreductases. Potential c-di-GMP pathway proteins are essential in these processes.

Laser Microtomography for IVF Oocyte of Human.

For in vitro fertilization technology, the quality of oocytes has a direct impact on the egg fertilization and developmental competence of early embryo. The morphological criteria are used for the estimation of oocyte quality before its fertilization in vitro. To date, only one method is known to determine the maturity of oocyte. This is the routine observation with a light microscope. The aim of this article was to adapt the noninvasive quantitative laser scanning microtomography (QLSM) for the investigation of morphological features of a human oocyte in vitro. This approach was used to accumulate the Z-stack gallery of optical sections of IVF oocyte. The layer-by-layer acquisition allows the fine cytoplasmic structure imaging. Applying the QLSM Z-stack of optical sections, the cellular volume was calculated with quantitative 3D reconstruction of a human oocyte. The volume value and intracellular structure were used as novel criteria to assess the oocyte state after the stress evoked by cryopreservation procedure.

Noninvasive Laser Microtomography of a Human Isolated Oocyte.

Using a non-invasive approach, quantitative laser scanning microtomography (QLSM), the morphology of human oocyte was studied layer-by-layer. Then, the cell volume was computed based on 3D reconstruction of a stack of optical sections obtained by QLSM. The integrity of oocyte membrane after cryopreservation was assessed by measuring the changes in oocyte volume in response to hypotonic shock.

Difference in biophysical properties of cancer-initiating cells in melanoma mutated zebrafish.

Despite sharing oncogenetic mutations, only a small number of cells within a given tissue will undergo malignant transformation. Biochemical and physical factors responsible for this cancer-initiation process are not well understood. Here we study biophysical differences of pre-melanoma and melanoma cells in a BRAFV600E/P53 zebrafish model. The AFM indentation technique was used to study the cancer-initiating cells while the surrounding melanocytes were the control. We observed a statistically significant decrease in the modulus of elasticity (the effective Young's modulus) of cancer-initiating cells compared to the surrounding melanocytes. No significant differences in the pericellular coat surrounding cells were observed. These results contribute to a better understanding of the factors responsible for the initiation of cancer.

[Antioxidant potential of wild rose].

The presence in the hips of a complex of biologically active substances causes an antioxidant effect. The aim of this study was to evaluate antioxidant activity of varietal rosehip of the Scientific Research Institute «Zhigulevskiye Sady¼ collection, which was cultivated in Samara region in the season 2016. Material and methods. Antioxidant activity was analysed (antiradical activity, restoring force by FRAP method (Ferric Reducing Antioxidant Power), antioxidant activity in the system of linoleic acid), the content of total phenols and total flavonoids was determined. Along with this, vitamin C content, the total content of organic acids, reducing sugars, soluble substances were determined. Results and discussion. The results of this research indicated high biological activity of the rosehip test samples. Thus, the maximum content of total phenols was found in the following varieties: Samarskiy Yubileyniy (966 mg of gallic acid/100g of feedstock), Samarskiy (922 mg/100g), Desertniy (858 mg/100 g), Krupnoplodniy VNIVI (723 mg/ 100 g). High content of total flavonoids was determined in Desertnyi variety (442 mg of catechin/100g of feedstock) and Rosa spinosissima L. variety (418 mg/100 g). Anthocyan presence was observed only in Rosa spinosissima L. (90.86 mg cyanidin- 3-glycoside/100g of feedstock), as evidenced by its deep purple color. Desertnyi variety showed the highest indicator of antiradical activity (IC50=2.7 mg/сm3) and restoring force (18.18 mol Fe2+/1 kg of feedstock). The ability to inhibit the oxidation of linoleic acid was demonstrated by all the given samples. For the rosehip the content of vitamin C (leading variety: Rosa mollis Sm. - 67.3 mg%, Krupnoplodniy VNIVI - 47.3 mg%), acidity (leading variety: Yubileyniy - 1.39%), mass fraction of reducing sugars (leading variety: Samarskiy - 9.9%), the content of soluble substances (leading variety: Desertniy - 22.2%) were determined. Conclusion. Hence, the given varieties of rosehip have a high antioxidant activity. According to this, it can be recommended for mass cultivation on the territory of Samara region.

Effect of Mechanical Microvibration on the Quality of Human Embryos during In Vitro Culturing and Outcomes of Assisted Reproduction Technologies.

We summarized the data on the role and efficiency of mechanical microvibration during embryo culturing in assisted reproduction protocols. The efficiency of culturing systems with microvibration in assisted reproduction programs and possible mechanisms of microvibration effect on the preimplantation development of human embryos are discussed.

Correction to: Comparative Analysis of the Expression of Glutathione Peroxidase and Glutathione Reductase Genes in Human Sperm after Cryopreservation.

The second author's name should read A. Yu. Romanov.

[Differential diagnosis of dental enamel focal demineralization and fluorosis by means of spectrophotometry]. / Differentsial'naia diagnostika ochagovoi demineralizatsii émali zubov i fliuoroza s pomoshch'iu spektrofotometricheskogo metoda.

The article presents the results of spectrophotometric tooth enamel scanning for differential diagnosis of focal enamel demineralization and fluorosis. Research was conducted in vivo on teeth affected by these diseases. VITA EasyShade spectrophotometer measurements were made on the affected area and on the visually healthy part of enamel. The lightness appeared as the only one differential significant optical characteristics of tooth enamel. Lightness metrics were higher in the case of initial caries than on the healthy part of enamel when these metrics were lower in the case of fluorosis than on the healthy part of enamel.

Comparative Analysis of the Expression of Glutathione Peroxidase and Glutathione Reductase Genes in Human Sperm after Cryopreservation.

We analyzed the effect of cryopreservation on the expression of glutathione peroxidase (GPX1) and glutathione reductase (GSR) genes in human sperm cells (15 sperm samples from fertile donors and 10 samples from infertile patients). The relative expression of GPX1 and GSR genes was determined by real-time PCR. The rate of post-thaw recovery was 2.1 times higher in the group of fertile donors. A significant increase in the expression of GPX1, but not GSR, was observed in sperm samples from infertile patients, while in patients with infertility, GPX1 expression significantly decreased after cryopreservation/thawing, in samples from fertile donors after the same procedure it increased to the level observed in the sperm samples from infertile patients. A positive correlation was revealed between GPX1 expression and sperm cryotolerance.