[Blood eosinophilia in patients with severe bronchial asthma and chronic polypous rhinosinusitis treated with dupilumab: A review].

Bronchial asthma and chronic polypous rhinosinusitis are diseases associated with a T2-inflammatory immune response. These nosologies can be combined, creating the preconditions for a more severe course of multimorbidity, requiring the use of genetic engineering biological therapy. Dupilumab is a monoclonal antibody that can specifically bind to the alpha subunit of the interleukin-4 receptor and block the action of interleukins 4 and 13, which play a key role in the development of T2 inflammation. Numerous studies have demonstrated the high effectiveness of this medicament. The use of dupilumab in some cases may be accompanied by an increase in eosinophils in the blood. This article presents scientific base and our own experience in treating patients with dupilumab-associated eosinophilia, in addition we describe an algorithm for examining this group of patients for the purpose of timely diagnosis of diseases such as eosinophilic granulomatosis with polyangiitis, eosinophilic pneumonia, etc. It should be noted that in the most cases eosinophilia during targeted therapy with dupilumab is temporary and does not cause clinical manifestations.

[Histomorphometric characteristics of age-related changes in cricoid cartilage.]

The paper presents the results of a histomorphometric study of age-related changes in hyaline cartilage tissue by the example of studying cricoid cartilage from 88 corpses of males of various ages. A comparative assessment of the parameters between the groups was carried out and the dynamics of age-related changes in the main ones was studied. As a result of the studies, the greatest correlation with age was recorded in the following indicators: length (r=0,88) and width (r=0,89) of the ossification zone, the area of bone (r=0,87), adipose (r=0,77) and reticular tissue (r=0,86), the number of chondroblasts (r=-0,7). It was found that the transformations of cricoid cartilage occurring with age are characterized by a decrease in the area of cartilage tissue, the average number of chondrocytes and chondroblasts also tends to decrease steadily with increasing age. Age-related bone tissue transformations are characterized by an increase in the area of bone tissue and the transformation of the tissue structure itself: the number of bone beams and their dimensional characteristics (length and width) change. The results of the study allow us to evaluate the age dynamics of quantitative markers of involution processes of hyaline cartilage tissue in men by the example of studying cricoid cartilage and supplement the existing ideas about the pathogenesis of age-associated cartilage diseases.

Molecular fingerprints resolve affinities of Rhynie chert organic fossils.

The affinities of extinct organisms are often difficult to resolve using morphological data alone. Chemical analysis of carbonaceous specimens can complement traditional approaches, but the search for taxon-specific signals in ancient, thermally altered organic matter is challenging and controversial, partly because suitable positive controls are lacking. Here, we show that non-destructive Fourier Transform Infrared Spectroscopy (FTIR) resolves in-situ molecular fingerprints in the famous 407 Ma Rhynie chert fossil assemblage of Aberdeenshire, Scotland, an important early terrestrial Lagerstätte. Remarkably, unsupervised clustering methods (principal components analysis and K-mean) separate the fossil spectra naturally into eukaryotes and prokaryotes (cyanobacteria). Additional multivariate statistics and machine-learning approaches also differentiate prokaryotes from eukaryotes, and discriminate eukaryotic tissue types, despite the overwhelming influence of silica. We find that these methods can clarify the affinities of morphologically ambiguous taxa; in the Rhynie chert for example, we show that the problematic "nematophytes" have a plant-like composition. Overall, we demonstrate that the famously exquisite preservation of cells, tissues and organisms in the Rhynie chert accompanies similarly impressive preservation of molecular information. These results provide a compelling positive control that validates the use of infrared spectroscopy to investigate the affinity of organic fossils in chert.

[Results of modified submandibular surgical approach usage for the mouth floor phlegmons treatment with respect to dynamic of local non-specific resistance]. / Rezul'taty primeneniya modifitsirovannogo podnizhnechelyustnogo operativnogo dostupa dlya lecheniya odontogennykh flegmon dna polosti rta s uchetom dinamiki lokal'noi nespetsificheskoi rezistentnosti.

THE AIM OF THE STUDY: Was increasing of treatment effectiveness of patients with mouth floor odontogenic phlegmon (MFOP) by modified of surgical approach usage. MATERIALS AND METHODS: The prospective controlled, randomized, simple blinded clinical trial, II b level of evidence, comprised 86 patients with MFOP which formed main and control groups. The main group consisted of 40 patients treated with designed «Method for surgical treatment of odontogenic oral phlegmon of mouth floor with partial dissection of sublingual-submandibular sac¼. The control group involved 46 patients treated with traditional surgical procedure. The differences significance between the values has been evaluated with nonparametric Fisher's exact P-test and parametric Student's t-test for independent samples. RESULTS: The modified surgical approach statistically significantly (p<0.05) decreased the number of inflammatory complications (from 26±7% to 9±4%), terms of secondary sutures application (from 7.9±1.4 to 5.7±1.6 days) and time of hospital stay (from 9.4±1.8 to 8.3±1.7 days). On the 5th day of treatment white blood cells count in the main group was significantly (p<0.05) lower (7.2±1.1·109/l) than in controls (9.4±1.3·109/l) showing improved intoxication syndrome resolution in the main group. Index of reaction of bacteria adsorption to the oral epithelium in the main group on the 5th day of treatment was significantly (p<0.05) higher (77.1±6.9%) than in controls (62.4±7.1%). More successful correction of local non-specific resistance was registered in the main group. CONCLUSION: The proposed modified submandibular surgical approach is more effective for the treatment of patients with MFOP.

Controlling the parameters of focused ion beam for ultra-precise fabrication of nanostructures.

At present, the focused ion beam method is an effective technique for nanoscale profiling of a solid surface and prototyping of micro- and nanoscale structures. The article reveals the results of experimental studies on improving the accuracy and resolution of nanoscale profiling of the surface of solids with a focused ion beam. Investigations of the regularities of the influence of the focused ion beam current, beam dwell time and overlap on the parameters of nanoscale structures and the surface profile have been carried out. The influence of the FIB parameters on the deviation of the structure profile from the specified by the template was estimated. Experimental studies have been carried out to determine the influence of the direction of scanning of the ion beam by the template on the magnitude of the error that occurs when the structure of the graphic template is transferred to the substrate. The optimal relationships between the FIB current and the dimensions of the structures being formed have been determined, thus making it possible to ensure the highest accuracy and rate of formation of nanoscale structures. The results can be used to optimize the choice of the ion-beam milling parameters to achieve the maximum accuracy of reproduction of the given sizes of structures.

Osseointegration evaluation of UHMWPE and PEEK-based scaffolds with BMP-2 using model of critical-size cranial defect in mice and push-out test.

It the present study the push-out mechanical test was adopted for mouse model of implantation in critical-size cranial defects to evaluate the effectiveness of implant-skull fusion. As implants, disks of porous ultra-high molecular weight polyethylene (UHMWPE) and polyetheretherketone (PEEK) with hydroxylapatite (HA) with and without loading of recombinant bone morphogenetic protein-2 (BMP-2) were used. Implantation results were evaluated using histology and micro-computed tomography (micro-CT). In the case of both UHMWPE/HA and PEEK/HA, BMP-2 loading resulted in a significant increase in the amount of bone tissue in the implantation area, especially at the edges of the defect, and an increase in the value of BV/TV (bone volume/tissue volume) during micro-CT. There was a high correlation of BV/TV values with the maximum load and elastic modulus measured during the puch-out test. The maximum load values showed good convergence within the groups and are comparable to the maximum load values obtained by other authors in the rat model of implantation in critical-size cranial defects. An adapted push-out test can be used to evaluate the quality of osseointegration of the implanted materials.

Morphological Variability of the Mandible Shape of the Common Vole Microtus arvalis (Rodentia, Arvicolidae) Semi-species from the Hybrid Zone.

The mandible shape of the common vole semi-species from the hybrid zone was studied for the first time using the methods of geometric morphometry. The hybrid specimens were found to display high morphological variability being closer to the parental Microtus obscurus than to M. arvalis form. The main trends of changes in the mandible shape were primarily observed in the horizontal plane.

[Features of hemodynamics in patients with a local form of odontogenic purulent infection and chronic diseases of the cardiovascular system]. / Osobennosti gemodinamiki u bol'nykh s odontogennoi gnoinoi infektsiei na fone khronicheskoi patologii serdechno-sosudistoi sistemy.

The aim of the study was to study hemodynamic parameters in patients with a local form of odontogenic infection (LFOI) in the presence of diseases of the cardiovascular system and without background pathology. 5 groups were formed: group 1 - patients with LFOI without background diseases (49 patients, mean age 29 [25; 37] years); group 2 - healthy individuals (25 people, mean age 24.7±0.76 years); group 3 - patients with LFOI and diseases of the cardiovascular system (34 patients, mean age 51.2±2.37 years); group 4 - patients without LFOI with diseases of the cardiovascular system (25 patients, mean age - 46 [43; 50] years); group of patients with LFOI without background pathology, comparable in age and sex composition with group 3 (group 5, 28 patients, mean age 48.9±2.30 years). The presence of LFOI in the patient in the absence of background pathology determines the heart rate (78 [72; 82] in 1 min) in comparison with the control group of healthy individuals (68.0±0.85 in 1 min), the values of minute volume of blood circulation (5.25±0.14 l/min) and systolic index (2.87 [2.56; 3.35]) l·m-2·min-1. The development of LFOI in patient with hypertension/ischemic heart disease was not associated with increased blood circulation features, such as the increase in resting heart rate values, the values of minute volume of blood circulation and systolic index.

Combined Effect of Bone Morphogenetic Protein-2 and Erythropoietin on Regeneration of Cranial Bone Defects in Mice.

Using mouse model of regeneration of critical size cranial defects, we studied combined effect of 1 and 10 µg of BMP-2 of prokaryotic origin and recombinant erythropoietin (Epostim) injected subcutaneously in the area of bone defect in a total dose of 6000 U/kg. Erythropoietin considerably improved quantitative and qualitative characteristics of the bone tissue in the site of implantation when used in combination with BMP-2 in both concentrations.

Recombinant Human Erythropoietin Proteins Synthesized in Escherichia coli Cells: Effects of Additional Domains on the in vitro and in vivo Activities.

The aim of this work was to compare biological activities of three variants of bacterially expressed human recombinant erythropoietin (EPO) with additional protein domains: 6His-s-tag-EPO protein carrying the s-tag (15-a.a. oligopeptide from bovine pancreatic ribonuclease A) at the N-terminus and HBD-EPO and EPO-HBD proteins containing heparin-binding protein domains (HBD) of the bone morphogenetic protein 2 from Danio rerio at the N- and C-termini, respectively. The commercial preparation Epostim (LLC Pharmapark, Russia) produced by synthesis in Chinese hamster ovary cells was used for comparison. The EPO variant with the C-terminal HBD domain connected by a rigid linker (EPO-HBD) possesses the best properties as compared to HBD-EPO with the reverse domain arrangement. It was ~13 times more active in vitro (i.e., promoted proliferation of human erythroleukemia TF-1 cells) and demonstrated a higher rate of association with the erythropoietin receptor. EPO-HBD also exhibited the greatest binding to the demineralized bone matrix (DBM) and more prolonged release from the DBM among the four proteins studied. Subcutaneous administration of EPO-HBD immobilized on DBM resulted in significantly more pronounced vascularization of surrounding tissues in comparison with the other proteins and DBM alone. Therefore, EPO-HBD displayed better performance with regard to all the investigated parameters than other examined EPO variants, and it seems promising to study the possibility of its medical use.

Synthesis in Escherichia coli and Characterization of Human Recombinant Erythropoietin with Additional Heparin-Binding Domain.

Recombinant human erythropoietin (EPO) with additional N-terminal heparin-binding protein domain (HBD) from bone morphogenetic protein 2 was synthesized in Escherichia coli cells. A procedure for HBD-EPO purification and refolding was developed for obtaining highly-purified HBD-EPO. The structure of recombinant HBD-EPO was close to that of the native EPO protein. HBD-EPO contained two disulfide bonds, as shown by MALDI-TOF mass spectrometry. The protein demonstrated in vitro biological activity in the proliferation of human erythroleukemia TF-1 cell test and in vivo activity in animal models. HBD-EPO increased the number of reticulocytes in the blood after subcutaneous injection and displayed local angiogenic activity after subcutaneous implantation of demineralized bone matrix (DBM) discs with immobilized HBD-EPO. We developed a quantitative sandwich ELISA method for measuring HBD-EPO concentration in solution using rabbit polyclonal serum and commercial monoclonal anti-EPO antibodies. Pharmacokinetic properties of HBD-EPO were typical for bacterially produced EPO. Under physiological conditions, HBD-EPO can reversibly bind to DBM, which is often used as an osteoplastic material for treatment of bone pathologies. The data on HBD-EPO binding to DBM and local angiogenic activity of this protein give hope for successful application of HBD-EPO immobilized on DBM in experiments on bone regeneration.

Blister-based-laser-induced-forward-transfer: a non-contact, dry laser-based transfer method for nanomaterials.

We show that blister-based-laser-induced forward-transfer can be used to cleanly desorb and transfer nano- and micro-scale particles between substrates without exposing the particles to the laser radiation or to any chemical treatment that could damage the intrinsic electronic and optical properties of the materials. The technique uses laser pulses to induce the rapid formation of a blister on a thin metal layer deposited on glass via ablation at the metal/glass interface. Femtosecond laser pulses are advantageous for forming beams of molecules or small nanoparticles with well-defined velocity and narrow angular distributions. Both fs and ns laser pulses can be used to cleanly transfer larger nanoparticles including relatively fragile monolayer 2D transition metal dichalcogenide crystals and for direct transfer of nanoparticles from chemical vapour deposition growth substrates, although the mechanisms for inducing blister formation are different.

[Deficit of Magnesium and States of Hypercoagulation: Intellectual Analysis of Data Obtained From a Sample of Patients Aged 18-50 years From Medical and Preventive Facilities in Russia].

The states characterized by pronounced hypercoagulable components (deep vein thrombosis, cardio- and cerebro-vascular pathologies) are caused by multiple pathophysiological factors, including insufficient supply of magnesium (Mg) and other micronutrients. AIM: to present results of analysis of the Institute of Microelements Data Base (IMDB) performed from point of view of interrelationships of Mg deficit and hypercoagulable states in adults treated in medico-preventive facilities of Central, Northwestern, Northern, and Siberian federal districts of Russia. METHODS: The analysis was realized as analysis of data obtained in a cross-sectional study. In the cohort of patients (n=1453) formed from the IMBD adequacy of Mg supply was assessed by magnesium levels in blood plasma (Mg BP) (0.69±0.15 mmol/L) and estimates of daily Mg consumption according to dietary diaries (Mg D) (185±90 mg/day). RESULTS: Mg supply was adequate (Mg BP >0.80 mmol/L, Mg D >300 mg/day) in not more than 6 % of patients. Presence of "Hypercoagulation" label in data base was associated with greater number of chronic diseases (2.3±2.1 and 0.83±0.8 with and without this label, respectively, р=0.0006) and elevated risk of the presence on 4 comorbid pathologies (odds ratio [OR] 18, 95 % confidence interval [CI] 10-25, р=0.0006). Mg deficit (Mg BP.

Deficit of Magnesium and States of Hypercoagulation: Intellectual Analysis of Data Obtained From a Sample of Patients Aged 18-50 years From Medical and Preventive Facilities in Russia.

The states characterized by pronounced hypercoagulable components (deep vein thrombosis, cardio- and cerebro-vascular pathologies) are caused by multiple pathophysiological factors, including insufficient supply of magnesium (Mg) and other micronutrients. AIM: to present results of analysis of the Institute of Microelements Data Base (IMDB) performed from point of view of interrelationships of Mg deficit and hypercoagulable states in adults treated in medico-preventive facilities of Central, Northwestern, Northern, and Siberian federal districts of Russia. METHODS: The analysis was realized as analysis of data obtained in a cross-sectional study. In the cohort of patients (n=1453) formed from the IMBD adequacy of Mg supply was assessed by magnesium levels in blood plasma (MgBP) (0.69±0.15 mmol/L) and estimates of daily Mg consumption according to dietary diaries (MgD) (185±90 mg/day). RESULTS: Mg supply was adequate (MgBP >0.80 mmol/L, MgD >300 mg/day) in not more than 6% of patients. Presence of "Hypercoagulation" label in data base was associated with greater number of chronic diseases (2.3±2.1 and 0.83±0.8 with and without this label, respectively, р=0.0006) and elevated risk of the presence on 4 comorbid pathologies (odds ratio [OR] 18, 95% confidence interval [CI] 10-25, р=0.0006). Mg deficit (MgBP.

Recombinant Human Erythropoietin with Additional Processable Protein Domains: Purification of Protein Synthesized in Escherichia coli Heterologous Expression System.

Three variants of human recombinant erythropoietin (rhEPO) with additional N-terminal protein domains were obtained by synthesis in an Escherichia coli heterologous expression system. These domains included (i) maltose-binding protein (MBP), (ii) MBP with six histidine residues (6His) in N-terminal position, (iii) s-tag (15-a.a. oligopeptide derived from bovine pancreatic ribonuclease A) with N-terminal 6His. Both variants of the chimeric protein containing MBP domain were prone to aggregation under nondenaturing conditions, and further purification of EPO after the domain cleavage by enterokinase proved to be impossible. In the case of 6His-s-tag-EPO chimeric protein, the products obtained after cleavage with enterokinase were successfully separated by column chromatography, and rhEPO without additional domains was obtained. Results of MALDI-TOF mass spectrometry showed that after refolding 6His-s-tag-EPO formed a structure similar to that of one of native EPO with two disulfide bonds. Both 6His-s-tag-EPO and rhEPO without additional protein domains purified after proteolysis possessed the same biological activity in vitro in the cell culture.

Recombinant Human Bone Morphogenetic Protein-2 (rhBMP-2) with Additional Protein Domain Synthesized in E. coli: In Vivo Osteoinductivity in Experimental Models on Small and Large Laboratory Animals.

Recombinant human bone morphogenetic protein-2 with an additional s-tag domain (s-tag-BMP-2) synthesized in E. coli is characterized by higher solubility and activity than the protein without additional s-tag domain, which increases the yield during purification and simplifies protein introduction into the osteoplastic materials. The high osteoinductivity of the demineralized bone matrix with s-tag-BMP-2 was shown on the model of regeneration of cranial defects of a critical size in mice and on the model of implantation of porous titanium matrix into defects of femoral and tibial bones in rabbits.

Two Variants of Recombinant Human Bone Morphogenetic Protein-2 (rhBMP-2) with Additional Protein Domains: Synthesis in an Escherichia coli Heterologous Expression System.

Two variants of recombinant human bone morphogenetic protein-2 (rhBMP-2) with additional N-terminal protein domains were obtained by expression in E. coli. The N-terminal domains were s-tag (15-a.a. oligopeptide from bovine pancreatic ribonuclease A) and lz (leucine zipper dimerization domain from yeast transcription factor GCN4). The s-tag-BMP-2 and lz-BMP-2 were purified by a procedure that excluded a long refolding stage. The resulting dimeric proteins displayed higher solubility compared to rhBMP-2 without additional protein domains. Biological activity of both proteins was demonstrated in vitro by induction of alkaline phosphatase in C2C12 cells, and the activity of s-tag-BMP-2 in vivo was shown in various experimental animal models.

Modern Approaches to Studies of New Osteogenic Biomaterials on the Model of Regeneration of Critical-Size Cranial Defects in Rats.

Osteoinductive characteristics of new osteoplastic materials based on demineralized bone matrix of xenogenic origin with high and controlled degree of purification were studied on the model of regeneration of critical-size cranial defects in rats using modern approaches, including histological analysis, evaluation of morphological parameters of the bone tissue obtained by micro-computed tomography, and estimation of bone tissue growth rate using in vivo fluorochrome label. Demineralized bone matrix and, to a much greater extent, its activated form containing modified recombinant growth factor rhBMP-2 with high content of the dimeric form exhibited osteoinductive activity.

[MORPHOLOGICAL CHARACTERISTICS OF OSSEOINTEGRATION AFTERAPPLICATION OF TITANIUM IMPLANTS WITH BIOACTIVE COATING AND RECOMBINANT BONE MORPHOGENETIC PROTEIN].

Experiments were carried out on 22 albino male Wistar rats to study the morphological peculiarities of osseointegration of titanium grafts with bioactive surface stimulated additionally with bone plastic material "Gamalant-paste-FORTE Plus" containing recombinant human bone morphogenetic protein-2 (rhBMP-2). In 9 rats the implants were placed into femoral bones after local treatment of bone canal with rhBMP-2-containing material. Another 9 animals were implanted but received no treatment, 4 rats formed the group of intact control. Zone of osseointegration was studied 4, 8 and 12 weeks after graft placement using histological and morphometric methods as well as immune histochemistry to demonstrate osteonectin, CD68, MMP-9, and TIMP-1. The study showed that preliminary treatment of bone canal with rhBMP-2-containing material preceding implant placement was accompanied by an additional osteoinductive effect. More intense and outrunning bone formation in the area of osseointegration was observed, together with remodeling and compaction of the contiguous cancellous bone, thus providing the necessary balance between MMP-9 and TIMP-1 with a high level of each factor expression.