Hydrogels Based on Recombinant Spidroin Stimulate Proliferation and Migration of Human Corneal Cells.

The effect of recombinant spidroin (RS) hydrogel (HG) on anterior epithelial cells and keratocytes of the human cornea was studied in vitro. Corneal injuries are highly prevalent in developing countries according to the World Health Organization. Various technologies have recently been proposed to restore the damaged surface of the cornea. Use of biodegradable silk-based materials, including recombinant analogs of the spider silk protein spidroin, is an important avenue of research in the field of wound healing and corneal regeneration. Spidroins are well known for their optimal balance of strength and elasticity. Given their biological compatibility, lack of immunogenicity, and biodegradability, spidroins provide a biomaterial for tissue engineering and regenerative medicine. HGs based on RS rS2/12-RGDS were therefore tested for cytotoxicity toward isolated corneal epithelial cells and keratocytes with regard to possible changes in cell phenotype and migratory activity. A promising outlook and therapeutic potential were demonstrated for RS-based HGs.

Tissue-Engineered Constructions in Biophysics, Neurology and Other Fields and Branches of Medicine.

This paper describes the gangliopexy method, a method for creating a new center of local neurohumoral regulation, based on the formation of new connections discovered between the nervous system and the vascular system. The prospects for the development of this method are studied. At the same time, novel concepts about the cycles of nitric oxide and the superoxide anion radical are introduced. A possible role of these cycles is examined in the protection of cells and the body as a whole against oxidative and nitrosative stress, which develops when (in 5-30% of cases) destructive changes in the displaced ganglion lead to vascular complications and an increased risk of mortality. Mechanisms that can protect nerve cells, prevent the development of destructive changes in these cells and reduce the risk of mortality are also investigated.

AB0 and rhesus blood groups as a risk factor for ARVI COVID-19.

The frequency of AB0 and Rhesus blood groups was studied in 12120 patients with COVID-19, 5180 convalescent plasma donors and 118801 healthy donors from Moscow, Smolensk, Yakutsk, Minsk and Gomel. In infected individuals, the frequency of blood group A was significantly higher than in uninfected individuals (41,54 % and 34,39 % respectively, p<0,05), and the frequency of blood group 0, on the contrary, was significantly lower (27,69 % and 36,71 %, p<0,05). The frequency of blood group A was particularly high among patients who died from ARVI COVID-19 - 45,51 % vs. 34,39 %, p=0,008. In some groups of patients, there was a decrease in the frequency of Rh-negative individuals (2,23 % vs. 8,30 %, p<0,001).

[Developmental dysphasia in children: a comparison of the effectiveness of two modes of peptidergic nootropic therapy]. / Disfaziya razvitiya u detei: sravnenie effektivnosti dvukh rezhimov peptidergicheskoi nootropnoi terapii.

OBJECTIVE: To study the therapeutic efficacy of two treatment modes of peptidergic nootropic medication cortexin in children with developmental dysphasia aged 3-4 years. MATERIAL AND METHODS: Ninety-four children with developmental dysphasia were divided into three groups. In group 1 (27 patients), cortexin was administered once a day intramuscularly for 10 days. After this course, the children were not prescribed drug therapy, a second examination was carried out 2 months after the start of treatment. In group 2 (40 patients), two courses of cortexin were administered at 1-month intervals, and the children were also followed up for two months. Control group 3 (27 patients) did not receive medication, but was also followed up for two months. All the parents were provided with recommendations for the stimulation of speech development in children. Before the study and two months later, speech development was assessed with special scales and questionnaires for parents. RESULTS: The increase ratio of the active vocabulary volume by 2 times or more was observed in group 2 (two courses of cortexin treatment) in 80.0% of patients (significant differences with groups 1 and 3, p<0.001), in 44.4% of group 1 (one course of cortexin), in 22.2% of the control group. Over a two-month period, the increase ratio in the active vocabulary and the number of uttered phrases in group 2 was 2.8 and 4.2 times, in group 1 2.3 and 3.6 times, respectively, in the control group it was only 1.4 and 1.5 times. Furthermore, the volume of active vocabulary in group 2 (42.4±3.6) became significantly larger (p=0.01) than in group 1 (31.7±5.6), although its initial values in group 1 (13.7±1.8) and group 2 (14.9±1.7) were similar. CONCLUSION: The results of the study confirm the higher effectiveness of two courses of the peptidergic nootropic medication cortexin in the pharmacotherapy of developmental dysphasia in children, aged 3-4 years, conducted over two months, compared with the indication of one treatment course.

Recombinant Spidroin Films Attenuate Individual Markers of Glucose Induced Aging in NIH 3T3 Fibroblasts.

The effect of bioresorbable materials on aging in cultured mouse NIH 3T3 fibroblasts treated with elevated glucose concentration was investigated. The cells were grown on films produced from the silkworm fibroin and rS1/9, a recombinant analog of Nephila clavipes spidroin 1. Exposure to 50 mM glucose of the cells grown on uncoated glass support resulted in the cell growth retardation. The average areas of the cells and nuclei and the percentage of apoptotic cells increased, whereas the amount of soluble collagen decreased. In contrast, on the fibroin and spidroin films, the cell density and the percentage of 5-bromo-2'-deoxyuridine (BrdU)-positive cells were higher vs. the cells grown on the glass support. The films protected NIH 3T3 fibroblasts from the glucose-induced death. The most prominent effects on the cell density, BrdU incorporation, and apoptosis prevention were observed in the cells cultured on spidroin films. Unlike the cells grown on glass support (decrease in the soluble collagen production) or fibroin (no effect), production of soluble collagen by the cells grown on spidroin films increased after cell exposure to 50 mM glucose. Molecular analysis demonstrated that 50 mM glucose upregulated phosphorylation of the NFκB heterodimer p65 subunit in the cells grown on the glass support. The treatment of cells grown on fibroin films with 5.5 mM or 50 mM glucose had no effect on p65 phosphorylation. The same treatment decreased p65 phosphorylation in the cells on the spidroin films. These results demonstrate the anti-aging efficacy of biomaterials derived from the silk proteins and suggest that spidroin is more advantageous for tissue engineering and therapy than fibroin.

[Pharmacotherapy of psychomotor developmental delay in 6-12 months preterm infants with hypoxic-ischemic encephalopathy (the double-blind comparative multicenter placebo-controlled study)]. / Farmakoterapiia zaderzhki psikhomotornogo razvitiia u detei 6-12 mes, rozhdennykh nedonoshennymi i perenesshikh gipoksicheski-ishemicheskoe porazhenie golovnogo mozga (dvoinoe slepoe sravnitel'noe mnogotsentrovoe platsebo-kontroliruemoe issledovanie).

AIM: To evaluate the efficacy and safety of hopantenic acid (Pantogam) in the complex treatment of prematurely born infants, aged 6-12 months, with psychomotor developmental delay due to hypoxic-ischemic encephalopathy. MATERIAL AND METHODS: Eighty-seven patients were randomized into two groups: 44 received standardized treatment and pantogam for two months, 43 standardized treatment and placebo. Pantogam (syrup 100 mg/ml) or placebo were prescribed orally 15-30 minutes after feeding, twice a day, in a daily dosage of 30-50 mg/kg body weight. The assessment of psychomotor development from birth to two years was performed with the Griffiths Mental Development Scales (GMDS-ER) twice (before and after completion of therapy). RESULTS: The response to two month therapy determined as the reduction of developmental delay for more than 6% of the initial GMDS-ER general quotient (GQ) score was significantly better in the group I after pantogam treatment (63.6% of patients) compared to group II (36.4%, p=0.021). Group I demonstrated the significant decrease of the developmental delay in two domains ('Personal-Social' and 'Performance') and a trend to overcome the delay in three other domains: 'Locomotor', 'Hearing and Speech', 'Eye and Hand Coordination'. The improvement after pantogam treatment was more obvious in the subgroup of infants born late preterm (gestational age 34-36 weeks) compared to infants born moderate preterm (gestational age 32-33 weeks). The favorable safety profile of pantogam was confirmed, comparable to that of placebo. CONCLUSION: Pantogam is efficient and safe medication in the complex treatment of psychomotor developmental delay in preterm infants, aged 6-12 months.

[Potential of peptidergic nootropic therapy in developmental dysphasia in children]. / Vozmozhnosti peptidergicheskoi nootropnoi terapii pri disfazii razvitiia u detei.

AIM: To study the therapeutic efficacy of peptidergic nootropic medication Cortexin during two-month follow-up in children with developmental dysphasia aged 3-4 years. MATERIAL AND METHODS: Fifty-four children with developmental dysphasia were randomized into two equal groups. The cortexin group received the drug once daily intramuscularly during 10 days. After this course, children did not receive any pharmacotherapy and were examined two months after the beginning of treatment. The control group did not receive pharmacotherapy and was examined twice in two months interval. All the parents were provided with recommendations for the stimulation of speech development in dysphasic children. Before treatment and two months later, speech development was assessed by special scales and questionnaires for parents. RESULTS AND CONCLUSION: After treatment with Cortexin, a significant improvement was achieved not only on the 'expressive language' scale but also on the 'attention to speech' and 'impressive language' scales. The volume of active vocabulary was increased by 2.3 times, the number of articulated phrases by 3.6 times. The evaluation of parents' questionnaires demonstrated the decrease of cerebrasthenic, psychosomatic problems, motor clumsiness, hyperactivity, attention deficit and problems with emotional control in children. The therapy contributed to the reduction of difficulties in communication and social adaptation. The positive effect of Cortexin was observed after the completion of treatment (the protracted effect).

[Neurological and neurodevelopmental disorders in preterm-born children (with extremely low, very low or low body weight)]. / Nevrologicheskie narusheniia i rasstroistva psikhicheskogo razvitiia u detei, rozhdennykh nedonoshennymi (s ékstremal'no nizkoi, ochen' nizkoi i nizkoi massoi tela).

AIM: To evaluate the incidence of neurological diseases and neurodevelopmental disorders in preterm-born children, aged 5-8 years, with extremely low (ELBW), very low (VLBW) or low body weight (LBW) at birth. MATERIAL AND METHODS: One hundred and twenty-two preterm-born children, including 36 born with ELBW, 36 born with VLBW and 50 born with LBW, were examined in the age from 5 years 0 months to 8 years 0 months. Presenting complaints were assessed by means of the structured parents' questionnaire. Diagnosis of the nervous system diseases was based on the criteria of ICD-10 and DSM-V. RESULTS AND CONCLUSION: Based on the assessment results, there was no any neurological or neurodevelopmental disorder in 8.3% (n=3) of children with ELBW, 16.7% (n=6) with VLBW, 22.0% (n=11) with LBW at birth. Compared to the general pediatric population, more preterm-born children with ELBW, VLBW or LBW, aged 5-8 years, had developmental dyspraxia, chronic motor tics, tension type headaches and enuresis. Moreover, the higher incidence was found for neurodevelopmental disorders, including attention deficit hyperactivity disorder, autism spectrum disorders, specific learning disabilities, compared with population rates. All conditions were more prevalent in boys than in girls, with the exception of tension type headaches. The frequency of disorders was inversely correlated with gestational age and body weight. However, the results demonstrate reserve capacities of the developing brain in children born preterm and suffered early brain damage.

[Neurodevelopment of children born very preterm with extremely low and very low body weight]. / Nervno-psikhicheskoe razvitie detei, rodivshikhsia gluboko nedonoshennymi s ékstremal'no nizkoi ili ochen' nizkoi massoi tela.

AIM: To evaluate the characteristics of neurodevelopment by the age of 5-8 years in children who were born very preterm with extremely low body weight (ELBW) and very low body weight (VLBW). MATERIAL AND METHODS: Seventy-two patients, aged from 5 years to 8 years were examined. Patients were divided into group I (36 patients born preterm with ELBW (16 boys, 20 girls)) and group II (36 patients born with VLBW (16 boys, 20 girls)). The control group included 30 healthy peers (16 boys, 14 girls). All children were assessed by means of the Griffiths Mental Development Scales, Extended Revised: 2 to 8 years (GMDS-ER 2-8). RESULTS: The value of general quotient (GQ) in patients born with ELBW (73.4±2.1) was significantly lower compared to their healthy peers (80.9±2.1; p=0.036). The same tendency was found in patients born with VLBW who's GQ was decreased by 73.1±3.0 (p=0.101). Concurrently the patients demonstrated lower scores on all six scales, which achieved significant difference with the controls for the 'Locomotor' and 'Performance' scales in group I, and for the 'Performance' and 'Practical reasoning' scales in group II. The boys born with ELBW tended to demonstrate lower results on all six scales and received significantly lower scores on the 'Locomotor' and 'Language' scales. The boys born with VLBW demonstrated significantly lower scores on the 'Language', 'Eye and hand co-ordination' and 'Practical reasoning' scales. In girls significantly lower score on the 'Performance' scale only was shown for those, who were born with ELBW. CONCLUSION: Early diagnosis and characterization of developmental delays in children born very preterm determine the effectiveness of therapeutic measures based on the individual approach and comprehensive medical-psychological-pedagogical support.

Recombinant 1F9 spidroin microgels for murine full-thickness wound repairing.

The study of the stimulating effect of the microgels (MGs) based on recombinant 1F9 spidroin on the regeneration of the deep skin wound in mice was carried out. The use of spidroin MGs was shown to increase significantly the quality of healing compared to the control. The introduction of the MG in the wound edges led to recovery of all the structural elements of the skin: the epidermis, the dermis, including vascular and nervous network, in the periphery of the wound underlying muscles, and skin appendages (sebaceous and sweat glands and hair follicles) was revealed.

Novel 3D-microcarriers from recombinant spidroin for regenerative medicine.

Microcarriers generated from recombinant spidroin 1F9 are suitable for use as an injection material. The microcarriers were a heterogeneous mixture of microgel particles ranging from 50 to 300 µm in size with the predominance of particles of 50-150 µm. The surface of these microparticles had a complex topography and ensured efficient cultivation of primary and immortalized fibroblasts. Intradermal injections of microgel suspensions into the area of full-thickness skin wounds did not lead to the development of acute inflammation in mice; instead, they accelerated the recovery of skin tissue and stimulated neurogenesis and angiogenesis.

[Clinical effectiveness of the infusion therapy under control of the transesophageal dopplerography in acute period of severe combined trauma].

In 72 patients with combined trauma the impact of volume and consistence of infusion therapy on severity of the disease, frequency and severity of infectious complications, duration of MV and ICU stay was assessed. The patients were divided into 2 groups depending on the volemic status control method and infusion algorithm. The main group (35 patients) was controlled by transesophageal dopplerography Cardio Q apparatus ("Deltex Medical", GB) and the infusion therapy was carried out under the control of stroke volume and Ftc. In the control group (37 patients) the volemic status was assessed clinically: BP, CVP, HR, diuresis. The volume of the infusion therapy during the first 12 hours in the main group was significantly higher than in the control group which proved that patients were suffering from hypovolemia, which was not diagnosed by traditional clinical criteria. The ICU stay in the main group was significantly shorter compared to the control group 15.3 +/- 8.2 and 29.5 +/- 10.4 days respectively. Infectious complications occurred in 12 patients out of 35 in main group and 25 out of 37 in the control group. The conclusion of this study is that infusion therapy control with central hemodynamic parameters can shorten the MV time and ICU stay an lower the rate of infectious complications in patients with combined trauma. A mortality decrease in patients with transesophageal dopplerography controlled infusion is not shown.

[Current technologies and changing the medical paradigm in aortic ruptures (analysis of clinical cases)].

The paper describes two clinical cases of aortic rupture of traumatic and nontraumatic genesis, which was difficult to diagnose due to evident neurological symptoms. The analysis of these cases and the data available in the literature leads to the conclusion that aortic rupture is not a nosological entity with a known fatal outcome. Timely diagnosis based on widely practiced up-to-date ultrasound and radiological methods allows one to perform surgical or endovascular treatment in good time, by showing good long-term outcomes.

[Effect of hyperosmolar solutions on partial pressure of oxygen in arterial blood].

The study was undertaken to define the effect of hyperosmolar solutions on arterial blood oxygen levels. Analysis of arterial blood oxygenation after hyperosmolar solution infusions revealed a drop in partial pressure of oxygen (132.14 +/- 70.36, 120.0 +/- 66.48, and 119.24 +/- 70.10 mm Hg). At the same time there was a reduction in the extravascular lung water index (p < 0.05) as compared with the baseline values (7.93 +/- 2.17, 7.31 +/- 1.7), and Z62 +/- 1.99 ml/kg), respectively. This effect is particularly marked in a group of patients in whom the arterial blood content of oxygen is in the lower normal range. To evaluate and eliminate the above effect in reducing the arterial blood level, it is necessary to estimate respiratory indices during hyperosmolar solution infusion.

[Quaternary structure formation by recombinant analogues of spider silk].

A study has been conducted on the morphology of artificial spider silk fibers, prepared from recombinant analogues of spiridons 1 and 2. It has been shown that by stretching out the "as spun" fiber, a reorganization of its spongy matrix occurs, which leads to the formation of microfibrills, followed by a reduction of the diameter of the fiber. The durability of an artificial fiber depends on the degree of stretching and on the substructure of the microfibrills. The model process of artificial fibers preparation reproduces to the great detail the natural process of spider web spinning. Future applications of this model include production of biomaterials with unique properties.

[Use of albumin in infusion-transfusion therapy for acute brain injury].

The investigation is aimed at determining whether it is necessary to include 20% albumin preparations into infusion-transfusion therapy for acute intracranial hemorrhage. An insignificant improvement of central hemodynamic parameters was shown in a group wherein 20% albumin solution was used. At the same time there was a significant increase in the pulmonary extravascular water index as compared with a control group (10.1 +/- 0.5 and 9.0 +/- 0.22 ml/kg, respectively; with no differences in the severity of lung injury, LIS was 0.71 +/- 0.23 scores; while in the group receiving no albumin solution it was 0.65 +/- 0.06 scores. Thus, the routine use of 20% albumin preparation as part of infusion-transfusion therapy for acute intracranial hemorrhage is not effective and rather safe.

[Left helix of polyproline II type and genesis of beta-structures in spidroins 1 and 2 and their recombinant analogs].

The distribution of secondary structures along the polypeptide chains of spider proteins spidroins 1 and 2 and their recombinant analogs has been studied by statistical methods. It was found that these proteins in the monomolecular form contain only trace amounts of beta-structures. At the same time, the regions of the sequence including Ala and Gly are predicted as helical-containing (with alpha-helices and left-helices of polyproline II type). An analysis of literature data and our data obtained in this study shows that the main conformation of the polypeptide chain solutions of spidroins 1 and 2 and their recombinant analogs in water solutions is the left-helix of polyproline II type with some contaminations of alpha-helices and a very small share of beta-structures. The transition to the state with extended conformations, which are peculiar to mature filaments of spider webs, requires the dehydration of the polypeptide chain backbone. Thus, the genesis of beta-structure in spider web proteins is determined by the conditions of conformation transitions between the main regular conformations of the polypeptide chain backbone.

[The problem of osteopenia in a iodine-deficit area].

The aim of the study was to evaluate the condition of bone system in 335 subjects with thyroid gland dysfunction (TGD), residing in iodine deficit areas. Among the subjects there were 116 individuals with thyrotoxicosis (TT), and 219--with hypothyroidism (HT). The control group included 200 practically healthy people of comparable age and sex. The examination included: evaluation of pain syndrome intensity using a 4-point scale; revealing of atraumatic fracture background; measurement of serum levels of thyroid stimulating hormone (TSH) andfree thyroxin by immune-enzyme assay; measurement of bone mineral density (BMD) by dual-energy x-ray absorptiometry using a Dexa-Scan DX-10 device (Direx, Israel) with evaluation of the results according to 1994 WHO recommendations. 79.3% of TT patients and 68% of HT patients suffered from pain syndrome. The difference in atraumatic fracture rate between the groups was insignificant. The mean BMD values in subjects with TGD dysfunction corresponded to osteopenia, while in the control group they were within normal limits. The difference in osteopenic syndrome rate between TT and HTpatients was insignificant, although osteopenia was revealed in 25% of TT patients. The frequency of BMD decrease grew with age; however, this parameter was higher in all groups of patients with TGD vs. control. The most prone to osteopenia are postmenopausal women regardless of thyroid dysfunction character; difference in BMD decrease frequency between men and women with preserved menstrual function, was insignificant. Osteopenic syndrome was more frequent in men with TGD then in healthy men. Thus, in residents of iodine deficit areas TGD is a factor of BTMD decrease.