[Correlation of the Karolinska sleepiness scale with performance variables of the monotonous bimanual psychomotor test]. / Korrelyatsiya Karolinskoi shkaly sonlivosti s povedencheskimi pokazatelyami monotonnogo bimanual'nogo psikhomotornogo testa.

OBJECTIVE: To assess the objectivity of measuring the level of sleepiness in the subjects using a monotonous psychomotor bimanual tapping test developed by us, performed on mobile devices running Android OS. MATERIAL AND METHODS: Four hundred and ninety-four hour-long experiments with the performance of a psychomotor test were conducted on 102 students. Using the method of mixed linear models, correlations between the levels of sleepiness according to the Karolinska Sleepiness Scale (KSS) and the Epworth Sleepiness Scale (ESS) and the behavioral indicators of the test were evaluated. RESULTS: Statistically significant correlations between the increase in KSS scores and such indicators as a decrease in the total number of button taps and an increase in the frequency of «microsleep¼ episodes are shown. Statistically significant correlations of ESS score characteristics with the behavioral indicators of the test were not found. CONCLUSION: A large statistical material shows a reliable correlation of the parameters of the psychomotor test with the level of sleepiness on the Karolinska scale, which allows using the mobile application developed by us to determine the current level of sleepiness /alertness in the field.

Analysis two types of K complexes on the human EEG based on classical continuous wavelet transform.

In our work, we compare EEG time-frequency features for two types of K-complexes detected in volunteers performing the monotonous psychomotor test with their eyes closed. Type I K-complexes preceded spontaneous awakenings, while after type II K-complexes, subjects continued to sleep at least for 10 s after. The total number of K-complexes in the group of 18 volunteers was 646, of which of which type I K-complexes was 150 and type II K-complexes was 496. Time-frequency analysis was performed using continuous wavelet transform. EEG wavelet spectral power was averaged upon several brain zones for each of the classical frequency ranges (slow wave, δ, θ, α, ß1, ß2, γ bands). The low-frequency oscillatory activity ( δ-band) preceding type I K-complexes was asymmetrical and most prominent in the left hemisphere. Statistically significant differences were obtained by averaging over the left and right hemispheres, as well as projections of the motor area of the brain, p<0.05. The maximal differences between the types I and II of K-complexes were demonstrated in δ-, θ-bands in the occipital and posterior temporal regions. The high amplitude of the motor cortex projection response in ß2-band, [20;30] Hz, related to the sensory-motor modality of task in monotonous psychomotor test. The δ-oscillatory activity preceding type I K-complexes was asymmetrical and most prominent in the left hemisphere may be due to the important role of the left hemisphere in spontaneous awakening from sleep during monotonous work, which is an interesting issue for future research.

[Autoimmune hearing loss and a chance of its development in children: literature review and own observations]. / Autoimmunnaya tugoukhost' i veroyatnost' ee formirovaniya v detskom vozraste: obzor literatury i sobstvennye nablyudeniya.

Autoimmune sensorineural hearing loss (AiSNHL) is an uncommon auditory disorder characterized by rapidly progressive bilateral hearing loss and a positive clinical response to treatment with corticosteroids and cytostatics. The prevalence of the disease in the adult population is less than 1% among all cases of subacute and permanent sensorineural hearing loss (precise data are unknown), it is even rarer in children. AiSNHL can be primary (isolated, organ-specific) or secondary (manifestation of another systemic autoimmune disease). The pathogenesis of AiSNHL is based on the proliferation of autoaggressive T cells and the pathological production of autoantibodies to the protein structures of the inner ear, which leads to damage to various parts of the cochlea (possibly also to the retrocochlear parts of the auditory system), less frequently to the vestibular labyrinth. Pathologically, the disease is most often represented by cochlear vasculitis with degeneration of the vascular stria, damage to hair cells and spiral ganglion cells, and endolymphatic hydrops. In 50% of cases, the result of autoimmune inflammation may be fibrosis and/or ossification of the cochlea. The most characteristic symptoms of AiSNHL at any age are episodes of sudden progression of hearing loss, fluctuations of hearing thresholds, and bilateral, often asymmetric impairments. The article presents contemporary ideas of the clinical and audiological manifestations of AiSNHL, the possibilities of diagnosing and treating the disease, and highlights the current approaches to (re)habilitation. Along with literature data, two own clinical cases of an extremely rare pediatric AiSNHL are given.

Improving the efficacy of potato clonal micropropagation by inoculation with the rhizosphere bacteria Azospirillum baldaniorum Sp245 and Ochrobactrum cytisi IPA7.2.

Sustainable development of agriculture depends on the provision of quality seeds to the market. Inoculation with plant-growth-promoting rhizobacteria in in vitro culture can be used to improve the growth eff icacy and performance of microplants. We examined the effect of in vitro inoculation of microplants of the cultivars Nevsky and Kondor with the strains Azospirillum baldaniorum Sp245 and Ochrobactrum cytisi IPA7.2 separately and in combination. We examined the morphological variables of plant growth in in vitro culture and under ex vitro adaptation conditions; we also investigated the growth and performance of the plants in the greenhouse. The dependence of the inoculation eff icacy on potato genotype, growth stage, and inoculum composition was ascertained throughout the experiment. In vitro, A. baldaniorum Sp245 alone and in combination with O. cytisi IPA7.2 promoted the formation of roots on the microplants of both cultivars and the growth of Nevsky shoots. During plant growth ex vitro, all growth variables of the Nevsky microplants were promoted by O. cytisi IPA7.2 alone and in combination with A. baldaniorum Sp245. In both cultivars grown in the greenhouse, shoot growth was promoted in most inoculation treatments. The survival ability of the Nevsky microplants in the greenhouse increased 1.7-fold under the effect of simultaneous inoculation. Inoculation of microplants with a combination of A. baldaniorum Sp245 and O. cytisi IPA7.2 increased the number of Nevsky minitubers 1.5-fold and the number of Kondor minitubers 3.5-fold. Inoculation with the tested strains can be used to promote the growth of microplants and increase the yield of minitubers in potato seed breeding for the production of healthy planting material.

[Interhemispheric asymmetry of EEG rhythm connections during spontaneous awakenings after short sleep episodes during a monotonous psychomotor test]. / Mezhpolusharnaya asimmetriya svyazei ritmov EEG pri spontannykh probuzhdeniyakh posle kratkovremennykh epizodov sna pri vypolnenii monotonnogo psikhomotornogo testa.

OBJECTIVE: To study interhemispheric asymmetry (IHA) according to electroencephalography (EEG) data of a healthy person during cognitive awakening from the second stage of daytime sleep before restoring the performance of the psychomotor test. MATERIAL AND METHODS: In 23 healthy adult subjects, we studied IHA in the amplitude-amplitude interaction of EEG rhythms for 20 sec segments before spontaneous awakening determined by the moment the alpha-rhythm appearance on the EEG and the subsequent onset of psychomotor activity. The state of the subject during this period (in the initial stage of the so-called cognitive awakening preceding the behavioral awakening), when the person is unable to move, but is able to perceive external stimuli, is an experimental model for highlighting signs of conscious activity of patients when coming out of a coma. Wavelet transform was used to calculate the rhythmic characteristics of bioelectrical activity. The Kendall correlation coefficient served as a measure of rhythm interaction. RESULTS: IHA in the interaction of EEG rhythms is dynamic nature and formed by theta rhythm connections with alpha2- and beta-rhythms in the left brain hemisphere and delta-type connections in the right. CONCLUSION: Possibly, the greater activation of the left hemisphere is related to the retrieval of an instruction from memory, which subsequently allows to return to the activity interrupted by sleep.

[Effects of weak low-frequency electromagnetic field on sleep structure during daytime sleep]. / Vliyanie ekspozitsii slabogo elektromagnitnogo polya sverkhnizkoi chastoty na strukturu dnevnogo sna.

OBJECTIVE: To test the hypothesis that weak electromagnetic fields of low frequencies (0.5-26 Hz) could affect daytime sleep features and structure. MATERIAL AND METHODS: Parameters of daytime sleep continuity were compared in the study with counterbalanced control/exposition (40 min exposure to electromagnetic field at 1 Hz/0.004 µT) scheme in 22 healthy volunteers. Nonlinear regression model was used to assess daytime sleep continuity. RESULTS: Exposure to a weak electromagnetic field of ultra-low frequency significantly improved the quality of sleep, assessed by the indicator of sleep continuity, namely, there were fewer transitions from the second and deeper stages of sleep to the first stage and to the state of wakefulness (p<0.0001). CONCLUSION: The results can be used to develop non-pharmacological methods of sleep correction, as well as to improve the quality of short-term sleep and its positive effect on well-being, cognitive function and working capacity.

A FAMILY HISTORY OF DUCHENNE MUSCULAR DYSTROPHY.

The purpose of the study is to examine in depth and analyze the clinical and some paraclinical characteristics for a family history of Duchenne muscular dystrophy. We analyzed the follow up clinical and laboratory data of Duchenne muscular dystrophy in two brothers-german, aged 16 and 14, respectively. The patients underwent a standardized examination, involving studying the medical case history, general clinical data, determining Sheldon's somatotype and the constitutional type, the detailed neurological status examination, testing a personality type, laboratory and instrumental examinations. Through the laboratory examination we determined the general blood test indicators, total serum protein levels, total cholesterol, the ALAT, ASAT, CPK levels, the indicators of the immunogram, myositis profile and the genetic markers of the disease. The instrumental examination included the ultrasound of the abdominal organs, muscles, as well as echo-cardiography, electroneuromyography. A complete examination fragment of 42 patients with myodystrophies is presented. The paper presents the neurological examination results, the genetic study data and the CPK level indicators in the representatives of Duchenne muscular dystrophy family history. The given family history of Duchenne muscular dystrophy showed two brothers-german to have differences both in the defective dystrophin gene exons at Xp21 and in the disease clinical picture. Thus, patient A., who is an elder brother was detected to have exon 47, 48, 50 and 52 deletion, and patient B., who is a younger brother, was found to have exon 45-43 deletion. The presented family history of Duchenne muscular dystrophy acknowledges the fact that the clinical, genetic, biochemical and other characteristics in patients with dystrophinopathies warrant further comprehensive investigations in order to update diagnostic and prognostic techniques, considering the great medical and social significance of this disabling pathology. However, the onset age of the disease, the clinical course, and the changes in the CPK level were different. Due to the muscle ultrasound both patients were detected to have degenerative changes in the proximal upper and lower limbs.

Profiling of non-criteria antiphospholipid antibodies in patients with SLE: differentiation of thrombotic SLE patients and risk of recurrence of thrombosis.

To reveal the clinical significance of criteria and non-criteria antiphospholipid antibodies detected by line immunoassay in comparison with ELISA, systemic lupus erythematosus patients with and without thrombotic events were investigated. Thus, 107 systemic lupus erythematosus patients (48% with deep vein thrombosis or/and arterial thrombosis) and 120 healthy donors were enrolled. Serum antiphospholipid antibodies were detected by ELISA (Orgentec Diagnostika, Germany) and line immunoassay (GA Generic Assays, Germany). Lupus anticoagulant and IgG to cardiolipin and ß2GPI but not IgM as well as triple positivity by ELISA and line immunoassay were linked with thrombosis in systemic lupus erythematosus. IgG to phosphatidylinositol and phosphatidylserine by line immunoassay showed significantly higher levels in systemic lupus erythematosus with deep vein thrombosis/arterial thrombosis than without and were independent risk factors for deep vein thrombosis (odds ratio 3.9, 95% confidence interval 1.1, 13.2) and arterial thrombosis (odds ratio 5.1, 95% confidence interval 1.3, 19.8) as well as thrombosis (odds ratio 3.6, 95% confidence interval 1.1, 11.3) and recurrence thereof (odds ratio 6.9, 95% confidence interval 2.1, 22.6), respectively. The occurrence of >4 IgG antiphospholipid antibodies by line immunoassay was an independent risk factor for thrombosis (odds ratio 10.9, 95% confidence interval 1.2, 101.5), arterial thrombosis (odds ratio 14.6, 95% confidence interval 2.5, 86.3), deep vein thrombosis (odds ratio 5.8, 95% confidence interval 1.0, 32.4) and recurrence of thrombosis (odds ratio 35.9, 95% confidence interval 3.8, 342.8). Line immunoassay is a promising multiplex test for the simultaneous detection of criteria and non-criteria antiphospholipid antibodies. Profiling of antiphospholipid antibodies by line immunoassay can differentiate systemic lupus erythematosus patients with thrombosis from systemic lupus erythematosus patients without and assess the risk for thrombosis and recurrence thereof.

RENAL, HEPATIC AND IMMUNE FUNCTION INDICES IN PATIENTS WITH DUCHENNE MUSCULAR DYSTROPHY.

The purpose of the study is to examine in depth and analyze renal, hepatic and immune function indices in patients with Duchenne muscular dystrophy. We analyzed the follow up clinical and laboratory data of Duchenne muscular dystrophy in 32 patients. The patients underwent a standardized examination, involving studying the medical case history, general clinical data, determining Sheldon's somatotype and the constitutional type, the detailed neurological status examination, testing a personality type, laboratory and instrumental examinations. Through the laboratory examination we determined the general blood test indicators, total serum protein levels, total cholesterol, the ALAT, ASAT, CPK levels, creatinine and urea blood levels, glomerular filtration rate (GFR), the immunogram indices (dynamic data (B-lymphocytes (CD19/CD45), %; T-lymphocytes (CD3/CD 45), %; T-helpers (CD3/CD45/CD4), %; T-suppressors (CD3/CD45/ CD8), %; CD4:CD8 ratio; natural killer cells ratio, myositis profile (Mi-2, IgG antibodies (idiopathic myositis marker); Ku, IgG antibodies (sclerodermia and myositis combination marker); the PM-Scl complex, IgG antibodies (sclerodermia marker); histidyl tRNA synthetase (Jo-1), IgG antibodies; threonyl-tRNA synthetase (PL-7), IgG antibodies; alanyl-tRNA synthetase (PL-12), IgG antibodies; RING-type E3 ubiquitin-ligase (Ro-52), IgG antibodies and the genetic markers of the disease. The instrumental examination included the ultrasound of the abdominal organs, muscles, as well as echo-cardiography, electroneuromyography. According to the analysis of the immunograms, the T-helpers level was found to be below the reference value in 14 patients (45,2%). Herewith, the B-lymphocytes count was observed to deviate from the norm in only 4 patients (8%). However, there was established the presence of an indirect moderate correlation between the B-lymphocytes count and Creatine phosphokinase level in patients with Duchenne muscular dystrophy; the correlation coefficient was equal to - 0.42 (р<0,05). Having analyzed the renal function indices, we revealed that the creatinine blood level was lower than the lower limit of the reference values in almost a third of patients (21%), but the glomerular filtration rate (GFR) by the SKD-EPI formula was found to be insignificantly reduced in 16% of patients, and it was within 66-87 ml/min/1.73 m2. Regardless, no correlation between the Creatine phosphokinase level, on one side, and creatinine blood level and the glomerular filtration rate, on the other side, was found. However, there was established the presence of an indirect weak correlation between the Creatine phosphokinase level and serum urea level in patients with Duchenne muscular dystrophy (the correlation coefficient was equal to - 0.28, p<0.05). Having analyzed the hepatic function indices, we revealed that such indicators of liver enzymatic activity as alanine aminotransferase and aspartate aminotransferase were significantly over the limit in almost all the patients (94%) with Duchenne muscular dystrophy. Concurrently, we revealed the presence of a direct strong correlation between the Creatine phosphokinase level and the alanine aminotransferase level, which was equal to +0,86 (р<0,05) and the presence of a direct moderate correlation between the aspartate aminotransferase and the Creatine phosphokinase level which was equal to + 0.56 (р<0,05). We also found that the level of alkaline phosphatase was also significantly higher than the normal one in 69% of our patients. The patients with Duchenne muscular dystrophy had various multidirectional disorders of the immune status, impaired renal function (in particular, a decrease in serum creatinine concentration and reduced glomerular filtration rate), as well as the divergence of liver enzyme parameters (in particular, a significant increase in transaminase levels).

[Somatic mutation of the V617F JAK2 gene in patients of the cardiovascular diseases].

The JAK2 V617F somatic mutation is one of the most frequent markers of CHIP (clonal hematopoiesis of indeterminate potential). CHIP is characterized by the presence of a myeloid cells clone in peripheral blood in the absence of the sufficient reasons to diagnose the hematologic disease. The CHIP is proposed as a potential independent risk factor for vascular pathology. The aim of this study is to identify carriers of JAK2 V617F mutation among patients admitted for planned hospitalization at the Federal Center of Cardiovascular Surgery of Krasnoyarsk. MATERIALS AND METHODS: The study included 930 venous blood samples. JAK2 V617F mutation was detected by using the allele - specific real time polymerase chain reaction. RESULTS: JAK2 V617F mutation was detected in 15 (1.6%) patients, but only two of them had blood cell count that could cause a hematological disease to be suspected. CONCLUSION: The inclusion of the JAK2 V617F mutation detection in the complex of laboratory tests of the cardiovascular patients can facilitate the timely identification of patients with increased thrombotic risk, as well as the timely diagnosis of myeloproliferative diseases.

RESULTS OF EVLT: RADIAL FIBER VERSUS BARE-TIP FIBER WITH CHEMICAL ASSISTANCE.

Purpose of the study - comparison of the effectiveness of various methods of endovenous laser coagulation of the great saphenous vein, searching the optimal scheme for the treatment of chronic venous insufficiency. Retrospective analyze of 318 clinical cases of combined surgical treatment using various methods of thermoablation (bare-tip fiber, radial fiber, simultaneous thermoablation and echosclerobliteration) of a great saphenous vein. By the parameter of the great saphenous vein occlusion, the results in the group of simultaneous thermoablation and echosclerobliteration surpassed the bare-tip thermoablation group results and were comparable with the results of radial thermoablation. The combination of methods of subcutaneous veins occlusion allows to use lower-energy modes of bare-tip endovenous laser coagulation to achieve the effective vein obliteration comparable to the results of radial thermoablation.

A retrospective analysis of adverse effects of an in vivo fluoroquinolone antibiotic enrofloxacin treatment on oocyte quality in the common marmoset.

Here we report a retrospective analysis of negative effects of routine enrofloxacin treatment of recurrent diarrhea on the ovary and the developing oocytes of the common marmoset, a small New World primate. The most deleterious effect on oocytes was observed about two months post treatment suggesting that the enrofloxacin effect is on early growing follicles. Manifestations of toxicity included decreased numbers of growing follicles and recovered culturable oocytes, as well as signs of early atresia of granulosa cells. In addition, increased amounts of holed stroma after treatment strongly suggested increased death of the early growing follicles. Of the oocytes judged to be of adequate quality for culture, maturation rates were not affected but fertilization of in vitro matured MII oocytes and subsequent cleavage rates were severely reduced in the enrofloxacin treated animals. Further, the arrested oocytes, which failed to mature or fertilize, showed obvious meiotic spindle abnormalities.

[The potential of ectoprosthetics in rehabilitation of patients with orbital pathology]. / Vozmozhnosti éktoprotezirovaniia v reabilitatsii patsientov s patologiei orbity.

AIM: rectification of indications and contraindications for ectoprosthetics and exploration of its potential as a modern rehabilitation modality. MATERIAL AND METHODS: Here are summarized the results of 89 cases of facial reconstruction with prosthetic means in trauma patients and those who had had congenital or oncological pathology and in whom other methods of rehabilitation were unable to remedy the defect within the orbital region. RESULTS: The indications and contraindications for facial prosthetics as well as the conditions, under which this type of medical and social rehabilitation can be carried out, were clarified. The authors also explained the process of creating an epiprosthesis and how it should be attached to the orbit to have the maximum effect on the existing defects of both the bone and auxiliary apparatus (including eyelid soft tissues, eyelashes, etc.). Because modern ectoprostheses are made of silicone and held in place with magnets, they provide an optimal cosmetic result, so that, regardless of patient's age, no rough defects can be detected on the affected side at a 2-m distance. CONCLUSION: As the production technology develops, the cosmetic appearance of epiprostheses improves and their use broadens.

Direct actions of androgen, estrogen and anti-Müllerian hormone on primate secondary follicle development in the absence of FSH in vitro.

STUDY QUESTION: What are effects of androgen, estrogen and anti-Müllerian hormone (AMH), independent of FSH action, on the development and function of primate follicles from the preantral to small antral stage in vitro? SUMMARY ANSWER: Androgen and estrogen, but not AMH, promote follicle survival and growth in vitro, in the absence of FSH. However, their growth-promoting effects are limited to the preantral to early antral stage. WHAT IS KNOWN ALREADY: FSH supports primate preantral follicle development in vitro. Androgen and estrogen augment follicle survival and growth in the presence of FSH during culture. STUDY DESIGN SIZE, DURATION: Nonhuman primate model; randomized, control versus treatment groups. Rhesus macaque (n = 6) secondary follicles (n = 24 per animal per treatment group) were cultured for 5 weeks. PARTICIPANTS/MATERIALS, SETTING, METHODS: Follicles were encapsulated in 0.25% (w/v) alginate and cultured individually in modified alpha minimum essential media with (i) FSH (1 ng/ml; control), (ii) no FSH, (iii) no FSH + estradiol (E2; 100 pg/ml)/dihydrotestosterone (DHT; 50 ng/ml) and (iv) no FSH + AMH (50 ng/ml). In a second experiment, follicles were cultured with (i) FSH (1 ng/ml), (ii) no FSH, (iii) no FSH + E2 (1 ng/ml), (iv) no FSH + DHT (50 ng/ml) and (v) no FSH + E2/DHT. Follicle survival, antrum formation and growth pattern were evaluated. Progesterone (P4), E2 and AMH concentrations in culture media were measured. MAIN RESULTS AND THE ROLE OF CHANCE: In the first experiment, FSH deprivation significantly decreased (P < 0.05) follicle survival rates in the no FSH group (16 ± 5%), compared to CTRL (66 ± 9%). E2/DHT (49 ± 5%), but not AMH (27 ± 8%), restored follicle survival rate to the CTRL level. Similarly, antrum formation rates were higher (P < 0.05) in CTRL (56 ± 6%) and E2/DHT groups (54 ± 14%), compared to no FSH (0 ± 0%) and AMH (11 ± 11%) groups. However, follicle growth rate after antrum formation and follicle diameter at week 5 was reduced (P < 0.05) in the E2/DHT group (405 ± 25 µm), compared to CTRL (522 ± 29 µm). Indeed, the proportion of fast-grow follicles at week 5 was higher in CTRL (29% ± 5), compared to E2/DHT group (10 ± 3%). No fast-grow follicles were observed in no FSH and AMH groups. AMH levels at week 3 remained similar in all groups. However, media concentrations of P4 and E2 at week 5 were lower (P < 0.05, undetectable) in no FSH, E2/DHT and AMH groups, compared to CTRL (P4 = 93 ± 10 ng/ml; E2 = 4 ± 1 ng/ml). In the second experiment, FSH depletion diminished follicle survival rate (66 ± 8% in control versus 45 ± 9% in no FSH, P = 0.034). E2 plus DHT (31.5 ± 11%) or DHT alone (69 ± 9%) restored follicle survival rate to the control (FSH) level as expected. Also, E2 plus DHT or DHT alone improved antrum formation rate. However, in the absence of FSH, E2 plus DHT or DHT alone did not support growth, in terms of follicle diameter, or steroid (P4 or E2) production after the antral stage. LIMITATIONS REASONS FOR CAUTION: This study is limited to in vitro effects of E2, DHT and AMH during the interval from the secondary to small antral stage of macaque follicular development. In addition, the primate follicle pool is heterogeneous and differs between animals; therefore, even though only secondary follicles were selected, follicle growth and developmental outcomes might differ from one animal to another. WIDER IMPLICATIONS OF THE FINDINGS: This study provides novel information on the possible actions of estrogen and androgen during early follicular development in primates. Our results suggest that sequential exposure of preantral follicles to local factors, e.g. E2 and DHT, followed by gonadotropin once the follicle reaches the antral stage, may better mimic primate folliculogenesis in vivo. STUDY FUNDING/COMPETING INTEREST(S): Research reported in this publication was supported by the Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Center for Translational Research on Reproduction and Infertility 5P50HD071836, and the NIH Primate Centers Program 8P510D011092. There are no conflicts of interest.

Mechanisms for Strong Anisotropy of In-Plane g-Factors in Hole Based Quantum Point Contacts.

In-plane hole g factors measured in quantum point contacts based on p-type heterostructures strongly depend on the orientation of the magnetic field with respect to the electric current. This effect, first reported a decade ago and confirmed in a number of publications, has remained an open problem. In this work, we present systematic experimental studies to disentangle different mechanisms contributing to the effect and develop the theory which describes it successfully. We show that there is a new mechanism for the anisotropy related to the existence of an additional B_{+}k_{-}^{4}σ_{+} effective Zeeman interaction for holes, which is kinematically different from the standard single Zeeman term B_{-}k_{-}^{2}σ_{+} considered until now.

[The comparative analysis of immunologic techniques of detection of anti-phospholipid antibodies].

The laboratory diagnostic of anti-phospholipid syndrome consists in detection of anti-phospholipid antibodies using technique of enzyme-linked immunosorbent assay namely in detection of anti-cardiolipin antibodies and antibodies to ß2-glycoprotein. In spite of the fact that serological diagnostic plays a key role in diagnosing anti-phospholipid syndrome application of laboratory tests s complicated by their insufficient standardization. The new approach to detection of anti-phospholipid antibodies became application of immune blotting on the basis of polyvinylidenfluoride membrane. As compared with enzyme-linked immunosorbent assay, the advantage of the mentioned technique is in using hydrophobic solid phase for sorption of antigens. The porous structure of polyvinylidenfluoride membrane orientates hydrophilic areas of phospholipids and by that ensures their more dense distribution imitating bi-lipid layer of membranes of living organism. To specify and compare value of different techniques the comparison was implemented concerning the results of measurement of anti-phospholipid antibodies in enzyme-linked immunosorbent assay test-systems of various manufacturers and reagents kits for immune blotting. The collection was assembled including bio-materials from 47 patients with non-cardioembolic ischemic strokes, 20 patients with recurrent thrombosis of deep veins of lower extremities and 50 patients with obstetrics pathology and also 30 healthy donors. In the given serums aKlaIgG, aKlaIgM, aß2glycoprotein I were measured using enzyme-linked immunosorbent assay technique assisted by test-systems of Euroimmun and Orgentes Diagnostica and the samples with the highest titre using immune blotting technique with reagents manufactured by Medipan. On the basis of measurement of anti-phospholipid antibodies by various enzyme-linked immunosorbent assay test-systems the rate of aß2glycoprotein I amounted to 31% in case of Euroimmun reagents kits for enzyme-linked immunosorbent assay, 78% in case of Orgentec Diagnistica test-systems for enzyme-linked immunosorbent assay, aKlaIgG - 2% and 30%, aKlaIgM - 31% and 54% correspondingly. The measurement of anti-phospholipid antibodies using immune blotting technique on Medipan test-systems in bio-samples with the highest titres detected aß2glycoprotein I in all patients, aKlaIgG in 70% and aKlaIgM in 30% of patients. The convergence between three commercial reagents kits varies from 20% to 88%. The standardization of commercial test-systems still to be achieved. The new technique of immune blotting can be appliedjointly with classic techniques ofserological diagnostic of anti-phospholipid syndrome. The absence of algorithms of diagnostic and standardization of different test-systems for detection of anti-phospholipid antibodies prejudices reliability of serological diagnosis of anti-phospholipid syndrome and therefore existence of anti-phospholipid syndrome as a nosologic unit.

Detection of cross-sex chimerism in the common marmoset monkey (Callithrix jacchus) in interphase cells using fluorescence in situ hybridisation probes specific for the marmoset X and Y chromosomes.

Chimerism associated with placental sharing in marmosets has been traditionally analysed using conventional chromosome staining on metaphase spreads or polymerase chain reaction. However, the former technique requires the presence of proliferating cells, whereas the latter may be associated with possible blood cell contamination. Therefore, we aimed to develop a single-cell analysis technique for sexing marmoset cells. We applied fluorescent in situ hybridisation (FISH) to cell nuclei using differentially labelled X and Y chromosome-specific probes. Herein we present the validation of this method in metaphase cells from a marmoset lymphoblastoid cell line, as well as application of the method for evaluation of cross-sex chimerism in interphase blood lymphocytes and haematopoietic bone marrow cells from marmosets of same- and mixed-sex litters. The results show conclusively that haematopoietic cells of bone marrow and leucocytes from blood are cross-sex chimeric when the litter is mixed sex. In addition, single samples of liver and spleen cell suspensions from one individual were tested. Cross-sex chimerism was observed in the spleen but not in liver cells. We conclude that FISH is the method of choice to identify cross-sex chimerism, especially when combined with morphological identification of nuclei of different cell types, which will allow a targeted tissue-specific analysis.

[ENDOSCOPIC MUCOSAL RESECTION AND ENDOSCOPIC DISSECTION IN THE SUBMUCOSAL LAYER FOR EARLY GASTRIC CANCER].

The development of endoscopic techniques of surgery of patients with early gastric cancer (high and moderately differentiated adenocarcinoma) by endoscopic mucosal resection and endoscopic submucosal dissection with regard to their minimally invasion, high efficiency and quality of life after treatment allowed seriously establishing itself and be accepted for use in worldwide.

Body weight-associated differences in ovarian morphology in captive common marmoset (Callithrix jacchus).

In captivity, Callithrix jacchus (common marmoset) is on average heavier than their wild-living counterparts, and has a tendency to produce triplet litters rather than the normal twins. To provide initial basic information about possible weight-related differences among the ovaries, a morphometric study of follicular phase ovaries from 48 young adult marmosets has been carried out. Nearly 90% of these ovaries were found to contain some degree of luteal tissue composed of large and/or small cells. The luteal structures, follicles of all stages, and stroma were subjected to morphometric analysis, and these results were compared with body weight, circulating triglyceride, androstenedione, and total estrogens. Where only large luteal cells were present, the median body weight was the highest (only this group included animals over 500 g) compared with mixed, or only small luteal cells, or absence of luteal cells. Furthermore, in this group plasma triglycerides were significantly higher compared to other groups, suggesting possible role of triglycerides in promoting luteinisation. Plasma androstenedione was also a critical discriminating factor, and was elevated where large luteal cells were present even as a mixture with small cells suggesting the large luteal cells to be the likely major ovarian source of this hormone and its metabolites. Additionally, the ovaries with large luteal cells compared to those containing only small or no luteal cells, had lower primordial follicle reserve associated with high levels of atresia and luteinisation among growing non-ovulatory follicles, indicating an accelerated activation, but at the same time a suboptimal environment for follicular growth.

Critical estradiol dose optimization for oocyte in vitro maturation in the common marmoset.

The aim of the present study was to critically evaluate the effect of different concentrations of estradiol (E2) during IVM of common marmoset (Callithrix jacchus) oocytes from antral follicles. The doses tested were 0, 0.1, 1, or 10 µg/mL E2 (referred to as 0 E2, 0.1 E2, 1 E2, and 10 E2 groups). After a preincubation, the concentration of E2 in IVM drops under oil was approximately 20% of the amount added (0.02; 0.2 and 1.9 µg/mL, respectively) because of absorption into the oil. Oocyte progression to metaphase II was significantly higher in the 0.1 E2 group than that in the absence of E2. With progressively higher doses, the maturation rate tended to decrease suggesting an overdose effect. Furthermore, the total first cleavage rate was significantly higher in the 0.1 E2 group than that in the 0 E2 group and decreased progressively with further increases in E2 concentration, with the 10 E2 group showing the same low rate as without E2. The oocytes which failed to cleave, after maturation in 10 E2, showed obvious signs of overdose with the highest rates of degeneration and abnormal spindle form, and an absence of embryo progression. In contrast to these obvious negative effects on the oocyte, 10 E2 was the only group in which a significant increase in radial cumulus expansion was observed. The concentration 0.1 E2, which is 10 times lower than the most commonly used E2 dose, produced the best results in all oocyte factors evaluated. These results represent the first study for a primate species showing a strong positive effect of E2 on oocyte maturation and embryo development, but only at the optimal concentration, and emphasize the critical limits of the optimal concentration range.