[Clinical and molecular genetic features of cases of isolated hypogonadotropic hypogonadism, associated with defects in GNRHR genes].

Congenital hypogonadotropic hypogonadism (CHH) is a rare disorder characterised by lack of pubertal development and infertility, due to deficient production, secretion or action of gonadotropin-releasing hormone (GnRH). Clinically, there are variants of CHH with hypo-/anosmia (Kalman syndrome) and normosmic hypogonadotropic hypogonadism. Given a  growing list of gene mutations accounting for CHH, the application of next generation sequencing (NGS) comprises an excellent molecular diagnostic approach because it enables the simultaneous evaluation of many genes. Biallelic mutations in GNRHR gene lead to the development of hypogonadotropic hypogonadism with normosmia. In this paper, we describe 16 patients with proven GnRH resistance and estimate the frequency of pathogenic variants in the GNRHR gene in the Russian population.

[Botulinus toxin in complex treatment of myofacial pain syndrome]. / Botulotoksin v kompleksnom lechenii patsientov s miofastsial'nym bolevym sindromom disfunktsii visochno-nizhnecheliustnogo sustava.

The aim of the study was to assess the efficacy of type A Botulinus toxin (BTA) in pain release by TMJ functional pain disorders. The study included 211 patients with TMJ functional pain disorder (20.4% males and 79.6% females; mean age 45.3 years). The patients underwent clinical examination and bioelectric activity assessment of masticatory muscles by electromyography (EMG). EMG specters of 20 healthy volunteers with intact dental arches served as a control. After examination BTA was injected in muscular pain trigger points. All patients had muscular hypertonus, unilateral in 88.6% and bilateral in 11.4%. EMG showed the decrease of masticatory muscle activity on affected side to mean values of 165±20 mkV (30.0%, p<0.05) and on contralateral side to 460±31 mkV (89.6%, p>0.05). BTA injections in tensed muscles released significantly muscle-induced facial pain and improved quality of life. During 6 months follow up myofacial pain disorder relapse was seen in 3 patients. The results allow recommending BTA injection in muscular pain trigger points for treatment of myofacial pain syndrome and prolonged muscle relaxation.

[Similarity of Spectral Profiles with Individual Fluorescence Lifetime of Tryptophan in Proteins of Different Structure].

This work presents the results of the analysis of the fluorescence lifetime of tryptophan in three proteins: human serum albumin, bovine serum albumin and bacterial luciferase, containing 1, 2 and 7 tryptophan residues, respectively. It was shown that for all proteins fluorescence decay can be fitted by three lifetimes: τ1 = 6-7 ns, τ2 = -2,0-2,3 ns and τ3 ≤ 0,1 ns (the native state) and τ1 = 4,4-4,6 ns, τ2 = 1,7-1,8 ns and τ3 ≤ 0,1 ns (the denaturated state). It was found that spectral profiles with individual protein fluorescence lifetime have similar peak wavelength and identical half-width of the spectrum as in the native state (λ(max)τ1 = 342 nm, λ(max)τ2 = 328 nm and λ(max)τ3 = 3i5 nm), and in the denaturated state (λ(max)τ1 = 350 nm, λ(max)τ2 = 343 nm and λ(max)τ3 = 317 nm). In addition, the differences in the steady-state spectra of the studied proteins are caused by the individual ratio of lifetime contributions. The correlation between. lifetime components and a known classification of the tryptophan residues in the structure of proteins, under study was performed within the discrete states model.

Spectral study of fluorone dyes adsorption on chitosan-based polyelectrolyte complexes.

Polyelectrolyte complexes of the chitosan-chondroitin sulfate and chitosan-hyaluronate polycation-polyanion pairs were synthesized and characterized as potential dye adsorbents at different pH levels. Equilibrium isotherm analysis was applied to investigate the efficiency and the mechanism of the adsorption of fluorone dyes (fluorescein, eosin Y, erythrosin B) on the synthesized complexes. The inefficiency of the fluorescein adsorption was proved by two different quantitative spectroscopic methods. The adsorption isotherm for both eosin Y and erythrosin B was adequately described in terms of the Langmuir-Freundlich model. The observed room-temperature phosphorescence of the adsorbed erythrosin B was attributed to the surface inhomogeneity of the synthesized complexes. The revealed variation in the adsorption properties of fluorone dyes was related to the differences in their ionic forms as well as in their polarity and hydrophobicity.

[The two-phase growth medium for sub-culturing of Helicobacter pylori].

A. Pylori is a very undemanding microorganism needing the in support of complex of conditions including particular atmosphere, temperature of culturing and composition of growth medium. The two-phase growth medium is recommended to sub-culturing in Petri dishes with diameter of 90 mm. The growth medium consists of chocolate agar with addition of Schedler broth and enriched with 10% serum of cattle.

[Immunological markers of severity and outcome of traumatic brain injury].

A microscopic immunological study of cerebral spinal fluid (CSF) of 100 patients with traumatic brain injury (TBI) of different severity was carried out in the 1st, 10th and 21st days after trauma. The content of antibodies to basic myelin protein (BMP) and phospholipids (PL) significantly differed from that in the control group (patients with soft tissue head injuries without TBI) and was significantly higher in patients with severe TBI. The level of antibodies to BMP in the CSF was strongly correlated with scores on the Glasgow Coma Scale in the 1st day after TBI and the degree of recovery of neurological status in the 21st day. The level of antibodies to PL was strongly correlated with the severity of cytosis, content of CSF erythrocytes and proteins. The correlation between antibodies to BMP and antibodies to PL was weak. The different diagnostic value of antibodies to BMP and to PL in the most acute period of TBI was found. In conclusion, content of antibodies to BMP may be used as a marker of TBI severity and outcome while the significant elevation of antibodies to PL indicates the severity of vascular complications of trauma.

The role of energy transfer in bioluminescence quenching by xanthene dyes.

The paper reports the experimental results of bioluminescence quenching in the coupled enzyme system NADH:FMN-oxidoreductase-luciferase in the presence of xanthene dyes (fluorescein, eosin Y, erythrosin B) featured by the rate constants of intersystem crossing. From the spectral data and with the help of kinetic model the rate constants of energy transfer from emitter to dye were determined (2.9 x 10(12)-6.5 x 10(13) M(-1) s(-1)) and the emitter lifetime was estimated (1.2-2.7 ns). The calculated rate constants of energy transfer in a series of xanthene molecules are higher than the diffusion-controlled constants. The rate constants of energy transfer correlate with the probability of intersystem crossing of a dye. The combined analysis of emitter fluorescence quenching and acceptor fluorescence enhancement indicates that the significant bioluminescence intensity decrease in the presence of xanthene dyes is determined by both the quenching processes via the energy transfer and the inhibition of bioluminescent reaction.

[The role of antibodies to phospholipids in the pathogenesis of symptomatic epilepsy].

Antibodies to phospholipids (ABPL) were studied in 40 patients, aged from 17 to 65 years, with symptomatic epilepsy. Those with primary or secondary antiphospholipid syndrome and with diffuse diseases of connecting tissues were not included in the study. All patients underwent clinical, neurological, electroencephalographic and immunological examination, the latter including the determination of ABPL level, namely the level of AB to cardiolipin in the blood serum using ELISA. It was shown that the ABPL level was 1,5-2,0 times higher in patients comparing to the controls. This increase was observed in patients with moderate and high frequency of seizures and a history of at least 2 seizures. Moreover, the ABPL level was depended on the etiologic factor and epileptic EEG activity. In conclusion, the results obtained suggest that ABPL not only interact with vessel endothelium but caused the immunologically mediated damage of cortical neurons that plays an important role in the pathogenesis of symptomatic epilepsy.

[Prediction of cerebral disturbances in patients with thermal injury].

In spite of its utter importance, the problem of early prediction of cerebral disturbances (CD) in thermal injury (TI) has not been solved so far due to insufficient knowledge of the pathogenesis of neurological disturbances in burnt patients. Existing prognostic methods are based on the statistical analysis of data, and cannot be considered strictly scientific. The authors of the article offer a new method of prediction of neurological disturbances and the degree of CD in TI, based upon modern conception of the pathogenesis of nervous system lesion in these patients. The method allows prevention of such complications.

[Cerebrovascular disturbances in rheumatoid arthritis].

Rheumatoid arthritis (RA) is a diffuse connective tissue disease and a multi-system disorder with inflammatory process affecting joints in the first place. RA is found in 1 to 3% of population; the first signs of it are usually found in people aged 35 to 50. Neurological pathology in RA is manifested by cervicocranialgia, cervical myelopathy, pathological changes in the upper cervical spine, and cerebral disorders. However, exact mechanisms of the development of central nervous system (CNS) lesions in RA have not been presented. The aim of this study was to clarify the pathophysiological mechanisms and clinical peculiarities of cerebral disturbances in RA. The subjects were 42 female patients, who underwent clinical, neurological, clinicolaboratory, immunological, and clinicophysiological examination. Subjective and objective symptoms were studied; the following syndromes of CNS pathology were distinguished: initial manifestations of cerebral functional insufficiency; disseminated cerebral micro symptoms; focal cerebral lesion. These disorders were accompanied by changes in biochemical parameters which evidenced the presence of connective tissue destruction and immune inflammation. Immunological tests revealed elevation of the level of myelin basic protein antibodies, which correlated with the degree of neurological disturbances and the duration of the disease. The level of myeloperoxidase was elevated, but the degree of this elevation did not depend on the degree of the cerebral disorder and displayed a negative correlation with the duration of the disease. The results of the study demonstrate primary lesion of small vessels in RA--secondary vasculitis followed by demyelinization of CNS white substance. Thus, three forms of cerebrovascular pathology, caused by acute or chronic cerebral vascular insufficiency in RA can be distinguished: initial manifestations of cerebral circulation insufficiency; discirculatory encephalopathy; transient cerebral circulation disturbances and cerebral stroke.

[Clinical, immunological, and electrophysiological matching in Raynaud's syndrome].

Detailed description of Raynaud's syndrome (RS) dates back to the 19th century; nevertheless, this problem is still topical because of high prevalence of the syndrome (4 to 5% of population), and the fact that different specialists have to deal with it. The authors of the article studied clinical, immunological, and electrophysiological peculiarities of 103 patients with RS, both primary and secondary one. The examination included measurement of the level of antibodies to nerve growth factor (NGF) and myeline basic protein (MBP) and electroneuromyography. All the subjects displayed significant elevation of serum titer of MBP and NGF antibodies, and lowered peripheral nerve impulse conduction velocity (ICV). There was a direct correlation between antibody titer and the severity of the disease, and inverse correlation between ICV of sensory nervous fibers and the severity of the disease. Thus, RS is almost always associated with peripheral sensory fiber pathology, whose clinical manifestation consists in demyelinating polyneuropathy of autoimmune origin; the more prominent demyelinization, the higher the degree of disease severity.

[Drug allergy and nervous system disorders].

The article presents data on involvement of the nervous system of patients with medicamentous allergy characterized by allergic lesions of body vessels. Cerebral allergic vasculitis is often masked by other vascular conditions such as the following: atherosclerosis, high blood pressure, rheumatism and vegetovascular dystonia. The use of the reaction of a specific injury of basophilic leukocytes exposed to penicillin, streptomycin may be a diagnostic test in the determination of the damage of the nervous system in patients with medicamentous allergy.

[Monoclonal antibodies CD95 in an acute period of ishemic stroke in arterial hypertension]. / Monoklonal'nye antitela CD95 v ostrom periode ishemicheskogo insul'ta pri arterial'noi gipertonii.

The data on examination of 83 patients in acute period of cerebral stroke caused by arterial hypertension and 30 healthy controls are presented. In all the patients, content of Fas expressed lymphocytes (CD95), indirectly suggesting apoptosis, has been determined. The level of Fas/Apo-1+ lymphocytes was significantly higher comparing to the control group. Cerebral stroke severity positively correlated with Fas/Apo-1+ lymphocytes. The higher was a level of monoclonal antibodies CD95, the poorer was the course and prognosis both for recovering and for life. This correlation argues for a role of Fas/Apo-1+ lymphocytes in the apoptotic process, and the level of their expression reflects that of apoptosis in damaged brain tissue and serves as a sign of immune inflammation in response to acute cerebral disaster.

[Neurosensory deafness: immunological aspects]. / Immunologicheskie aspekty neirosensornoi tugoukhosti pri vpervye vyiavlennom tuberkuleze.

Seven hundred and twelve patients with first diagnosed pulmonary tuberculosis were examined; neurosensory hypoacusis was detected in 12.2% of the cases. The above patients underwent neurological, otolaryngological, and immunological studies. Second- and third-degree hypoacusis was found in 13.7 and 86.3%, respectively. Neurological syndromes were identified. These included vegetovascular dystonia in 50.5% of the patients, multiple cerebral minor symptoms in 45.5%, focal cerebral lesion in 4%/Comparison of the severity of nerve system disease and the level of hypoacusis has indicated that the more severe neurological disorders, neurosensory hypoacusis becomes more aggravated (K = 0.57). Positive skin Mantoux tests in all patients with neurosensory hypoacusis showed their allergization to Mycobacterium tuberculosis. In the examined patients, specific basophilic deformability (SBD) rates in the tuberculin test were 17.5 +/- 1.3% g, which differed significantly (p < 0.001) from those in the controls (5.1 +/- 0.9%). In second-degree neurosensory hypoacusis, SBD rates were significantly higher than those in first-degree one, which suggests that the allergic factor contributes to the development of neurosensory hypoacusis in the presence of tuberculosis. The values of antibodies to myeloperoxidase (1.869 +/- 0.02 optical density units (ODU) and to IgE (416.386 +/- 0.391 I.U./ml) significantly differed from those in the control group (0.08 +/- 0.02 ODU and 96.667 +/- 0.441 I.U./ml, respectively), which is indicative of primary lesions of minute blood vessels as the type of vasculitis and secondary nervous tissue involvement. Thus, allergic, vascular, and immune mechanisms are the main pathophysiological ones of neurosensory hypoacusis in the presence of first diagnosed and untreated tuberculosis.

[The role of autoimmune process in the pathogenesis of hemorrhagic stroke]. / Rol' autoimmunnogo protsessa v patogeneze gemorragicheskogo insul'ta.

Using immunologic study along with clinical and neurological ones, 104 patients in acute stage of hemorrhage stroke (18% of them with subarachnoidal; 27%--with parenchymatous and 55%--with combined hemorrhage) have been examined. A level of antibodies to DNA (anti-DNA) was determined in cerebrospinal fluid (CSF) and blood serum on 1, 10 and 20 days. Control group consisted of 20 persons. Anti-DNA was significantly elevated on day 1 in CSF and blood serum, with the most pronounced increasing in CSF (p < 0.001). The severer was hemorrhage stroke, the higher was anti-DNA level. Furthermore, the poorer was prognosis for recovery and life, the higher elevation was detected for anti-DNA.

[Association of homozygosity for short allele (S) of heavy neurofilament subunit gene with motor neuron disease and oxidative stress development]. / Assotsiatsiia gomozigotnosti po korotkomu alleliu (S) gena tiazhelykh neirofilamentov s bolezn'iu dvigatel'nogo neirona i razvitiem oksidantnogo stressa.

We analyzed distribution of heavy neurofilament (NF-H) gene S/L-polymorphic variants in 51 patients with idiopathic motor neuron disease (MND) vs control group and in relation to superoxide dismutases (SODs) activity and thiobarbituric acid reactive substances (TBARS) level in cerebrospinal fluid (CSF), erythrocytes and blood serum. We found that individuals with homozygosity for NF-H gene short allele (S/S-genotype carriers) in MND group predominate significantly over those in control one (p < 0.001). We revealed significant increase of oxidative markers in CSF and blood serum in MDN patients vs controls (p < 0.05), but not in patients with spondylogenic myelopathy, conforming non-specific role of oxidative stress in MND pathogenesis. There were no differences between TBARS level in CSF and serum in relation to the rate of MND progression, suggesting that oxidative stress does not influence the MND course. We showed normal SOD-1 activity in erythrocytes and CSF of MND patients that argued for the absence of these antioxidant enzymes deficiency in MND without SOD-1 gene mutations. We found significant association between homozygosity for short allele (S) and increased TBARS level in CSF (p < 0.02). These findings specify the role of NF-H with lower molecular weight in MND pathogenesis and make expedient antioxidants administration to MND patients homozygous for S-allele of NF-H gene.

[Blood infrared spectrometry in progressive myodystrophy]. / Infrakrasnaia spektrometriia krovi pri progressiruiushchikh myshechnykh distrofiiakh.

Twenty four patients with various progressive myodistrophy (PMD) types have been examined. Initial and follow-up blood counts, using infrared spectrometry, were carried out. All the patients had phospholipid dysbolism. Its level positively correlated with myodistrophy severity. Phospholipid level decrease was associated with membrane permeability increase. The authors consider phospholipid dysfunction as one of the key PMD pathogenetic mechanisms. Blood infrared spectrometry is recommended for PMD severity determination, in diagnosis and therapy which is pathogenetically substantiated.

Effects of potassium halides on bacterial bioluminescence.

The effects of potassium halides KCl, KBr and KI on NADH:FMN-oxidoreductase-luciferase bioluminescent coupled enzyme system were studied. The influence of salt additions on bioluminescence intensity and bioluminescence light yield was investigated. The inhibition and activation parameters of the salts were calculated using their dependencies on concentration of the salts. The correlation between the inhibition of bioluminescence intensity and the halide mass was demonstrated: the inhibiting ability of the salts increases with the increase of atomic weight of the anions. The inhibition parameters increase and the activation parameters decrease, accordingly.