Microbiological synthesis of stereoisomeric 7(α/ß)-hydroxytestololactones and 7(α/ß)-hydroxytestolactones.

Microbiological synthesis of 7α- and 7ß-hydroxy derivatives of testololactone and testolactone was developed based on bioconversion of dehydroepiandrosterone (DHEA) by fungus of Isaria fumosorosea VKM F-881 with subsequent modification of the obtained stereoisomers by actinobacteria. The first stage included obtaining of the stereoisomers of 3ß,7(α/ß)-dihydroxy-17a-oxa-D-homo-androst-5-en-17-ones in the preparative amounts. Then the conversion of 7-hydroxylated D-lactones obtained by selected actinobacteria of Nocardioides simplex VKM Ac-2033D, Saccharopolyspora hirsuta VKM Ac-666, and Streptomyces parvulus MTOC Ac-21v was studied. Under the transformation of 3ß,7α-dihydroxy-17a-oxa-D-homo-androst-5-en-17-one and its corresponding 7ß-stereoisomer by N. simplex VKM Ac-2033D and S. hirsuta VKM Ac-666 the 7α- and 7ß-hydroxy-17a-oxa-D-homo-androst-4-ene-3,17-dione (7α- and 7ß-hydroxytestololactone), 7α- and 7ß-hydroxy-17a-oxa-D-homo-androsta-1,4-diene-3,17-dione (7α- and 7ß-hydroxytestolactone) were obtained with molar yields in a range of 60.3-90.9 mol%. The crystalline products of 7α-hydroxytestololactone, 7α-hydroxytestolactone, and their corresponding 7ß-hydroxy stereoisomers were isolated, and their structures were confirmed by mass spectrometry and 1H-NMR spectroscopy analyses. The strain of Str. parvulus MTOC Ac-21v transformed 3ß,7(α/ß)-dihydroxy-17a-oxa-D-homo-androst-5-en-17-ones into the corresponding 3-keto-4-ene analogs and did not show 3-ketosteroid 1(2)-dehydrogenase activity. The activity of actinobacteria towards steroid D-lactones was hitherto unreported.The results contribute to the knowledge of metabolic versatility of actinobacteria capable of transforming steroid substrates and may be applied in the synthesis of potential aromatase inhibitors.

[Use of somatostatin analogues in the pancreonecrosis therapy (kliniko-experimental research)]. / Osobennosti primeneniya sinteticheskogo analoga somatostatina v terapii pankreonekroza (kliniko-eksperimental'noe issledovanie).

The article is devoted to the study and optimization of therapeutic strategy in pancreatic necrosis. We studied the effectiveness of clinical application of human somatostatin analog - octreotide in different dosages. Submitted investigationconsists of experimental and clinical parts. MATERIAL AND METHODS: Experimentalmodel of acute destructive pancreatitis onrats of Wistar line with weight of 300±30 gwas reproduced.There were 5 animal groups (one control group and four experimental groups). Experimental animals received oktreotidin different doses. In 2 days there were estimated the blood biochemistryand histopathologic features of liver and pancreas of operated animals. In clinical part there were formed two groups of patients out of selected case histories by a principle of similarity of complex therapy differing in oktreotid doses, namely: 1) pancreatonecrosis patients estimated of 3 to 8 points by Ranson scale in whose treatment regimen oktreotid 300 mkg/day (n=70) dose was included; 2) pancreatonecrosispatients estimated of 3 to 8 points by Ransonin whose treatment regimenoktreoid1200 mkg/day (n=38) was included. There were compared indices of lethal outcomes, average bed-days, and dynamics of laboratory findings, number and volume of surgical procedures. RESULTS AND CONCLUSION: Dose-related effect of a synthetic analogue of somatostatin, acetateoktreotid, was proved. Introduction in complex therapy of pancreatonecrosisthe highest possible authorized doses of oktreotid (1200 mkg/day) is conducive to more favorable course of illness, dynamic reduction of amilazemia, decrease of lethal outcomes and total bed-day, and also, reduction of initial and repeated operative procedures.

[11beta-Hydroxylation of 6alpha-Fluoro-16alpha-Methyl-Deoxycorticosterone 21-Acetate by filamentous fungi].

Selected filamentous fungi--98 strains of 31 genera--were screened for the ability to catalyze 11beta-hydroxylation of 6alpha-fluoro-16alpha-methyl-deoxycorticosterone 21-acetate (FM-DCA). It was established that representatives of the genera Gongronella, Scopulariopsis, Epicoccum, and Curvularia have the ability to activate 11beta-hydroxylase steroids. The strains of Curvularia lunata VKM F-644 and Gongronella butleri VKM F-1033 expressed maximal activity and formed 6lpha-fluoro-16alpha-methyl-corticosterone as a major bioconversion product from FM-DCA. The structures of the major products and intermediates of the bioconversion were confirmed by TLC, H PLC, MS and 1H NMR analyses. Different pathways of 6alpha-fluoro-16alpha-methyl-corticosterone formation by C. lunata and G. butleri strains were proposed based on intermediate identification. The constitutive character and membrane-binding localization were evidence of a 11beta-hydroxylating system in G. butleri, while an inducible character and microsomal localization was confirmed for 11beta-hydroxylase of C. lunata. Under optimized conditions, the molar yield of 6alpha-fluoro-16alpha-methyl-corticosterone reached 65% at a FM-DCA substrate loading of 6 g/L.

Streptomyces tsukubensis VKM Aс-2618D-an Effective Producer of Tacrolimus.

The Streptomyces sp. VKM Ac-2618D strain has been identified, and its morphological and physiological features have been studied in relation to the production of the immunosuppressant tacrolimus. The phenotypic variability of the strain was analyzed, and a dissociant with a high level of tacrolimus production was selected. Based on a comprehensive study of morphological, physiological, and chemotaxonomic properties and on phylogenetic analysis, the strain was named Streptomyces tsukubensis VKM Ac-2618D. The strain genome contains the full version of the tacrolimus biosynthetic gene cluster. The advantages of fed-batch cultivation mode for tacrolimus biosynthesis are shown. The results broaden the understanding of the characteristics of polyketide biosynthesis and can be used in the development of technology for tacrolimus production.

[Bioconversion of C19- and C21-steroids with parent and mutant strains of Curvularia lunata].

Regio- and stereospecificity of microbial hydroxylation was studied at the transformation of 3-keto-4-ene steroids of androstane and pregnane series by the filamentous fungus of Curvularia lunata VKMF-644. The products of the transformations were isolated by column chromatography and identified using HPLC, mass-spectrometry (MS) and proton nuclear magnetic resonance (1H NMR) analyses. Androst-4-ene-3,17-dione (AD) and its 1(2)-dehydro- and 9alpha-hydroxylated (9-OH-AD) derivatives were hydroxylated by the fungus mainly in position 14alpha, while 6alpha-, 6beta- and 7alpha-hydroxylated products were revealed in minor amounts. At the transformation of C21-steroids (cortexolone and its acetylated derivatives) the presence of 17-acetyl group was shown to facilitate further selectivity of 11beta-hydroxylation. Original procedures for protoplasts obtaining, mutagenesis and mutant strain selection have been developed. A stable mutant (M4) of C. lunata with high 11beta-hydroxylase activity towards 21-acetate and 17alpha,21-diacetate of cortexolone was obtained. Yield of 11beta-hydroxylated products reached about 90% at the transformation of 17alpha, 21-diacetate of cortexolone using mutant strain M4.

[Efficacy of rehabilitation of patients with chronic primary low back pain at the spa Klyuchi using balneopelotherapy and transcranial electrostimulation].

Low back pain caused by vertebrogenic pathology is encountered in 60-80% of adult population. We studied efficacy of balneopeloid factors of Klyuchi health resort (Perm Region) and transcranial electrostimulation (TCE) in 60 patients with primary chronic low back pain. The patients were divided into two groups. Thirty patients of group 1 received balneopelotherapy (sulfide mud), Thirty patients of group 2 were given the same balneopelotherapy plus TCE. Pain relieved and psychovegetative status improved in both groups but the effect was more pronounced in group 2.

[Quantitative determination of serum serotonin in the diagnosis of tension headache].

A comprehensive psychometric testing and measurement of the serum levels of serotonin by enzyme immunoassay were made before and after treatment with the antidepressant Prodep in 76 patients with chronic tension headache. The patients generally had cephalgia and moderate depression, which were associated with the lower blood concentration of serotonin. A course of antidepressant therapy with Prodep ameliorated headache and depression in the presence of higher serum serotonin levels. Serum serotonin may be regarded as a marker of the severity of pain and depression in patients with chronic tension headache and be an objective indicator of the efficiency of antidepressant therapy.

[Method for determining leukocytic lipid-releasing capacity in the diagnosis of mechanisms responsible for atherogenesis in patients with coronary heart disease and ischemic stroke].

The authors have studied the mechanism responsible for neutrophilic accumulation and release of protein-lipid complexes in patients with atherosclerosis and the clinical value of this reaction. They present the results of experimental studies and clinical observations in the groups of patients with ischemic stroke, coronary heart disease, bronchial asthma, chronic obstructive pulmonary disease, and rheumatoid arthritis. Whether the process of neutrophilic and release of protein-lipid can be in vitro modulated in a 3-day culture has been demonstrated. The high diagnostic value of the method for determining the lipid-releasing capacity of leukocytes is shown. Some proteins involving in the formation of pathogenic protein-lipid complexes are identified. There is evidence for the use of new differential criteria for visceral diseases originating from atherosclerosis.

[Serum serotonin in patients with tension headaches].

Tension headache (TH) is one of the most frequent types of idiopathic headaches. The leading role in its pathogenesis is played by depression and dysmetabolism of the neurotransmitter serotonin. The subjects were 100 patients with TH. The examination included study of headache intensity, complex psychometric testing, and immune-enzyme measurement of serotonin serum level, performed before and after treatment with the anti-depressant prodep. All the patients had moderate pain syndrome, depression, and moderate or severe anxiety, which demonstrated negative correlation with serotonin serum level. In particular, the intensity of episodic THs (n = 24) was 52 mm visual analogue scale, reactive anxiety level was as high as 51.08 +/- 4.2, the level of depression was moderate (12.9 Beck scale); serotonin blood level showed a tendency to fall (205.72 +/- 6.74 ng/ml). In 76 patients, suffering from chronic THs with a cephalgia intensity of 62 mm visual analogue scale, high indexes of reactive and personal anxiety (46.81 -/+ 2.68 and 54.2 +/- 3.64, respectively), and high depression level were associated with a prominent decrease of serotonin blood concentration (119.38 +/- 9.42 ng/ml). A course of treatment with prodep led to significant decrease of headache intensity and improved depression, while an increase of serum serotonin level was observed. Thus, serotonin serum level may be used as a marker of pain intensity and the level of depression, and an objective indicator of anti-depressive therapy.

[Arterial hypotension and the initial manifestations of cerebral circulatory failure]. / Arterial'naia gipotoniia i nachal'nye proiavleniia nedostatochnosti mozgovogo krovoobrashcheniia.

A study was made of the health status of 20 persons examined for the first time 10 years before for a long-term and steady decrease of arterial pressure classified at that period as vegetodystonia of the neurotic character. 71 healthy test subjects served as a reference group. In addition to the clinical examination, a specialized questionnaire was used to examine the status of the vegetative nervous system; the actual mental status and personality traits were investigated by the MIL test modified by F. B. Berezin and M. P. Miroshnikov; the level of reactive and personality anxiousness were estimated with the aid of Spilberger's scale; the life events and changes were assessed as well. It has been revealed that vegetodystonia recorded in the disease history produces no effect on the frequency and times of the appearance of the initial manifestations of cerebral circulatory failure (IMCCF). At the same time the intensity of vegetative disorders in the structure of the clinical manifestations of the IMCCF was higher in persons, in whose disease histories vegetodystonia was recorded.

[The cerebral mechanisms of the autonomic disorders in vibration disease]. / Tserebral'nye mekhanizmy vegetativnykh narushenii pri vibratsionnoi bolezni.

The status of the nonspecific brain systems was analyzed in 72 miners. Of these, 41, were in the premorbid stage and 31 were with the initial manifestations of vibration disease that occurred as a result of exposure to local vibration. The miners demonstrated dysfunction of the limbic-reticular complex in the form of concomitant vegetative, psychoemotional and nonspecific activation shifts. The syndrome of vegetative dystonia characteristic of the early (preclinical and manifest) nonspecific manifestations of vibration disease was a clinical reflection of cerebral dysfunction.

[Role of cerebral mechanisms in the development of arterial hypotension]. / Rol' tserebral'nykh mekhanizmov v formirovanii arterial'noi gipotenzii.

On the basis of clinical and experimental studies of 83 patients with primary arterial hypotension, the authors present characteristic features of its clinical manifestations related to genetic predisposition and the effect of exogenous factors. Emotional, vegetative, hemodynamic and cerebral disorders associated with the clinical form of disease, namely with the presence or absence of syncopes in the picture of disease are analyzed. Characteristics of psychovegetative disturbances and also of the EEG pattern in various emotional disorders are offered. The results obtained seem to suggest the involvement of cerebral systems in the disruption of hemodynamic mechanisms in arterial hypotension.

[Autonomic disorders in the clinical picture of pathogenetic variants of sclerocystic ovaries]. / Vegetativnye narusheniia v klinike patogeneticheskikh variantov sklerokistoza iaichnikov.

On the basis of examination of the vegetative nervous system in 177 patients with sclerocystic ovaries (SCO) the authors conclude that the clinical picture of the disease is constantly characterized by a clear-cut extragenital symptomocomplex of vegeto-vascular dystonia (a psychovegetative syndrome). The severity and time of its development impart a certain peculiarity to every pathogenetic variant of SCO which may be used for the diagnosis of clinical forms of the disease. SCO are characterized in neuro-psychosomatic terms. It is recommended that psycho- and vegeto-trophic drugs be included into the therapeutic complex.

[Disorders of thyroid function as a consequence of severe craniocerebral injury]. / O narusheniiakh funktsii shchitovidnoi zhelezy vsledstvie tiazheloi cherepno-mozgovoi travmy.

In 32 patients with acute severe head trauma (SHT) the venous plasma levels of thyroid and thyrotrophic (TTH) hormones were studied and the functional state of the hypothalamus-hypophysis-thyroid system assessed. TTH hypersecretion and steady thyroid hypofunction are regular responses of the thyroid endocrine complex to SHT. Pathogenic mechanisms of the adenohypophyseal and thyroid dysfunctions in acute SHT are discussed and the need in their correction suggested.

[Electroaerosol therapy as a pathogenetic method in the complex treatment of arterial hypotension]. / Elektroaérozol'terapiia kak patogeneticheskii metod v kompleksnom lechenii arterial'noi gipotenzii.

On the basis of clinico-experimental examination of 83 patients with primary arterial hypotension (AH) in the process of treatment with electroaerosols, the authors characterize the adjustment of the cardiovascular system parameters, emotional manifestations and cerebral disturbances. The normalization of the vegetative, emotional and other pathological manifestations in the course of the treatment is associated with the use of positively charged electroaerosols. It is concluded that positively charged aerosols can be used in the multiple modality treatment of patients with AH in various types of the disease course.

[A psychoautonomic syndrome in the residual period of a modern "mild" military craniocerebral trauma]. / Psikhovegetativnyi sindrom v rezidual'nom periode sovremennoi "legkoi" boevoi cherepno-mozgovoi travmy.

As many as 18 patients with a history of military service in Afghanistan, who suffered (7.2 years before on the average) commotion due to the explosion wave were examined. The vegetative status was studied by different methods including cardiointervalography, as was the patients' emotional and personality sphere. The data obtained were compared to those obtained in three control groups (practically healthy subjects; those who suffered a mild home craniocerebral injury; healthy subjects who fought in Afghanistan and were not victims to craniocerebral injury). The main group patients demonstrated noticeable changes in the vegetative tone, vegetative reactivity and vegetative supply to physical activity as well as a high level of anxiety, which forms vegetodystonia. Attempts were made to correct the psychovegetative syndrome by central electroanalgesia in the tranquilization mode (9-10 daily sessions). The lowering of the tension of regulatory body systems functioning and the reduction of anxiety and depression tendencies were recorded.

[Autonomic disorders in the formation of the clinical manifestations of the initial forms of cerebrovascular lesions]. / Vegetativnye narusheniia v formirovanii klinicheskikh proiavlenii nachal'nykh form sosudistykh porazhenii golovnogo mozga.

Among 103 persons aged 20 to 57 years, related occupationally to emotional stress, 48 persons were found to have vegetative dystonia, 28 initial manifestations of cerebral circulation failure, and 27 persons appeared healthy. The determination of the character and intensity of vegetative manifestations has shown that in the formation of vegetative dystonia, of paramount importance are cardiovascular and gastrointestinal disorders. Vegetative manifestations associated with initial manifestations of cerebral circulation failure include, in addition, respiratory disturbances and impairment of sleep function, which may reflect more deep changes in the activity of the regulatory apparatus in dyshemias of organic nature. A relationship was discovered between vegetative lesions and emotional disorders, particularly the level of anxiety, as was the contingency of vegetative dysfunction and decreased emotional stress tolerance.

[Multiple sclerosis in childhood]. / Rasseiannyi skleroz v detskom vozraste.

The authors discuss the problems of the age-associated range of multiple sclerosis, characteristics of its clinical manifestations and course in childhood. Provide two cases of multiple sclerosis that had set in 11-year- and 14-year-old children. The first case was marked by a polysymptomatic debut with retrobulbar neuritis of the optical nerves, astereognosis and atetoid hyperkinesis in the hand. The second case was characterized by remitting oculomotor, vestibular, pyramidal disorders associated with facial neuropathy.

[Changes in cerebral blood circulation and their correction in the interparoxysmal period of epilepsy]. / Izmeneniia tserebral'noi gemotsirkuliatsii i ikh korrektsiia v mezhparoksizmal'nom periode épilepsii.

The linear cerebral blood flow (LCBF) was explored in 75 patients with epilepsy according to 131I-albumin transition. LCBF acceleration on the side of the focus, during which the attacks occur rarely and the background EEG remains unchanged is regarded as the most adequate LCBF. The grave course of epilepsy is associated with a decrease of the LCBF on the side of the focus. The pattern of LCBF alterations depends as well on the form of epilepsy. Subclinical activation of the epileptic focus by corasole or hyperventilation gives rise to an increase of the LCBF on its side in patients with generalized secondary epilepsy, to a lowering in temporal epilepsy, and the lack of dynamics in the precentral disease pattern. Correction of the hemodynamic disorders can be attained with the aid of combining phenobarbital and caffeine or papaverine. The pathogenetic importance of the hemodynamic disorders is discussed in connection with functional hypoxia of neurons.

[Epidemiology and clinical polymorphism of hereditary ataxia]. / Epidemiologiia i klinicheskii polimorfizm nasledstvennykh ataksii.

The paper is concerned with a clinico-genetical analysis of 66 patients with hereditary ataxia from 44 families. The authors describe typical and atypical variants of separate clinical forms, underline the intra-and interfamilial polymorphism of the clinical picture. The study contains a detailed description of intellectual and extraneuronal changes, a characteristics of the EEG, EMG and REG changes.