[The procedural aspects of postmortem defense of non-material goods of patient].

The protection of privacy of patient that is applying for medical care, diagnostics and treatment is one of the main pillars of implementation of medical activities. However, despite sufficient volume of legal regulation of lifetime respect of privacy of examination, observation and facts of medical intervention, in practice occur situations when privacy ceases to be such after death of the patient. The article is based on results of content-analysis of normative legal acts (n = 11), scientific publications (n = 52), cases of judicial practice (n = 8). The problem of non-observance of privacy becomes especially actual after death of person whose name is widely known. Having no possibility to impact on causes and modes of obtaining and propagating such specific information, the deceased, one's illness, stages of treatment, struggle for life, resources spent, clinics and specialists involved become object of close attention of various subjects (mas media, Internet communities) that use obtained information for speculative purposes at the expense of fixation of inhumane public replicas and increase of number of browsing. Despite prohibition provided for by law of dissemination of information constituting privacy of applying for medical care, after death of patient and application of measures of legal liability for fact of such a disclosure, information leak occurs regularly and information that got into common access frequently discredit good name of the deceased or develop other problems that are subjects to legal protection, for family members and legal successors. Yet, circumstances of transfer of information of limited access by medical organization (data leakage) or by intent of medical worker are difficult to be proved. Therefore, in judicial practice there are practically no such cases. At that, after death of famous people, juristic community regularly faces problems of protecting personal rights and non-material values from the side of relatives, legal successors and other interested persons. On the basis of formal logical method and system analysis method of disputable situations, possible perspectives of applying for judicial protection, procedural characteristics of procedure of applying for protection and possible composition of participants are presented.

Human decidual gamma/delta T cells possess unique effector and TCR repertoire profiles during pregnancy.

Human γδ T cells are enriched at the maternal-fetal interface (MFI, decidua basalis) showing a highly differentiated phenotype. However, their functional potential is not well-known and it is not clear whether this decidua-enrichment is associated with specific γδ T cell receptors (TCR) as is observed in mice. Here we addressed these open questions by investigating decidual γδ T cells during early and late gestation, in comparison with paired blood samples, with flow cytometry (cytotoxic mediators, cytokines) and TCR high-throughput sequencing. While decidual γδ T cells expressed less perforin than their counterparts in the blood, they expressed significant more granulysin during early pregnancy. Strikingly, this high granulysin expression was limited to early pregnancy, as it was reduced at term pregnancy. In contrast to this granulysin expression pattern, decidual γδ T cells produced reduced levels of IFNγ and TNFα (compared to paired blood) in early pregnancy that then increased by term pregnancy. TCR repertoire analysis indicated that human decidual γδ T cells are not generated early in life as in the mouse. Despite this, a specific enrichment of the Vγ2 chain in the decidua in early pregnancy was observed that disappeared later onwards, reflecting dynamic changes in the decidual γδ TCR repertoire during human gestation. In conclusion, our data indicate that decidual γδ T cells express a specific and dynamic pattern of cytotoxic mediators, Th1 cytokines and TCR repertoire suggesting an important role for these unconventional T cells in assuring a healthy pregnancy in human.

The dose-, LET-, and gene-dependent patterns of DNA changes underlying the point mutations in spermatozoa of Drosophila melanogaster. I. Autosomal gene black.

Sequence analysis of 7 spontaneous, 27 γ-ray- and 20 neutron/neutron+γ-ray-induced black (b) point mutants was carried out. All these mutants were isolated as non-mosaic transmissible recessive visibles in the progeny of irradiated males from the wild-type high-inbred laboratory D32 strain of Drosophila melanogaster. Among spontaneous mutants, there were two (28.5 %) mutants with copia insertion in intron 1 and exon 2, three (42.8 %) with replacement of b+D32 paternal sequence with maternal b1 sequence (gene conversion), one (14.3 %) with 142-bp-long insertion in exon 2, and one (14.3 %) with a short deletion and two single-base substitutions in exon 3. Among γ-ray-induced mutants, there were 1 (3.7 %) with copia insertion in intron 2, 6 (22.2 %) with gene conversion, and the remaining 20 (74.1 %) mutants had 37 different small-scale DNA changes. There were 20 (54.1 %) single- or double-base substitutions, 7 (18.9 %) frameshifts (indels), 9 (24.3 %) extended deletions or insertions, and 1(2.7 %) mutant with a short insertion instead of a short deletion. Remarkably, clusters of independent small-scale changes inside the gene or within one DNA helical turn were recovered. The spectrum of DNA changes in 20 neutron/ neutron+γ-ray-induced mutants was drastically different from that induced by γ-rays in that 18 (90.0 %) mutants had the b1sequence. In addition, 2 (10.0 %) with gene conversion had 600- or 19-bp-long deletion in exon 3 and 1 (5.0 %) mutant with a short insertion instead of a short deletion. Analysis of all 27 mutants with gene conversion events shows that 20 (74.1 %) had full b1 sequence whereas 7 others (25.9 %) contained a partial b1 sequence. These data are the first experimental evidence for gene conversion in the early stages of animal embryogenesis in the first diploid cleavage nucleus after male and female pronuclei have united. The gene conversion, frameshifts (indels), and deletions between short repeats were considered as products of a relevant DNA repair pathways described in the literature. As the first step, the gametic doubling doses for phenotypic black point mutations and for intragenic base substitution mutations in mature sperm cells irradiated by 40 Gy of γ-rays were estimated as 5.8 and 1.2 Gy, respectively, showing that doubling dose for mutations at the molecular level is about 5 times lower than that at the phenotypic level.

Effect of Transplantation of Neural Stem and Progenitor Cells on Memory in Animals with Alzheimer's Type Neurodegeneration.

The effects of systemic and intracerebral transplantation of human fetal neural stem and progenitor cells were studied on the model of olfactory bulbectomy in mice with developing signs of sporadic Alzheimer's disease. It was found that transplantation of these cells at certain stages of disease development contributed to improvement of spatial memory and preservation of hippocampal neurons in these animals.

[Stimulating effect of water-soluble fraction from brown algae Fucus vesiculosus extract on cell proliferation in tissue culture of young and old rats.]

The influence of water-soluble fraction F-55 of brown algae extract Fucus vesiculosus on the development of organotypic tissue culture of different genesis, originating from three germ leaves in young and old rats was studied. The preparation F-55 has a stimulating effect on the processes of cell proliferation in the tissues of ecto-, ento- and mesodermal genesis (cerebral cortex, myocardium, spleen and liver), by regulating the processes of cell proliferation and apoptosis. The revealed strengthening of regeneration processes in explants from both young and old rats under the influence of F-55 preparation in the range of ultra-low concentrations of 10-100 ng/ml can serve as a basis for the creation of new medicinal cytoprotective substances that allow to enhance cell regeneration in various tissues in pathological conditions, including those associated with age.

[Mental disorders in patients with acromegaly]. / Psikhicheskie rasstroistva u bol'nykh akromegaliei.

AIM: To study mental disorders in acromegaly due to somatotropinoma. MATERIAL AND METHODS: The study included 115 consecutively recruited patients with acromegaly (95 female and 20 male, aged from 21 to 78 years). Acromegaly was diagnosed by endocrinologists according to current guidelines based on clinical, laboratory data and brain MRI. All patients underwent a clinical psychiatric interview based on ICD-10 criteria. The Mini-Mental State Examination scale and Hypomania-Checklist (HCL-32) were used. RESULTS: Mental disorders were diagnosed in 79.1% of patients. Organic spectrum disorders were found in 46.1%, bipolar spectrum disorders in 35.7%, schizophrenia spectrum disorders in 4.3%. The patients with bipolar spectrum disorders had significantly lower serum insulin-like growth factor 1 (IGF-1) levels compared to patients with organic spectrum disorders (p=0.01). The presence of organic spectrum disorders was associated with older age and number of somatic comorbidities (р=0.0001 and 0.001). CONCLUSION: The prevalence of bipolar, organic and schizophrenia spectrum disorders in patients with acromegaly exceeds that in the general population. Significantly lower IGF-1 levels in acromegalic patients with bipolar disorders, compared to those with organic disorders, can have some implications to their pathogenesis.

Effect of americium-241 on luminous bacteria. Role of peroxides.

The effect of americium-241 ((241)Am), an alpha-emitting radionuclide of high specific activity, on luminous bacteria Photobacterium phosphoreum was studied. Traces of (241)Am in nutrient media (0.16-6.67 kBq/L) suppressed the growth of bacteria, but enhanced luminescence intensity and quantum yield at room temperature. Lower temperature (4 °C) increased the time of bacterial luminescence and revealed a stage of bioluminescence inhibition after 150 h of bioluminescence registration start. The role of conditions of exposure the bacterial cells to the (241)Am is discussed. The effect of (241)Am on luminous bacteria was attributed to peroxide compounds generated in water solutions as secondary products of radioactive decay. Increase of peroxide concentration in (241)Am solutions was demonstrated; and the similarity of (241)Am and hydrogen peroxide effects on bacterial luminescence was revealed. The study provides a scientific basis for elaboration of bioluminescence-based assay to monitor radiotoxicity of alpha-emitting radionuclides in aquatic solutions.

Chlorsulfuron resistant transgenic tobacco as a tool for broomrape control.

Broomrape (Orobanche ramosa L.) is the most important parasitic plant that infests tobacco (Nicotiana tabacum L.). Chemical treatment of the soil is not effective and crop rotation is not acceptable to solve this problem because of the long viability period of Orobanche seeds in the soil. Application of systemic herbicides in the field with herbicide resistant tobacco could be a successful tool for broomrape control. Several tobacco cultivars were transformed with a mutant ahas3R gene for resistance to the herbicide chlorsulfuron (Glean, DuPont). Transformed plants were selfed and the segregation of resistance was followed in the next generation. The efficiency of the herbicide was demonstrated in greenhouse and field trials. An Orobanche/tobacco growth system was used in order to prove the lethal effect of the herbicide to the attached broomrape plants.

Oxidative stress in blood in cases of untreated refractory anaemia.

PURPOSE: To examine some features of free radical processes in the blood of patients with untreated refractory anaemia (RA) and to interpret their pathogenetic role. MATERIALS AND METHODS: Products of the lipid peroxidation (malondialdehyde-MDA) in whole blood, some antioxidant systems - superoxide dismutase (SOD) in erythrocytes, catalase (CTS) activity and the concentration of sulfhydryl groups (SHG) in whole blood, as well as the spontaneous and stimulated chemiluminescent activity of polymorphonuclear leucocytes (PMNL), reflecting the production of oxygen free radicals, were studied in the blood of 21 patients suffering from untreated RA (study group) and in 45 healthy individuals (control group). RESULTS: Increased content of MDA (p<0.001), lowered level of CTS activity (p<0.001) and concentration of SHG (p<0.05), increased SOD activity (p<0.05) and an increase in the spontaneous PMNL oxidative activity (p<0.05) were found in the study group compared with the control group. CONCLUSION: Strong oxidative stress was recorded in the blood of patients with untreated RA. An assumption was made that the initial iron overload helps the initiation of free radical oxidative processes in blood, accompanied with spontaneous activation of PMNL. Erythrocyte membranes probably are the main target for oxidative attack.

Changes in phagocyte activity in patients with ischaemic stroke.

The activity of peripheral phagocytes to generate reactive oxygen species (ROS) was studied in healthy individuals and patients with ischaemic stroke. The aim was to clarify the relationship between phagocyte activity, the time elapsed after the onset of disease and stroke severity. The total and extracellular production of ROS were evaluated by luminol chemiluminescence. Simultaneously the plasma oxidant activity was determined. When stimulated by opsonized zymosan, phagocytes in patients with stroke (regardless of its severity) showed fast activation. The total ROS generation increased over time in all stroke cases studied. However, the extracellular ROS generation was found to be greater in patients with severe stroke than in those with mild neurological deficiency. When stimulated by formyl-methionyl-leucyl-phenylalanine, the total oxidative phagocyte capacity (regardless of stroke severity) increased over time, but there was no change in the amount of extracellularly generated ROS. In patients with stroke the oxidant activity of plasma was enhanced. We conclude that circulating phagocytes in patients with ischaemic stroke are primed for enhanced ROS production by opsonin receptor-mediated stimulation and for increased secretion of myeloperoxidase by opsonin receptor-independent stimulation. The enhanced extracellular generation of ROS through opsonin receptor-dependent stimulation may be considered an oxidative stress biomarker in cerebral ischaemia.

Functional states of polymorphonuclear leukocytes determined by chemiluminescent kinetic analysis.

During the respiratory burst, upon stimulation with both soluble and particulate matter, polymorphonuclear leukocytes (PMN) generate reactive oxygen species (ROS) and emit chemiluminescence (CL) as a result of metabolic activation. The measurement of CL has been demonstrated to be a useful tool for in vitro assessment of the opsonophagocytic function of PMN. Using component analysis of CL kinetics, we characterized the functional state of PMN by three parameters of the respiratory burst: capacity, effectiveness and velocity (CEV space). The possibility of delimiting eight different functional states of PMN is discussed. The CL kinetics shown by blood PMN in different functional states was analysed, and revealed six out of eight functional states. We conclude that CEV-estimated functional states of PMN are relative, depending on both PMN readiness to generate ROS and conditions of the CL test.

Model components of luminol chemiluminescence generated by PMNL.

The production of activated oxygen species (AOS) by neutrophils (PMNL) is thought to play a key role in the host defence against invading microorganisms. However, the oxygen metabolites are toxic not only to the invading bacteria but also to the surrounding tissue. The oxidative metabolites production can be evaluated by means of chemiluminescent methods. In this study, the possibility of a new analytical approach for quantitative assessment of chemiluminescent kinetics (AOS generation) of isolated PMNL was estimated. Based on the assumption that the kinetics of luminol-amplified chemiluminescence (LCL) of stimulated PMNL possesses a time-probabilistic nature, this kinetics was described with three components. These components, obtained from different investigated systems, were analyzed and a conclusion was made that the first and the second component represent the processes resulting in extra-and intracellular myeloperoxidase (MPO)-dependent light emission (AOS generation), respectively. The second component was found to be completely dependent on the stimulus ingestion. The third component was not completely MPO-dependent and complicated for interpretation. This component was weakly dependent on the stimulus ingestion, and presents at least some intracellular processes different from those presented by the second component. A conclusion is made that the examined approach for analysis of LCL kinetics allows an assessment of extra-and intracellularly generated quantities of AOS by stimulated PMNL. The assessment could be done for emitting systems in which no additional modifications are used.

A new approach for analysis of chemiluminescent kinetics of activated phagocytes in blood.

Luminol chemiluminescence (LCL) is a simple, sensitive and time-saving tool to elucidate the oxidative activity of polymorphonuclear leukocytes (PMNL). In this study, a new approach for analysing the LCL kinetics, recorded from stimulated PMNL, and for a more accurate elucidation of their functional state, is proposed. This approach is based on the proposal that the LCL kinetics of stimulated PMNL is a result of time-probabilistic nature of the processes, leading to light emission. On this basis the LCL response was described by the parameters of a Poisson-type distribution. Relationships between these parameters and some factors influencing LCL kinetics (number of PMNL and erythrocytes, temperature, stirring) were investigated. The model descriptions of the LCL responses obtained had two components (modes). Their development in time was different. It was suggested, that the two phases of the model LCL response were connected with extracellular and intracellular LCL. The terms and the advantages of this approach for analysis of phagocyte oxidative capacity are discussed.

Stress induced tyrosine aminotransferase activity via glucocorticoid receptor.

Antiglucocorticoid RU486 was used to prove that stress induced tyrosine aminotransferase (TAT) activity is mediated via glucocorticoid receptor (GR) in the rat liver. Oral dose of RU486 (10 mg/kg) occupied liver cytosol GR in 5 min and concomitantly blocked stress (15 min immobilization) induced enzyme activity. Post-stress increase of TAT activity was observed when RU486 was administered 10 min after stress exposure. A minimal increment of 15.2 micrograms/dl plasma corticosterone ("B") lasting for 25 min is necessary for TAT induction. In the intact organism this is the time interval necessary for "B" released by external stimuli to induce synthesis of the enzyme TAT in the rat liver via GR.

Dependence of whole blood luminol chemiluminescence on PMNL and RBC count.

Luminol chemiluminescence (LCL) of whole blood as response to zymosan stimulation was used to assess the activity of polymorpho-nuclear leukocytes (PMNLs). Since PMNL and red blood cells (RBCs) are the primary components determining LCL response, the influence of PMNL and RBC count on the response was investigated over wide ranges of cell count and time interval. A linear relationship was found between maximum LCL intensity (Imax) (reflecting PMNL activity) and PMNL count. This allows a proper correction of Imax to be made as related to the sample PMNL count. This correction was valid for PMNL concentrations from 3.10(3) PMNL/ml to 6.10(5) PMNL/ml. When concentrations range from 6.10(3) PMNL/ml to 3.10(4) PMNL/ml the relationship was valid for a time interval round the peak. During the same interval the inhibitory effect of RBC count was studied. A correction of LCL was found possible for different RBC concentration ranging from 2.5.10(6) RBC/ml to 10(8) RBC/ml. These independent corrections of LCL response to PMNL and RBC counts were applied to diluted whole blood samples. The common correction of Imax was applicable in dilutions not less than 1:50. To correct light intensity for the time interval around Imax, the dilutions used had to be not less than 1:200. The results obtained permit an objective assessment of PMNL activity to be made by testing whole blood samples from different individuals and/or samples diluted to different extents.

TRH analogs with 4-nitro- and 4-N-butyloxycarbonylamino-1-methyl-2-pyrolylcarboxylic acid in position 1. Synthesis and biological properties.

Two new analogs of thyrotropin-releasing hormone (TRH), obtained by the replacement of the L-pyroglutamic acid residue with 4-nitro- or 4-N-butyloxycarbonylamino-1-methyl-2-pyrolylcarboxylic acid (analogs 1, 3), and three related derivatives, in which also the L-histidine residue was replaced with L-norvaline (analogs 2, 4) or L-norleucine (analog 5), were synthesized and tested for endocrine and central nervous system (CNS) activity. The replacement of the L-pyroglutamic acid residue with 4-nitro-1-methyl-2-pyrolylcarboxylic acid (analogs 1 and 2) resulted in the separation of the endocrine from the direct CNS activity. The effect of these analogs on the sleeping time, rectal temperature and breathing frequency, was either the same or greater than that of TRH. However, neither the correlation between the binding of analogs to TRH receptors in the brain nor their activity on the CNS parameters measured was found. Analogs 3, 4 and 5, containing 4-N-butyloxycarbonylamino-1-methyl-2-pyrolylcarboxylic acid in place of L-pyroglutamic acid, were inactive.

Hydroosmotic activities of arginine-vasopressins modified either in positions 1, 2 and 4 or at N-terminal extensions.

Vasopressin and its synthetic analogs were studied for their effect on transepithelial water flux in frog urinary bladder. As compared with AVP, 1-deamino-8-D-arginine vasopressin (dDAVP) was about 40 times less effective in stimulating osmotic water flow. The vasopressin analogs obtained by modification in positions 1 and 2 were: [1-(1-mercapto-4-tert-butylcyclohexaneacetic acid)] AVP (I); [1-(1-mercapto-4-methylcyclohexaneacetic acid)]AVP (II); [1-(1-mercapto-4-methylcyclohexaneacetic acid)-2-O-methyltyrosine]AVP (III); and those modified in position 4 were: [1-(1-mercaptocyclohexaneacetic acid)-4-arginine] AVP (IV); [1-(2-mercaptopropionic acid)-4-arginine]AVP (V). Any of the above analogs did not influence basal, but antagonized vasopressin-stimulated water flux. N-terminally extended analogs of AVP: Ala-AVP (VI); Ser-Ala-AVP (VII) and Thr-Ser-Ala-AVP (VIII) stimulated osmotic water flux to the same extent in concentration 200 times higher as that of AVP. We conclude from these studies that vasopressin analogs (I-V) competitively antagonize vasopressin-stimulated hydroosmotic activity in frog urinary bladder probably at the epithelial vasotocin V1 and/or V2 receptor site. N-terminal extension of the vasopressin molecule did not influence the capacity of AVP to induce V2 receptor-mediated action, even when used at higher concentrations.