[Development of early diagnosis of Parkinson's disease and comprehensive economic analysis of the effect of its implementation]. / Razrabotka rannei diagnostiki bolezni Parkinsona i kompleksnyi ekonomicheskii analiz effekta ot ee vnedreniya.

The paper summarizes the literature and author's data on the development of early (preclinical) diagnosis of Parkinson's disease (PD). Implementation of this diagnosis will promote the use of preventive therapy and change investments in diagnosis and treatment of patients. The paper declares that at present the only approach to early diagnosis of PD is positron-emission tomography of the nigrostriatal dopaminergic system, but it cannot be used for preventive examination due to its high cost. The authors consider that a less specific, but more promising approach to the development of early diagnosis of PD is the search for markers in body fluids, mainly in the blood, in patients at the prodromal stage of PD. Indeed, a number of markers as changes in the level of metabolites of monoamines, sphingolipids, urates, and indicators of oxidative stress were found in patients selected for the risk group of the prodromal stage of PD, according to characteristic premotor symptoms. In addition, it is assumed that the search for blood markers at an earlier - pre-prodromal stage is possible only in animal models of PD at the early preclinical stage. This approach can also be used to verify blood markers identified in patients at the clinical stage of PD. It is also evident that the complex socio-economic factors influencing the incidence of PD is different in developed versus developing countries. The societal and medical costs of Parkinson's are huge and efforts to improve early preclinical diagnosis of PD will lead to considerable economical and societal benefits. For instance this will allow efficient selection of patients for preclinical diagnostic tests. To assess the effectiveness of this strategy considering the uncertainty of socio-economic issues, a modification of the «cost-utility¼ analysis is proposed. For the first time, a Markov model of PD including preclinical diagnostic tests and possible neuroprotective therapy was developed and studied. Analytical outcomes of this process suggest that the idea of developing a new multimodal strategy is promising from a socio-economic point of view.

[Development of early diagnosis of Parkinson's disease based on the search for biomarkers such as premotor symptoms and changes in blood]. / Razrabotka rannei diagnostiki bolezni Parkinsona na osnove biomarkerov v vide premotornykh simptomov i izmenenii v krovi.

OBJECTIVE: To determine changes in the chemical composition of blood plasma in subjects at risk of Parkinson's disease (PD) at the prodromal stage compared with age control. MATERIAL AND METHODS: Subjects at risk were selected for the presence of characteristic premotor symptoms, including impairments of sleep, olfaction and constipation.The risk group included 12 people, the control group - 8 people. RESULTS: Among seven catecholamines and their metabolites detected in the blood, only the concentration of L-dioxiphenylalanine (L-DOPA) changed (decreased) in subjects at risk compared with the control. A decrease in the concentration of L-DOPA is considered as a manifestation (marker) of selective degeneration of central and peripheral catecholaminergic neurons in PD. In contrast to L-DOPA, the concentration of seven of the twelve detected sphingomyelins in the blood of the subjects at risk increased. Given that a change in the metabolism of sphingomyelins is associated with processes such as apoptosis, autophagy, and synucleinopathy, an increase in their concentration in the blood of patients at risk is considered as a manifestation of systemic general degeneration of central and peripheral neurons. Finally, in the blood of subjects at risk, we found a trend towards a decrease in the concentration of urates, which are endogenous neuroprotectors. CONCLUSION: The changes in the level of L-DOPA, sphingmyelins and urates in the blood of subjects at risk may serve as diagnostic markers of PD at the prodromal stage.

[Detection of effective treatment of amnestic mild cognitive impairement with cereton by testing of lipids markers]. / Issledovanie éffektivnosti tseretona pri miagkom kognitivnom snizhenii amnesticheskogo tipa na osnove testirovaniia markerov lipidnoi prirody.

AIM: Determination of effectivity and safety of Cereton (Choline alfoscerate, production by Sotex) 1200 mg/day in the treatment of cognitive functioning disorders in patients with amnestic mild cognitive impairment (aMCI) and determining its influence in the process (after a 3 month course of taking the drug) and 3 months after the end of treatment of aMCI on the change in the content of phosphatidylcholine, sphingomyelin, ceramide-metabolite sphingolipids and the activity of genes controlling the synthesis of enzymes, which control ithe metabolism of sphingomyelin and ceramide (sphingomyelinase and ceramidase). MATERIAL AND METHODS: The study involved a group of elderly patients (20 people), consisting of 14 women and 6 men, aged 51 to 82 years (mean age 70.3±9.1 years). The patients' condition met the criteria for diagnosing aMCI syndrome. Analysis of phosphatidylcholine, sphingomyelin and ceramide in the blood plasma of patients was carried out by thin layer chromatography, expression of sphingomyelinase and ceramidase genes by RtPCR. RESULTS AND CONCLUSION: A sharp increase in the content of phosphatidylcholine and ceramide, the product of sphingomyelin hydrolysis, was detected. Expression of genes (acidic sphingomyelinase and ceramidase), controlling the metabolism of ceramide, is significantly reduced in the majority of patients in the treatment with ceretone. An increase in the level of phosphatidylcholine and a decrease in the expression level of the ceramide metabolism genes during treatment with ceretone and other drugs that affect the metabolism of phosphatidylchodine and sphingolipids can be used as markers of the effectiveness of therapy.

[The potential role for sphingolipids in neuropathogenesis of Alzheimer's disease].

The review discusses the functional role of sphingolipids in the pathogenesis of Alzheimer's disease. Certain evidence exist that the imbalance of sphingolipids such as sphingomyelin, ceramide, sphingosine, sphingosine-1-phosphate and galactosylceramide in the brain of animals and humans, in the cerebrospinal fluid and blood plasma of patients with Alzheimer's disease play a crucial role in neuronal function by regulating growth, differentiation and cell death in CNS. Activation of sphingomyelinase, which leads to the accumulation of the proapoptotic agent, ceramide, can be considered as a new mechanism for AD and may be a prerequisite for the treatment of this disease by using drugs that inhibit sphingomyelinase activity. The role of sphingolipids as biomarkers for the diagnosis of the early stage of Alzheimer's disease and monitoring the effectiveness of treatment with new drugs is discussed.

[Features of memantine action profile in cholinergic deficit and intracerebral posttraumatic hematoma (hemorrhagic stroke) models in rats].

Memantine, a low-affinity non-competitive antagonist of glutamatergic NMDA-subtype receptors, was used at a daily dose of 1 mg/kg over 10 days for the treatment of rats with cholinergic deficit induced by the chronic administration of scopolamine (1 mg/kg, 20 days). The drug prevented violation of the learning of conditioned active and passive avoidance reflexes and produced no significant effect on the emotional state of animals in elevated plus maze (EPM) test. In animals with intracerebral posttraumatic hematoma (hemorrhagic stroke), memantine (2 mg/kg, for 3 days after operation) completely prevented the loss of animals, reduced the neurological deficit, improved conditioned passive avoidance reflex performance, and decreased emotional stress in the EPM test.

[Relationship of tumor necrosis factor alpha expression with activation of sphingomyelinase and lipid peroxidation after removal of cholestatic factor].

The goal of this work was to study the expression of tumor necrosis factor alpha (TNFalpha), sphingomyelin cycle activation, and lipid peroxidation (LPO) processes after the removal of a cholestatic factor in the liver subjected to different durations of cholestasis. Restored bile flow after a 9-day hepatic cholestasis normalized sphingomyelinase (SMase) activity and levels of TNFalpha and LPO products. The removal of a cholestatic factor after a 12-day cholestasis did not normalize the studied parameters: SMase activity and the levels of TNFalpha and LPO products remained much higher compared to control. A significant positive correlation between TNFalpha expression, SMase activity, and LPO rate has been revealed. The obtained data indicate that hepatocyte apoptosis after bile outflow restoration in late cholestasis can be due to the activation of the sphingomyelin cycle, LPO, and TNFalpha expression. The synergistic interaction can sharply increase the proapoptotic capacity of each of these factors since TNFalpha activates SMase and LPO, SMase activity depends on the LPO rate, while ceramide, an SMase-produced secondary messenger of apoptosis, can induce oxidative stress.

[The influence of tumor necrosis factor alpha on the processes of sphingomyelin cycle and lipid peroxidation in brain].

The influence of tumor necrosis factor alpha (TNF-alpha) on the processes of sphingomyelin cycle activation and intensity of peroxidation in animal brain in vivo has been studied. Alterations in activity of sphingomyelinase, a key sphingomyelin cycle enzyme and in sphingomyelin, ceramide content as well as accumulation of the products of lipid peroxidation (diene conjugates and diene ketons) were measured in the cortex, the cerebellum and the hippocampus of rats 5, 15, 30 min, 1, 2 and 5 hours after TNF-alpha intraperitoneal injection in dosage 100 mkg per animal. It is shown that 2 hours after the injection, TNF-alpha initiated an accumulation of the products of lipid peroxidation, which intensively developed in the cerebellum and the hippocampus. Sphingomyelinase activation was found in the same brain structures. At the initial stage of TNF-alpha action, an increase of lipid peroxidation products correlated with sphingomyelinase activation in the cerebellum and the hippocampus suggesting an interaction of two cell signal systems of sphingomyelin cycle and oxidative system.

[Changes in sphingomyelinase activity, tumor necrosis factor alpha level, and lipid peroxidation rate in the course of development of cholestatic liver injury].

Changes in sphingomyelinase activity, tumor necrosis factor alpha expression, and lipid peroxidation rate in the course of development of cholestatic liver injury have been studied. The same type phase shifts in the parameters analyzed were observed, which included a marked decrease at the early stages of cholestasis (days 3-6) and a pronounced increase at the later stages (days 12-16), i.e., under the conditions of developed pathology. There is a significant positive linear correlation between tumor necrosis factor alpha expression, sphingomyelinase activity, and lipid peroxidation rate during cholestatic injury. The changes detected may reflect balance between the effects of the two major bile components--bilirubin, which is accumulated in the liver at the early stages of cholestasis, and bile acids, whose influence dominates at the later stages of pathologic process. Our results indicate that tumor necrosis factor alpha overexpression, the sphingomyelin cycle activation, and lipid peroxidation intensification may cause apoptosis of hepatocytes at the late stages of cholestasis.

Content and structure of ceramide and sphingomyelin and sphingomyelinase activity in mouse hepatoma-22.

The relative content of phosphatidylcholine is lower and that of sphingomyelin is higher in transplantable fast growing mouse hepatoma-22, thus decreasing their ratio approximately 2.5-fold versus normal liver. The ceramide content and the neutral sphingomyelinase activity is markedly higher (3- and 6.5-fold, respectively), whereas the acid sphingomyelinase activity is 4-fold lower in hepatoma-22 versus normal liver. The content of saturated fatty acids in ceramide and sphingomyelin of hepatoma-22 is higher than in normal liver. All sphingolipids of hepatoma-22 contain a considerable amount (25-37%) of sphinganine (dihydrosphingosine) along with sphingenine (sphingosine), whereas sphingolipids of normal liver contain predominantly sphingenine (over 95%). These results indicate that the activity of enzymes involved in sphingolipid biosynthesis and catabolism is disturbed in the transplantable mouse hepatoma-22 compared to normal liver.

[Changes in the activity of neutral and acidic isoforms of sphingomyelinase in hepatoma-22, regenerating and ischemic liver]. / Izmenenie aktivnosti neitral'noi i kisloi izoform sfingomielinazy v gepatome-22, v regeneriruiushchei i ishemizirovannoi pecheni.

Activity of neutral and acidic sphingomyelinases (N- and A-SMases) were studied in regenerating liver after partial hepatectomy (during 48 hrs after operation), in ischemic liver during 15, 30 min and 1 and 2 hrs ischemia and during following reperfusion (from 5 min up to 2 hrs), in hepatoma- 22 after 15 days of transplantation and in liver of tumor bearing animals. It was shown that activity of N-SMase is increased in hepatoma-22 and in regenerating liver and it is decreased in ischemic liver. Following reperfusion of ischemic liver area activity of enzyme was found to have returned to baseline in dependence on time of ischemia and reperfusion. Activity of A-SMase is decreased in tumor, is not changed in regenerating liver and increased after long time of ischemia. It was supposed that N-SMase is involved in cell proliferation, but A-SMase is connected with cell damage.

TNF alpha-induced aerobic synthesis of ATP on plasma membranes of target cells. The relation to the expression of the nuclear oncogene c-myc.

The accumulation of ATP by preparations of plasma membranes enriched particles (PMEP) isolated from rat hepatocytes, murine splenocytes and human T-lymphocytes has been investigated after the binding of human and murine tumour necrosis factors (TNF alpha) to their specific receptors. The TNF alpha-induced expression of the nuclear oncogene c-myc in intact hepatocytes has been also studied. TNF alpha induced the marked biosynthesis of ATP on PMEP of hepatocytes and splenocytes within the first minute of incubation. The biosynthesis of ATP was independent of the activity of adenylate kinase and only occurred in the presence of all the components of aerobic phosphorylation and the electron acceptor, cytochrome C or diferric transferrin. The level of ATP on PM correlated with the degree of expression of the nuclear oncogene c-myc in the same target cells. Adriamycin totally suppressed the biosynthesis of ATP on PM and simultaneously inhibited the expression of c-myc. The ATP synthesized on PM is suggested to be involved in transduction of the proliferative or growth signal to the cell nucleus.

[The role of sphingomyelin cycle products in development of apoptosis mediated by Fas receptors and tumor necrosis factor alpha]. / Rol' produktov sfingomielinovogo tsikla v razvitii apoptoza, indutsirovannogo cherez retseptory Fas, i faktora nekrosa opukholi al'fa.

Programmed cell death or apoptosis is a cell suicide developing according to a specific program, in which the sphingomyelin cycle products, ceramide and sphingosine, play the central role. The present review provides published data and the authors' results suggesting that the content of ceramide and sphingosine in the cell is controlled by the tumor necrosis factor alpha and activators of Fas receptor. The results of many experiments confirmed that the sphingomyelin cycle products induce cells death by the apoptotic pathway and enhance induced apoptosis. Ceramide and sphingosine regulate the activity of enzymes involved in transduction of apoptosis signal (protein kinases. Phosphatases, and proteases) and act as second messengers in transduction of the apoptosis signal.

[Correlation of cytotoxic activity of mutant forms of tumor necrosis factor alpha with changes in the level of free sphingosine in murine liver]. / Korreliatsiia tsitotoksicheskoi aktivnosti mutantnykh form faktora nekroza opukholei-alpha s izmeneniem urovnia svobodnogo sfingozina v pecheni myshei.

Sphingosine, the product of enzymatic degradation of sphingomyelin, displays a high cytotoxic activity and is accumulated in animal organs under the action of the tumour necrosis factor (TNF alpha). To elucidate the role of sphingosine in the realization of TNF cytotoxicity, TNF mutants were obtained which differed in their cytotoxic action on L929 cells. The wild strain of TNF and the mutant having a deletion in position 67-71 displayed the highest toxicity and sharply stimulated sphingosine accumulation in mouse hepatocytes. A moderate increase in the sphingosine content was induced by mutants with point and double mutations in positions E127Q, I155L and V150I displaying a much lower toxicity in comparison with the wild strain. The toxic, mutagenic and antimutagenic activities of sphingosine were investigated. Despite the high degree of cytotoxicity, sphingosine did not display any mutagenic activity but had a pronounced antimutagenic effect on E. coli cells. The role of phospholipid enzymatic degradation products in activation of sphingomyelin cycle enzymes stimulating of sphingosine accumulation in animal cells under the action of TNF alpha is discussed.

[Modulation of the level of sphingomyelin cycle products and expression of the CD3 receptor in immunocompetent organs by fumonisin B1]. / Moduliatsiia fumonizinom B1 soderzhaniia produktov sfingomielinovogo tsikla i ékspressii retseptora CD3 v immunokompetentnykh organakh.

The fungi (Fusarium moniliforme) residing on cereals produce a broad range of mycotoxins, among which fumonisins display a high toxicity alongside with carcinogenic and teratogenic activities. Taking into account the ability of fumonisins to inhibit sphingolipid synthesis, the role of sphingomyelin cycle products in immune reactions was studied with the view of establishing the correlation between the expression of the surface receptor CD3 in immunocompetent organs (spleen, thymus) (T-cell mediated immunity) and the degree of sphingomyelin cycle activation (changes in the activities of sphingomyelinase and sphingomyelin and ceramide content) in the spleen, thymus and liver 2.5 hours after intraperitoneal injection of fumonisin B1 (FB1) (5 and 20 micrograms/animal). Significant sphingomyelinase activation was found in the thymus of animals injected with 20 micrograms of fumonisin. It coincides with a loss of the sphingomyelin and ceramide content. The changes in the sphingomyelinase activity and sphingomyelin content in the spleen and in the liver caused by fumonisin were insignificant, while the ceramide content dropped drastically. Fumonisin decreased the receptor CD3 expression on the surface of thymus cells "in vitro" and "in vivo", which is consistent with the sharp decrease of the ceramide content in this organ. Ceramide accumulation in thymus and spleen cells treated with sphingomyelinase in vitro correlates with the increased affinity of receptor CD3. The putative role of ceramide in the expression of receptors modulating T-cell mediated immunity under the influence of fumonisin is discussed.

[Correlation of DNA degradation, induced by tumor necrosis factor-alpha, with accumulation of sphingosine in the nucleus and peroxides in DNA]. / Korreliatsiia degradatsii DNK, indutsirovannoi faktorom nekroza opukholi-alpha, s nakopleniem sfingozina v iadre i perekisei v DNK.

The tumour necrosis factor alpha (TNF-alpha) initiates DNA degradation in many types of cells, eventually resulting in a programmed cell death (apoptosis). In order to study the mechanism of TNF-alpha action in vivo, the dynamics of mouse liver DNA fragmentation was examined after intraperitoneal administration of recombinant hTNF-alpha (10 and 40 micrograms per animal). The number of single-strand (SSB) and double-strand (DSB) breaks was determined electrophoretically and interruption of hydrogen bonds (secondary structure defects SSD) were studied by the kinetic formaldehyde method. In parallel experiments the accumulation of peroxide products in DNA, the activity of sphingomyelinase and the content of sphingosine in liver cell nuclei were measured. Sphingomyelinase activation and sphingosine accumulation in the nuclei were found to coincide in time with the maximal values of DNA degradation parameters (SSB, DBS and SSD). TNF-alpha caused a dose-dependent accumulation in DNA peroxide products which seem to lead to the DNA damages. The role of sphingomyelin cycle products and peroxidation in DNA fragmentation induced by TNF-alpha in vivo is discussed.

[Change in the level of sphingosine in rat liver nuclei and cells during oncogene superexpression induced by cycloheximide]. / Izmenenie urovnia sfingozina v iadrakh i kletkakh pecheni krys pri indutsirovannoi superékspressii onkogenov tsiklogeksimidom.

Changes in the sphingosine content in rat liver cells and nuclei have been studied with reference to the level of nuclear oncogene expression, induced by cycloheximide (0.1, 0.5 and 3.0 mg/kg). It has been found that only the sublethal (3 mg/kg) dose of cycloheximide which induces the superexpression of c-fos and c-myc oncogenes can promote sphingosine accumulation in the cell. At the moment of enhanced expression of nuclear oncogenes, the maximum content of free sphingosine exceeds the control level 1.5- and 3-fold in the cell and in the nuclei, respectively. The difference in the sphingosine accumulation patterns in the cell and in the nuclei testifies to the fact that sphingomyelin metabolism is more active in the nuclei than in the cell. Sphingosine accumulation in the nuclei is characterized by coordination of sphingomyelinase activity and changes in the sphingomyelin content. A comparative analysis of activities of enzymes of sphingomyelin (sphingomyelinase) and phosphatidyl inositol (phosphatidyl inositol kinase) cycles indicates that in the nuclei the activation of the sphingomyelin cycle forestalls the cycloheximide-induced activation of the phosphatidyl inositol cycle and the maximal accumulation of nuclear oncogene mRNAs. A model of activation of oncogene expression with participation of sphingosine inhibiting protein kinase C and activating casein kinase II, the key enzymes of the signal transduction system of cell proliferation and differentiation, is proposed.

[The effect of tumor necrosis factor on the free sphingosine level and sphingomyelinase in murine liver cells and nuclei]. / Vliianie faktora nekroza opukholi na uroven'svobodnogo sfingozina i aktivnost' sfingomielinazy v kletkakh i iadrakh pecheni myshei.

The effects of the human recombinant tumour necrosis factor (TNF) (10 and 40 mg/kg of body mass) on sphingomyelinase activity and sphingosine content in mouse (C57bl) liver cells and nuclei have been studied. Whereas sphingomyelinase is known to be a key enzyme of sphingomyelin metabolism, sphingosine, being a product of deep enzymatic hydrolysis of sphingomyelin, controls the activity of various phosphokinases. The primary response of liver cell to TNF consists in the inhibition of sphingomyelinase; its activation occurs at later periods: after 2 hours at 10 mg/kg TNF and after 4 hours at 40 mg/kg TNF. In the nucleus activation of sphingomyelinase is observed within the first 60 min after TNF administration. Sphingosine accumulation in mouse liver cells and nuclei coincides in time with sphingomyelinase stimulation. In the nuclei activation of the sphingomyelin cycle by TNF is far more pronounced than in the cells, being observed at early periods after TNF injection. A signal mechanism of TNF action on mouse liver cells and nuclei involving a TNF-specific receptor and sphingosine which may activate this receptor phosphorylation is discussed.

[Change in the level of endogenous sphingosine in cell nuclei from regenerating rat liver]. / Izmenenie urovnia éndogennogo sfingozina v iadrakh kletok regeneriruiushchei pecheni krys.

High-performance liquid chromatography was used to study the changes in the sphingosine content in regenerating rat liver cell nuclei during RNA and DNA synthesis. It was found that activation of nucleic acid synthesis was accompanied by sphingosine accumulation in cell nuclei in parallel with the induction of the sphingomyelin cycle consisting in the increasing activity of sphingomyelinase and alteration of the sphingomyelin and ceramide content. To clarify the mechanism of sphingosine involvement in replication and transcription, the ability of this product to interact with DNA and modify the activity of RNA-polymerase in vitro was studied. At 10(-4) M sphingosine prevented the interaction of acridine orange with DNA and activated the transcription enzymes. Several alternative mechanisms of sphingosine involvement in the control of nucleic acid synthesis are discussed.

[Comparison of lipid metabolism in rat liver cells and nuclei during cycloheximide-induced superinduction of nuclear oncogenes]. / Sravnenie metabolizma lipidov v iadrakh i kletkakh pecheni krys v usloviiakh TsGI-indutsirovannoi superékspressii iadernykh onkogenov.

Using a model of cycloheximide (CHI)-induced expression of nuclear oncogens, a comparative study of metabolism of the major lipid classes in rat liver nuclei and cells was carried out. A short-term activation of sphingomyelinase which preceded on a time scale the maximal accumulation of c-fos and c-myc transcripts was observed both in the cells and in the nuclei. In contrast with the whole cell, the level of phospholipase C activity in the nuclei did not change under conditions of oncogene activation. It was found that the maximal expression of nuclear oncogens coincided in time with cyclic changes in the content of practically all phospholipids and neutral lipids with simultaneous activation of their synthesis both in the cells and in the nuclei. However, in the nuclei the sphingomyelin metabolism activation was predominant. It is concluded that in the nucleus sphingomyelin and its metabolites may influence oncogene expression via nuclear protein kinase C.