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St. Cercet. Virusol. 34 (1) 2004


Part I

Part II


33 - The implementation of the molecular diagnosis in Romania in order to estimate the prevalence of the sexually transmitted infections induced by Mycoplasmas and Chlamidia trachomatis
Constanta Antipa
Stefan S. Nicolau Institute of Virology, Bucharest, Romania
Abstract. The objectives of the project lie in the necesity of an epidemiological assesment of the incidence of sexually transmitted infections induced by mycoplasmas and Chlamydia trachomatis, germs often ignored, although responsible of severe acute and chronic infections, such as pelvic inflammatory disease (PID), infertility, perinatal diseases, etc.
The project tries to optimize the diagnosis of these diseases and to introduce into it molecular techniques, aimed especially at the asymptomatic forms of disease.
The objectives of the project will be achieved by: formation of groups of patients according to the age, the sex and the symptomatology; establishment of criteria regarding the sampling, processing and stockage of the specimens taken from the patients; evaluation of the incidence of infections with Chlamydia trachomatis and Mycoplasma genitalium, in dependence on the results obtained by tests performed by usual techniques(EIA, IFD, isolation and identification of mycoplasmas). In the case of positive results, the susceeptibility to antibiotics will be tested; estimation of immunity of patients to the mentioned germs by performing serological tests; optimization and standardization of PCR protocols, in order to make possible their introduction into the current diagnosis of sexually transmited infections caused by Chlamydia trachomatis and Mycoplsma genitalium; interpretation of results, their correlation with the clinical data, final assessment of the prevalence of STD induced by Chlamydia trachomatis and Mycoplasma genitalium in symptomatic and asymptomatic patients.
In view achieving the project, we intend to select on the basis of clinical and laboratory criteria a number of patients with STD, beside patients without symptoms specific of these diseases. The biological specimens taken from these patients will be: endocervical swabs, urine, blood; they will be processed and stocked in compliance with the established protocols in order to be tested according to the algoritm elaborated by us.

Keywords: sexually transmited diseases, chlamydia trachomatis, mycoplasmas genito-urinary, PCR.

37 - DNA methylation pattern in cervix cancer - molecular biomarker for monitoring and prognostic
Gabriela Anton
Stefan S. Nicolau Institute of Virology, Bucharest, Romania
Abstract. DNA methylation is essential for mammalian cell development, representing one of the numerous signals involved in the cell cycle regulation. One of the most important properties of CpG islands from mammalian cell promoters is represented by their unmethylated pattern in the germ line and indeed in most somatic cells. CpG islands act often as strong promoters and some authors consider them as replicating origins. Gene expression is regulated not only by DNA sequence but also by its epigenetic status. Recent studies exhibited that aberrant promoter methylation of some tumorsupressor genes affect their silencing and correlate with carcinogenic initiation and progression. This paper is focused on studies concerning some epigenetic changes in biologic samples from patients with cervix cancer. As we know, in Romania, these molecular aspects of cancer diseases in general (and in special of cervix cancer) have not been investigated yet. The objectives of this paper are: i) the analysis of some tumorsupressor genes (p16, PTEN, HIC-1, DAPK) expression levels in biological samples from control and patients with cervix cancer; ii) the selection of the adequate protocols, in order to evaluate the tumorsupressor gene promoters hyper-methylation; iii) the correlation between methylation pattern of multiple genes and cervical carcinogenesis status.

Keywords: epigenetics, DNA methylation, cervix cancer.

43 - The assessment of different culture conditions on replicative and functional capacity of hematopoietic stem cells
Mihaela Chivu
Stefan S. Nicolau Institute of Virology, Bucharest, Romania
Abstract. Hematopoietic stem cells represent a renewable source of material for cellular therapy. Therefore, the development of ex vivo experimental systems for studies on hematopoietic stem cells is very important. The promise of stem cells for clinical therapy, however, is limited because the number of cells that can be obtained is inadequate. Manipulation of the in vitro culture microenvironment, primarily through cytokine supplementation, has been the predominant approach in studies attempting to expand primary human hematopoietic stem cells in vitro. While promising results have been accomplished, it seems obvious that novel methods must be developed before cellular therapies using these stem cells can become standard operation. This study intend: i) to design a device for efficient cord blood collection and to evaluate the obstetric factors that may influence the final collection yield in terms of recovered cell number; ii) to develop novel methods able to sustain the increase of hematopoietic stem cell number; iii) to assess the susceptibility of hematopoietic stem cells to different viral infections; iv) to create a system for toxicity studies following exposure to anti-neoplasic and anti-viral agents.

Keywords: hematopoietic stem cells, culture systems, application

49 - The study of the pleiotropic effects of treatment with statins.
M. Cornitescu1, Teodora Cornitescu1, C.E. Cernescu1, Angela Mirela Gaveanu2, C. Popa2
1Ştefan S. Nicolau Institute of Virology, Bucharest, Romania2 Vlad Voiculescu Institute of Cerebrovascular Diseases, Bucharest, Romania
Abstract. Large studies have demonstrated that statin therapy reduces the risk of cardiovascular morbidity and mortality. For a long period of time it has been considered that the principal responsible mechanism for this effect is the inhibition of cholesterol metabolism. The remodelling of the atheroma plaque takes place after more than 2 years of therapy, while improvement of patients’ status appears much earlier, which suggests the existence of secondary mechanisms of action of statins, complementary to that of action on cholesterol metabolism. Experimental data suggest that statins have a specific effect of inhibition of inflamation and, as a consequence, therapeutic effects in cardiovascular diseases, type 2 diabetes and neurodegenerative diseases. There are also experimental data showing that these drugs have antiviral effects by altering cholesterol metabolism. Current data support the necessity of continuing the investigation on the pleiotropic effects of statins.

Keywords: statins, inflammation, type 2 diabetes, neurodegenerative diseases, viral infection.

57 - Features of the immune and hormonal response in children viral neuroinfections.
Florica Toparceanu, Irina Alexiu, Carmen C. Diaconu, Coralia Bleotu
Stefan S. Nicolau Institute of Virology, Bucharest, Romania
Abstract. Viral neuroinfections has one of the main places in children clinical issues. Between central nervous system (CNS) and the immune system (IS) there is a bidirectional communication, sustained mainly by hypothalamo-pituitary-adrenal axis and IS mediators. Correlation between cytokines seric amounts, cortisol levels, and clinical / ethiological data concerning patients will permit insights in non-specific immune response, therapeutically modulated, in different ethiological conditions. Viral strain identification of pathogens causing children neuroinfections is necessary for  excluding polio infection and bioterorist attack suspicions, and for estimating wild viral circulation’s impact on children infectious pathology. Tests’ results, clinical and treatment features will be gathered in a database, for improving diagnostic and therapeutic protocols.
Keywords: viral neuroinfections, immune response modulation, glucocorticoids
63 - Supplementary pathways for the explanation of neuropsychiatric disorders pathogeny.
C.N. Zaharia1, D. Popescu2, M. Stoian1
1Stefan S. Nicolau Institute of Virology, Bucharest, Romania2Centre of Animal Physiology and Neurobiology, Faculty of Biology, Bucharest University, Romania
Abstract. Depression is considered as a disease with many possible causes, although in the past it was considered as an episodic one. Usually anxiety is associated (as morbidity) with depression. Most of the patients exclusively treated for anxiety are developing a form of depression. In the medical treatment of depression and anxiety, beside the current medicines, a lot of natural products are used. Today, Hypericum Perforatum L is commercialized in different standard forms like capsules, nasal drops or hydroalcoholic extract in Germany, Great Britain and United States. In present, new approaches are developed for treating neuronal disorders. Molecular studies correlated with experiments on animals and clinical studies are currently used. New directions of study are passing from phenomenology to the molecular mechanisms in the central nervous system. Some applications of molecular dynamics and docking were developed at neuronal level. Subsequently, it is possible to investigate the mechanisms implicated into the transbilayer transport of active substances, such as hypericin, hyperforin and quercetin. Molecular dynamics permits us to obtain informations concerning (i) stability and functionality of the complex membrane-neurotransmitter, indicating the insertion of the neurotransmitter into the lipid bilayer; (ii) anchoring of small molecules at the cellular membrane, simulated by the interaction with the lipid matrix of the cellular membrane; (iii) stochastic pores apparition through the cellular membrane, as a supplementary pathway for neurotransmitters transport. These supplementary pathways are signaled at patients in stress conditions or presenting a mood disorder, such as depression. Our study concerns the description of neurotransmitter receptors into the postsynaptic membrane and of the phenomenon of drug binding during the treatment of mood disorders (depression and anxiety). This method enables identification of molecular docking surfaces: receptor-neurotransmitter and receptor-drug.

Keywords: molecular dynamics, docking, lipid bilayers, stochastic pores, depression, anxiety.