[Synthesis of Circular DNA Templates with T4 RNA Ligase for Rolling Circle Amplification].

Currently, isothermal methods of nucleic acid amplification have been well established; in particular, rolling circle amplification is of great interest. In this approach, circular ssDNA molecules have been used as a target that can be obtained by the intramolecular template-dependent ligation of an oligonucleotide C-probe. Here, a new method of synthesizing small circular DNA molecules via the cyclization of ssDNA based on T4 RNA ligase has been proposed. Circular ssDNA is further used as the template for the rolling circle amplification. The maximum yield of the cyclization products was observed in the presence of 5-10% polyethylene glycol 4000, and the optimum DNA length for the cyclization constituted 50 nucleotides. This highly sensitive method was shown to detect less than 10^(2) circular DNA molecules. The method reliability was proved based on artificially destroyed dsDNA, which suggests its implementation for analyzing any significantly fragmented dsDNA.

[Clinical and Functional Phenotypes of Chronic Obstructive Pulmonary Disease, the Effect on the Severity of the Disease and Prognostic Value in Assessing the Risk of Disease Progression].

Background: Chronic obstructive pulmonary disease (COPD) is characterized by progressive limitation of airflow rate, hyperergic inflammatory response of the respiratory tract, and systemic manifestations. Prognosis of the disease depends on the severity of these pathogenetic components. FEV1 which characterizes the speed limit airflow do not allow predicting the rate of COPD progression. Aims: Comparison of the prognostic significance of such clinical parameters as frequency of exacerbations and the development of comorbid diseases to assess the nature of COPD progression by using different classification approaches. Materials and Methods: The prospective comparative study included 98 patients with COPD. In the framework of the study protocol, 2 visits were required when a practitioner recruited patients who met inclusion/exclusion criteria, obtained the signed informed consent, collected the anamnestic data, and performed basic procedures of the study: spirometry, 6-minute stepper test, assessment of dyspnea on questionnaire mMRC, body plethysmography, lung diffusion capacity study, dopplerechocardiography, tomography of the chest. Visit 2 was conducted in 12 months after the first one to assess the dynamics of the disease. The dynamics of the disease was considered negative if, upon repeated examination, the patient was referred to the group with more severe COPD. Results: Our study demonstrates that comprehensive assessment of such factors as the frequency of COPD exacerbations in the preceding 12 months and the presence of comorbid diseases in a patient is reasonable for assessment of disease severity and determination of disease prognosis. At the same time the frequency of COPD exacerbations as one of the evaluated factors is most strongly associated with disease progression. Conclusions: Thus, a practitioner is recommended to use the proposed additional clinical criteria to assess the severity and degree of progression of COPD.

[Catheterization of the Eustachian tube in the combined treatment of exudative otitis].

The objective of the present study was to determine the amount of a medicinal agent entering the tympanic cavity during catheterization of the Eustachian tube in 26 patients presenting with exudative otitis and admitted for its treatment to N.I. Pirogov Clinical Hospital No 1, Moscow. The patients were managed by combined therapy in conjunction with catheterization of the Eustachian tube using radiocontrast matter (the iodine containing (76%) preparation trazograf having viscosity identical with that of dexamethasone). The administration of trazograf was followed by multispiral CT of the temporal bones. It was shown that trazograf penetrated into the tympanic cavity during catheterization of the Eustachian tube and accumulated inside it in 19.2% and 77% of the cases respectively. This finding suggests that the therapeutic effect in the patients presenting with exudative otitis is due not only to the direct action of the pharmaceutical product being used on the site of inflammation but also to the restoration of the draining and aerating functions of the course of Eustachian tube catheterization.

[The clinico-psychopathological characteristics of patients with panic disorders and the efficacy of clomipramine therapy]. / Kliniko-psikhopatologicheskie kharakteristiki bol'nykh s panicheskimi rasstroistvami i éffektivnost' terapii klomipraminom.

The patients with panic affections (PA) treated with clomipramine hydrochloride (anafranil) were examined clinically and psychopathologically in relation to the drug response. PA diagnostic category was not uniform and may be subdivided into at least two subgroups, responders and non-responders to clomipramine hydrochloride. The responders had minor symptoms of PA, less severe vegetative impairment, though they were more seriously affected by depersonalization, maniophobia and agoraphobia. In this group the drug induced improvement and reduction of the attack frequency and severity within treatment month 1. Nonresponders had distinct vegetative symptomatology in the absence of agoraphobia. They failed clomipramine hydrochloride treatment as the panic attacks resumed, occasionally even with greater frequency.