[3-Dimensional microscopy as a method for volume measurement in cells undergoing apoptosis].

Different patterns of cell volume perturbations are commonly used for modes of cell death: necrosis (cell swelling) and apoptosis (cell shrinkage). In this study we employed recently developed three dimensional microscopy for the measurement of the volume of attached vascular smooth muscle cells transfected with E1A-adenoviral protein. These cells undergo rapid apoptosis in the absence of growth factors or in the presence of staurosporine. In 30-60 min of serum deprivation the volume of these cells is increased by -40% that corresponds to the time point of maximal activation of caspase 3 and chromatin cleavage. In 10-15 min swollen cells exhibit morphological collapse indicated by formation of apoptotic bodies. In contrast to serum-deprived cells, staurosporine leads to attenuation of cell volume by 30%. In this case, apoptotic bodies are detected in -2.5 hr after maximal shrinkage. Thus, our results show that cell shrinkage can not be considered as universal hallmark of apoptosis. The role of stimulus-specific cell volume perturbation in the triggering of the cell death machinery should be examined further.

Thermal inactivation of volume-sensitive K⁺,Cl⁻ cotransport and plasma membrane relief changes in human erythrocytes.

Previously, we reported that in mammalian erythrocytes irreversible annealing of spectrin heterodimers at 49-50 °C abolished cell volume-dependent regulation of ion carriers, thus suggesting an implication of a two-dimensional (2D) membrane carcass in volume sensing and/or signal transduction. To further examine this hypothesis, we employed atomic force microscopy. This method revealed folded membrane relief of fixed human erythrocytes with an average wave height of 3-5 nm covered by globular structures with a diameter of 40-50 nm and an average height of 1-2 nm. Erythrocyte swelling caused by reduction of medium osmolality decreased the height of membrane surface waves by 40 % and increased K(+),Cl(-) cotransport by approximately sixfold. Both volume-sensitive changes of membrane relief and activity of K(+),Cl(-) cotransporter were abolished by a 10-min preincubation at 50 °C. Our results strongly suggest that volume-dependent alterations of the human erythrocyte membrane relief are caused by reorganization of the 2D spectrin-actin network contributing to regulation of the activity of volume-sensitive ion transporters.

[Antiviral activity of human recombinant gamma-interferon and recombinant hybrid protein of tumor necrosis factor-alpha-thymosin-alpha1 on models of herpes-virus and cytomegalovirus infection in vitro]. / Protivovirusnaia aktivnost' rekombinantnogo interferona-gamma cheloveka i rekombinantnogo gibridnogo belka alpha-faktor nekroza opukholei-timozin-alpha1 na modeliakh gerpes-virusnoi i tsitomegalovirusnoi infektsii in vitro.

The conducted studies showed a certain efficiency of gene-engineering preparation of IFN-gamma and TNF-T in virus infections caused by herpes simplex virus 2 (HSV-2) and cytomegalovirus (CMV) in cell cultures of human embryo fibroblast (HEF). The drugs have no viricidal action. IFN-gamma, when used according to the treatment scheme in vitro, proved to be more effective versus TMF-T both in HSV-2 and in CMV. It inhibited significantly the HSV-2 reproduction within the dilution range of 1:50 to 1:500. It was also effective, when used in CMV, within the dilution range of 1:5000 and lower, whereas TNF-T was effective within the range of 1:500 and lower as well as in 0.1 multiple infection. A significantly higher effect was ensured when the drugs were used for prevention. In HSV-2, IFN-gamma inhibited the virus reproduction, like in the treatment scheme, within the dilution range of 1:50 to 1:500, whereas TNF-T was effective in the range of 1:50. In CMV, the drugs' effect, when used for prevention, was similar to that observed in the treatment scheme. The highest inhibition values were registered for HSV-2, when it was used 24 hours before infection (IFN-gamma--2.25 Ig, dilution range of 1:50; TNF-2--1.0 Ig, dilution range of 1:50). IFN-gamma and TNF-2 exert a synergic action on different stages of virus reproduction. A reliable additive effect was ensured in prevention made 4 hour before infection by IFN-gamma and TNF-T only in experimental CMV infection.

[Correction of rate and structure of ventricular rhythm in permanent atrial fibrillation: a novel pathogenetic approach]. / Korrektsiia chastoty i struktury zheludochkovogo ritma pri postoiannoi forme mertsatel'noi aritmii: novyi patogeneticheskii podkhod.

AIM: To analyze action of digoxin and some non-digitalis drugs (beta-blockers, verapamil, amiodarone, d,l-sotalol) and their combinations on electrical activity of atria, frequency and structure of ventricular rhythm in patients with permanent atrial fibrillation. MATERIAL AND METHODS: One hundred patients with permanent atrial fibrillation and heart rate at rest above 80 bpm were divided into 9 treatment groups. High-resolution orthogonal Frank lead ECG was recorded before and after allocated treatment. Analysis included construction of ff-waves periodograms, histograms of RR interval, cardiointervalograms and application of autocorrelation function. RESULTS: It was demonstrated that frequency and form (structure) of ventricular rhythm was determined not only by the state of AV conduction but also by value of basic period of ff-waves. The mechanisms of ventricular rate deceleration by investigated drugs were not identical. beta-blockers and verapamil directly slowed AV conduction without changing parameters of ff-waves and differed from each other only in action on parameters of concealed conduction in AV node. Action of digoxin in patients with ff-waves period equal to or exceeding 0,15 s was biphasic. During phase I shortening of ff-wave period (by 0.025+/-0.012 s) occurred. This was associated with increase of their concealed conduction through AV node. The latter phenomenon represented independent mechanism of ventricular rhythm deceleration. During phase II of digoxin action direct inhibition of AV conduction took place. Amiodarone and d,l-sotalol increased basic ff-waves period. This facilitated their conduction through AV node. Greater heart rate slowing effect of d,l-sotalol was attributed to its ability to augment concealed conduction. Due to their antiarrhythmic qualities amiodarone and d,l-sotalol slowed heart rate in patients with peak RR duration in the region of 0.28-0.46 s. These patients often had bi- and tri-modal structure of interval RR histogram. Changes of ventricular rhythm structure during use of various drugs were different. Action of digoxin was most whilst that of beta-blockers least favorable. CONCLUSION: Choice of a drug for treatment of permanent atrial fibrillation should be conducted with consideration of ff-waves periodicity, parameters of RR interval histogram, and characteristics of ventricular rhythm structure.

[Medication reducing therapy of atrial fibrillation: a new approach to an old problem]. / Medikamentoznaia urezhaiushchaia terapiia pri mertsatel'noi aritmii: novyi podkhod k staroi probleme.

AIM: To study influence of different rate-reducing medicine groups on atrial electrical activity and ventricular thyrhms structure in chronic atrial fibrillation (CAF) and thus to optimize treatment. MATERIAL AND METHODS: 57 patients with CAF treated with digitalis, beta-blockers, their combinations, verapamil, amiodaron and sotalol were studied. Repeatedly highly-resolution ECG analysis was made. RESULTS: Atrial fibrillation is characterized by ff-waves basic period from 0.12 s to 0.20 s. Ventricular characteristics were determined both by AV conduction and basic ff-waves periods. The reducing of basic ff-period is the leading mechanism of digitalis rate-reducing actions in typical cases. The direct AV block appears on the late stage of therapy and is moderate. CONCLUSION: Combined therapy by medicines with different mechanisms of rate-reducing actions is the most efficient. The choice of the drug and its doses is determined by basic ff-waves periods value and the ventricular rhythm structure.

[24-hour ECG monitoring in cardiac fibrillation]. / Primenenie sutochnogo monitorirovaniia EKG pri mertsatel'noi aritmii.

AIM: To elucidate clinical value of 24-h ECG monitoring followed by analysis of cardiac rhythm structure in patients with perpetual cardiac fibrillation (CF). MATERIALS AND METHODS: 24-h ECG monitoring has been performed 163 times in 142 patients aged 33-82 years. Interval histograms and cardiointervalograms of R-R intervals were performed. RESULTS: 24-h ECG monitoring has shown the phenomena which can be used for perfection of the diagnosis and treatment: features of 24-h changes in heart rate; features of ventricular ectopy; periods of asystole; preclinical signs of hyperdigitalization; frequency-related shift of ST segment. CONCLUSION: 24-h ECG monitoring in patients with perpetual CF provides valuable diagnostic information. Therefore, it should be conducted in all of them irrespective of subjective state and routine ECG data, especially in deciding on antiarrhythmic therapy.

[The combined use of the cardiointervalogram and 24-hour ECG monitoring in analyzing heart rhythm disorders]. / Sochetannoe primenenie kardiointervalogrammy i sutochnogo monitorirovaniia EKG pri analize narushenii serdechnogo ritma.

2500 records of cardiointervalograms and 24-h ECG monitoring for 1530 arrhythmic patients have been analysed. This complex approach to analysis of cardiac arrhythmia allowed a more accurate differential diagnosis, more detailed visualization of rhythmic abnormalities, more advanced analysis of neurovegetative background of developing rhythmic shifts.

[Analysis of the structure of cardiac rhythm in atrial flutter]. / Analiz struktury serdechnogo ritma pri trepetanii predserdii.

Heart rhythm was studied by computer in 67 patients with atrial flutter of different etiology. It was discovered that respiratory activity has an influence on the conduction of atrial flutter waves to the ventricles. In the majority of cases, the conduction of atrial waves to the ventricles is multiple with a constant conduction ratio. However, in some cases the conduction of atrial flutter waves via the atrioventricular system occurs according to the Samoilov-Wenckebach periodicals. In the latter case and in atrial flutter with a conduction ratio of 4:1, one-third of patients manifest, after the recovery of the sinus rhythm, atrioventricular blockade, grade 1.

[Sinoatrial block with Wenckebach's period: diagnosis by means of automatic analysis of the cardial rhythm structure and clinical evaluation]. / Sinoatrial'naia blokada s periodami Samoilova-Wenkebakha: diagnostika pri pomoshchi avtomaticheskogo analiza struktury serdechnogo ritma i klinicheskaia otsenka.

Computer served to study by automatic analysis the structure of the cardiac rhythm in 190 patients: 123 patients after removal of the persistent form of atrial fibrillation, 24 with neuro-circulatory dystonia and 43 healthy individuals. Sino-atrial block with Samoilov-Wenkebach periods was recorded in 26 individuals. The characteristic changes of the cardiointervalogram in the given disorder of intracardiac conduction are described, which facilitates diagnosis considerably. The sino-atrial block with Samoilov-Wenkebach periods in the observed was not accompanied by clinical symptoms and exerted no untoward effects on the distant results of electro-impulse therapy in patients with persistent forms of atrial fibrillation.

[Atrial fibrillation process studied in clinical practice by using statistical analysis methods]. / Izuchenie protsessa fibrilliatsii predserdii v klinicheskoi praktike s primeneniem statisticheskikh metodov analiza.

The results of studying the electrical activity of the atria in 53 patients with cardiac fibrillation by means of a special digital computer system are discussed. On the strength of the aggregate of amplitude, rate, and morphological characteristics, three varieties of fibrillation waves are distinguished. It is shown that in the absence of cardiotropic therapy, the principal regular features in the distribution of the ff waves according to the frequency ranges are preserved in one and the same patients for a long period of time. A characteristic dynamics of changes in atrial activity during treatment with preparations of digitalis was disclosed, which may be an additional criterion of digitalization. A connection was revealed between the changes in atrial activity during treatment with digitalis preparations and the characteristic dynamics of the ventricular rhythm structure. The absence of such correlative connection may be evidence of the possible occurrence of atrioventricular block after restoration of the sinus rhythm.

[Study of the electric activity of the heart atria in auricular fibrillation with the aid of the instrumental correlation analysis]. / Issledovanie elektricheskoi aktivnosti predserdii pri mertsatel'noi aritmii s pomoshch'iu pribornogo korreliatsionnogo analiza.

The autocorrelation funtion of the electrocardiographic signal in the V1 lead is approximated by the formula of an attenuated consinusoid. Parameters are singled out (median period, attenuation factor) to characterize the atrial activity. A procedure for recording the auto correlation function is described and optimal conditions of its registration are studied. First results following clinical application of the method are reported.

[Analysis of the structure of cardiac rhythm during treatment of auricular fibrillation with digitalis]. / Analiz struktury serdechnogo ritma pri digitalisnoi terapii mertsatel'noi aritmii.

The structure of cardiac rhythm was studied with the help of specialized computers during digitalization in 60 patients with atrial fibrillation of different etiology. The reduction of the heart contractions rate under digitalization is accompanied by certain changes in the structure of cardiac rhythm: the arrhythmic pattern of the ventricular contractions increases with a predominant growth of the number of long R--R intervals, the transitions from short intervals to longer ones become sharper, and portions of a relatively stable ventricular rhythm appear. Three main types of dynamics of the rhythm structure were distinguished on the basis of the changes in the interval R--R histogram. The described changes in the structure of cardiac rhythm are not specific, but when Digitalis drugs are used alone they can be used as additional criteria for digitalization.