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Pompe disease (PD) is a rare autosomal recessive muscle lysosomal glycogenosis caused by a deficiency of acid-α-glucosidase. There are two main forms of the disease: aggressive infantile PD started within the first year of life with a severe enzyme deficiency and multiorgan involvement, and late onset PD (LOPD) with progressive signs and symptoms including predominant proximal, axial muscle weakness and respiratory insufficiency started at any time from 1 till 75 years and older. Usually due to physician's unawareness, most adults with PD are diagnosed with great delay. The typical features and early nonspecific signs in four patients, aged between 35 and 72 years, with confirmed LOPD are delineated and discussed in correspondence with the age of first signs, age development of muscle weakness, distribution and age of final diagnosis. The disorders for differential diagnosis and spectrum of conditions that expanded the possibility of PB are listed. The fluorometrically analyzed level of acid α-glucosidase from dried blood spots is considered to be the first choice diagnostic method for clinically suspected cases of LOPD.

Based on the measurements of fluorescence of bark chloroplasts by means of PAM and PEA fluorometers, the information capacity of the methods for assessing the physiological state of Tilia cordata L. from the maximum quantum efficiency of PS II photochemistry (Fv/Fm) and the performance index (PI) has been compared. The measurements were performed on annual shoots of linden trees growing in different environment. It was shown that the chlorophyll content in the bark of shoots growing near the busy urban street was twice less compared with trees growing out of the city. On the trees from the unsafe environment, a small decrease in the relative fluorescence variable (Fv/Fm) was registered, and there was a significant statistical deviation of this value compared to control trees. It was found that the PI and its constituent parameters calculated on the basis of light fluorescence induction curve (PEA-method) are more informative and allow one to recognize changes in the primary energy transformation processes in PS II when they are comparatively small. The results of our work show that PI can be used as a sensitive and a rapid test to evaluate the physiological state of trees and other plant objects even under minor environmental changes.

It has been shown by fluorescence analysis in vitro that the water-soluble sodium salt of the polycarboxylic derivative of fullerene C60, fullerenol, and complexes of fullerene C60 with polyvinylpyrrolidone (mol. wt. 25000 and 10000) destroy amyloid fibrils of the brain peptide Abeta(1-42) and prevent their formation. The results of fluorescence analysis confirmed the data obtained earlier by high-resolution electron microscopy. Fluorescence analysis and electron microscopy are complementary methods for the selection of effective antiamylod drugs.

The development of malignant neoplasm of the head and neck is related to the state of intracellular proteasome system. Elevation of total activity of proteasomes and specific activity of 20S proteasomal pool are accompanied by changes in proteasomal composition. Tumor size correlated with the content of regulatory proteasomal complex PA28. In the presence of regional metastases, 26S proteasome activity decreases and the content of proteasome immune subunit LMP7 in the tumor increases.

The current investigation was undertaken with the aim to carry out an in vitro evaluation of the ability of coumarin derivatives as probe substrates to predict the activity of CYP51b1. The results obtained indicate that 7-aminocoumarin-4-acetic acid (ACAC) can be used to determine the recombinant CYP51b1 activity. Determination of CYP51b1 activity with ACAC is based on the direct registration of fluorescence increasing at 30 degrees C. It was found also that BMR in a simple soluble model system can be used as an electron donor for CYP51B1. Fluorescence-based assay is highly sensitive and can be used for the screening of sterol 14alpha-demethylase inhibitors.

5'-Pyrenylmethylphosphoramidite and 5'-bispyrenylmethylphosphordiamidite derivatives of oligo(2'-O-methylribonucleotides) and their analogues with thymidine attached at their 3'-termini by a 3'-3'-phosphodiester internucleotide bond (inverted thymidine) were synthesized. The effect of the pyrene residue(s) on the thermal stability of duplexes of the modified oligonucleotides with RNA and DNA was studied. A possibility of detection of hybridization of 5'-mono- and 5'-bispyrenyl derivatives with RNA and DNA targets in solution was demonstrated according to the changes in fluorescence. 5'-Pyrenylmethylphosphoramidite derivatives of oligo(2'-O-methylribonucleotides) and their inverted analogues were shown to serve as sensitive probes for the detection of oligonucleotide substitutions in RNA and DNA by the method of thermal denaturation of the formed duplexes detected according to changes in their fluorescence.

The distribution in nature and the spectral and structural properties of chromoproteins of the green fluorescent protein (GFP) family and their differences from one another and other fluorescent proteins of this family are considered. Discussed in detail are practical applications of the chromoproteins and their mutant variants that have unique characteristics not found among natural proteins of the GFP family, such as far-red or photoconvertible fluorescence, a large Stokes shift, enhanced phototoxicity, etc.

Simvaglyzin, a complex compound of simvastatin and glycyrrhizic acid, administered to rabbits with experimental hypercholesterolemiain in doses equivalent to 66.6 and 40 microg/kg simvastatin exhibited antioxidant capacity (decreased the content of lipid peroxidation products in the blood by 27-41%) and endothelium-normalizing effect (decreased the level of von Willebrand factor and endothelin-1 by 26-58 and 21-29%, respectively, compared to 200 microg/kg simvastatin, p<0.05).

Using the screening model of hypochromism, we showed that caffeine forms regular clusters consisting of 8-12 molecules. Addition of 7-aminoactinomycin D (7AAMD, a fluorescent analogue of actinomycin D) to the clusters leads to its sorption on the cluster surface. Photoexcitation of 7AAMD leads to its desorption from the surface into the aqueous phase and emission of a quantum. Fluorescence of 7AAMD in the presence of caffeine clusters is quenched by dinitrophenol more weakly than without clusters (the quenching constants are approximately 85 and approximately 280 M(-1), respectively) due to decreased steric availability of the antibiotic to the quencher. Addition of 7AAMD-caffeine complexes to DNA leads to a long-wavelength shift in the excitation spectrum and an increase in the fluorescence intensity along with a shift of the fluorescence spectrum to the short-wavelength area. This fact reflects redistribution of the antibiotic from the caffeine surface to the hydrophobic areas inside DNA. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2008, vol. 34, no. 2; see also http://www.maik.ru.

The purpose of the present study was to detect the main histamine-containing structures of the rat uterus and to study the changes of its content during the estrous cycle. The work was performed during the autumn-winter period using 120 female rats that were divided into groups according to the stages of the estrous cycle: early estrus, late estrus, metestrus, early diestrus, late diestrus, proestrus. Cryostat sections of uterine corpus and cervix were studied using fluorescent-histochemical method with the orthophthalic aldehyde. The histamine content was determined in mast cells, macrophages, surface and glandular epithelial cells, smooth myocytes in the tunics of the uterine corpus and cervix, as well as in the contents of the uterine cavity. During the sexual cycle, the highest histamine concentrations were detected in mast cells, especially in those located in the myometrium. In most of the bioamine-positive cellular elements of the uterine wall, the minimal histamine content was found in metestrus. As a result of a computer-statistical analysis, it was demonstrated that the intraorgan histamine metabolism in the structures of the uterine tunics studied, was characterized by phase and regional specificity together with the dynamic nature of temporal coupling of histamine content.

The fluorometric method was used to study guantitative parameters of phagocytosis of fluorescein-labeled Escherichia coli cells by mouse peritoneal macrophages. E. coli cells were conjugated with fluoresceinisothiocyanate (FITC) and then incubated with macrophages. At the end of the assay phagocytosis was arrested with a lysing solution (0.5% Triton X-100 in 0.01 M phosphate-buffered 0.15 M saline, pH 7.4). Trypan blue at a concentration of 0.04% was used as a quenching agent to differentiate between attachment and ingestion of E. coli cells. The time course analysis within this method showed that phagocytosis of E. coli cells was temperature and opsonin dependent. The number of E. coli cells ingested by macrophages increased rapidly during the initial 60 min of incubation at 37 degrees C. E. coli cells required opsonization with 5% native serum to achieve their optimal uptake. The uptake of nonopsonized bacteria by macrophages was significantly lower that that of opsonized ones (P < 0.05). It was demonstrated that sodium azide inhibited phagocytosis of E. coli cells by mouse peritoneal macrophages in a dose-dependent manner.

The results of studying the genotoxic effect of radiation on endometrial cells of irradiated rats using comet assay correlated with the index of spontaneous DNA degradation. Studying the genotoxic effect of in vitro irradiation (2 Gy) on human lymphocytes by means of chromosome analysis and fluorometric assay of nucleoids DNA in irradiated leukocytes also yielded similar results. Measuring of the biochemical marker holds much promise as an express method of evaluation of the genotoxic effect of radiation on mammalian somatic cells.

The alkylhydroxybenzene (AHB) autoregulatory factors d1 (fd1) of microorganisms have been found to directly interact with highly polymeric DNA. This circumstance results in changes, related to alterations in the topology of this macromolecule, in DNA physicochemical properties. The physicochemical properties of DNA in the presence of chemical analogues of microbial AHBs (methylresorcinol; hexylresorcinol; and 2-(4-hydroxyphenyl)ethane-1-ol, also known as tyrosol) were investigated using adsorption spectrophotometry, fluorometry, heat denaturation, viscosimetry, and electrophoresis in agarose gel. A number of concordant effects pointing to DNA-AHB interactions were revealed that manifesed themselves in the hypochromic properties of the resulting complexes, an increase in their melting temperature and viscosity, a decrease in their electrophoretic mobility, and a change in the fluorescent properties of AHBs upon complexation with DNA. Such alterations were particularly significant in the presence of hexylresorcinol, which possessed the maximum alkyl radical length among the fd1 analogues tested. Using atomic force microscopy, we visualized the micelle-like DNA nanostructures forming in the presence of AHBs. The results obtained provided the basis for developing a hypothetical model of the interaction between the biopolymer macromolecule and low-molecular-weight AHBs that takes into account the differences in the hydrophobicity of individual AHB homologues functioning as ligands. In terms of our model, we discuss AHB involvement in the stabilization of DNA and alteration of its topology, i.e., in the process related to intragenomic rearrangements, which account for the intrapopulational variability of bacteria, including dissociation processes.

A technique for the fluorometric determination of protein and non-protein thiols in brain tissue with the maleimide reagent ThioGlo-1is has been described. Comparison with the most popular spectrophotometric Ellman method has been performed. The ThioGlo-1-based method is highly sensitive and may be used for thiol determination both in homogenates and supernatants of brain tissue.

The paper represents data on new potentialities in evaluating a severity of patients with total peritonitis (TP). They are based on the fluorescence analysis of the physical-and-chemical properties of blood albumin. The findings are a result of a multi-center research undertaken by clinics of Moscow and Moscow Region. The properties of albumin-molecule binding sites were investigated by the fluorescence test. The parameter, measured in the test, was named "effective albumin concentration". The study protocol comprised the data of clinical examinations, scales SAPS and MPI as well as laboratory findings and EAC. The subsequent analysis denoted that a combination of 2 parameters, i.e. EAC and serum urea concentration routinely measured, is highly informative in evaluating the TP severity and correlates well with the survival rate in the discussed pathology. The study results could be a basis for an early individual prognostication of TP course made on the 1st postoperative day.

The concentration and properties of the blood-albumin linkage centers were examined by the method fluorescent probes, besides, the general clinical-and-laboratory indices in the blood serum of patients with diffusive purulent peritonitis were determined. The parameters as obtained by the method of fluorescent probes revealed a valuable information on a patients' severity condition and on a forecasted disease evolution; finally, they surpass, in a majority of cases, the generally accepted clinical-and-laboratory tests applicable to similar conditions.

We used rat liver tissue slices and isolated hepatocytes to demonstrate that single exposures to a "weak" carcinogen 4-dimethylaminoazobenzol (DAB) or to a "strong" one, such as N-diethylnitrosamine (DENA), cause identical antigenic restructuring to occur. It presented as formation of membranous heteroorganic antigens of renal origin on hepatocyte surfaces. Antigens were associated with Zajdela's hepatoma; their synthesis lasted longer after exposure to DENA. They, however, were not identified in intact rat liver. Their synthesis, following single exposure, was assayed immunocytofluorimetrically. It is suggested that the antigens be used as markers of cell dysdifferentiation and malignancy and testing substances for carcinogenicity.

A new approach to detecting induced mutations was tested based on the assay of cell extracts and special growth media following cultivation of UV irradiated Escherichia coli cells. No correlation was found between the UV dose and the optical densities of cultural media or cell extracts prepared by Triton X-100 treatment. Blue fluorescence of concentrated cultural media varied with cell dose, according to a rather complex law, which differed substantially from the known dose-effect curves for induced mutations. Nevertheless, a certain extent of the brown staining of tryptophan containing medium could, presumably, serve as a quite sensitive indicator of the integral metabolic activity of bacteria grown in the medium. Besides, we observed that overnight lag phase cultures became gradually more transparent, when analysed in the spectrophotometer cuvette just after their dilution with fresh medium.

A total of 147 patients with preproliferative and proliferative diabetic retinopathy (DR), with the exudative hemorrhagic and nonexudative stages (93 females and 54 males), were consulted at Helmholtz Institute of Ophthalmic Diseases in 1997-2001. Control group consisted of 39 healthy subjects aged 25-76 years. The group of patients with preproliferative DR consisted of 27 patients with compensated diabetes mellitus. The course of diabetes in 39 patients with proliferative DR was evaluated as medium severe during the subcompensation stage. The clinical picture of the fundus oculi was characterized by pronounced hemorrhagic activity. Slight retinal hemorrhages were seen in the patients with preproliferative DR. Nodules, defects of pigmented epithelium, atrophic foci were seen in the central zone of the fundus oculi in patients with nonexudative stage of central chorioretinal dystrophy; edema in the central zone, polymorphic hemorrhages, solid exudate were observed in the patients with the exudative hemorrhagic stage; subretinal neovascular membrane was detected in some patients. Erythrocyte deformability coefficient, platelet aggregation coefficient to ADP, and platelet factor were evaluated by common methods. Serotonin was measured by the fluorometric method (B. M. Kogan's method) at clinical biochemical laboratory of urgent methods of examination of N. V. Sklifosovsky Institute of Emergency. The erythrocyte deformability coefficient was notably increased in the patients with proliferative DR and central chorioretinal dystrophy in comparison with the normal value. Plasma serotonin concentration was increased significantly only in the patients with proliferative DR, while in the rest groups this concentration was notably decreased. Study of platelet aggregation gave contradictory results. The values of platelet factor 4 differed from the control negligibly. Serotonin insufficiency in patients with proliferative DR was paralleled by increased plasma serotonin concentration and thrombocytopathy. In patients with preproliferative DR and central chorioretinal dystrophy serotonin insufficiency was associated with decreased concentration of serotonin and thrombocytopathy. Erythrocyte rigidity was similarly increased in patients with proliferative DR and central chorioretinal dystrophy.

The circadian rhythm of grazing of microalgae by a laboratory culture of Ceriodaphnia quadrangula under continuous illumination was studied by continuous registration of chlorophyll fluorescence at the outlet of a flow-through cultivator. A culture of green alga Chlorella vulgaris was used as a feed. The data obtained were treated by the statistical spectral analysis. It was found that animals preliminarily grown under a 12 h light: 12 h dark regime and transferred to constant light showed two maxima in the circadian rhythm of grazing with periods of 0.7 and 1.1 h. Animals preliminarily grown under constant light showed no circadian rhythm of grazing. It was concluded that the circadian rhythm of grazing of C. quadrangula has endogenous nature and can change according to light conditions.

Serum albumin parametres in the rats with various prognostic resistance against emotional stress in the control and in conditions of experimental emotional stress were investigated. Analysis of action of peptides raising the animals' resistance against emotional stress (DSIP and SEMAX) in rats with different prognostic resistance against emotional stress, on the serum albumin characteristics, was carried out.

It was shown that the addition of synthetic six-membered peptide (HLDF-6) and its Tyr-analog (HLDF-Y) to cultural medium significantly increased the survival of cells HL-60, treated by cold shock. The prophylactic administration of HDLF-Y (1 mg/kg, 4 hours prior to applied actions) decreased the response of hypothalamushypophysis-adrenal glands system and sympathicoadrenal system of rat males on supercooling and also increased the resistance of mouse males to supercooling and X-irradiation. In the experiences with females HDLF-Y did not show the similar biological activity.

An influence of middle-wave ultraviolet radiation (lambda max = 306 nm) on plasma membranes of mice peritoneal macrophages was studied by microfluorimetry analysis. It was found that a percentage of cells with damaged plasma membranes in the irradiated macrophage population reliably increased with the UVB dose starting with 6 J/cm2. Irradiation of cells with 4.2 J/cm2 UVR dose which does not cause evident damage to plasma membranes led to the latent damage which was detected by treatment with detergent digitonin (4.5 micrograms/ml).

The influence of hypothermal stress (+4 degrees during 3 h) on the ways of serotonin metabolism in pineal gland and its structure has been studied in dynamics on adult male Wistar rats. It has been revealed that melatonin-producing epiphyseal function suffers from phase changes in dynamics of adaptation--significant rising during 15 min. after beginning of the experiment, rehabilitation up to normal--in 30 min, and fast suppressing--in 3 hrs. Suppressing of the functional pineal activity is not due to switched serotonin metabolism with melatonin and new indoles release, but to a partial pinealocytes breaking from their active function.

The provision of children from Baikal Lake areas with selenium was studied in relation to the geochemical area, the man-made pollution of an area, sex, age, and the presence of a disease has been studied. The content of selenium in the hairs of the children living in the areas extensively exposed to pollution is show to be lower than in those from ecologically pure areas. The correlation of selenium with other elements in the children's hair depends on the level of these elements. Thus, with low hair zinc content, there is a positive correlation (r = 0.32) between selenium and zinc (p < 0.05), while with its hair levels, there is a negative correlation (r = -0.46; p < 0.05). The content of selenium in the children's biosubstrata increases with age. The hair selenium levels are higher in girls than in boys. In ill children, they amount to 47 to 77% of those of healthy children.

The effect of urea (1-3 M) on conformational changes in the active center of native and reconstituted Ca-ATPase of sarcoplasmic reticulum modified by fluorescein-5-isothiocyanate (FITC) was studied using the method of fluorescence titration by neodymium (Nd3+) ions. Based on the analysis of curves of fluorescence quenching of FITC-labeled Ca-ATPase by Nd3+ ions, the parameters characterizing the structural changes in the Mg-ATP binding center were determined. It was assumed that FITC and Nd3+ ion bind to different polypeptide fragments moving relative to each other, which provides the conformational lability of the nucleotide binding site at some stages of the catalytic cycle. A comparison of structural changes caused by urea at the active site of native and reconstituted Ca-ATPase of sarcoplasmic reticulum indicates that the Nd3+ binding center is localized in the region of contacts of monomers in the oligomer.

The structural and functional characteristics of bean leaves (the content of chlorophyll, the rate of oxygen production, the slow fluorescence induction, and light-induced changes in the EPR signal I from oxidized reaction centers P700+) were investigated to obtain insight into the mechanism of influence of zinc chloride on the photosynthetic apparatus. Seedlings were grown on hydroponic medium containing ZnCl2 at concentrations from 10(-7) to 10(-3) M. At low concentrations of ZnCl2, a decrease in the content of chlorophyll per one unit of leaf mass was observed, while the rate of oxygen production per chlorophyll was increased. High concentrations of ZnCl2 in the hydroponic medium caused the slowed down the plant development and inhibited the light-induced production of oxygen. The changes in biophysical characteristics of leaves the parameter FM/FT of the slow fluorescence induction, and kinetics of redox transients of P700 induced by ZnCl2 were of similar character and correlated with the changes in photosynthetic activity. The data obtained demonstrate that structural and functional changes in the photosynthetic apparatus induced by the variations of growth conditions have adaptive character.

The effect of selective agents on the efficiency of Agrobacterium-mediated transformation of pear was shown. The transformation frequency of the pear stock PS no. 217 by a binary vector carrying the nptII gene conferring kanamycin resistance was 0.4 or 3.1%, while the hpt gene for hygromycin resistance used as a selective marker increased transformation frequency to 6.2 (11.5%). In addition, upon selection on hygromycin B, the proportion of pseudotransgenic regenerants considerably decreased. In four transformation experiments, twenty independent clones were recovered, and their transgenic nature was confirmed by PCR, histochemical, and fluorometric analyses of GUS activity. The presence of introns in the coding region of a heterologous gene was shown to influence the efficiency and stability of transgene expression in plant tissues. Fluorometric determination of GUS activity conducted for a period of two years demonstrated a threefold increase in transgene expression in the case that an intron-containing construct was used for transformation. The expression level was rather stable across several years. The transformation procedure developed may be used for successful expression of heterologous genes controlling agronomic characters in pear plants.

Influence of dietary supplement Tinrostim-C on cholinesterase (ChE) activity and serum lipids peroxidation (LP) in patients whose work connects with emotional stress was examined. Activity of ChE was measured by Ellman calorimetric method (with acetylthiocholin as substrate), LP--by fluorimetric method with malone dialdehyde. Tinrostim-C was given three times a day in 0.5 g. On the 10th day of taking the preparation an activity of serum ChE increased 23.5% higher and had been staying higher during the whole period of observation. In vitro experiments showed an activating effect of Tinrostim-C and piracetam for serum ChE. The level of LP being initially higher was decreasing to values close to normal and had been staying at decreased level during the whole period of observation.

The comparative study incubation influence at 37 degrees C on the lymphocyte Fc gamma-receptor (Fc gamma R) binding ability to IgG1 and IgG2 and on the antibody-dependent cell mediated cytotoxicity (ADCC) was performed. By means of the fluorometric binding assay it was determined that during 30 min of incubation the binding ability of Fc gamma R decreased while after 180 min of incubation it increased. Upon comparing both IgG1 and IgG2 binding abilities to lymphocytes no statistically important changes were noticed (p < 0.05). The changes in the binding ability of Fc gamma R for IgG1 and IgG2 had a similar pattern to the changes.

Fluorometry was done in 52 patients with diabetes mellitus to determine the blood plasma content of free histamine and free serotonin. The control group was 29 essentially healthy subjects. Significant increase was found out in free histamine during the stage of decompensation of diabetes mellitus (P < 0.01). With compensation of diabetes mellitus setting in, the content of histamine returns to normal (P < 0.01). The blood plasma level of free histamine can be used as an additional criterion in the assessment of compensation of diabetes mellitus. Free histamine is supposed to play an active part in the pathogenesis of diabetic angiopathies.

Aim: To investigate composition of very low density lipoproteins (VLDLP) and some subfractions of low density lipoproteins (LDLP), and their resistance to oxidation in patients with non-insulin-dependent diabetes mellitus (NIDDM) varying in severity compared to healthy subjects., Materials and Methods: 26 NIDDM patients and 16 healthy controls. VLDLP and subfractions of LDLP were isolated by ultracentrifugation. The lipoproteins were assayed for protein, cholesterol, initial products of lipid peroxidation (LPO). After autooxidation of lipoproteins, fluorimetry examined the degree of the oxidant modification., Results: The initial level of LPO products was elevated in VLDLP and light subfractions of LDLP in severe and moderate NIDDM patients. Even in mild NIDDM, resistance to oxidation of dense LDLP subfractions was low and continued to decrease with aggravation of the disease. High ratio cholesterol/ protein occurred in VLDLP and subfractions of LDLP from patient with severe NIDDM., Conclusion: It is shown that subnormal resistance to oxidation of LDLP dense subfractions even in mild NIDDM is an informative diagnostic criterion of risk factor to develop atherosclerosis.

Bacterial sensitivity to different various phages was examined by electro-orientation spectroscopy, fluorometry, and electron microscopy. The strains of Pseudomonas aeruginosa, Staphylococcus aureus, Yersinia pestis, Mycobacterium smegmatis, and Xanthomonas campestris were used. The fluorescence intensity of a membranotropic agent in the ANS-cell-phage system was shown to depend on the interaction of a bacterial virus and a microorganism. Fluorometric data correlated with electro-orientation spectroscopic findings. An analysis of the low-frequency site makes it possible to determine phage adsorption on the bacterial surface. The changes in electro-orientation effects at high frequencies suggest that there are barrier dysfunctions in the external membranes and that there is cellular phage reproductions. Whether fluorometry and electro-orientation spectroscopy can be further used for rapid identification of microorganisms by using phages is discussed.

Investigation to effect of low radiation doses in the T-cells activation surface markers expression of resident in 4 radiation zone around Chernobyl. Was observed the increase cells level, which CD4+CD25+ and CD4+HLA-DR(+)-cells markers expression. Were revealed the increase of the apoptosis related CD4+CD+ cells level in peripheral blood.

According to electron-microscopic data, various cells in the M. smegmatis ATCC607 population interact differently with phage MTPH11. Fluorometric studies of phage-host interactions were performed using a membranotropic fluorescent probe, 8-anilino-1-naphthalene sulfonate (ANS). Changes in the electric characteristics of mycobacterial cells infected with the phage were studied by electro-orientational (EO) spectroscopy. The problem of the employment of fluorometry and EO spectroscopy for rapid phage typing of mycobacteria is discussed.

The most popular object for studying endocytosis of EGF-receptor complexes, human epidermoid carcinoma A431, was shown to answer to EGF in high concentration (100 ng/ml) by growth inhibition, being indifferent to lower (0.1-1 ng/ml) concentrations. At the same time, cells NIH 3T3, expressing human EGF receptor (HER14), and epithelial mammary cells HC11 increased 14C-thymidine incorporation into DNA after EGF addition. However, for HER14 cells stimulatory effect of EGF was twice weaker than that induced by serum, whereas the effect of EGF on 14C-thymidine incorporation in DNA of cells HC11 was approximately 5 times stronger compared to serum. Therefore, cells HC11 may be referred to as EGF-dependent. Cell cycle analysis by fluorimetry showed that more than 90% of serum-starved HER14 and HC11 were in G0/G1. Within 19-20 h after stimulation by EGF 70-90% of HC11 cells and only 30-40% of HER14 cells were in S-phase. EGF removing from culture medium earlier than 9-11 h after stimulation blocked entering of HC11 cells into S-phase, whereas such EGF-dependent period was not found for cells HER14. Thus, synchronization of progression through early stages of cell cycle, stimulated by EGF and the presence of well defined "early" (EGF-dependent) and "late" (EGF-independent) phases, make cells HC11 convenient object for studying physiological role of EGF receptor complexes endocytosis.

FcR on chicken blood lymphocytes have been studied by fluorimetric binding assay. Binding data were analyzed by Scatchard's method. Temperature and time variations were investigated in order to optimise the binding conditions. It was concluded that binding FcR to IgG on chicken blood lymphocytes was optimal at 0 degree C for 60 min. 10-14% of nonspecific binding were detected in different experiments. Scatchard binding plots at 0 degree C demonstrated that IgG bound to FcR per cell 2.3 +/- 4 x 10(5) with affinity 3.9 +/- 0.5 x 10(5) M-1.

Transcapillary metabolism of the retina and effects of triglycerides on the transcapillary function of the retina in diabetes mellitus were examined in 79 patients (mean age 45.4 +/- 3 years) with various stages of diabetic retinopathy (DR) by fluorometry of the vitreous body. The filtration in the blood-retina direction was 1.2-1.3 times higher than in the control as early as during the functional and preclinical stages of DR, and the coefficient of fluorescein withdrawal from the vitreous body was 4.5 to 6.2% decreased. As DR progressed, the permeability of the blood-retinal barrier in the blood-retina direction increased and the discharge from the retina to the blood was decelerated, which was confirmed by a progressive prolongation of diffusion of 0.1 ml of 1% fluorescein solution injected subconjunctivally: from 1.3 +/- 0.2 and 1.4 +/- 0.5 h in the functional and preclinical stages, respectively, to 2.7 +/- 0.3 h in the proliferative stage of DR versus 1.05 +/- 0.01 h in the control. The level of serum triglycerides and coefficients of fluorescein entry in and withdrawal from the vitreous body, time of papule diffusion, and vascular changes in the microcirculatory vessels reliably correlated.