Biocompatible photocrosslinked poly(ester anhydride) based on functionalized poly(epsilon-caprolactone) prepolymer shows surface erosion controlled drug release in vitro and in vivo.

Star-shaped poly(epsilon-caprolactone) oligomers functionalized with succinic anhydride were used as prepolymers to prepare photocrosslinked poly(ester anhydride) to evaluate their in vivo drug delivery functionality and biocompatibility. Thus, in this work, erosion, drug release and safety of the photocrosslinked poly(ester anhydride) were examined in vitro and in vivo. A small water-soluble drug, propranolol HCl (M(w) 296 g/mol, solubility 50 mg/ml), was used as the model drug in an evaluation of the erosion controlled release. Drug-free and drug-loaded (10-60% w/w) poly(ester anhydride) discoids eroded in vitro (pH 7.4 buffer, +37 degrees C) linearly within 24-48 h. A strong correlation between the polymer erosion and the linear drug release in vitro was observed, indicating that the release had been controlled by the erosion of the polymer. Similarly, in vivo studies (s.c. implantation of discoids in rats) indicated that surface erosion controlled drug release from the discoids (drug loading 40% w/w). Oligomers did not decrease cell viability in vitro and the implanted discoids (s.c., rats) did not evoke any cytokine activity in vivo. In summary, surface erosion controlled drug release and the safety of photocrosslinked poly(ester anhydride) were demonstrated in this study.

In vivo delivery of a peptide, ghrelin antagonist, with mesoporous silicon microparticles.

Peptides may represent potential treatment options for many severe illnesses. However, they need an effective delivery system to overcome rapid degradation after their administration. One possible way to prolong peptide action is to use particulate drug delivery systems. In the present study, thermally hydrocarbonized mesoporous silicon (THCPSi) microparticles (38-53 microm) were studied as a peptide delivery system in vivo. D-lys-GHRP6 (ghrelin antagonist, GhA) was used as a model peptide. The effects of GhA-loaded THCPSi microparticles on food intake (s.c., GhA dose 14 mg/kg) and on blood pressure (s.c., GhA dose 4 mg/kg) were examined in mice and rats, respectively. In addition, the effects of THCPSi microparticles (2 mg) on cytokine secretion in mice after single s.c. administration were examined by determining several cytokine plasma concentrations. The present results demonstrate that GhA can be loaded into THCPSi microparticles with a high loading degree (20% w/w). GhA loaded THCPSi microparticles inhibited food intake for a prolonged time, and increased blood pressure more slowly than encountered with a GhA solution. Furthermore, THCPSi microparticles did not increase cytokine activity. The present results suggest that THCPSi might be used as a drug delivery system for peptides.

Proteins and peptides of the salivary gland secretion of medicinal leeches Hirudo verbana, H. medicinalis, and H. orientalis.

The protein and peptide composition of medicinal leech salivary gland secretion (SGS) was analyzed in preparations obtained in July from three species--Hirudo verbana, H. medicinalis, and H. orientalis. Two-dimensional electrophoresis (molecular mass 10-150 kD and pI 3-10) revealed no distinctions in the distribution of over 100 silver-stained proteins. Differences were noted only in intensity of 10 protein spots at 30-90 kD and pI 4.7-7.5. Mass spectrometric profiling of SGS of the three leech species using the Zip-Tip/golden chip scheme and cation-exchanging chips CM-10 revealed over 50 components in SGS of each of the three leech species. It was noted that 30-40% of the individual masses of the SGS of each leech species fall within the masses present in SGS of at least one of the two other species. This rather small part of the total mass may be indicative of a high polymorphism of amino acid sequences or a high frequency of posttranslational modifications of the SGS proteins and peptides. Calculation of Jacquard's coefficient showed that H. medicinalis and H. orientalis are closest to each other in SGS composition, which is consistent with data in the literature on the phylogenetic relationship between these two species of medicinal leech. Comparison of detected molecular masses with those of six known biologically active compounds produced by medicinal leeches revealed their uneven distribution in SGS of each of the three medicinal leech species. This opens prospects for using certain species of medicinal leech for targeted therapy of various pathologies.

Mass spectrometric profile of the serum as a marker of experimental psychoemotional stress in rats.

Mass spectrometric profiling of blood serum from stressed and control rats was used for the detection of stress marker proteins. Profiling with reversed-phase chips revealed 6 discriminatory mass spectrometry peaks. Cluster analysis showed that the animals can be divided into 2 groups by the intensity of these peaks. One group mainly consists of stressed rats, while other includes control specimens. Profiling of serum samples after fractionation on reversed-phase magnetic granules allowed us to identify an 8910-Da biomarker. Our results indicate that mass spectrometric profiling holds much promise for the detection of stress markers in the serum.

Molecular interactions of acute phase serum amyloid A: possible involvement in carcinogenesis.

Acute phase serum amyloid A (A-SAA) is a well-known marker of inflammation. The present review summarizes data on the regulation of A-SAA expression, signaling pathways which it is involved in, its effects, and possible influences on progression of malignant tumors.

Two-dimensional electrophoretic proteome study of serum thermostable fraction from patients with various tumor conditions.

One of the problems of plasma proteomics is a presence of large major components. In this work, we use the thermostable fraction as a way to deplete these major proteins. The thermostable fraction of serum samples from patients with ovarian, uterus, and breast cancers and benign ovarian tumor was analyzed using two-dimensional electrophoresis combined with MALDI-TOF(-TOF)-mass spectrometry. Of them, alpha-1-acid glycoprotein and clusterin are expressly down-regulated in breast cancer, whereas transthyretin is decreased specifically in ovarian cancer. Apolipoprotein A-I forms have decreased spot volumes, while haptoglobin alpha1, in contrast, is elevated in several tumors. These data are partly consistent with previous art studies on cancer proteomics, which involve mass-spectrometry-based serum profiling techniques. Serum thermostable fraction may be recommended as a good tool for medium and small protein proteome investigation, in particular, by 2D-electrophoresis.

Mechanism of adaptation of the vascular system to chronic changes in nitric oxide level in the organism.

We studied the possibility of directed modulation of the efficiency of NO storage in rats due to adaptation to the chronic changes in plasma NO level. The efficiency of NO storage increased during long-term maintenance of high plasma level of NO and decreased in NO-deficient states. The compensatory changes in NO storage capacity of vessels depending on its organism content represent a new mechanism of adaptation of the cardiovascular system to chronic excess or deficit of NO, while directed modulation of this process can be important for the protection of the organism against both surplus or shortage of NO.

Adaptation to hypoxia prevents disturbances in cerebral blood flow during neurodegenerative process.

The rats with neurodegenerative brain disorder induced by administration of a toxic fragment of beta-amyloid demonstrate weakened endothelium-dependent dilation of cerebral vessels, which attested to impaired production of endothelial NO. At the same time, toxic beta-amyloid fragment induced the formation of NO depots in the walls of cerebral vessels, which indirectly attests to NO overproduction in the brain tissue. Preadaptation to hypoxia prevented endothelial dysfunction and improved the efficiency of NO storage. Our results suggest that adaptation to hypoxia protects the brain from various changes in NO production during neurodegenerative damage.

[Molecular diagnostics of ovarian cancer using proteome techniques].

Ovarian cancer is most common gynecological malignancy which is difficult for early diagnostics. Development of new methods for diagnostics of ovarian cancer, especially, on early stage, is the urgent problem of modern oncology. A general approach to early diagnostics of cancer is a discovery of specific blood disease biomarkers. Ovarian cancer biomarker used in the modern art, CA125, has some drawbacks resulting in extensive research recently directed to this tumor diagnosis. In particular, it was shown the advantage of parallel use of several diagnostic biomarkers instead of the single one. Proteome techniques (two-dimensional electrophoresis, mass-spectrometry methods) in connection to bioinformatics represent a powerful tool for new biomarker discovery. In this respect, the best method of choice is shown to be SELDI-TOF (Surface-enhanced laser desorption/ionization time-of-flight) technique which combines chromatography protein chip application and mass-spectrometry-based detection. In this review, new ovarian biomarker data obtained by proteome methods are summarized.

Detection and description of various stores of nitric oxide store in vascular wall.

We analyzed the possibility of the existence of various NO pools in the vascular wall. Incubation of isolated rat aorta with dinitrosyl iron complex (NO donor) led to the formation of NO stores in the vascular wall detected by vascular relaxation response induced by diethyldithiocarbamate and N-acetylcysteine. Comparison of the effects of successive application of diethyldithiocarbamate and N-acetylcysteine revealed two NO pools (one pool responded to both agents, while other responded only to N-acetylcysteine). Inhibition of guanylate cyclase with methylene blue abolished the response to diethyldithiocarbamate, while the reaction to N-acetylcysteine decreased by the value, corresponding to diethyldithiocarbamate-dependent relaxation. It is hypothesized that in the vascular wall NO is stored in the form protein-bound dinitrosyl iron complexes and S-nitrosothiols in hydrophilic and hydrophobic cell compartments.

Effect of transmural pressure on constrictor reactions of caudal artery in hypotensive and hypertensive rats.

Clipping of the abdominal aorta distally to the renal arteries produces a persistent decrease in blood pressure in hindquarter vessels by 35-40%. On week 6-7 postoperation, the reactions of the caudal artery perfused in vitro under constant pressure to norepinephrine were studied. At transmural pressure of 150 mm Hg, the vascular responses in hypotensive rats were reduced compared to those in normotensive control. By contrast, the responses of hypertensive vessels were more pronounced at 75 mm Hg even after deendothelization.

[Prenatal diagnosis of cytomegalovirus infection]. / Prenatal'naia diagnostika tsitomegalovirusnoi infektsii.

Prognostic value of markers of cytomegalovirus infection (CMV) in pregnant women for the neonatal status was assessed. Detection of such markers as antiCMV IgM and CMV DNA in cervical secretion by DNA dot-spot hybridization in women with a complicated course of pregnancy indicates a 5.7% risk of delivery of children with stable symptoms. Studies of antibodies to pre-early proteins (IE CMV) showed that antiCMV IgG to IE are more incident in pregnant women than antiCMV IgM; moreover, antiCMV IgG to IE but not antiCMV IgM are detected in umbilical blood. The results of detection of antiCMV IgG and IgM to IE correlated with the clinical characteristics of newborns.

Wall thickness and constrictive responses of the caudal artery in rats with renovascular hypertension.

In rats with hypertension modeled by the one kidney-one clamp method, constrictory responses of the isolated caudal artery to norepinephrine differed under various perfusion conditions. Vascular reactions in hypertensive rats were more potent at a constant flow rate, and less potent at a constant pressure compared to those in normotensive rats. Previous experiments demonstrated similar changes in constrictory responses of the caudal artery in spontaneously hypertensive rats. It is assumed that these peculiarities of the vascular reactivity during hypertension are determined by thickening of the smooth muscle layer of the vascular wall.

Responses to noradrenaline of tail arteries in hypertensive, hypotensive and normotensive rats under different regimens of perfusion: role of the myogenic response.

The vasoconstrictor effects of noradrenaline were studied in spontaneously hypertensive rats (SHR) compared with Wistar Kyoto rats (WKY), and in Wistar rats with regional hypotension (WH) compared to control Wistar rats (WC). The abdominal aorta was ligated in WH distal to the renal arteries, lowering blood pressure in the hindquarters by 41% and tail artery wall cross-sectional area by 35% compared with WC. A cylindrical segment was dissected from the proximal part of the tail artery, cannulated at both ends and perfused with Krebs-Henseleit solution either at constant flow starting from a pressure of 120 mmHg or at a constant pressure of 120 mmHg. The cumulative dose-response relationships for noradrenaline were determined in control conditions and subsequently in the presence of gadolinium (100 microM), a non-specific blocker of mechanosensitive channels. Under constant-flow perfusion noradrenaline evoked a more prominent resistance increase in SHR compared with WKY and in WC compared with WH. Similar relations were seen in the presence of gadolinium, although responses were reduced. At constant pressure perfusion the vasoconstrictor response to noradrenaline was lower in SHR compared with WKY and in WC compared with WH. Application of gadolinium under constant-pressure perfusion reduced responses in WKY and WH, so that vasoconstriction in SHR became more pronounced than that in WKY and in WC than that in WH. It is suggested that the results can be explained by the difference in wall thickness causing different degrees of activation of the myogenic response to distension.

[The effect of symptom-free genital herpes on the course and outcome of pregnancy]. / Vliianie bessimptomnogo genital'nogo gerpesa na techenie i iskhody beremennosti.

The incidence of primary genital herpes among women of the childbearing age in Khabarovsk is 2.6% or from 0 to 10.4 per 1000. Asymptomatic herpes simplex virus (HSV) carrier state in the genital tract was detected in 24.1 +/- 5.2% normal women. The virus-carrier state exerts untoward effects on the course of pregnancy and condition of the fetus. In women infected with HSV, the hazard of abortion is 1.8 times as frequent, premature delivery is 3.2 times as frequent, and children of such mothers have symptoms of intrauterine hypotrophy 4.5 times as frequently as normally.

[A comparison of the results of analyzing the clinical data from gynecological, ophthalmological and dental patients using a set of rapid methods for the diagnosis of herpesvirus infection]. / Sopostavlenie rezul'tatov analiza klinicheskikh materialov ginekologicheskogo, oftal'mologicheskogo i stomatologicheskogo profilei naborom ékspress-metodov diagnostiki gerpesvirusnoi infektsii.

Analysis of biological specimens from gynecological, ophthalmological, and stomatological patients was carried out using current express methods for diagnosis of herpes virus infection developed by the authors: enzyme immunoassay (EIA), time-resolved fluoroimmunoassay (tr-FIA), molecular hybridization and conventional immunofluorescence technique. Monoclonal antibodies (MCA) generated by us with epitope specificity for gpD HSV-2 were used in EIA and tr-FIA. F(ab')2-fragments of MCA used as sorbents and for preparation of conjugates were found to be by 250% more sensitive than intact IgG used for similar purposes. The employment of EIA based on MCA F(ab')2-fragments contributed to detection of 0.2-0.6 ng of virus-specific protein in a specimen. Comparison of the results of analysis of clinical specimens of broad spectrum (mucous secreta from the cervical canal of the uterus and posterior vault of the vagina, saliva, lacrimal fluid) by the above express methods for diagnosis of herpes virus infection revealed a high percentage of positive findings, none of the methods providing 100% diagnosis. The use of a set of methods makes the resultant information more objective. Therefore, in future the data obtained by a number of current express methods for herpes diagnosis should be compared.

[The use of an immunoglobulin in the comprehensive treatment of ophthalmic herpes]. / Primenenie immunoglobulina v kompleksnom lechenii oftal'mogerpesa.

The paper analyses results after treatment of 30 patients with herpetic keratitis by means of a specific antiherpetic immunoglobulin manufactured at the Sverdlovsk Research Institute of Virus Infections. As compared with traditional treatment, the terms of clinical recovery and the number of bed-days reduced. The preparation can be used in a complex treatment of patients with a herpetic disease.