Enhanced Proapoptotic Effects of Water Dispersed Complexes of 4-Thiazolidinone-Based Chemotherapeutics with a PEG-Containing Polymeric Nanocarrier.

AIM: To study whether water formulation of the complex of 4-thiazolidinone derivatives with a PEG-containing polymeric nanocarrier enhances their pro-apoptotic action towards rat glioma C6 cells. METHODS: Mechanisms of antineoplastic effects of 4-thiazolidinone derivatives were investigated in vitro with rat glioma C6 cells. Cell nativity, cell cycling pattern, and Annexin V expression were evaluated and DNA damage was estimated by DNA comet analysis. A novel water-based formulation of 4-thiazolidinone derivatives complexed with a polymeric nanocarrier was utilized for enhancing pro-apoptotic action towards C6 cells. RESULTS: The studied 4-thiazolidinone derivatives use apoptosis mechanisms for killing rat glioma C6 cells, as confirmed by FACS analysis of these cells in pre-G1 stage, the appearance of Annexin V positive C6 cells, and an increased number of DNA comets of higher classes. Complexation of the studied compounds with a PEG-containing polymeric nanocarrier significantly increased pro-apoptotic effects in rat glioma C6 cells measured by all methods mentioned above. CONCLUSION: Complexation of 4-thiazolidinone derivatives with a PEG-containing polymeric nanocarrier provided them with water solubility and enhanced pro-apoptotic effects in rat glioma C6 cells.

Enhanced anticancer activity and circumvention of resistance mechanisms by novel polymeric/ phospholipidic nanocarriers of doxorubicin.

Severe toxic side effects and drug resistance are the major limitations of doxorubicin (Dox), one of the most potent anticancer agents in clinical use. Nanocarrier preparations offer the opportunity to overcome these drawbacks, which is reflected in the clinical approval of two liposomal Dox preparations. Additionally, there are many attempts to enhance the activity of Dox against multi-drug resistant (MDR) cancer cells. However, most of these strategies resulted in the increased uptake of Dox in resistant cells, only, while it remained unchanged in chemo-sensitive cells. Here, we present a new polymeric-phospholipidic hybrid delivery system which distinctly enhanced the accumulation and activity of Dox in all tested cancer cell lines including several MDR cell models. Notably, the resistance levels against Dox were reduced from about 6-fold to about 2-fold. Moreover, the new nanocarriers were shown to rapidly (within 10 min) and effectively transport Dox into resistant as well as sensitive cancer cells. Consequently, treatment with the new Dox-containing nanocarriers resulted in effective cell cycle arrest in G2/M phase and ROS-induced cell death induction. Finally, the new nanocarriers were tested against NK/Ly lymphoma and L1210 leukemia cells in vivo. In both cell models, the nanoformulation of Dox resulted in 100% cured animals already at low concentrations (0.1 mg/kg), while free Dox solely extended survival time. This indicates that the incorporation of phospholipids into PEGylated polymeric nanocarriers is a promising strategy to enhance efficacy and reduce toxicity of Dox treatment against both sensitive and resistant cancer models in vitro and in vivo.

Nanoformulation improves activity of the (pre)clinical anticancer ruthenium complex KP1019.

Ruthenium anticancer drugs belong to the most promising non-platinum anticancer metal compounds in clinical evaluation. However, although the clinical results are promising regarding both activity and very low adverse effects, the clinical application is currently hampered by the limited solubility and stability of the drug in aqueous solution. Here, we present a new nanoparticle formulation based on polymer-based micelles loaded with the anticancer lead ruthenium compound KP1019. Nanoprepared KP1019 was characterised by enhanced stability in aqueous solutions. Moreover, the nanoparticle formulation facilitated cellular accumulation of KP1019 (determined by ICP-MS measurements) resulting in significantly lowered IC50 values. With regard to the mode of action, increased cell cycle arrest in G2/M phase (PI-staining), DNA damage (Comet assay) as well as enhanced levels of apoptotic cell death (caspase 7 and PARP cleavage) were found in HCT116 cells treated with the new nanoformulation of KP1019. Summarizing, we present for the first time evidence that nanoformulation is a feasible strategy for improving the stability as well as activity of experimental anticancer ruthenium compounds.

[Study of rat blood serum biochemical indicators of cardiotoxic action of novel antitumor 4-thiazolidinone derivatives and doxorubicin in complexes with polyethylene glycol-containing polymeric carrier in the rat blood serum].

The aim of this study was to measure the activity of enzymes which reflect cardiotoxic action in rats of novel synthetic 4-thiazolidone derivatives--3882, 3288 and 3833 that demonstrated antineoplastic effect in vitro towards 60 lines of human tumor cells tested in the framework of the program of screening new anticancer drugs at the National Cancer Institute (USA). Such action of these compounds was compared with the effect of well known anticancer agent doxorubicin and after conjugation of all above mentioned substances with new polyethylenglycol-containing polymeric comb-like carrier that was synthesized by the authors. Among the biochemical indicators of cardiotoxic action of anticancer agents, activity of the following enzymes in rat blood serum showed to be the most informative: creatine kinase, lactate dehydrogenase, aspartate aminotransferase, and alanine aminotransterase. Tenfold injection of doxorubicin in a dose of 5.5 mg/kg of weight caused rats' death, while 3882, 3288 and 3833 preparations had not such action. Application of the doxorubicin in combination with polymeric carrier prolonged the survival time to 20 days. Thus, the injection of anticancer agents in a complex with polymeric carrier provides a significant decrease in their cardiotoxicity that was confirmed by the corresponding changes in the activity of marker enzymes: creatine kinase, lactate dehydrogenase, aspartate aminotransferase and alanine aminotransferase in blood serum of treated rats.

[Effect of free and polymer carrier encapsulated doxorubicin towards HCT116 cells of human colorectal carcinoma].

Development of novel nanoscale functionalized carriers is nowadays one of the most urgent problems in cancer treatment. The aim of our study was to compare the antineoplastic effect of free doxorubicin and its complex with a nanoscale polymeric carrier towards HTC116 colorectal carcinoma cells. It was established that application of the complex of poly(5-tret-butylperoxy)-5-methyl-1-hexene-3-in-co-glycydyl metacrylat)-graft-polyethyleneglycol (poly(VEP-GMA-PEG)-graft-PEG), where VEP--5-tret-butylperoxy)-5-methyl-1-hexene-3-in; GMA--glycydyl metacrylat; graft-PEG--graft-polyethyleneglycol accordingly, functionalized with phosphatidylcholine for doxorubicin delivery increased 10 times the efficiency of cytotoxic action of this drug, as compared wich such efficiency in case of the action of free doxorubicin. The encapsulated form of doxorubicin caused more intensive cleavage of the reparation enzyme PARP and longer delay in G2/M cell cycle arrest, compared to such effects of free doxorubicin. The developed carrier itself is non-toxic to the used mammalian cells and does not cause impairment in their cell cycle. A deletion in both alleles of p53 gene did not affect the antineoplastic action of doxorubicin that was immobilized on the nanoscale carrier. Thus, p53-dependent signaling pathways are not involved in the cytotoxic action of doxorubicin-carrier complex. It is suggested that novel nanoscale polymeric carrier poly(VEP-GMA-PEG)-graft-PEG functionalized with phosphatidylcholine could be a promising carrier for targeted delivery of anticancer drugs.

[Antitoxic and antioxidant effects of N-stearoylethanolamin in the content of nanocomposite complex with doxorubicin in organs of mice with Lewis carcinoma].

The aim of the study was to evaluate the possibility to reduce the doxorubicin toxic effects by its immobilization with N-stearoylethanolamine (NSE) on nanocarier polyethylene glycol. The studied parameters of the doxorubicin toxicity were: the level of creatinine in the mice blood plasma and activity of alanine aminotransferase and aspartate aminotransferase in the blood plasma of mice. The activity of catalase superoxide dismutase, glutathione peroxidase and intensity of lipid peroxidation was determined in the tissues of the heart, kidneys and liver. Doxorubicin in the content of nanocarrier alone caused an increase of serum creatinine and aspartateaminotrasferase activity in plasma of experimental animals with carcinoma. Nanocomposite which contained doxorubicin and NSE, did not cause an increase of these parameters. It has been shown that the administration of a carrier containing doxorubicin to mice with Lewis lung carcinoma caused the decrease of catalase activity in mice with carcinoma. The combination of NSE and doxorubicin on the carrier led to the normalization of this parameter to the level of intact animals. NSE immobilized on a carrier together with doxorubicin caused a decrease in the activity of superoxide dismutase in the kidney tissue of mice with tumor. The tumor growth caused the increase of the of superoxide dismutase in mice. The administration of a carrier which contained doxorubicin and NSE normalized superoxide dismutase in heart tissue contrary of kidney. The obtained results show the antitoxic and antioxidant effects of N-stearoylethanolamine immobilized in the nanocarrier complex together with doxorubicin.

[Protective effect of N-stearoylethanolamine in suspension and in nanocomposite complex in the organs of mice with the Lewis carcinoma under doxorubicin intoxication administration].

The antioxidant effects of N-stearoylethanolamine (NSE) in the nanocomplex composition and in suspension are shown on the model of intoxication by doxorubicin in conditions of development of the Lewis carcinoma in the heart, kidneys and liver tissue and in the blood plasma of female mice. The NSE suspension reduces the level of urea in the blood plasma of mice with the Lewis carcinoma, which growth was revealed as a result of introduction of doxorubicin. Under introduction of nanocomplex the amount of urea remains at the level of that in the intact mice. In the blood plasma of mice with the Lewis carcinoma the NSE suspension and nanocomplex reduce activity of aspartate aminotransferase, the basic marker of necrosis of the heart tissue, growth of which was caused by the tumour development. Doxorubicinum increases activity of alanine aminotransferase, the marker of the liver lesion; introduction of NSE in the nanocomplex composition prevents the growth of the enzyme activity. N-stearoylethanolamine, both in the nanocomplex and in suspension, modulates activity of enzymes of antioxidantive protection of the heart, kidney and liver tissue of mice with the Lewis carcinoma.

[Morpho-physiological features of the human eye sclera as the key unit in the level of intraocular pressure under normal conditions and in glaucoma].

The results of sclera investigations from the positions of morphology, physiology, biomechanics and control-system theory, are summarized. The morphological evidence was obtained supporting the physiological hypothesis stating that the specific anatomical organization and spatial displacement of elastic fiber-containing scleral layers against one another, are the key elements in formation of the individual level of intraocular pressure (IOP) in man both under normal and pathological conditions. It was shown that the elastic fibers outlined the collagen lamallae in of scleral internal layers. External scleral layer lacked elastic fibers. Scleral elastic fibers are necessary for the functioning of the mechanism of sclera microfluctuations associated with the intraocular fluid accumulation and removal, they are also important for dampening the sudden changes of IOP. Under normal conditions, age-related increase in scleral rigidity is primarily associated with the process of accelerated aging of its superficial non-elastic layers, resulting in the physiological response of the current IOP level elevation. As IOP becomes elevated under normal conditions, the internal elastic fiber-containing scleral layers are increasingly pressed against more rigid external layer. This limits the displacement capacity of internal layers against each other, resulting in the decline of the efficiency of elastic fiber work in dampening the sudden changes of IOP. In the healthy eyes, the process of scleral aging brings to a natural development of ophthalmohypertension, when IOP elevation is physiologically required for the maintenance of the volume microfluctuation mechanism ("scleral respiration"). In glaucoma, the pathological rearrangement of the scleral fibrous structures is observed, resulting in an additional abrupt increase of its rigidity, with the reciprocal significant elevation of the current level of IOP and the amplitude of its jumps. Pathophysiological mechanism of these significant changes in glaucoma remains currently unknown, however, morphological evidence indicates that it is associated with the changes in the metabolic processes in sclera.

[Ultrasonic frequency currents and laser radiation in combined treatment of glaucoma].

A total of 178 patients aged 53 to 78 years with advanced and progressive primary open angle glaucoma have undergone a hypotensive operation. It is shown that it is beneficial for the patients to combine medication with ultratonotherapy of eyelid region for restoration of hydro- and hemodynamics in the operated eye; to give an impact on the projection of the cortical sight centers for activation of trophic and neuroregulatory mechanisms of visual functions. Such a combined approach improves the course of the postoperative period and prolongs remission in glaucomatous atrophy of the visual nerve.

[EHF-therapy of herpetic keratouveitis]. / KVCh-terapiia gerpeticheskikh keratouveitov.

A device BIK for active suppression of infection has been used against Herpes virus infection type 1 and 2 in 72 patients with keratouveitis. The immunological and genotypic monitoring show that patients with herpetic keratouveitis after BIK treatment have lower titers of antiherpetic antibodies, normal cellular and humoral immunity, weaker viremia and reduced quantity of viral material in corneal tissue. This positive effect persisted for 8-12 months. Thus, BIK device provides a definite, prolonged and stable therapeutic effect in patients with herpetic keratouveitis.

[Combined treatment of congenital blepharoptosis in children and adolescents]. / Kombinirovannoe lechenie vrozhdennogo blefaroptoza u detei i podrotskov.

The paper deals with a complex procedure for rehabilitation of children with congenital blepharoptosis, which includes the multilevelled impact on the concerned central and peripheral links of the neuromuscular apparatus by using magnetic laser therapy and electrostimulation.

[Electrostimulation and magnetic therapy in the treatment of accommodation cramp and in the prevention of progressive myopia in children and adolescents]. / Elektrostimuliatsiia i magnitoterapiia v lechenii spazma akkomodatsii i v profilaktike progressirovaniia miopii u detei i podrostkov.

The paper presents a procedure and results of use of electrostimulation, magnetic therapy, and electrophoresis during treatment for accommodation cramp and in the prevention of myopia in children and adolescents. Data that characterize the dynamics and stability of achieved results are given.

[Use of transconjunctival electroimpulse therapy in diseases of the optic nerve]. / Priminenie metoda transkoniunktivalnoi elektroimpulsnoi terapii pri zabolevaniiakh zritelnogo nerva.

Seventy-one patients with optic nerve abnormalities of different origin were treated using transconjunctival electrostimulation of the eyeball with electrodes positioned in the ciliary body projection. A positive effect was attained, particularly manifest in the acute period of optic nerve diseases (neuritis, anterior ischemic neuropathy) and in involvement of the optic nerve in glaucoma patients. The results were assessed from the principal parameters of visual function, intraocular pressure, data of hydrodynamic and electrophysiological studies.

[Surface tension of diluted solutions of human tears in the normal state and in various types of eye pathology]. / Poverkhnostnoe natiazhenie razbavlennykh rastvorov sleznoi zhidkosti cheloveka v norme i pri razlichnykh patologiiakh glaza.

It is shown that the measurement of diluted solutions surface tension of normal tears and tears at different eye pathologies (cataract, glaucoma before and after operation) gave identical results. It is interpreted as a normal function of the lacrimal gland at these pathologies.

[The role of myeloperoxidase, a product of the polymorphonuclear leukocytes, in the pathogenesis of cataract]. / Rol' mieloperoksidazy--produkt polimorfoiadernykh leikotsitov v patogeneze katarakty.

The capacity of myeloperoxidase which is the product of polymorphonuclear leucocytes to induce the lens opacity was studied in young and old rabbits. It was found that the injection of myeloperoxidase solution into anterior chamber of the eye causes the irreversible lens opacification in old rabbits, not in young ones. Light microscopy of the lens section has shown the following alterations: the local thickening of the anterior capsule, disorderly accumulation of epithelial cells, formation of so-called "bladder cells" under the lens epithelium. Changes in experimental eyes are typical for cataract.

[Immuno-correcting therapy of sympathetic ophthalmia]. / Immunokorrigiruiushchaia terapiia simpaticheskoi oftal'mii.

Exchange plasmapheresis was used in correction of the immune homeostasis in 5 patients with sympathetic ophthalmia. The inflammatory process ceased in all the cases. Exchange plasmapheresis resulted in improvement of the blood supply to ocular vessels, of intraocular vessel function, and of metabolic processes in the external layers of the retina. Cellular and humoral immunity parameters normalized. The hemo- and hydrodynamic parameters of the eye improved; all these results evidence that exchange plasmapheresis is an effective method in the treatment of sympathetic ophthalmia.