DENDRITIC CELLS IN GLIOBLASTOMA TREATMENT: A MODERN VIEW OF THE PROBLEM AND OWN EXPERIENCE.

Glioblastoma (GBM) is the most aggressive primary malignant brain tumor in adults. The improvement of the efficacy of GBM treatment is an urgent problem encouraging the development of novel therapeutic strategies, in particular, immunotherapeutic modalities. With more understanding of the intimate interrelationships between the immune system and the mechanisms involved in cancer origin and progression, the skepticism related to the relevance of the immunotherapeutic approaches in the treatment of brain tumors is gradually decreasing.

RADIOSURGICAL TREATMENT OF RECURRENT GLIOBLASTOMA AND PROGNOSTIC FACTORS AFFECTING TREATMENT OUTCOMES.

Background: Glioblastoma (GBM) is the most prevalent malignant tumor of the brain in adults with the inherent aggressive behavior and high recurrence rate. The stereotactic radiosurgery (SRS) is currently considered as one of the effective modalities for GBM treatment allowing for the improvement of survival with the acceptable toxicity level.

Aim: To assess the effects of various factors on the survival of GBM patients following SRS.

Temozolomide in glioblastoma treatment: 15-year clinical experience and analysis of its efficacy.

Aim: To analyze retrospectively the efficacy of temozolomide (TMZ) in various treatment regimens in glioblastoma patients accounting for varying parameters of their treatment.

Materials And Methods: 302 glioblastoma patients were treated at the State Institution "Romodanov Institute of Neurosurgery of the National Academy of Medical Sciences of Ukraine" from 2003 through 2017. All the patients were surgically treated.

[Comparative effects of menthol and icilin on the induced contraction of the smooth muscles of the vas deferens of normal and castrated rats].

Non-specific TRPM8 agonist menthol was shown to inhibit voltage- and agonist-evoked contractions of the smooth muscle (SM) of rat vas deferens. Here we compared the action of menthol with the action of more specific TRPM8 agonist icilin on depolarization- (60 mM KCl), carbachol-(CCh) and noradrenalin-(Nor)-evoked contractions of the SM strips from the prostatic and epididymal portions of the vas deferens of normal and castrated (60-137 days) rats. Inhibitory action of menthol (100 microM) and icilin (10 microM) on the amplitude of KCl-, CCh- and Nor-induced contractions of normal as well as castrated rats was similar consisting about 50%, despite castration per se strongly potentiated CCh- and Nor-evoked contractions compared to the control animals.

[Changes in calcium-dependent potassium channels of isolated smooth muscle cells of the bladder in rats with experimental diabetes].

Activation of large conductance Ca2+-dependent potassium channels (BK channels) influences repolarization of the action potential and the level of the resting potential of detrusor smooth muscle cells (SMC). Overactive bladder syndrome (OBS) is one of the complications of diabetes. Here using whole-cell patch clamp technique we show sizable reduction of depolarization-evoked BK current (lBK) and decrease in the amplitude and frequency of spontaneous transient outward currents (STOCs) in isolated SMC from detrusor of rats with streptozocin-induced diabetes compared to control animals.

[Effects of N-stearoyl- and N-oleoylethanolamine on cardiac voltage-dependent sodium channels].

The group of N-acylethanolamines (NAE) includes lipids that are capable of modulating plasma membrane ion channels without involvement of cannabinoid receptors. However, the action of various members of NAE on voltage-gated Na+ channels (VGSC) in cardiac tissue is still not fully elucidated. Here using patch-clamp technique we have studied the modulation of biophysical properties of VGSC of neonatal cardiomyocytes by saturated N-stearoylethanolamine (NSE) and monounsaturated N-oleoylethanolamine (OEA).

[Urothelium-dependent modulation of urinary bladder smooth muscle contractions by menthol].

TRPM8 cold receptor/channel is considered amongst the variety of receptors that support and modulate sensory function of urothelium, although the information regarding this is still quite contradictory. Here we have studied the effects of nonspecific TRPM8 activator menthol on the contractions of the smooth muscle strips of the rat bladder with intact and removed urothelium, and assessed the expression in them of TRPM8 mRNA using semi-quantitative RT-PCR. Menthol (100 microM) decreased the basal tone and the amplitude of spontaneous contractions only in the strips with intact urothelium.

[Expression of the cold receptor TRPM8 in the smooth muscles of the seminal ejaculatory ducts in rats].

By means of real-time RT-PCR and immunofluorscent staining combined with confocal microscopy we show for the first time the expression ofmRNA and protein ofthe cold/menthol-sensitive cationic channel, TRPM8, in the smooth muscle cells (SMC) from the epididimal and prostatic portions of the rat vas deferens. Expression of TRPM8 mRNA correlated with the expression ofmRNA for androgen receptor (AR): orchidectomy of the animals resulted in the enhancement of the expression of both mRNAs, which likely reflects specific for the vas deferens compensatory response to the decreasing levels of circulating androgens. TRPM8 protein in the SMC from both parts of the vas deferens primarily localized outside the plasma membrane (PM), however, in the SMC from prostatic portion we observed higher TRPM8 protein targeting specifically the endoplasmic reticulum and PM, where it can form functional cold/menthol-sensitive cationic channel capable of modulating contractile activity of the smooth muscle.

[Gender differences in cold sensitivity: role of hormonal regulation of TRPM8 channel].

We have studied the gender differences in the perception of cutaneous cold stimuli in the innocuous temperature range (5-30 degrees C) in mice and rats. In the behavioral tests using two variable temperature plates technique female subjects displayed lower threshold for the sensation of cooling temperatures in the range of 15-25 degrees C compared to males. Patch-clamp experiments carried out on dorsal root ganglion (DRG) neurons from male and female rats maintained in the short-term cultures in the presence of testosterone or 17beta-estradiol, respectively, have revealed gender- and hormone-related differences in the electrophysiological properties of cold/menthol-sensitive TRPM8 channel: average density of menthol-activated I(TRPM8) current density in females' DRG neurons was higher compared to males', and the steady-state voltage-dependent activation curve of TRPM8 in females was shifted towards hyperpolarized potentials compared to males.

[Regulation of the excitability of neonatal cardiomyocytes by N-stearoyl- and N-oleoyl-ethanolamines].

N-acylethanolamines (NAE) are biologically active lipids able of modulating ion transport through the cellular plasma membrane, however specific targets of their action and signalling mechanisms involved in cardiac tissue are still poorly understood. Physiological activity of NAEs is known to depend on the level of unsaturation. Therefore, here we investigated the effects of saturated N-stearylethanolamine (NSE) and monounsaturated N-oleylethanolamine on electric excitability of neonatal rat cardiomyocytes.

[Modulation of the smooth muscle contractions of the rat vas deferens by TRPM8 channel agonist menthol].

TRPM8 is nonselective, Ca2- permeable cationic channel, which is activated by innocuous cold and by chemical drugs imitators of cooling, menthol, icilin and cucalyptol. TRPM8 expression was detected in the smooth muscle cells of the rat vas deference with preferential localization of the TRPM8 protein to the membrane of sarcoplasmic reticulum (SR). In the present work we have studied the effects of TRPM8 channel agonist, menthol, on the contractions of the smooth muscle strips of the epididimal and prostatic portions of the rat vas deferens evoked by potassium rich (KCl) Krebs solution and by muscarinic or adrenergic agonists carbachol (CCh) or noradrenalin (Nor).

[A hyperpolarization-activated current in rat menthol-sensitive sensory neurons].

In the present study we have investigated the correlation between hyperpolarization-activated current (1(h)) and menthol-activated current (I(TRPM8)) in rat dorsal root ganglion (DRG) neurons. We showed that I(h) is present in 89% of menthol-sensitive neurons which makes its presence reliable, though not absolute, criterion for pre-selection of such neurons. Endogenous I(h) recorded from different neurons exhibited variable density and activation kinetics.

[Functional identification of the TRPM8 cold receptor in rat prostate epithelial cells].

For the first time by means of electrophysiology we demonstrate functional expression of cold/menthol sensitive TRPM8 channel in secretory epithelial cells plasma membrane from rat prostate. Under whole-cell patch-clamp technique rapid cooling from 33 to 21 degrees C of freshly isolated rat prostate epithelial cells elicited the significant outward rectifying membrane current with close to 0 mV reversal potential characteristic of TRPM8 activation. Extracellular application of chemical analog of cooling, menthol (100 microM) at 33 degrees C evoked similar current responses.

[Calcium signaling in carcinogenesis].

Malignant transformation of the cells in cancer is caused by excessive proliferation accompanied by diminished ability for apoptosis due to the loss of normal control on these processes. Despite their ubiquity calcium ions are central to both processes, serving as major signalling agents. The present review examines what changes in calcium distribution among various compartments: extracellular space, cytoplasm, endoplasmic reticulum and mitochondria take place during transition from normal to pathological cell growth and death, what Ca2+-handling proteins are involved, and how this affects spatial and temporal characteristics of Ca2+ signals aimed at regulating specific cellular responses.

[Identification of E-4031-sensitive potassium current component in murine P19 embryonic carcinoma cell line differentiated in cardiomyocytes].

Pluripotent mouse P19 embryonic carcinoma cells represent a convenient in vitro model for studying various aspects of cardiac differentiation. Here by using whole-cell patch-clamp recording we have identified the rapid delayed rectifier K+ current, I(Kr) in P19 cell induced to differentiate into cardiac phenotype by DMSO (1%). Cardiac differentiation was confirmed by the appearance of spontaneously beating cells, their morphological features, ultrastructural clusterization of mitochondria around contraction elements, expression of cardiac actin mRNAs and MLC2v, and by the presence of inward sodium and calcium currents.

[Alterations in ATP-dependence of swelling-activated Cl- current associated with neuroendocrine differentiation of LNCaP human prostate cancer epithelial cells].

Increasing population of malignant, apoptosis resistant neuroendocrine (NE) cells due to differentiation of prostate epithelial/basal cells is a hallmark of advanced, androgen-independent prostate cancer, for which there is no successful therapy. Acquisition of apoptosis resistance involves alterations in the mechanisms of cell volume homeostasis, of which volume-regulate anion channels (VRAC) that carry swelling-activated Cl- current (I(Cl,swell)) represent one of the key determinants. Given that VRAC function is generally known to be ATP-dependent, here we investigated how such dependence may evolve during NE differentiation of LNCaP prostate cancer epithelial cells.

[Adenosine triphosphate-dependence of volume sensitive chloride current in LNCaP cell line of human prostate cancer].

Although sensitivity to intracellular ATP is considered to be one of the hallmarks of swelling activated Cl- current (I(Cl,swell)) involved in regulatory volume decrease (RVD) following hypotonic stress, the type and manner of such sensitivity seems to vary in different cell types. Here by using whole-cell patch-clamp recording we investigated ATP sensitivity of I(Cl,swell) in LNCaP human prostate cancer cell line. Suppression of endogenous ATP production with metabolic inhibitors (oligomycin, iodoacetate and rotenone) during cell dialysis with ATP- and Mg2+-free pipette solution did not prevent I(Cl,swell) in response to hypotonic exposure.

Incorporation of labelled N-acylethanolamine (NAE) into rat brain regions in vivo and adaptive properties of saturated NAE under x-ray irradiation.

Regional distribution of exogenous N-palmitoylethanolamine in the rat brain was investigated in the study. Possible protective and adaptive effect of N-stearoylethanolamine under 2 Gy whole-body X-irradiation and changes of brain lipid composition were also studied. It was found that after per os administration to rats N-([9,10-3H]-palmitoyl)-ethanolamine was primarily accumulated in hypothalamus, pituitary and adrenal glands and the label amount in brain was 0.

[Calcium-dependent programmed cell death in prostate cancer].

In the present review we describe the major molecular determinants of calcium homeostasis in prostate cancer cells and establish their role in the transformation to apoptosis-resistant cell phenotypes typical of advanced androgen-independent prostate cancer. We show that the hallmark of such transformation is complete loss of apoptosis pathway associated with endoplasmic reticulum calcium store depletion.

[Effect of neuroendocrine cell differentiation of the human prostatic neoplasm cell line LNCaP on the characteristics of volume-sensitive chloride content].

By means of patch-clamp technique we examined changes in volume-regulated chloride current (ICl,swell) at neuroendocrine differentiation of androgen-dependent LNCaP prostate cancer cells. In those cells with neuroendocrine differentiation resulted from an increase in the intracellular cAMP, ICl,swell became much faster in response to applying external hypotonic solution and cell swelling. Changes in final rectification and voltage-dependent inactivation were not detected, as compared to the control cells.

[The influence of extracellular pH on volume-activated chloride current in the prostate cancer epithelial cells].

The influence of extracellular pH on characteristics of volume-activated chloride current, ICl,swell, in the human prostate cancer epithelial cell line, LNCaP, was studied using the patch-clamp technique. Acidification of the extracellular hypotonic solution used to develop the current shortened the temporal parameters of ICl,swell development and reduced its maximal density. Sudden shifts of the pH towards acidification caused fast, transient potentiation of the current followed by its sustained inhibition.

[Testosterone modulation of HERG potassium channel blockade induced by neuroleptics].

The repolarisation phase of cardiac action potential is characterized by sexual dimorphism suggesting the role of sex steroid hormones in the regulation of K+ channels. Here we report on the effect of testosterone on blockade of HERG-encoded K+ channels induced by neuroleptics. These compounds are used in clinics to treat psychiatric disorders, but reportedly have proarrhythmic side effects, on HERG-encoded K+ channels responsible for the rapid component of cardiac delayed rectifier K+ current, IKr.

[Effects of sex hormones on HERG potassium channels expressed in Xenopus oocytes].

The repolarisation phase of cardiac action potential is characterized by sexual dimorphism suggesting the role of sex steroid hormones in the regulation of K+ channels. Here we report on the effects of testosterone and 17 beta-estradiol on HERG-encoded K+ channels, expressed in Xenopus oocytes. At 1M concentration testosterone decreased the amplitude of HERG-directed IKr (rapid component of cardiac delayed rectifier K+ current) by 30% within 30 min of exposure, while 17-estradiol had no effect.

[Blockade of HERG K+ channels expressed in Xenopus oocytes by antipsychotic agents].

We have investigated the effects of neuroleptic agents, haloperidol, pimozide and fluspirilen, that are used in clinics to treat psychiatric disorders, but reportedly have proarrhythmic side effects, on HERG-encoded K+ channels responsible for the rapid component of cardiac delayed rectifier K+ current, IKr. All three agents blocked HERG-directed IKr in Xenopus oocytes in a voltage-dependent manner. The extent of the blockade increased with depolarization correlating with channels activation consistent with open-channel blocking mechanism.