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The influence of amiodarone on intracellular transport of calcium ion in cardiomyocytes of rat was investigated. The experiments were performed on isolated papillary muscles of Wistar rats. Force-frequency dependence (0.7, 1, 2, 3, 4 Hz), extrasystolic and postextrasystolic contractions and post-rest (4-60 s) reactions of rat myocardium after amiodarone treatment (1 μM) were investigated. Decay potentiation coefficient of contraction force was estimated. Results. The analyses of force-frequency dependence has shown that amiodarone prevent the decreasing of the force contraction at increasing of the stimulation frequency. Amiodarone promotes increase of the time constant t1(T50), that indicate the drug promotes acceleration of Са2+ transport inside the SR resulting increase of Са2+ in the places of its release from the sarcoplasmic reticulum (SR). Treatment of papillary muscle with amiodarone decreased amplitude of extrasystolic contractions. As known, postextrasystolic and post-rest reactions of myocardium characterize the SR function. We have found amiodarone increased potentiation of postextrasystolic and post-rest contractions. Preliminary caffeine perfusion of muscles preparations cancelled the amiodarone-induced increasing postextrasystolic and post-rest potentiation. However, potentiation decay coefficient before and after treatment with amiodarone didn’t have difference. Conclusions, amiodarone influences on intracellular calcium ions homeostasis by modulation SR functions related with most likely are stipulated either by activation of Са2+ transport from uptake sites to release sites or by prevent of Са2+ leakage from the SR.

Change of interval–force dependence of rat papillary muscles at acute and course use of amiodarone was investigated. Methods. The experiments were performed on isolated papillary muscles of Wistar rats. The extrasystolic (0,2-1,5 s), postextrasystolic contractions and post-rest (4-60 s) reactions of rat left papillary muscles perfused Krebs-Henseleit solution with amiodarone (1 μM) and papillary muscles of rat treated amiodarone in dose 20 mg/kg/day for 14 days were investigated. Results. It has been found that both acute and course use of amiodarone decreases the amplitude of extrasystolic contraction. At the same time, only acute use of amiodarone leads to additional enhancement of potentialtion of contractile response at extrasystolic impulse and performance of post-rest test. Conclusion was made, that both acute and course use of amiodarone decreases excitability of cardiomyocytes but only at course use this antiarrythmic drug increases effective refractory period of myocardium. Acute exposure of amiodarone is accompanied with change of functional state of cardiomyocyte sarcoplasmatic reticulum.

Background: The aim of our study was to determine if ablation and pacing improved brain perfusion (BP) and cognitive function (CF) in patients with medically refractory rapidly conducted atrial fibrillation (Med Refr RCAF). Methods and Results: The study included 17 patients with Med Refr RCAF (average age 55.3 ± 4.5 years). All patients underwent brain single photon emission computed tomography scanning with 99mTc-hexamethylpropylene amine oxime and comprehensive neuropsychological testing before and after 3 months following pacemaker implantation. The BP was significantly lower in all regions in patients with Med Refr RCAF compared with the control group. The greatest BP decrease was revealed in the inferior frontal (P = 0.002) and posterior parietal (P = 0.024) brain regions. These patients showed cognitive deficit in 94%. There was a direct correlation between BP and CF parameters. Ablation followed by pacemaker implantation had a positive effect on BP and CF in all patients with Med Refr RCAF. Thus, BP increased in the right inferior frontal (P = 0.01), in the left superior frontal (P = 0.007), and in the left temporal (P = 0.005) cortex. These patients demonstrated improvements in immediate and delayed verbal memory, immediate and delayed visual memory, abstract mentation, attention, psychomotor speed, as well as in learning. Conclusions: Patients with atrial fibrillation and rapid ventricular rates refractory to medical treatment have marked signs of brain hypoperfusion and impaired CF. Ablation and pacing improve left ventricular systolic function, thereby increasing BP and improving CF. (PACE 2011;1–7)

It has been shown that pectin polysaccharides from different plants, depending on their structure, can either protect the intestinal walls of mammals against damage and inhibit the development of inflammation or, on the contrary, have proinflammatory effects. At the same time, galacturonan isolated from any pectin, being the main carbohydrate chain (backbone) of its macromolecule, shows a marked anti-inflammatory effect. A decrease in the quantity of neutrophiles in the intestinal wall after induced inflammation indicates that the anti-inflammatory effects of pectins can be based on their influence on the functional activity of leukocytes.

Pediatric glioblastoma (pGBM), although rare, is one of the leading causes of cancer-related deaths in children, with tumors essentially refractory to existing treatments. We have identified IGF1R to be a potential therapeutic target in pGBM due to gene amplification and high levels of IGF2 expression in some tumor samples, as well as constitutive receptor activation in pGBM cell lines. To evaluate the therapeutic potential of strategies targeting the receptor, we have carried out in vitro and in vivo preclinical studies using the specific IGF1R inhibitor NVP-AEW541. A modest inhibitory effect was seen in vitro, with GI50 values of 5 to 6 μmol/L, and concurrent inhibition of receptor phosphorylation. Specific targeting of IGF1R with short interfering RNA decreased cell viability, diminished downstream signaling through phosphoinositide 3-kinase (PI3K), and induced G1 arrest, effects mimicked by NVP-AEW541, both in the absence and presence of IGF2. Hallmarks of PI3K inhibition were observed after treatment with NVP-AEW541 by expression profiling and Western blot analysis. Phospho–receptor tyrosine kinase (RTK) arrays showed phosphorylation of platelet-derived growth factor receptor (PDGFR) α/β in pGBM cells, suggesting coactivation of an alternative RTK pathway. Treatment of KNS42 with the PDGFR inhibitor imatinib showed additional effects targeting the mitogen-activated protein kinase pathway, and cotreatment of the PDGFR inhibitor imatinib with NVP-AEW541 resulted in a highly synergistic interaction in vitro and increased efficacy after 14 days therapy in vivo compared with either agent alone. These data provide evidence that inhibition of IGF1R, in combination with other targeted agents, may be a useful and novel therapeutic strategy in pGBM. Mol Cancer Ther; 10(8); 1407–18. ©2011 AACR.

The results of investigations of zinc coatings obtained on the surface of metals in the process of mechanochemical synthesis realized in conditions of vibro-wave technological systems (VTS) are presented. The features of the morphology of the coating structure are revealed, which activates the role of indents in the formation of free-moving vibrations under the influence of low-frequency oscillations, and its influence on increasing the operational properties of the surface layer of the parts. The advantages of this method of applying zinc coatings in comparison with traditional methods are shown.

A pectic polysaccharide named capsicuman (CA) was isolated from fresh sweet pepper by extraction with a saline solution containing hydrochloric acid (pH 1.5) and pepsin at 37 °C for 4 h. Capsicuman was shown to consist of d -galacturonic acid (GalA, 74.0%), rhamnose (Rha, 1.6%), arabinose (Ara, 2.6%) and galactose (Gal, 2.4%) residues. This polysaccharide was digestible with 1,4-alpha- d -galacturonase to yield d -GalA, thus confirming capsicuman as a pectic polysaccharide. Partial acid hydrolysis of capsicuman revealed galacturonan to be the core of the macromolecule. Purified capsicuman (CA-2) was obtained from CA by ion-exchange chromatography on DEAE-cellulose. Nuclear magnetic resonance (NMR) spectra indicated that the backbone of capsicuman contained 1,4-alpha- d -galacturonan partially substituted with methyl and O-acetyl ester groups. After oral administration to mice, capsicuman CA, CA-2 and the galacturonanic fragment of CA (CA-H) were found to decrease tumour necrosis factor-alpha TNF-alpha release and to increase production of interleukin-10 (IL-10) in lipopolysaccharide (LPS)-stimulated whole blood. This pectin was also shown to improve the survival of mice that were subjected to a lethal dose of LPS. The present study demonstrates that the pectin capsicuman CA, which possesses anti-inflammatory properties, can be isolated from fresh sweet peppers using extraction with simulated gastric media.

It was shown that the energy metabolism of the heart mitochondria of experimental animals and patients is more resistant to damage at combined postinfarction cardiosclerosis and diabetes in comparison with the individual pathology. We found that the changes of free fatty acid content and conjugation of the processes of oxidation and phosphorylation in heart mitochondria are components of the metabolic stability of myocardium at the combined development of postinfarction cardiosclerosis and diabetes mellitus. Our data demonstrate a direct link between the violations of the processes of oxidative phosphorylation and accumulation of free fatty acids owing to change in activity of endogenous phospholipases, in particularly, mi- tochondrial phospholipase A2. Similar results were obtained for intraoperative biopsy specimens of patients’ hearts, and of adult Wistar rats’ hearts. We hypothesized that the preservation of energy metabolism is a manifestation of summing up of compensatory processes at de- velopment of nonspecific response of cells to damage at the early stages of pathological pro- cess.

We studied the effects of recombinant granulocytic CSF on heart remodeling in BALB/c mice after cryodestruction. Administration of granulocytic CSF was started 1 day after cryodestruction (subcutaneously, 10 μg/kg/day, for 4 days). As early as after the first injection, leukocytosis in the peripheral blood started to develop, leukocyte count peaked on days 4-6 and returned to normal on day 14. Treatment with granulocytic CSF significantly increased the content of progenitor cells in the bone marrow and led to rapid development of the inflammatory reaction and myocardium infiltration with mononuclear cells. Injections of granulocytic CSF did not reduce scar area, but provided significantly less pronounced heart hypertrophy, which attests to its better functional properties. By day 30 after cryodestruction, control animals and animals receiving granulocytic CSF exhibited similar morphological picture at the site of damage. Thus, our regimen of granulocytic CSF administration produced a mobilizing effect on bone marrow progenitor cells and postinfarction heart remodeling. Direct effects of granulocytic CSF on the heart have to be established for its use in the treatment of myocardial infarction.

Sea-level fluctuations in the terminal Eocene, Oligocene, and Neogene of the Eastern Paratethys are quantitatively assessed on the basis of facies and old coastlines traced on the northern platform shelf, levels of river valley incisions, and the study of seismic profiles. The first data massif allows the characterization and correlation of transgression stages in the history of the Eastern Paratethys. The greatest transgressions fall within the first half of the Late Eocene, mid-Early Oligocene, initial Late Oligocene, initial Early Miocene, the initial Tchokrakian, Karaganian and Sarmatian in the Middle Miocene, the middle and late Sarmatian and early Pontian in the Late Miocene, and the Akchagylian in the Caspian basin of the Pliocene. In contrast, the greatest incisions of northern rivers running from the platform allow us to establish the time and extent of the main declines in the base levels of the erosion. Maximal incisions date back to the terminal Eocene-initial Oligocene, terminal Solenovian time in the terminal Rupelian, the terminal Maikop in the Early Miocene, the terminal Sarmatian and middle Pontian in the Late Miocene, and the Early Pliocene in the Caspian basin. Large regressions also formed unconformity surfaces, traced on seismic profiles as erosion boundaries of several orders. The surfaces are confined to the Eocene/Oligocene boundary, middle and late Maikop, Sarmatian/Meotian boundary, middle Pontian, and terminal Miocene-initial Pliocene, as well as being traced even in the most deep-water basins. The synthesis of these data suggests a preliminary version for the curve of transgression-regression cyclicity. Its correlation with the eustatic curve shows their similarity only in the lower part-prior to the initial Middle Miocene, when Paratethys became a semi-closed basin.

A pectic polysaccharide called apiuman was isolated from fresh celery stalks by extraction with an aqueous ammonium oxalate followed by ultrafiltration, and was shown to consist of d -galacturonic acid (GalA, 81.0%), l -rhamnose (Rha, 2.6%), l -arabinose (Ara, 2.5%), and d -galactose (Gal, 3.6%) residues. Apiuman was found to be digestible with α-1,4- d -galacturonase to yield d -GalA, thus confirming that apiuman represents a pectic polysaccharide. The purified apiuman (AG) obtained was subjected to ion exchange chromatography on DEAE-cellulose to produce two polysaccharide fractions, AG-1 and AG-2, which had similar sugar compositions. Partial acid hydrolysis of apiuman, AG, revealed galacturonan to be at the core of the macromolecule. Nuclear magnetic resonance (NMR) spectra indicated that the backbone of apiuman appeared to represent α-1,4- d -galacturonan, with considerable amounts of l -Rha residues involved by α-1,2-linkages in the linear chain of the macromolecule core. The side chains were found to consist of Ara and Gal attached at the 4-position of the Rha residues. Furthermore, apiuman, AG, was found to improve the survival of mice subjected to a lethal dose of lipopolysaccharide (LPS), and the anti-endotoxemic effect of apiuman, AG, was shown to be mediated by decreased interleukin-1β (IL-1β and increased interleukin-10 (IL-10) production. Apiuman was also shown to diminish the amount of neutrophils migrating to the peritoneal cavity after LPS injection.

We studied the effects of bone marrow cell transplantation on myocardium of the prenecrotic zone in Wistar rats. Intramyocardial cell transplantation reduced the severity of hypertrophy of myocardium and the degree of its cicatricial degeneration on day 40 after cryodestruction. The morphology of the myocardium in the prenecrotic zone depended on the type of transplanted cells. The course of inflammation was swifter; vascularization of the myocardium was more intensive. The best effect, evaluated by the number of new vessels, was observed after MSC transplantation. Hence, the positive effect of bone marrow cell transplantation is realized at the expense of more rapid structural organization of the damaged site and stimulation of myocardial vascularization.

Deepwater sections of the Maikop Group (Oligocene-lower Miocene) and overlying lower-middle Miocene sediments are studied near the Perekishkyul Settlement in lower reaches of the Sumgait River, northern Azerbaijan. Several lithological reference levels and the Spiratella (Pteropoda) Beds are used to correlate preliminarily these uniform clayey sections barren of benthic fossils. Based on distribution of the organicwalled and calcareous phytoplankton, spores, pollen and ichthyofossils, the sections are first subdivided in detail and dated. Distinguished in the sections are two regional levels characterizing distortions in the basin hydrological regime during the late Oligocene (level with large Leiosphaeridia and Pterospermella) and Burdigalian (level with dominant dinocysts of the genus Batiacasphaera), and separate intervals with diverse phytoplankton and ichthyofossils. The Tarkhanian sediments are marked by first-appearing oceanic nannoplankton with Sphenolithus heteromorphus and deepwater ichthyofauna with Vinciquerria merklini. The Karaganian is established at the first occurrence level of peculiar endemics (Sardinella karaganica W. Dan. and Mugil karaganicus Swich.) among ichthyofauna.

We studied the presence of colony-forming cells in cell culture from rat heart 40 days after experimental myocardial infraction. The mean cellularity in this pathology was 12+/-8 cell/cm2, which is 20-fold lower than in intact myocardium. Transplantation of mesenchymal stem cells into the remodeling myocardium restored the pool of colony-forming cells. This effect depended on the state of transplanted cells. After transplantation of mesenchymal stem cells with low content of stress proteins, 6+/-2 colonies were detected, while after transplantation of cells with high content of hsp70 and hsp60 stress proteins (modified mesenchymal stem cells) 18+/-5 colonies were found, the mean cellularity of the corresponding cultured being 946+/-267 and 1926+/-123 cell/cm2. The positive effect of modified mesenchymal stem cells was observed on days 4 and 7 after transplantation. We conclude that postinfarction remodeling mobilized the total pool of regional stem cells; mesenchymal stem cells with high content of hsp70 and hsp60 demonstrated highest survival rate after intramyocardial transplantation.

We studied the effect of pectin gel particles on endotoxemia in mice induced by restraint stress. It was shown that the concentration of LPS in mouse blood increased during restraint stress, which was associated with memory impairment. Pectin gel particles prevented the development of stress-induced endotoxemia and memory impairment in mice.

I provide a model that connects the bribe amount with the fines imposed on both bribe-taker and bribe-payer. I show that Basu (2011) proposal to not punish bribe-payers to induce whistle-blowing does not have to help lower bribes. Higher fines on bribe-takers will make them ask for larger bribes, whereas lowering fines for bribe-paying on bribe-payers might increase their willingness to pay the bribe. An increase in transparency, if achieved, effectively increases both fines simultaneously.

We study how ownership structure and management objectives interact in determining the company size without assuming information constraints or any explicit costs of management. In symmetric agent economies, the optimal company size balances the returns to scale of the production function and the returns to collaboration efficiency. For a general class of payoff functions, we characterize the optimal company size, and we compare the optimal company size across different managerial objectives. We demonstrate the restrictiveness of common assumptions on effort aggregation (e.g., constant elasticity of effort substitution), and we show that common intuition (e.g., that corporate companies are more efficient and therefore will be larger than equal-share partnerships) might not hold in general.

Engraulis encrasicolus and Sardina pilchardus stocks are highly variable in terms of recruitment, biomass and spatial distribution. Changes in habitat conditions may influence both the survival of the early life stages and the adult stages. Detailed studies on the spatial distribution and habitat selection of such species have been performed in different areas of the world, highlighting the importance of environmental processes. The present study analyzes the spatial distribution of anchovy and sardine in the Tyrrhenian Sea in relation to environmental heterogeneity. Four acoustic surveys were carried out in this area in the period 2009–2014. Analysis of the environmental dataset permitted identification, in two specific areas, of a pattern of variables driving enrichment processes and impacting on the habitat suitability of the two species. In the northern and central parts of the study area, both anchovy and sardine showed a marked preference for shallower areas characterized by lower salinity. In these areas, PCA results on an environmental dataset highlighted a strong link between primary production, particulate organic carbon, distance from the mouth of the river, salinity and depth. A less clear picture was obtained for the southern part of the Tyrrhenian sea, characterized by a narrow continental shelf, moderately complex coastline morphology and the presence of very small rivers. Most of the anchovy biomass was found to be located in enclosed areas (gulfs) under the influence of relatively small rivers. This finding, taking into account that the surveys were carried out during the anchovy spawning period, highlights for such species a positive effect of the interaction between coastal morphology and riverine input, probably favoring food supply and retention of spawning products.

Cell morphogenesis requires dynamic communication between actin filaments and microtubules which is mediated, at least in part, by direct structural links between the two cytoskeletal systems. Here, we examined interaction between the CLIP-associated proteins (CLASP) CLASP1 and CLASP2, and actin filaments. We demonstrate that, in addition to a well-established association with the distal ends of microtubules, CLASP2a co-localizes with stress fibers, and that both CLASP1a and CLASP2a co-immunoprecipitate with actin. GFP-CLASP2a exhibits retrograde flow in the lamellipodia of Xenopus primary fibroblasts and in the filopodia of Xenopus spinal cord neurons. A deletion mapping analysis reveals that both the microtubule-binding domain of CLASP2 (which is homologous between all CLASPs) and the N-terminal dis1/TOG domain of CLASP2a (which is homologous between a isoforms) possess actin-binding activity. Fluorescence resonance energy transfer experiments demonstrate significant energy transfer between YFP-CLASP2a and CFP-actin. Our results indicate that CLASPs function as actin/microtubule crosslinkers in interphase cells. We propose that CLASPs facilitate recognition of actin filaments by the plus ends of growing microtubules at the initial stages of actin-microtubule interaction. Cell Motil. Cytoskeleton 64: 519‐530, 2007. ' 2007 Wiley-Liss, Inc.

The pectic polysaccharide named rauvolfian RS was obtained from the dried callus of Rauvolfia serpentina L. by extraction with 0.7% aqueous ammonium oxalate. Crude rauvolfian RS was purified using membrane ultrafiltration to yield the purified rauvolfian RSP in addition to glucan as admixture from the callus, with molecular weights 300 and 100–300 kD, respectively. A peroral pretreatment of mice with the crude and purified samples of rauvolfian (RS and RSP) was found to decrease colonic macroscopic scores, the total area of damage, and tissue myelope roxidase activity in colons as compared with a colitis group. RS and RSP were shown to stimulate production of mucus by colons of the colitis mice. RSP appeared to be an active constituent of the parent RS. The glucan failed to possess anti-inflammatory activity.

The pectic polysaccharide named potamogetonan (PN) was obtained using extraction of the leaves and stems of the common pondweed Potamogeton natans L. by an aqueous ammonium oxalate. The purified potamogetonan PN-300 was obtained using membrane ultrafiltration of PN and proved to be pectin with a molecular weight of 300 kDa. The capacity of potamogetonan PN-300 to prevent inflammation was assessed using a carrageenan paw edema test in mice. Oral administration of PN-300 24 h prior to induction of inflammation was found to reduce edema formation in a dose-related manner. The maximal effect of PN-300 was observed at 1 h after carrageenan injection (60% reduction of footpad swelling) and was comparable to that of indomethacin. The delayed edema (5 h) was less affected by pre-administration of PN-300 (33% reduction). PN-300 was found to improve the survival of mice subjected to a lethal dose of LPS. The anti-endotoxemic effect of PN-300 was shown to be mediated by decreased TNF-α and IL-1β and increased IL-10 production. Thus, a pectin named potamogetonan PN-300 was isolated from P. natans and was shown to possess a preventive antiinflammatory effect following oral administration. Copyright © 2007 John Wiley & Sons, Ltd.

Background The meningeal hemangiopericytoma (MHPC) is a vascular tumor arising from pericytes. Most intracranial MHPCs resemble meningiomas (MNGs) in their clinical presentation and histological features and may therefore be misdiagnosed, despite important differences in prognosis. Materials and methods We report 8 cases of MHPC and 5 cases of MNG collected from 2007 to 2011 from the Neuro-Surgery and Histopathology departments. All 13 samples were re reviewed by two independent pathologists and investigated by immunohistochemistry (IHC) using mesenchymal, epithelial and neuro-glial markers. Additionally, we screened all tumors for a large panel of chromosomal alterations using multiplex ligation probe amplification (MLPA). Presence of the NAB2-STAT6 fusion gene was inferred by immunohistochemical staining for STAT6. Results Compared with MNG, MHPCs showed strong VIM (100% of cases), CD99 (62%), bcl-2 (87%), and p16 (75%) staining but only focal positivity with EMA (33%) and NSE (37%). The p21 antibody was positive in 62% of MHPC and less than 1% in all MNGs. MLPA data did not distinguish HPC from MNG, with PTEN loss and ERBB2 gain found in both. By contrast, STAT6 nuclear staining was observed in 3 MHPC cases and was absent from MNG. Conclusions MNG and MHPC comprise a spectrum of tumors that cannot be easily differentiated based on histopathology. The presence of STAT6 nuclear positivity may however be a useful diagnostic marker.

The aims of the study were to evaluate the effects of renal sympathetic denervation (RSD) on the heart and to identify the predictors of RSD efficacy in patients with resistant arterial hypertension. The study comprised 60 RSD patients (54.6 ± 9.5 years) who received full-dose antihypertensive therapy (4.1 drugs) including diuretics. Initially, 58.6 % of patients had abnormal left ventricular (LV) diastolic function. All patients received echocardiography before and 24 weeks after RSD. Renal sympathetic denervation was achieved through the endovascular radiofrequency ablation (RFA) of the renal arteries. Drug therapy continued for the entire period of observation. After RSD, all patients were retrospectively assigned to two groups: group 1 comprised patients (n = 22; 36.7 %) in whom the myocardial mass (MM) of the left ventricle decreased by more than 10 g after RSD; group 2 comprised patients (n = 38; 63.3 %) in whom LV MM increased or decreased by less than 10 g. Anthropometry, arterial blood pressure, heart rate, therapy, and LV end-diastolic dimensions (EDD) were comparable in these groups. After RSD, the values of office blood pressure significantly decreased and MM regressed by more than 10 g in 36.7 % of patients; LV diastolic function normalized in 31 % of patients, and diastolic dysfunction improved in 14 % of patients. The study found the associations between the initial LV wall dimensions and LV MM changes. Unlike LV EDD, arterial blood pressure, or heart rate, the initial values of LV wall thickness predicted LV MM regress. Trial registration: #NCT01499810 https://clinicaltrials.gov/ct2/show/NCT01499810 .

A major challenge to improve outcomes in paediatric GBM and DIPG is their extensive intratumoral heterogeneity, reflected by differing cellular morphologies and genomic imbalances present within an individual sample. This is apparent also at the mutational level, where a complex subclonal architecture can be inferred from bulk tumour sequencing at diagnosis/biopsy and at relapse/autopsy. Spatial heterogeneity is also observed by studying multiple topographically distinct regions from the same tumour. We sought to understand the relative functional contributions of these subclonal cell populations to tumour development and evolution. We have established 20 primary patient-derived cultures (10 pGBM, 10 DIPG), with extensive phenotypic and genomic characterisation to ensure accurate recapitulation of the original disease. Single cells were flow-sorted into individual wells in a 96-well format and allowed to form colonies under stem cell conditions on laminin-coated plates (2D) or as neurospheres (3D). Individual colonies were subjected to high-throughput image analysis and targeted re-sequencing. Clonogenicity ranged from 0-20%, significantly higher in H3.3K27M mutant cells. Such an approach allowed for the isolation of single stem cell-derived colonies harbouring clonal mutations, some of which were present in only a small proportion of the original sample, such as a truncating mutation in the histone H4K20 methyltransferase SUV420H1 in a DIPG sample, originally found in 2/678 reads by targeted resequencing. Comparing colonies harbouring this mutation with their ‘natural isogenic’ counterparts with an otherwise identical genetic background, we observed significantly increased invasion and migration in the SUV420H1 mutant cells. Notably, bulk cultures containing mixtures of these distinct subpopulations were found to have additionally enhanced pro-tumorigenic potential than either colony alone. These data identify a distinct genetic and functional diversity in paediatric glioma stem cell populations, and suggest a co-operativity of tumour subclones. Treatment strategies aimed at disrupting these interactions may represent a novel therapeutic approach in these diseases.

Recent molecular profiling studies of paediatric high grade (pHGG) and diffuse intrinsic pontine glioma (DIPG) have refined these tumours into age- and location-based subgroups driven by unique genetic and epigenetic alterations, however individual studies are underpowered to investigate subgroup-specific events. We have retrieved publicly available genome-wide data from ∼560 pHGG/DIPG samples and combined this with ∼140 unpublished cases including young adults up to the age of 30 years. We have integrated multiple array-based and sequencing platforms to produce DNA copy number profiles from ∼700 tumours, ∼500 with clinicopathological and histone H3 annotation, and >300 of which have full somatic sequence information. We identified subgroup-specific genetic alterations co-segregating, or mutually exclusive, with known driving histone mutations. H3.3G34R/V cerebral hemispheric tumours harbour significantly more CNAs and SNVs than other subgroups. These included novel amplified loci at 1p13.3 (KCNA) and a histone cluster at 6p22.2, though lacked key amplified loci such as MYC/MYCN. H3.1K27M DIPG were distinct from H3.3K27M primarily on the basis of whole arm chromosomal changes (enriched +2, -16q; reduced -17p and a lack of TP53 mutations). H3.3K27M tumours harboured specifically enriched known (7q31.2;MET) and novel amplicons (17p11.2;TOP3A). Integration with mutation data identified subgroup-independent, non-overlapping pathway-level recurrent alterations, such as RTK-PI3K-mTOR, dysregulated in 55% cases and conferring shorter survival in hemispheric, but not other locations. As well as inter-tumoral differences, deeper interrogation of the sequencing data also reveals substantial intra-tumoral heterogeneity. Whilst driving histone H3 mutations were found to be present in 100% of cells, mutations in genes such as PDGFRA and PIK3CA were predominantly found at subclonal levels, and tumours from all locations were inferred to be comprised of multiple subclonal populations, with H3.3G34R/V the most genetically diverse. These data improve our understanding of the underlying biology of pHGG/DIPG, and will provide rational targets for subgroup-specific and –independent therapies.

Approximately 1% of all paediatric brain tumours occur at less than one year of age, and glioblastoma (GBM) account for only 2-9% of these. Their clinical presentation, location, histology, behaviour, response to therapy and outcome differ from GBMs of older children and adults, and they do not appear to be associated with known histone H3 or IDH1-driven subgroups. Recent genome and RNA sequencing data highlighted recurrent NTRK1-3 fusions in 4/10 non-brainstem GBM under the age of 3 years, including one case of ETV6-NTRK3 in a cerebral hemispheric tumour diagnosed at 1 month old. These fusions are usually associated with congenital mesoblastic nephroma and congenital infantile fibrosarcoma, and upon review of a series of cases diagnosed as glioblastoma under the age of 1yr, 6/14 were found to present with a predominant spindle cell morphology, a relatively circumscribed interface with adjacent brain, and without prominent infiltration. These tumours were positive for GFAP as well as desmin, CD56 and CD99. One case was diagnosed in utero by ultrasound, and at birth underwent surgical resection from which primary cell cultures were established under stem cell conditions on laminin-coated plates. Exome and RNA sequencing revealed the presence of the ETV6:NTRK3 fusion transcript as the sole somatic alteration, validated by RT-PCR, Sanger sequencing and FISH. 450K methylation profiling clustered the tumour with desmoplastic infantile ganglioglioma rather than subgroups of paediatric GBM. Primary cells treated with the CNS-penetrant small molecule Trk inhibitor PHA-848125, resulted in marked sensitivity (GI50 = 200nM) compared to cerebral hemispheric (H3 wild-type) and thalamic (H3.3K27M) paediatric GBM cells (GI50 = 1.8-4.6mM). A subset of infantile/congenital brain tumours classified as GBM appear to represent a distinct entity more closely related, biologically and histologically, to mesenchymal cancers. Agents targeting ETV:NTRK3 may be a useful novel treatment strategy in these children.

Here we review available data on nitric oxide (NO)-mediated signaling in skeletal muscle during physical exercise. Nitric oxide modulates skeletal myocyte function, hormone regulation, and local microcirculation. Nitric oxide underlies the therapeutic effects of physical activity whereas the pharmacological modulators of NO-mediated signaling are the promising therapeutic agents in different diseases. Nitric oxide production increases in skeletal muscle in response to physical activity. This molecule can alter energy supply in skeletal muscle through hormonal modulation. Mitochondria in skeletal muscle tissue are highly abundant and play a pivotal role in metabolism. Considering NO a plausible regulator of mitochondrial biogenesis that directly affects cellular respiration, we discuss the mechanisms of NO-induced mitochondrial biogenesis in the skeletal muscle cells. We also review available data on myokines, the molecules that are expressed and released by the muscle fibers and exert autocrine, paracrine and/or endocrine effects. The article suggests the presence of putative interplay between NO-mediated signaling and myokines in skeletal muscle. Data demonstrate an important role of NO in various diseases and suggest that physical training may improve health of patients with diabetes, chronic heart failure, and even degenerative muscle diseases. We conclude that NO-associated signaling represents a promising target for the treatment of various diseases and for the achievement of better athletic performance.

The aim of the study was to evaluate the impact of idiopathic ventricular tachycardia and premature ventricular beats on cardiac function and dyssynchrony and to elucidate relationships between data of scintigraphic and intracardiac electrophysiology studies (EPSs).The study comprised 64 patients with idiopathic ventricular arrhythmias (VAs; median age of 14 years ranging from 8 to 18 years). The control group comprised 20 patients (median age of 12 ranging from 7 to 16 years) without cardiac arrhythmias. EPS and radiofrequency ablation (RFA) procedure for VA were performed in 21 children according to indications. The functional state of both ventricles was assessed by gated blood pool single photon emission computer tomography (GBP-SPECT) before and after RFA in all patients.Patients with VA had local areas of asynchronous myocardial contraction (AMC). Compared with the control group, VA patients had significantly higher values of end-diastolic volume, end-systolic volume, and lower contractility indices. Negative association was found between total numbers of AMC areas and cardiac contractility indices. Ectopic foci localization, determined based on EPS data, was associated with AMC areas topography based on GBP-SPECT. RFA procedure significantly improved cardiac contractility indices; AMC areas completely disappeared or decreased compared with the preoperative conditions.In VA patients, AMC areas were localized mostly in the right ventricle. Comparison of the results of GBP-SPECT and EPS studies showed a relationship between AMC localizations and ectopic foci topography. The fact that AMC areas disappeared after RFA supports the hypothesis stating that the presence of AMC areas is a scintigraphic symptom of ectopic focus.

Tenure-track contract is criticized for curbing the incentives for spending effort after obtaining the tenured status. Yet, the most productive faculty seems to work on a tenure contract, and schools who aim to employ the best faculty seem to prefer to offer tenure-track contracts to their new hires. I argue that tenure-track contracts are by construction more attractive to more able freshly minted PhDs, and therefore the observed sorting is rationalizable.

The aim of the paper is to present the results of a study on the relationship between ichthyoplanktonic distribution and the hydrographic features in the Cape Passero area (Strait of Sicily). Acoustic, physical, and biological data were collected during two multidisciplinary research surveys performed during the summer in 2002 and 2003 in the Strait of Sicily. The oceanographic surveys ‘Ansic 02’ and ‘Ansic 03’ were carried out on board the RV Urania in the framework of the work programme of the ASTAMAR research project. An accurate post-processing procedure was adopted to estimate anchovy larvae distribution in a sea area around Cape Passero, also taking into account the results of biological sampling. The analysis of acquired data has singled out a relationship between the acoustic-based ichthyoplanktonic distribution and the presence in both surveys of a thermohaline front positioned in the south-eastern part of the study area, which appears to be able to promote concentration processes.

Lemnan LM, apiogalacturonanic pectin of duckweed Lemna minor L. was tested for adjuvant properties following oral administration with protein antigen. Male Swiss mice were orally immunized thrice with weekly intervals with free OVA or OVA with lemnan (LM). Lemnan LM was shown to increase delayed type hypersensitivity (DTH) and serum anti OVA IgG responses. LM was established to increase levels of both serum IgG1 and IgG2a subclasses, intestinal IgA and failed to elevate levels of serum IgE. Lemnan was found to increase the adhesion of macrophages and to enhance the generation of oxygen radicals by macrophages in response to phorbol 12-myristate 13-acetate. Serum OVA levels were four-fold higher in mice immunized with the mixture of OVA and LM in comparison with those in mice immunized with OVA only. Thus, substantial systemic and local mucosal immune responses were attained by oral immunization with the mixture of OVA and lemnan. Lemnan appeared to elicit adjuvant activity via induction of both Th1- and Th2-type responses. The immunopotentiating effect of lemnan may result from enhanced antigen ingestion and stimulation of macrophage activity.

Galacturonan, the main constituent of the backbone (core) of the comaruman macromolecule, a pectin from the marsh cinquefoil Comarum palustre L., was obtained on partial acid hydrolysis of the pectin. Using atomic force microscopy and methylation analysis of the galacturonan, the backbone of the comaruman macromolecule was shown to contain branches as side chains consisting of alpha-1,4-linked residues of D-galactopyranosyl uronic acid attached to the 2- and 3-positions of the galacturonic acid residues of the core, in addition to linear regions of alpha-1,4-D-galacturonan. A few side chains appear to attach to 2,3-positions of the D-galacturonic acid residues.

An effect of apiogalacturonanic pectin of duckweed Lemna minor L. (lemnan LM) was studied on the inflammatory response to ovalbumin injected intradermally into the footpad of control and ovalbumin-fed mice. Lemnan LM (1–2 mg per mouse) was found to enhance by as much as 50–60% the footpad swelling in control mice. Oral administration of ovalbumin was shown to result in sensitization that increased inflammation. Ovalbumin admixed with lemnan was found to increase by two-fold footpad edema in comparison with the mice receiving ovalbumin alone. Apple pectin used as a reference compound failed to influence the inflammatory reaction. Degradation of lemnan was performed to elucidate the active region of the polysaccharide macromolecule. The apiogalacturonanic fragment (LMP) obtained using a digestion of lemnan LM with pectinase was shown to increase the footpad response in both control and ovalbumin-fed mice. Fragment LMPH deprived of some terminal apiose residues as a result of partial acidic hydrolysis failed to have an effect on the inflammatory response. Thus, the data obtained reveal an enhancement by lemnan of the inflammatory response. The ramified apiogalacturonan seemed to be the active region of the lemnan macromolecule. Copyright © 2006 John Wiley & Sons, Ltd.

The effects of Amiodarone and Nibentan on the postextrasystolic and postrest reactions were stud- ied with normal rat myocardium and (for Amiodarone only) with myocardial strips isolated from patients with coronary heart disease (CHD). One-third of the patient samples displayed spontaneous activity that could be effectively suppressed by Amiodarone. All other samples displayed postrest inotropic effects of two types, with 30 or 70% decreases in the parameters of the contraction-relaxation cycle. Amiodarone did not influence the reaction of the first type but increased the postrest inotropic response of the second type. Both Amiodarone and Nibentan decreased the postextrasystolic inotropic response of the normal rat myocardium and simultaneously increased the postextrasystolic contraction. In addition, these drugs nearly doubled postrest potentiation. In the patient myocardium samples, however, Amiodarone did not decrease the amplitude of the extrasystolic contrac- tion but increased postextrasystolic potentiation. It was supposed that, in the myocardium of some CHD patients (as well as in the rat myocardium), Amiodarone increases the ability of the sarcoplasmic reticulum to accumulate calcium ions. The same was supposed for the effects observed for Nibentan in tests with the rat myocardium. This phenomenon may contribute to the suppression of abnormal activity of the myocardium and strengthen the direct antiarrhythmic effect of Amiodarone.

Earlier, we detected antiinflammatory action of comaruman, pectin of the marsh cinquefoil Comarum palustre L. This effect can be explained by new data concerning inhibition of adhesion of human neutrophils to fibronectin by comaruman and its fragments. The galacturonan backbone fragment of molecular mass >10 kD appears to be the active region of the comaruman macromolecule. Comaruman CP (50–200 μg/ml) was found to decrease adhesion of neutrophils stimulated by phorbol 12-myristate 13-acetate (PMA, 1.625 μM) and by dithiothreitol (DTT, 0.5 mM). The fragments of comaruman CP-H (100 kD), CP-H1 (10–50 kD), and CP-H2 (100 kD) obtained by acidic hydrolysis and representing regions of linear polygalacturonan are shown to inhibit neutrophil adhesion more than the crude pectin. A fragment CP-E (

The pectic polysaccharide named bergenan BC was obtained using extraction of the green leaves of Siberian tea Bergenia crassifolia (L.) Fritsch. by an aqueous ammonium oxalate. The polysaccharide obtained was proved to comprise mainly d-galacturonic acid, galactose, rhamnose, arabinose and glucose residues and appeared to be pectin. Delayed type hypersensitivity (DTH) reaction to aggregated ovalbumin (agOVA) was found to increase in mice that received bergenan solution (2 mg/mL) for 3 weeks. Bergenan BC was observed to enhance the uptake capacity of human neutrophils at a concentration 100 microg/mL and to stimulate the generation of oxygen radicals by mouse peritoneal macrophages in vitro. Bergenan BC was found to increase the spontaneous adhesion of peritoneal leukocytes and failed to influence adhesion stimulated by PMA or adhesion of peritoneal leukocytes incubated in the presence of 5 mm EDTA. Bergenan failed to show cytotoxic action. The viability of peritoneal leukocytes was estimated to be equal to 91% +/- 8% and 90% +/- 7%% in the control and in the pectin solution at a concentration of 1 mg/mL.Thus, bergenan was shown to possess immunostimulating activity in relation to DTH response in vivo and phagocytic activity in vitro.

Cardiomyocyte energy metabolism in experimental unfolding postinfarction cardiosclerosis and diabetes mellitus was studied. Postinfarction cardiosclerosis formed 6 weeks after coronary artery occlusion. Diabetes mellitus was induced by intraperitoneal injection of streptozotocin (60 mg/kg). The rate of oxygen consumption in postinfarction cardiosclerosis and diabetes increased by 3.4 and 4.2 times, respectively. Stimulation of mitochondrial respiration (ATP, palmitic acid) significantly increased oxygen consumption in animals with postinfarction cardiosclerosis and significantly reduced this process in diabetes. The content of LDH and SDH in the myocardium of animals with diabetes and postinfarction cardiosclerosis was significantly below the control. Hence, the development of postinfarction cardiosclerosis and diabetes mellitus were characterized by reduced generation of ATP in anaerobic and aerobic pathways and oxidative phosphorylation in cardiomyocytes.

Tau is a major microtubule-associated protein which induces bundling and stabilization of axonal microtubules (MTs). To investigate the interaction of tau with MTs in living cells, we expressed GFP-tau fusion protein in cultured Xenopus embryo neurons and performed time-lapse imaging of tau-labeled MTs. Tau uniformly labeled individual MTs regardless of their assembly/disassembly status and location along the axon. Photobleaching experiments indicated that interaction of tau with MTs is very dynamic, with a half-time of fluorescence recovery of the order of 3 seconds. Treatment of cells with taxol, a drug that suppresses MT dynamics, rapidly induced detachment of tau from MTs. Although binding of tau to straight MTs was uniform, there was a heightened concentration of tau at the sites of high MT curvature. Our results suggest that dynamic interaction of tau with MTs may modify local mechanical properties of individual MTs and play a crucial role in the remodeling of the MT cytoskeleton during neuronal plasticity.

Changes in intracellular Ca2+ homeostasis induced by 1 μM Amiodarone solution were studied with muscle strips isolated from the right atrial auricles of patients with ischemic heart disease (IHD). The intracellular Ca2+ homeostasis was evaluated through the changes in contractility in the muscle strips caused by a 60-s discontinuation of their stimulation by electrical pulses. It was found that, in 30% of the IHD patients, the atrial myocardium displayed an abnormal contractility. Myocardium samples with normal contractility displayed two types of inotropic response to the resumption of electrical stimulation. The first type was associated with a 30% decrease in the parameters of the contraction–relaxation cycle, whereas, for the second type, this decrease was stronger (by 70% or more). An incubation of the muscle samples with the Amiodarone solution suppressed the abnormal contractility, did not affect the muscles with the first type of response, and significantly (p < 0.05) increased the inotropic response in the muscles with the second type of reaction to a short interruption in their electrical stimulation. As Amiodarone itself induces no direct positive inotropic effects, it was supposed that, in some cases of IHD, the presence of Amiodarone in the atrial myocardium is associated with an increased ability of the sarcoplasmic reticulum to retain calcium ions accumulated. This phenomenon may contribute to the suppression of abnormal activity of the atrial myocardium and strengthen the direct antiarrhythmic effect of Amiodarone.