YAP and TAZ in Vascular Smooth Muscle Confer Protection Against Hypertensive Vasculopathy.

BACKGROUND: Hypertension remains a major risk factor for cardiovascular diseases, but the underlying mechanisms are not well understood. We hypothesize that appropriate mechanotransduction and contractile function in vascular smooth muscle cells are crucial to maintain vascular wall integrity. The Hippo pathway effectors YAP (yes-associated protein 1) and TAZ (WW domain containing transcription regulator 1) have been identified as mechanosensitive transcriptional coactivators. However, their role in vascular smooth muscle cell mechanotransduction has not been investigated in vivo. METHODS: We performed physiological and molecular analyses utilizing an inducible smooth muscle-specific YAP/TAZ knockout mouse model. RESULTS: Arteries lacking YAP/TAZ have reduced agonist-mediated contraction, decreased myogenic response, and attenuated stretch-induced transcriptional regulation of smooth muscle markers. Moreover, in established hypertension, YAP/TAZ knockout results in severe vascular lesions in small mesenteric arteries characterized by neointimal hyperplasia, elastin degradation, and adventitial thickening. CONCLUSIONS: This study demonstrates a protective role of YAP/TAZ against hypertensive vasculopathy.

Loss of Vascular Myogenic Tone in miR-143/145 Knockout Mice Is Associated With Hypertension-Induced Vascular Lesions in Small Mesenteric Arteries.

OBJECTIVE: Pressure-induced myogenic tone is involved in autoregulation of local blood flow and confers protection against excessive pressure levels in small arteries and capillaries. Myogenic tone is dependent on smooth muscle microRNAs (miRNAs), but the identity of these miRNAs is unclear. Furthermore, the consequences of altered myogenic tone for hypertension-induced damage to small arteries are not well understood. APPROACH AND RESULTS: The importance of smooth muscle-enriched microRNAs, miR-143/145, for myogenic tone was evaluated in miR-143/145 knockout mice. Furthermore, hypertension-induced vascular injury was evaluated in mesenteric arteries in vivo after angiotensin II infusion. Myogenic tone was abolished in miR-143/145 knockout mesenteric arteries, whereas contraction in response to calyculin A and potassium chloride was reduced by ≈30%. Furthermore, myogenic responsiveness was potentiated by angiotensin II in wild-type but not in knockout mice. Angiotensin II administration in vivo elevated systemic blood pressure in both genotypes. Hypertensive knockout mice developed severe vascular lesions characterized by vascular inflammation, adventitial fibrosis, and neointimal hyperplasia in small mesenteric arteries. This was associated with depolymerization of actin filaments and fragmentation of the elastic laminae at the sites of vascular lesions. CONCLUSIONS: This study demonstrates that miR-143/145 expression is essential for myogenic responsiveness. During hypertension, loss of myogenic tone results in potentially damaging levels of mechanical stress and detrimental effects on small arteries. The results presented herein provide novel insights into the pathogenesis of vascular disease and emphasize the importance of controlling mechanical factors to maintain structural integrity of the vascular wall.

Regulation of microRNA expression in vascular smooth muscle by MRTF-A and actin polymerization.

The dynamic properties of the actin cytoskeleton in smooth muscle cells play an important role in a number of cardiovascular disease states. The state of actin does not only mediate mechanical stability and contractile function but can also regulate gene expression via myocardin related transcription factors (MRTFs). These transcriptional co-activators regulate genes encoding contractile and cytoskeletal proteins in smooth muscle. Regulation of small non-coding microRNAs (miRNAs) by actin polymerization may mediate some of these effects. MiRNAs are short non-coding RNAs that modulate gene expression by post-transcriptional regulation of target messenger RNA. In this study we aimed to determine a profile of miRNAs that were 1) regulated by actin/MRTF-A, 2) associated with the contractile smooth muscle phenotype and 3) enriched in muscle cells. This analysis was performed using cardiovascular disease-focused miRNA arrays in both mouse and human cells. The potential clinical importance of actin polymerization in aortic aneurysm was evaluated using biopsies from mildly dilated human thoracic aorta in patients with stenotic tricuspid or bicuspid aortic valve. By integrating information from multiple qPCR based miRNA arrays we identified a group of five miRNAs (miR-1, miR-22, miR-143, miR-145 and miR-378a) that were sensitive to actin polymerization and MRTF-A overexpression in both mouse and human vascular smooth muscle. With the exception of miR-22, these miRNAs were also relatively enriched in striated and/or smooth muscle containing tissues. Actin polymerization was found to be dramatically reduced in the aorta from patients with mild aortic dilations. This was associated with a decrease in actin/MRTF-regulated miRNAs. In conclusion, the transcriptional co-activator MRTF-A and actin polymerization regulated a subset of miRNAs in vascular smooth muscle. Identification of novel miRNAs regulated by actin/MRTF-A may provide further insight into the mechanisms underlying vascular disease states, such as aortic aneurysm, as well as novel ideas regarding therapeutic strategies. This article is part of a Special Issue entitled: ECS Meeting edited by Claus Heizmann, Joachim Krebs and Jacques Haiech.

Patients with bicuspid and tricuspid aortic valve exhibit distinct regional microrna signatures in mildly dilated ascending aorta.

MicroRNAs are able to modulate gene expression in a range of diseases. We focused on microRNAs as potential contributors to the pathogenesis of ascending aorta (AA) dilatation in patients with stenotic tricuspid (TAV) or bicuspid aortic valve (BAV). Aortic specimens were collected from the 'concavity' and the 'convexity' of mildly dilated AAs and of normal AAs from heart transplant donors. Aortic RNA was analyzed through PCR arrays, profiling the expression of 84 microRNAs involved in cardiovascular disease. An in silico analysis identified the potential microRNA-mRNA interactions and the enriched KEGG pathways potentially affected by microRNA changes in dilated AAs. Distinct signatures of differentially expressed microRNAs are evident in TAV and BAV patients vs. donors, as well as differences between aortic concavity and convexity in patients only. MicroRNA changes suggest a switch of SMC phenotype, with particular reference to TAV concavity. MicroRNA changes potentially affecting mechanotransduction pathways exhibit a higher prevalence in BAV convexity and in TAV concavity, with particular reference to TGF-ß1, Hippo, and PI3K/Akt/FoxO pathways. Actin cytoskeleton emerges as potentially affected by microRNA changes in BAV convexity only. MicroRNAs could play distinct roles in BAV and TAV aortopathy, with possible implications in diagnosis and therapy.

Emerging roles of the myocardin family of proteins in lipid and glucose metabolism.

Members of the myocardin family bind to the transcription factor serum response factor (SRF) and act as coactivators controlling genes of relevance for myogenic differentiation and motile function. Binding of SRF to DNA is mediated by genetic elements called CArG boxes, found often but not exclusively in muscle and growth controlling genes. Studies aimed at defining the full spectrum of these CArG elements in the genome (i.e. the CArGome) have in recent years, unveiled unexpected roles of the myocardin family proteins in lipid and glucose homeostasis. This coactivator family includes the protein myocardin (MYOCD), the myocardin-related transcription factors A and B (MRTF-A/MKL1 and MRTF-B/MKL2) and MASTR (MAMSTR). Here we discuss growing evidence that SRF-driven transcription is controlled by extracellular glucose through activation of the Rho-kinase pathway and actin polymerization. We also describe data showing that adipogenesis is influenced by MLK activity through actions upstream of peroxisome proliferator-activated receptor γ with consequences for whole body fat mass and insulin sensitivity. The recently demonstrated involvement of myocardin coactivators in the biogenesis of caveolae, Ω-shaped membrane invaginations of importance for lipid and glucose metabolism, is finally discussed. These novel roles of myocardin proteins may open the way for new unexplored strategies to combat metabolic diseases such as diabetes, which, at the current incidence, is expected to reach 333 million people worldwide by 2025. This review highlights newly discovered roles of myocardin-related transcription factors in lipid and glucose metabolism as well as novel insights into their well-established role as mediators of stretch-dependent effects in smooth muscle. As co-factors for serum response factor (SRF), MKLs regulates transcription of genes involved in the contractile function of smooth muscle cells. In addition to mechanical stimuli, this regulation has now been found to be promoted by extracellular glucose levels in smooth muscle. Recent reports also suggest that MKLs can regulate a subset of genes involved in the formation of lipid-rich invaginations in the cell membrane called caveolae. Finally, a potential role of MKLs in non-muscle cells has been discovered as they negatively influence adipocyte differentiation.

Inhibition of Polyamine Uptake Potentiates the Anti-Proliferative Effect of Polyamine Synthesis Inhibition and Preserves the Contractile Phenotype of Vascular Smooth Muscle Cells.

Increased vascular smooth muscle cell (VSMC) proliferation is a factor in atherosclerosis and injury-induced arterial (re) stenosis. Inhibition of polyamine synthesis by α-difluoro-methylornithine (DFMO), an irreversible inhibitor of ornithine decarboxylase, attenuates VSMC proliferation with high sensitivity and specificity. However, cells can escape polyamine synthesis blockade by importing polyamines from the environment. To address this issue, polyamine transport inhibitors (PTIs) have been developed. We investigated the effects of the novel trimer44NMe (PTI-1) alone and in combination with DFMO on VSMC polyamine uptake, proliferation and phenotype regulation. PTI-1 efficiently inhibited polyamine uptake in primary mouse aortic and human coronary VSMCs in the absence as well as in the presence of DFMO. Interestingly, culture with DFMO for 2 days substantially (>95%) reduced putrescine (Put) and spermidine (Spd) contents without any effect on proliferation. Culture with PTI-1 alone had no effect on either polyamine levels or proliferation rate, but the combination of both treatments reduced Put and Spd levels below the detection limit and inhibited proliferation. Treatment with DFMO for a longer time period (4 days) reduced Put and Spd below their detection limits and reduced proliferation, showing that only a small pool of polyamines is needed to sustain VSMC proliferation. Inhibited proliferation by polyamine depletion was associated with maintained expression of contractile smooth marker genes. In cultured intact mouse aorta, PTI-1 potentiated the DFMO-induced inhibition of cell proliferation. The combination of endogenous polyamine synthesis inhibition with uptake blockade is thus a viable approach for targeting unwanted vascular cell proliferation in vivo, including vascular restenosis.

Post stroke depression.

INTRODUCTION: Comorbidity of depression and stroke significantly reduces the quality of life of patients after the stroke. Squeal after stroke also determines the quality of life and have impact on the occurrence of depression after the stroke. In our study we investigated the occurrence of depression in patients after different types and subtypes of stroke measured by the Hamilton scale compared to the level of disability measured by NIHSS scale. GOAL: The goal was to make a comparative analysis of depression after stroke, according to gender and age, side of the lesion and the severity of neurological deficit. MATERIAL AND METHODS: Material for our work are 210 patients with stroke treated at the Neurology Clinic, Clinical Center of Sarajevo University in 2012, 105 male and 105 female. The mean age of the patients was 67.12 +/- 9.5 years. Ischemic stroke was present in 65% cases. There was no statistically significant difference between ischemic and hemorrhagic stroke among genders. In case of hemorrhagic M-56.7%, F-43.3%; ischemic M-48.3%, F-51.7% (chi-square = 6.563, p = 0.082). Depression was more prevalent among younger patients (52-60 years) with 39.2% then in the group of older patients (61-70 years) with 32% of depressed. In relation to gender there was significantly more patients with depression among women compared to men (63.8:27.2%, chi-square = 14.38, p = 0.00019). Depression was more frequent in patients with stroke in the left hemisphere medial localization (63%). NIHSS scale average was 16.07 with the minimum of 11 and maximum of 22, F = 52.56, p = 0.001. CONCLUSIONS: We can conclude that depression after stroke is more frequent in younger patients, female patients, patients with localized stroke in the medial left hemisphere and with higher disability score.

Cutaneous silent period in the evaluation of small nerve fibres.

INTRODUCTION: High intensity cutaneous stimulus transiently suppresses tonic voluntary muscle activity resulting in cutaneous silent period (CSP). AIM: The aim of our study was to evaluate the normal values of an onset latency L1, a late latency L2 and a duration of CSP after stimulating sensory fibres of the median nerve. MATERIAL AND METHODS: This prospective study was performed at the Neurology Department, Clinical Center of Sarajevo University in period from January 1st 2013 to December 1st 2013. In our study we examined 61 subjects. The group included our relatives, coworkers and friends. The informed consent from testing subjects was obtained. RESULTS: The origin of silent period is stimulation of small A-delta nerve fibres. The pre-synaptic or post-synaptic interruption of the electrical volley to motor neurons is discussed. Median values of muscle activity suppression in healthy female is 55.0 ms (45.0-74.0) and 59.0 ms (52.0-67) male subjects. There is a correlation between the onset latency L1 and the late L2 latency (p < 0.03). In the on-going study it seems that delay of L1 and shorter muscle activity suppression might provide a sign of small nerve fibres involvement. CONCLUSION: The use of CSP improves the value of neurophysiology examination.

Suicidal ideation and thoughts of death in epilepsy patients.

BACKGROUND: Suicidal behavior is an important worldwide health problem. Psychiatric disorders, especially mood disorders, are the main risk factors for suicidal behavior. Suicide is an important cause of death in patients with epilepsy. The aim of this study was to analyze the presence of suicidal ideation in patients with epilepsy. SUBJECTS AND METHODS: The study included 50 epilepsy inpatients and outpatients of both genders, aged 18 years and older, treated at the Department of Neurology, Clinical Center University of Sarajevo in the period from 1(st) of April - October 1(st) 2007. The sample was selected randomly. Applied research instruments were general questionnaire, HAM-D-17, BHS and BSS. RESULTS: Suicidal ideation and thoughts of death were present in 38% epilepsy patients. Symptoms of depression as well as feelings of hopelessness were found in half of the participants (52% and 48%), and were significantly more common in epilepsy patients with suicidal ideation. There was a significant relation of suicidal ideation with the presence of chronic pain (3.86; p=0.49), sexual/physical abuse history (5.95, p=0.015), level of hopelessness (20.7; p=0.000) and severity of depression (14.48; p=0.000) in epilepsy patients. Multiple logistic regression analysis showed that unemployment (Exp(B) 33.9; p=0.007) and the level of hopelessness (Exp(B) 14.9; p=0.001) were independently related to suicidal ideation in these patients. CONCLUSIONS: The study has shown that the level of hopelessness and unemployment have a predictive value for appearance of suicidal ideation in epilepsy patients. In the prediction of suicidal ideation in this population of patients, there is no single variable that should be considered as specific and separate.

Valsartan in the treatment of hypertension.

INTRODUCTION: Hypertension represents an important public health problem. Effective treatment of hypertension is imperative for primary care. GOAL: The goal of this study was to examine the efficacy of Valsartan in the treatment of hypertension with emphasis on the overall efficacy in reduction of systolic and diastolic blood pressure in a sample of 738 patients. MATERIAL AND METHODS: The study lasted for 12 months (from January 1, 2012 until December 31, 2012 year) and conducted in 18 public health institutions in B&H. Parallel follow up of Valsartan antihypertensive effect through repeated measurements every three months was conducted. RESULTS AND DISCUSSION: Our results indicate that both systolic and diastolic blood pressure decreased significantly after 12 weeks of Valsartan treatment. Analysis of adverse effects did not showed statistical significance of side effects for total sample. Statistical analysis by Yates chi-square did not show the presence of statistically significant differences in adverse effects by gender. CONCLUSION: We conclude that Val or Val plus are effective and safe antihypertensive drugs for the treatment of mild to moderate

Predilection role diabetes mellitus and dyslipidemia in the onset of ischemic stroke.

INTRODUCTION: Stroke is the third leading cause of mortality, disability and dementia, but leading cause of epileptic manifestations in the elderly. Diabetes mellitus as permanently elevated blood glucose, often accompanied by dyslipidemia, is among the leading causes of atherosclerotic alteration in blood vessels and is also increasing in the world. GOAL: To determine the existence and predilection of diabetes mellitus and dyslipidemia, in the development of ischemic stroke. MATERIAL AND METHODS: During the 2011 are analyzed all people with stroke admitted at the Neurology Clinic. All patients underwent neurological tests and the laboratory test with special emphasis on the value of blood glucose and lipid levels, with brain CT which confirmed the existence of a stroke, EEG and internist examination. RESULTS: During the one-year period the stroke was confirmed in 1184 patients, aged 33-81 years and 37% in the younger age group (up to 50 yrs.). There was 50.67% male and 49.33% female patients. Ischemic stroke was confirmed in 78.0% (56% with thrombotic and 22% with embolic genesis), of which the 32% was lacunar infarcts (up to 1.5 cm) and hemorrhagic in 22% (SAH in 4.8%, and intracerebral hemorrhage in 17.2%). The most frequent risk factors were hypertension 85%, then smoking in 65%, diabetes mellitus in 39.0%, in 27.38% dyslipidemia, previous stroke in 26.69%, in 23.57% arrhythmia In the baseline sample 30.06% of patients had previously diabetes mellitus and in 8.94% the diabetes was diagnosed during hospitalization, while dyslipidemia was known from earlier in 22.0% and in 5.38% cases was detected during the hospitalization. Among treated patients 79.01% survived, while 20.09% have a fatal outcome. CONCLUSIONS: Diabetes mellitus and dyslipidemia, along with hypertension and smoking are the leading risk factors for the occurrence of stroke. By timely detection and treatment can be controlled slow atherosclerotic changes in blood vessels and thus prevent stroke.

Betaferon efficiency indicators in the treatment of relapsing remitting form of multiple sclerosis.

INTRODUCTION: Multiple sclerosis (MS) is the most common, chronic debilitating disease in young adults characterized by a wide variety of neurological symptoms and unpredictable increasing disability. Worldwide, MS affects about 2.5 million people, with a female-to-male ratio of approximately 2:1. The therapies used in the chronic treatment of MS are immune-modulating agents. Interferon beta -1b has been shown to decrease the rate of relapses, the burden of lesions seen on MRI, and the rate of accumulated disability. AIM: Determine the efficacy of Betaferon in patients with RR form of MS in terms of the degree of disability and the number of relapses during the two years of continuous treatment. SUBJECTS AND METHODS: The study, partly retrospective, partly prospective, included 58 patients of both sexes with MS, RR type, from the Federation of Bosnia and Herzegovina, who received Betaferon treatment, from the Solidarity Fund, during 2 years period. Evaluation of efficacy was based on the degree of disability measured by EDDS scale and number of relapses. RESULTS: In our sample, women were represented in the ratio of 3:1 compared to men. 44.8% of patients were referred from Clinical Centre University in Sarajevo (UCCS), 34.5% from University Clinical Centre Tuzla (UCCT) and 20.7% from Clinical Hospital Center Mostar (CHCM). The smallest number of patients have had a relapse sent from CCUS (0.04, SD = 0.196), which was in direct correlation with input EDSS score at baseline (= 1.3) compared to patients from the UCCT, who had an average of 1.05 relapses, SD = 1.35, and the input EDSS score is 2.15. Patients referred from CHCM had an average of 0.08 relapses, SD = 0.93, while the input EDSS score was around 1. In terms of the degree of disability, measured by EDSS, we get a minimal increase in the patients from UCCS and UCCT, while patients from CHCM had a reduction of EDDS for the 0.45 (p < 0.05). CONCLUSION: Betaferon therapy must start as soon as possible, preferably when clinically isolated syndrome (CIS) is diagnosed. The reason for early start is to delay the transfer of disease in to definite multiple sclerosis, and thereby reduce disability, which disease brings to young people. Key words:

Role of early viral infections in development of multiple sclerosis.

INTRODUCTION: Multiple sclerosis is a chronic inflammatory, autoimmune, demyelinating, disease but also degeneration of axons, with mainly progressive course, causing greater or lesser degree of disability. In addition to genetic predisposition the environmental factors, with particular importance of early viral infection, have an essential role in the development of MS. These are called long-acting viruses that remain hidden in the body for years by encouraging latent immunological changes in the body, eventually resulting in autoimmune demyelination and the appearance of disease symptoms, which confirms the high titer of antibodies to certain viruses in patients with the MS. To first of all herpes simplex virus, Epstein Barr virus, cytomegalovirus and rubella virus. GOAL: Goal of this study is to analyze the incidence of early infection with rubella virus, herpes simplex, cytomegalovirus and Epstein-Barr, in MS patients using titers of IgG and IgM antibodies. MATERIAL AND METHODS: The study included patients treated at the Neurology Clinic in Sarajevo, with a diagnosis of multiple sclerosis (newly discovered) in the period January 2009-December 2011. To all patients beside history and neurological examination and tests to confirm the MS (brain MRI, evoked potentials and CSF examination) made serological tests for viruses, HSV, Rubella virus, cytomegalovirus and Ebstain-Barr's virus, with reference to the previous parameters (old) and new viral infection. RESULTS: In this period there were 118 newly diagnosed multiple sclerosis from which 69.5% (82) female and 30.5% (36) male patients aged 23-56 years. IgG antibodies to herpes simplex virus was positive in 93.2% (110 patients) (72 F and 38 M and IgM only in 0.84% (1 patient). Ig G in Cytomegalovirus was positive in 86.44% (102 subjects, 71 females and 31 males), while IgM was negative in whole sample. IgG Rubella virus was positive in 61.01% (72 patients, 52 F and 20 M) and IgM was negative in all, while IgG in Ebstain-Barr's virus was positive in 83% (98 patients). CONCLUSION: Early infection by herpes simplex virus, cytomegalovirus, Epstein-Barr and Rubella is present in patients with multiple sclerosis in a significant number so the conclusions is the fact that in the development of multiple sclerosis an important role early exposure to these viruses. Key words: early viral infection, multiple sclerosis.

Neoplasms in the clinical material of neurology clinic, Clinical Center of University of Sarajevo in the period 1990-2009.

INTRODUCTION: Brain tumors are a unique and heterogeneous group of tumors with which face a variety of specialties, mostly oncologists, neurologists and neurosurgeons. Due to their specific location all brain tumors are malignant, regardless of their malignant potential, because any expansion process within the skull, increased intracranial pressure and destruction of surrounding structures, which can cause neurological, quantitative disturbances of consciousness or death. GOAL: The goal of this study was to record neoplastic processes of the central nervous system in patients of Neurology Clinic, Clinical Center of Sarajevo University in the twenty-year period (January 1st 1990-December 31st 2009). The study was partly retrospective and partly prospective determined by three time periods. MATERIAL AND METHODS: We reviewed medical records and documentation of patients treated at Neurology Clinic, which has 102 beds. All patients' data were collected using a specially designed questionnaire for this study. RESULTS AND DISCUSSION: The number of secondary tumor process for the period 2000-2005 is greater than in the period 1990-1999, while in the period 2000-2009 is increasing (17.2%-30.3%). The male-female ratio is 52:48. During the first two monitoring period there were statistically significantly more men, and in the last monitoring period there were more women. The mean patient's age was 60 years. The most common symptom was hemiparesis for all observed periods evaluated with standard diagnostic tests: CT and MRI. CONCLUSION: We can conclude that CNS neoplasms in patients of Neurology Clinic, Clinical Center of Sarajevo University are present in the twenty-year period with total of 1.47%, and showed a decrease of 2.7% (1990-1999) to 0.47% for the period 2006-2009.

Betahistine or Cinnarizine for treatment of Meniere's disease.

INTRODUCTION: Meniere's disease is a condition with sudden attacks of vertigo with nausea and vomiting accompanied by loss of hearing and buzzing sensation in the ears, most commonly unilateral. The exact cause of the disease is unknown. Betahistine is the analogue of histamine with weaker agonistic effect on histamine H1 receptors and stronger effect on histamine H3 receptors, while Cinnarizine has more effective effect on H1 receptors. GOAL: The aim is to determine which drug is more effective in the treatment of Meniere's disease Betahistine or Cinnarizine. MATERIAL AND METHODS: This study evaluates the effectiveness of Betahistine in 37 patients with the Meniere's syndrome accompanied by classic triad of symptoms treated in hospital conditions and Cinnarizine effect in 36 patients with a less severe clinical picture, which were treated as outpatients. To all patients were conducted laboratory tests, brain CAT (to exclude possible expansive process, MS or stroke) and TCD in order to eliminate any possible circulatory disturbances in VB basin. Group with classic Meniere's syndrome was treated at a dose of Betahistine of 3 x 16 mg and followed 8 weeks, while the second group was treated with Cinnarizine at a dose of 2 x 75 mg and also followed for 8 weeks. CONCLUSIONS: Already after one month of therapy was noticed better effect in case of Betahistine in terms of symptoms reduction compared to the Cinnarizine effect.

Transcription factor GATA6 promotes migration of human coronary artery smooth muscle cells in vitro.

Vascular smooth muscle cell plasticity plays a pivotal role in the pathophysiology of vascular diseases. Despite compelling evidence demonstrating the importance of transcription factor GATA6 in vascular smooth muscle, the functional role of GATA6 remains poorly understood. The aim of this study was to elucidate the role of GATA6 on cell migration and to gain insight into GATA6-sensitive genes in smooth muscle. We found that overexpression of GATA6 promotes migration of human coronary artery smooth muscle cells in vitro, and that silencing of GATA6 in smooth muscle cells resulted in reduced cellular motility. Furthermore, a complete microarray screen of GATA6-sensitive gene transcription resulted in 739 upregulated and 248 downregulated genes. Pathways enrichment analysis showed involvement of transforming growth factor beta (TGF-ß) signaling which was validated by measuring mRNA expression level of several members. Furthermore, master regulators prediction based on microarray data revealed several members of (mitogen activated protein kinase) MAPK pathway as a master regulators, reflecting involvement of MAPK pathway also. Our findings provide further insights into the functional role of GATA6 in vascular smooth muscle and suggest that targeting GATA6 may constitute as a new approach to inhibit vascular smooth muscle migration.

Inducible Deletion of YAP and TAZ in Adult Mouse Smooth Muscle Causes Rapid and Lethal Colonic Pseudo-Obstruction.

BACKGROUND & AIMS: YAP (Yap1) and TAZ (Wwtr1) are transcriptional co-activators and downstream effectors of the Hippo pathway, which play crucial roles in organ size control and cancer pathogenesis. Genetic deletion of YAP/TAZ has shown their critical importance for embryonic development of the heart, vasculature, and gastrointestinal mesenchyme. The aim of this study was to determine the functional role of YAP/TAZ in adult smooth muscle cells in vivo. METHODS: Because YAP and TAZ are mutually redundant, we used YAP/TAZ double-floxed mice crossed with mice that express tamoxifen-inducible CreERT2 recombinase driven by the smooth muscle-specific myosin heavy chain promoter. RESULTS: Double-knockout of YAP/TAZ in adult smooth muscle causes lethality within 2 weeks, mainly owing to colonic pseudo-obstruction, characterized by severe distension and fecal impaction. RNA sequencing in colon and urinary bladder showed that smooth muscle markers and muscarinic receptors were down-regulated in the YAP/TAZ knockout. The same transcripts also correlated with YAP/TAZ in the human colon. Myograph experiments showed reduced contractility to depolarization by potassium chloride and a nearly abolished muscarinic contraction and spontaneous activity in colon rings of YAP/TAZ knockout. CONCLUSIONS: YAP and TAZ in smooth muscle are guardians of colonic contractility and control expression of contractile proteins and muscarinic receptors. The knockout model has features of human chronic intestinal pseudo-obstruction and may be useful for studying this disease.

MRTFA overexpression promotes conversion of human coronary artery smooth muscle cells into lipid-laden foam cells.

OBJECTIVE: Smooth muscle cells contribute significantly to lipid-laden foam cells in atherosclerotic plaques. However, the underlying mechanisms transforming smooth muscle cells into foam cells are poorly understood. The purpose of this study was to gain insight into the molecular mechanisms regulating smooth muscle foam cell formation. APPROACH AND RESULTS: Using human coronary artery smooth muscle cells we found that the transcriptional co-activator MRTFA promotes lipid accumulation via several mechanisms, including direct transcriptional control of LDL receptor, enhanced fluid-phase pinocytosis and reduced lipid efflux. Inhibition of MRTF activity with CCG1423 and CCG203971 significantly reduced lipid accumulation. Furthermore, we demonstrate enhanced MRTFA expression in vascular remodeling of human vessels. CONCLUSIONS: This study demonstrates a novel role for MRTFA as an important regulator of lipid homeostasis in vascular smooth muscle cells. Thus, MRTFA could potentially be a new therapeutic target for inhibition of vascular lipid accumulation.

Elevated serum homocysteine level is not associated with serum C-reactive protein in patients with probable Alzheimer's disease.

Elevated plasma homocysteine (Hcy) levels have been associated with Alzheimer's disease (AD) and cognitive impairment. Studies have shown that Hcy may have direct and indirect neurotoxicity effects. The aim of the study was to investigate serum Hcy concentration in patients with probable AD with age-matched controls and to determine whether there was an association between serum Hcy and C-reactive protein concentration in patients with probable AD. We also aimed to determine whether there was an association between serum tHcy concentration and cognitive impairment in patients with probable AD. Serum concentration of total Hcy was determined by the fluorescence polarization immunoassay on the AxSYM system, and serum C-reactive protein (CRP) concentration was determined by means of particle-enhanced immunonephelometry with the use of BN II analyzer. Cognitive impairment was tested by the MMSE score. Body mass index (BMI) was calculated for each subject included in the study. Age, systolic and diastolic blood pressure and BMI did not differ significantly between the two groups. Mean serum tHcy concentration in the control group of subjects was 12.60 mumol/L, while in patients with probable AD the mean serum tHcy concentration was significantly higher than 16.15 mumol/L (p < 0.01). A significant negative association between serum tHcy concentration and cognitive impairment tested by the MMSE score in patients with probable AD was determined (r = -0.61634; p < 0.001). Positive, although not significant correlation between CRP and serum tHcy concentrations in patients with AD, was observed. Increased tHcy concentration in patients with probable AD, and the established negative correlation between serum tHcy concentration and cognitive damage tested by MMSE score in the same group of patients, suggests the possible independent role of Hcy in the pathogenesis of AD and cognitive impairment associated with this disease.

Characteristics of symptomatic epilepsy in patients with brain tumours.

The aim of our work is to determine the total number, age, gender of the patients with the symptomatic epileptic seizures associated with brain tumours, tumour location, clinical signs and characteristics of epileptic seizures. We have analyzed medical documentation of the patients with brain tumours hospitalized at the Department of Neurology, University of Sarajevo Clinics Centre. This study is retrospective and includes time period from 1st January 2000 until 31st December 2005. During the observed period at the Department of Neurology in Sarajevo there were in total 9753 hospitalized patients, from which 101 (1,1%) patients with the brain tumour diagnosis. Average patient's age was 62,60 +/- 1,28 years. In one third of the patients (32%) were recorded epileptic seizures, without significant difference between genders. In case of symptomatic epilepsy, significantly more frequent locations of tumours were: in several lobes (28%), parietal lobe (25%), as well as frontal and temporal lobe (18,8% each), while there were no changes in cerebellum and brain stem (chi2 =7,174, p<0,05). The most prominent signs of illness in our sample were hemiparesis with the cranial nerves lesion (56,3%), speech problems (25%). Normal neurologic findings were significantly more frequent among patients with the symptomatic epilepsy (chi2 =6,349, p<0,05). The most often was a single seizure (59%), in 38% of cases there were recorded series of seizures, and only 3% of patients had status epilepticus. In relation to the type of seizures, the most often are simple partial seizures with or without secondary generalization (66%), than generalized convulsive (31%), and the rarest one are complex partial seizures (3%). Symptomatic epilepsy in case of brain tumours occurs in one third of patients, at older age, and in both genders. The lesion usually affects several lobes and cause simple partial seizures with or without secondary generalization. The most often clinical signs in case of all brain tumours are cranial nerves lesion and hemiparesis, while the normal neurologic findings are significantly dominant in the group of patients with the epileptic seizures.