bluuebunny/crossref_metadata_2025 huggingface dataset:
Bluetongue serotype 8 vaccine coverage in northern and south‐eastern England in 2008
Authors: Webb C. R. , Floyd T. , Brien S. , Oura C. A. L. , Wood J. L. N.
Published: April 2011

A postal survey of all registered cattle and sheep farmers in East Anglia was carried out from July 2008 to determine bluetongue virus serotype 8 (BTV‐8) vaccine uptake in the region. The vaccine was available to farmers in this region from May 2008. The survey was repeated in Cumbria and Northumberland at the beginning of 2009. In these regions, the vaccine was not available until September 1, 2008. Holding‐level vaccine uptake was estimated to be 85 per cent (95 per cent confidence interval [CI] 83 to 87 per cent, n=1623) in East Anglia and 36 per cent (95 per cent CI 32 to 40 per cent, n=633) in northern England. A telephone follow‐up of non‐responders reduced these estimates to 79 and 29 per cent in East Anglia and northern England, respectively. In both regions, vaccine coverage was higher in sheep than in cattle, with 92 per cent of sheep in East Anglia having been vaccinated. The proportion of holdings that had applied the vaccine or were intending to apply the vaccine in 2009 in the northern region was 51 per cent (95 per cent CI 47 to 54 per cent, n=664), with a further 37 per cent undecided at the time of response.
Comparison of direct culture versus PCR for the detection of Brucella in aborted fetuses of cattle and sheep in Turkey
Authors: Buyukcangaz E. , Sen A. , Carli K. T. , Kahya S.
Published: April 2011

The aim of this study was to detect Brucella in samples from aborted fetuses of sheep and cattle in Turkey using PCR and bacteriological analysis, and to determine the sensitivity and specificity of the PCR. Organ homogenates from 38 aborted fetuses of cattle and 56 aborted fetuses of sheep were tested. All organ homogenateswere cultured for bacteriological analysis, and all of the homogenates and the Brucella isolates obtained by culture were examined with a commercial PCR kit. On bacteriological analysis, Brucella species were found in 30 (31.9 per cent) of the 94 organ homogenates, eight (21.1 per cent) of which were from cattle and 22 (39.3 per cent) from sheep. Using PCR, a total of 29 (30.9 per cent) homogenates were positive for Brucella species, eight (21.1 per cent) of which were from cattle and 21 (37.5 per cent) from sheep. Compared with the bacteriological method, the diagnostic sensitivity and specificity of the PCR kit used in this study were 83 per cent and 94 per cent, respectively.
Glutathione peroxidase-3 produced by the kidney binds to a population of basement membranes in the gastrointestinal tract and in other tissues
Authors: Burk Raymond F. , Olson Gary E. , Winfrey Virginia P. , Hill Kristina E. , Yin Dengping
Published: April 2011

Glutathione peroxidase-3 (Gpx3), the extracellular glutathione peroxidase synthesized largely in the kidney, binds to basement membranes of renal cortical epithelial cells. The present study assessed extrarenal expression of Gpx3 using RT-PCR and presence of Gpx3 protein using immunocytochemistry. Gpx3 expression was higher in kidney and epididymis than in other tissues. Gpx3 bound to basement membranes of epithelial cells in the gastrointestinal tract, the efferent ducts connecting the seminiferous tubules with the epididymis, the bronchi, and type II pneumocytes. It was not detected on the basement membrane of type I pneumocytes. Gpx3 was also present in the lumen of the epididymis. Transplantation of Gpx3+/+kidneys into Gpx3−/−mice led to Gpx3 binding to the same basement membranes to which it bound in Gpx3+/+mice but not to its presence in the epididymal lumen. These results show that Gpx3 from the blood binds to basement membranes of specific epithelial cells and indicate that the cells modify their basement membranes to cause the binding. They further indicate that at least two Gpx3 compartments exist in the organism. In one compartment, kidney supplies Gpx3 through the blood to specific basement membranes in a number of tissues. In the other compartment, the epididymis provides Gpx3 to its own lumen. Tissues other than kidney and epididymis express Gpx3 at lower levels and may supply Gpx3 to other compartments.
Swim-exercised mice show a decreased level of proteinO-GlcNAcylation and expression ofO-GlcNAc transferase in heart
Authors: Belke Darrell D.
Published: April 2011

Swim-training exercise in mice leads to cardiac remodeling associated with an improvement in contractile function. Protein O-linked N-acetylglucosamine ( O-GlcNAcylation) is a posttranslational modification of serine and threonine residues capable of altering protein-protein interactions affecting gene transcription, cell signaling pathways, and general cell physiology. Increased levels of protein O-GlcNAcylation in the heart have been associated with pathological conditions such as diabetes, ischemia, and hypertrophic heart failure. In contrast, the impact of physiological exercise on protein O-GlcNAcylation in the heart is currently unknown. Swim-training exercise in mice was associated with the development of a physiological hypertrophy characterized by an improvement in contractile function relative to sedentary mice. General protein O-GlcNAcylation was significantly decreased in swim-exercised mice. This effect was mirrored in the level of O-GlcNAcylation of individual proteins such as SP1. The decrease in protein O-GlcNAcylation was associated with a decrease in the expression of O-GlcNAc transferase (OGT) and glutamine-fructose amidotransferase (GFAT) 2 mRNA. O-GlcNAcase (OGA) activity was actually lower in swim-trained than sedentary hearts, suggesting that it did not contribute to the decreased protein O-GlcNAcylation. Thus it appears that exercise-induced physiological hypertrophy is associated with a decrease in protein O-GlcNAcylation, which could potentially contribute to changes in gene expression and other physiological changes associated with exercise.
Responses of neurons in the rostral ventrolateral medulla to whole body rotations: comparisons in decerebrate and conscious cats
Authors: DeStefino V. J. , Reighard D. A. , Sugiyama Y. , Suzuki T. , Cotter L. A. , Larson M. G. , Gandhi N. J. , Barman S. M. , Yates B. J.
Published: April 2011

The responses to vestibular stimulation of brain stem neurons that regulate sympathetic outflow and blood flow have been studied extensively in decerebrate preparations, but not in conscious animals. In the present study, we compared the responses of neurons in the rostral ventrolateral medulla (RVLM), a principal region of the brain stem involved in the regulation of blood pressure, to whole body rotations of conscious and decerebrate cats. In both preparations, RVLM neurons exhibited similar levels of spontaneous activity (median of ∼17 spikes/s). The firing of about half of the RVLM neurons recorded in decerebrate cats was modulated by rotations; these cells were activated by vertical tilts in a variety of directions, with response characteristics suggesting that their labyrinthine inputs originated in otolith organs. The activity of over one-third of RVLM neurons in decerebrate animals was altered by stimulation of baroreceptors; RVLM units with and without baroreceptor signals had similar responses to rotations. In contrast, only 6% of RVLM neurons studied in conscious cats exhibited cardiac-related activity, and the firing of just 1% of the cells was modulated by rotations. These data suggest that the brain stem circuitry mediating vestibulosympathetic reflexes is highly sensitive to changes in body position in space but that the responses to vestibular stimuli of neurons in the pathway are suppressed by higher brain centers in conscious animals. The findings also raise the possibility that autonomic responses to a variety of inputs, including those from the inner ear, could be gated according to behavioral context and attenuated when they are not necessary.
Gambogic Acid Inhibits STAT3 Phosphorylation through Activation of Protein Tyrosine Phosphatase SHP-1: Potential Role in Proliferation and Apoptosis
Authors: Prasad Sahdeo , Pandey Manoj K. , Yadav Vivek R. , Aggarwal Bharat B.
Published: April 2011

The transcription factor, STAT3, is associated with proliferation, survival, and metastasis of cancer cells. We investigated whether gambogic acid (GA), a xanthone derived from the resin of traditional Chinese medicine, Garcinia hanburyi (mangosteen), can regulate the STAT3 pathway, leading to suppression of growth and sensitization of cancer cells. We found that GA induced apoptosis in human multiple myeloma cells that correlated with the inhibition of both constitutive and inducible STAT3 activation. STAT3 phosphorylation at both tyrosine residue 705 and serine residue 727 was inhibited by GA. STAT3 suppression was mediated through the inhibition of activation of the protein tyrosine kinases Janus-activated kinase 1 (JAK1) and JAK2. Treatment with the protein tyrosine phosphatase (PTP) inhibitor pervanadate reversed the GA-induced downregulation of STAT3, suggesting the involvement of a PTP. We also found that GA induced the expression of the PTP SHP-1. Deletion of the SHP-1 gene by siRNA suppressed the ability of GA to inhibit STAT3 activation and to induce apoptosis, suggesting the critical role of SHP-1 in its action. Moreover, GA downregulated the expression of STAT3-regulated antiapoptotic (Bcl-2, Bcl-xL, and Mcl-1), proliferative (cyclin D1), and angiogenic (VEGF) proteins, and this correlated with suppression of proliferation and induction of apoptosis. Overall, these results suggest that GA blocks STAT3 activation, leading to suppression of tumor cell proliferation and induction of apoptosis. Cancer Prev Res; 4(7); 1084–94. ©2011 AACR.
Dietary Fish Oil Promotes Colonic Apoptosis and Mitochondrial Proton Leak in Oxidatively Stressed Mice
Authors: Fan Yang-Yi , Ran Qitao , Toyokuni Shinya , Okazaki Yasumasa , Callaway Evelyn S. , Lupton Joanne R. , Chapkin Robert S.
Published: April 2011

An alteration of mitochondrial function can result in disruption of redox homeostasis and is associated with abnormal cancer cell growth. Manganese superoxide dismutase (SOD2) and glutathione peroxidase 4 (Gpx4) are two of the most important antioxidant defense enzymes that protect cells against oxidative stress. We had previously shown that n-3 polyunsaturated fatty acids (PUFA) promote colonocyte apoptosis, a marker of colon cancer risk, in part by enhancing phospholipid oxidation. To elucidate the mechanisms regulating oxidative stress-induced apoptosis in vivo, we fed heterozygous SOD2Het, Gpx4Het, and transgenic Gpx4Tg mice diets containing either 15% corn oil by weight (CO, enriched in n-6 PUFA) or 3.5% CO + 11.5% fish oil (FO, enriched in n-3 PUFA) for 4 weeks. Our data showed that (i) genetic predeposition to oxidative stress facilitates apoptosis in the mouse colon (Gpx4Het > SOD2Het > Wt > Gpx4Tg), (ii) dietary n-3 PUFA have an additive effect on the induction of apoptosis in Gpx4Het and SOD2Het mice; and (iii) dietary n-3 PUFA reverse the phenotype in oxidatively protected Gpx4Tg mice by elevating apoptosis to a level observed in wild-type (Wt; control) animals. Complimentary experiments examining colonic mitochondrial bioenergetic profiles indicate that FO-fed mice exhibit a significantly (P < 0.05) increased respiration-induced proton leak relative to control CO treatment. This finding was consistent with a loss of membrane potential in response to chronic oxidative stress and supports the contention that n-3 PUFA alter mitochondrial metabolic activity, thereby enhancing apoptosis and reducing colon cancer risk. Cancer Prev Res; 4(8); 1267–74. ©2011 AACR.
Accurate Reconstruction of the Temporal Order of Mutations in Neoplastic Progression
Authors: Sprouffske Kathleen , Pepper John W. , Maley Carlo C.
Published: April 2011

The canonical route from normal tissue to cancer occurs through sequential acquisition of somatic mutations. Many studies have constructed a linear genetic model for tumorigenesis using the genetic alterations associated with samples at different stages of neoplastic progression from cross-sectional data. The common interpretation of these models is that they reflect the temporal order within any given tumor. Linear genetic methods implicitly neglect genetic heterogeneity within a neoplasm; each neoplasm is assumed to consist of one dominant clone. We modeled neoplastic progression of colorectal cancer using an agent-based model of a colon crypt and found clonal heterogeneity within our simulated neoplasms, as observed in vivo. Just 7.3% of cells within neoplasms acquired mutations in the same order as the linear model. In 41% of the simulated neoplasms, no cells acquired mutations in the same order as the linear model. We obtained similarly poor results when comparing the temporal order with oncogenetic tree models inferred from cross-sectional data. However, when we reconstructed the cell lineage of mutations within a neoplasm using several biopsies, we found that 99.7% cells within neoplasms acquired their mutations in an order consistent with the cell lineage mutational order. Thus, we find that using cross-sectional data to infer mutational order is misleading, whereas phylogenetic methods based on sampling intratumor heterogeneity accurately reconstructs the evolutionary history of tumors. In addition, we find evidence that disruption of differentiation is likely the first lesion in progression for most cancers and should be one of the few regularities of neoplastic progression across cancers. Cancer Prev Res; 4(7); 1135–44. ©2011 AACR.
Protective and Restorative Effects of Magnolol on Neurotoxicity in Mice with 6-Hydroxydopamine-Induced Hemiparkinsonism
Authors: Chen Hwei-Hsien , Lin Shiau-Chin , Chan Ming-Huan
Published: April 2011

Parkinson’s disease (PD) is one of the most common neurodegenerative disorders. The aim of the present study was to investigate the protective and restorative potential of magnolol, a major bioactive biphenolic from the bark of Magnolia officinalis, for alleviating the motor deficits induced by 6-hydroxydopamine (6-OHDA) in a mouse model of PD. Before or after unilateral striatal 6-OHDA lesion induction, mice were administered magnolol subchronically; then the apomorphine-induced rotational behaviors of the hemiparkinsonian mice and tyrosine hydroxylase (TH) expression in striatum were determined. Magnolol that was administered 30 min before 6-OHDA lesion induction and then applied daily for 14 days significantly ameliorated apomorphine-induced contralateral rotation in 6-OHDA-lesioned mice, and consistently protected the decreased levels of TH protein expression in striatum. One week after termination of the 7-day subchronic pretreatment, magnolol also remarkably prevented the dopaminergic neuronal loss as identified by TH immunohistochemistry staining in striatum, associated with rotational behavioral protection in 6-OHDA-lesioned mice. Importantly, daily subchronic posttreatment with magnolol for 14 days efficiently reduced apomorphine-induced rotation, but did not restore the neuronal impairment in striatum damaged by 6-OHDA. Taken together, these findings suggest that magnolol may possess neuronal protective activity and behavioral restoration against 6-OHDA-induced toxicity in the PD model.
Sleep Disorders in Spinocerebellar Ataxia Type 2 Patients
Authors: Velázquez-Pérez Luis , Voss Ursula , Rodríguez-Labrada Roberto , Auburger Georg , Canales Ochoa Nalia , Sánchez Cruz Gilberto , Galicia Polo Lourdes , Haro Valencia Reyes , Aguilera Rodríguez Raúl , Medrano Montero Jacqueline , Laffita Mesa Jose M. , Tuin Inka
Published: April 2011

Background: Sleep disturbances are common features in spinocerebellar ataxias (SCAs). Nevertheless, sleep data on SCA2 come from scarce studies including few patients, limiting the evaluation of the prevalence and determinants of sleep disorders. Objective: To assess the frequency and possible determinants of sleep disorders in the large and homogeneous SCA2 Cuban population. Methods: Thirty-two SCA2 patients and their age- and sex-matched controls were studied by video-polysomnography and sleep interviews. Results: The most striking video-polysomnography features were rapid eye movement (REM) sleep pathology and periodic leg movements (PLMs). REM sleep abnormalities included a consistent reduction of the REM sleep percentage and REM density as well as an increase in REM sleep without atonia (RWA). REM sleep and REM density decreases were closely related to the increase in ataxia scores, whereas the RWA percentage was influenced by the cytosine-adenine-guanine (CAG) repeats. PLMs were observed in 37.5% of cases. The PLM index showed a significant association with the ataxia score and disease duration but not with CAG repeats. Conclusions: REM sleep pathology and PLMs are closely related to SCA2 severity, suggesting their usefulness as disease progression markers. The RWA percentage is influenced by the CAG repeats and might thus be a sensitive parameter for reflecting polyglutamine toxicity. Finally, as PLMs are sensible to drug treatment, they represents a new therapeutic target for the symptomatic treatment of SCA2.
Bilateral Subthalamic Deep Brain Stimulation for Camptocormia Associated with Parkinson’s Disease
Authors: Asahi Takashi , Taguchi Yoshiharu , Hayashi Nakamasa , Hamada Hideo , Dougu Nobuhiro , Takashima Shutaro , Tanaka Kortaro , Endo Shunro
Published: April 2011

Background: Few multiple case studies of the effects of deep brain stimulation for camptocormia associated with Parkinson’s disease have been reported. Although deep brain stimulation was in some cases not effective against camptocormia, it is unclear in which types of patients it was effective in treating camptocormia. Objective: We treated 4 Parkinson’s disease patients with camptocormia and evaluated their paraspinal muscle status by computed tomography to specify the characteristics of cases of effective treatment. Methods: The 2 female and 2 male patients in this study were 60–69 years old, with a disease duration from onset to surgery of 7–13 years and a follow-up period of 18–40 months. The electrodes were implanted bilaterally in the subthalamic nuclei. Results: Camptocormia was improved in 3 cases, and was unchanged in the remaining case although other parkinsonian symptoms improved. The computed tomography number of paraspinal muscle in the unimproved patient was much smaller than that in the improved patients. Conclusions: A relationship may exist between improvement of camptocormia and severity of paraspinal muscle degeneration.
Dynamic Medium Containing Kit Ligand and Follicle-Stimulating Hormone Promotes Follicular Survival, Activation, and Growth during Long-Term in vitro Culture of Caprine Preantral Follicles
Authors: Lima I.M.T. , Celestino J.J.H. , Faustino L.R. , Magalhães-Padilha D.M. , Rossetto R. , Brito I.R. , Donato M.A.M. , Lopes C.A.P. , Campello C.C. , Peixoto C.A. , Figueiredo J.R. , Rodrigues A.P.R.
Published: April 2011

The aim of this study was to evaluate the effects of a dynamic medium containing kit ligand (KL) and follicle-stimulating hormone (FSH) on the in vitro culture of caprine preantral follicles for 16 days. Ovarian fragments were cultured in α-MEM+ containing or not containing KL (50 ng/ml) and/or FSH (50 ng/ml) added during the first (days 0–8) and/or second half (days 8–16) of the culture period. Noncultured (control) and cultured fragments were processed for histological and ultrastructural evaluation. After 1 day of culture, only the treatments performed with KL or FSH maintained a percentage of normal follicles similar to that of the control. After 16 days, all treatments using KL until day 8 (KL/KL, KL/FSH, and KL/FSH+KL) and only FSH during the entire culture period (FSH/FSH) showed higher rates of follicular survival compared to α-MEM+ alone. After 1 and 8 days, the treatments initially cultured with KL increased the percentage of follicular activation in comparison to α-MEM+ alone and other treatments. The highest follicular diameter after 16 days was observed in follicles cultured with KL until day 8 followed by FSH (KL/FSH). Furthermore, this treatment promoted, as early as after 1 day of culture, an increase in oocyte growth compared to α-MEM+ alone. Ultrastructural analysis confirmed the integrity of follicles cultured in KL/FSH after 16 days. In conclusion, a dynamic medium containing KL and FSH maintained follicular integrity and promoted follicular activation and growth during the long-term in vitro culture of caprine preantral follicles.
Tetramethylpyrazine Inhibits Hypoxia-Induced Pulmonary Vascular Leakage in Rats via the ROS-HIF-VEGF Pathway
Authors: Zhang Le , Deng Mengyang , Zhou Shiwen
Published: April 2011

Tetramethylpyrazine (TMP) is a reactive oxygen species (ROS) antagonist that has potent properties for the treatment of a variety of vascular diseases, such as ischemic stroke and pulmonary hypertension secondary to chronic obstructive pulmonary diseases. However, there are few data about the role of TMP in hypoxia-induced pulmonary vascular leakage. This study examined the effect of TMP on hypoxia-induced pulmonary vascular leakage and the underlying mechanisms. Rat pulmonary microvascular endothelial cells (RPMVECs) treated with TMP or not were subjected to hypoxic or normoxic conditions for 24 h, and the monolayer permeability, intracellular ROS, hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) proteins levels were determined. Additionally, rats administrated TMP were exposed to hypobaric hypoxia to evaluate the effect of TMP in vivo by measuring lung water content, pulmonary vascular leakage into the lungs and immunohistochemistry for HIF-1α and VEGF. Hypoxia was found to cause a significant increase in RPMVEC monolayer permeability and intracellular ROS, HIF-1α and VEGF protein levels. Treatment with TMP decreased the hypoxia-induced RPMVEC monolayer permeability and attenuated the elevation of ROS, HIF-1α and VEGF protein levels. TMP-treated animals showed less pulmonary vascular leakage and HIF-1α and VEGF expression compared with those exposed to hypoxia alone. These observations supported that TMP inhibited the increase in pulmonary vascular permeability induced by hypoxia. The underlying mechanisms may be related to the scavenging of intracellular ROS and the suppression of hypoxia-induced upregulation of HIF-1α and VEGF proteins.
Exposure to Prenatal Psychobiological Stress Exerts Programming Influences on the Mother and Her Fetus
Authors: Sandman Curt A. , Davis Elysia P. , Buss Claudia , Glynn Laura M.
Published: April 2011

Background/Aims: Accumulating evidence from a relatively small number of prospective studies indicates that exposure to prenatal stress profoundly influences the developing human fetus with consequences that persist into childhood and very likely forever. Methods: Maternal/fetal dyads are assessed at ∼20, ∼25, ∼31 and ∼36 weeks of gestation. Infant assessments begin 24 h after delivery with the collection of cortisol and behavioral responses to the painful stress of the heel-stick procedure and measures of neonatal neuromuscular maturity. Infant cognitive, neuromotor development, stress and emotional regulation are evaluated at 3, 6 12 and 24 months of age. Maternal psychosocial stress and demographic information is collected in parallel with infant assessments. Child neurodevelopment is assessed with cognitive tests, measures of adjustment and brain imaging between 5 and 8 years of age. Results:Psychobiological markers of stress during pregnancy, especially early in gestation, result in delayed fetal maturation, disrupted emotional regulation and impaired cognitive performance during infancy and decreased brain volume in areas associated with learning and memory in 6- to 8-year-old children. We review findings from our projects that maternal endocrine alterations that accompany pregnancy and influence fetal/infant/child development are associated with decreased affective responses to stress, altered memory function and increased risk for postpartum depression. Conclusions: Our findings indicate that the mother and her fetus both are influenced by exposure to psychosocial and biological stress. The findings that fetal and maternal programming occur in parallel may have important implications for long-term child development and mother/child interactions.
Estimated Glomerular Filtration Rate in Short-Risk Stratification in Acute Myocardial Infarction
Authors: Blasco Luisa , Sanjuan Rafael , Carbonell Nieves , Solís Miguel A. , Puchades María J. , Torregrosa Isidro , Miguel Juan A.
Published: April 2011

Background/Aims: Renal dysfunction is associated with a higher risk of cardiovascular disease in patients with acute myocardial infarction (AMI). The aim of this study was to investigate the independent prognostic value of renal dysfunction and its incremental predictability risk after adjusting for well-known clinical factors in patients with AMI. Methods: 751 consecutive patients with AMI admitted to the Coronary Care Unit (CCU) were included. Patients were grouped into 2 categories according to the baseline estimated glomerular filtration rate (eGFR) on admission (eGFR <60 vs. eGFR ≧60 ml/min/1.73 m2). C-reactive protein and white blood cell count (WBC) as well as clinical prognostic variables were assessed. The endpoint was mortality during CCU stay. The discriminatory power was estimated by the C-index. Results: The patient group with an eGFR <60 ml/min/1.73 m2 was older, had more cardiovascular risk factors, a lower left ventricular ejection fraction and higher cardiovascular mortality during CCU stay (13 vs. 3%). Logistic regression analysis revealed the following predictors of mortality: degree of renal impairment (eGFR <60 ml/min/1.73 m2), hazard ratio (HR) = 2.2 (95% CI 1.1–4.3; p = 0.028); WBC >11,000 × 106/l, HR = 2.3 (95% CI 1.2–4.5; p = 0.017); Killip class on admission, HR = 3.8 (95% CI 1.7–8.5; p = 0.001), and New York Heart Association Functional Classification, HR = 3.6 (95% CI 1.7–7.4; p = 0.001). The adjusted C-index was 0.78 for baseline clinical variables and 0.84 for eGFR. Conclusions: In patients with AMI, decreased eGFR is an important prognostic factor for impaired cardiac function and mortality in the short-term follow-up. The eGFR may be reliably used in the risk stratification of patients with AMI.
An Emerging Role for Understanding Orthostatic Hyp‘er’tension in the Cardiorenal Syndrome
Authors: Buddineni Jaya P. , Chauhan Laxmi , Ahsan Syed T. , Whaley-Connell Adam
Published: April 2011

Orthostatic hypertension (OHT) is a clinically important problem increasingly recognized in persons with borderline hypertension, diabetes mellitus, and autonomic neuropathies, and in the elderly. Moreover, the association of OHT with progression of target end-organ damage, especially coronary heart disease and chronic kidney disease (CKD), and the attendant increased cardiovascular disease (CVD) and CKD risk, is gaining attention but is still underappreciated. There are various mechanisms that contribute to the development of OHT: excessive vascular adrenergic sensitivity, baroreceptor reflex abnormalities, and inappropriate activation of the renin-angiotensin-aldosterone system, which are also mechanisms that lead to cardiorenal metabolic disease (CRS). While the evidence is compelling for the clinical importance of OHT, more investigation is needed to evaluate the effects of OHT on CKD and CVD. The notion that the development of OHT is a risk factor for the development of CRS raises the need for further clinical and investigational attention to this clinical dilemma.
Prevalence and Predictors of Carotid Wall Triple Line Pattern in a General Population Sample
Authors: Ryabikov Andrew , Malyutina Sofia , Halcox Julian , Nikitin Yuri , Marmot Michael , Bobak Martin
Published: April 2011

Objective—

Carotid intima-media thickness (IMT) and plaques are markers of atherosclerosis and predict cardiovascular events. A specific sonographic triple line pattern (TLP) of the carotid wall has been identified in different conditions, but its origin and clinical significance are unclear. We examined the prevalence and predictors of TLP in a general population.

Methods and Results—

The study was conducted in random sample of the general population of Novosibirsk, Russia, within the international Health, Alcohol and Psychosocial Factors in Eastern Europe project. In a subsample of 418 men (aged 45 to 69), carotid IMT, the presence of atherosclerotic plaques, and the presence of TLP were assessed by ultrasound. The prevalence of TLP was 21%. It was associated with IMT (odds ratio=9.53 per 1 SD,
P
<0.001) and the presence of plaques (odds ratio=2.42,
P
=0.002). Other predictors of TLP in multivariate models included age, systolic blood pressure, total cholesterol, and smoking. In addition, infrequent consumption of high amounts of alcohol approximately doubled the risk of triple pattern.

Conclusion—

Our findings showed high prevalence of TLP of carotid wall in a general male population sample from a typical Russian city. This sonographic pattern was strongly associated with cardiovascular risk factors and diseases, bioimaging indicators of atherosclerosis, and episodic heavy drinking.
KCNE5
(
KCNE1L
) Variants Are Novel Modulators of Brugada Syndrome and Idiopathic Ventricular Fibrillation
Authors: Ohno Seiko , Zankov Dimitar P. , Ding Wei-Guang , Itoh Hideki , Makiyama Takeru , Doi Takahiro , Shizuta Satoshi , Hattori Tetsuhisa , Miyamoto Akashi , Naiki Nobu , Hancox Jules C. , Matsuura Hiroshi , Horie Minoru
Published: April 2011

Background—

Brugada syndrome (BrS) has a significantly higher incidence among the male sex. Among genes coding ion channels and their modulatory proteins,
KCNE5
(
KCNE1L
) is located in the X chromosome and encodes an auxiliary β-subunit for K channels. KCNE5 has been shown to modify the transient outward current (I
to
), which plays a key role in determining the repolarization process in the myocardium. This study investigated whether
KCNE5
mutations could be responsible for BrS and other idiopathic ventricular fibrillation (IVF).

Methods and Results—

In 205 Japanese patients with BrS or IVF who tested negative for
SCN5A
mutation, we conducted a genetic screen for
KCNE5
variants. We identified 2 novel
KCNE5
variants: p.Y81H in 3 probands and p.[D92E;E93X] in 1 proband from 4 unrelated families. Y81H was identified in 1 man and 2 women; D92E;E93X was found in a 59-year-old man. All probands received implantable cardioverter-defibrillators. Functional consequences of the
KCNE5
variants were determined through biophysical assay using cotransfection with
KCND3
or
KCNQ1
. In the experiments with
KCND3
, which encodes Kv4.3, I
to
was significantly increased for both
KCNE5
variants compared to wild type. In contrast, there were no significant changes in current properties reconstructed by
KCNQ1
+ wild type
KCNE5
and the 2 variants. With the simulation model, both variants demonstrated notch-and-dome or loss-of-dome patterns.

Conclusions—

KCNE5 modulates I
to
, and its novel variants appeared to cause IVF, especially BrS, in male patients through gain-of-function effects on I
to
. Screening for
KCNE5
variants is relevant for BrS or IVF.
Gaps in the Ablation Line as a Potential Cause of Recovery From Electrical Isolation and Their Visualization Using MRI
Authors: Ranjan Ravi , Kato Ritsushi , Zviman Menekhem M. , Dickfeld Timm M. , Roguin Ariel , Berger Ronald D. , Tomaselli Gordon F. , Halperin Henry R.
Published: April 2011

Background—

Ablation has become an important tool in treating atrial fibrillation and ventricular tachycardia, yet the recurrence rates remain high. It is well established that ablation lines can be discontinuous and that conduction through the gaps in ablation lines can be affected by tissue heating. In this study, we looked at the effect of tissue conductivity and propagation of electric wave fronts across ablation lines with gaps, using both simulations and an animal model.

Methods and Results—

For the simulations, we implemented a 2-dimensional bidomain model of the cardiac syncytium, simulating ablation lines with gaps of varying lengths, conductivity, and orientation. For the animal model, transmural ablation lines with a gap were created in 7 mongrel dogs. The gap length was progressively decreased until there was conduction block. The ablation line with a gap was then imaged using MRI and was correlated with histology. With normal conductivity in the gap and the ablation line oriented parallel to the fiber direction, the simulation predicted that the maximum gap length that exhibited conduction block was 1.4 mm. As the conductivity was decreased, the maximum gap length with conduction block increased substantially, that is, with a conductivity of 67% of normal, the maximum gap length with conduction block increased to 4 mm. In the canine studies, the maximum gap length that displayed conduction block acutely as measured by gross pathology correlated well (
R
2
of 0.81) with that measured by MRI.

Conclusions—

Conduction block can occur across discontinuous ablation lines. Moreover, with recovery of conductivity over time, ablation lines with large gaps exhibiting acute conduction block may recover propagation in the gap over time, allowing recurrences of arrhythmias. The ability to see gaps acutely using MRI will allow for targeting these sites for ablation.
Thoracoscopic Video-Assisted Pulmonary Vein Antrum Isolation, Ganglionated Plexus Ablation, and Periprocedural Confirmation of Ablation Lesions
Authors: Krul Sébastien P.J. , Driessen Antoine H.G. , van Boven Wim J. , Linnenbank André C. , Geuzebroek Guillaume S.C. , Jackman Warren M. , Wilde Arthur A.M. , de Bakker Jacques M.T. , de Groot Joris R.
Published: April 2011

Background—

Thoracoscopic pulmonary vein isolation (PVI) and ganglionated plexus ablation is a novel approach in the treatment of atrial fibrillation (AF). We hypothesize that meticulous electrophysiological confirmation of PVI results in fewer recurrences of AF during follow-up.

Methods and Results—

Surgery was performed through 3 ports bilaterally. Ganglionated plexi were localized and subsequently ablated. PVI was performed and entry and exit block was confirmed. Additional left atrial ablation lines were created and conduction block verified in patients with nonparoxysmal AF. The left atrial appendage was removed. Freedom of AF was assessed by ECGs and Holter monitoring every 3 months or during symptoms of arrhythmia. Antiarrhythmic drugs were discontinued after 3 months and oral anticoagulants were discontinued according to the guidelines. Thirty-one patients were treated (16 paroxysmal AF, 13 persistent AF, 2 long-standing persistent AF). Thirteen patients with nonparoxysmal received additional left atrial ablation lines. After 1 year, 19 of 22 patients (86%) had no recurrences of AF, atrial flutter, or atrial tachycardia and were not using antiarrhythmic drugs (11/12 paroxysmal, 7/9 persistent, and 1/1 long-standing persistent). Three patients had a sternotomy because of uncontrolled bleeding during thoracoscopic surgery. Four adverse events were 1 hemothorax, 1 pneumothorax, and 2 pneumonia. No thromboembolic complications or mortality occurred.

Conclusions—

Thoracoscopic surgery with PVI and ganglionated plexus ablation for AF is a safe and successful procedure with a single procedure success rate of 86% at 1 year. Electrophysiological guided thorough PVI and additional left atrial ablation line creation presumably contributes in achieving a high success rate in the surgical treatment of AF.
A Novel
KCNJ2
Nonsense Mutation, S369X, Impedes Trafficking and Causes a Limited Form of Andersen-Tawil Syndrome
Authors: Doi Takahiro , Makiyama Takeru , Morimoto Takeshi , Haruna Yoshisumi , Tsuji Keiko , Ohno Seiko , Akao Masaharu , Takahashi Yoshiaki , Kimura Takeshi , Horie Minoru
Published: April 2011

Background—

Mutations in
KCNJ2
, a gene encoding the inward rectifier K
+
channel Kir2.1, are associated with Andersen-Tawil syndrome (ATS), which is characterized by (1) ventricular tachyarrhythmias associated with QT (QU)-interval prolongation, (2) periodic paralysis, and (3) dysmorphic features.

Methods and Results—

We identified a novel
KCNJ2
mutation, S369X, in a 13-year-old boy with prominent QU-interval prolongation and mild periodic paralysis. The mutation results in the truncation at the middle of the cytoplasmic C-terminal domain that eliminates the endoplasmic reticulum (ER)-to-Golgi export signal. Current recordings from Chinese hamster ovary cells transfected with
KCNJ2
-S369X exhibited significantly smaller K
+
currents compared with
KCNJ2
wild type (WT) (1 μg each) (−84±14 versus −542±46 picoamperes per picofarad [pA/pF]; −140 mV;
P
<0.0001). Coexpression of the WT and S369X subunits did not show a dominant-negative suppression effect but yielded larger currents than those of WT+S369X (−724±98 pA/pF>−[84+542] pA/pF; 1 μg each; −140 mV). Confocal microscopy analysis showed that the fluorescent protein-tagged S369X subunits were predominantly retained in the ER when expressed alone; however, the expression of S369X subunits to the plasma membrane was partially restored when coexpressed with WT. Fluorescence resonance energy transfer analysis demonstrated direct protein-protein interactions between WT and S369X subunits in the intracellular compartment.

Conclusions—

The S369X mutation causes a loss of the ER export motif. However, the trafficking deficiency can be partially rescued by directly assembling with the WT protein, resulting in a limited restoration of plasma membrane localization and channel function. This alleviation may explain why our patient presented with a relatively mild ATS phenotype.
Genetic and Clinical Correlates of Early-Outgrowth Colony-Forming Units
Authors: Shaw Stanley Y. , Cheng Susan , Cupples L. Adrienne , Larson Martin G. , McCabe Elizabeth L. , Ngwa Julius S. , Wang Ying A. , Martin Roderick P. , Klein Rachael J. , Hashmi Basma , Ajijola Olujimi A. , Lau Evan , O'Donnell Christopher J. , Vasan Ramachandran S. , Cohen Kenneth S. , Wang Thomas J.
Published: April 2011

Background—

Several bone marrow–derived cell populations may have angiogenic activity, including cells termed endothelial progenitor cells. Decreased numbers of circulating angiogenic cell populations have been associated with increased cardiovascular risk. However, few data exist from large, unselected samples, and the genetic determinants of these traits are unclear.

Methods and Results—

We examined the clinical and genetic correlates of early-outgrowth colony-forming units (CFUs) in 1799 participants of the Framingham Heart Study (mean age, 66 years; 54% women). Among individuals without cardiovascular disease (n=1612), CFU number was inversely related to advanced age (
P
=0.004), female sex (
P
=0.04), and triglycerides (
P
=0.008) and positively related to hormone replacement (
P
=0.008) and statin therapy (
P
=0.027) in stepwise multivariable analyses. Overall, CFU number was inversely related to the Framingham risk score (
P
=0.01) but not with prevalent cardiovascular disease. In genome-wide association analyses in the entire sample, polymorphisms were associated with CFUs at the
MOSC1
locus (
P
=3.3×10
−7
) and at the
SLC22A3-LPAL2-LPA
locus (
P
=4.9×10
−7
), a previously replicated susceptibility locus for myocardial infarction. Furthermore, alleles at the
SLC22A3-LPAL2-LPA
locus that were associated with decreased CFUs were also related to increased risk of myocardial infarction (
P
=1.1×10
−4
).

Conclusions—

In a community-based sample, early-outgrowth CFUs are inversely associated with select cardiovascular risk factors. Furthermore, genetic variants at the
SLC22A3-LPAL2-LPA
locus are associated with both decreased CFUs and an increased risk of myocardial infarction. These findings are consistent with the hypothesis that decreased circulating angiogenic cell populations promote susceptibility to myocardial infarction.
Coronary Artery Endothelial Transcriptome In Vivo
Authors: Civelek Mete , Manduchi Elisabetta , Riley Rebecca J. , Stoeckert Christian J. , Davies Peter F.
Published: April 2011

Background—

Endothelial function is central to the localization of atherosclerosis. The in vivo endothelial phenotypic footprints of arterial bed identity and site-specific atherosusceptibility are addressed.

Methods and Results—

Ninety-eight endothelial cell samples from 13 discrete coronary and noncoronary arterial regions of varying susceptibilities to atherosclerosis were isolated from 76 normal swine. Transcript profiles were analyzed to determine the steady-state in vivo endothelial phenotypes. An unsupervised systems biology approach using weighted gene coexpression networks showed highly correlated endothelial genes. Connectivity network analysis identified 19 gene modules, 12 of which showed significant association with circulatory bed classification. Differential expression of 1300 genes between coronary and noncoronary artery endothelium suggested distinct coronary endothelial phenotypes, with highest significance expressed in gene modules enriched for biological functions related to endoplasmic reticulum (ER) stress and unfolded protein binding, regulation of transcription and translation, and redox homeostasis. Furthermore, within coronary arteries, comparison of endothelial transcript profiles of susceptible proximal regions to protected distal regions suggested the presence of ER stress conditions in susceptible sites. Accumulation of reactive oxygen species throughout coronary endothelium was greater than in noncoronary endothelium consistent with coronary artery ER stress and lower endothelial expression of antioxidant genes in coronary arteries.

Conclusions—

Gene connectivity analyses discriminated between coronary and noncoronary endothelial transcript profiles and identified differential transcript levels associated with increased ER and oxidative stress in coronary arteries consistent with enhanced susceptibility to atherosclerosis.
Malignant Tumor Formation After Transplantation of Short-Term Cultured Bone Marrow Mesenchymal Stem Cells in Experimental Myocardial Infarction and Diabetic Neuropathy
Authors: Jeong Jin-Ok , Han Ji Woong , Kim Jin-Man , Cho Hyun-Jai , Park Changwon , Lee Namho , Kim Dong-Wook , Yoon Young-Sup
Published: April 2011

Rationale:

Bone marrow (BM)–derived mesenchymal stem cells (MSCs) hold great promise for cardiovascular cell therapy owing to their multipotency and culture expandability.

Objective:

The aim of the study was to investigate whether MSCs can treat experimental acute myocardial infarction (MI) and diabetic neuropathy.

Methods and Results:

We isolated mononuclear cells from mouse BM and cultured MSCs in a conventional manner. Flow cytometry analyses of these cultured cells at passage 4 showed expression of typical MSC markers such as CD44 and CD29, but not hematopoietic markers such as c-kit, flk1, and CD34. To determine the therapeutic effects of MSCs, we injected MSCs into the peri-infarct area after ligation of the left anterior descending coronary arteries of mice and, as separate experiments, injected the same batch of MSCs into hindlimb muscles of mice with diabetic neuropathy. During the follow-up at 4 to 8 weeks after cell transplantation, growing tumors were observed in 30% of hearts in the MI model, and in 46% of hindlimbs in the diabetic neuropathy model. Histological examination of the tumors revealed hypercelluarity, pleomorphic nucleoli, cytological atypia and necrosis, and positive staining for α-smooth muscle actin, indicative of malignant sarcoma with myogenic differentiation. Chromosomal analysis of these MSCs showed multiple chromosomal aberrations including fusion, fragmentation, and ring formation.

Conclusions:

Genetically unmodified MSCs can undergo chromosomal abnormalities even at early passages and form malignant tumors when transplanted in vivo. These results suggest that careful monitoring of chromosomal status is warranted when in vitro expanded MSCs are used for cell therapy such as for MI.
Activation of Endothelial Toll-Like Receptor 3 Impairs Endothelial Function
Authors: Zimmer Sebastian , Steinmetz Martin , Asdonk Tobias , Motz Inga , Coch Christoph , Hartmann Evelyn , Barchet Winfried , Wassmann Sven , Hartmann Gunther , Nickenig Georg
Published: April 2011

Rationale:

Endothelial dysfunction and atherosclerosis are chronic inflammatory diseases characterized by activation of the innate and acquired immune system. Specialized protein receptors of the innate immune system recognize products of microorganisms and endogenous ligands such as nucleic acids. Toll-like receptor 3 (TLR3), for example, detects long double-stranded RNA and is abundantly expressed in endothelial cells. Whether innate immunity contributes to atherogenic mechanisms in endothelial cells is poorly understood.

Objective:

We sought to determine the effects of TLR3 activation in endothelial cells.

Methods and Results:

We first investigated whether stimulation of TLR3 influences endothelial biology in mice. Intravenous injection of polyinosine polycytidylic acid, a synthetic double-stranded RNA analog and TLR3 ligand, impaired endothelium-dependent vasodilation, increased vascular production of reactive oxygen species, and reduced reendothelialization after carotid artery injury in wild-type mice compared with controls but had no effect in TLR3
−/−
animals. TLR3 stimulation not only induced endothelial dysfunction but also enhanced the formation of atherosclerotic plaques in apolipoprotein E–deficient mice. In vitro incubation of endothelial cells with polyinosine polycytidylic acid induced production of the proinflammatory cytokines interleukin-8 and interferon-γ–induced protein 10, increased formation of reactive oxygen species, diminished proliferation, and increased apoptosis, which suggests that endothelial cells are able to directly detect and respond to TLR3 ligands. Neutralization of interleukin-8 and interferon-γ–induced protein 10 antagonizes the observed negative effects of polyinosine polycytidylic acid. We found elevated levels of circulating endothelial progenitor cells in polyinosine polycytidylic acid–treated mice, although they displayed increased endothelial dysfunction. Stimulation of TLR3 in cultured endothelial progenitor cells, however, led to increased formation of reactive oxygen species, increased apoptosis, and reduced migration. Injection of endothelial progenitor cells that had been incubated with polyinosine polycytidylic acid ex vivo hindered reendothelialization after carotid artery injury. Therefore, endothelial progenitor cell function was affected by TLR3 stimulation. Finally, apolipoprotein E–deficient/TLR3-deficient mice exhibited improved endothelial function compared with apolipoprotein E–deficient/TLR3
+/+
littermates.

Conclusions:

Immunorecognition of long double-stranded RNA by endothelial cells may be an important mechanism involved in endothelial cell activation and development of endothelial dysfunction.