bluuebunny/crossref_metadata_2025 huggingface dataset:
Massive weight loss-induced mechanical plasticity in obese gait
Authors: Hortobágyi Tibor , Herring Cortney , Pories Walter J. , Rider Patrick , DeVita Paul
Published: August 2011

We examined the hypothesis that metabolic surgery-induced massive weight loss causes mass-driven and behavioral adaptations in the kinematics and kinetics of obese gait. Gait analyses were performed at three time points over ∼1 yr in initially morbidly obese (mass: 125.7 kg; body mass index: 43.2 kg/m2) but otherwise healthy adults. Ten obese adults lost 27.1% ± 5.1 (34.0 ± 9.4 kg) weight by the first follow-up at 7.0 mo (±0.7) and 6.5 ± 4.2% (8.2 ± 6.0 kg) more by the second follow-up at 12.8 mo (±0.9), with a total weight loss of 33.6 ± 8.1% (42.2 ± 14.1 kg; P = 0.001). Subjects walked at a self-selected and a standard 1.5 m/s speed at the three time points and were also compared with an age- and gender-matched comparison group at the second follow-up. Weight loss increased swing time, stride length, gait speed, hip range of motion, maximal knee flexion, and ankle plantarflexion. Weight loss of 27% led to 3.9% increase in gait speed. An additional 6.5% weight loss led to an additional 7.3% increase in gait speed. Sagittal plane normalized knee torque increased and absolute ankle and frontal plane knee torques decreased after weight loss. We conclude that large weight loss produced mechanical plasticity by modifying ankle and knee torques and gait behavior. There may be a weight loss threshold of 30 kg limiting changes in gait kinematics. Implications for exercise prescription are also discussed.
Effects of acute sprint interval cycling and energy replacement on postprandial lipemia
Authors: Freese Eric C. , Levine Ari S. , Chapman Donald P. , Hausman Dorothy B. , Cureton Kirk J.
Published: August 2011

High postprandial blood triglyceride (TG) levels increase cardiovascular disease risk. Exercise interventions may be effective in reducing postprandial blood TG. The purpose of this study was to determine the effects of sprint interval cycling (SIC), with and without replacement of the energy deficit, on postprandial lipemia. In a repeated-measures crossover design, six men and six women participated in three trials, each taking place over 2 days. On the evening of the first day of each trial, the participants either did SIC without replacing the energy deficit (Ex-Def), did SIC and replaced the energy deficit (Ex-Bal), or did not exercise (control). SIC was performed on a cycle ergometer and involved four 30-s all-out sprints with 4-min active recovery. In the morning of day 2, responses to a high-fat meal were measured. Venous blood samples were collected in the fasted state and at 0, 30, 60, 120, and 180 min postprandial. There was a trend toward a reduction with treatment in fasting TG ( P = 0.068), but no significant treatment effect for fasting insulin, glucose, nonesterified fatty acids, or betahydroxybutryrate ( P > 0.05). The postprandial area under the curve (mmol·l−1·3 h−1) TG response was significantly lower in Ex-Def (21%, P = 0.006) and Ex-Bal (10%, P = 0.044) than in control, and significantly lower in Ex-Def (12%, P = 0.032) than in Ex-Bal. There was no treatment effect ( P > 0.05) observed for area under the curve responses of insulin, glucose, nonesterified fatty acids, or betahydroxybutryrate. SIC reduces postprandial lipemia, but the energy deficit alone does not fully explain the decrease observed.
Arm exercise stress perfusion imaging predicts clinical outcome
Authors: Chan Albert K. , Ilias-Khan Nasreen A. , Xian Hong , Inman Cindi , Martin Wade H.
Published: August 2011

Treadmill exercise capacity in resting metabolic equivalents (METs) and stress hemodynamic, electrocardiographic (ECG), and myocardial perfusion imaging (MPI) responses are independently predictive of adverse clinical events. However, limited data exist for arm ergometer stress testing (AXT) in patients who cannot perform leg exercise because of lower extremity disabilities. We sought to determine the extent to which AXT METs, hemodynamic, ECG, and MPI responses to arm exercise add independent incremental value to demographic and clinical variables for prediction of all-cause mortality, myocardial infarction (MI), or late coronary revascularization, individually or as a composite. A prospective cohort of 186 patients aged 64 ± 10 (SD) yr, unable to perform lower extremity exercise, underwent AXT MPI for clinical reasons between 1997 and 2002, and were followed for 62 ± 23 mo, to an endpoint of death or 12/31/2006. Average annual rates were 5.4% for mortality, 2.2% for MI, 2.5% for late coronary revascularization, and 8.0% for combined events. After adjustment for age and clinical variables, AXT METs [ P < 0.05; hazard ratio (HR) = 0.59; confidence interval (CI) = 0.35–0.84] and abnormal MPI ( P < 0.01; HR = 2.48; CI = 2.15–2.81) were independently predictive of mortality. A positive AXT ECG ( P < 0.05; HR = 2.61; CI = 2.13–3.10) was predictive of MI. Death and MI combined were prognosticated by METs ( P < 0.05; HR = 0.63; CI = 0.41–0.85), MPI ( P < 0.05; HR = 1.77; CI = 1.49–2.05), and a positive AXT ECG ( P < 0.05; HR = 1.86; CI = 1.55–2.17). In conclusion, for high risk older patients who cannot perform leg exercise because of lower extremity disabilities, AXT METs are as important as MPI for prediction of mortality alone and death and MI combined, and a positive AXT ECG prognosticates MI alone and death and MI combined.
Takotsubo Cardiomyopathy “Variations on a Theme”
Authors: Bogaard Kjell , Odekerken Diego A. M. , Voogel Albertus J.
Published: August 2011

Takotsubo Cardiomyopathy (TTC) is a fairly new diagnosis in the cardiologist's repertoire. It can present itself in multiple different forms. We describe three cases of TTC with different etiologies illustrating the broad spectrum of presentations.
Congenital Intranasal Glioma
Authors: Salati Sajad Ahmad , Rather Ajaz Ahmad
Published: August 2011

Congenital midline swellings of nose are encountered rarely, and nasal gliomas constitute about 5% of such lesions. Various theories have been suggested to explain the pathogenesis. Imaging preferably by MRI is mandated to study the extent and to rule out intracranial extension. Treatment is complete excision, and the approach depends upon the extent of the lesion and availability of expertise. We present the management of one such case of congenital intranasal glioma without any intracranial extension that presented as a septal polyp.
A Case of Severe Asymptomatic Aortic Coarctation in an Adult Female
Authors: Tsikrikas Spyros , Charalampous Charalampos , Letsas Konstantinos P.
Published: August 2011

Coarctation of the aorta is typically a disease of childhood and early adulthood, and there is a reduced life expectancy in patients who have not undergone correction. Survival to older age is rare, due to severe cardiovascular complications. We describe the case of a woman first diagnosed with coarctation of aorta at an advanced age.
MicroRNA-708 Induces Apoptosis and Suppresses Tumorigenicity in Renal Cancer Cells
Authors: Saini Sharanjot , Yamamura Soichiro , Majid Shahana , Shahryari Varahram , Hirata Hiroshi , Tanaka Yuichiro , Dahiya Rajvir
Published: August 2011

Cancer pathogenesis is restricted by stresses that compromise cell division and survival. In this study, we identify miR-708, a little studied member of a set of microRNAs that have been implicated in stress control, as an important tumor suppressor in renal cell carcinoma (RCC). miR-708 expression was attenuated widely in human RCC specimens. Restoration of miR-708 expression in RCC cell lines decreased cell growth, clonability, invasion, and migration and elicited a dramatic increase in apoptosis. Moreover, intratumoral delivery of miR-708 was sufficient to trigger in vivo regression of established tumors in murine xenograft models of human RCC. Investigation of the targets of miR-708 identified the inhibitor of apoptosis protein survivin as important. siRNA-mediated knockdown of survivin partially phenocopied miR-708 overexpression suggesting that the proapoptotic role of miR-708 may be mediated primarily through survivin regulation. Additionally, we identified the E-cadherin regulators ZEB2 and BMI1 as likely miR-708 targets. Taken together, our findings define a major tumor suppressive role for miR-708, which may offer an attractive new target for prognostic and therapeutic intervention in RCC. Cancer Res; 71(19); 6208–19. ©2011 AACR.
Haploinsufficiency in the Prometastasis Kiss1 Receptor Gpr54 Delays Breast Tumor Initiation, Progression, and Lung Metastasis
Authors: Cho Sung-Gook , Wang Ying , Rodriguez Melissa , Tan Kunrong , Zhang Wenzheng , Luo Jian , Li Dali , Liu Mingyao
Published: August 2011

Activation of KISS1 receptor (KISS1R or GPR54) by its ligands (Kisspeptins) regulates a diverse function both in normal physiology and pathophysiology. In cancer, KISS1R has been implicated in tumor angiogenesis and metastasis, but a broader evaluation of KISS1R in tumorigenesis and tumor progression is yet to be conducted. In this study, we used mouse models of Kiss1r gene knockout and mouse mammary tumor virus–polyoma virus middle T antigen (MMTV-PyMT)–induced breast cancer to conduct such an evaluation. Kiss1r heterozygosity in MMTV-PyMT mice was sufficient to attenuate breast cancer initiation, growth, latency, multiplicity, and lung metastasis. To confirm these effects and assess possible contributions of endogenous ligands, we isolated primary tumor cells from PyMT/Kiss1r+/+ and PyMT/Kiss1r+/− mice and compared their phenotypes by in vitro and in vivo assays. Kiss1r loss attenuated in vitro tumorigenic properties as well as tumor growth in vivo in immunocompromised NOD.SCID/NCr mice. Kiss1r activation in these cells, resulting from the addition of its ligand Kisspeptin-10, resulted in RhoA activation and RhoA-dependent gene expression through the Gαq-p63RhoGEF signaling pathway. Anchorage-independent growth was tightly linked to dose-dependent regulation of RhoA by Kiss1r. In support of these results, siRNA-mediated knockdown of KISS1R or inactivation of RhoA in human MCF10A breast epithelial cells overexpressing H-RasV12 was sufficient to reduce Ras-induced anchorage-independent growth. In summary, we concluded that Kiss1r attenuation was sufficient to delay breast tumor initiation, progression, and metastasis through inhibitory effects on the downstream Gαq-p63RhoGEF-RhoA signaling pathway. Cancer Res; 71(20); 6535–46. ©2011 AACR.
Early Detection of Tumor Cells by Innate Immune Cells Leads to Treg Recruitment through CCL22 Production by Tumor Cells
Authors: Faget Julien , Biota Cathy , Bachelot Thomas , Gobert Michael , Treilleux Isabelle , Goutagny Nadège , Durand Isabelle , Léon-Goddard Sophie , Blay Jean Yves , Caux Christophe , Ménétrier-Caux Christine
Published: August 2011

In breast carcinomas, patient survival seems to be negatively affected by the recruitment of regulatory T cells (Treg) within lymphoid aggregates by CCL22. However, the mechanisms underpinning this process, which may be of broader significance in solid tumors, have yet to be described. In this study, we determined how CCL22 production is controlled in tumor cells. In human breast carcinoma cell lines, CCL22 was secreted at low basal levels that were strongly increased in response to inflammatory signals [TNF-α, IFN-γ, and interleukin (IL)-1β], contrasting with CCL17. Primary breast tumors and CD45+ infiltrating immune cells appeared to cooperate in driving CCL22 secretion, as shown clearly in cocultures of breast tumor cell lines and peripheral blood mononuclear cells (PBMC) or their supernatants. We determined that monocyte-derived IL-1β and TNF-α are key players as monocyte depletion or neutralization of these cytokines attenuated secretion of CCL22. However, when purified monocytes were used, exogenous human IFN-γ was also required to generate this response suggesting a role for IFN-γ–producing cells within PBMCs. In this setting, we found that human IFN-γ could be replaced by the addition of (i) IL-2 or K562-activated natural killer (NK) cells or (ii) resting NK cells in the presence of anti-MHC class I antibody. Taken together, our results show a dialogue between NK and tumor cells leading to IFN-γ secretion, which in turn associates with monocyte-derived IL-1β and TNF-α to drive production of CCL22 by tumor cells and subsequent recruitment of Treg. As one validation of this conclusion in primary breast tumors, we showed that NK cells and macrophages tend to colocalize within tumors. In summary, our findings suggest that at early times during tumorigenesis, the detection of tumor cells by innate effectors (monocytes and NK cells) imposes a selection for CCL22 secretion that recruits Treg to evade this early antitumor immune response. Cancer Res; 71(19); 6143–52. ©2011 AACR.
Oncogene AEG-1 Promotes Glioma-Induced Neurodegeneration by Increasing Glutamate Excitotoxicity
Authors: Lee Seok-Geun , Kim Keetae , Kegelman Timothy P. , Dash Rupesh , Das Swadesh K. , Choi Jung Kyoung , Emdad Luni , Howlett Eric L. , Jeon Hyun Yong , Su Zhao Zhong , Yoo Byoung Kwon , Sarkar Devanand , Kim Sung-Hoon , Kang Dong-Chul , Fisher Paul B.
Published: August 2011

Aggressive tumor growth, diffuse tissue invasion, and neurodegeneration are hallmarks of malignant glioma. Although glutamate excitotoxicity is considered to play a key role in glioma-induced neurodegeneration, the mechanism(s) controlling this process is poorly understood. Astrocyte elevated gene-1 (AEG-1) is an oncogene that is overexpressed in several types of human cancers, including more than 90% of brain tumors. In addition, AEG-1 promotes gliomagenesis, particularly in the context of tumor growth and invasion, 2 primary characteristics of glioma. In the present study, we investigated the contribution of AEG-1 to glioma-induced neurodegeneration. Pearson correlation coefficient analysis in normal brain tissues and samples from glioma patients indicated a strong negative correlation between expression of AEG-1 and a primary glutamate transporter of astrocytes EAAT2. Gain- and loss-of-function studies in normal primary human fetal astrocytes and T98G glioblastoma multiforme cells revealed that AEG-1 repressed EAAT2 expression at a transcriptional level by inducing YY1 activity to inhibit CBP function as a coactivator on the EAAT2 promoter. In addition, AEG-1–mediated EAAT2 repression caused a reduction of glutamate uptake by glial cells, resulting in induction of neuronal cell death. These findings were also confirmed in samples from glioma patients showing that AEG-1 expression negatively correlated with NeuN expression. Taken together, our findings suggest that AEG-1 contributes to glioma-induced neurodegeneration, a hallmark of this fatal tumor, through regulation of EAAT2 expression. Cancer Res; 71(20); 6514–23. ©2011 AACR.
ZNF668 Functions as a Tumor Suppressor by Regulating p53 Stability and Function in Breast Cancer
Authors: Hu Ruozhen , Peng Guang , Dai Hui , Breuer Eun-Kyoung , Stemke-Hale Katherine , Li Kaiyi , Gonzalez-Angulo Ana M. , Mills Gordon B. , Lin Shiaw-Yih
Published: August 2011

Genome-wide sequencing studies in breast cancer have recently identified frequent mutations in the zinc finger protein 668 (ZNF668), the function of which is undefined. Here, we report that ZNF668 is a nucleolar protein that physically interacts with and regulates p53 and its negative regulator MDM2. Through MDM2 binding, ZNF668 regulated autoubiquitination of MDM2 and its ability to mediate p53 ubiquitination and degradation. ZNF668 deficiency also impaired DNA damage–induced stabilization of p53. RNA interference–mediated knockdown of ZNF668 was sufficient to transform normal mammary epithelial cells. ZNF668 effectively suppressed breast cancer cell proliferation in vitro and tumorigenicity in vivo. Taken together, our studies identify ZNF668 as a novel breast tumor suppressor gene that functions in regulating p53 stability. Cancer Res; 71(20); 6524–34. ©2011 AACR.
A Mutation Threshold Distinguishes the Antitumorigenic Effects of the Mitochondrial Gene MTND1, an Oncojanus Function
Authors: Gasparre Giuseppe , Kurelac Ivana , Capristo Mariantonietta , Iommarini Luisa , Ghelli Anna , Ceccarelli Claudio , Nicoletti Giordano , Nanni Patrizia , De Giovanni Carla , Scotlandi Katia , Betts Christine M. , Carelli Valerio , Lollini Pier Luigi , Romeo Giovanni , Rugolo Michela , Porcelli Anna Maria
Published: August 2011

The oncogenic versus suppressor roles of mitochondrial genes have long been debated. Peculiar features of mitochondrial genetics such as hetero/homoplasmy and mutation threshold are seldom taken into account in this debate. Mitochondrial DNA (mtDNA) mutations generally have been claimed to be protumorigenic, but they are also hallmarks of mostly benign oncocytic tumors wherein they help reduce adaptation to hypoxia by destabilizing hypoxia-inducible factor-1α (HIF1α). To determine the influence of a disassembling mtDNA mutation and its hetero/homoplasmy on tumorigenic and metastatic potential, we injected mice with tumor cells harboring different loads of the gene MTND1 m.3571insC. Cell cultures obtained from tumor xenografts were then analyzed to correlate energetic competence, apoptosis, α-ketoglutarate (α-KG)/succinate (SA) ratio, and HIF1α stabilization with the mutation load. A threshold level for the antitumorigenic effect of MTND1 m.3571insC mutation was defined, above which tumor growth and invasiveness were reduced significantly. Notably, HIF1α destabilization and downregulation of HIF1α-dependent genes occurred in cells and tumors lacking complex I (CI), where there was an associated imbalance of α-KG/SA despite the presence of an actual hypoxic environment. These results strongly implicate mtDNA mutations as a cause of oncocytic transformation. Thus, the antitumorigenic and antimetastatic effects of high loads of MTND1 m.3571insC, following CI disassembly, define a novel threshold-regulated class of cancer genes. We suggest these genes be termed oncojanus genes to recognize their ability to contribute either oncogenic or suppressive functions in mitochondrial settings during tumorigenesis. Cancer Res; 71(19); 6220–9. ©2011 AACR.
High-Dose Antiangiogenic Therapy for Glioblastoma: Less May Be More?
Authors: de Groot John F.
Published: August 2011

Targeting angiogenesis in glioblastoma rapidly reduces vascular permeability and contrast enhancement on MRI and prolongs progression-free survival. The long-term efficacy of bevacizumab and other antiangiogenic agents is limited, however, because of the rapid development of resistance. Alternative dosing approaches may be one mechanism of prolonging therapeutic efficacy. Clin Cancer Res; 17(19); 6109–11. ©2011 AACR.
Importance of Phagosomal Functionality for Growth Restriction of Mycobacterium tuberculosis in Primary Human Macrophages
Authors: Welin Amanda , Raffetseder Johanna , Eklund Daniel , Stendahl Olle , Lerm Maria
Published: August 2011

The best characterized survival mechanism of Mycobacterium tuberculosis inside the macrophage is the inhibition of phagosomal maturation. Phagosomal maturation involves several steps including fusion with lysosomes and acidification. However, it has not been elucidated which components of phagosomal maturation correlate with growth restriction of virulent mycobacteria in human macrophages, and we aimed to study this. We infected human monocyte-derived macrophages with M. tuberculosis and assessed bacterial replication, translocation of CD63 to the phagosome, and phagosomal acidification. We found that unstimulated human macrophages were able to control infection with M. tuberculosis upon inoculation at a low multiplicity of infection (MOI) of 1, but not at a high MOI of 10. The low MOI resulted in a macrophage-controlled balance between host cells and bacteria. Both H37Rv and H37Ra infection, at high and low MOI, led to equally ineffective translocation of CD63 to the phagosome. On the other hand, acidification of mycobacterial phagosomes was more efficient at MOI 1 than 10 with both mycobacterial strains, consistent with a direct or indirect role for phagosomal acidification in restricting M. tuberculosis growth. Furthermore, inhibition of the vacuolar H+-ATPase as well as of cathepsin D led to enhanced mycobacterial replication inside the macrophage. This again shows the importance of phagosomal acidification for control of mycobacterial growth, through the activation of lysosomal hydrolases. We conclude that acidification and related functional aspects of the mature phagosome are important factors for restriction of M. tuberculosis replication in human macrophages.
Failure of a Vagus Nerve Stimulator following a Nearby Lightning Strike
Authors: Terry Garth E. , Conry Joan A. , Taranto Eleanor , Yaun Amanda
Published: August 2011

We recently reported our experience with implanted vagus nerve stimulators (VNS) in 62 children over a 7-year period. Here, we present a case of a VNS that successfully reduced the number and severity of seizures in a patient with an unusual seizure pattern, and failed to function shortly after a lightning storm. To our knowledge, the failure of VNS or any implantable electrical devices by lightning has not been reported in the literature. This mechanism of electrical interference, while unusual, may require more attention as these devices are expected to be used more frequently.
Brain Transcriptomic Response of Threespine Sticklebacks to Cues of a Predator
Authors: Sanogo Yibayiri O. , Hankison Shala , Band Mark , Obregon Alexandra , Bell Alison M.
Published: August 2011

Predation pressure represents a strong selective force that influences the development and evolution of living organisms. An increasing number of studies have shown that both environmental and social factors, including exposure to predators, substantially shape the structure and function of the brain. However, our knowledge about the molecular mechanisms underlying the response of the brain to environmental stimuli is limited. In this study, we used whole-genome comparative oligonucleotide microarrays to investigate the brain transcriptomic response to cues of a predator in the threespine stickleback, Gasterosteus aculeatus. We found that repeated exposure to olfactory, visual and tactile cues of a predator (rainbow trout, Oncorrhynchus mykiss) for 6 days resulted in subtle but significant transcriptomic changes in the brain of sticklebacks. Gene functional analysis and gene ontology enrichment revealed that the majority of the transcripts differentially expressed between the fish exposed to cues of a predator and the control group were related to antigen processing and presentation involving the major histocompatibility complex, transmission of synaptic signals, brain metabolic processes, gene regulation and visual perception. The top four identified pathways were synaptic long-term depression, RAN signaling, relaxin signaling and phototransduction. Our study demonstrates that exposure of sticklebacks to cues of a predator results in the activation of a wide range of biological and molecular processes and lays the foundation for future investigations on the molecular factors that modulate the function and evolution of the brain in response to stressors.
Are Wet-Induced Wrinkled Fingers Primate Rain Treads?
Authors: Changizi Mark , Weber Romann , Kotecha Ritesh , Palazzo Joseph
Published: August 2011

Wet fingers and toes eventually wrinkle, and this is commonly attributed by lay opinion to local osmotic reactions. However, nearly a century ago surgeons observed that no wrinkling occurs if a nerve to the finger has been cut. Here we provide evidence that, rather than being an accidental side effect of wetness, wet-induced wrinkles have been selected to enhance grip in wet conditions. We show that their morphology has the signature properties of drainage networks, enabling efficient removal of water from the gripped surface.
The Effect of HMGB1, a Damage-Associated Molecular Pattern Molecule, on Polymorphonuclear Neutrophil Migration Depends on Its Concentration
Authors: Berthelot Florence , Fattoum Lakhdar , Casulli Sarah , Gozlan Joël , Maréchal Vincent , Elbim Carole
Published: August 2011

Polymorphonuclear neutrophils (PMN) play a key role in host defenses against invading microorganisms but also potentiate inflammatory reactions in case of excessive or misdirected responses. Release of the alarmin high-mobility group box 1 (HMGB1) by cells that die at an inflammatory site may act as an alert signal for the immune system. We studied the effect of HMGB1 on human PMN migration, using whole-blood samples to avoid cell activation associated with isolation procedures. HMGB1 50–100 ng/ml reduced baseline PMN migration as well as formyl-methionyl-leucyl-phenylalanine- and IL-8-induced PMN chemotaxis. This inhibitory effect was mediated by the RAGE receptor. In contrast, a higher HMGB1 concentration (5,000 ng/ml) had a chemoattractant effect on PMN through IL-8 production. This effect required the engagement of Toll-like receptors 2 and 4 in addition to the RAGE receptor. The A box component of HMGB1, which antagonizes the endogenous protein, reduced chemotaxis and also strongly inhibited the enhancement of PMN migration observed with the highest HMGB1 concentration. In contrast, the B box, reported to be the active form of HMGB1, exerted a chemoattractant effect. These results strongly point to a key regulatory role of HMGB1 in PMN recruitment to inflammatory tissues. The A box component could potentially serve to inhibit inappropriate PMN recruitment during chronic inflammatory disorders associated with excessive HMGB1 release.
Localization of Calbindin-D28k and Calretinin in the Brain of Dermophis Mexicanus (Amphibia: Gymnophiona) and Its Bearing on the Interpretation of Newly Recognized Neuroanatomical Regions
Authors: Morona Ruth , López Jesús María , González Agustín
Published: August 2011

The analysis of the distribution of the calbindin-D28k and calretinin immunoreactive (CBir and CRir) systems recently described in the brain of anuran and urodele amphibians was very useful for the interpretation of many otherwise indistinct brain regions and cell masses. In the present study we have followed a similar approach to investigate the distribution of CBir and CRir cell bodies and fibers in the brain of Dermophis mexicanus, a member of the much neglected third amphibian order of gymnophionans. The pattern of distribution obtained showed particular characteristics in Dermophis, such as the existence of abundant CRir elements in the olfactory bulbs and CBir and CRir cell populations in pallial areas. The distinct distribution of the two proteins allowed the tentative identification of currently described subregions, mainly in the amygdaloid complex and hypothalamic areas. The analysis of the diencephalon and brainstem distribution framed in the neuromeric model highlighted common traits with other amphibians but also specific features. Therefore, the immunohistochemical detection of calcium-binding proteins has served to discern cell populations and has helped to demonstrate neuronal heterogeneity. However, it should be pointed out that a straightforward comparison based only on the presence of these proteins should not be made due to the great variability observed in well-established homologous regions in the brain of different vertebrates, as evidenced within the class Amphibia.
Candida albicans-Induced Chronic Thrombocytopenic Purpura
Authors: Ayesh (Haj Yousef) Mahmoud H. , Alawneh Khaldoon M.
Published: August 2011

We present 2 patients with chronic immune thrombocytopenic purpura (ITP) secondary to Candida albicans infection. Neither patient responded to standard ITP therapy including splenectomy. Appropriate antifungal treatment of the C. albicans infection was followed by sustained improvement in platelet count in both patients. To our knowledge, this is the first report of ITP in association with C. albicans infection.
Activity of Matrix Metalloproteinases in Bovine versus Human Dentine
Authors: Kato M.T. , Hannas A.R. , Leite A.L. , Bolanho A. , Zarella B.L. , Santos J. , Carrilho M. , Tjäderhane L. , Buzalaf M.A.R.
Published: August 2011

Metalloproteinases (MMPs) have been implicated with metabolism of collagen in physiological and pathological processes in human dentine. As bovine teeth have been used as a substitute for human teeth in laboratory analysis, this study evaluated the activity of MMP-2 and -9 in bovine versus human dentine. Bovine and human dentine fragments, from crowns and roots, were powderized. Protein extraction was performed by two protocols: a neutral extraction with guanidine-HCl/EDTA (pH 7.4) and an acidic extraction with citric acid (pH 2.3). Gelatinolytic activities of extracts were revealed by zymography. MMP-2 and -9 were detected in crown and root dentine from bovine and human teeth. Total activities of MMP-2 were 11.4 ± 2.2, 14.6 ± 2.0, 9.7 ± 1.2 and 12.4 ± 0.9 ng/ml for bovine root, human root, bovine crown and human crown dentine, respectively. Corresponding activities for MMP-9 were 14.9 ± 2.0, 15.3 ± 1.3, 15.4 ± 1.3 and 15.5 ± 1.3 ng/ml, respectively. Bovine dentine was found to be a reliable substrate for studies involving the activity of MMP-2 and -9.
Thrombospondin-1 Short Hairpin RNA Suppresses Tubulointerstitial Fibrosis in the Kidney of Ureteral Obstruction by Ameliorating Peritubular Capillary Injury
Authors: Sun Dong , Ma Yanyan , Han Hong , Yin Zhongcheng , Liu Caixia , Feng Jinhong , Zhou Xudong , Li Xiaoju , Xiao Aiguo , Yu Rong
Published: August 2011

Background/Aims: Thrombospondin-1 (TSP-1), a naturally occurring inhibitor of angiogenesis, is an important mediator of renal fibrosis in clinical and experimental kidney disease. Increasing evidence shows that the microvasculature plays a critical role in progressive renal disease. The present study was undertaken to investigate whether interstitial fibrosis could be prevented by abolishing TSP-1 function in unilateral ureteral obstruction (UUO)-induced renal fibrosis. Methods and Results: A short hairpin RNA vector, designated Thbs-1, significantly suppressed TSP-1 in both transcriptional and translational levels in in vitro-cultured cells and in vivo fibrosis-induced mouse kidney. Furthermore, TSP-1 RNA interference increased the protein level of vascular endothelial growth factor (VEGF) and the density of peritubular capillaries (PTCs), reduced the expression of hypoxia-inducible factor-1α in tubulointerstitial cells, and collagen III and the connective tissue growth factor expression were markedly reduced from day 7 after UUO-induced fibrosis, but un- or vector-treated mice maintained their expression. TSP-1 shRNA suppressed the protein level of TSP-1, increased VEGF expression and PTC density and alleviated the development of renal interstitial fibrosis in UUO mice. Conclusion: These data suggest that inhibition of TSP-1 expression prevented tubulointerstitial fibrosis through ameliorating PTC injury.
Comparison of the Proliferation, Viability, and Differentiation Capacity of Adipose-Derived Stem Cells from Different Anatomic Sites in Rabbits
Authors: Chen L. , Peng E.J. , Zeng X.Y. , Zhuang Q.Y. , Ye Z.Q.
Published: August 2011

Tissue engineering is clinically promising for missing and damaged tissues. Adipose-derived stem cells (ASCs), a type of mesenchymal stem cells, represent a reliable source of seed cells for tissue engineering with multiple merits such as minimal invasion, abundant yield, little immunity, low morbidity, easy isolation, and rapid expansion. However, because the properties of adipose tissue-derived cells differ depending on the fat depot from which they are derived, we compared the ASCs from three anatomic sites of New Zealand white rabbits: subcutaneous inguinal (SI), subcutaneous dorsocervical (SD), and retroperitoneal perinephric (RP) regions. We investigated cellular behaviors including proliferation, viability, and differentiation. The ASCs of the subcutaneous regions (SI and SD) had higher performances in all assessments compared to those of the RP region. Moreover, the SI and SD ASCs had significant differences, with SI ASCs having better properties than SD ASCs. We conclude that the different anatomic distributions of fat contribute to the different behaviors of ASCs. The SI region offers the most applicable cell source reservoir for ASC tissue engineering.
AMPK Alpha 1-Induced RhoA Phosphorylation Mediates Vasoprotective Effect of Estradiol
Authors: Gayard Marion , Guilluy Christophe , Rousselle Anthony , Viollet Benoit , Henrion Daniel , Pacaud Pierre , Loirand Gervaise , Rolli-Derkinderen Malvyne
Published: August 2011

Objective—

Estradiol (E2) mediates numerous beneficial effects assigned to estrogens, but whereas mechanisms have been described at the endothelial level, direct effects on vascular smooth muscle cells (VSMC) are poorly documented. As evidence accumulates regarding the role of RhoA in vascular pathophysiology and the benefit of RhoA-Rho associated protein kinase (Rock) pathway inhibition, we analyzed if E2 could inhibit it in VSMC.

Methods and Results—

We show that in VSMC, E2 inhibits the RhoA-Rock pathway in a time- and concentration-dependent manner. The inhibition of RhoA-Rock pathway results from E2-induced phosphorylation of the Ser188 of RhoA. Using pharmacological, transfection, and in vitro phosphorylation experiments, we demonstrate that AMP-activated protein kinase subunit alpha 1 (AMPKα1) is activated by estrogen receptor stimulation and catalyzes RhoA phosphorylation induced by E2. Ex vivo, ovariectomy leads to an increase in the amplitude of phenylephrine- or serotonine-induced contractions of aortic rings in wild-type mice but not in AMPKα1-knock-out mice or E2-supplemented animals. These functional effects were correlated with a reduced level of RhoA phosphorylation in the aorta of ovariectomized female, male, and AMPKα1 knock-out mice.

Conclusion—

Our work thus defines AMPKα1 as (1) a new kinase for RhoA and (2) a new mediator of the vasoprotective effects of estrogen.
Nitric Oxide Decreases the Expression of Endothelin-Converting Enzyme-1 Through mRNA Destabilization
Authors: Raoch Viviana , Rodríguez-Pascual Fernando , López-Martínez Vanesa , Medrano-Andrés Diana , Rodríguez-Puyol Manuel , Lamas Santiago , Rodríguez-Puyol Diego , López-Ongil Susana
Published: August 2011

Objective—

Endothelial function depends on the equilibrium in the synthesis of vasoactive endothelial factors. It is well known that endothelin and nitric oxide (NO) exhibit reciprocal regulation. We assessed the ability of NO to regulate endothelin-converting enzyme-1 (ECE-1) expression in vascular endothelial cells.

Methods and Results—

Bovine aortic endothelial cells were incubated with 2 different NO donors as well as with a cyclic-GMP analog, dibutyryl-cGMP (dB-cGMP). ECE-1 protein content and mRNA expression were evaluated by Western blot and Northern blot, respectively, promoter activity by transfection experiments, ECE-1 activity by ELISA, and cGMP production by radioimmunoassay. Both NO donors decreased ECE-1 protein content, mRNA expression, and ECE-1 activity. ODQ, an inhibitor of soluble guanylyl cyclase, blocked those effects. NO donors raised cGMP levels, and dB-cGMP mimicked their effects on ECE-1 expression, which were blocked by KT5823, a nonspecific PKG inhibitor. The changes on ECE-1 expression were due to a destabilization on 3′-untranslated region (3′-UTR) of this mRNA, because the activity of a luciferase reporter construct containing the 3′-UTR of the ECE-1 gene was reduced by dB-cGMP in a PKG-dependent manner. The biological relevance of this regulation was confirmed in bovine aortic endothelial cells coincubated with macrophages in the presence of lipopolysaccharide, in eNOS-deficient mice, and in Wistar rats treated with NO donors. In every case, an inverse relationship was observed between NO and ECE-1 protein content.

Conclusion—

Our results support that NO regulates ECE-1 expression through a cGMP/PKG-dependent regulatory mechanism at the post-transcriptional level via the 3′-UTR of the ECE-1 gene.
Focal Adhesion Kinase–Related Nonkinase Inhibits Vascular Smooth Muscle Cell Invasion by Focal Adhesion Targeting, Tyrosine 168 Phosphorylation, and Competition for p130
Cas
Binding
Authors: Koshman Yevgeniya E. , Chu Miensheng , Engman Steven J. , Kim Taehoon , Iyengar Rekha , Robia Seth L. , Samarel Allen M.
Published: August 2011

Objective—

Focal adhesion kinase–related nonkinase (FRNK), the C-terminal domain of focal adhesion kinase (FAK), is a tyrosine-phosphorylated, vascular smooth muscle cell (VSMC)–specific inhibitor of cell migration. FRNK inhibits both FAK and proline-rich tyrosine kinase 2 (PYK2) in cultured VSMCs, and both kinases may be involved in VSMC invasion during vascular remodeling.

Methods and Results—

Adenovirally mediated gene transfer of green fluorescent protein–tagged, wild-type (wt) FRNK into balloon-injured rat carotid arteries confirmed that FRNK overexpression inhibited both FAK and PYK2 phosphorylation and downstream signaling in vivo. To identify which kinase was involved in regulating VSMC invasion, adenovirally mediated expression of specific short hairpin RNAs was used to knock down FAK versus PYK2 in cultured VSMCs, but only FAK short hairpin RNA was effective in reducing VSMC invasion. The role of FRNK tyrosine phosphorylation was then examined using adenoviruses expressing nonphosphorylatable (Tyr168Phe-, Tyr232Phe-, and Tyr168,232Phe-) green fluorescent protein–FRNK mutants. wtFRNK and all FRNK mutants localized to FAs, but only Tyr168 phosphorylation was required for FRNK to inhibit invasion. Preventing Tyr168 phosphorylation also increased FRNK-paxillin interaction, as determined by coimmunoprecipitation, total internal reflection fluorescence microscopy, and fluorescence recovery after photobleaching. Furthermore, wtFRNK competed with FAK for binding to p130
Cas
(a critically important regulator of cell migration) and prevented its phosphorylation. However, Tyr168Phe-FRNK was unable to bind p130
Cas
.

Conclusion—

We propose a 3-stage mechanism for FRNK inhibition: focal adhesion targeting, Tyr168 phosphorylation, and competition with FAK for p130
Cas
binding and phosphorylation, which are all required for FRNK to inhibit VSMC invasion.
The Coronary Artery Disease–Associated 9p21 Variant and Later Life 20-Year Survival to Cohort Extinction
Authors: Dutta Ambarish , Henley William , Lang Iain A. , Murray Anna , Guralnik Jack , Wallace Robert B. , Melzer David
Published: August 2011

Background—

Common variation at chromosome 9p21 (marked by
rs10757278
or
rs1333049
) is associated with coronary artery disease (CAD) and peripheral vascular disease. A decreasing effect at older age was suggested, and effects on long-term mortality are unclear. We estimated 9p21 associations with CAD and all-cause mortality in a CAD diagnosis–free older population. We also estimated classification gains on adding the variant to the Framingham Risk Score (FRS) for CAD.

Methods and Results—

DNA was from an Established Populations for Epidemiological Study of the Elderly–Iowa cohort from 1988 (participants >71 years), with death certificates obtained to 2008 for 92% of participants. Cox regression models were adjusted for confounders and CAD risk factors. Of 1095 CAD diagnosis–free participants, 52% were heterozygous (CG) and 22% were homozygous (CC) for the risk
C
allele
rs1333049
. Unadjusted CAD-attributed death rates in the CC group were 30 vs 22 per 1000 person-years for the GG group. The
C
allele was associated with all-cause (hazard ratio, 1.19; 95% CI, 1.08–1.30) and CAD (hazard ratio, 1.29; 95% CI, 1.08–1.56) mortality, independent of CAD risk factors. There was no association with stroke deaths. Variant associations with CAD mortality were attenuated after the age of 80 years (age-interaction term
P
=0.05). In age group 71 to 80 years, FRS classified as high risk 21% of respondents who died of CAD within 10 years; adding 9p21 identified 27% of respondents.

Conclusions—

In 71- to 80-year-old subjects free of CAD diagnoses, 9p21 is associated with excess mortality, mainly attributed to CAD mortality. Adding 9p21 to the FRS may improve the targeting of CAD prevention in older people, but validation in independent samples is needed for confirmation.
Carbon Monoxide–Activated Nrf2 Pathway Leads to Protection Against Permanent Focal Cerebral Ischemia
Authors: Wang Bing , Cao Wangsen , Biswal Shyam , Doré Sylvain
Published: August 2011

Background and Purpose—

Carbon monoxide (CO) is a gaseous second messenger produced when heme oxygenase enzymes catabolize heme. We have demonstrated that CO can be therapeutic in ischemia-reperfusion brain injury; however, it is unclear whether CO can also offer protection in permanent ischemic stroke or what mechanism(s) underlies the effect. Heme oxygenase-1 neuroprotection was shown to be regulated by Nrf2; therefore, we investigated whether CO might partially exert neuroprotection by modulating the Nrf2 pathway.

Methods—

To evaluate the potential protective effects of CO, we exposed male wild-type and Nrf2-knockout mice to 250 ppm CO or control air for 18 hours immediately after permanent middle cerebral artery occlusion. Infarct volume and neurologic deficits were assessed on day 7. Molecular mechanisms of Nrf2 pathway activation by CO were also investigated.

Results—

Mice exposed to CO after permanent ischemia had 29.6±12.6% less brain damage than did controls at 7 days, although amelioration in neurologic deficits did not reach significance. Additionally, 18-hour CO treatment led to Nrf2 dissociation from Keap1, nuclear translocation, increased binding activity of Nrf2 to heme oxygenase-1 antioxidant response elements, and elevated heme oxygenase-1 expression 6 to 48 hours after CO exposure. The CO neuroprotection was completely abolished in Nrf2-knockout mice.

Conclusions—

Low-concentration CO represent a neuroprotective agent for combination treatment of ischemic stroke, and its beneficial effect would be at least partially mediated by activation of the Nrf2 pathway.
Microembolism Versus Hemodynamic Impairment in Rosary-Like Deep Watershed Infarcts
Authors: Moustafa Ramez Reda , Momjian-Mayor Isabelle , Jones P. Simon , Morbelli Silvia , Day Diana J. , Aigbirhio Franklin I. , Fryer Tim D. , Warburton Elizabeth A. , Baron Jean-Claude
Published: August 2011

Background and Purpose—

Deep watershed infarcts are frequent in high-grade carotid disease and are thought to result from hemodynamic impairment, particularly when adopting a rosary-like pattern. However, a role for microembolism has also been suggested, though never directly tested. Here, we studied the relationships among microembolic signals (MES) on transcranial Doppler, rosary-like deep watershed infarcts on brain imaging, and cerebral hemodynamic compromise on positron emission tomography (PET), all in severe symptomatic carotid disease. We hypothesized that rosary-like infarcts would be significantly associated with worse hemodynamic status, independent of the presence of MES.

Methods—

Sixteen patients with ≥70% carotid disease ipsilateral to recent transient ischemic attack/minor stroke underwent magnetic resonance imaging including diffusion-weighted imaging,
15
O-PET, and transcranial Doppler. Mean transit time, a specific marker for hemodynamic impairment, was obtained in the symptomatic and unaffected hemispheres.

Results—

Eleven of 16 patients had rosary-like infarcts (Rosary+) and 8 patients had MES. Mean transit time was significantly higher (
P
=0.008) in Rosary+ patients than in healthy controls (
n
=10), and prevalence of MES was not different between Rosary+ and Rosary− patients. Contrary to our hypothesis, however, the presence of MES within the Rosary+ subset was associated (
P
=0.03) with a better hemodynamic status than in their absence, with a significant (
P
=0.02) negative correlation between mean transit time and rate of MES/h.

Conclusions—

Contrary to mainstream understanding, rosary-like infarcts were not independent of presence and rate of MES, suggesting that microembolism plays a role in their pathogenesis, probably in association with hemodynamic impairment. Pending confirmation in a larger sample, these findings have management implications for patients with carotid disease and rosary-like infarcts.
Impact of the Extended Thrombolysis Time Window on the Proportion of Recombinant Tissue-Type Plasminogen Activator-Treated Stroke Patients and on Door-to-Needle Time
Authors: Minnerup Jens , Wersching Heike , Ringelstein E. Bernd , Schilling Matthias , Schäbitz Wolf-Rüdiger , Wellmann Jürgen , Berger Klaus
Published: August 2011

Background and Purpose—

The European Cooperative Acute Stroke Study (ECASS) III extended the thrombolysis time window for patients with stroke from 3 to 4.5 hours after symptom onset. We investigated the effect of the extended thrombolysis time window on the proportion of recombinant tissue-type plasminogen activator-treated stroke patients and on the time of treatment initiation after hospital arrival.

Methods—

The present study was based on a prospective database of 93 hospitals of the Stroke Register of Northwestern Germany, which included 91 805 patients with ischemic stroke admitted between January 2007 and December 2009. Main outcome measures were the use of recombinant tissue-type plasminogen activator among patients with stroke and the door-to-needle time before and after the publication of ECASS III in September 2008 and subsequent changes of the German guidelines in May 2009.

Results—

Overall, 9262 patients (10.1%) were treated with recombinant tissue-type plasminogen activator. The proportion of thrombolyzed patients increased from 8.6% in 2007 to 11.7% in 2009. This increase was pronounced for patients admitted between 3 and 6 hours after symptom onset after the third quarter of 2008 (OR, 1.88; 95% CI, 1.24 to 2.85) and after the second and third quarters of 2009 (OR, 2.50; 95% CI, 1.69 to 3.69 and OR, 3.02; 95% CI, 2.07 to 4.41) compared with the first half year 2007. The proportion of patients with stroke with a door-to-needle time <60 minutes increased after publication of ECASS III (OR, 1.49; 95% CI, 1.37 to 1.63).

Conclusions—

Results of ECASS III were rapidly implemented in routine stroke care. Concerns of a delay in recombinant tissue-type plasminogen activator treatment initiation after the extension of the thrombolysis time window were not confirmed.
The Inclusion of Stroke in Risk Stratification for Primary Prevention of Vascular Events
Authors: Dhamoon Mandip S. , Moon Yeseon Park , Paik Myunghee C. , Sacco Ralph L. , Elkind Mitchell S.V.
Published: August 2011

Background and Purpose—

The Framingham coronary heart disease (CHD) risk score estimates 10-year risk of myocardial infarction (MI) and CHD death. Because preventive approaches to CHD and stroke are similar, a composite outcome may be more appropriate. We compared 10-year risk of (1) MI or CHD death; and (2) stroke, MI, or CHD death among individuals free of vascular disease.

Methods—

The Northern Manhattan Study contains a prospective, population-based study of stroke- and CHD-free individuals ≥40 years of age followed for a median of 10 years for vascular events. Framingham coronary heart disease risk score was calculated for each individual and for each category of predicted risk, Kaplan–Meier observed 10-year cumulative probabilities were calculated for (1) MI or CHD death; and (2) stroke, MI, or CHD death. The cumulative probability of (1) was subtracted from (2), and 95% CIs for the difference were obtained with 1000 bootstrap samples. Using stratified analyses by race–ethnicity, we compared risk differences among race–ethnic groups.

Results—

Among 2613 participants (53% Hispanic, 25% non-Hispanic black, and 20% non-Hispanic white), observed 10-year risk of MI or CHD death was 14.20%. With stroke in the outcome, observed risk was 21.98% (absolute risk difference, 7.78%; 95% CI, 5.86% to 9.75%). The absolute risk difference among blacks was significantly larger than among whites (
P
=0.01).

Conclusions—

In this multiethnic urban population, adding stroke to the risk stratification outcome cluster resulted in a 55% relative increase in estimated risk and crossing of the absolute risk threshold (>20% over 10 years) considered for preventive treatments such as statins.