bluuebunny/crossref_metadata_2025 huggingface dataset:
Mg2+- and MgATP-inhibited and Ca2+/calmodulin-sensitive TRPM7-like current in hepatoma and hepatocytes
Authors: Mishra Ravi , Rao Vidya , Ta Robert , Shobeiri Navid , Hill Ceredwyn E.
Published: August 2009

Although understood to be ubiquitously expressed, the functional identification and significance of Mg2+-inhibited, nonspecific cation currents has been established in only a few cell types. Here we identified an outwardly rectifying nonspecific cation current in quiescent rat hepatocytes and the proliferating and polarized rat hepatoma, WIF-B. Under whole cell recording conditions in which cells were bathed and dialyzed with Na-gluconate solutions, the latter Ca2+and Mg2+free, current reversed close to 0 mV, was time independent, and was greater than 10 times higher at +120 mV compared with −120 mV. Outward current at −120 mV developed slowly, from 17.7 ± 10.3 pA/pF at patch rupture to 106.6 ± 15.6 pA/pF at 12 min in WIF-B cells, and 4.9 ± 2.7 to 20.6 ± 5.6 pA/pF in rat hepatocytes. The nonspecific TRP channel inhibitor, 2-aminoethoxyphenylborate (2-APB), inhibited current (IC50= 72 ± 13 μM) and caused apoptotic cell death in WIF-B cells. Rat hepatocyte survival was more resistant to 2-APB. Dialysis of WIF-B cells with physiological concentrations of Mg2+and Mg-ATP, but not ATP alone, inhibited current development, suggesting that Trpm7 rather than Trpm6 underlies this current. RT-PCR demonstrated that both Trpm6 and Trpm7 are expressed at similar levels in both cell types, suggesting that the functional differences noted are not transcript dependent. Intracellular Ca2+(IC50= 125 ± 35 nM) also inhibited current development, and this could be partially relieved by the calmodulin and Ca2+/calmodulin-dependent kinase inhibitors W-7, staurosporine, KN-93, or calmodulin kinase II (CaMKII) inhibitory peptide. To summarize, our results show that in addition to their established Mg2+sensitivity, Trpm7-like channels are inhibited by cytosolic Ca2+in a CaMKII-dependent manner and may support hepatocellular survival during proliferation.
Synergistic effects of autologous cell and hepatocyte growth factor gene therapy for neovascularization in a murine model of hindlimb ischemia
Authors: Yamamoto Yasutaka , Matsuura Takashi , Narazaki Genta , Sugitani Miyoko , Tanaka Kohei , Maeda Akihiro , Shiota Goshi , Sato Kenzo , Yoshida Akio , Hisatome Ichiro
Published: August 2009

Autologous cell implantation and angiogenic gene therapy have been evaluated in critical limb ischemic patients. Here, we compared the features of these strategies individually and in combination. C57BL/6J mice with ischemic hindlimbs were injected with adherent mononuclear cells (aMNCs) from bone marrow or adenovirus encoding the hepatocyte growth factor (HGF) gene (Ad-HGF). Under comparable angiogenic conditions, 10 × 105 aMNCs produced significantly higher amounts of VEGF and FGF-2 and stimulated the number of arterioles in ischemic muscle compared with 1 × 108 plaque-forming units (pfu) of Ad-HGF. Ad-HGF produced 10 times more HGF in ischemic muscle compared with aMNCs. Injection of 0.3 × 105 aMNCs previously transfected with Ad-HGF (aMNC/Ad-HGF) increased blood flow and elevated the numbers of capillaries and arterioles to levels comparable with that seen with 10 × 105 aMNCs or 1 × 108 pfu of Ad-HGF. Hypoxic conditions induced the apoptotic death of aMNCs. However, coincubation with HGF or aMNC/Ad-HGF protected cells against apoptosis. HGF stimulated the migration of aMNCs, and the migration capacity of the aMNC/Ad-HGF group was significantly higher than that in the aMNC/Ad-LacZ group. In conclusion, cell-based HGF gene therapy decreased the number of cells required for neovascularization. This strategy can be an effective angiogenic therapy.
TNF-α inhibition attenuates adverse myocardial remodeling in a rat model of volume overload
Authors: Jobe Lynetta J. , Meléndez Giselle C. , Levick Scott P. , Du Yan , Brower Gregory L. , Janicki Joseph S.
Published: August 2009

Tumor necrosis factor (TNF)-α is a proinflammatory cytokine that has been implicated in the pathogenesis of heart failure. In contrast, we have recently shown that myocardial levels of TNF-α are acutely elevated in the aortocaval (AV) fistula model of heart failure. Based on these observations, we hypothesized that progression of adverse myocardial remodeling secondary to volume overload would be prevented by inhibition of TNF-α with etanercept. Furthermore, a principal objective of this study was to elucidate the effect of TNF-α inhibition during different phases of the myocardial remodeling process. Eight-week-old male Sprague-Dawley rats were randomly divided into the following three groups: sham-operated controls, untreated AV fistulas, and etanercept-treated AV fistulas. Each group was further subdivided to study three different time points consisting of 3 days, 3 wk, and 8 wk postfistula. Etanercept was administered subcutaneously at 1 mg/kg body wt. Etanercept prevented collagen degradation at 3 days and significantly attenuated the decrease in collagen at 8 wk postfistula. Although TNF-α antagonism did not prevent the initial ventricular dilatation at 3 wk postfistula, etanercept was effective at significantly attenuating the subsequent ventricular hypertrophy, dilatation, and increased compliance at 8 wk postfistula. These positive adaptations achieved with etanercept administration translated into significant functional improvements. At a cellular level, etanercept also markedly attenuated increases in cardiomyocyte length, width, and area at 8 wk postfistula. These observations demonstrate that TNF-α has a pivotal role in adverse myocardial remodeling and that treatment with etanercept can attenuate the progression to heart failure.
Myoglobin concentration in skeletal muscle fibers of chronic heart failure patients
Authors: Bekedam Martijn A. , van Beek-Harmsen Brechje J. , van Mechelen Willem , Boonstra Anco , van der Laarse Willem J.
Published: August 2009

The purpose of this study was to determine the myoglobin concentration in skeletal muscle fibers of chronic heart failure (CHF) patients and to calculate the effect of myoglobin on oxygen buffering and facilitated diffusion. Myoglobin concentration, succinate dehydrogenase (SDH) activity, and cross-sectional area of individual muscle fibers from the vastus lateralis of five control and nine CHF patients were determined using calibrated histochemistry. CHF patients compared with control subjects were similar with respect to myoglobin concentration: type I fibers 0.69 ± 0.11 mM (mean ± SD), type II fibers 0.52 ± 0.07 mM in CHF vs. type I fibers 0.70 ± 0.09 mM, type II fibers 0.49 ± 0.07 mM in control, whereas SDH activity was significantly lower in CHF in both fiber types ( P < 0.01). The myoglobin concentration in type I fibers was higher than in type II fibers ( P < 0.01). Consequently, the oxygen buffering capacity, calculated from myoglobin concentration/SDH activity was increased in CHF: type I fibers 11.4 ± 2.1 s, type II fibers 13.6 ± 3.9 s in CHF vs. type I fibers 7.8 ± 0.9 s, type II fibers 7.5 ± 1.0 s in control, all P < 0.01). The calculated extracellular oxygen tension required to prevent core anoxia (Po2crit) in muscle fibers was similar when controls were compared with patients in type I fibers 10.3 ± 0.9 Torr in CHF and 11.5 ± 3.3 Torr in control, but was lower in type II fibers of patients 6.1 ± 2.8 Torr in CHF and 14.7 ± 6.2 Torr in control, P < 0.01. The lower Po2crit of type II fibers may facilitate oxygen extraction from capillaries. Reduced exercise tolerance in CHF is not due to myoglobin deficiency.
Effects of aging on human skeletal muscle after immobilization and retraining
Authors: Suetta C. , Hvid L. G. , Justesen L. , Christensen U. , Neergaard K. , Simonsen L. , Ortenblad N. , Magnusson S. P. , Kjaer M. , Aagaard P.
Published: August 2009

Inactivity is a recognized compounding factor in sarcopenia and muscle weakness in old age. However, while the negative effects of unloading on skeletal muscle in young individuals are well elucidated, only little is known about the consequence of immobilization and the regenerative capacity in elderly individuals. Thus the aim of this study was to examine the effect of aging on changes in muscle contractile properties, specific force, and muscle mass characteristics in 9 old (61–74 yr) and 11 young men (21–27 yr) after 2 wk of immobilization and 4 wk of retraining. Both young and old experienced decreases in maximal muscle strength, resting twitch peak torque and twitch rate of force development, quadriceps muscle volume, pennation angle, and specific force after 2 wk of immobilization ( P < 0.05). The decline in quadriceps volume and pennation angle was smaller in old compared with young ( P < 0.05). In contrast, only old men experienced a decrease in quadriceps activation. After retraining, both young and old regained their initial muscle strength, but old had smaller gains in quadriceps volume compared with young, and pennation angle increased in young only ( P < 0.05). The present study is the first to demonstrate that aging alters the neuromuscular response to short-term disuse and recovery in humans. Notably, immobilization had a greater impact on neuronal motor function in old individuals, while young individuals were more affected at the muscle level. In addition, old individuals showed an attenuated response to retraining after immobilization compared with young individuals.
Dietary nitrate supplementation reduces the O2cost of low-intensity exercise and enhances tolerance to high-intensity exercise in humans
Authors: Bailey Stephen J. , Winyard Paul , Vanhatalo Anni , Blackwell Jamie R. , DiMenna Fred J. , Wilkerson Daryl P. , Tarr Joanna , Benjamin Nigel , Jones Andrew M.
Published: August 2009

Pharmacological sodium nitrate supplementation has been reported to reduce the O2cost of submaximal exercise in humans. In this study, we hypothesized that dietary supplementation with inorganic nitrate in the form of beetroot juice (BR) would reduce the O2cost of submaximal exercise and enhance the tolerance to high-intensity exercise. In a double-blind, placebo (PL)-controlled, crossover study, eight men (aged 19–38 yr) consumed 500 ml/day of either BR (containing 11.2 ± 0.6 mM of nitrate) or blackcurrant cordial (as a PL, with negligible nitrate content) for 6 consecutive days and completed a series of “step” moderate-intensity and severe-intensity exercise tests on the last 3 days. On days 4–6, plasma nitrite concentration was significantly greater following dietary nitrate supplementation compared with PL (BR: 273 ± 44 vs. PL: 140 ± 50 nM; P < 0.05), and systolic blood pressure was significantly reduced (BR: 124 ± 2 vs. PL: 132 ± 5 mmHg; P < 0.01). During moderate exercise, nitrate supplementation reduced muscle fractional O2extraction (as estimated using near-infrared spectroscopy). The gain of the increase in pulmonary O2uptake following the onset of moderate exercise was reduced by 19% in the BR condition (BR: 8.6 ± 0.7 vs. PL: 10.8 ± 1.6 ml·min−1·W−1; P < 0.05). During severe exercise, the O2uptake slow component was reduced (BR: 0.57 ± 0.20 vs. PL: 0.74 ± 0.24 l/min; P < 0.05), and the time-to-exhaustion was extended (BR: 675 ± 203 vs. PL: 583 ± 145 s; P < 0.05). The reduced O2cost of exercise following increased dietary nitrate intake has important implications for our understanding of the factors that regulate mitochondrial respiration and muscle contractile energetics in humans.
Arginase inhibition restores NOS coupling and reverses endothelial dysfunction and vascular stiffness in old rats
Authors: Kim Jae Hyung , Bugaj Lukasz J. , Oh Young Jun , Bivalacqua Trinity J. , Ryoo Sungwoo , Soucy Kevin G. , Santhanam Lakshmi , Webb Alanah , Camara Andre , Sikka Gautam , Nyhan Daniel , Shoukas Artin A. , Ilies Monica , Christianson David W. , Champion Hunter C. , Berkowitz Dan E.
Published: August 2009

There is increasing evidence that upregulation of arginase contributes to impaired endothelial function in aging. In this study, we demonstrate that arginase upregulation leads to endothelial nitric oxide synthase (eNOS) uncoupling and that in vivo chronic inhibition of arginase restores nitroso-redox balance, improves endothelial function, and increases vascular compliance in old rats. Arginase activity in old rats was significantly increased compared with that shown in young rats. Old rats had significantly lower nitric oxide (NO) and higher superoxide (O2−) production than young. Acute inhibition of both NOS, with NG-nitro-l-arginine methyl ester, and arginase, with 2( S)-amino- 6-boronohexanoic acid (ABH), significantly reduced O2− production in old rats but not in young. In addition, the ratio of eNOS dimer to monomer in old rats was significantly decreased compared with that shown in young rats. These results suggest that eNOS was uncoupled in old rats. Although the expression of arginase 1 and eNOS was similar in young and old rats, inducible NOS (iNOS) was significantly upregulated. Furthermore, S-nitrosylation of arginase 1 was significantly elevated in old rats. These findings support our previously published finding that iNOS nitrosylates and activates arginase 1 (Santhanam et al., Circ Res 101: 692–702, 2007). Chronic arginase inhibition in old rats preserved eNOS dimer-to-monomer ratio and significantly reduced O2− production and enhanced endothelial-dependent vasorelaxation to ACh. In addition, ABH significantly reduced vascular stiffness in old rats. These data indicate that iNOS-dependent S-nitrosylation of arginase 1 and the increase in arginase activity lead to eNOS uncoupling, contributing to the nitroso-redox imbalance, endothelial dysfunction, and vascular stiffness observed in vascular aging. We suggest that arginase is a viable target for therapy in age-dependent vascular stiffness.
Immunotherapy of Head and Neck Cancer: Current and Future Considerations
Authors: Rapidis Alexander D. , Wolf Gregory T.
Published: August 2009

Patients with head and neck squamous cell carcinoma (HNSCC) are at considerable risk for death, with 5-year relative survival rates of approximately 60%. The profound multifaceted deficiencies in cell-mediated immunity that persist in most patients after treatment may be related to the high rates of treatment failure and second primary malignancies. Radiotherapy and chemoradiotherapy commonly have severe acute and long-term side effects on immune responses. The development of immunotherapies reflects growing awareness that certain immune system deficiencies specific to HNSCC and some other cancers may contribute to the poor long-term outcomes. Systemic cell-mediated immunotherapy is intended to activate the entire immune system and mount a systemic and/or locoregional antitumor response. The delivery of cytokines, either by single cytokines, for example, interleukin-2, interleukin-12, interferon-, interferon-, or by a biologic mix of multiple cytokines, such as IRX-2, may result in tumor rejection and durable immune responses. Targeted immunotherapy makes use of monoclonal antibodies or vaccines. All immunotherapies for HNSCC except cetuximab remain investigational, but a number of agents whose efficacy and tolerability are promising have entered phase 2 or phase 3 development.
A Decentralised Task Mapping Approach for Homogeneous Multiprocessor Network‐On‐Chips
Authors: Zipf Peter , Sassatelli Gilles , Utlu Nurten , Saint-Jean Nicolas , Benoit Pascal , Glesner Manfred
Published: August 2009

We present a heuristic algorithm for the run‐time distribution of task sets in a homogeneous Multiprocessor network‐on‐chip. The algorithm is itself distributed over the processors and thus can be applied to systems of arbitrary size. Also, tasks added at run‐time can be handled without any difficulty, allowing for inline optimisation. Based on local information on processor workload, task size, communication requirements, and link contention, iterative decisions on task migrations to other processors are made. The mapping results for several example task sets are first compared with those of an exact (enumeration) algorithm with global information for a 3 × 3 processor array. The results show that the mapping quality achieved by our distributed algorithm is within 25% of that of the exact algorithm. For larger array sizes, simulated annealing is used as a reference and the behaviour of our algorithm is investigated. The mapping quality of the algorithm can be shown to be within a reasonable range (below 30% mostly) of the reference. This adaptability and the low computation and communication overhead of the distributed heuristic clearly indicate that decentralised algorithms are a favourable solution for an automatic task distribution.
Oxaprozin‐Induced Apoptosis on CD40 Ligand‐Treated Human Primary Monocytes Is Associated with the Modulation of Defined Intracellular Pathways
Authors: Montecucco Fabrizio , Bertolotto Maria , Ottonello Luciano , Quercioli Alessandra , Mach François , Dallegri Franco
Published: August 2009

The modulation of CD40L activity might represent a promising therapeutic target to reduce monocyte inflammatory functions in chronic diseases, such as rheumatoid arthritis. In the present study, we investigated the possible influence of nonsteroidal anti‐inflammatory drugs (NSAIDs) on CD40L‐induced monocyte survival. Monocytes were isolated from buffy coats by using Ficoll‐Percoll gradients. Monocyte apoptosis was evaluated by fluorescence microscopy on cytopreps stained with acridine orange or using flow cytometry analysis of Annexin‐V and Propidium Iodide staining. Akt and NF‐κB activation was assessed using western blot. Caspase 3 activity was determined spectrophotometrically. Among different NSAIDs, only oxaprozin dose‐dependently increased apoptosis of CD40L‐treated monocytes. Oxaprozin pro‐apoptotic activity was associated with the inhibition of CD40L‐triggered Akt and NF‐κB phosphorylation and the activation of caspase 3. In conclusion, our data suggest that oxaprozin‐induced apoptosis in CD40L‐treated human monocytes is associated with previously unknown cyclooxygenase (COX)‐independent pathways. These intracellular proteins might be promising pharmacological targets to increase apoptosis in CD40L‐treated monocytes.
The Impact of HIV and Malaria Coinfection: What Is Known and Suggested Venues for Further Study
Authors: Hochman Sarah , Kim Kami
Published: August 2009

HIV and malaria have similar global distributions. Annually, 500 million are infected and 1 million die because of malaria. 33 million have HIV and 2 million die from it each year. Minor effects of one infection on the disease course or outcome for the other would significantly impact public health because of the sheer number of people at risk for coinfection. While early population-based studies showed no difference in outcomes between HIV-positive and HIV-negative individuals with malaria, more recent work suggests that those with HIV have more frequent episodes of symptomatic malaria and that malaria increases HIV plasma viral load and decreases CD4+ T cells. HIV and malaria each interact with the host's immune system, resulting in a complex activation of immune cells, and subsequent dysregulated production of cytokines and antibodies. Further investigation of these interactions is needed to better define effects of coinfection.
Synthesis and Characterization of Organic‐Inorganic Nanocomposite Poly‐o‐anisidine Sn(IV) Arsenophosphate: Its Analytical Applications as Pb(II) Ion‐Selective Membrane Electrode
Authors: Khan Asif Ali , Habiba Umme , Khan Anish
Published: August 2009

Poly‐o‐anisidine Sn(IV) arsenophosphate is a newly synthesized nanocomposite material and has been characterized on the basis of its chemical composition, ion exchange capacity, TGA‐DTA, FTIR, X‐RAY, SEM, and TEM studies. On the basis of distribution studies, the exchanger was found to be highly selective for lead that is an environmental pollutant. For the detection of lead in water a heterogeneous precipitate based ion‐selective membrane electrode was developed by means of this composite cation exchanger as electroactive material. The membrane electrode is mechanically stable, with a quick response time, and can be operated over a wide pH range. The selectivity coefficients were determined by mixed solution method and revealed that the electrode is sensitive for Pb(II) in presence of interfering cations. The practical utility of this membrane electrode has been established by employing it as an indicator electrode in the potentiometric titration of Pb(II).
Ultrasonographic Study of Gallbladder Wall Thickness and Emptying in Cirrhotic Patients without Gallstones
Authors: Loreno Massimiliano , Travali Salvatore , Bucceri Anna Maria , Scalisi Giuseppe , Virgilio Carla , Brogna Alfio
Published: August 2009

Background and Aim. Gallbladder wall thickening and impaired contractility are currently reported in cirrhotic patients and often related to portal hypertension and hepatic failure. The purpose of this work was to evaluate, by ultrasonographic method, gallbladder wall thickness and gallbladder emptying after a standard meal in normal subjects and in patients with compensated liver cirrhosis without gallstones.Methods. Twenty-three patients with Child-Pugh class A liver cirrhosis and twenty healthy controls were studied. Gallbladder wall thickness (GWT), gallbladder fasting volume (FV), residual volume (RV), and maximum percentage of emptying (%E) were calculated. Measurements of mean portal velocity, portal vein flow, and serum albumin were performed too. Statistical analysis was assessed by Student's “ttest” for unpaired data.Results. GWT was  cm in cirrhotic patients and  cm in controls (). FV and RV were, respectively,   and   in cirrhotic patients,   and   in healthy volunteers (). %E was smaller in cirrhotics () as compared to controls (; ).Conclusions. In patients with compensated liver cirrhosis without gallstones gallbladder wall thickness is increased whereas its contractility is reduced. These early structural and functional alterations could play a role in gallstone formation in more advanced stages of the disease.
The Fluctuation Niche in Plants
Authors: Terradas Jaume , Peñuelas Josep , Lloret Francisco
Published: August 2009

Classical approaches to niche in coexisting plants have undervalued temporal fluctuations. We propose that fluctuation niche is an important dimension of the total niche and interacts with habitat and life-history niches to provide a better understanding of the multidimensional niche space where ecological interactions occur. To scale a fluctuation niche, it is necessary to relate environmental constrictions or species performance not only to the absolute values of the usual environmental and ecophysiological variables but also to their variances or other measures of variability. We use Mediterranean plant communities as examples, because they present characteristic large seasonal and interannual fluctuations in water and nutrient availabilities, along an episodic-constant gradient, and because the plant responses include a number of syndromes coupled to this gradient.
Characterization of Synonymous Codon Usage Bias in the Duck Plague Virus UL35 Gene
Authors: Cai Ming-Sheng , Cheng An-Chun , Wang Ming-Shu , Zhao Li-Chan , Zhu De-Kang , Luo Qi-Hui , Liu Fei , Chen Xiao-Yue
Published: August 2009

Objective: The aim was to identify the codon usage bias between the newly identified duck plague virus (DPV) UL35 gene (GenBank accession No. EF643558) and the UL35-like genes of 27 other reference herpesviruses. Methods: A comparative analysis of the codon usage bias of the 28 herpesviruses was performed by using the CodonW 1.4 program and CUSP (create a codon usage table) program of EMBOSS (The European Molecular Biology Open Software Suite). Results: The results showed obvious differences of the synonymous codon usage bias in the 28 herpesviruses indicated by the Codon Adaptation Index, effective number of codons (ENc), and the value of G + C content at the 3rd codon position. The codon usage pattern of the DPV UL35 gene was phylogenetically conserved and similar to that of the UL35-like genes of the avian α-herpesvirus, with a strong bias towards the codons with A and T at the 3rd codon position. A cluster analysis of codon usage pattern of the DPV UL35 gene with other reference herpesviruses demonstrated that the codon usage bias of the UL35 genes of the 28 herpesviruses had a very close relation with their gene function. The ENc-plot revealed that the genetic heterogeneity in the DPV UL35 gene and the 27 reference herpesviruses were constrained by G + C content, while gene length exerted relatively weaker influences. In addition, comparisons of the codon preferences in the UL35 gene of DPV with those of Escherichia coli, yeast and humans revealed that there were 33 codons showing distinct usage differences between the DPV and yeast, and 38 between the DPV and humans, but only 31 between the DPV and E. coli. Therefore, the E. coli system may be more suitable for the expression of the DPV UL35 gene. Conclusion: Together, these results may improve our understanding of the evolution, pathogenesis and functional studies of DPV, as well as contribute significantly to the area of herpesvirus research and possibly studies with other viruses.
Stroke Severity, Its Correlates and Impact on Thrombolysis in a Population-Based Study
Authors: Lyrer P.A. , Fluri F. , Gostynski M. , Bonati L. , Papa S. , Ajdacic-Gross V. , Engelter S.T.
Published: August 2009

Objective: Data about the distribution of stroke severity and its correlates are sparse. In a population-based approach, we determined the NIH Stroke Scale Score (NIHSSS) and studied associations with demographic variables, stroke unit care, etiology, the onset assessment interval (OAI), and the rate of thrombolysis. Methods: We performed a databank-based post-hoc analysis of data ascertained during the prospective, population-based stroke study among the 188,015 permanent residents of Basel City, Switzerland. Results: In 246/269 (91.4%) patients, NIHSSS was available. The median NIHSSS was 5.0 ± 6.0. NIHSSS 0–6, 7–15, and >15 were present in 156 (63%), 56 (23%), and 34 (14%) patients. Higher NIHSSS were associated with advancing age (p = 0.038), female gender (p = 0.04), stroke unit treatment (p = 0.003), cardioembolism (p < 0.001), shorter OAI (p = 0.009), and thrombolytic therapy (p < 0.001). In multivariate regression analyses, age, OAI, and thrombolysis correlated independently with higher NIHSSS. Stroke unit patients differed from non-stroke unit patients in shorter OAI, younger age, and higher NIHSSS. Conclusion: In a geographically defined stroke population, 1/3 patients had moderate-to-severe stroke. Patients with less severe strokes were younger, sought medical attention later and were less likely to receive thrombolysis. Thus, public stroke awareness programs might consider targeting also younger individuals and stress that also mild-to-moderate strokes benefit from emergency medical care.
PGE2 Causes Mesangial Cell Hypertrophy and Decreases Expression of Cyclin D3
Authors: Qian Qian , Kassem Kamal M. , Beierwaltes William H. , Harding Pamela
Published: August 2009

Background/Aims: Glomerular hypertrophy is a feature of many glomerular diseases and is associated with the development of renal failure. We previously demonstrated that the cyclooxygenase 2 inhibitor, NS398, reduced glomerular size after uninephrectomy. Thus, we hypothesized that prostaglandin (PG) E2 would cause mesangial cell hypertrophy in vitro. Methods: We used a mesangial cell line and primary culture of rat mesangial cells. The effects of PGE2 on mesangial cell hypertrophy were determined using immunohistochemistry and image analysis to assess cell size, 3H-leucine incorporation as a measure of protein synthesis and flow cytometry to assess cell cycle status. Western blot was used to examine the effect of PGE2 on expression of cyclin D3, p15, p27 and cyclin-dependent kinase 4 – known regulators of the cell cycle. Results: PGE2 increased cell size by 13% and protein synthesis (3H-leucine incorporation) by 35% over 24 h. By flow cytometry, PGE2 increased the percentage of cells in G0/G1 phase of the cell cycle from 70.13 ± 1.01 to 74.06 ± 1.18% and conversely, decreased the number of cells in S phase from 24.07 ± 1.06 to 22.03 ± 0.78%. The number of cells in G2/M was also reduced. Expression of cyclin D3 was decreased by 60% after treatment with PGE2, while expression of p27 was increased. The effects of PGE2 on cell size and flow cytometry were reproduced by the prostaglandin E2 receptors EP1/EP3 agonist sulprostone. Conclusion: PGE2 induces mesangial cell hypertrophy and cell cycle arrest via its EP1 receptor.
Optineurin and Its Mutants: Molecules Associated with Some Forms of Glaucoma
Authors: Chalasani Madhavi Latha , Swarup Ghanshyam , Balasubramanian Dorairajan
Published: August 2009

Aims: Optineurin is a gene (OPTN) associated with normal-tension glaucoma and primary open-angle glaucoma. The aim of our study is to understand the functions of the protein optineurin in retinal ganglion cells, and the modifications that come about in its functions when mutations occur in its sequence. Methods: We have worked with the rat retinal ganglion cell line RGC5, as well as with some others such as HeLa (human cancer cell line), COS-1 (monkey kidney cell line), IMR-32 (human neuroblastoma cell line), and D407 (human retinal pigment epithelial cell line). The cDNAs of wild-type OPTN, and of its mutants H26D, E50K, R454Q and H486R, were transfected into each of these cell lines and the effects studied. Localization of the proteins in cells was monitored by confocal microscopy and cell imaging, and their interaction with other cellular proteins by yeast two-hybrid assay and co-immunoprecipitation. Results: We found optineurin to interact with several proteins, which are involved in NF-κB regulation, vesicular trafficking at recycling endosomes, immune response and transcription regulation. Some of these proteins showed altered/reduced interactions with E50K and H486R mutants. E50K, a severe phenotype-related mutant of optineurin, was found to selectively kill RGC5 cells through oxytosis and apoptosis, while none of the other mutants did so. Conclusions: Our results indicate that several independent mechanisms are likely to be involved in the pathogenesis of glaucoma caused by mutations in optineurin.
A Comparative Study: Early Results and Complications of Percutaneous and Surgical Closure of Ventricular Septal Defect
Authors: Zheng Qijun , Zhao Zhijing , Zuo Jian , Yang Jian , Wang Hongbing , Yu Shiqiang , Yi Dinghua
Published: August 2009

Objective: Given the increasing popularity of percutaneous closure of ventricular septal defects (VSDs), a comparative analysis of the different treatments (surgical vs. percutaneous closure) for VSDs is needed. Methods: This is a single center, retrospective clinical study. A total of 2,178 patients with VSDs were enrolled, including 852 patients treated with percutaneous closure (device group) and 1,326 patients underwent traditional surgical repair (surgical group). Several characteristics (e.g. procedure success rate, complications, blood transfusions) were compared between the 2 groups. Results: There were no differences (p > 0.05) between the device and surgical groups according to the success rates (99.8 vs. 100%) and occurrences of main complications (1.9 vs. 2.5%). The incidence of minor complications was significantly lower in the device group (0.6%, 5/852) than the surgical group (6.4%, 85/1,326) (p < 0.01). No blood transfusions were needed in the device group, while 136 patients (10.3%) required blood transfusions in the surgical group. The duration of hospital stay was shorter in the device group than in the surgical group (3.2 vs. 12.9 days, p < 0.01). Conclusion: Percutaneous VSD closure is an effective method with fewer complications, shorter hospital stay and good cosmetic effect. It can serve as a reasonable alternative treatment for traditional open heart surgery.
The Immunoglobulin E-Toll-Like Receptor Network
Authors: Novak Natalija , Bieber Thomas , Peng Wen-Ming
Published: August 2009

Allergens and microbial antigens impact on effector cells and antigen-presenting cells in allergic diseases. Allergens bind specifically to immunoglobulin E (IgE) linked to the high-affinity receptor for IgE (FcεRI) and stimulate a cascade of cellular events. This leads to the release of mediators of allergic reactions by effector cells on the one hand and antigen uptake, presentation and T cell priming by antigen-presenting cells on the other hand. In contrast, microbial antigens are recognized by pattern-recognition receptors (PRRs) of the innate immune system, to which Toll-like receptors (TLRs) belong. In view of the high number of microbial antigens, allergens and other soluble ligands in the cellular microenvironment in vivo, it is very likely that not only separate, but also concomitant stimulation of both receptor types, i.e. FcεRI and TLRs, occurs frequently under physiological conditions and in particular in the context of allergic and infectious disorders. Thus, interaction of TLRs with FcεRI and regulation of the IgE synthesis is of critical immunological importance, since it might profoundly modify the activation state of cells and the nature of the evolving immune responses. Current knowledge about the cross talk of TLRs with FcεRI- and IgE-related immune responses is discussed herein.
Role of Interleukin-17A in the Eosinophil Accumulation and Mucosal Remodeling in Chronic Rhinosinusitis with Nasal Polyps Associated with Asthma
Authors: Saitoh T. , Kusunoki T. , Yao T. , Kawano K. , Kojima Y. , Miyahara K. , Onoda J. , Yokoi H. , Ikeda K.
Published: August 2009

Background: Interleukin (IL)-17A is a highly inflammatory cytokine with a robust effect on stromal cells in many tissues. Although IL-17A is known to be associated with inflammatory lung disorders by triggering an accumulation of neutrophils, the effect of IL-17A on the upper airway is still uncertain. The expression of IL-17A and its role were investigated in the nasal polyps of chronic rhinosinusitis associated with asthma. Methods: IL-17A was detected by immunohistochemistry and quantitative real-time RT-PCR. The cellular source of IL-17A was examined by double staining with EG2, CD4 and neutrophil elastase. The tissue remodeling of the nasal polyps was evaluated by assessing the epithelial damage and basement membrane thickness. Results: Both the immunoreactivity and mRNA of IL-17A were significantly detected in the nasal polyps in comparison with control normal sinus mucosa. The localization of IL-17A expression predominantly coincided with eosinophils and CD4-positive lymphocytes. Furthermore, the number of IL-17A-positive cells correlated with tissue eosinophils, but not with neutrophils. The degree of epithelial damage and basement membrane thickness was dependent on the number of infiltrated IL-17A-positive cells. Conclusion: The present study suggests, for the first time, that IL-17A plays an important role in the eosinophil accumulation in the nasal polyps and the remodeling of the nasal polyps of chronic rhinosinusitis associated with asthma.
Characterization of Anti-Inflammatory Properties and Evidence for No Sedation Liability for the Novel Antihistamine SUN-1334H
Authors: Mandhane Sanjay N. , Shah Jigar H. , Bahekar Prashant C. , Mehetre Sameer V. , Pawar Chandrashekhar A. , Bagad Ashish S. , Chidrewar Gajanan U. , Rao Chitturi Trinadha , Rajamannar Thennati
Published: August 2009

Background: The anti-inflammatory potential of antihistamines has significant clinical utility. Long-term pharmacotherapy of so-called ‘safe’ antihistamines may be hampered by side effects in the central nervous system. In the present study, the new potential antihistamine SUN-1334H was compared with different antihistamines for anti-inflammatory effects, sedation potential and interaction with alcohol. Method: Nasal and skin allergy were induced in guinea pig and mice by ovalbumin sensitization and challenge. Neurogenic nasal inflammation was induced by capsaicin. Sedation potential and interaction with alcohol were assessed by i.v. and intracerebroventricular pentobarbital-induced sedation and alcohol-induced ataxia models. Results: Ovalbumin sensitization and challenge caused rhinitis pathology including inflammatory cell infiltration, IL-4, and protein leakage in the nasal lavage fluid (NLF) and presence of inflammatory cells in nasal epithelium. A 5-day treatment of antihistamines reduced these markers of inflammation. SUN-1334H, cetirizine and hydroxyzine caused comparable inhibition of NLF leukocytes, IL-4 and total protein concentrations. Fexofenadine and desloratadine showed moderate inhibition of NLF leukocytes and had no significant effect on IL-4 concentration. While fexofenadine had no effect on total protein concentration, the effect of desloratadine was comparable with the other antihistamines. In neurogenic nasal inflammation induced by capsaicin, SUN-1334H and fexofenadine caused better inhibition at lower and middle dose levels than the other antihistamines. In skin allergy models, SUN-1334H showed potent reduction of passive and active cutaneous anaphylactic reactions. In central nervous system side effects models, SUN-1334H, desloratadine and fexofenadine were devoid of any significant effects. Conclusions: The results are suggestive of a high anti-inflammatory to sedation index of SUN-1334H among leading antihistamines.
Impaired Glucose Metabolism in Women with Polycystic Ovary Syndrome
Authors: Vrbikova Jana , Fanta Michal , Cibula David , Vondra Karel , Bendlova Bela
Published: August 2009

Background: To compare the prevalence of impaired glucose tolerance (IGT) and type 2 diabetes mellitus (T2DM) in lean and overweight/obese women with polycystic ovary syndrome (PCOS), with the data from a normal population sample. Methods: PCOS-affected women fulfilling ESHRE diagnostic criteria underwent an oral glucose tolerance test. Prevalence of IGT and T2DM in control sample of white healthy females was extracted from the published data from NHANES II. Results: In 225 women with PCOS, IGT was present in 6/104 (5.8%) lean and in 15/121 (12.4%) overweight/obese women. T2DM was present in 1/104 (1.0%) lean and in 3/121 (2.5%) overweight/obese PCOS women. In a sample of 643 women from NHANES II, the crude rate of IGT was 5.9%. IGT was significantly more common only in the overweight/obese PCOS subgroup as compared to the NHANES II cohort (χ2 = 5.99, p < 0.01). Conclusions: IGT was found significantly more frequently only in overweight/obese PCOS subjects in comparison with healthy controls.
Effect of Modafinil on Subjective Fatigue in Multiple Sclerosis and Stroke Patients
Authors: Brioschi Andrea , Gramigna Sandrine , Werth Esther , Staub Fabienne , Ruffieux Christiane , Bassetti Claudio , Schluep Myriam , Annoni Jean-Marie
Published: August 2009

Background: Modafinil has anecdotal response to neurological fatigue, but such an effect may depend on the type and location of cerebral impairment. Objectives: It was the aim of this study to compare fatigue observed in different neurological pathologies, to evaluate the tolerability to modafinil, and to describe changes in subjective fatigue. Methods: We enrolled 14 brainstem or diencephalic stroke (BDS) patients, 9 cortical stroke (CS) patients and 17 multiple sclerosis (MS) patients. The Fatigue Assessment Instrument severity scale was performed at baseline, after 3 months of modafinil and after 1 month of washout. Cognition, mood and somnolence were assessed. A subgroup of 14 patients underwent activity measures before and during treatment. Results: Thirty-one patients completed the study (10 BDS, 9 CS, 12 MS). The responder profile is more frequent in MS than in CS (p = 0.04), and in BDS than in CS patients (p = 0.04). Actiwatch measures showed no changes in activity during, before and after therapy. Conclusion: Modafinil was tolerated in 75% of patients at small doses and seemed to improve the severity of fatigue in the MS and BDS groups but not in the CS group. There was no modification in measured physical activity.
The Potential Anti-Inflammatory Effect of Exercise in Chronic Obstructive Pulmonary Disease
Authors: van der Vlist Jantine , Janssen Thomas W.J.
Published: August 2009

People with chronic obstructive pulmonary disease (COPD) suffer from muscle dysfunction which seems to be partly caused by systemic inflammation. Muscle protein breakdown as well as synthesis might be affected by this systemic inflammation. Additionally, it seems to induce excessive oxidative stress and reduce the level of growth-stimulating factors. As exercise training can have an anti-inflammatory effect in healthy people, the main question in this review is whether exercise (training) can also induce such effects in patients with COPD. However, because of the known inflammatory response after an acute bout of exercise, some researchers are afraid that exercise might actually worsen the inflammation in COPD. Recent evidence suggests, however, that the response might actually be anti-inflammatory and thus beneficial. Unfortunately, the evidence about the response of the inflammatory cytokines tumor necrosis factor-α and interleukin-6 to exercise in patients with COPD is inconsistent, making it impossible to conclude whether a single exercise bout is harmful or beneficial in patients with COPD. Long-term exercise training in healthy people as well as in patients with chronic heart failure, another chronic inflammatory disease, seems to have beneficial effects on the inflammatory response. In patients with COPD, however, no training-induced changes in cytokine levels have been found and it must be concluded that physical exercise training does not seem to have an anti-inflammatory effect in COPD. On the other hand, it does not have a proinflammatory effect, and since patients with COPD benefit from exercise training with regard to other health parameters it is still recommended that they exercise regularly.
Characterization of a Factor from Rat Serum with Blocking Activity in the Passive Cutaneous Anaphylactic Reaction
Authors: Theobald K. , König W.
Published: August 2009

A blocking factor (BF) was obtained from normal and rat reaginic serum which inhibited the passive cutaneous anaphylactic (PCA) reaction of mouse and rat IgE in rat skin. Several chromatographic procedures on DEAE- and CM-cellulose at various pH values were compared in an attempt to obtain BF with high activity. BF-containing fractions after DEAE chromatography at pH 5,4 revealed two components on SDS-polyacrylamide gel: one at 50,000–60,000 and the other at 65,000–70,000 daltons. The latter component was also obtained when normal or rat reaginic serum was adsorbed on Sepharose anti rat IgG. The eluate inhibited the PCA reaction and demonstrated two bands on SDS-polyacrylamide gel, one at 150,000 (IgG) and another at 65,000–70,000 daltons. Purified IgG and rat albumin did not inhibit the PCA reaction. Antisera against BF were obtained with the component at 65,000–70,000 daltons. Anti-BF revealed a specificity for two bands in the cathodal position when assayed against normal rat serum; one of which was IgG. Anti-BF recognized one component in the DEAE fraction when analyzed against the DEAE fraction which contained blocking activity. BF is immunologically distinct from IgG. After immunoadsorption of normal or rat reaginic serum on Sepharose anti-BF columns, an eluate was obtained which showed blocking activity in the PCA reaction and two components on SDS-polyacrylamide gel, one at 150,000 and the other at 65,000–70,000 daltons. Evaluation of blocking factor by isoelectric focussing obtained after various procedures revealed an isoelectric point of 4.7. BF shows dual functions: it inhibits and at high dilutions also enhances the PCA reaction. It is suggested that this factor has a modulatory role in the allergic disease process.
Detection and Characterization of Polymers in Cephalothin by Passive Cutaneous Anaphylaxis in Mice
Authors: Iwata Masayuki , Tokiwa Hiroshi , Matuhasi Tyoku
Published: August 2009

Immunization of BALB/c mice by repeated injections of cephalothin (CET) – Ascaris suum extract conjugate resulted in formation of IgE antibodies, which were able to sensitize syngeneic animals for passive cutaneous anaphylaxis (PCA). By means of high-pressure liquid chromatography, a fraction with extremely high PCA-eliciting activity, but without appreciable antimicrobial activity, was isolated from the CET preparation. Physicochemical analyses of the fraction identified the major component of polymer impurities as being a proteinaceous complex with a molecular weight of 6,580. Very little cross-reactivity of CET and potassium benzyl penicillin (PcG) was noted when these antibiotics were used as the challenge antigens for PCA induced by corresponding murine antisera. The results of the inhibition studies indicated, however, that at least two antigens were involved in the PCA induced by anti-CET antibodies, one strictly specific for CET and another shared by PcG. Evidence was also presented that the nucleus structure and acyl side-chain structure of CET play the major role in the PCA elicited by the challenge with CET and its polymer, respectively.
Studies in Immunodermatology
Authors: Dabski Krzysztof , Beutner Ernst H. , Jablonska Stefania
Published: August 2009

Normal human sera contain stratum corneum (SC) antibodies to two biochemically distinct antigens of normal skin as demonstrated by indirect immunofluorescence (IF) staining with selectively absorbed sera. Absorption of normal human sera with ground callus removed the SC antibodies reactive with the horny layer of normal skin but did not affect the SC antibody titers on cut edges of trypsin-digested callus or lesions of monkey skin induced by scratching. Conversely, absorption of the same sera with a carbohydrate-type SC antigen reduced SC antibody titers on the cut edges of trypsin-digested callus and on lesions of monkey skin induced by scratching but did not alter titers of SC antibodies on normal skin sections.
Factors Influencing Bonding of Bromelain Agglutinators and Their Fab Fragments
Authors: Waller Marion , Richard Alfred J.
Published: August 2009

The complex of bromelain agglutinators and their homologous Fab fragments is dissociated by gel chromatography under certain conditions. When albumin is present as a source of thiol groups, Fab fragments previously treated with N-ethylmaleimide (NEM) will dissociate from the agglutinators (fluid phase). If anti-Rh Fab fragments are bound to Rh-positive erythrocytes, the agglutinates are not dissociated by thiols (cellular phase). Prior to erythrocyte sensitization, the agglutinator site on Fab fragments can be blocked by thioldisulfide exchange. Once the Fab fragments are coated on erythrocytes, the agglutinator site is more readily available than it was prior to sensitization, as evidenced by inhibition with 0.01 M NEM. The differences between the bonding characteristics of the fluid phase and the cellular phase and the influence of mercaptoalbumin on the agglutinator-Fab complex suggest that the agglutinators are not antibodies.
Detection of Complexed IgE in Sera of Patients Allergic to the Pollen of Parietaria officinalis
Authors: Oreste U. , Calligaris V. , Ruffilli A.
Published: August 2009

34 sera of patients allergic to the pollen of Parietaria officinalis were analyzed by ultracentrifugation on sucrose gradient (40–10%) in 0.9% NaCl at 24,000 rpm with a SW 27.1 rotor for complexed circulating IgE. Total and specific anti-PO IgE content of the gradient’s fractions were determined by radioimmunoassay. IgE presence was demonstrated in at least two peaks; one corresponding to monomeric IgE, another corresponding to IgE with a sedimentation constant higher than that of the monomers. The ratio between monomeric and complexed IgE was different for each serum. Discordant results of PRIST and RIST analyses suggest that complexed IgE has fewer sites available for reaction with anti-IgE than the monomers.