bluuebunny/crossref_metadata_2025 huggingface dataset:
Pharmacokinetics of Puerarin and Ginsenoside Rg1 of CBN Injection and the Relation with Platelet Aggregation in Rats
Authors: Liu Ruining , Xing Dongming , Lu Hong , Wu Hao , Du Lijun
Published: December 2006

In order to study the pharmacokinetics of puerarin and ginsenoside Rg1 of cerebral blood nutrition (CBN) and its relationship with pharmacodynamics of platelet aggregation induced by ADP in rat, the blood samples after injection were collected. The concentrations of puerarin and ginsenoside Rg1 in plasma were determined by RP-HPLC, and the platelet aggregations were observed simultaneously. The data showed that there was distinct statistic significance ( p < 0.01) for puerarin processing, which was a single compartment model with quick elimination rate ( t 1/2β = 18 min ) and MRT (26 min), while ginsenoside Rg1 processing was a double compartment model with rapid distribution rate ( t 1/2α = 8 min ), slow elimination rate ( t 1/2β = 11 hours ) and MRT (3.3 hours). Effects of anti-platelet aggregation were presented at 5–10 min, 45–90 min and 6–8 hours after injection separately, and the corresponding concentrations of puerarin were 25–21 μg/ml, 4.5–0.8 μg/ml and 0 μg/ml, ginsenoside Rg1 were 7.6–6.7 μg/ml, 1.2–0.6 μg/ml and 1.8–0.5 μg/ml. The two components presented a positive correlation between their concentrations and the effect of anti-platelet aggregation in 5–10 min after CBN injection (coefficient of correlation were 0.999 and 0.995). However, it was noted that the effect was still stronger even when concentrations of puerarin and ginsenoside Rg1 in plasma decreased. Therefore, puerarin and ginsenoside Rg1 not only had different pharmacokinetics, but also had a positive correlation with platelet aggregation, just in 5–10 min after CBN injection.
Rehmannia glutinosaActivates Intracellular Antioxidant Enzyme Systems in Mouse Auditory Cells
Authors: Yu Hyeon-Hee , Kim Yeon-Hwa , Jung Su-Young , Shin Mee-Kyung , Park Rae-Kil , So Hong-Seob , Kim Ki-Young , Lee Da-Hong , You Yong-Ouk
Published: December 2006

Steamed roots of Rehmannia glutinosa (R. glutinosa) have been traditionally used in Oriental medicine for the treatment of auditory diseases such as tinnitus and hearing loss. To investigate whether the ethanol extract of steamed roots of R. glutinosa (SRG) increases activity of antioxidant enzymes and the level of glutathione (GSH), we measured activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), and glutathione reductase (GR) and GSH level in HEI-OC1 cells after treatment with 5–50 μg/ml of SRG. The SOD and CAT activities were significantly increased in the presence of SRG compared to the control group. Maximal activities of SOD and CAT were observed in these cells exposed to 10 μg/ml of SRG. The GPX activity also increased dramatically in response to the treatment with SRG in a dose-dependent manner. The GR activity was only increased in the presence of 50 μg/ml of SRG compared to the control group. The level of GSH gradually increased in the presence of 5–50 μg/ml of SRG. In the cytotoxicity test, 5–50 μg/ml of SRG did not show any significant cytotoxicity. These results suggest that the traditional use of R. glutinosa for the treatment of auditory diseases may be explained, in part, by activation of intracellular antioxidant enzyme systems. Further studies are necessary to clarify the active constituents of SRG responsible for such biomolecular activities.
SOME PROPERTIES OF A CLASS OF MERIT FUNCTIONS FOR SYMMETRIC CONE COMPLEMENTARITY PROBLEMS
Authors: LIU YONG-JIN , ZHANG LI-WEI , WANG YIN-HE
Published: December 2006

In this paper, we extend a class of merit functions proposed by Kanzow et al. (1997) for linear/nonlinear complementarity problems to Symmetric Cone Complementarity Problems (SCCP). We show that these functions have several interesting properties, and establish a global error bound for the solution to the SCCP as well as the level boundedness of every merit function under some mild assumptions. Moreover, several functions are demonstrated to enjoy these properties.
AN η-APPROXIMATION APPROACH IN NONLINEAR VECTOR OPTIMIZATION WITH UNIVEX FUNCTIONS
Authors: ANTCZAK TADEUSZ
Published: December 2006

In this paper, the so-called η-approximation approach is used to obtain the sufficient conditions for a nonlinear multiobjective programming problem with univex functions with respect to the same function η. In this method, an equivalent η-approximated vector optimization problem is constructed by a modification of both the objective and the constraint functions in the original multiobjective programming problem at the given feasible point. Moreover, to find the optimal solutions of the original multiobjective problem, it sufficies to solve its associated η-approximated vector optimization problem. Finally, the description of the η-approximation algorithm for solving a nonlinear multiobjective programming problem involving univex functions is presented.
CAPACITY ALLOCATION AND INVENTORY POLICY IN A DISTRIBUTION SYSTEM
Authors: DAI YUE , CHAO XIULI , FANG SHU-CHERNG , NUTTLE HENRY L. W.
Published: December 2006

We consider a single-period distribution system with one supplier and two retailers. The supplier may have infinite or finite capacity. The demand at each retailer is random. When a stockout occurs at one retailer the customer may go to the other retailer. We study both the decentralized and centralized inventory control problems. For the decentralized problem we show that a unique Nash equilibrium exists when the capacity at the supplier is infinite. However, when the capacity is finite, only under certain conditions does the Nash equilibrium exist. For centralized inventory control we obtain the optimal allocation that maximizes the expected profit of the entire supply chain. For the case of decentralized controls we also design channel coordination mechanisms, i.e., a decentralized cost structure resulting in a Nash equilibrium with chain-wide profits equal to those achieved under a fully centralized system. We compare the performance of two retailers in the decentralized and centralized controls and analyze the impact of channel coordination on the whole supply chain.
REVIEW OF LONG-TERM WARRANTY POLICIES
Authors: RAHMAN ANISUR , CHATTOPADHYAY GOPINATH
Published: December 2006

A large number of products are now being sold with long-term warranty policies. This is mainly due to fierce competition in the market and customer demand. Offering long-term warranty results in additional complexities in estimating costs associated with warranty servicing due to the uncertainties associated with usage, preventive maintenance and warranty servicing strategies. This paper reviews various long-term warranty policies and mathematical models for estimation of warranty servicing costs.
NEW OPTIMIZATION MODELS FOR DATA MINING
Authors: GLOVER FRED W. , KOCHENBERGER GARY
Published: December 2006

In recent years modern methods of optimization have contributed greatly to the advances in data mining and related areas. These contributions continue today and promise to further advance the state of the art both in terms of modeling innovations and new solution methodologies. In this paper, we present a new modeling and solution methodology for unsupervised clustering. Preliminary computational experience is given to illustrate the approach. This methodology is part of our current research and offers considerable opportunity for additional investigation to be conducted by other researchers.
WORKING KNOWLEDGE MINING BY PRINCIPLES FOR DEEP KNOWLEDGE
Authors: YU P. L.
Published: December 2006

We usually use a set of ideas, thinking paradigms and judgment rules, including alternatives, criteria, outcomes, preferences, to make decision. This set, known as actual domain (working knowledge) of habitual domain, will be stabilized over time unless extraordinary events occur. As such, our working knowledge cannot be broad and deep. Inevitably, we could get into decision traps, which lead us to making wrong decision or solving wrong problems. The actual domain is only a small part of our potential domain, the collection of all thoughts, ideas, thinking paradigms, etc. that have ever been encoded in our brain. In this paper, we will describe nine principles for deep knowledge, so that, we could expand and enrich our working knowledge by utilizing the potential domains of ourselves and other participants in the decision making. As a consequence, good ideas for solving challenging decision problems can be obtained or created. These principles are: The deep and down principle, the alternating principle, the contrasting and complementing principle, the revolving and cycling principle, the inner connection principle, the changing and transforming principle, the contradiction principle, the cracking and ripping principle, the void principle.
ANALYSIS OF THE SERVICE SECTOR
Authors: BERG DANIEL
Published: December 2006

This composite paper focuses on the Service Sector and its growing importance. Some of the characterizations made of the Service Sector using a simple analysis of data bases derived elsewhere are indicated. Discussion of the distinction between the role of intellectual property in the Goods Sector and the Service Sector is made.
FROM DATA MINING TO BEHAVIOR MINING
Authors: CHEN ZHENGXIN
Published: December 2006

Knowledge economy requires data mining be more goal-oriented so that more tangible results can be produced. This requirement implies that the semantics of the data should be incorporated into the mining process. Data mining is ready to deal with this challenge because recent developments in data mining have shown an increasing interest on mining of complex data (as exemplified by graph mining, text mining, etc.). By incorporating the relationships of the data along with the data itself (rather than focusing on the data alone), complex data injects semantics into the mining process, thus enhancing the potential of making better contribution to knowledge economy. Since the relationships between the data reveal certain behavioral aspects underlying the plain data, this shift of mining from simple data to complex data signals a fundamental change to a new stage in the research and practice of knowledge discovery, which can be termed as behavior mining. Behavior mining also has the potential of unifying some other recent activities in data mining. We discuss important aspects on behavior mining, and discuss its implications for the future of data mining.
ELICITING KNOWLEDGE ABOUT VALUES FOR PUBLIC POLICY DECISIONS
Authors: KEENEY RALPH L.
Published: December 2006

To solve or resolve public policy problems requires decisions. The intent is to choose the best alternative in each situation. Hence, the way the notion of best is defined and measured in each case is critical. A structured process is outlined to acquire knowledge about public values that could be used to evaluate alternatives and identify the best ones. These values are the foundation for any policy decision. The uses of these values for defining interest in the problem, creating alternatives, and identifying information needs are discussed. Research needs are outlined.
A NEW SYMMETRIC TWO-DIMENSIONAL ALGORITHM
Authors: AYUSO PEDRO FORTUNY , SCHWEIGER FRITZ
Published: December 2006

Continued fractions are deeply related to Singularity Theory, as the computation of the Puiseux exponents of a plane curve from its dual graph clearly shows. Another closely related topic is Euclid's Algorithm for computing the gcd of two integers (see [2] for a detailed overview). In the first section, we describe a subtractive algorithm for computing the gcd of n integers, related to singularities of curves in affine n-space. This gives rise to a multidimensional continued fraction algorithm whose version in dimension 2 is the main topic of the paper.
ON QUALIFIED RESIDUE DIFFERENCE SETS
Authors: BYARD KEVIN
Published: December 2006

Qualified residue difference sets of power n are known to exist for n = 2,4,6, as do similar sets that include the zero element. Both classes of sets are proved non-existent for n = 8.
SOME NUMBER THEORETIC APPLICATIONS OF A GENERAL MODULAR RELATION
Authors: KANEMITSU SHIGERU , TANIGAWA YOSHIO , TSUKADA HARUO
Published: December 2006

We state a form of the modular relation in which the functional equation appears in the form of an expression of one Dirichlet series in terms of the other multiplied by the quotient of gamma functions and illustrate it by some concrete examples including the results of Koshlyakov, Berndt and Wigert and Bellman.
Molecular Motors: A Theorist's Perspective
Authors: Kolomeisky Anatoly B. , Fisher Michael E.
Published: December 2006

Individual molecular motors, or motor proteins, are enzymatic molecules that convert chemical energy, typically obtained from the hydrolysis of ATP (adenosine triphosphate), into mechanical work and motion. Processive motor proteins, such as kinesin, dynein, and certain myosins, step unidirectionally along linear tracks, specifically microtubules and actin filaments, and play a crucial role in cellular transport processes, organization, and function. In this review some theoretical aspects of motor-protein dynamics are presented in the light of current experimental methods that enable the measurement of the biochemical and biomechanical properties on a single-molecule basis. After a brief discussion of continuum ratchet concepts, we focus on discrete kinetic and stochastic models that yield predictions for the mean velocity, V(F, [ATP], …), and other observables as a function of an imposed load force F, the ATP concentration, and other variables. The combination of appropriate theory with single-molecule observations should help uncover the mechanisms underlying motor-protein function.
Spectral and Dynamical Properties of Multiexcitons in Semiconductor Nanocrystals
Authors: Klimov Victor I.
Published: December 2006

Because of the strong spatial confinement of electronic wave functions and reduced dielectric screening, the effects of carrier-carrier Coulomb interactions are greatly enhanced in semiconductor nanocrystals (NCs) compared with those in bulk materials. These interactions open a highly efficient decay channel via Auger recombination, which represents a dominant recombination pathway for multiexcitons in NCs. Furthermore, strong Coulomb coupling between charge carriers leads to extremely efficient direct photogeneration of multiexcitons by single photons via carrier (or exciton) multiplication. This review focuses on spectral and dynamical properties of multiexcitons in semiconductor NCs. The specific topics discussed here include the structure of NC electronic states, spectral signatures of multiexcitons in transient absorption and photoluminescence, exciton-exciton interaction energies, Auger recombination, and carrier multiplication. This chapter also briefly reviews the implications of multiexciton effects for practical technologies, such as NC lasing and photovoltaics.
Resistance to high-fat diet in the female progeny of obese mice fed a control diet during the periconceptual, gestation, and lactation periods
Authors: Gallou-Kabani Catherine , Vigé Alexandre , Gross Marie-Sylvie , Boileau Catherine , Rabes Jean-Pierre , Fruchart-Najib Jamilla , Jais Jean-Philippe , Junien Claudine
Published: December 2006

With the worldwide epidemic of metabolic syndrome (MetS), the proportion of women that are overweight/obese and overfed during pregnancy has increased. The resulting abnormal uterine environment may have deleterious effects on fetal metabolic programming and lead to MetS in adulthood. A balanced/restricted diet and/or physical exercise often improve metabolic abnormalities in individuals with obesity and type 2 diabetes mellitus (T2D). We investigated whether reducing fat intake during the periconceptual/gestation/lactation period in mothers with high-fat diet (HFD)-induced obesity could be used to modify fetal/neonatal MetS programming positively, thereby preventing MetS. First generation (F1) C57BL/6J female mice with HFD-induced obesity and T2D were crossed with F1 males on control diet (CD). These F1 females were switched to a CD during the periconceptual/gestation/lactation period. At weaning, both male and female second generation (F2) mice were fed a HFD. Weight, caloric intake, lipid parameters, glucose, and insulin sensitivity were assessed. Sensitivity/resistance to the HFD differed significantly between generations and sexes. A similar proportion of the F1 and F2 males (80%) developed hyperphagia, obesity, and T2D. In contrast, a significantly higher proportion of the F2 females (43%) than of the previous F1 generation (17%) were resistant ( P < 0.01). Despite having free access to the HFD, these female mice were no longer hyperphagic and remained lean, with normal insulin sensitivity and glycemia but mild hypercholesterolemia and glucose intolerance, thus displaying a “satiety phenotype.” This suggests that an appropriate dietary fatty acid profile and intake during the periconceptual/gestation/lactation period helps the female offspring to cope with deleterious intrauterine conditions.
Cardiac matrix metalloproteinase-2 expression independently induces marked ventricular remodeling and systolic dysfunction
Authors: Bergman Marina R. , Teerlink John R. , Mahimkar Rajeev , Li Luyi , Zhu Bo-Qing , Nguyen Anita , Dahi Sia , Karliner Joel S. , Lovett David H.
Published: December 2006

Although enhanced cardiac matrix metalloproteinase (MMP)-2 synthesis has been associated with ventricular remodeling and failure, whether MMP-2 expression is a direct mediator of this process is unknown. We generated transgenic mice expressing active MMP-2 driven by the α-myosin heavy chain promoter. At 4 mo MMP-2 transgenic hearts demonstrated expression of the MMP-2 transgene, myocyte hypertrophy, breakdown of Z-band registration, lysis of myofilaments, disruption of sarcomere and mitochondrial architecture, and cardiac fibroblast proliferation. Hearts from 8-mo-old transgenic mice displayed extensive myocyte disorganization and dropout with replacement fibrosis and perivascular fibrosis. Older transgenic mice also exhibited a massive increase in cardiac MMP-2 expression, representing recruitment of endogenous MMP-2 synthesis, with associated expression of MMP-9 and membrane type 1 MMP. Increases in diastolic [control (C) 33 ± 3 vs. MMP 51 ± 12 μl; P = 0.003] and systolic (C 7 ± 2 vs. MMP 28 ± 14 μl; P = 0.003) left ventricular (LV) volumes and relatively preserved stroke volume (C 26 ± 4 vs. MMP 23 ± 3 μl; P = 0.16) resulted in markedly decreased LV ejection fraction (C 78 ± 7% vs. MMP 48 ± 16%; P = 0.0006). Markedly impaired systolic function in the MMP transgenic mice was demonstrated in the reduced preload-adjusted maximal power (C 240 ± 84 vs. MMP 78 ± 49 mW/μl2; P = 0.0003) and decreased end-systolic pressure-volume relation (C 7.5 ± 1.5 vs. MMP 4.7 ± 2.0; P = 0.016). Expression of active MMP-2 is sufficient to induce severe ventricular remodeling and systolic dysfunction in the absence of superimposed injury.
Calcium-mediated triggered activity is an underlying cellular mechanism of ectopy originating from the pulmonary vein in dogs
Authors: Hirose Masamichi , Laurita Kenneth R.
Published: December 2006

Paroxysmal atrial fibrillation associated with focal ectopy originating from the pulmonary vein (PV) can be preceded by variations in autonomic tone; however, the underlying cellular mechanisms are not clear. To determine the mechanisms of autonomically mediated PV ectopy, high-resolution optical mapping techniques were used to measure action potentials and Ca2+transients from the PV and the ligament of Marshall area in the arterially perfused canine left atrium. Rapid pacing was used to initiate ectopic activity during pituitary adenylate cyclase-activating polypeptide (PACAP) injection (1 nmol), as a surrogate for autonomic imbalance, before ( n = 9) and after ( n = 6) verapamil (10 nmol) administration. In all preparations, spontaneous activity was absent before rapid pacing. During PACAP injection, rapid pacing induced ectopic activity in eight of nine preparations. In contrast, before PACAP injection, rapid pacing did not induce ectopic activity. Activation maps of each episode of ectopic activity indicated that the site of origin occurred more frequently in the PV (70%) than in the ligament of Marshall (30%) area. As rapid pacing cycle length increased, so did the ectopic beat coupling interval. In addition, PACAP-induced ectopic activity was associated with large Ca2+transient amplitudes and was always suppressed by verapamil, a Ca2+channel blocker ( P < 0.05). Finally, during PACAP injection in the absence of an ectopic beat, spontaneous Ca2+release and delayed afterdepolarizations were observed simultaneously after termination of rapid pacing. In conclusion, these data suggest that autonomically mediated PV ectopy may be due to Ca2+-mediated triggered activity arising from delayed afterdepolarizations.
Role of glutamate in the rostral ventrolateral medulla in acupuncture-related modulation of visceral reflex sympathoexcitation
Authors: Zhou Wei , Fu Liang-Wu , Guo Zhi-Ling , Longhurst John C.
Published: December 2006

Visceral sympathoexcitatory reflexes induced by stimulation of the gallbladder with bradykinin (BK) are attenuated by electroacupuncture (EA) at Neiguan-Jianshi (P5-6) acupoints located over the median nerve. Previous studies have shown that neurons in the rostral ventrolateral medulla (rVLM) receive convergent input from visceral organs and somatic nerves (activated by EA). Glutamate (Glu), an important excitatory neurotransmitter in the rVLM, processes visceral sympathoexcitatory cardiovascular reflexes. In the present study, we determined the relation between EA-mediated opioidergic modulation of visceral cardiovascular responses and Glu. Reflex cardiovascular responses were evoked by application of BK to the gallbladder before and after EA in anesthetized cats. Glu concentrations ([Glu]) were measured by HPLC from samples collected by microdialysis probe(s) inserted unilaterally or bilaterally into the rVLM. BK-induced reflex responses and [Glu] were attenuated by 45% and 70%, respectively, after 30 min of EA ( n = 6). EA alone did not change [Glu] in the rVLM ( n = 6, P > 0.05). However, microdialysis of naloxone (100 mM) into the rVLM reversed EA-related inhibition of blood pressure and [Glu] ( n = 5). Immunohistochemical visualization showed that δ-opioid receptors colocalized with, and were in close apposition to, vesicular Glu transporter 3- and c-Fos-double-labeled perikarya and processes of rVLM neurons after gallbladder stimulation with BK. These data suggest that EA attenuates BK-induced visceral sympathoexcitatory reflexes through opioid-mediated inhibition of Glu's action in the rVLM.
Laser microdissection and capture of pure cardiomyocytes and fibroblasts from infarcted heart regions: perceived hyperoxia induces p21 in peri-infarct myocytes
Authors: Kuhn Donald E. , Roy Sashwati , Radtke Jared , Khanna Savita , Sen Chandan K.
Published: December 2006

Myocardial infarction caused by ischemia-reperfusion in the coronary vasculature is a focal event characterized by an infarct-core, bordering peri-infarct zone and remote noninfarct zone. Recently, we have reported the first technique, based on laser microdissection pressure catapulting (LMPC), enabling the dissection of infarction-induced biological responses in multicellular regions of the heart. Molecular mechanisms in play at the peri-infarct zone are central to myocardial healing. At the infarct site, myocytes are more sensitive to insult than robust fibroblasts. Understanding of cell-specific responses in the said zones is therefore critical. In this work, we describe the first technique to collect the myocardial tissue with a single-cell resolution. The infarcted myocardium was identified by using a truncated hematoxylin-eosin stain. Cell elements from the infarct, peri-infarct, and noninfarct zones were collected in a chaotropic RNA lysis solution with micron-level surgical precision. Isolated RNA was analyzed for quality by employing microfluidics technology and reverse transcribed to generate cDNA. Purity of the collected specimen was established by real-time PCR analyses of cell-specific genes. Previously, we have reported that the oxygen-sensitive induction of p21/Cip1/Waf1/Sdi1 in cardiac fibroblasts in the peri-infarct zone plays a vital role in myocardial remodeling. Using the novel LMPC technique developed herein, we confirmed that finding and report for the first time that the induction of p21 in the peri-infarct zone is not limited to fibroblasts but is also evident in myocytes. This work presents the first account of an analytical technique that applies the LMPC technology to study myocardial remodeling with a cell-type specific resolution.
Modified LDLs induce proliferation-mediated death of human vascular endothelial cells through MAPK pathway
Authors: Apostolov Eugene O. , Basnakian Alexei G. , Yin Xiaoyan , Ok Ercan , Shah Sudhir V.
Published: December 2006

The ability of modified low-density lipoptoteins (LDLs) to induce both proliferation and death of endothelial cells is considered to be a mechanism of early atherosclerosis development. We previously showed that carbamylated LDL (cLDL) induces human coronary artery endothelial cell (HCAEC) death in vitro. This effect is similar to the atherogenic action of oxidized LDL (oxLDL) that induces the proliferation and death of endothelial cells. The present study was designed to analyze a potential proliferative effect of cLDL and whether proliferation caused by modified LDLs is related to cell death. Cultured HCAECs were exposed to different concentrations of modified LDL or native LDL for varying periods of time. Cell proliferation measured by bromodeoxyuridine incorporation and S-phase analysis was dose-dependently increased in the presence of cLDL (6.25–200 μg/ml). The proliferation induced by cLDL or oxLDL was associated with cell death and increased phosphorylation of extracellular signal-regulated kinase (ERK) and c-Jun NH2-terminal kinase (JNK). Inhibition of cLDL- or oxLDL-induced proliferation by aphidicolin (1 μg/ml) was protective against both short-term cell death measured by lactate dehydrogenase release into the medium and long-term cell viability visualized by cell multiplication. Inhibition of ERK phosphorylation led to a significant decrease of DNA synthesis and cell rescue from injury by modified LDLs, while inhibition of JNK phosphorylation had an only partial rescue effect without involvement in cell proliferation. These data are the first evidence that endothelial cell death induced by cLDL or oxLDL is mediated by cell proliferation through the mitogen-activated protein kinase pathway.
A peptide inhibitor of c-Jun NH2-terminal kinase reduces myocardial ischemia-reperfusion injury and infarct size in vivo
Authors: Milano Giuseppina , Morel Sandrine , Bonny Christophe , Samaja Michele , von Segesser Ludwig K. , Nicod Pascal , Vassalli Giuseppe
Published: December 2006

The c-Jun NH2-terminal kinase (JNK) pathway of the mitogen-activated protein kinase (MAPK) signaling cascade regulates cell function and survival after stress stimulation. Equally robust studies reported dichotomous results suggesting both protective and detrimental effects of JNK during myocardial ischemia-reperfusion (I/R). The lack of a highly specific JNK inhibitor contributed to this controversy. We recently developed a cell-penetrating, protease-resistant peptide inhibitor of JNK, d-JNKI-1. Here we report on the effects of d-JNKI-1 in myocardial I/R. d-JNKI-1 was tested in isolated-perfused adult rat hearts. Increased activation of JNK, p38-MAPK, and extracellular signal-regulated kinase-1/2 (ERK1/2), as assessed by kinase assays and Western blotting, occurred during I/R. d-JNKI-1 delivered before onset of ischemia prevented the increase in JNK activity while not affecting ERK1/2 and p38-MAPK activation. JNK inhibition reduced ischemic injury, as manifested by increased time to contracture ( P < 0.05) and decreased left ventricular end-diastolic pressure during ischemia ( P < 0.01), and enhanced posthypoxic recovery of systolic and diastolic function ( P < 0.01). d-JNKI-1 reduced mitochondrial cytochrome- c release, caspase-3 activation, and the number of apoptotic cells determined by terminal deoxynucleotidyltransferase-mediated dUTP nick-end labeling ( P < 0.05), indicating suppression of the mitochondrial machinery of apoptosis. d-JNKI-1 delivered at the time of reperfusion did not improve functional recovery but still prevented apoptosis. In vivo, d-JNKI-1 reduced infarct size after coronary artery occlusion and reperfusion by ∼50% ( P < 0.01). In conclusion, d-JNKI-1 is an important compound that can be used in preclinical models to investigate the role of JNK signaling in vivo. Inhibition of JNK during I/R is cardioprotective in anesthetized rats in vivo.
Role of oxidative stress in angiotensin II-induced enhanced expression of Giα proteins and adenylyl cyclase signaling in A10 vascular smooth muscle cells
Authors: Li Yuan , Lappas Georgios , Anand-Srivastava Madhu B.
Published: December 2006

We have previously reported that angiotensin II (ANG II) treatment of A10 vascular smooth muscle cells (VSMCs) increased inhibitory G proteins (Giprotein) expression and associated adenylyl cyclase signaling which was attributed to the enhanced MAP kinase activity. Since ANG II has been shown to increase oxidative stress, we investigated the role of oxidative stress in ANG II-induced enhanced expression of Giα proteins and examined the effects of antioxidants on ANG II-induced enhanced expression of Giα proteins and associated adenylyl cyclase signaling in A10 VSMCs. ANG II treatment of A10 VSMCs enhanced the production of O2−and the expression of Nox4 and P47phox, different subunits of NADPH oxidase, which were attenuated toward control levels by diphenyleneiodonium (DPI). In addition, ANG II augmented the expression of Giα-2 and Giα-3 proteins in a concentration- and time-dependent manner; the maximal increase in the expression of Giα was observed at 1 to 2 h and at 0.1–1.0 μM. The enhanced expression of Giα-2 and Giα-3 proteins was restored to control levels by antioxidants such as N-acetyl-l-cysteine, α-tocopherol, DPI, and apocynin. In addition, ANG II also enhanced the ERK1/2 phosphorylation that was restored to control levels by DPI. Furthermore, the inhibition of forskolin-stimulated adenylyl cyclase activity by low concentrations of 5′- O-(3-triotriphosphate) (receptor-independent Gifunctions) and ANG II-, des(Glu18,Ser19,Glu20,Leu21,Gly22)atrial natriuretic peptide4-23-NH2(natriuretic peptide receptor-C agonist), and oxotremorine-mediated inhibitions of adenylyl cyclase (receptor-dependent functions) that were augmented in ANG II-treated VSMCs was also restored to control levels by antioxidant treatments. In addition, Gsα-mediated diminished stimulation of adenylyl cyclase by stimulatory hormones in ANG II-treated cells was also restored to control levels by DPI. These results suggest that ANG II-induced enhanced levels of Giα proteins and associated functions in VSMCs may be attributed to the ANG II-induced enhanced oxidative stress, which exerts its effects through mitogen-activated protein kinase signaling pathway.
Paradoxical effect of salbutamol in a model of acute organophosphates intoxication in guinea pigs: role of substance P release
Authors: Chávez Jaime , Segura Patricia , Vargas Mario H. , Arreola José Luis , Flores-Soto Edgar , Montaño Luis M.
Published: December 2006

Organophosphates induce bronchoobstruction in guinea pigs, and salbutamol only transiently reverses this effect, suggesting that it triggers additional obstructive mechanisms. To further explore this phenomenon, in vivo (barometric plethysmography) and in vitro (organ baths, including ACh and substance P concentration measurement by HPLC and immunoassay, respectively; intracellular Ca2+measurement in single myocytes) experiments were performed. In in vivo experiments, parathion caused a progressive bronchoobstruction until a plateau was reached. Administration of salbutamol during this plateau decreased bronchoobstruction up to 22% in the first 5 min, but thereafter airway obstruction rose again as to reach the same intensity as before salbutamol. Aminophylline caused a sustained decrement (71%) of the parathion-induced bronchoobstruction. In in vitro studies, paraoxon produced a sustained contraction of tracheal rings, which was fully blocked by atropine but not by TTX, ω-conotoxin (CTX), or epithelium removal. During the paraoxon-induced contraction, salbutamol caused a temporary relaxation of ∼50%, followed by a partial recontraction. This paradoxical recontraction was avoided by the M2- or neurokinin-1 (NK1)-receptor antagonists (methoctramine or AF-DX 116, and L-732138, respectively), accompanied by a long-lasting relaxation. Forskolin caused full relaxation of the paraoxon response. Substance P and, to a lesser extent, ACh released from tracheal rings during 60-min incubation with paraoxon or physostigmine, respectively, were significantly increased when salbutamol was administered in the second half of this period. In myocytes, paraoxon did not produce any change in the intracellular Ca2+basal levels. Our results suggested that: 1) organophosphates caused smooth muscle contraction by accumulation of ACh released through a TTX- and CTX-resistant mechanism; 2) during such contraction, salbutamol relaxation is functionally antagonized by the stimulation of M2receptors; and 3) after this transient salbutamol-induced relaxation, a paradoxical contraction ensues due to the subsequent release of substance P.
TNF-α induces MMP-9 expression via activation of Src/EGFR, PDGFR/PI3K/Akt cascade and promotion of NF-κB/p300 binding in human tracheal smooth muscle cells
Authors: Lee Chiang-Wen , Lin Chih-Chung , Lin Wei-Ning , Liang Kao-Chih , Luo Shue-Feng , Wu Chow-Bin , Wang Shyi-Wu , Yang Chuen-Mao
Published: December 2006

TNF-α has been shown to induce matrix metalloproteinase-9 (MMP-9) expression, which, in turn, degrades extracellular matrix in the inflammatory responses. However, the inductive mechanisms of the MMP-9 by TNF-α remain unclear. In human tracheal smooth muscle cells, TNF-α induced MMP-9 expression and Akt phosphorylation in a time-dependent manner, which was attenuated by the inhibitors of Src (PP1), epidermal growth factor receptor (AG1478), PDGFR (AG1296), and PI3K (LY294002), respectively, revealed by reporter gene assay, RT-PCR, zymographic, and Western blot analyses. Transfection with the dominant negative mutants of c-Src (KM, K295M [kinase inactive mutant]), p85, and Akt (KA, K179A) also reduced MMP-9 expression. These findings indicated that MMP-9 expression was regulated by PI3K/Akt via the transactivation of growth factor receptors. Furthermore, LY294002 or wortmannin inhibited Akt phosphorylation but had no effect on NF-κB translocation, which was blocked by helenalin. Mutated NF-κB DNA binding element in the MMP-9 promoter and helenalin also attenuated MMP-9 expression, suggesting that PI3K/Akt and NF-κB independently regulated MMP-9 expression. To support this notion, immunofluorescence staining and immunoprecipitation were applied to characterize the transcription factors involved in these responses. The results showed that LY294002 and curcumin blocked Akt translocation into nucleus. In contrast, p300, acetyl-histone (H3), and NF-κB p65 were found to be coimmunoprecipitated with the phosphorylated Akt, indicating that these components associated with the MMP-9 promoter are revealed by chromatin immunoprecipitation assay. Thus, our study provides a new insight into the molecular mechanisms that TNF-α-stimulated Akt phosphorylation mediated through transactivation of Src and growth factor receptors may stimulate the recruitment of p300, assemble transcription factor (p65), and then lead to MMP-9 expression.
Elevated amount of Toll-like receptor 4 mRNA in bronchial epithelial cells is associated with airway inflammation in horses with recurrent airway obstruction
Authors: Berndt Annerose , Derksen Frederik J. , Venta Patrick J. , Ewart Susan , Yuzbasiyan-Gurkan Vilma , Robinson N. Edward
Published: December 2006

Recurrent airway obstruction (RAO) is characterized by neutrophilic airway inflammation and obstruction, and stabling of susceptible horses triggers acute disease exacerbations. Stable dust is rich in endotoxin, which is recognized by Toll-like receptor (TLR) 4. In human bronchial epithelium, TLR4 stimulation leads to elevation of interleukin (IL)-8 mRNA expression. The zinc finger protein A20 negatively regulates this pathway. We hypothesized that TLR4 and IL-8 mRNA and neutrophil numbers are elevated and that A20 mRNA is not increased in RAOs during stabling compared with controls and with RAOs on pasture. We measured the maximal change in pleural pressure (ΔPplmax), determined inflammatory cell counts in bronchoalveolar lavage fluid (BAL), and quantified TLR4, IL-8, and A20 mRNA in bronchial epithelium by quantitative RT-PCR. We studied six horse pairs, each pair consisting of one RAO and one control horse. Each pair was studied when the RAO-affected horse had airway obstruction induced by stabling and after 7, 14, and 28 days on pasture. Stabling increased BAL neutrophils, ΔPplmax, and TLR4 (4.14-fold change) significantly in RAOs compared with controls and with RAOs on pasture. TLR4 correlated with IL-8 ( R2= 0.75). Whereas stabling increased IL-8 in all horses, A20 was unaffected. IL-8 was positively correlated with BAL neutrophils ( R2= 0.43) and negatively with A20 ( R2= 0.44) only in RAO-affected horses. Elevated TLR4 expression and lack of A20 upregulation in bronchial epithelial cells from RAO-affected horses may contribute to elevated IL-8 production, leading to exaggerated neutrophilic airway inflammation in response to inhalation of stable dust.
Effect of hydrogen peroxide on ROMK channels in the cortical collecting duct
Authors: Wei Yuan , Wang ZhiJian , Babilonia Elisa , Sterling Hyacinth , Sun Peng , Wang WenHui
Published: December 2006

We used the patch-clamp technique to study the effect of H2O2 on the apical ROMK-like small-conductance K (SK) channel in the cortical collecting duct (CCD). The addition of H2O2 decreased the activity of the SK channels and the inhibitory effect of H2O2 was larger in the CCD from rats on a K-deficient diet than that from rats on a normal-K or a high-K diet. However, application of H2O2 did not inhibit the SK channels in inside-out patches. This suggests that the H2O2-mediated inhibition of SK channels was not due to direct oxidation of the SK channel protein. Because a previous study showed that H2O2 stimulated the expression of Src family protein tyrosine kinase (PTK) which inhibited SK channels ( 3 ), we explored the role of PTK in mediating the effect of H2O2 on SK channels. The application of H2O2 stimulated the activity of endogenous PTK in M-1 cells and increased tyrosine phosphorylation of ROMK in HEK293 cells transfected with GFP-ROMK1 and c-Src. However, blockade of PTK only attenuated but did not completely abolish the inhibitory effect of H2O2 on SK channels. Since H2O2 has also been demonstrated to activate mitogen-activated protein kinase, P38, and ERK ( 3 ), we examined the role of P38 and ERK in mediating the effect of H2O2 on SK channels. Similar to blockade of PTK, suppression of P38 and ERK did not completely abolish the H2O2-induced inhibition of SK channels. However, combined use of ERK, P38, and PTK inhibitors completely abolished the effect of H2O2 on SK channels. Also, treatment of the CCDs with concanavalin A, an agent which has been shown to inhibit endocytosis ( 19 ), abolished the inhibitory effect of H2O2. We conclude that addition of H2O2 inhibited SK channels by stimulating PTK activity, P38, and ERK in the CCD and that H2O2 enhances the internalization of the SK channels.
Use of calibrated automated thrombinography ± thrombomodulin to recognise the prothrombotic phenotype
Authors: Trzeciak M. , Bordet Jean , Ninet Jacques , Negrier Claude , Dargaud Yesim
Published: December 2006

There is currently no validated method to detect a prothrombotic phenotype. The question remains, can tissue factor (TF) induced thrombin generation (TG), as measured with the calibrated automated thrombinography (CAT) technique, according to Hemker et al., recognise a prothrombotic state either as such, or when the activated proteinC (APC)-system is boosted with thrombomodulin (TM)? We determined the normal range of CAT-TG ± TM in a group of 71 healthy blood donors, in 11 healthy women using oral contraceptives (OC), and in 89 patients with a history of venous thromboembolism (VTE), divided into a group of 50 in whicha prothrombotic risk factor could be found (VTEprf+) and 39 others (VTEprf-). The endogenous thrombin potential (ETP) in the OC, VTEprf+ and VTEprf-group was significantly higher than for the controls. In the presence of TM, the differences were significantly higher than in its absence. The VTEprf+ group had a higher ETP, ± TM than the VTEprf group. In conclusion,TG, measured with the CAT technique in the presence of TM is capable of detecting the prothrombotic phenotype with a high sensitivity of 0.93 (95% confidence limits 0.82–0.99).
Management of cutaneous type IV hypersensitivity reactions induced by heparin
Authors: Schindewolf Marc , Utikal Jochen , Lindhoff-Last Edelgard , Boehncke Wolf-Henning , Ludwig Ralf
Published: December 2006

Localized hypersensitivity reactions to subcutaneous heparin injections have been described since 1952. Yet, the incidence of these reactions, which are distinct from skin lesions associated with heparin-induced thrombocytopenia type II (HIT II), remains uncertain. However, in the last 10 years an increasing number of patients have been reported, leading to the assumption that cutaneous hypersensitivity reactions towards heparin are underreported. Clinically patients present with itching, sometimes infiltrated, and blistering erythemas at the injection sites of heparins. The diagnosis of cutaneous heparin allergy may, on the one hand, lead to delay of required medical or surgical treatment. On the other hand, delayed initiation of treatment may lead to a generalized eczematous reaction. Hence, from review of 223 cases of patients with cutaneous hypersensitivity reactions to heparin, we here summarize the clinical picture of cutaneous type IV allergic reactions, define risk factors on both the patient- and drug-side, and give an overview of principle therapeutic alternatives, as well as recommendations for treatment options for emergency and elective patients. As the proposed management of patients with cutaneous hypersensitivity reactions to heparin may have fatal consequences when applied in patients with HIT type II, diagnosis of skin lesions in heparin-treated patients needs to be precise.
SELDI-TOF plasma profiles distinguish individuals in a protein C-deficient family with thrombotic episodes occuring before age 40
Authors: Svensson Annika , Whiteley Gordon , Callas Peter , Bovill Edwin
Published: December 2006

We tested the hypothesis that differences in the low-molecularweight (500–20,000 Da) proteomic profile of plasma may be detectable between members ofa protein C-deficient family who have suffered thrombotic events before age 40 compared to family members without a history of venous thrombosis. Unfractionated plasma samples from members of a previously described large thrombophilic kindred with type I protein C deficiency were applied to ProteinChip weak cation exchange interaction arrays (WCX2; Ciphergen Biosystems, Fremont, CA, USA) and subjected to SELDI-TOF (surface-enhanced laser desorption/ionization time-of-flight) mass spectrometry using the Ciphergen PBSII ProteinChip System (Ciphergen Biosystems). Profiles were analyzed by a boosted decision-tree algorithm. When individuals who had presented with deep venous thrombosis (DVT) before the age of 40 (n=21) were compared to age-matched, healthy family members (n=50), the proteomic patterns defined by the decision-tree analysis could classify the entity of DVT before age 40 with 67% sensitivity, ata specificity of 86%.Whena small group of cases with history of superficial venous thrombosis (n=6) was added to the case group, the sensitivity was 87.5% at a specificity of 80%.These data support the hypothesis that members of the protein C deficient Vermont kindred II who suffer a thrombotic event before age 40 display significant differences in low-molecular-weight proteomics profile compared to those who remain disease-free.This is the first study to apply SELDI-TOF technology in conjunction with a bioinformatics tool to analyze low-molecular-weight proteomic patterns in patients with venous thrombosis.

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Anti-metastatic effect of a non-anticoagulant low-molecular-weight heparin versus the standard low-molecular-weight heparin, enoxaparin
Authors: Linhardt Robert , Francis John , Amirkhosravi Ali , Mousa Shaker
Published: December 2006

Low-molecular-weight heparins (LMWH) exhibit potent anticoagulant efficacy via their plasmatic effects on thrombin and factor Xa. These agents are also effective in releasing endothelial tissue factor pathway inhibitor (TFPI),the natural inhibitor of tissue factor, and exhibit significant anti-metastatic effects in experimental animal models. However, the potential for bleeding complications has slowed down the more widespread adoption of LMWH therapy in cancer patients. In this study, the effect of a non-anticoagulant form of LMWH (NA-LMWH) on experimental lung metastasis and tumor cell-induced platelet aggregation in vivo was compared to the LMWH enoxaparin. Using the B16 melanoma mouse model of metastasis, subcutaneous (s.c.) injection of NA-LMWH or enoxaparin (10 mg/kg), three hours before intravenous (i.v.) injection of metastatic melanoma cells, followed by daily doses for 14 days, reduced lung tumor formation by 70% (P<0.001). I.v. injection of tumor cells resulted in a significant (50–62%, P<0.01) fall in platelet counts. Pre-injection (i.v.) of enoxaparin completely abolished the tumor cell-induced thrombocytopenia, whereas NA-LMWH had no effect. Four hours after a single s.c. dose, enoxaparin but not NALMWH prolonged the clotting time three-fold and delayed the time to clot initiation more than 10-fold as measured by a Sonoclot analyzer and by thromboelastography, respectively. Enoxaparin but not NA-LMWH demonstrated a significant anticoagulant effect in mice. Both NA-LMWH and enoxaparin caused similar TFPI release from endothelial cells in vitro.These data provide evidence to support the potential of NA-LMWH as an anti-metastatic agent without any significant impact on coagulation.