bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cutaneous CD30+ Cells in Children with Atopic Dermatitis
Authors: Cavagni Giovanni , Caffarelli Carlo , Facchetti Fabio , Brugnoni Duilio , Notarangelo Luigi D. , Tosoni Cinzia , Altobelli Roberto 
Published: April 2003

<i>Background:</i> CD30 expression can be considered a marker of Th2 cells. We investigated the presence of CD30+ cells in the lesional skin of children with atopic dermatitis (AD). We also analyzed the possible relationship between CD30+ cells and serum soluble CD 30 (sCD30) levels, and IgE, soluble interleukin-2 (IL-2) receptor (sIL-2R) or soluble CD23 (sCD23) levels in the blood, and clinical score. <i>Methods:</i> Ten eczematous children (4 males, 6 females; median age: 4 years and 5 months; range: 11 months to 14 years), 9 sex- and age-matched control children and an adult control group were studied. A clinical score (SCORAD index), was given to eczematous lesions. Blood was taken for the determination of IgE, sCD30, sIL-2R and sCD23 levels. Punch biopsies of lesional skin were stained with hematoxylin and eosin or incubated with anti-CD30 monoclonal antibodies. Skin prick tests (SPTs) were also performed. <i>Results:</i> In the biopsy specimens, CD30 expression was observed in high proportions of infiltrating cells. In children with AD, total serum IgE, sCD30, sIL-2R, sCD23 and eosinophils were significantly elevated compared to controls. CD30+ cells were not associated with serum IgE, sCD30, sIL-2R, sCD23, or SPT results, score of inflammatory cells in lesional skin or clinical score. Children with AD who had high total IgE and specific IgE antibodies did not differ from those with normal total IgE and negative specific IgE in respect of age, clinical score, number of CD30+ cells, sCD30, sIL-2R and sCD23 levels, score of inflammatory cells in skin or clinical score. <i>Conclusion:</i> Our results showed remarkable numbers of CD30+ cells in the lesional skin and high sCD30 in the serum of children with AD. CD30+ cells did not correlate with systemic markers of IgE reaction.
Development of a Monoclonal Antibody-Based Sandwich ELISA for Detection of the Latex Allergen Hev b 1
Authors: Raulf-Heimsoth Monika , Sander Ingrid , Chen Zhiping , Borowitzki Gerda , Diewald Kai , van Kampen Vera , Baur Xaver 
Published: April 2003

<i>Background:</i> Natural rubber latex (NRL) products are complex mixtures consisting of different allergenic components. Among them, Hev b 1 belongs to the important and well-characterized ones. To quantify the relevant allergen Hev b 1 in NRL products, a two-site monoclonal antibody (mAb)-based assay was developed. <i>Methods:</i> Two Hev b 1-specific mAbs with different epitope recognition and ability to bind simultaneously to an Hev b 1 molecule were used in the study. Both mAbs (II4F9 and II4G9) were enriched by in vitro production in a modular minifermenter and affinity purified. Wells of micro-ELISA plates coated with captured mAb II4G9 were incubated with samples containing Hev b 1. Bound Hev b 1 was detected by a combination of biotinylated mAb II4F9 as detection antibody and peroxidase-labeled avidin. <i>Results:</i> The optimized sandwich ELISA was highly reproducible in the linear range of the standard curve and Hev b 1 concentrations ranging from 12.5 to 400 ng/100 μl could be detected. The assay was suitable for the detection of Hev b 1 concentrations in latex sap and latex products, e.g. gloves, with a detection limit of 1.25 μg of Hev b 1/g of rubber. In a preliminary study with five different brands of latex gloves, Hev b 1 concentrations were found to be in the range of 18–40 μg per gram of rubber material, corresponding to 2–4% of the total extractable protein content in latex glove extracts. <i>Conclusions:</i> A sensitive sandwich assay was developed to quantify the latex allergen Hev b 1. This assay can be used to standardize latex extracts with regard to the content of the major allergen Hev b 1.
Effects of EGF and TGF-α on Invasion and Proteinase Expression of Uterine Cervical Adenocarcinoma OMC-4 Cells
Authors: Ueda Masatsugu , Fujii Hideji , Yoshizawa Keiko , Terai Yoshito , Kumagai Koji , Ueki Ken , Ueki Minoru 
Published: April 2003

Uterine cervical adenocarcinoma typically is an aggressive neoplasm with a propensity for early invasion and dissemination; however, the regulatory mechanism of invasive activity of cervical adenocarcinoma cells has not been fully understood. In this study, biological effects of epidermal growth factor (EGF) and transforming growth factor (TGF)-α on invasion and proteinase expression of human cervical adenocarcinoma OMC-4 cells were investigated. Tumor cell migration along a gradient of substratum-bound fibronectin and invasion into the reconstituted basement membrane were stimulated by 0.1–10 n<i>M</i> EGF and TGF-α in a concentration-dependent manner. Their effects on tumor cell migration were also confirmed by wound assay. The zymography of tumor-conditioned medium showed that the treatment of OMC-4 cells with EGF and TGF-α resulted in the increase of matrix metalloproteinase (MMP)-2 and urokinase-type plasminogen activator (uPA). Matrilysin (MMP-7), also secreted by OMC-4 cells, was not affected by these growth factors. These results suggest that EGF and TGF-α act as positive regulators on the invasion of cervical adenocarcinoma cells, which may be associated with their stimulatory effects on tumor cell motility and the induction of type IV collagenase and uPA secreted by tumor cells.
The Late Expression Factors 8 and 9 and Possibly the Phosphoprotein p78/83 of <i>Autographa californica</i> Multicapsid Nucleopolyhedrovirus Are Components of the Virus-Induced RNA Polymerase
Authors: Iorio Caterina , Vialard Jorge E. , McCracken Susan , Lagacé Monique , Richardson Christopher D. 
Published: April 2003

Attempts were made to identify some of the subunits of the baculovirus-induced RNA polymerase following purification of its enzymatic activity by conventional chromatography. Polymerase activity was extracted from lysates of insect cells infected with <i>Autographa californica</i> multicapsid nucleopolyhedrovirus by polyethylenimine precipitation and subsequently purified by phosphocellulose, anion exchange, poly(A) Sepharose affinity, and gel filtration chromatography. The presence of the polymerase was monitored by its α-amanitin-resistant activity in in vitro transcription assays. A number of polypeptides associated with the enzymatic activity were identified. Peptide-specific antibodies were generated against a variety of late-expression factors (LEFs) and these antibodies, along with antisera directed against several other baculovirus proteins, were used in an immunoblot analysis of the purified polymerase. The results revealed that both the viral helicase (p143) and the virogenic stroma protein, pp31, copurify with the baculovirus-induced RNA polymerase activity through several chromatographic steps and may be loosely associated with the RNA polymerase. LEF8, LEF9 and p78/83, a nucleocapsid-associated phosphoprotein, were found to associate with the viral-induced polymerase activity. LEF8 and LEF9 contain regions of sequence homology with components of other DNA-directed RNA polymerases, while a portion of p78/83 exhibits some homology to the sigma factor of bacterial RNA polymerase, the RAP30 protein found in the mammalian transcription complex TFIIF, and the RAP94 polypeptide associated with vaccinia virus RNA polymerase. The p78/83 protein has previously been shown by our laboratory to be a capsid protein, but it may also play some role with the RNA polymerase. These results represent a first attempt to identify specific components of the RNA polymerase associated with infections of insect cells by <i>A. californica</i> nucleopolyhedrovirus.
Wall Remodeling after Wall Shear Rate Normalization in Rat Mesenteric Arterial Collaterals
Authors: Fath Steven W. , Burkhart Harold M. , Miller Scott C. , Dalsing Michael C. , Unthank Joseph L. 
Published: April 2003

Previous studies have demonstrated endothelial and smooth muscle hyperplasia occur during arterial luminal expansion associated with elevation of arterial wall shear rates. The current study investigated whether remodeling induced by elevated wall shear would ultimately result in a vessel with intimal and medial cell densities and other wall characteristics similar to control arteries. A rat mesenteric model was used in which collateral wall shear is restored to normal 4 weeks after arterial occlusion. Twelve weeks after shear elevation, paired in vivo measurements indicated that the maximum collateral inner diameter was increased 27–75%. Morphometric evaluation of collateral cross sections indicated that, relative to control arteries, luminal and medial areas were increased 79 ± 22 and 56 ± 15%. Collateral medial cell density was decreased (1.12 ± 0.044 vs. 1.35 ± 0.005 nuclei/1,000 µm<sup>2</sup>) but intimal cell density was similar (2.86 ± 0.166 vs. 2.49 ± 0.102 nuclei/100 µm luminal perimeter). Medial thickness to radius ratio was also similar between control and collateral arteries. Thus, for the wall characteristics evaluated, there are many similarities between enlarged collaterals and control arteries. Comparison of nuclear numbers in arterial cross sections of the current and previous studies suggest that intimal and medial cellular regression is correlated with a decrease in wall shear force toward normal levels.
Characterization of Endothelium- Dependent Relaxation in Guinea Pig Basilar Artery – Effect of Hypoxia and Role of Cytochrome P<sub>450</sub> Mono-Oxygenase
Authors: Petersson Jesper , Zygmunt Peter M. , Jönsson Peter , Högestätt Edward D. 
Published: April 2003

In the guinea pig basilar artery, acetylcholine and the calcium ionophore A23187 induced endothelium-dependent relaxations, which were not significantly affected by the nitric oxide (NO) synthase inhibitor N<sup>ω</sup>-nitro-<i>L</i>-arginine (<i>L</i>-NOARG; 0.3 m<i>M</i>) or the guanylate cyclase inhibitor ODQ (1H-[1,2,4]oxadiazolo[4,3-a]quinoxaline-1-one; 1-10 µ<i>M</i>), or by these inhibitors combined. However, acetylcholine (10 µ<i>M</i>) and A23187 (3 µ<i>M</i>) each significantly increased the tissue level of cGMP in the absence but not in the presence of <i>L</i>-NOARG, suggesting that NO is released from the vascular endothelium in this blood vessel. Treatment with the potassium (K) channel inhibitors charybdotoxin (0.1 µ<i>M</i>) plus apamin (0.1 µ<i>M</i>), a toxin mixture previously shown to inhibit relaxations mediated by endothelium-derived hyperpolarizing factor (EDHF) in this artery, had no effect on the A23187-induced relaxation but slightly inhibited the response to acetylcholine (E<sub>max</sub> was reduced by 24%). When the action of EDHF was prevented by these K channel inhibitors, the remaining relaxation was abolished by either ODQ (1 µ<i>M</i>) or <i>L</i>-NOARG (0.3 m<i>M</i>), indicating that NO, apart from EDHF, contributes to the endothelium-dependent relaxations. Furthermore, ODQ (10 µ<i>M</i>) abolished the relaxation induced by the NO donor S-nitroso-N-acetylpenicillamine. Thus, activation of soluble guanylate cyclase seems to be the only mechanism through which NO causes relaxation in this artery. When vessels were exposed to grave hypoxia (pO<sub>2</sub> = 6 mm Hg), the NO-mediated relaxation (induced by acetylcholine in the presence of charybdotoxin plus apamin) disappeared. In contrast, EDHF-mediated responses (elicited by acetylcholine in the presence of <i>L</i>-NOARG) were only marginally affected by hypoxia (E<sub>max</sub> was reduced by 16%). 17-Octadecynoic acid (50 µ<i>M</i>) and 5,8,11,14-eicosatetraynoic acid (10 µ<i>M</i>), inhibitors of cytochrome P<sub>450</sub>-dependent oxidation of arachidonic acid, failed to inhibit the acetylcholine-induced relaxation in the presence of <i>L</i>-NOARG. The cytochrome P<sub>450</sub>-dependent arachidonic acid metabolite 11,12-epoxyecosatrienoic acid (0.3–3.0 µ<i>M</i>) had no relaxant effect per se. In conclusion, EDHF and NO are both mediators of endothelium-dependent relaxations in the guinea pig basilar artery. However, during grave hypoxia, EDHF alone mediates acetylcholine-induced relaxation. The results further suggest that EDHF is not a metabolite of arachidonic acid formed by cytochrome P<sub>450</sub> mono-oxygenase or generated by another oxygen-dependent enzyme in this artery.
Medial and Endothelial Platelet-Derived Growth Factor A Chain Expression Is Regulated by in vivo Exposure to Elevated Flow
Authors: Tulis David A. , Prewitt Russell L. 
Published: April 2003

Previous work from this laboratory demonstrated that in vivo exposure to elevated arterial flow stimulates endothelial and smooth muscle cell hyperplasia concomitant with lumen enlargement, medial wall hypertrophy, and increases in medial extracellular connective tissue in rat mesenteric small arteries. In an effort to elucidate the role of growth factors in mediating this arterial remodeling response, in situ hybridization was performed on control and high flow arterial sections using <sup>35</sup>S-labeled riboprobes for PDGF-A, PDGF-B, basic FGF, and TGFβ<sub>1</sub> mRNA. Results demonstrate that after exposure to elevated arterial flow for 24 h, expression of PDGF-A mRNA in the media was significantly elevated over basal levels (+215%, p < 0.001). This expression decreased towards control levels by 3 and 7 days. Increased endothelial expression of PDGF-A mRNA over basal levels was evident after 3 and 7 days of elevated flow (+129%, p < 0.01; +182%; p < 0.01, respectively). Acute medial PDGF-A mRNA expression may result from elevated circumferential hoop stress secondary to flow-induced dilation while delayed expression in the endothelium may result from normalization of wall shear stress. Endothelial expression of PDGF-B mRNA was significantly elevated over control levels (+62%, p < 0.01) after exposure to high flow for 7 days. Expression of bFGF and TGFβ<sub>1</sub> mRNA was not significantly different between control and high flow vessels at all times measured. These results suggest a role for PDGF-A in regulating acute arterial remodeling following in vivo exposure to elevated flow.
Endothelin-1 Stimulates Hydrolysis of Phosphatidylcholine by Phospholipases C and D in Intact Rat Mesenteric Arteries
Authors: Liu GeLe , Shaw Linda , Heagerty AnthonyM. , Ohanian Vasken , Ohanian Jacqueline 
Published: April 2003

A characteristic of endothelin-1 (ET-1)-induced contraction is the prolonged duration of the response. Phosphatidylcholine (PC) hydrolysis has been implicated in sustained agonist-induced effects through the formation of the second messengers phosphatidic acid (PA) and diacylglycerol (DAG). Therefore, we have investigated the activation of PC-phospholipase D (PLD) and PC-phospholipase C (PLC) in intact rat mesenteric small arteries stimulated with ET-1 and determined whether PC-derived DAG is necessary for ET-1-induced contraction. PLD activity, measured as phosphatidylethanol (PEt) production in vessels labelled with [<sup>3</sup>H] or [<sup>14</sup>C]myristate, was increased in a concentration- and time-dependent manner following ET-1 stimulation, as were [<sup>3</sup>H]PA levels, peaking at 10 min (ET-1 100 nM) and remaining above control levels for up to 20 min. Inclusion of 0.5% ethanol during ET-1 stimulation reduced [<sup>3</sup>H]PA levels, but did not alter the time course of formation. In addition, [<sup>14</sup>C]choline release was increased, confirming PLD-mediated PC hydrolysis. In contrast, DAG levels, measured by [<sup>3</sup>H]myristate labelling and mass assay, increased transiently at 5 min of ET-1 stimulation only. Analysis of the subclasses of DAG demonstrated an increase in all types of DAG without any enrichment with arachidonate-containing species, indicating PC, not inositol lipid hydrolysis was the source of DAG. The PC-PLC inhibitor D609, 2.5 µg/ml, completely abolished the ET-1-induced increase in [<sup>14</sup>C]DAG without affecting the increase in [<sup>14</sup>C]PA, PLD activity or the contractile response. In [<sup>14</sup>C]choline-labelled vessels, [<sup>14</sup>C]phosphocholine levels were increased by ET-1 with a similar time course to DAG production. Removal of extracellular Ca<sup>2+</sup> and addition of 0.1 or 2 mM EGTA completely inhibited ET-1-stimulated PLD activity. The tyrosine kinase inhibitor tyrphostin A23 (100 µM) abolished ET-1-induced [<sup>3</sup>H]PA, [<sup>3</sup>H]PEt and [<sup>14</sup>C]DAG increases, whilst the negative analogue A1 was without effect. These data suggest that ET-1 couples via a calcium-dependent tyrosine kinase mechanism to PLD and PC-PLC in vascular smooth muscle. PC-derived DAG did not appear to be necessary for the contractile response, whereas sustained formation of PA, generated by PLD activity, implies that this lipid second messenger could be involved in the prolonged contractile response to this peptide.
High-Dose, Not Low-Dose Insulin Increases the Vasoconstrictor Effect of Norepinephrine in Spontaneously Hypertensive Rats: Effects of Antihypertensive Treatment
Authors: Rizzoni Damiano , Porteri Enzo , Piccoli Alfonso , Castellano Maurizio , Pasini Giancarlo , Guelfi Daniele , Muiesan Maria Lorenza , Agabiti Rosei Enrico 
Published: April 2003

The effect of insulin on the vasoconstriction induced by norepinephrine is presently controversial. Therefore, the aims of our study were: (1) to evaluate the effect of low- and high-dose insulin on the concentration-response curve to norepinephrine in small resistance arteries of spontaneously hypertensive rats (SHR) and Wistar Kyoto rats (WKY) before and after the development of hypertension, and (2) to evaluate the effects of antihypertensive treatment on vascular response to insulin and norepinephrine. Fifty-six rats were included in the study. Six SHR were treated with enalapril and 6 with candesartan cilexetil from the 4th to the 12th week of age, while 10 WKY and 14 SHR were kept untreated. Two additional groups of 10 untreated SHR and 10 WKY were killed at 4 weeks of age, in a prehypertensive phase. Mesenteric small arteries were dissected and mounted on a micromyograph. A dose-response curve to norepinephrine was performed at cumulative concentrations in the presence or absence of low- and high-dose insulin. We found that only high-dose insulin increased the vascular response to norepinephrine in 12-week-old SHR, but not in 4-week-old SHR or in age-matched WKY. The increased responsiveness to norepinephrine disappeared after preincubation of the vessels with a selective inhibitor of endothelin-1 type A receptors. After antihypertensive treatment with enalapril or candesartan cilexetil, the potentiation of the vasoconstrictor response to norepinephrine was abolished. In conclusion, insulin at high, nonphysiological doses seems to induce an increase in the reactivity to norepinephrine in mesenteric small arteries of SHR, possibly mediated by a local production of endothelin-1. Antihypertensive treatment with an ACE inhibitor or an angiotensin II receptor blocker may normalize this altered response. This mechanism may be relevant in the development of hypertension in SHR.
Role of Voltage-Dependent and Ca<sup>2+</sup>-Activated K<sup>+</sup> Channels on the Regulation of Isometric Force in Porcine Coronary Artery
Authors: Shimizu Shunichi , Yokoshiki Hisashi , Sperelakis Nicholas , Paul Richard J. 
Published: April 2003

We investigated the role of K<sup>+</sup> channels in the regulation of vascular tone in de-endothelialized porcine coronary artery. Isometric force and intracellular Ca<sup>2+</sup> ([Ca<sup>2+</sup>]<sub>i</sub>) under resting conditions were increased by treatment with 4-aminopyridine (4-AP, 1 m<i>M</i>), an inhibitor of voltage-dependent K<sup>+</sup> (K<sub>v</sub>) channels, but not by tetraethylammonium chloride (TEA, 1 m<i>M</i>) or charybdotoxin (100 n<i>M</i>), both inhibitors of Ca<sup>2+</sup>-activated K<sup>+</sup> (K<sub>Ca</sub>) channels, or glibenclamide (10 μ<i>M</i>), an inhibitor of ATP-sensitive K<sup>+</sup> channels. Under stimulated conditions with 9,11-dideoxy-11α,9α-epoxymethano-prostaglandin F<sub>2α</sub> (U46619), 4-AP as well as TEA or charybdotoxin increased isometric force and [Ca<sup>2+</sup>]<sub>i</sub>, but not glibenclamide. 4-AP was the most potent in terms of depolarization of membrane potential compared with TEA or glibenclamide in the presence or absence of EGTA. In the presence of U46619, a high concentration of 4-AP (10 m<i>M</i>) caused a further contraction with oscillations. The force oscillations induced by 4-AP were inhibited by diltiazem (10 μ<i>M</i>), an inhibitor of voltage-dependent Ca<sup>2+</sup> channels, or TEA (1 m<i>M</i>), but not by glibenclamide (10 μ<i>M</i>). These force oscillations may be associated with the periodic activation of K<sub>Ca</sub> channels. These findings suggested that 4-AP-sensitive K<sub>v</sub> channels play an important role in the control of vascular tone in both resting and stimulated conditions. Moreover, under stimulated conditions, K<sub>Ca</sub> channels also have an important role in the regulation of vascular tone. Dysfunction of these channels induces abnormal vasoconstriction and may be implicated in vascular diseases such as hypertension and vasospasm.
Reactive Oxygen Species and Acute Modulation of Albumin Microvascular Leakage in the Microcirculation of Diabetic Rats in vivo
Authors: Bonnardel-Phu E. , Vicaut E. 
Published: April 2003

Endothelial cells have been reported to generate reactive oxygen species such as the superoxide anion, hydrogen peroxide, and the hydroxyl radical. The aim of this work was to evaluate the role of reactive oxygen species in diabetes-induced changes in vascular permeability. Intravital videomicroscopy was used to study albumin microvascular leakage in the cremaster muscle. The extravasation of a fluorescent macromolecular tracer (FITC-albumin) was measured for 1 h and, after computer-aided image analysis, was expressed as variations of normalized gray levels (arbitrary units). The extravasation of the macromolecular tracer was greater in diabetic rats (5.28 ± 1.29 vs. 1.96 ± 0.41 AU at 1 h in diabetic and control rats, respectively). Administration of superoxide dismutase (SOD), which dismutates ·O<sup>–</sup><sub>2</sub> to H<sub>2</sub>O<sub>2</sub>, and of catalase which reacts with H<sub>2</sub>O<sub>2</sub> to form H<sub>2</sub>O and molecular oxygen failed to inhibit the increased extravasation of the macromolecular tracer when administered separately. However, a significant inhibition of diabetic increase in albumin extravasation was found when these 2 drugs were administered simultaneously, and in this case, the extravasation of the macromolecular tracer at 1 h was similar in diabetic rats (2.11 ± 0.61 AU) and normoglycemic rats (1.43 ± 0.48 AU). No difference was found in adherent leukocytes or in the leukocyte rolling flux between diabetic and normoglycemic rats. We conclude that reactive oxygen species are responsible for an increase in microvascular permeability likely via leukocyte-independent mechanisms.