bluuebunny/crossref_metadata_2025 huggingface dataset:
Influence of Dietary Curcumin, Capsaicin and Garlic on the Antioxidant Status of Red Blood Cells and the Liver in High-Fat-Fed Rats
Authors: Kempaiah R.K. , Srinivasan K.
Published: October 2004

Background/Objective: An animal study was carried out to examine the beneficial influence of known hypolipidemic spice principles – curcumin and capsaicin – and the spice garlic on the antioxidant status of red blood cells and liver tissue in hyperlipidemic rats. Methods: Rats were rendered hyperlipidemic by maintaining them on a high-fat (30%) diet for 8 weeks. Spice principles – curcumin (0.2%) or capsaicin (0.015%) – or garlic (2.0%) were included in the diets of separate animal groups. Erythrocytes isolated at the end of the study were analyzed for intracellular antioxidant molecules and antioxidant enzymes. Results: Intracellular total thiols in the erythrocytes of high-fat-fed rats were depleted significantly (16%). Similarly, the intracellular glutathione content in erythrocytes was depleted in high-fat-fed rats (28%). The concentration of lipid peroxides in the erythrocytes (intracellular as well as membrane) induced by H2O2 was significantly higher in the high-fat-fed group. Curcumin (0.2%) or capsaicin (0.015%) or garlic (2.0%) in the diet which produced the hypotriglyceridemic effect were also effective in reducing oxidant stress, which was indicated by a significant countering of the depleted intracellular antioxidants – total thiols and glutathione – and elevated lipid peroxides in erythrocytes. The elevated lipid peroxide in blood plasma due to the high-fat diet was also significantly countered by the spice treatments. The severely depleted hepatic glutathione in high-fat treatment was also effectively reversed by dietary curcumin, capsaicin and garlic. Conclusions: Thus, dietary hypolipidemic spices were effective in reducing the oxidant stress, which was indicated by countering the depleted antioxidant molecules and antioxidant enzymes in erythrocytes and liver, and decreasing the elevated lipid peroxide content.
The Amount of Dietary Cholesterol Changes the Mode of Effects of (n-3) Polyunsaturated Fatty Acid on Lipoprotein Cholesterol in Hamsters
Authors: Lin Mei-Huei , Lu Shao-Chun , Huang Po-Chao , Liu Young-Chau , Liu Shyun-Yeu
Published: October 2004

This study was designed to investigate the effects of the interaction between dietary (n-3) polyunsaturated fatty acids (PUFA) and different dietary cholesterol content on plasma and liver cholesterol in hamsters. Male Syrian hamsters consumed diets containing an incremental increase in dietary cholesterol content (0, 0.025, 0.05, 0.1 and 0.2%, w/w) with either (n-3) PUFA (21 g/100 g fatty acids) or (n-6) PUFA (37.4 g/100 g fatty acids) fat for 6 weeks. In hamsters fed the nonatherogenic diet (0 or 0.025% dietary cholesterol), very low density lipoprotein (VLDL)-cholesterol levels in the (n-3) PUFA group were not significantly different from those in the (n-6) PUFA group, and low density lipoprotein (LDL)-cholesterol levels in the (n-3) PUFA group were significantly lower than those in the (n-6) PUFA group. In contrast, in hamsters fed the atherogenic diet (0.1 or 0.2% dietary cholesterol), VLDL- and LDL-cholesterol levels in the (n-3) PUFA group were significantly higher than those in the (n-6) PUFA group, in a dose-dependent manner. When the hamsters were fed with 0, 0.025, 0.05, 0.1 or 0.2% (w/w) dietary cholesterol, high density lipoprotein (HDL) cholesterol concentration was significantly lower in the (n-3) PUFA group than those in the (n-6) PUFA group. Hepatic cholesteryl esters were significantly lower, while hepatic microsomal acyl-coenzyme A:cholesterol acyltransferase activity and VLDL-cholesteryl esters were significantly higher in hamsters fed (n-3) PUFA with the atherogenic diet (0.1 or 0.2% dietary cholesterol) than in those fed (n-6) PUFA with the atherogenic diet. Our results demonstrate that the amount of dietary cholesterol is an important factor in determining the mode and extent of effects of dietary (n-3) PUFA, especially on VLDL- and LDL-cholesterol levels. When dietary cholesterol intake was above 0.1% (w/w), the plasma cholesterol-lowering effect of (n-3) PUFA disappeared, and instead, it showed a cholesterol-increasing effect. However, the effects of dietary (n-3) PUFA on HDL-cholesterol are independent of dietary cholesterol content.
Lipoprotein(a) and Other Cardiovascular Metabolic Risk Factors in Kuwaiti Children with Type-1 Diabetes
Authors: Moussa M.A.A. , Alsaeid M. , Abdella N. , Refai T.M.K. , Al-Sheikh N. , Gomez J.E.
Published: October 2004

Background/Aims: Lipoprotein(a) synthesis and catabolism could be influenced by insulin or by diabetes metabolic complications in patients with type-1 diabetes. The aim of the study was to investigate the relation of plasma lipoprotein(a) concentrations with metabolic cardiovascular risk factors in Kuwaiti children with uncomplicated type-1 diabetes. Methods: This case-control study included 115 (44 males and 71 females) diabetic children aged 6–18 years matched by age and sex to 115 non-diabetic children as controls. Results: There was no significant difference between the mean lipoprotein(a) concentrations in type-1 diabetic children (27.34 mg/dl) and their controls (22.80 mg/dl). Total cholesterol, apolipoprotein A1 and B levels were significantly higher in diabetic children than controls. In diabetic children, significant correlations were found between lipoprotein(a) levels and glycated hemoglobin (r = 0.249, p = 0.011), total cholesterol (r = 0.208, p = 0.025), and apolipoprotein B (r = 0.349, p < 0.001). The proportion of diabetic children with lipoprotein(a) >30 mg/dl was significantly higher in those having poor glycemic control (glycated hemoglobin >9.0%, p = 0.013), raised total cholesterol (p = 0.033), or with a family history of cardiovascular disease (p = 0.006). Conclusion: Plasma lipoprotein(a) levels were not elevated in young type-1 diabetic children compared to non-diabetic controls; however, lipoprotein(a) levels were significantly higher in diabetic children with poor glycemic control. Moreover, there were significant correlations between lipoprotein(a) and the metabolic cardiovascular risk factors total cholesterol, atherogenic index, apolipoprotein B and apolipoprotein B/A1 ratio.
Effect of Diabetes mellitus and Different Treatments on Plasma and Erythrocyte Phospholipid Fatty Acid Composition inType 2 Diabetics
Authors: Rodríguez Yanik , Christophe Armand B.
Published: October 2004

Aim: The aim of the present study was to determine how the fatty acid composition of plasma and erythrocyte phospholipids is influenced by the diabetic condition and by dietary or glibenclamide treatment in type 2 diabetes. Methods: The fatty acid composition of plasma and erythrocyte phospholipids is assessed in 3 nonobese groups of Cuban diabetics with comparable age and BMI: 13 newly diagnosed, 15 on diet alone and 15 on diet/glibenclamide. Thirteen healthy individuals were used as controls. Fasting serum glucose, insulin and glycosylated hemoglobin and dietary fat intake were assessed. Results: In plasma phospholipids, significant lower values were found for 14:0, 15:0 and 17:0 in newly diagnosed patients than in controls. In red blood cell phospholipids, 14:1, 20:0 and 24:1 of newly diagnosed patients were significantly lower than of controls, and 16:1ω7 and 18:4ω3 were significantly higher. The different treatments had an effect on 18:0, 18:1t, 20:0 and total trans fatty acids in erythrocyte, but not in plasma phospholipids. Conclusions: Changes in plasma phospholipid fatty acids found in type 2 diabetics may be due to an ‘adapted’ dietary pattern, whereas those in erythrocyte phospholipids may be attributed to effects of the disease itself and may be susceptible to normalization by diet or glibenclamide.
Fibered Confocal Fluorescence Microscopy (Cell-viZio™) Facilitates Extended Imaging in the Field of Microcirculation
Authors: Laemmel Elisabeth , Genet Magalie , Le Goualher Georges , Perchant Aymeric , Le Gargasson Jean-François , Vicaut Eric
Published: October 2004

This study investigated the capability of fibered confocal fluorescence microscopy (FCFM) to provide in vivo microvascular observations. FCFM is specifically designed for in vivo in situ observation thanks to a probe composed of a fiber bundle and micro-optics having a diameter as small as 650 µm. In the first part of the study, we compared the main characteristics of FCFM with those of intravital fluorescence microscopy (IFM). A mouse cremaster preparation was used as a common basis to allow for imaging with both modalities. We discussed the feasibility of obtaining quantitative measurements usually provided by IFM in the context of FCFM: morphometry, capillary permeability, functional capillary density, vasoconstriction and dilation effects. In addition, the possibility to visualize fluorescent red blood cells or leukocytes was also evaluated. Phototoxicity issues and limitations of FCFM were also discussed. We showed that FCFM allows observations and measurements usually provided by IFM and that the real-time capability of the system, as well as the flexibility and small diameter of the optical probe enable micro-invasiveness and can extend imaging capabilities for in vivo in situ observations when compared to IFM.
Effects of Menstrual Cycle on Postoperative Analgesic Requirements, Agitation, Incidence of Nausea and Vomiting after Gynecological Laparoscopy
Authors: Sener Elif Bengi , Kocamanoglu Serhat , Cetinkaya Mehmet Bilge , Ustun Emre , Bildik Emine , Tur Ayla
Published: October 2004

Background: Several studies have suggested that the menstrual cycle has an impact on postoperative nausea and vomiting (PONV). No previous study has evaluated the effect of the menstrual cycle on the incidence of postoperative agitation and analgesic/antiemetic requirements. Methods: On the basis of the phase of the menstrual cycle [pre±menstrual (Pd 25–6), early follicular phase (Pd 8–12), ovulatory phase (Pd 13–15), and luteal phase (Pd 20–24)], 67 patients enrolled in this blinded, prospective study. Anesthesia was standardized. Fentanyl was given to the patients who had severe pain in the recovery room. The patients who had agitation were given midazolam. When pain intensity was >5 on the Visual Analog Scale, metamizol was administered in the Gynecology Department. A blinded anesthesiologist recorded episodes of PONV in the recovery room, and 2 and 24 h postoperatively. Results: The opioid requirement and the frequency of agitation were similar in each group. Metamizol consumption was highest in the luteal phase (p < 0.05). The follicular and luteal phases were predictors for vomiting at recovery (p < 0.05 and p < 0.001, respectively). At the postoperative 2nd hour, nausea was higher in the follicular phase than in the other phases (p < 0.05) and the luteal phase was a predictor for retching (p < 0.001). At the postoperative 24th hour, nausea was the common symptom in the luteal phase (p < 0.05). The need for ondansetron was highest in the luteal phase (p < 0.01). Conclusions: In conclusion, we suggest that the scheduling of all surgical procedures according to the menstrual phase may serve to reduce the incidence of PONV and metamizol/ondansetron consumption and hospital costs.
Nitric Oxide and Effect of a Radical Scavenger N-tert-butyl-α-phenylnitrone on Stroke in a Rat Model
Authors: Saito Kieko , Ikeda Masahiko , Yoshioka Hisashi , Tomita Takako
Published: October 2004

To characterize the role of nitric oxide (NO) in stroke, NO was measured using an in vivo microdialysis technique and electron spin resonance spectrometry in malignant stroke-prone spontaneously hypertensive rats (M-SHRSP), stroke-prone spontaneously hypertensive rats (SHRSP) and normotensive Wistar-Kyoto rats (WKY). The brain dialysate NO level was higher in SHRSP than in WKY. NO was not detected in M-SHRSP hippocampus microdialysate after stroke except after the administration of N-tert-butyl-α-phenylnitrone (PBN). In addition, very little NO was generated in M-SHRSP brain tissue with hemorrhage. These data demonstrate an association between NO and stroke in M-SHRSP. Further, PBN administration results in maintenance of NO levels after stroke in M-SHRSP.
Increased Prevalence of Factor V Leiden Mutation in Premature but Not in Full-Term Infants with Grade I Intracranial Haemorrhage
Authors: Komlósi K. , Havasi V. , Bene J. , Storcz J. , Stankovics J. , Mohay G. , Weisenbach J. , Kosztolányi G. , Melegh B.
Published: October 2004

Objectives: In the current prospective study our aim was to analyse the distribution of the factor V Leiden (G1691A) mutation in preterm and full-term neonates with grade I intraventricular haemorrhage and in control neonates. Study Method: A group of 125 individually selected neonates with grade I intraventricular haemorrhage and 128 controls were investigated. Results: The allele frequency was 7.2% in the total population of affected infants while it was 3.9% in the controls (p < 0.05); the latter corresponds to an average European allele frequency in healthy populations. When the infants were grouped as premature (<2,500 g and ≤36 weeks of gestational age) and appropriate for gestational age full-term infants the statistical analysis revealed an increased prevalence of the mutation in the premature group (10% allele frequency vs. 4.8% in the controls, p < 0.05), and a normal prevalence in the mature group (4.6 vs. 3.1%, respectively); therefore, the overall increase was due to the increase of incidence rate in preterm neonates. Conclusions: These data confirm our previous results and suggest that as the preterm and term infants differ from each other in haemorrhage susceptibility in many clinical particulars, carrying of the mutation has probably also a different impact in premature and in full-term infants with respect to the intraventricular haemorrhage.
Cold Perfluorochemical-Induced Hypothermia Protects Lung Integrity in Normal Rabbits
Authors: Yang S.S. , Jeng M.J. , McShane R. , Chen C.Y. , Wolfson M.R. , Shaffer T.H.
Published: October 2004

To test the hypothesis that intrapulmonary perfluorochemical (PFC) liquid may induce hypothermia, and to compare the effects of internal (IC), external (EC), and combined cooling techniques (EC + IC), 14 juvenile rabbits were randomized to EC by a cold blanket (4°C, n = 5), IC by intrapulmonary cold PFC liquid lavage (4°C, n = 5), or combined IC with PFC and EC (n = 4). Arterial blood gas, blood pressure, and lung mechanics were monitored, and lung histology was examined by light microscopy. The results showed that cooling rates and the time needed to be cooled down to 30°C were significantly faster in EC and EC + IC than IC (p < 0.05). Blood gas analysis and cardiopulmonary function were within the normal range in all groups. Histological assessment revealed varied atelectasis in all lung regions in EC, whereas PFC-filled lungs (IC and EC + IC) demonstrated more homogenous expansion and no evidence of atelectasis. The results indicate that intrapulmonary PFC may be an effective technique to induce and/or augment hypothermia while supporting gas exchange, lung volume and pulmonary architecture.
PDGF and IL-1β Upregulate Cofilin and LIMK2 in Canine Cultured Pulmonary Artery Smooth Muscle Cells
Authors: Bongalon Shaner , Dai Yan-Ping , Singer Cherie A. , Yamboliev Ilia A.
Published: October 2004

Actin cytoskeleton reorganization is regulated by various actin-binding proteins. Cofilin is the principal filament-depolymerizing protein, whose activity is reduced upon phosphorylation by LIMK. Thus, LIMK and cofilin comprise a signal transduction module regulating actin turnover and myogenic tone in healthy vasculature. Novel functions of smooth muscle cells (SMCs) in the hypertensive pulmonary artery, such as increased motility and proliferation, are supported by the actin cytoskeleton. We therefore hypothesized that bioactive peptides that affect these SMC functions may also result in an upregulation of LIMK and cofilin expression. Semiquantitative RT-PCR and immunoblotting indicated that LIMK2 and cofilin mRNA and protein expression is upregulated in canine pulmonary artery SMCs (PASMCs) exposed to PDGF or IL-1β (10 ng/ml). Inhibition of ERK MAPKs (U-0126, 10 µM) or p38 MAPK (PD-169316, 10 µM), but not PI3Ks (LY-294002, 50 µM), reduced LIMK2 and cofilin gene expression stimulated by PDGF or IL-1β. Inhibition of ROCK (Y-27632, 10 µM) reduced only the IL-1β-stimulated LIMK2 and cofilin expression. These novel observations in PASMCs indicate that LIMK2 and cofilin expression can be induced by PDGF or IL-1β. This parallel upregulation of LIMK2 and cofilin may have potentially broad functional significance for the progress of pulmonary artery disease.
Effects of Eplerenone, a Selective Aldosterone Blocker, on the Progression of Left Ventricular Dysfunction and Remodeling in Rats with Dilated Cardiomyopathy
Authors: Wahed Mir Imam Ibne , Watanabe Kenichi , Ma Meilei , Yamaguchi Kenichi , Takahashi Toshihiro , Tachikawa Hitoshi , Kodama Makoto , Aizawa Yushifusa
Published: October 2004

Aldosterone blockade reduces morbidity and mortality in patients with heart failure. We studied the effects of eplerenone, a novel aldosterone blocker, on the progression of left ventricular dysfunction and remodeling in rats with dilated cardiomyopathy after autoimmune myocarditis. Twenty-eight days after immunization, the surviving Lewis rats were randomized to 1 month’s oral treatment with low-dose eplerenone (group L), high-dose eplerenone (group H) or vehicle (group V). Five of 15 (33%) rats in group V and 3 of 15 (20%) rats in group L died during the course of treatment. High-dose eplerenone significantly reduced cardiomyocyte hypertrophy, heart weight and heart weight to body weight ratio. Eplerenone improved left ventricular function in a dose-dependent manner. Central venous pressure and left ventricular end-diastolic pressure were lower, and ±dP/dt and fractional shortening were higher in group H than group V. Eplerenone also attenuated myocardial fibrosis and reduced left ventricular mRNA expressions of TGF-β1 and collagen-III. Our results indicate that treatment with eplerenone improved left ventricular dysfunction and attenuated left ventricular remodeling in rats with heart failure.
Poor Frequency Discrimination Probes Dyslexics with Particularly Impaired Working Memory
Authors: Banai Karen , Ahissar Merav
Published: October 2004

Substantial difficulties in performing simple auditory discriminations were previously found in some individuals with a specific reading disability but not in others. This high variability in psychoacoustic performance raises the question of whether this difficulty is related to the reading deficit. Addressing this question, we compared adult dyslexics with and without difficulty in simple auditory discriminations, using 2-tone frequency discrimination as our probe. The distribution of their frequency discrimination scores was bimodal. On this basis, we divided our participants into subgroups having either poor or adequate psychoacoustic performance. Only dyslexics with poor psychoacoustic scores had significantly impaired verbal working memory compared to their matched controls. Furthermore, and only in this subgroup, working memory scores were correlated with both cognitive abilities and reading-related tasks. Consistent with the hypothesis that in this subgroup poor working memory impedes performance in a broad range of academically related tasks, we found that the majority of dyslexics in this subgroup had more extensive academic difficulties and consequently needed special support in schools. We propose that dyslexics with poor psychoacoustic abilities form a distinct subtype of dyslexia in which the core deficit is not specific to phonological components. For these individuals, poor verbal working memory may be the main impediment to success in academic environments.
Across-Site Threshold Variation in Cochlear Implants: Relation to Speech Recognition
Authors: Pfingst Bryan E. , Xu Li , Thompson Catherine S.
Published: October 2004

Functional implications of across-site variation in detection thresholds in subjects with cochlear implants were evaluated by comparing thresholds to speech recognition performance. Detection thresholds for bipolar (BP) and monopolar (MP) stimulation of all available stimulation sites were assessed in 21 subjects with Nucleus® CI24M and CI24R(CS) implants. We found significant negative correlations between speech recognition and within-subject across-site threshold variance for both BP and MP stimulation, but no significant correlation of speech recognition with mean threshold levels. These results suggest that across-site variance of detection thresholds could provide a useful early indication of the prognosis for speech recognition and might serve as an indicator for specific therapeutic approaches in individual subjects.
Paraquat-Induced Hair Cell Damage and Protection with the Superoxide Dismutase Mimetic M40403
Authors: Nicotera Thomas M. , Ding Dalian , McFadden Sandra L. , Salvemini Daniela , Salvi Richard
Published: October 2004

Some forms of ototoxicity appear to be mediated primarily by the superoxide radical; however, the exact role the superoxide radical plays in cochlear damage is not well understood because most ototoxic drugs produce multiple reactive oxygen species. To characterize the role of the superoxide radical in cochlear damage and the protective effect of compounds that inactivate superoxide, we treated mouse cochlear organotypic cultures for 24 h with paraquat, an herbicide that produces high levels of superoxide. M40403, a highly specific, nonpeptidyl mimetic of superoxide dismutase, was added to some cultures to inactivate the superoxide radical generated by paraquat. The number of outer hair cells (OHC) and inner hair cells (IHC) systematically decreased with increasing concentration of paraquat (0.01–10 mM). M40403 (10 µM) significantly increased OHC and IHC survival in cultures treated with 0.01–1.0 mM of paraquat. These results suggest that excess production of superoxide radical is a sufficient condition for hair cell loss.
Process of Measuring in Clinical Medicine – Implications of Different Definitions in Clinical Therapeutic Studies
Authors: Beckerling Andree , Braun Waldemar , Sommer Michael
Published: October 2004

Often, the decision whether a patient can be considered as healthy or ill is very difficult and depends on measurements and the interpretation of the results. Therefore, it is necessary to build and establish devices that base on certain pre-considerations (who should assess the data, when and how should be measured). Additionally, these devices have to be valid, reliable and responsive. According to the literature we came to the conclusion that no uniform definition of what is ‘healthy’ exists. Referring to mathematical considerations we show an approach to solve this problem especially when the device is a scale. Based on the quality of the scale and of certain pre-considerations normal ranges can be established that will help to distinguish whether patients are ‘healthy‘ or ‘ill’. In this short review we discuss the problem of the establishment of a cut-off level on the basis of devices and we try to point out a solution to solve the arising problems.
Clinical and Radiological Findings in Dominance of Santorini’s Duct
Authors: Kamisawa Terumi , Egawa Naoto , Nakajima Hitoshi , Okamoto Atsutake
Published: October 2004

Background/Aims: Few studies on Santorini’s duct dominance, in which the ventral pancreatic duct is narrower than and anastomoses with Santorini’s duct have been performed. We examined clinical and radiological findings in cases characterized by dominance of Santorini’s duct. Methods: We reviewed 3,800 cases of endoscopic retrograde cholangiopancreatography. Clinical and pancreatographic findings including caliber, course, terminal shape, and patency of Santorini’s duct were examined in cases of Santorini’s duct dominance. Results: Twenty-nine cases were diagnosed as Santorini’s duct dominant. Chronic pancreatitis, acute relapsing pancreatitis, pancreatic-type pain, and hyperamylasemia not associated with obvious pancreatitis were observed in 3, 1, 5, and 6 cases, respectively. Cholangiopancreatographic findings indicated congenital choledochal cyst (n = 2), branch fusion between the ventral and dorsal pancreatic ducts (n = 23), and normal pancreatic duct system characterized by a straight course through the body and tail to join the ventral pancreatic duct in the neck portion of the pancreas (n = 4). Regarding terminal shapes of Santorini’s duct, cudgel type (n = 9) and spindle type (n = 8), which showed frequent patency, were observed significantly more frequently than in controls. Patency of Santorini’s duct was observed in 90% (17/19). Conclusions: Many Santorini’s duct-dominant cases exhibited branch fusion between the ventral and dorsal pancreatic ducts. Although Santorini’s duct functions well in most cases in which it is dominant, pancreatitis or pancreatic-type pain occurs in half of such cases due to relative impairment of function of the minor duodenal papilla.
Pranlukast Inhibits NF-κB Activation and MUC2 Gene Expression in Cultured Human Epithelial Cells
Authors: Ishinaga Hajime , Takeuchi Kazuhiko , Kishioka Chikako , Suzuki Shinya , Basbaum Carol , Majima Yuichi
Published: October 2004

Pranlukast is a selective cysteinyl leukotriene1 (cysLT1) receptor antagonist, and is now widely used in the treatment of asthma. The anti-asthmatic effect of pranlukast may be rendered not only by antileukotriene activity, but also by other pharmacological activity. This study was designed to investigate whether pranlukast had inhibitory effects on nuclear factor-ĸB (NF-ĸB) activation and mucin gene expression in cultured human epithelial cells. Luciferase assay was mainly used for analysis. Cultured epithelial cells were transfected with NF-ĸB luciferase vector, MUC2 or MUC5AC luciferase vectors. Lipopolysaccharide (LPS) significantly increased NF-ĸB activation in NCI-H292 cells, which was inhibited by the pretreatment by pranlukast in a dose-dependent manner. Either LTD4 or pranlukast alone did not increase NF-ĸB activation in NCI-H292 cells. Pranlukast also inhibited NF-ĸB activation induced by phorbol 12-myristate 13-acetate (PMA). Pranlukast also significantly inhibited LPS-induced MUC2 mRNA expression by reverse transcription-polymerase chain reaction (RT-PCR) analysis in NCI-H292 cells. Pranlukast also inhibited LPS-induced MUC2 gene expression in HM3-MUC2 cells. However, pranlukast did not inhibit MUC5AC gene transcription activity induced by lipoteichoic acid (LTA) in NCI-H292 cells. These results suggest that pranlukast may inhibit NF-ĸB activation and MUC2 gene transcription through pathways distinct from cysLT1 receptor antagonism in cultured human epithelial cells.
Substitution of Pichia pastoris-Derived Recombinant Proteins with Mannose Containing O- and N-Linked Glycans Decreases Specificity of Diagnostic Tests
Authors: van Oort Erica , Lerouge Patrice , de Heer Pleuni G. , Séveno Martial , Coquet Laurent , Modderman Piet W. , Faye Loïc , Aalberse Rob C. , van Ree Ronald
Published: October 2004

Background: Recombinant proteins from Pichia pastoris need to be fully evaluated before used as diagnostic tools. Objective: The objective of this study was to investigate whether glycosylation by P. pastoris interferes with the specificity of diagnostic tests. Methods: An autoantigen involved in Wegener’s disease (protease 3) and 2 major inhalant allergens from grass pollen (Dac g 5) and house dust mite (Der p 1) were produced as recombinant molecules in P. pastoris. O-linked glycans on Dac g 5 were characterized by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The immune reactivity of the recombinant proteins was compared to that of their natural counterparts by ELISA and a radio-allergosorbent test (RAST) as well as by ELISA and RAST inhibition. Results: In contrast to the non-glycosylated natural allergen, recombinant Dac g 5 was shown to carry at least 2 small mannose-containing O-glycans. We showed that both these O-glycans and the N-linked glycans on recombinant protease 3 and recombinant Der p 1 were recognized in ELISA by IgG antibodies in sera of healthy individuals. These IgG responses were closely correlated. The natural autoantigen and allergens were not recognized by IgG antibodies from healthy subjects. The carbohydrate nature of the epitopes recognized by IgG on the recombinant proteins was confirmed by inhibition studies with mannose and yeast mannan. IgE recognition of yeast glycans was observed in 2 out of 9 positive sera from patients with allergic bronchopulmonary aspergillosis. Conclusion: Production of recombinant molecules in yeast (or moulds) can introduce IgG-binding glycans that negatively affect the specificity of diagnostic tests.
Mast Cell Tryptase Activates Peripheral Blood Eosinophils to Release Granule-Associated Enzymes
Authors: Vliagoftis Harissios , Lacy Paige , Luy Betty , Adamko Darryl , Hollenberg Morley , Befus Dean , Moqbel Redwan
Published: October 2004

Background: Mast cells and eosinophils are important effector cells in asthma. Understanding their interactions is essential for studying asthma pathophysiology. Inflammatory mediators released from mast cells, such as arachidonic acid metabolites, TNF and IL-5, are important in eosinophil biology. However, little is known about the effects of mast cell-specific mediators, such as tryptase, on eosinophils. Our objective was to investigate the effects of mast cell tryptase on human peripheral blood eosinophils. Methods: Peripheral blood eosinophils isolated from asthmatic individuals were activated using various concentrations of tryptase- and protease-activated receptor-2 (PAR-2)-activating peptides (PAR-2 AP). Eosinophil activation was evaluated by the release of granule mediators, superoxide release, estimation of eosinophil survival, changes in intracellular Ca2+ concentration and mitogen-activated protein kinase activation. Results: Tryptase induced the release of eosinophil peroxidase and β-hexosaminidase from peripheral blood eosinophils but had no effect on RANTES release. Eosinophils isolated from two thirds of our donors responded to tryptase, while the remainder appeared not to respond. Release of granule mediators was dependent on tryptase enzymatic activity. To identify the mechanism of eosinophil activation by tryptase, we studied the expression of PAR-2 by eosinophils and its function. Using RT-PCR, we amplified PAR-2 from eosinophils. However, flow cytometry failed to detect significant PAR-2 expression on the surface of eosinophils. The PAR-2 AP SLIGRL-NH2 did not induce eosinophil activation by any of the methods we employed. Conclusion: Our data indicate that mast cell tryptase may affect eosinophil activation status independently of PAR-2.
The Anti-Allergic Effects of Lactic Acid Bacteria Are Strain Dependent and Mediated by Effects on both Th1/Th2 Cytokine Expression and Balance
Authors: Fujiwara Daisuke , Inoue Sayo , Wakabayashi Hideyuki , Fujii Toshio
Published: October 2004

Background: There is growing interest in the immune-stimulating effect and in particular, the anti-allergic effect, of lactic acid bacteria (LAB). However, no comprehensive studies have been done that compare the immune-stimulating potential of LAB strains. Methods: The in vitro immune-stimulating effects on Th1/Th2 balance of more than 100 LAB strains were compared in splenocytes from ovalbumin-sensitized Th2-polarized mice. The in vivoanti-allergic ability of strain KW3110 was studied in the Th2-polarized model by detecting serum IgE concentration, Th1/Th2 cytokine secretion from splenocytes, and the expression of co-stimulatory molecules on macrophages. Results: In vitro studies from Th2-polarized splenocytes, using IL-12 as a Th1 parameter and IL-4 secretion as a Th2 parameter revealed a wide variety of IL-12-inducing and IL-4-repressing activities, depending on the strain of LAB, not depending on the species. However, evaluation of individual strains in vivo revealed that after exposure to Lactobacillus paracasei KW3110 strain, the serum IgE elevation elicited by repeated OVA injection of mice was strongly inhibited. Cytokine secretion from splenocytes 20 weeks after KW3110 administration showed increased IL-12 and decreased IL-4 expression. Both CD40 and B7-1 expression on macrophages was upregulated by administration of KW3110. Conclusions: Improving the consequences of the Th1/Th2 imbalance by administration of LAB was dependent upon the LAB strain rather than the LAB species. Oral KW3110 administration in the mouse allergy model directed the Th1/Th2 balance toward Th1 through the maturation of APCs and inhibition of serum IgE elevation.
Lipopolysaccharide Contamination of β-Lactoglobulin Affects the Immune Response against Intraperitoneally and Orally Administered Antigen
Authors: Brix Susanne , Kjær Tanja M.R. , Barkholt Vibeke , Frøkiær Hanne
Published: October 2004

Background: Microbial components in the environment are potent activators of the immune system with capacity to shift the active immune response towards priming of Th1 and/or Th2 cells. Lipopolysaccharide (LPS), a cell-wall component of Gram-negative bacteria, is extensively present in food products like cow’s milk. It is not well established, however, how this presence of LPS affects oral tolerance induction. Methods: We studied the effect of LPS contamination in a commercial preparation of the cow milk protein β-lactoglobulin (β-LG) on antigen-specific immune responses. IgG1/IgG2a production upon intraperitoneal immunization without adjuvant was measured, and oral tolerance induction against β-LG after administration of either an aqueous solution or water-in-oil (w/o) emulsion of β-LG was evaluated. Results: LPS contamination of β-LG provoked a β-LG-specific IgG2a response, as well as an enhanced β-LG-specific IgG1 response upon intraperitoneal immunization. Oral tolerance induction to β-LG was induced by aqueous solutions of β-LG with and without LPS administration. Conversely, oral administration of w/o-emulsified β-LG prevented oral tolerance to β-LG only when the β-LG was contaminated with LPS. Conclusions: LPS contamination of an aqueous protein solution does not affect oral tolerance induction, whereas LPS present in emulsion prevents oral tolerance induction towards the food protein.
Relationship between Active and Passive Smoking and Total Serum IgE Levels in Japanese Women: Baseline Data from the Osaka Maternal and Child Health Study
Authors: Miyake Y. , Miyamoto S. , Ohya Y. , Sasaki S. , Matsunaga I. , Yoshida T. , Hirota Y. , Oda H.
Published: October 2004

Background: Many studies have shown that cigarette smoking is associated with elevated concentrations of total serum IgE. Few studies, however, have examined total IgE in relation to passive smoking exposure, especially in adults. This cross-sectional study investigated the association of active and passive smoking exposure with levels of total serum IgE in Japan. Methods: Study subjects were 981 pregnant women in Osaka. Total IgE levels were measured using UniCAP 1000 and were defined as elevated if they exceeded 170 ml/UI. Age, gestation, parity, family history of asthma, atopic eczema and allergic rhinitis, indoor domestic pets, family income, education and the mite allergen level in house dust were selected as potential confounding factors. Results: Current smoking of at least 15 cigarettes a day and 8.0 or more pack-years of smoking were independently related to an increased prevalence of elevated total serum IgE (adjusted odds ratios 3.40 and 2.51, 95% confidence intervals 2.12–5.47 and 1.55–4.06, respectively), and both cigarette smoking status and pack-years of smoking were significantly positively associated with total serum IgE levels, especially in subjects with a positive familial allergic history. There was no measurable association of exposure to environmental tobacco smoke (ETS) at home or at work with total serum IgE concentrations among those who had never smoked. Conclusions: Our results corroborate a positive relationship between active smoking and total serum IgE levels; however, this study failed to substantiate a positive association of ETS exposure with total IgE. Investigations with more precise and detailed exposure measurements are warranted.
Is the Mycobacteria-Derived Purified Protein Response in Atopic Asthmatic Children Different?
Authors: Kale Hüseyin Serdar , Taştan Yücel , Pinçe Osman , Altuncu Emel , Erginöz Ethem
Published: October 2004

Background: The response to mycobacteria-derived purified protein (PPD) is mediated primarily by T-helper-1 response and is expected to be inhibited in atopic diseases. The aim of this study was to investigate whether the PPD response is different in atopic asthmatic children. Methods: 40 atopic asthmatic children (mean age 8.3 ± 4.9 years) and 40 healthy age- and sex-matched children who had received bacillus Calmette-Guérin (BCG) vaccination were included in the study. Five PPD units were administered intradermally to all children and were interpreted after 48 h. Results: There was no correlation between serum total IgE level and PPD induration (p = 0.054). The PPD induration was not statistically different between the children who used inhaled corticosteroid and those who did not. Although the PPD positivity (induration ≧5 mm) rate was higher in atopic asthmatic children (50%) than in healthy children (32.5%), the difference was not found to be statistically significant. The PPD induration in atopic asthmatic children (7.41 ± 5.58 mm) was found to be greater than the one in healthy children (5.21 ± 3.39) (p < 0.039). The induration in atopic asthmatic children (5.21 ± 3.77) and healthy children (4.43 ± 2.32) did not show a difference in children who where vaccinated only once with BCG, but it was found to be statistically significantly greater in atopic asthmatic children (12.50 ± 5.90) than healthy children (7.08 ± 4.70) who were vaccinated with BCG twice (p < 0.012). The proportion of having a PPD induration of ≧10 mm was found to be higher in atopic asthmatic children than in the healthy ones (32.5 vs. 12.5%) (p < 0.032). Conclusion: Our data showed that the PPD response was stronger in BCG-vaccinated atopic asthmatic children than in healthy BCG-vaccinated ones.
Synthesis and Secretion of Interleukin-15 by Freshly Isolated Human Bronchial Epithelial Cells
Authors: Ge Ning , Nishioka Yasuhiko , Nakamura Yoichi , Okano Yoshio , Yoneda Kazuo , Ogawa Hirohisa , Sugita Akemi , Yanagawa Hiroaki , Sone Saburo
Published: October 2004

Background: Interleukin-15 (IL-15), which shares many functional activities of IL-2, is proposed as a potential modulator of T and natural killer (NK) cell-mediated inflammatory diseases. Since IL-15 gene is expressed in various cell types including epithelial cells, we examined how proinflammatory modulators affect IL-15 gene expression in both freshly isolated human bronchial epithelial cells (HBECs) and the human bronchial epithelial cell line BEAS-2B. Methods: HBECs were obtained from 25 patients with primary lung cancer by bronchial brushing under bronchofiberscopy. The expressions of IL-15 and its receptor were examined using reverse transcription-polymerase chain reaction (RT-PCR), Northern blot analysis and enzyme-linked immunosorbent assay. Results: IL-15 mRNA was constitutively expressed in the cells and was upregulated by several proinflammatory cytokines such as IL-1β, tumor necrosis factor-α, interferon-γ (IFN-γ) and lipopolysaccharide. In addition, IFN-γ but not other cytokines induced the synthesis and secretion of IL-15 protein. Investigation of IL-15 receptor expression using RT-PCR showed that IL-15Rα and IL-2Rβ chains but not IL-2Rα or γ chain were constitutively expressed in these cells. Conclusions: Bronchial epithelial cells may contribute to T and NK cell-mediated airway inflammation through IL-15 production.
A Normal Organ Antigen and Its Corresponding Antibodies in Pathological Human Sera
Authors: Milgrom Felix , Czechowski Donna
Published: October 2004

Previous studies on the appearance in pathological human sera of antibodies to an antigen of normal mammalian organs were continued. In gel precipitation reactions, antibodies combining with saline extracts of mammalian organs were found in 20 of 63 cancer sera and in 4 of 15 syphilis sera, but only in 3 of 56 other pathological sera. Furthermore, an identical antigen was demonstrated in 5 of 58 pathological sera. The antigen under study did not belong to any group of known heterophile antigens and it was devoid of organ and tissue specificity.
Tumor Necrosis Factor-Alpha-Induced Nitric Oxide Restrains the Apoptotic Response of Anterior Pituitary Cells
Authors: Candolfi Marianela , Jaita Gabriela , Zaldivar Verónica , Zárate Sandra , Pisera Daniel , Seilicovich Adriana
Published: October 2004

We previously reported that tumor necrosis factor-α (TNF-α) inhibits cell proliferation whereas it stimulates apoptosis of anterior pituitary cells in an estrogen-dependent manner. Also, we showed that nitric oxide (NO) mediates the inhibitory effect of TNF-α on prolactin release. Here, we studied the effect of TNF-α on nitric oxide synthase (NOS) activity and expression in anterior pituitary cells from cycling and ovariectomized (OVX) rats, and the role of NO in TNF-α induced apoptosis of anterior pituitary cells. NOS activity was higher in anterior pituitary cells from rats in proestrus than in diestrus and was stimulated by 17β-estradiol (10–9 M, E2). TNF-α (50 ng/ml) stimulated NOS activity in anterior pituitary cells from rats at both stages of the estrous cycle and in cells from OVX rats cultured either with or without E2. Inducible NOS (iNOS) gene expression was higher in anterior pituitary cells from rats in proestrus than in diestrus and its expression was enhanced by TNF-α. Acute administration of E2 to OVX rats increased endothelial NOS (eNOS) expression in the anterior pituitary gland. Also, E2 increased eNOS mRNA in dispersed anterior pituitary cells from OVX rats, and this effect was blocked by TNF-α. nNOS expression in the anterior pituitary gland was higer at proestrus than at diestrus but eNOS expression was similar at both stages. TNF-α decreased eNOS mRNA in anterior pituitary cells from rats at proestrus or diestrus. In anterior pituitary cells from OVX rats, TNF-α failed to induce apoptosis but was able to induce it when cells were incubated with NAME or NMMA, NOS inhibitors that did not affect cell viability per se. In the presence of E2, NAME induced apoptosis and enhanced the proapoptotic effect of TNF-α. In conclusion, our study shows that TNF-α upregulates iNOS gene expression whereas it downregulates estrogen-induced eNOS expression in anterior pituitary cells. Endogenous NO may restrain rather than mediate the proapoptotic effect of TNF-α in anterior pituitary cells.
Mineralocorticoid Treatment Upregulates the Hypothalamic Vasopressinergic System of Spontaneously Hypertensive Rats
Authors: Pietranera Luciana , Saravia Flavia , Roig Paulina , Lima Analía , de Nicola Alejandro F.
Published: October 2004

Mineralocorticoid effects in the brain include the control of cardiovascular functions, induction of salt appetite, interaction with the vasoactive neuropeptides arginine vasopressin (AVP) and angiotensin II and development or aggravation of hypertension. In this regard, mineralocorticoids may play a pathogenic role in rats with a genetic form of hypertension (spontaneously hypertensive rats, SHR). Our objective was to compare the response of the hypothalamic vasopressinergic system to mineralocorticoid administration in SHR and control Wistar-Kyoto (WKY) rats. Sixteen-week-old male SHR showing a systolic blood pressure of 190 ± 5 mm Hg and normotensive WKY rats (130 ± 5 mm Hg) were treated subcutaneously with oil vehicle or a single 10-mg dose of deoxycorticosterone acetate (DOCA). After 2 h, rats were sacrificed and brains prepared for immunocytochemistry of Fos and vasopressin V1a receptor (V1aR) and for non-isotopic in situ hybridization of AVP mRNA. In the basal state, SHR demonstrated a higher number of AVP mRNA- and V1aR-immunopositive cells in the magnocellular division of the paraventricular hypothalamic nucleus (PVN) than WKY rats. After DOCA injection, SHR responded with a significant increase in both parameters with respect to vehicle-injected SHR. In WKY rats, DOCA was without effect on AVP mRNA although it increased the number of V1aR-positive cells. Changes in the number of Fos-positive nuclei were measured in the PVN, median preoptic nucleus (MnPO) and organum vasculosum of the lamina terminalis (OVLT), a circumventricular region showing anatomical connections with the PVN. In vehicle-injected rats, the PVN of SHR showed a higher number of Fos-positive nuclei than in WKY rats, whereas after DOCA treatment, a significant increment occurred in the OVLT but not in the PVN or MnPO of the SHR group only. These data suggest that the enhanced response of the vasopressinergic system to mineralocorticoids may contribute to the abnormal blood pressure of SHR.
Frameless Localization for Functional Neurosurgical Procedures: A Preliminary Accuracy Study
Authors: Henderson Jaimie M.
Published: October 2004

Background: Targeting for functional stereotactic procedures is traditionally carried out in a noninteractive fashion, without real-time positional feedback. In addition, stereotactic frames are uncomfortable for patients and may impede intraoperative neurological evaluation. As an initial step toward a fully frameless approach to functional surgery, we have investigated the use of an image-guided microdrive coupled to a stereotactic frame system. Methods: For initial laboratory tests, a stereotactic phantom was imaged using high-resolution CT scanning. Three representative targets were chosen within the phantom. Targeting was carried out in the usual fashion using the StealthStation planning suite, utilizing the Radionics CRW system. An LED-equipped reference arc was attached to the CRW base ring. Registration of the base ring was accomplished using a spherical probe. A custom-built microdrive was fitted with an LED array, mounted on the CRW arc and tracked by the StealthStation. The distance between the Stealth real-time localization and the CRW localization was measured on-screen. To evaluate the accuracy of the system in the operating room, a similar procedure was carried out in 13 functional neurosurgical operations (pallidotomy or deep brain stimulator placement). Results: Errors of localization in the laboratory setting ranged from 0.53 to 0.70 mm. In 11 operative cases, the average difference between the CRW localization and the Stealth localization was 1.77 mm. In the remaining 2 cases, equipment malfunction prevented measurement of localization error. Conclusions: Frameless image-guided localization compares favorably to targeting performed noninteractively. In addition, real-time positional feedback confers advantages in target region visualization and confidence in placement of lesions and stimulators for functional procedures. With small improvements in accuracy and system reliability, fully frameless functional procedures could be safely carried out.