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Radioimmunoassay

Distribution by Scientific Domains
Medical Sciences70%
Life Sciences25%
Earth and Environmental Science2%
Chemistry2%
Distribution within Medical Sciences
Dermatology7%
Andrology5%
Diabetes4%
Rheumatology2%
28 Other Subdomains70%

Kinds of Radioimmunoassay

  • solid-phase radioimmunoassay
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    Selected Abstracts

    Dental pulp fibroblasts express neuropeptide Y Y1 receptor but not neuropeptide Y

    INTERNATIONAL ENDODONTIC JOURNAL, Issue 10 2010
    S. A. Killough
    Killough SA, Lundy FT, Irwin CR. Dental pulp fibroblasts express neuropeptide Y Y1 receptor but not neuropeptide Y. International Endodontic Journal, 43, 835,842, 2010. Abstract Aim, To investigate whether dental pulp fibroblasts express neuropeptide Y (NPY) and NPY-Y1 in vitro and to determine the effects of the cytokines including interlukin-1, (IL-1,), TGF- ,1, substance P and NPY on the expression of NPY Y1. Methodology, Three primary fibroblast cell strains were obtained from freshly extracted human third molar teeth. RT-PCR was utilized to detect expression of NPY and mRNA expression. Membrane protein samples were isolated, and protein expression was determined by Western blotting. Radioimmunoassay was used to quantify NPY expression in healthy (n = 35) and carious (n = 39) whole pulp samples, and the student's t -test was used to test for statistical significance. In addition, the 3-(4,5-Dimethylthiazol,2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to assay fibroblast cell growth. Results, mRNA transcripts were found in all three fibroblast cell populations with the cytokines having a stimulatory effect on its expression (P < 0.05). NPY mRNA was not detected in the cell strains. NPY-Y1 receptor protein expression was visualized by Western blotting, and there was no effect of IL-1, or TGF- ,1 on its expression. The mean concentration of NPY-Ir determined by radioimmunoassay in non-carious teeth was 19.40 ng g,1 (±17.03 SD) compared to 29.95 ng g,1 (±20.99 SD) in carious teeth (P < 0.05). Conclusion, Human dental pulp fibroblasts express, but do not synthesize, NPY, demonstrating that the fibroblast is a target cell for NPY. The effect of proinflammatory cytokines suggests that fibroblasts play a neuroimmunomodulatory role in the pulpal response to dental caries and injury. [source]

    Molecular response of nasal mucosa to therapeutic exposure to broad-band ultraviolet radiation

    JOURNAL OF CELLULAR AND MOLECULAR MEDICINE, Issue 1-2 2010
    David Mitchell
    Abstract Ultraviolet radiation (UVR) phototherapy is a promising new treatment for inflammatory airway diseases. However, the potential carcinogenic risks associated with this treatment are not well understood. UV-specific DNA photoproducts were used as biomarkers to address this issue. Radioimmunoassay was used to quantify cyclobutane pyrimidine dimers (CPDs) and (6,4) photoproducts in DNA purified from two milieus: nasal mucosa samples from subjects exposed to intranasal phototherapy and human airway (EpiAirwayÔ) and human skin (EpiDermÔ) tissue models. Immunohistochemistry was used to detect CPD formation and persistence in human nasal biopsies and human tissue models. In subjects exposed to broadband ultraviolet radiation, DNA damage frequencies were determined prior to as well as immediately after treatment and at increasing times post-treatment. We observed significant levels of DNA damage immediately after treatment and efficient removal of the damage within a few days. No residual damage was observed in human subjects exposed to multiple UVB treatments several weeks after the last treatment. To better understand the molecular response of the nasal epithelium to DNA damage, parallel experiments were conducted in EpiAirway and EpiDerm model systems. Repair rates in these two tissues were very similar and comparable to that observed in human skin. The data suggest that the UV-induced DNA damage response of respiratory epithelia is very similar to that of the human epidermis and that nasal mucosa is able to efficiently repair UVB induced DNA damage. [source]

    Assessment of urinary total testosterone production by a highly sensitive time-resolved fluorescence immunoassay ,

    JOURNAL OF CLINICAL LABORATORY ANALYSIS, Issue 6 2008
    Shihua Bao
    Abstract Radioimmunoassay (RIA) that involves purification of the analyte by organic solvent extraction is widely used. Although the extraction RIA is reliable when properly validated, it is time consuming and radioactive, we measure urinary total testosterone with a highly sensitive rapid and accurate time-resolved fluorescence immunoassay (TRFIA) method. High affinity antitestosterone antibody and Eu3+ labeled Donkey antisheep IgG as tracers were used. The assay was evaluated for specificity, sensitivity, analytical recovery, precision and dilution linearity by the TRFIA method on urine samples. A satisfactory standard curve for testosterone TRFIA has been developed with good sensitivity (5.1,pmol/L). The validity of the assay for urinarytotal testosterone was confirmed by thegood correlation between the results obtained by TRFIA (X) and those RIA (Y) (Y=0.075+0.971X, R=0.992). Specificity, analytical recovery, precision and dilution linearity studies were determined and all found to be satisfactory. Male urinary total testosterone excretion ranged from 64.00 to 374.11,nmol/24,hr, which was about four times more than the range for women urinary testosterone excretion (14.16,100.65,nmol/24,hr), which suggests that a direct, reliable, easy to automate, highly sensitive and specific TRFIA type assay for the measurement of total testosterone in urine samples has been developed. J. Clin. Lab. Anal. 22:403,408, 2008. © 2008 Wiley-Liss, Inc. [source]

    Cytokine levels in crevicular fluid are less responsive to orthodontic force in adults than in juveniles

    JOURNAL OF CLINICAL PERIODONTOLOGY, Issue 8 2002
    Yijin Ren
    Abstract Objectives:, Bone remodelling during orthodontic tooth movement is related to the expression of mediators in gingival crevicular fluid (GCF). No information is available concerning the effect of age on the levels of these mediators in GCF. The purpose of this study was to quantify three mediators (prostaglandin E2, interleukin-6 and granulocyte-Macrophage Colony-Stimulating Factor) in GCF during orthodontic tooth movement in juveniles and adults. Material and methods:, A total of 43 juvenile patients (mean age 11 ± 0.7 year), and 41 adult patients (mean age 24 ± 1.6 year) took part in the study. One of the lateral incisors of each patient was tipped labially, the other served as control. GCF samples were taken before force activation (t0) and 24 h later (t24). Mediator levels were determined by radioimmunoassay (RIA). Results: PGE2 concentrations were significantly elevated at t24 in juveniles and adults, while concentrations of IL-6 and GM-CSF were significantly elevated only in juveniles. Total amounts of all three mediators in GCF significantly increased at t24 in both groups. Conclusions:, In early tooth movement, mediator levels in juveniles are more responsive than levels in adults, which agrees with the finding that the initial tooth movement in juveniles is faster than in adults and starts without delay. Zusammenfassung Cytokinlevel in der krevikulären Flüssigkeit sind weniger veränderbar bei Erwachsenen als bei Jugendlichen infolge orthodontischer Kräfte Ziele: Knochenremodellation während orthodontischer Zahnbewegung steht in Beziehung zur Expression von Mediatoren in der gingivalen Sulkusflüssigkeit (GCF). Es gibt keine Informationen bezüglich des Effektes von Alter auf die Level dieser Mediatoren in der GCF. Der Zweck dieser Studie war die Quantifizierung von drei Mediatoren (Prostaglandin E2, Interleukin-6 und Granulozyten-Makrophagen-Kolonien-stimulierender Faktor) in der GCF während orthodontischer Zahnbewegung bei Jugendlichen und Erwachsenen. Material und Methoden: Insgesamt 43 jugendliche Patienten (mittleres Alter 11±0.7 Jahre) und 41 erwachsene Patienten (mittleres Alter 24±1.6 Jahre) nahmen an der Studie teil. Einer der lateralen Schneidezähne jedes Patienten wurde nach labial gekippt, der andere diente als Kontrolle. GCF Proben wurden vor der Aktivation (t0) und 24 h später (t24) gewonnen. Die Mediatorenlevel wurden mit Radioimmunoassay (RIA) bestimmt. Ergebnisse: Die PGE2 Konzentrationen waren signifikant zum Zeitpunkt t24 bei Jugendlichen und Erwachsenen erhöht, während die Konzentrationen von IL-6 und GM-CSF nur bei Jugendlichen signifikant erhöht waren. Die totale Menge der drei Mediatoren in der GCF nahmen signifikant zum Zeitpunkt t24 in beiden Gruppen zu. Schlussfolgerungen: In der frühen Zahnbewegung sind die Mediatorenlevel bei Jugendlichen mehr veränderbar als bei Erwachsenen. Dies stimmt überein mit der Beobachtung, dass die initiale Zahnbewegung bei Jugendlichen schneller als bei Erwachsenen ist und ohne Verspätung beginnt. Résumé Les niveaux de cytokine dans le fluide gingival répondent moins aux forces orthodontiques chez l'adulte que chez les jeunes. Objectifs: Le remodelage osseux lors des mouvements dentaires orthodontiques est en relation avec l'expression des médiateurs dans le fluide gingival (GCF). Aucune information n'est disponible sur les effets de l'âge sur les niveaux de ces médiateurs dans le GCF. Le but de cette étude était de quantifier 3 médiateurs (la prostaglandine E-2, L'interleukine-6 et le facteur de stimulation des colonies de macrophage et de granulocyte) dans le GCF pendant un mouvement orthodontique chez des jeunes et chez des adultes. Matériels et méthodes: 43 jeunes patients (en moyenne âgés de 11±0.7 ans) et 41 patients adultes (en moyenne âgés de 24±1.6 ans) prirent part à cette étude. Une des incisives latérale de chaque patient fut versée vestibulairement, l'autre servant de contrôle. Des échantillons de GCF furent prélevés avant l'activation de la force (t0) et 24 heures plus tard (t24). Le niveau de cytokine fut déterminé par radioimmunoassay. Résultats: Les concentrations de PgE-2 étaient significativement élevées à t24 chez les jeunes et les adultes, alors que les concentrations de IL-6 et de GM-CSF étaient significativement augmentées seulement chez les jeunes. La quantité totale des 3 médiateurs dans le GCF augmentait significativement à t24 dans chaque groupe. Conclusions: Rapidement après l'application des mouvements dentaires, les niveaux de médiateurs chez les jeunes répondent plus que chez l'adulte, ce qui est en accord avec les résultats qui montrent que le mouvement initial dentaire chez les jeunes est plus rapide que chez les adultes et débute sans délai. [source]

    Granulocyte elastase, matrix metalloproteinase-8 and prostaglandin E2 in gingival crevicular fluid in matched clinical sites in smokers and non-smokers with persistent periodontitis

    JOURNAL OF CLINICAL PERIODONTOLOGY, Issue 5 2002
    B. Söder
    Abstract Background/aims: Smokers with persistent periodontitis may have granulocytes with impaired function. This study aimed to determine the levels of granulocyte elastase, matrix metalloproteinase-8 (MMP-8) and prostaglandin E2 (PGE2) in gingival crevicular fluid (GCF) in smokers and non-smokers with persistent periodontitis. Methods: We analyzed GCF from 70 matched sites in 29 periodontitis and 6 gingivitis sites in 34 subjects, 17 smokers, and 17 non-smokers. We also analyzed separately GCF from 28 of these subjects, 14 smokers and 14 non-smokers in 14 matched periodontitis sites. The following measurements were made: elastase complexed to ,1 -antitrypsin (EA-,1AT) and MMP-8 with ELISA, functional elastase with a chromogenic substrate, and PGE2 with radioimmunoassay (125I RIA). The significance of the findings was determined with Mann-Whitney test. Results: In the 29 matched periodontitis sites, smokers had significantly more functional elastase (p<0.005) and more EA-,1AT (p<0.05) than non-smokers. In the 14 matched periodontitis sites in 14 smokers and 14 non-smokers, the former had significantly more functional elastase than the latter (p<0.001). A significant correlation was found between EA-,1AT and MMP-8 in smokers (p<0.05) and non-smokers (p<0.001) and a positive correlation between levels of functional elastase and MMP-8 in non-smokers (r2=0.98; p<0.001). Conclusions: Granulocyte function seems to be impaired in smokers with persistent periodontitis. The cells react to the bacterial challenge by releasing serine proteases, which reflect the degradation of connective tissue. The risk of progression of the disease is therefore higher in smokers with persistent periodontitis than in non-smokers. Zusammenfassung Hintergrund, Ziele: Raucher mit bestehender Parodontitis haben möglicherweise Granulozyten mit beeinträchtigter Funktion. Diese Studie zielt auf die Bestimmung der Levels von Granulozytenelastase, Matrix-Metalloproteinase-8 (MMP-8) und Prostaglandin E2 (PGE2) in der krevikulären gingivalen Flüssigkeit (GCF) bei Rauchern und Nichtrauchern mit bestehender Parodontitis. Methoden: Wir analysierten GCF von 70 entsprechenden Flächen bei 29 Parodontitis und 6 Gingivitisflächen von 34 Personen, 17 Rauchern und 17 Nichtrauchern. Wir analysierten zusätzlich getrennt die GCF von 28 dieser Personen: 14 Raucher und 14 Nichtraucher von 14 entsprechenden parodontalen Flächen. Die folgenden Messungen wurden vorgenommen: Elastasekomplex zu ,1 -Antitrypsin (EA-,1AT) und MMP-8 mit ELISA, funktionelle Elastase mit chromogenem Substrat und PGE2 mit Radioimmunoassay (125I RIA). Die Signifikanz der Ergebnisse wurde mit dem Mann-Whitney Test bestimmt. Ergebnisse: In den 29 entsprechenden parodontalen Flächen hatten die Raucher signifikant mehr funktionelle Elastase (p<0.005) und mehr EA-,1At (p<0.05) als Nichtraucher. Bei den 14 entsprechenden parodontalen Flächen der 14 Raucher und 14 Nichtraucher hatten die ersten signifikant mehr funktionelle Elastase als die letzteren (p<0.001). Eine signifikante Korrelation wurde zwischen EA-,1AT und MMP-8 bei Rauchern (p<0.05) und Nichtrauchern (p<0.001) gefunden und eine positive Korrelation zwischen den Levels der funktionellen Elastase und MMP-8 bei Nichtrauchern (r2=0.98; p<0.001) festgestellt. Schlussfolgerungen: Die Granulozytenfunktion scheint bei Rauchern mit bestehender Parodontitis beeinträchtigt zu sein. Die Zellen reagieren auf die bakterielle Herausforderung durch Freisetzung von Serinproteasen, die die Degradation von Bindegewebe reflektiert. Das Risiko einer Progression dieser Erkrankung ist deshalb bei Rauchern mit bestehender Parodontitis höher als bei Nichtrauchern. Résumé Origine, but: Les fumeurs avec parodontite persistante pourraient avoir des granulocytes ayant des fonctions déréglées. Cette étude a eu pour but de déterminer les niveaux d'élastase granulocytaire, de la métallo-protéinase-8 de la matrice (MMP-8) et de la prostaglandine E2 (PGE2) dans le fluide créviculaire gingival (GCF) chez les fumeurs et les non-fumeurs avec parodontite persistante. Méthodes: Le GCF a été prélevé de 70 sites équivalents dans 29 parodontites et 6 sites avec gingivite chez 34 sujets, 17 fumeurs et 17 non-fumeurs. Le GCF de 28 de ces sujets a été analysé séparément, 14 fumeurs et 14 non-fumeurs dans 14 sites équivalents du point de vue parodontite. Les mesures suivantes ont été relevées: l'élastase avec ,1 -antitrypsine (EA-,1AT) et MMP-8 par ELISA, l'élastase fonctionnelle avec un substrat chromogénique, et PGE2 avec un essai radio-immunitaire (125I RIA). La signification de ces découvertes a été par l'utilisation du test de Mann-Whitney. Résultats: Dans les 29 sites équivalents, les fumeurs avaient significativement plus d'élastase functionnelle (p<0.005) et plus de EA-,1AT (p<0.05) que les non-fumeurs. Dans les 14 sites équivalents du point de vue parodontite, les 14 fumeurs avaient significativement plus d'élastase fonctionnelle que les 14 non-fumeurs (p<0.001). Une relation significative a étéétablie entre EA-,1AT et MMP-8 chez les fumeurs (p<0.05) et les non-fumeurs (p<0.001) et une relation positive entre les niveaux d'élastase fonctionnelle et de MMP-8 chez les non-fumeurs (r2=0.98; p<0.001). Conclusions: La fonction granulocytaire semble être altérée chez les fumeurs avec parodontite persistante. Les cellules réagissent à l'attaque bactérienne en relâchant des protéases sérine, ce qui démontre une dégradation du tissu conjonctif. Le risque de progression de la maladie est ainsi plus élevé chez les fumeurs avec parodontite persistante que chez les non-fumeurs. [source]

    Influence of Serotonin on the Development and Migration of Gonadotropin-Releasing Hormone Neurones in Rat Foetuses

    JOURNAL OF NEUROENDOCRINOLOGY, Issue 6 2003
    T. Pronina
    Abstract This study used a pharmacological approach to evaluate the consequences of the metabolic perturbations of neurotransmitters on brain development. Pregnant rats received p-chlorophenylalanine (pCPA), an inhibitor of serotonin (5-hydroxytryptamine, 5-HT) synthesis, or saline (control) from the 11th day of gestation once or daily up to the 15th, 17th and 20th day, followed by processing of the forebrain and/or nasal cranium of foetal males and females for high-performance liquid chromatography of monoamines, radioimmunoassay of gonadotropin-releasing hormone (GnRH) and quantitative and semiquantitative immunocytochemistry for GnRH. The pCPA treatment resulted in a 50,70% depletion of 5-HT in the nasal crania and forebrains at any studied age. Radioimmunoassay showed no change in GnRH content in 5-HT deficient foetuses at E16 compared to controls, being higher in both cases in the rostral forebrain than in the hypothalamus. In controls at E21, the GnRH content in the hypothalamus exceeded that in the rostral forebrain, whereas in the 5-HT deficient group the opposite was found. These data suggest that 5-HT provided a stimulating effect on GnRH neurone migration, and this was confirmed by quantification of GnRH-immunoreactive neurones in the forebrain along the trajectory of their migration. At E18 and E21, the fractions of GnRH neurones in the rostral part of the trajectory in pCPA-treated foetuses were greater than those in control foetuses but the opposite was true for the caudal part of the trajectory. Moreover, 5-HT appeared to control the proliferation of the precursor cells of GnRH neurones and their differentiation, as derived from the observations of the increased number of GnRH neurones in the forebrain of foetuses of both sexes, as well as the region-specific decreased neuronal size and content of GnRH in 5-HT-deficient females. Thus, 5-HT appears to contribute to the regulation of the origin, differentiation and migration of GnRH neurones. [source]

    Vasoconstrictor activity of novel endothelin peptide, ET-1(1 , 31), in human mammary and coronary arteries in vitro

    BRITISH JOURNAL OF PHARMACOLOGY, Issue 6 2001
    Janet J Maguire
    The ability of the putative chymase product of big endothelin-1 (big ET-1), ET-1(1 , 31), to constrict isolated endothelium-denuded preparations of human coronary and internal mammary artery was determined. pD2 values in coronary and mammary artery respectively were 8.21±0.12 (n=14) and 8.55±0.11 (n=12) for ET-1, 6.74±0.11 (n=16) and 7.10±0.08 (n=16) for ET-1(1 , 31) and 6.92±0.10 (n=15) and 7.23±0.11 (n=12) for big ET-1. ET-1(1 , 31) was significantly less potent than ET-1 (P<0.001, Student's t -test) and equipotent with big ET-1. Vasoconstrictor responses to 100 , 700 nM ET-1(1 , 31) were significantly (P<0.05, Student's paired t -test) attenuated by the ETA antagonist PD156707 (100 nM). There was no effect of the ECE inhibitor PD159790 (30 ,M), the ECE/NEP inhibitor phosphoramidon (100 ,M) or the serine protease inhibitor chymostatin (100 ,M) on ET-1(1 , 31) responses in either artery. Radioimmunoassay detected significant levels of mature ET in the bathing medium of coronary (1.6±0.5 nM, n=14) and mammary (2.1±0.6 nM, n=14) arteries, suggesting that conversion of ET-1(1 , 31) to ET-1 contributed to the observed vasoconstriction. ET-1(1 , 31) competed for specific [125I]-ET-1 binding to ETA and ETB receptors in human left ventricle with a pooled KD of 71.6±7.0 nM (n=3). Therefore, in human arteries the novel peptide ET-1(1 , 31) mediated vasoconstriction via activation of the ETA receptor. The conversion of ET-1(1 , 31) to ET-1, by an as yet unidentified protease, must contribute wholly or partly to the observed constrictor response. Chymase generated ET-1(1 , 31) may therefore represent an alternative precursor for ET-1 production in the human vasculature. British Journal of Pharmacology (2001) 134, 1360,1366; doi:10.1038/sj.bjp.0704384 [source]

    2127: Ghrelin concentration in the aqueous humour and plasma in open angle glaucoma patients

    ACTA OPHTHALMOLOGICA, Issue 2010
    A KATSANOS
    Purpose Ghrelin is a peptide hormone that exerts metabolic and smooth muscle-relaxant effects in ocular tissues. The aim of this study was to compare aqueous humor and plasma levels of ghrelin in patients with open angle glaucoma (OAG) and controls. Methods Twenty four OAG, including 7 pseudoexfoliation (PXG) and 17 primary open-angle glaucoma (POAG) patients, and 30 controls were included. All participants were patients scheduled for cataract or glaucoma surgery. Patients with other concomitant ocular disease, previous ocular surgery or diabetes were excluded. Blood samples were collected before cataract surgery. Aqueous humor was aspirated from the anterior chamber through a paracentesis with a 27 G needle under sterile conditions. Ghrelin levels in both samples were measured quantitatively with commercially available Radioimmunoassay (RIA) kits. Results Mean±SD age was 71.0±9.3 and 69.6±6.6 years in the OAG and control groups, respectively (p=0.6). Plasma levels of ghrelin were 495.6±157.7 pg/ml in the OAG and 482.2±125.4 pg/ml in the control group, respectively (Mann-Whitney test, p=0.9). Aqueous humor levels of ghrelin were 85.5±15.4 pg/ml and 123.4 ±25.5 pg/ml in the OAG and control groups, respectively (Mann-Whitney test, p<0.01). The ratio of plasma/aqueous concentration in ghrelin was higher in the OAG versus the control group (5.82± 1.94 versus 4.00±1.04, Mann-Whitney test, p<0.01). There was no difference neither in plasma nor in aqueous humor levels of ghrelin between POAG and PXG patients (p>0.5). Conclusion Aqueous humor levels of ghrelin were significantly lower in OAG patients. This difference may manifest a role of ghrelin in the disease process or a consequence of antiglaucoma treatment. [source]

    Prostaglandin I2 sensory input into the enteric nervous system during distension-induced colonic chloride secretion in rat colon

    ACTA PHYSIOLOGICA, Issue 3 2010
    J. D. Schulzke
    Abstract Aim:, Intestinal pressure differences or experimental distension induce ion secretion via the enteric nervous system, the sensorial origin of which is only poorly understood. This study aimed to investigate sensorial inputs and the role of afferent and interneurones in mechanically activated submucosal secretory reflex circuits. Methods:, Distension-induced rheogenic chloride secretion was measured as increase in short-circuit current 10 min after distension (,ISC10; distension parameters ± 100 ,L, 2 Hz, 20 s) in partially stripped rat distal colon in the Ussing-chamber in vitro. PGE2 and PGI2 were measured by radioimmunoassay. Results:, ,ISC10 was 2.0 ± 0.2 ,mol h,1 cm,2 and could be attenuated by lobeline, mecamylamine and dimethylphenylpiperazine, indicating an influence of nicotinergic interneurones. Additionally, a contribution of afferent neurones was indicated from the short-term potentiation of ,ISC10 by capsaicin (1 ,m). As evidence for its initial event, indomethacin (1 ,m) inhibited distension-induced secretion and the release of PGI2 was directly detected after distension. Furthermore, serotoninergic mediation was confirmed by granisetron (100 ,m) which was functionally localized distally to PGI2 in this reflex circuit, as granisetron inhibited an iloprost-induced ISC, while indomethacin did not affect serotonin-activated ion secretion. Conclusions:, Distension-induced active electrogenic chloride secretion in rat colon is mediated by a neuronal reflex circuit which includes afferent neurones and nicotinergic interneurones. It is initiated by distension-induced PGI2 release from subepithelial cells triggering this reflex via serotoninergic 5-HT3 receptor transmission. Functionally, this mechanism may help to protect against intestinal stasis but could also contribute to luminal fluid loss, e.g. during intestinal obstruction. [source]

    Imidazoline-induced amplification of glucose- and carbachol-stimulated insulin release includes a marked suppression of islet nitric oxide generation in the mouse

    ACTA PHYSIOLOGICA, Issue 3 2009
    S. Meidute-Abaraviciene
    Abstract Aim:, The role of islet nitric oxide (NO) production in insulin-releasing mechanisms is unclear. We examined whether the beneficial effects of the imidazoline derivative RX 871024 (RX) on ,-cell function might be related to perturbations of islet NO production. Methods:, Experiments were performed with isolated islets or intact mice challenged with glucose or carbachol with or without RX treatment. Insulin was determined with radioimmunoassay, NO generation with high-performance liquid chromatography and expression of inducible NO synthase (iNOS) with confocal microscopy. Results:, RX treatment, in doses lacking effects on basal insulin, greatly amplified insulin release stimulated by the NO-generating secretagogues glucose and carbachol both in vitro and in vivo. RX also improved the glucose tolerance curve. Islets incubated at high glucose levels (20 mmol L,1) displayed increased NO production derived from both neuronal constitutive NO synthase (ncNOS) and iNOS. RX abrogated this glucose-induced NO production concomitant with amplification of insulin release. Confocal microscopy revealed abundant iNOS expression in , cells after incubation of islets at high but not low glucose levels. This was abolished after RX treatment. Similarly, islets cultured for 24 h at high glucose levels showed intense iNOS expression in , cells. This was abrogated with RX and followed by an amplified glucose-induced insulin release. Conclusion:, RX effectively counteracts the negative impact of ,-cell NO generation on insulin release stimulated by glucose and carbachol suggesting imidazoline compounds by virtue of NOS inhibitory properties being of potential therapeutic value for treatment of ,-cell dysfunction in hyperglycaemia and type 2 diabetes. [source]

    Plasma renin in mice with one or two renin genes

    ACTA PHYSIOLOGICA, Issue 4 2004
    P. B. Hansen
    Abstract Aim:, In the present study we have investigated whether the presence of a second renin gene exerts an overriding influence on plasma renin such that mice with two renin genes have consistently higher renin levels than mice with only one renin gene. Methods:, Plasma renin was determined as the rate of angiotensin I generation using a radioimmunoassay (RIA) kit with (plasma renin concentration, PRC) or without (plasma renin activity, PRA) the addition of purified rat angiotensinogen as substrate. Results:, In male 129SvJ, DBA/2 and Swiss Webster mice, strains possessing both Ren-1 and Ren-2, PRC (ng Ang I mL,1 h,1) averaged 178 ± 36, 563 ± 57 and 550 ± 43 while PRA was 2.9 ± 0.5, 3.6 ± 0.8 and 7.8 ± 1.2. In male C57BL/6, C3H and BALB/c mice that express only Ren-1, PRC averaged 426 ± 133, 917 ± 105 and 315 ± 72, and PRA was 3.4 ± 1.0, 6.9 ± 1.7 and 4.5 ± 1.2. In the two renin gene A1AR,/, mice compared with the one renin gene A1AR+/+, PRC averaged 538 ± 321 and 415 ± 159 while PRA averaged 3.2 ± 1.1 and 4.4 ± 1.4 ng Ang I mL,1 h,1. Aldosterone levels showed no significant differences between one renin (C57BL/6, C3H and BALB/c) and two renin (129SvJ, DBA/2 and Swiss Webster) gene mice. Furthermore, by quantitative real-time polymerase chain reaction (RT-PCR) we found no correlation between the number of renin genes and whole kidney renin mRNA levels from one and two renin gene mice. Conclusion:, Our data show that baseline plasma renin is not systematically higher in mice with two renin genes than in one renin gene mice. Thus, the presence of a second renin gene does not seem to be a major determinant of differences in PRC between different mouse strains. [source]

    Flow cytometry versus histamine release analysis of in vitro basophil degranulation in allergy to Hymenoptera venom

    CYTOMETRY, Issue 1 2003
    C. Lambert
    Abstract Background Flow cytometry (FCM) has been proposed for specific allergy in vitro testing. We investigated its biological significance for allergy to Hymenoptera venoms and compared it with the routinely performed basophil histamine release test (HRT). Methods Blood samples from 26 allergic and 8 nonallergic donors were incubated with venom at serial concentrations. Basophils were analyzed with anti-CD45-PE-Cyanin 5, Anti-IgE-FITC, and Anti-CD63-Phycoerythrine. HRT was measured by radioimmunoassay. Results FCM was as convenient as HRT for measuring basophil reactivity in at least 87% of allergic and 75% of nonallergic subjects. CD63 outer expression was specifically induced in 91% of releaser subjects (86% on HRT) and in 1 of 10 tests in nonallergic donors, or one of six tests (16% on HRT) in allergic patients tested with an irrelevant allergen. Both methods were concordant in 85.7% of the tests. The three discordant patients had low-grade reactions and borderline biological responses on FCM (n = 2) or HRT (n = 1). Conclusions The dynamic, physiologic significance of CD63, the dose,response curve, and dependency on ethylene-diaminetetra acetic acid suggested that both tests reflect the same mechanism. Cytometry Part B (Clin. Cytometry) 52B:13,19, 2003. © 2003 Wiley-Liss, Inc. [source]

    Cardiac natriuretic peptides and continuously monitored atrial pressures during chronic rapid pacing in pigs

    ACTA PHYSIOLOGICA, Issue 2 2000

    Changes in atrial natriuretic peptide (ANP), N-terminal proatrial natriuretic peptide and brain natriuretic peptide (BNP) were evaluated in relation to continuously monitored atrial pressures in a pacing model of heart failure. Pigs were subjected to rapid atrial pacing (225 beats min,1) for 3 weeks with adjustments of pacing frequencies if the pigs showed overt signs of cardiac decompensation. Atrial pressures were monitored by a telemetry system with the animals unsedated and freely moving. Left atrial pressure responded stronger and more rapidly to the initiation of pacing and to alterations in the rate of pacing than right atrial pressure. Plasma natriuretic peptide levels were measured by radioimmunoassay and all increased during pacing with BNP exhibiting the largest relative increase (2.9-fold increase relative to sham pigs). Multiple regression analysis with dummy variables was used to evaluate the relative changes in natriuretic peptides and atrial pressures and the strongest correlation was found between BNP and left atrial pressure with R,2=0.81. Termination of pacing resulted in rapid normalization of ANP values in spite of persistent elevations in atrial pressures. This may reflect an increased metabolism or an attenuated secretory response of ANP to atrial stretch with established heart failure. In conclusion, 3 weeks of rapid pacing induced significant increases in atrial pressures and natriuretic peptide levels. All the natriuretic peptides correlated with atrial pressures with BNP appearing as a more sensitive marker of cardiac filling pressures than ANP and N-terminal proatrial natriuretic peptide. [source]

    Free fatty acids as mediators of adaptive compensatory responses to insulin resistance in dexamethasone-treated rats

    DIABETES/METABOLISM: RESEARCH AND REVIEWS, Issue 2 2008
    Michela Novelli
    Abstract Background Chronic low-dose dexamethasone (DEX) treatment in rats is associated to insulin resistance with compensatory hyperinsulinaemia and reduction in food intake. We tested the hypothesis that the elevation in circulating free fatty acids (FFAs) induced by DEX is the common mediator of both insulin resistance and insulin hyperproduction. Methods For this purpose, an anti-lipolytic agent was administered during DEX treatment to lower lipacidaemia for several hours prior to glucose and insulin tolerance tests. Leptin expression in adipose tissue (by Northern blot) and plasma leptin levels (by radioimmunoassay) were also investigated to verify whether a rise in circulating leptin could be responsible for the anorectic effect of DEX. Results Our data show that a transient pharmacological reduction of elevated plasma FFA levels abates the post-loading hyperinsulinaemia and counteracts the insulin resistance induced by DEX, supporting the hypothesis that the chronic elevation in FFAs is the common mediator of DEX-induced changes. Despite enhanced leptin expression in white adipose tissue, DEX-treated rats show no significant increase in plasma leptin levels. This suggests that the anorectic effect of DEX should be mediated, at least partially, by other factors, possibly related to the influence of concomitantly elevated plasma FFA and insulin levels on the hypothalamic centers regulating feeding. Conclusions Our results sustain the idea that a prolonged increase in plasma FFA levels plays an important role in the adaptive regulation of glucose and energy homeostasis, not only by potentiating insulin secretion but also by providing a signal of ,nutrient abundance' capable of restraining food intake. Copyright © 2007 John Wiley & Sons, Ltd. [source]

    Lack of association between serum adiponectin levels and the Pro12Ala polymorphism in Asian Indians

    DIABETIC MEDICINE, Issue 4 2007
    V. Radha
    Abstract Aims, The aim of the study was to investigate the association of serum adiponectin levels with the Pro12Ala polymorphism of the peroxisome proliferator activated receptor-, (PPARG) gene in Asian Indians. Methods, We selected 400 diabetic subjects, 200 with the Pro12Pro genotype (100 male and 100 female) and 200 with the Pro12Ala genotype (100 male and 100 female) and 400 age- and sex-matched normal glucose tolerance subjects with similar genotype profiles from the Chennai Urban Rural Epidemiology Study. Fasting serum adiponection levels were measured using radioimmunoassay. The Pro12Ala polymorphism was genotyped by PCR,restriction fragment length polymorphism using BstUI. Results, All clinical and biochemical parameters were similar in the subjects with the Pro12Pro and Pro12Ala genotypes. There was no significant difference in serum adiponectin values between subjects with the Pro12Pro and Pro12Ala genotypes (males 5.4 vs. 5.8 µg/ml, P = 0.546; females 6.9 vs. 7.2 µg/ml, P = 0.748). Adiponectin values did not differ among these two genotypes even when categorized based on their diabetes status (normal glucose tolerance Pro12Pro 7.9 vs. Pro12Ala 7.7 µg/ml, P = 0.994; diabetes Pro12Pro 4.7 vs. Pro12Ala 5.4 µg/ml, P = 0.622). Conclusion, The Pro12Ala polymorphism of the PPARG gene is not associated with serum adiponectin levels in Asian Indians. [source]

    Age-related increase in haemoglobin A1c and fasting plasma glucose is accompanied by a decrease in , cell function without change in insulin sensitivity: evidence from a cross-sectional study of hospital personnel

    DIABETIC MEDICINE, Issue 3 2002
    A. P. Yates
    Abstract Aims To examine the influence of age on glucose homeostasis in a population of healthy, non-diabetic hospital personnel. Methods One hundred and twenty female and 71 male non-diabetic individuals (fasting plasma glucose < 7.0 mmol/l) were fasted overnight prior to blood sampling. Glycated haemoglobin (HbA1c), fasting plasma glucose (FPG) and fasting plasma insulin (FPI) were measured using a BioRad Diamat automated HPLC, a Hitachi 747 analyser and a sensitive in-house radioimmunoassay, respectively. Mathematical modelling of the fasting glucose and insulin pairs (homeostasis model assessment (HOMA)) generated indices of pancreatic , cell function, HOMA-B and tissue insulin sensitivity HOMA-S. Results Spearman rank correlation analysis showed that in the whole group there was a significant negative correlation between age and HOMA-B (rs= ,0.218, P = 0.0022) and a significant positive correlation between age and both HbA1c (rs= 0.307, P = 0.0001) and FPG (rs= 0.26, P = 0.0003). There was no correlation between age and either FPI (rs= ,0.08, P = 0.266) or HOMA-S (rs= 0.024, P = 0.75). Analysis by gender showed the above associations to be present in the females (rs= ,0.243, P = 0.0076; rs= 0.304, P = 0.0007; rs= 0.32, P = 0.0004 for age vs. HOMA-B, HbA1c, and FPG, respectively). Again there was no correlation of age with FPI or insulin sensitivity. In the males there was a significant correlation of HbA1c with age (rs= 0.35, P = 0.002), but no significant correlation of age with any of the other parameters. Conclusions Glycaemic control deteriorates with age in healthy, non-diabetic individuals. Age-related rises in FPG and haemoglobin A1c result from a small but steady decline in pancreatic , cell function. Diabet. Med. 19, 254,258 (2002) [source]

    Association between oestradiol and puerperal psychosis

    ACTA PSYCHIATRICA SCANDINAVICA, Issue 2 2000
    A. Riecher-Rössler
    Objective: Postpartum psychiatric disorders with long-lasting adverse sequelae are common during the childbearing years. These disorders can be severe and resistant to conventional psychiatric treatment methods. We present two consecutive cases with puerperal psychosis who were refractory to conventional treatment methods but responded successfully to oestrogen therapy. Method: Serum oestradiol concentration was measured by radioimmunoassay and the documented oestradiol deficiency replaced with physiological oestradiol sublingually. The treatment effect was evaluated by the Brief Psychiatric Rating Scale. Results: In both cases the baseline oestradiol concentration was low (28 and 69 pmol/L). During the treatment with oestradiol, there was a concomitant elevation of the concentration of serum oestradiol, which coincided with the decline in psychotic symptoms. Conclusion: The observation of low serum oestradiol together with psychotic symptoms and successful treatment with oestradiol suggests that oestradiol may have a causal relevance to puerperal psychosis and significance in the treatment of this condition. [source]

    Effects of 4-nonylphenol on the endocrine system of the shore crab, Carcinus maenas

    ENVIRONMENTAL TOXICOLOGY, Issue 3 2008
    Christina M. Lye
    Abstract There is a considerable body of evidence to suggest that many anthropogenic chemicals, most notably xeno-estrogens, are able to disrupt the endocrine system of vertebrates. There have been few comparable studies on the effects of exposure to these chemicals that may serve as biomarkers of endocrine disruption in aquatic invertebrate species. In addition, the evidence available is complex, conflicting, and far from conclusive. The present study aimed to investigate the impact of the xeno-estrogen 4-nonylphenol (4-NP, nominal concentrations 10,100 ,g L,1) on the regulation and functioning of the endocrine system of the shore crab Carcinus maenas. It also set out to establish whether 4-NP are causing the effects (i.e., changes of exoskeletons including secondary sexual characteristics, pheromonally mediated behavior and ecdysone levels, and the presence of vt in the male hepatopancreas) found recently in wild shore crabs (Lye et al.,2005). The study utilizes morphological (e.g., gonadosomatic and hepatosomatic indices) and hormonal (ecdysteroid moulting hormone levels and the induction of female specific proteins, vitellins) biomarkers using radioimmunoassay and an indirect enzyme linked immunosorbent assay applied to the soluble protein fraction of adult male hepatopancreatic homogenates. Exposure of C. maenas to an effective concentration as low as 1.5 ,g L,1 4-NP resulted in a reduced testis weight, increased liver weight, and altered levels of ecdysone equivalents compared to controls. Induction of vitellin-like proteins was absent in all samples tested. The ecological implications and the possible mechanisms for the action of 4-NP on the response of the shore crab to xeno-estrogen exposure are discussed. © 2008 Wiley Periodicals, Inc. Environ Toxicol, 2008. [source]

    Ecdysteroid synthesis and imaginal disc development in the midge Chironomus riparius as biomarkers for endocrine effects of tributyltin

    ENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 5 2002
    Torsten Hahn
    Abstract Acute effects of the endocrine disruptor bis (tri- n -butyltin) oxide (TBTO) on molting-hormone biosynthesis and imaginaldisc development were investigated in larvae of the midge Chironomus riparius (Meigen). Ecdysteroid synthesis was measured by 24-h incubation of molting-hormone-synthesizing tissues (prothoracic glands) in vitro with or without the addition of TBTO. The amount of ecdysteroids produced was analyzed by radioimmunoassay. Developmental effects in vivo were investigated by determining the developmental phase of the genital imaginal discs before and after a 48-h exposure to TBTO in water. Sex-specific effects were found with both endpoints. Ecdysteroid synthesis was significantly reduced (analysis of variance [ANOVA], p , 0.005) in female larvae at all concentrations (TBTO-Sn at 50, 500, and 5,000 ng/L), whereas a significant elevation of the biosynthesis rate occurred in male larvae in the 500-ng/L treatment (ANOVA, p , 0.05). In vivo experiments with development of the genital imaginal disc within a 48-h exposure period revealed a significantly slower development in female larvae and a significantly faster development in male larvae (contingency tables, p , 0.001) at all concentrations tested (TBTO-Sn at 10, 50, 200, and 1,000 ng/L). These results partly coincided with the in vitro effects on molting-hormone synthesis. The 48-h median lethal concentration (LC50) was 25 ,g/L (20,30 ,g/L 95% confidence intervals). The combination of in vitro and in vivo methods has proven to be a useful approach for the detection of endocrine effects of TBTO in C. riparius at levels 2,000-fold below the LC50 value. High sensitivity and short test duration suggest that chironomids may have potential as freshwater sentinel organisms for endocrine-disrupting chemicals. [source]

    Relationship between Serum Testosterone, Dominance and Mating Success in Père David's Deer Stags

    ETHOLOGY, Issue 9 2004
    Li Chunwang
    We conducted an experiment in the Beijing Milu Park to study the social behavior of male Père David's deer, and related social behavior to social position and serum testosterone level of the stags during rut. We classified the stags into three rank classes according to their rutting behavior: ,harem master', ,challenger' and ,bachelor'. We monitored the behaviors of four ,harem masters', five ,challengers' and five ,bachelors', and analyzed serum testosterone levels in blood samples of those 14 stags using radioimmunoassay. We defined the effectiveness value, E = A/T, to assess the effectiveness of herding or mating attempts made by stags (,T' represents the frequency of herding or mating attempts made by a stag and ,A' represents the frequency of herding or mating attempts accepted by hinds). We found that: (1) the ,harem masters' and the ,challengers' displayed more frequent rut and locomotive behaviors but fewer ingestion behaviors than the ,bachelors'; (2) serum testosterone levels in the ,harem masters' and the ,challengers' were higher than that in the ,bachelors'; (3) effectiveness value of herding attempts differed significantly between the three types of stags, being highest in the ,harem masters' and the lowest in the ,bachelors'; and (4) effectiveness value of mating attempts was significantly greater for the ,harem masters' than for the ,challengers'. We conclude that: (1) reproductive behavior of the Père David's deer stags is strongly associated with social rank; (2) social roles of Père David's deer stags during the rut are related to the testosterone secretion; and (3) rank class affects the mating opportunity of the stags. [source]

    Deficits in spatial learning and synaptic plasticity induced by the rapid and competitive broad-spectrum cyclooxygenase inhibitor ibuprofen are reversed by increasing endogenous brain-derived neurotrophic factor

    EUROPEAN JOURNAL OF NEUROSCIENCE, Issue 11 2003
    Kendra N. Shaw
    Abstract Cyclooxygenase (COX), which is present in two isoforms (COX1 and 2), synthesizes prostaglandins from arachidonic acid; it plays a crucial role in inflammation in both central and peripheral tissues. Here, we describe its role in synaptic plasticity and spatial learning in vivo via an effect on brain-derived neurotrophic factor (BDNF) and prostaglandin E2 (PGE2; both measured by Elisa). We found that broad-spectrum COX inhibition (BSCI) inhibits the induction of long-term potentiation (LTP; the major contemporary model of synaptic plasticity), and causes substantial and sustained deficits in spatial learning in the watermaze. Increases in BDNF and PGE2 following spatial learning and LTP were also blocked. Importantly, 4 days of prior exercise in a running wheel increased endogenous BDNF levels sufficiently to reverse the BSCI of LTP and spatial learning, and restored a parallel increase in LTP and learning-related BDNF and PGE2. In control experiments, we found that BSCI had no effect on baseline synaptic transmission or on the nonhippocampal visible-platform task; there was no evidence of gastric ulceration from BSCI. COX2 is inhibited by glucorticoids; there was no difference in blood corticosterone levels as measured by radioimmunoassay in any condition. Thus, COX plays a previously undescribed, permissive role in synaptic plasticity and spatial learning via a BDNF-associated mechanism. [source]

    Abnormal substance P release from the spinal cord following injury to primary sensory neurons

    EUROPEAN JOURNAL OF NEUROSCIENCE, Issue 1 2000
    Marzia Malcangio
    Abstract The neuropeptide substance P (SP) modulates nociceptive transmission within the spinal cord. Normally, SP is uniquely contained in a subpopulation of small-calibre axons (A,- and C-fibres) within primary afferent nerve. However, it has been shown that after nerve transection, besides being downregulated in small axons, SP is expressed de novo in large myelinated A,-fibres. In this study we investigated whether, following peripheral nerve injury, SP was released de novo from the spinal cord after selective activation of A,-fibres. Spinal cords with dorsal roots attached were isolated in vitro from rats 2 weeks following distal sciatic axotomy or proximal spinal nerve lesion (SNL). The ipsilateral dorsal roots were electrically stimulated for two consecutive periods at low- or high-threshold fibre strength, spinal cord superfusates were collected and SP content was determined by radioimmunoassay. SNL, but not axotomized or control rat cords, released significant amounts of SP after selective activation of A,-fibres. Not only do these data support the idea that A, myelinated fibres contribute to neuropathic pain by releasing SP, they also illustrate the importance of the proximity of the lesion to the cell body. [source]

    Effect of parathyroid hormone-related protein on fibroblast proliferation and collagen metabolism in human skin

    EXPERIMENTAL DERMATOLOGY, Issue 4 2002
    Emanuela Maioli
    Abstract: The parathyroid hormone-related protein (PTHrp), structurally similar to the parathyroid hormone (PTH) in its NH2 -terminal part, was first identified as a tumour-derived peptide responsible for a paraneoplastic syndrome known as humoral hypercalcemia of malignancy. The PTHrp gene is expressed not only in cancer but also in normal tissues during adult and/or fetal life, where it plays predominantly paracrine and/or autocrine roles. In the skin PTHrp produced by keratinocytes acts on fibroblasts by complex cooperative circuits involving cytokines and growth factors. In this report, we studied the direct effects of synthetic PTHrp 1,40 on proliferation and collagen synthesis and matrix metalloproteinase-2 (MMP-2) activity in cultures of fibroblasts isolated from normal human skin. Fibroblasts exposure to varying doses of PTHrp for 48 h, significantly and dose-dependently inhibited proliferation evaluated by [3H]-thymidine incorporation into DNA. A dose-dependent stimulation of cAMP released into the medium was concomitantly observed. In contrast, PTHrp had no effect on collagen synthesis evaluated either by [3H]-proline incorporation or by radioimmunoassay (RIA) of the carboxyterminal fragment of type I procollagen (PICP). MMP-2 activity, evaluated by quantitative zymographic analysis, was significantly increased by PTHrp treatment at doses of 160 and 320 nM. These findings indicate that PTHrp may play a role in normal dermal physiology by controlling both fibroblast proliferation and extracellular matrix degradation. [source]

    Presence of immunoreactive ,-endorphin in human skin

    EXPERIMENTAL DERMATOLOGY, Issue 5 2001
    M. Wintzen
    Abstract: The production and its induction by ultraviolet radiation (UVR) of proopiomelanocortin (POMC)-derived peptides by keratinocytes has been reported, albeit not consistently. Recently we demonstrated that only under specific culturing conditions human keratinocytes are capable of producing a ,-endorphin (,E)-like peptide with the characteristics of ,-lipotropin (,LPH). Here the presence and UV-induction of ,E-immunoreactivity (,E-IR) in keratinocytes in human skin in vivo was investigated. ,E-IR was detectable by immunohistochemistry in keratinocytes of the follicular matrix and to some extent in cells of sweat ducts, but was absent from epidermal keratinocytes. Absence of ,E-IR was confirmed by radioimmunoassay of HPLC-fractionated extracts of normal epidermis. Repeated exposure to solar-simulated UVR had no effect. This investigation is the first to demonstrate the presence of ,E-immunoreactive material in the follicular matrix of corporal hairs and in duct cells of sweat glands. The possible meaning of these results is discussed. [source]

    Testosterone response to GnRH in a female songbird varies with stage of reproduction: implications for adult behaviour and maternal effects

    FUNCTIONAL ECOLOGY, Issue 4 2007
    JODIE M. JAWOR
    Summary 1Despite considerable recent interest in plasma and yolk testosterone (T) in female birds, relatively little is known about environmental regulation of female T, individual variation in female T or the relationship between plasma and yolk T. 2In breeding females of a wild population of dark-eyed junco (Junco hyemalis), we assessed variation in the responsiveness of the hypothalamo-pituitary-gonadal (HPG) axis to a challenge with gonadotropin-releasing hormone (GnRH) by measuring circulating T before and 30 min after a standardized injection of GnRH. We asked whether response to challenge varied seasonally or with stage of reproduction and whether it was repeatable within individuals or related to T deposited in eggs. 3Initial and post-challenge levels of T were measured using enzyme immunoassay. In a subset of these females, luteinising hormone (LH) was measured using radioimmunoassay (RIA). In addition, eggs were collected from nests of 15 females that had received a GnRH challenge, and yolk T was measured using RIA. 4During most of the breeding season, plasma T did not increase in response to GnRH. GnRH consistently caused increases in plasma T only during the 7 days before oviposition, when females were rapidly depositing yolk in eggs but had not yet begun to lay them. Among a small subset of females we found a positive correlation between the magnitude of this increase in plasma T in response to GnRH during egg development and the amount of T deposited in the yolk of eggs collected at a later time. 5These results suggest that ovarian response to GnRH-induced increases in LH is greatest when females are actively depositing yolk into eggs. Factors that stimulate the release of GnRH during egg formation may result in higher levels of plasma T which could influence adult female behaviour. Further, because plasma T was correlated with later yolk T, factors that stimulate GnRH release may also lead to higher levels of yolk T potentially influencing offspring development or behaviour. [source]

    Neuron,Glia Signaling in Trigeminal Ganglion: Implications for Migraine Pathology

    HEADACHE, Issue 7 2007
    Srikanth Thalakoti BS
    Objective.,The goal of this study was to investigate neuronal,glial cell signaling in trigeminal ganglia under basal and inflammatory conditions using an in vivo model of trigeminal nerve activation. Background.,Activation of trigeminal ganglion nerves and release of calcitonin gene-related peptide (CGRP) are implicated in the pathology of migraine. Cell bodies of trigeminal neurons reside in the ganglion in close association with glial cells. Neuron,glia interactions are involved in all stages of inflammation and pain associated with several central nervous system (CNS) diseases. However, the role of neuron,glia interactions within the trigeminal ganglion under normal and inflammatory conditions is not known. Methods.,Sprague,Dawley rats were utilized to study neuron,glia signaling in the trigeminal ganglion. Initially, True Blue was used as a retrograde tracer to localize neuronal cell bodies in the ganglion by fluorescent microscopy and multiple image alignment. Dye-coupling studies were conducted under basal conditions and in response to capsaicin injection into the TMJ capsule. S100B and p38 expression in neurons and glia were determined by immunohistochemistry following chemical stimulation. CGRP levels in the ganglion were measured by radioimmunoassay in response to capsaicin. In addition, the effect of CGRP on the release of 19 different cytokines from cultured glial cells was investigated by protein microarray analysis. Results.,In unstimulated control animals, True Blue was detected primarily in neuronal cell bodies localized in clusters within the ganglion corresponding to the V3 region (TMJ capsule), V2 region (whisker pad), or V1 region (eyebrow and eye). However, True Blue was detected in both neuronal cell bodies and adjacent glia in the V3 region of the ganglion obtained from animals injected with capsaicin. Dye movement into the surrounding glia correlated with the time after capsaicin injection. Chemical stimulation of V3 trigeminal nerves was found to increase the expression of the inflammatory proteins S100B and p38 in both neurons and glia within the V3 region. Unexpectedly, increased levels of these proteins were also observed in the V2 and V1 regions of the ganglion. CGRP and the vesicle docking protein SNAP-25 were colocalized in many neuronal cell bodies and processes. Decreased CGRP levels in the ganglion were observed 2 hours following capsaicin stimulation. Using protein microarray analysis, CGRP was shown to differentially regulate cytokine secretion from cultured trigeminal ganglion glia. Conclusions.,We demonstrated that activation of trigeminal neurons leads to changes in adjacent glia that involve communication through gap junctions and paracrine signaling. This is the first evidence, to our knowledge, of neuron,glia signaling via gap junctions within the trigeminal ganglion. Based on our findings, it is likely that neuronal,glial communication via gap junctions and paracrine signaling are involved in the development of peripheral sensitization within the trigeminal ganglion and, thus, are likely to play an important role in the initiation of migraine. Furthermore, we propose that propagation of inflammatory signals within the ganglion may help to explain commonly reported symptoms of comorbid conditions associated with migraine. [source]

    Dipeptidyl peptidase expression during experimental colitis in mice

    INFLAMMATORY BOWEL DISEASES, Issue 8 2010
    Roger Yazbeck PhD
    Abstract Background: We have previously demonstrated that inhibition of dipeptidyl peptidase (DP) activity partially attenuates dextran sulfate sodium (DSS) colitis in mice. The aim of this study was to further investigate the mechanisms of this protection. Materials and Methods: Wildtype (WT) and DPIV,/, mice consumed 2% DSS in drinking water for 6 days to induce colitis. Mice were treated with saline or the DP inhibitors Ile-Pyrr-(2-CN)*TFA or Ile-Thia. DP mRNA and enzyme levels were measured in the colon. Glucagon-like peptide (GLP)-2 and GLP-1 concentrations were determined by radioimmunoassay, regulatory T-cells (Tregs) by fluorescence activated cell sorting (FACS) on FOXp3+T cells in blood, and neutrophil infiltration assessed by myeloperoxidase (MPO) assay. Results: DP8 and DP2 mRNA levels were increased (P < 0.05) in WT+saline mice compared to untreated WT mice with colitis. Cytoplasmic DP enzyme activity was increased (P < 0.05) in DPIV,/, mice at day 6 of DSS, while DP2 activity was increased (P < 0.05) in WT mice with colitis. GLP-1 (63%) and GLP-2 (50%) concentrations increased in WT+Ile-Pyrr-(2-CN)*TFA mice compared to day-0 controls. MPO activity was lower in WT+Ile-Thia and WT+Ile-Pyrr-(2-CN)*TFA treated mice compared to WT+saline (P < 0.001) at day 6 colitis. Conclusions: DP expression and activity are differentially regulated during DSS colitis, suggesting a pathophysiological role for these enzymes in human inflammatory bowel disease (IBD). DP inhibitors impaired neutrophil recruitment and maintenance of the Treg population during DSS-colitis, providing further preclinical evidence for the potential therapeutic use of these inhibitors in IBD. Finally, DPIV appears to play a critical role in mediating the protective effect of DP inhibitors. Inflamm Bowel Dis 2010 [source]

    Dental pulp fibroblasts express neuropeptide Y Y1 receptor but not neuropeptide Y

    INTERNATIONAL ENDODONTIC JOURNAL, Issue 10 2010
    S. A. Killough
    Killough SA, Lundy FT, Irwin CR. Dental pulp fibroblasts express neuropeptide Y Y1 receptor but not neuropeptide Y. International Endodontic Journal, 43, 835,842, 2010. Abstract Aim, To investigate whether dental pulp fibroblasts express neuropeptide Y (NPY) and NPY-Y1 in vitro and to determine the effects of the cytokines including interlukin-1, (IL-1,), TGF- ,1, substance P and NPY on the expression of NPY Y1. Methodology, Three primary fibroblast cell strains were obtained from freshly extracted human third molar teeth. RT-PCR was utilized to detect expression of NPY and mRNA expression. Membrane protein samples were isolated, and protein expression was determined by Western blotting. Radioimmunoassay was used to quantify NPY expression in healthy (n = 35) and carious (n = 39) whole pulp samples, and the student's t -test was used to test for statistical significance. In addition, the 3-(4,5-Dimethylthiazol,2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to assay fibroblast cell growth. Results, mRNA transcripts were found in all three fibroblast cell populations with the cytokines having a stimulatory effect on its expression (P < 0.05). NPY mRNA was not detected in the cell strains. NPY-Y1 receptor protein expression was visualized by Western blotting, and there was no effect of IL-1, or TGF- ,1 on its expression. The mean concentration of NPY-Ir determined by radioimmunoassay in non-carious teeth was 19.40 ng g,1 (±17.03 SD) compared to 29.95 ng g,1 (±20.99 SD) in carious teeth (P < 0.05). Conclusion, Human dental pulp fibroblasts express, but do not synthesize, NPY, demonstrating that the fibroblast is a target cell for NPY. The effect of proinflammatory cytokines suggests that fibroblasts play a neuroimmunomodulatory role in the pulpal response to dental caries and injury. [source]

    Estradiol levels in prepubertal boys and girls , analytical challenges

    INTERNATIONAL JOURNAL OF ANDROLOGY, Issue 5 2004
    Katrine Bay
    Summary Increasing evidence points at an important function of low concentrations of estradiol (E2) in prepubertal boys and girls. E2 serum levels in prepubertal children are, however, often immeasurable in conventional E2 assays. This strongly hampers further investigation of the physiological relevance of E2 in children. In addition, there is an increasing concern of the potential effect of exposure to endocrine disrupters with estrogenic or antiandrogenic activity on pubertal development. A requirement of assessing the instance for this concern, adds further to the demands for applicable methodologies for the evaluation of the sensitivity of the organism to low E2 concentrations. Traditionally, E2 is measured by use of the radioimmunoassay (RIA). As an ultrasensitive alternative to the RIA, a recombinant cell bioassay has been developed. In this review, methodological aspects for these methods of analysis are examined and their applicability for evaluation of low E2 serum concentrations in children is estimated. Furthermore, available data on E2 levels in prepubertal boys and girls are evaluated and discussed, taking into consideration the limitations of the methods of analysis. In conclusion, there is a pronounced demand for new and improved methods of analysis for accurate and sensitive evaluation of low concentrations of E2. [source]

    Effects of steroids on oxytocin secretion by the human prostate in vitro

    INTERNATIONAL JOURNAL OF ANDROLOGY, Issue 1 2004
    S. J. Assinder
    Summary Oxytocin (OT) concentrations are elevated in prostate tissue of patients with benign prostatic hyperplasia (BPH). Oxytocin specifically increases growth, 5 , -reductase activity and contractility in the prostate. In the rat prostatic OT concentrations are regulated by gonadal steroids, with androgens reducing but oestrogens increasing OT concentrations. The regulation of prostatic oxytocin in man is not understood. This study investigates the effects of gonadal steroids on oxytocin production by the human prostate. Primary explants (approx. 1 mm3) of prostate tissue from patients with BPH were incubated in Dulbecco's modified Eagle's media in the absence or presence of 10 nmol/L testosterone (T), 10 nmol/L dihydrotestosterone (DHT), T or DHT plus 100 nmol/L of the anti-androgen cyproterone acetate (CPA), 55 pmol/L diethylstilbestrol (DES), or DES plus DHT. The amount of oxytocin secreted into the media after 3 days was measured by radioimmunoassay. Testosterone and DHT significantly increased oxytocin concentrations secreted into the media from 0.86 ± 0.11 ng/g of tissue (control) to 1.51 ± 0.14 ng/g (p < 0.01) and 1.54 ± 0.13 ng/g (p < 0.05), respectively. Incubation of tissue samples with CPA resulted in oxytocin concentrations similar to control levels. Treatment with DES caused a significant increase from 1.99 ± 0.71 to 3.98 ± 1.36 ng/g (p < 0.05). A similar increase was measured in media of tissue incubated in DES plus DHT (p < 0.001). The results demonstrate that, unlike the rat where androgens decrease oxytocin, in hyperplastic human prostate tissue both androgens and oestrogens increase oxytocin. This imbalance in the regulation of oxytocin may result in promoting prostatic overgrowth in the pathogenesis of BPH. [source]