1,303 research outputs found
Development of a protein binding assay for teleost insulin like growth factor (IGF)-like : relationships between growth hormone (GH) and IGF-like in the blood of rainbow trout
No full textUsing rainbow trout plasma protein (IGF-BP) which specifically binds human insulin-like growth factor (IGF) (Niu and Le Bail 1993), we have developed an assay to measure plasma IGF-like levels in different teleost species. Before the assay and to prevent interference by IGF-BP, IGF-like was extracted from all samples, using SP Sephadex C-25 in acidic conditions. After this treatment, contamination of the IGF fraction by IGF-BP which was estimated by binding assay, was approximately 5%, and was not detectable by western ligand blot.Human IGF-I was used as standard and labelled hormone. Sensitivity of the assay was 0.15-0.40 ng/ml (ED90) and ED50 was 1-3 ng/ml. hIGF-II crossreaction was partial and no significant displacement was observed with Insulin from different species or with other hormones. Inhibition curves were obtained with plasma IGF fractions (but not with tissue extracts) from teleost and mammals and are parallel to the standard curve. These results suggest that the protein binding assay can quantify an IGF-like factor in the plasma of teleost and that the binding sites of IGF are well conserved during vertebrate evolution.Using this IGF binding assay, IGF-like was measured in parallel with growth hormone (GH) in plasma from young rainbow trout killed every 1.5h throughout one day. The daily profiles for both hormones, which appear pulsatile, are similar. A significant correlation was observed between GH levels and IGF-like levels with a 1.5h delay. Analogous observations were obtained in individual catheterized adult rainbow trout. Although plasma GH levels differ greatly between fish, less variability is found with IGF-like. In a third experiment, rainbow trout were starved or submitted to bovine GH treatment for four weeks. Starved fish, in which plasma GH levels increased, had plasma IGF-like level significantly lower than in fed fish. In bGH injected fish, plasma IGF-like level was significantly higher than in non-injected fish. These results suggest that, as in mammals, IGF-like secretion depends on plasma GH level and could be modulated by the nutritional status of fish.Un dosage des somatomédines (IGF) plasmatiques de téléostéens a été mise au point, en utilisant une protéine du sérum de truite qui lie spécifiquement les IGF humains (Niu and Le Bail 1993). Pour éliminer les risques d'interférence dus aux protéines de liaison (IGF-BP), I'activité IGF des différents échantillons a été extraite à l'aide de SP Sephadex C-25 en condition acide. La contamination en IGF-BP de ces extraits est estimé à 5% par dosage de la liaison et n'est pas détectable en western ligand blot. L'IGF-I humain a ete utilisée comme standard et comme traceur. La sensibilité du dosage est de 0.15-0.40 ng/ml (ED90) et 1'ED50 varie entre I et 3 ng/ml. L'IGF-II humain est reconnue partiellement mais aucune réaction croisée n'est observée avec de l'insuline de différentes espèces ni avec les autres hormones testées. Les courbes d'inhibition obtenues avec les sera de mammifères et de téléostéens sont parallèles a la courbe standard. Ces résultats montrent que le dosage par protéine de liaison est capable de quantifier une activité de type IGF dans le sérum des téléostéen, et que le site de liaison des IGF est resté bien conservé au cours de l'évolution des vertébrés. En utilisant ce dosage, nous avons mesure l'activité IGF et les niveaux d'hormone de croissance (GH) dans des plasmas de jeunes truites arc-en-ciel abattues toutes les heures et demie durant 24h. Les profils nycthéméraux des deux hormones, qui sont de type pulsatile, apparaissent similaires. Une corrélation significative est observée entre les niveaux de GH et les activités IGF circulant une heure et demie plus tard. Des observations analogues ont été faites chez des truites adultes cathétérisées. Cependant, les niveaux plasmatiques de GH sont très différents d'un animal à l'autre, alors que les variations de l'activité IGF sont moins prononcées. Dans une troisième expérience, des truites ont été réparties en trois groupes: un groupe contrôle, un groupe traité avec de la GH bovine et un groupe soumis a un jeûne prolongé. Chez les animaux a jeun, les niveaux de GH augmentent alors que les activités IGF diminuent. Chez les animaux injectés avec de la GH, les activités IGF sont significativement plus levées que chez les animaux témoins. Ces résultats suggèrent que, comme chez les mammifères, la sécrétion des IGF plasmatiques est contrôlé par les niveaux circulants de GH et que les variations de la réceptivité tissulaire la GH dépendent de l'état nutritionnel des animaux
Short term growth hormone (GH) treatment of GH-deficient adults increases body sodium and extracellular water, but not blood pressure
No full textInitiation of GH treatment in adults is frequently complicated by the development of symptomatic fluid retention. To investigate the mechanism and extent of fluid retention that occurs with dosages of GH used in the treatment of GH-deficient adults, we conducted a double blind study in which seven GH-deficient patients (aged 24-74 yr) each received in random order daily sc injections of placebo, a physiological dose of GH (0.04 U/kg, low dose), and a supraphysiological dose of GH (0.08 U/kg, high dose) for 7 days, separated by 21-day washout periods. On the seventh day, measurements were made of serum insulin-like growth factor I, body weight, exchangeable sodium, plasma volume, angiotensinogen, PRA, aldosterone, atrial natriuretic peptide (ANP), and mean 24-h ambulatory heart rate and blood pressure. GH significantly increased mean insulin-like growth factor I levels from 105 ± 11 to 304 ± 45 μg/L during low dose treatment (P = 0.006) and 400 ± 76 μg/L during high dose treatment (P = 0.004). High dose GH caused a 1.2 ± 0.3 kg increase in body weight (P = 0.01) and a 193 ± 65 mmol increase in exchangeable sodium (P = 0.008). Low dose GH had a lesser effect, with no significant increase in body weight, but an increase in exchangeable sodium of 113 ± 37 mmol (P = 0.02). Plasma volume was not significantly affected by GH treatment. Mean supine angiotensinogen levels were significantly higher during both GH treatments compared to placebo (low dose, P = 0.017; high dose, P = 0.028) as were mean supine pRA levels (low dose, P = 0.0002; high dose, P = 0.0025). Supine angiotensin II, aldosterone, and ANP levels were not significantly affected by GH treatment. There was no significant change from placebo in any of the sodium-regulating hormones in the erect posture. The mean 24-h heart rate was significantly higher during low dose (82 ± 2 beats/min; P = 0.0001) and high dose (88 ± 3 beats/min; P = 0.0001) GH treatment than during placebo (67 ± 3 beats/min). However, no significant change in mean 24-h systolic or diastolic blood pressure was observed. In summary, acute GH administration using doses currently employed in treating adults causes a dose-related increase in body weight and body sodium, but no associated increase in blood pressure. We conclude that 1) sodium retention is a physiological effect of GH, but does not cause an acute rise in blood pressure; and 2) the mechanism of sodium and fluid retention is not primarily due to enhanced aldosterone secretion or inhibition of ANP release, but more likely to a direct renal tubular effect
Etude du polymorphisme de l'hormone de croissance (GH) chez la truite arc-en-ciel (Oncorhynchus mykiss): Mise en évidence de différentes formes de la GH. Mise au point d'un test biologique in vitro de la GH. Recherche d'une protéine de liaison sérique liant la GH
No full textThe aim of in this work was to reveal the different forms of GH in the pituitary and serum of rainbow trout (Oncorhynchus mykiss) and together with their biochemical and biological characteristics.Nous nous étions fixés, comme but de ce travail effectué chez la truite arc-en-ciel (Oncorhynchus mykiss), la mise en évidence des différentes formes de la GH hypophysaire et sérique et d'en connaître les caractéristiques biochimiques et biologiques
RNAseq analysis of fast skeletal muscle in restriction-fed transgenic coho salmon (Oncorhynchus kisutch) : an experimental model uncoupling the growth hormone and nutritional signals regulating growth
Get PDFBackground Coho salmon (Oncorhynchus kisutch) transgenic for growth hormone (Gh) express Gh in multiple tissues which results in increased appetite and continuous high growth with satiation feeding. Restricting Gh-transgenics to the same lower ration (TR) as wild-type fish (WT) results in similar growth, but with the recruitment of fewer, larger diameter, muscle skeletal fibres to reach a given body size. In order to better understand the genetic mechanisms behind these different patterns of muscle growth and to investigate how the decoupling of Gh and nutritional signals affects gene regulation we used RNA-seq to compare the fast skeletal muscle transcriptome in TR and WT coho salmon. Results Illumina sequencing of individually barcoded libraries from 6 WT and 6 TR coho salmon yielded 704,550,985 paired end reads which were used to construct 323,115 contigs containing 19,093 unique genes of which >10,000 contained >90 % of the coding sequence. Transcripts coding for 31 genes required for myoblast fusion were identified with 22 significantly downregulated in TR relative to WT fish, including 10 (vaspa, cdh15, graf1, crk, crkl, dock1, trio, plekho1a, cdc42a and dock5) associated with signaling through the cell surface protein cadherin. Nineteen out of 44 (43 %) translation initiation factors and 14 of 47 (30 %) protein chaperones were upregulated in TR relative to WT fish. Conclusions TR coho salmon showed increased growth hormone transcripts and gene expression associated with protein synthesis and folding than WT fish even though net rates of protein accretion were similar. The uncoupling of Gh and amino acid signals likely results in additional costs of transcription associated with protein turnover in TR fish. The predicted reduction in the ionic costs of homeostasis in TR fish associated with increased fibre size were shown to involve multiple pathways regulating myotube fusion, particularly cadherin signaling.Peer reviewe
GH and the cardiovascular system: an update on a topic at heart
Get PDFIn this review, the importance of growth hormone (GH) for the maintenance of normal cardiac function in adult life is discussed. Physiological effects of GH and underlying mechanisms for interactions between GH and insulin-like growth factor I (IGF-I) and the cardiovascular system are covered as well as the cardiac dysfunction caused both by GH excess (acromegaly) and by GH deficiency in adult hypopituitary patients. In both acromegaly and adult GH deficiency, there is also increased cardiovascular morbidity and mortality possibly linked to aberrations in GH status. Finally, the status of the GH/IGF-I system in relation to heart failure and the potential of GH as a therapeutic tool in the treatment of heart failure are reviewed in this article. © 2014 The Author(s)
Avaliação da expressão imuno-histoquímica das células retais produtoras de grelina em ratos Wistar submetidos à dieta de cafeteria
Get PDFDissertação (mestrado) - Universidade Federal de Santa Catarina, Centro de Ciências da Saúde, Programa de Pós-Graduação em Ciências Médicas, Florianópolis, 2012Introdução: Na atualidade o sobrepeso e a obesidade, induzidos pela dieta e pelo sedentarismo, são doenças muito prevalentes em vários países. Considerados uma epidemia mundial pela Organização Mundial da Saúde (OMS), são importantes fatores de risco para inúmeras afecções, entre elas as neoplasias colorretais. Estudos prévios sugerem que em portadores de obesidade ou sobrepeso há aumento da incidência desse tipo de neoplasia e a piora do prognóstico. Com a descoberta da grelina, em 1999, que é um hormônio orexígeno com potente ação sobre a liberação do hormônio do crescimento (GH) e atuação em diversos sistemas orgânicos, fisiológicos e na carcinogênese, desenvolveram-se diversas pesquisas para elucidar o seu papel e sua importância em todas essas funções. Recentemente, evidenciou-se aumento da expressão de grelina em amostras teciduais de neoplasias colorretais quando comparadas à tecidos colônicos normais. Objetivos: Investigar o impacto da dieta de cafeteria na expressão imuno-histoquímica retal da grelina e avaliar o tipo morfológico das células identificadas. Desenho do Estudo: Estudo analítico experimental do tipo corte transversal com intervenção. Método: Vinte e quatro ratos Wistar machos foram distribuídos em 4 subgrupos de 6 animais cada denominados: SGR1 (ração e água) e SGC1 (dieta de cafeteria, ração e água) por um período de 30 dias; SGR2 (ração e água) e SGC2 (dieta de cafeteria, ração e água) por um período de 60 dias. O peso dos animais e do reto amputado, o número e o tipo de células imunorreativas à grelina foram registrados e comparados entre os subgrupos. Na análise estatística foram utilizados os testes ANOVA com correção de Bonferroni, qui quadrado, teste t pareado e teste t de Student. Resultados: Houve aumento significativo do peso corporal em todos os subgrupos (P 0,001). O peso no dia da cirurgia foi significativamente maior na comparação do SGC2 com os demais subgrupos: SGR1 (P=0,003), SGC1 (P= 0,010) e SGR2 (P= 0,001). Não houve diferença na comparação da média do número absoluto de células imunorreativas (P= 0,685) e na comparação entre expressão e não expressão (P=0,330) entre os subgrupos, nem entre o peso final (P=0,993) e o peso retal (P= 0,230) com a imunoexpressão. Todas as células imunorreativas identificadas foram do tipo "fechado". Conclusão: Os resultados do presente estudo não demonstraram influência da dieta de cafeteria sobre o número de células retais imunorreativas à grelina quando comparado aos controles nem aos dois períodos de ingesta e apenas células imunorreatoras do tipo "fechado" foram identificadas no reto de ratos Wistar.Abstract : Introduction: Obesity and overweight induced by diet and lifestyle are considered a global epidemic by the World Health Organization (WHO), especially in western countries. They are important risk factors for several conditions, including colorectal cancer. Obese and overweight patients have already demonstrated an increased incidence and worse prognosis associated with colorectal cancer. Since discovery of ghrelin in 1999, which is a potent orexigenic hormone and releaser of the growth hormone (GH) that has many physiologic functions, several studies began to elucidate its role and importance in several areas including carcinogenesis. Recently it was demonstrated an increase on tissue ghrelin expression in colorectal cancers samples. Objectives: To investigate the impact of cafeteria diet on ghrelin expression in rectal tissue and identify the morphologic cell type. Study Design: Analytical and experimental transversal study with intervention. Methods: Twenty-four male Wistar rats were divided into 4 subgroups of 6 animals each: RCG1 (rat chow and water) and CAFG1 (cafeteria diet, rat chow and water) for a period of 30 days; RCG2 (rat chow and water) and CAFG2 (cafeteria diet, rat chow and water) for a period of 60 days. The animal and rectal weight, the number and the type of immunoreactive ghrelin cells were recorded and compared between the subgroups. ANOVA with Bonferroni correction, chi square, t student and paired t tests were applied. Results: There was a significant increase in body weight in all subgroups (P=0,001). The weight on the operation day was significantly higher in the CAFG2 in comparison with others subgroups: RCG1 (P=0,003), CAFG1 (P=0,010) and RCG2 (P=0,001). There was no difference in the total of immunoreactive cells (P=0,685), presence or absence of ghrelin expression between the subgroups (P=0, 330), nor in the final weight (P= 0,993) and rectal weight (P=0,230) in comparison with imunoexpression. All the immunoreactive cells identified were closed-type. Conclusion: The present study showed no influence of cafeteria diet on the amount of immunoreactive rectal cells of ghrelin when compared to controls and with two periods of exposure. Only one type (closed-type) of immunoreactive cells was expressed in the rectum
Book Review: Comrades Betrayed: Jewish World War I Veterans Under Hitler
Get PDFThis is a pre-copyedited, author-produced version of an article accepted for publication in [German History] following peer review. The version of record [Grady, T. (2021). [Review of the book Comrades Betrayed: Jewish World War I Veterans Under Hitler by M. Geheran]. German History, 39(3), 478–479] is available online at: https://academic.oup.com/gh/article/39/3/478/6308748Book review of Comrades Betrayed: Jewish World War I Veterans Under HitlerUnfundedAAM out of embargo 24/06/2023, output uploaded to CR 30/01/202
Espressione genica indotta dall'ormone somatotropo nei monociti di bambini sani e con deficit di GH
Get PDF2018 - 2019Somatotropic hormone (GH) has transcriptional effects on the cells of many organs, directly by activating its receptor (GHR) or indirectly through induction of IGF-1 or other mediators. The presence of GHR in almost all cellular tissues makes GH action systemic even if, to date, still not well characterized. The immune system is among the districts where the effect of the somatotropic hormone is documented by mechanisms that are still poorly understood.The primary objective of this study is to determine the transcriptional effect of GH on peripheral blood monocytes. These cells were chosen for the significant expression of GHR on their surface and because they are easily accessible. Although the transcriptional response to somatotropic hormone is specific tissue, the study of the effects of GH on monocytes can serve as a model for other cell types and highlight differences between healthy subjects and those with GH deficiency (GHD).The diagnosis of GHD, during the developmental age, is classically based on the clinical evaluation associated with radiological and laboratory investigations (GH-IGF-1 axis stimulus test). Although provocation tests represent diagnostic gold standard, they have poor reproducibility and accuracy and are characterized by a considerable number of false positives and sometimes negatives.The secondary objective of this study is to identify differential transcriptional profiles between healthy subjects and with GHD.For this purpose, the gene expression of monocytes from healthy children and with GHD was compared in culture, under basal conditions and after stimulation with recombinant GH (rh-GH).Two groups of 12 subjects were selected, group S: healthy male children with normal height and growth rate and group D: children of the same sex and age and suffering from GHD, not yet in replacement therapy. Peripheral blood monocytes were purified by subtraction with monoclonal antibodies and the purity level was determined by laminar flow cytofluorimetry with monoclonal antibodies. Monocytes were grown for 24 hours with and without rh-GH. Total RNA was extracted and frozen until the analysis was performed simultaneously for all the experimental points using the Next Generation Sequencing methodology on Illumina platform. Differential expression of mRNA was analyzed by comparing the monocytes of healthy children and with GHD, stimulated in culture with rh-GH or not stimulated: GHD not stimulated (D-CNTR) vs healthy not stimulated (S-CNTR); healthy non-stimulated (S-CNTR) vs healthy stimulated (S-GH); non-stimulated GHD (D-CNTR) vs stimulated GHD (D-GH); GHD stimulated (D-GH) vs healthy stimulated (S-GH).The analysis between D-CNTR vs S-CNTR groups identified 58 genes with differential expression. Furthermore, 23 genes were modulated by GH in healthy children and 4 genes in children with GHD. Differential analysis between D-GH vs S-GH groups, on the other hand, identified 150 genes with differential expression.Finally, analysis performed by Ingenuity Pathway Analysis software showed a significant increase in NFAT immune pathways and dendritic cell maturation and a consistent increase in the expression of dendritic markers (HLA-A, HLA-C, CCR7) in monocytes of children with GHD compared to healthy children, after stimulation in culture with recombinant GH.In conclusion, the results of this study have demonstrated a clear transcriptional effect of GH on monocytes, direct and indirect through intermediate mediators, suggesting to evaluate the pro-inflammatory status of children with growth hormone deficiency more in depth.Furthermore, this study identified a gene expression profile of monocytes in children with GHD which, once verified in a larger number of patients, could represent an alternative to stimulus tests and guide the diagnosis of GH deficiency.Finally, our study opens future perspectives in order to identify a transcriptional profile or specific genes specific to GHD condition. [edited by Author]XXXII cicl
Comparison of Two Methods for In Vivo Estimation of the Glenohumeral Joint Rotation Center (GH-JRC) of the Patients with Shoulder Hemiarthroplasty
Get PDFDetermination of an accurate glenohumeral-joint rotation center (GH-JRC) from marker data is essential for kinematic and dynamic analysis of shoulder motions. Previous studies have focused on the evaluation of the different functional methods for the estimation of the GH-JRC for healthy subjects. The goal of this paper is to compare two widely used functional methods, namely the instantaneous helical axis (IHA) and symmetrical center of rotation (SCoRE) methods, for estimating the GH-JRC in vivo for patients with implanted shoulder hemiarthroplasty. The motion data of five patients were recorded while performing three different dynamic motions (circumduction, abduction, and forward flexion). The GH-JRC was determined using the CT-images of the subjects (geometric GH-JRC) and was also estimated using the two IHA and SCoRE methods. The rotation centers determined using the IHA and SCoRE methods were on average 1.4760.62 cm and 2.0760.55 cm away from geometric GH-JRC, respectively. The two methods differed significantly (two-tailed p-value from paired t-Test ,0.02, post-hoc power ,0.30). The SCoRE method showed a significant lower (two-tailed p-value from paired t-Test ,0.03, post-hoc power ,0.68) repeatability error calculated between the different trials of each motion and each subject and averaged across all measured subjects (0.6260.10 cm for IHA vs. 0.4360.12 cm for SCoRE). It is concluded that the SCoRE appeared to be a more repeatable method whereas the IHA method resulted in a more accurate estimation of the GH-JRC for patients with endoprostheses.Biomechanical EngineeringMechanical, Maritime and Materials Engineerin
CRM197-conjugated peptides vaccine of HCMV pp65 and gH induce maturation of DC and effective viral-specific T cell responses
No full textHuman cytomegalovirus (HCMV) infection is prevalent worldwide, and there is currently no licenced HCMV vaccine to control it. Therefore, developing an effective HCMV vaccine is a significant priority. Because of their excellent immunogenicity, the crucial components of HCMV, phosphoprotein 65 (pp65) and glycoproteins H (gH) are potential target proteins for HCMV vaccine design. In this study, we predicted and screened the dominant antigenic epitopes of B and T cells from pp65 and gH conjugated with the carrier protein cross-reacting material 197 (CRM197) to form three peptide-CRM197 vaccines (pp65-CRM197, gH-CRM197, and pp65-CRM197+gH-CRM197). Furthermore, the immunogenicity of the peptide-CRM197 vaccines and their effects on dendritic cells (DCs) were explored. The results showed that three peptide-CRM197 vaccines could induce maturation of DCs through the p38 MAPK signalling pathway and promote the release of proinflammatory factors, such as TNF-α and interleukin (IL) −6. Meanwhile, the peptide-CRM197 vaccines could effectively activate T cell and humoral immunity, which were far better than the inactivated HCMV vaccine. In conclusion, we constructed three peptide-CRM197 vaccines, which could induce multiple immune effects, providing a novel approach for HCMV vaccine design
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