Insulin-like Growth Research Articles

Growth, rumen fermentation and plasma metabolites of Holstein male calves fed fermented corn gluten meal during the postweaning stage

The objective of this experiment was to determine whether the dietary inclusion of fermented corn gluten meal (FCGM) would alter the growth, rumen fermentation and plasma metabolites of postweaning calves. Twenty-four Holstein male calves (mean ± SEM: ages = 56.21 ± 2.43 d; body weight = 83.36 ± 3.62 kg) were kept in individual pens (2.0 × 2.5 m2). After 2 weeks of adaptation, the calves were blocked for body weight and age before being randomly assigned to 1 of 2 diets groups (12 calves per group): (1) untreated basal diet (Control diet, CON), and (2) supplementation with 5% of FCGM in the basal diet (FCGM diet, FCGM, DM basis). Diets were isoenergetic and isonitrogenous as well as the amino acids levels were similar between 2 diets. Moreover, the calves received diets and water ad libitum throughout the 8 week trial period. Calves fed FCGM diet significantly increased the average daily gain (P = 0.028) and feed efficiency (P = 0.034) and tended to elevate withers height (P = 0.067) compared with the CON. No difference was observed for body weight, body length and heart girth between the treatments. The rumen ammonia nitrogen (P < 0.001), acetate (P = 0.004), propionate (P = 0.042), total volatile fatty acids (P = 0.002), microbial protein (P = 0.003) and proteinase (P = 0.001) were higher in FCGM calves. Moreover, rumen pH (P < 0.001) was lower in FCGM. The relative abundance of the phylum-level Bacteroidetes (79.4%) and the family-level Prevotellaceae (56.1%) in FCGM was higher than that in CON (66.8% and 37.7%, respectively). Feeding FCGM significantly increased the relative abundance of the genera-level Prevotella_1 (P = 0.028) and Prevotellaceae_UCG-003 (P = 0.016) but decreased the relative abundance of the Ruminobacter (P = 0.011) compared with the CON. Additionally, FCGM significantly increased the plasma total protein (P = 0.027), glucose (P = 0.014), total superoxide dismutase (P = 0.003), catalase (P = 0.023), total antioxidant capacity (P = 0.029), immunoglobulin A (P = 0.016), immunoglobulin G (P = 0.026), growth hormone (P = 0.027) and insulin-like growth factor-І (P = 0.007). In conclusion, the dietary inclusion of FCGM elevated the growth rate and feed efficiency, promoted rumen fermentation, diversified the bacterial community composition in the rumen, as well as improved antioxidant and immune functions of calves during the postweaning stage.

Expression Differences of Stress and Immunity Genes in Rainbow Trout (Oncorhynchus mykiss, Walbaum 1792) with Different Bacterial Fish Diseases

The aim of this study was to determine the changes in the mRNA transcription levels of HSP70 and IGF genes related to stress and immunity in Oncorhynchus mykiss obtained from fish farms, to determine the phenotypic and antimicrobial properties of the bacteria isolated in the study and to compare the results according to different diseases. Accordingly, six fish from each fish farm, a total of 30 fish were sampled. Bacterial identification, inoculations were carried out from the tissue samples taken from the kidney and symptomatic surfaces of fish samples in TSA. Primary cultures were obtained after incubation periods at 21 and 37oC. Gram staining, catalase, and oxidase tests were performed. Forty-six biochemical and 26 antimicrobial tests were performed using BD Phoenix ID Panels. Muscle tissue was used to determine mRNA gene expression differences and the tissues were preserved at -80°C in RNAlater storage solution. Following the RNA isolation and cDNA synthesis, a real-time PCR procedure was performed with b-Actin (ACTB), Insuline-like growth factor (IGF) and HSP70 Gene Primer Array system. Enterococcus faecium (E. faecium), Lactococcus garvieae (L. garvieae) and Staphylococcus aureus (S. aureus) agents were isolated in the study. These bacteria were identified with 91-99% similarity ratios. No disease agents in the gene expressions of the fish samples in were isolated and they were therefore used as controls. Compared to the control group, heat shock protein (HSP) mRNA expression was upregulated in fish tissues infected with E. faecium, S. aureus, and L. garvieae whereas no significant increase was determined in fish tissues infected with S. aureus. IGF mRNA gene expression was upregulated in all infected tissues. IGF expression was upregulated at the highest level in fish tissues infected with L. garvieae.

HSP70 Inhibitor Suppresses IGF-I-Stimulated Migration of Osteoblasts through p44/p42 MAP Kinase.

Heat shock protein 70 (HSP70) is a ubiquitously expressed molecular chaperone in a variety of cells including osteoblasts. We previously showed that insulin-like growth factor-I (IGF-I) elicits migration of osteoblast-like MC3T3-E1 cells through the activation of phosphatidylinositol 3-kinase/Akt and p44/p42 mitogen-activated protein (MAP) kinase. In the present study, we investigated the effects of HSP70 inhibitors on the IGF-I-elicited migration of these cells and the mechanism involved. The IGF-I-stimulated osteoblast migration evaluated by a wound-healing assay and by a transwell cell migration was significantly reduced by VER-155008 and YM-08, which are both HSP70 inhibitors. VER-155008 markedly suppressed the IGF-I-induced phosphorylation of p44/p42 MAP kinase without affecting that of Akt. In conclusion, our results strongly suggest that the HSP70 inhibitor reduces the IGF-I-elicited migration of osteoblasts via the p44/p42 MAP kinase.

A Polymorphism of Insulin-Like Growth Factor Binding Protein 2 Gene Associated with Growth and Body Composition Traits in Kampong Chickens

Insuline-like growth factor binding protein 2 (IGFBP2) is one of the principal binding proteins that has biological functions involved in growth, development, and differentiation. Selection for rapid growth based on molecular Marker Assisted Selection is required to increase production performance. The present study was designed to analyze the associations of IGFBP2 gene polymorphisms with chicken growth and body composition traits. Kampong chicken, a native chicken in Indonesia, is slow-growing chicken. A total of 59 males were used in the current study. Growth and body composition were measured in 24 wk of age. Primers for intron 2 region were designed from genomic chicken sequence. A c.1032C&gt;T SNP of the IGFBP2 gene intron 2 region was detected and PCR-RFLP method was then used to genotype Kampong chicken population. The result showed that IGFBP2 polymorphism was significantly associated with body, carcass, breast, breast muscle, pectoralis minor, leg, and wings weight in Kampong chicken population (P &lt; 0.05). The research suggests that the IGFBP2 gene could be a candidate gene that affects growth and body composition traits in chicken.

Influence of puberty timing on adiposity and cardiometabolic traits: A Mendelian randomisation study.

BackgroundEarlier puberty is widely linked with future obesity and cardiometabolic disease. We examined whether age at puberty onset likely influences adiposity and cardiometabolic traits independent of childhood adiposity.Methods and findingsOne-sample Mendelian randomisation (MR) analyses were conducted on up to 3,611 white-European female and male offspring from the Avon Longitudinal Study of Parents and Children (ALSPAC) cohort recruited at birth via mothers between 1 April 1991 and 31 December 1992. Time-sensitive exposures were age at menarche and age at voice breaking. Outcomes measured at age 18 y were body mass index (BMI), dual-energy X-ray absorptiometry–based fat and lean mass indices, blood pressure, and 230 cardiometabolic traits derived from targeted metabolomics (150 concentrations plus 80 ratios from nuclear magnetic resonance [NMR] spectroscopy covering lipoprotein subclasses of cholesterol and triglycerides, amino acids, inflammatory glycoproteins, and others). Adjustment was made for pre-pubertal BMI measured at age 8 y. For negative control MR analyses, BMI and cardiometabolic trait measures taken at age 8 y (before puberty, and which therefore cannot be an outcome of puberty itself) were used. For replication analyses, 2-sample MR was conducted using summary genome-wide association study data on up to 322,154 adults for post-pubertal BMI, 24,925 adults for post-pubertal NMR cardiometabolic traits, and 13,848 children for pre-pubertal obesity (negative control). Like observational estimates, 1-sample MR estimates in ALSPAC using 351 polymorphisms for age at menarche (explaining 10.6% of variance) among 2,053 females suggested that later age at menarche (per year) was associated with −1.38 kg/m2 of BMI at age 18 y (or −0.34 SD units, 95% CI −0.46, −0.23; P = 9.77 × 10−09). This coefficient attenuated 10-fold upon adjustment for BMI at age 8 y, to −0.12 kg/m2 (or −0.03 SDs, 95% CI −0.13, 0.07; P = 0.55). Associations with blood pressure were similar, but associations across other traits were small and inconsistent. In negative control MR analyses, later age at menarche was associated with −0.77 kg/m2 of pre-pubertal BMI measured at age 8 y (or −0.39 SDs, 95% CI −0.50, −0.29; P = 6.28 × 10−13), indicating that variants influencing menarche also influence BMI before menarche. Cardiometabolic trait associations were weaker and less consistent among males and both sexes combined. Higher BMI at age 8 y (per 1 kg/m2 using 95 polymorphisms for BMI explaining 3.4% of variance) was associated with earlier menarche among 2,648 females (by −0.26 y, 95% CI −0.37, −0.16; P = 1.16 × 10−06), likewise among males and both sexes combined. In 2-sample MR analyses using 234 polymorphisms and inverse variance weighted (IVW) regression, each year later age at menarche was associated with −0.81 kg/m2 of adult BMI (or −0.17 SD units, 95% CI −0.21, −0.12; P = 4.00 × 10−15). Associations were weaker with cardiometabolic traits. Using 202 polymorphisms, later menarche was associated with lower odds of childhood obesity (IVW-based odds ratio = 0.52 per year later, 95% CI 0.48, 0.57; P = 6.64 × 10−15). Study limitations include modest sample sizes for 1-sample MR, lack of inference to non-white-European populations, potential selection bias through modest completion rates of puberty questionnaires, and likely disproportionate measurement error of exposures by sex. The cardiometabolic traits examined were heavily lipid-focused and did not include hormone-related traits such as insulin and insulin-like growth factors.ConclusionsOur results suggest that puberty timing has a small influence on adiposity and cardiometabolic traits and that preventive interventions should instead focus on reducing childhood adiposity.

A cellular mechanism of muscle memory facilitates mitochondrial remodelling following resistance training.

Referring to the muscle memory theory, previously trained muscles acquire strength and volume much faster than naive muscles. Using extreme experimental models such as synergist ablation or steroid administration, previous studies have demonstrated that the number of nuclei increases when a muscle becomes enlarged, which serves as a cellular muscle memory mechanism for the muscle. In the present study, we found that, when rats were subjected to physiologically relevant resistance training, the number of myonuclei increased and was retained during a long-term detraining period. The acquired myonuclei were related to a greater degree of muscle hypertrophic and mitochondrial biogenesis processes following subsequent hypertrophic conditions. Our data suggest a cellular mechanism supporting the notion that exposing young muscles to resistance training would help to restore age-related muscle loss coupled with mitochondrial dysfunction in later life. Muscle hypertrophy induced by resistance training is accompanied by an increase in the number of myonuclei. The acquired myonuclei are viewed as a cellular component of muscle memory by which muscle enlargement is promoted during a re-training period. In the present study, we investigated the effect of exercise preconditioning on mitochondrial remodelling induced by resistance training. Sprague-Dawley rats were divided into four groups: untrained control, training, pre-training or re-training. The training groups were subjected to weight loaded-ladder climbing exercise training. Myonuclear numbers were significantly greater (up to 20%) in all trained muscles compared to untrained controls. Muscle mass was significantly higher in the re-training group compared to the training group (∼2-fold increase). Mitochondrial content, mitochondrial biogenesis gene expression levels and mitochondrial DNA copy numbers were significantly higher in re-trained muscles compared to the others. Oxidative myofibres (type I) were significantly increased only in the re-trained muscles. Furthermore, in vitro studies using insulin-like growth factor-1-treated L6 rat myotubes demonstrated that myotubes with a higher myonuclear number confer greater expression levels of both mitochondrial and nuclear genes encoding for constitutive and regulatory mitochondrial proteins, which also showed a greater mitochondrial respiratory function. These data suggest that myonuclei acquired from previous training facilitate mitochondrial biogenesis in response to subsequent retraining by (at least in part) enhancing cross-talk between mitochondria and myonuclei in the pre-conditioned myofibres.

Commensal Microbiota Enhance Both Osteoclast and Osteoblast Activities.

Recent studies suggest that the commensal microbiota affects not only host energy metabolism and development of immunity but also bone remodeling by positive regulation of osteoclast activity. However, the mechanism of regulation of bone cells by the commensal microbiota has not been elucidated. In this study, 8-week-old specific pathogen-free (SPF) and germ-free (GF) mice were compared in terms of alveolar bones and primary osteoblasts isolated from calvarias. Micro-CT analysis showed that SPF mice had larger body size associated with lower bone mineral density and bone volume fraction in alveolar bones compared with GF mice. Greater numbers of osteoclasts in alveolar bone and higher serum levels of tartrate-resistant acid phosphatase 5b were observed in SPF mice. Tissue extracts from SPF alveolar bone showed higher levels of cathepsin K, indicating higher osteoclast activity. SPF alveolar extracts also showed elevated levels of γ-carboxylated glutamic acid–osteocalcin as a marker of mature osteoblasts compared with GF mice. Polymerase chain reaction (PCR) array analysis of RNA directly isolated from alveolar bone showed that in SPF mice, expression of mRNA of osteocalcin, which also acts as an inhibitor of bone mineralization, was strongly enhanced compared with GF mice. Cultured calvarial osteoblasts from SPF mice showed reduced mineralization but significantly enhanced expression of mRNAs of osteocalcin, alkaline phosphatase, insulin-like growth factor-I/II, and decreased ratio of osteoprotegerin/receptor activator of nuclear factor-kappa B ligand compared with GF mice. Furthermore, PCR array analyses of transcription factors in cultured calvarial osteoblasts showed strongly upregulated expression of Forkhead box g1. In contrast, Gata-binding protein 3 was strongly downregulated in SPF osteoblasts. These results suggest that the commensal microbiota prevents excessive mineralization possibly by stimulating osteocalcin expression in osteoblasts, and enhances both osteoblast and osteoclast activity by regulating specific transcription factors.

Determination the Concentration of Insulin-Like Growth Factor-I (IGF-I) in Saliva of Acromegalic Patients and Comparison It with the Levels of Serum IGF-I.

Acromegaly is ametabolic disorder characterized by an acquired progressive somatic disfigurement, mainly involving the face, extremities and many other organs, that are associated with systemic manifestations, caused by excessive secretion of growth hormone and a resultant persistent elevation of insulin-like growth factor-I concentrations. In more than 90% of cases originates from a monoclonal benign pituitary adenoma. Aim of this study to assess the level of insulin-like growth factor-I (IGF-I) in saliva of acromegalic patients, and to compare it with the basal levels of serum IGF-I. Sixty specimens of serum and saliva collected from two groups of subjects (forty acromegalic patients and twenty healthy persons). The specimens were centrifuged and stored at -20ºC then IRMA kits were used for estimating insulin like-growth factor-I. The results show that acromegalic patients had significantly higher salivary insulin like growth factor-I concentrations than healthy subjects (mean 21.26 vs. 20.48ng/mL; p=0.041), serum insulin like growth factor-I concentrations (mean 782.21 vs. 199.87ng/mL; p

1128 Role of nucleotide excision repair pathway in insulin-like growth factor-1-mediated keratinocyte photoresponses

1128 Role of nucleotide excision repair pathway in insulin-like growth factor-1-mediated keratinocyte photoresponses

Clinical application and research progress in recombinant human growth hormone

Growth hormone is a protein secreted by the anterior pituitary acidophilic cell, which directly or indirectly affects growth and metabolism via insulin-like growth factor.Growth hormone can be applied to the target cells of various tissues, and has extensive physiological functions.With the development of further clinical studies, the application of recombinant human growth hormone is no longer limited to children dwarfism treatment, and it also has clinical significances in the adult growth hormone deficiency, neurological diseases such as hypoxic-ischemic encephalopathy, traumatic brain injuries, cerebral palsy, Alzheimer ′s disease and other diseases such as burn and assisted reproduction.This paper reviews current clinical application and research progress in recombinant human growth hormone at home and abroad. Key words: Recombinant human growth hormone; Adult growth hormone deficiency; Cerebral palsy

Abstract NTOC-097: VACCINATION TARGETING INSULIN–LIKE GROWTH FACTOR BINDING PROTEIN–2 (IGFBP–2) IN ADVANCED OVARIAN CANCER: SAFETY, IMMUNOGENICITY, AND SURVEILLANCE, EPIDEMIOLOGY, AND END RESULTS (SEER) COMPARISON

Abstract BACKGROUND: Immunization against self-antigens can induce regulatory responses that inhibit desirable Type 1 antitumor immune responses. Deletion of epitopes that favor a regulatory phenotype may improve the efficacy of therapeutic vaccines. We have developed a novel IGFBP-2 targeting DNA plasmid vaccine that selectively induces Type 1 immunity. IGFBP-2 regulates invasiveness and metastases in ovarian cancer. Eradication of ovarian cancer cells expressing IGFBP-2 through effective immunization could prevent disease relapse or metastasis. METHODS: Twenty-five patients with advanced stage or recurrent ovarian cancer treated to complete remission after primary or salvage therapy received 3 monthly doses of an IGFBP-2 DNA vaccine in a single-arm, non-randomized study. ELISPOT and flow cytometry were used to characterize antigen specific T-cell responses. Serum antibodies were measured using ELISA and Western blot. The SEER database was reviewed to identify women diagnosed between 2006 and 2012 matched for age, year of diagnosis and stage of diagnosis. The difference between dates of diagnosis and enrollment (lead time) was calculated for each patient receiving vaccine. Only SEER patients who survived at least as long as the lead time of their matches plus an additional 6 months were kept for analysis. In cases where this resulted in no SEER matched patients, unmatched vaccine patients were excluded. Overall survival (OS) was analyzed using Cox models and the Kaplan-Meier method. RESULTS: 206 adverse events (AE) were recorded. Fatigue (12%) and injection site reactions (12%) were the most common. 97% of AE were grades 1-2, 3% grade 3, and no grades 4 or 5. In preliminary immune analysis (16 patients), IGFBP-2 specific T-cell precursor frequencies are significantly elevated over baseline levels at 4 (p&amp;lt;0.01) and 6 (p&amp;lt;0.001) months. T-regulatory cells were not increased over the levels measured in a control reference population. No patients developed new IGFBP-2 specific antibody responses after immunization suggesting a lack of Th2 augmentation. Median OS for the matched SEER group (n=754) was 11 months. Matched IGFBP-2 vaccinated patients (n=20) have yet to reach median OS, but the lower 95% confidence limit is 27.3 months (p&amp;lt;0.0001). CONCLUSIONS: IGFBP-2 Th1 selective immunization is well tolerated, generates significant Type I immunity, and may demonstrate clinical efficacy. Citation Format: John B. Liao, Denise L. Cecil, Yushe Dang, Kelsey K. Baker, Kelsie J. Ovenell, Jessica Reichow, Stephanie Parker, Doreen M. Higgins, Jennifer S. Childs, Elizabeth K. Broussard, Andrew L. Coveler, Lupe G. Salazar, Barbara A. Goff, Mary W. Redman, Mary L. Disis. VACCINATION TARGETING INSULIN–LIKE GROWTH FACTOR BINDING PROTEIN–2 (IGFBP–2) IN ADVANCED OVARIAN CANCER: SAFETY, IMMUNOGENICITY, AND SURVEILLANCE, EPIDEMIOLOGY, AND END RESULTS (SEER) COMPARISON [abstract]. In: Proceedings of the 11th Biennial Ovarian Cancer Research Symposium; Sep 12-13, 2016; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2017;23(11 Suppl):Abstract nr NTOC-097.

Blood cadmium levels increase prostate specific antigen and insulin-like growth factor-1 among cadmium exposed workers

&lt;p&gt;BACKGROUND&lt;br /&gt;Cadmium (Cd) is a heavy metal that is classified as a human carcinogen (group IA), one of the cancers that it can cause being prostate cancer. The development of prostate cancer on a molecular basis involves oncogenes such as insuline-like growth factor-1 (IGF-1). Prostate cancer can be detected in the laboratory through the examination of prostate specific antigen (PSA). The present study aimed to determine the relationship of Cd levels with levels of PSA and IGF-1 in exposed and unexposed workers.&lt;/p&gt;&lt;p&gt;METHODS&lt;br /&gt;The study design was cross sectional. The subjects of the studycame from two groups of workers, ie. the group of Cd exposed workers who were welding shop workers and the group of unexposed workers who were office workers. The minimum samplesize was 85 people. The independent variable was blood Cd level. The dependent variables were PSA and IGF-1 levels. Blood Cd levels were measured by atomic absorption spectrometry (AAS), while PSA and IGF-1 were measured using ELISA. Data analysis was performed using the Mann-Whitney test and the Spearman correlation test.&lt;/p&gt;&lt;p&gt;RESULTS&lt;br /&gt;Mean blood Cd level in the exposed workers was 6.5 mg/L and in the unexposed workers 2.15 mg/L. There was a relationship between blood Cd and PSA levels (p&amp;lt;0.05) and between blood Cd levels and IGF-1 (p &amp;lt;0.05).&lt;/p&gt;&lt;p&gt;CONCLUSIONS&lt;br /&gt;There was a relationship of blood Cd with PSA and IGF-1 levels.among workers. PSA and IGF-1 could be a biochemical markers of disease control in cadmium exposed workers.&lt;/p&gt;

154 Lung cancer: Insulin-like growth factors/receptors

154 Lung cancer: Insulin-like growth factors/receptors

MiR-1275: A single microRNA that targets the three IGF2-mRNA-binding proteins hindering tumor growth in hepatocellular carcinoma

This study aimed to identify a single miRNA or miR (microRNA) which regulates the three insulin-like growth factor-2-mRNA-binding proteins (IGF2BP1, 2 and 3). Bioinformatics predicted miR-1275 to simultaneously target the three IGF2BPs, and screening revealed miR-1275 to be underexpressed in hepatocellular carcinoma (HCC) tissues. Transfection of HuH-7 cells with miR-1275 suppressed IGF2BPs expression and all three IGF2BPs were confirmed as targets of miR-1275. Ectopic expression of miR-1275 and knockdown of IGF2BPs inhibited malignant cell behaviors, and also reduced IGF1R protein and mRNA. Finally IGF1R was validated as a direct target of miR-1275. These findings indicate that the tumor-suppressor miR-1275 can control HCC tumor growth partially through simultaneously regulating the oncogenic IGF2BPs and IGF1R.

NFIL3 Suppresses Hypoxia‐induced Apoptotic Cell Death by Targeting the Insulin‐like Growth Factor 2 Receptor

The insulin-like growth factor-II/mannose 6-phosphate receptor (IGF2R) over-expression correlates with heart disease progression. The IGF2R is not only an IGF2 clearance receptor, but it also triggers signal transduction, resulting in cardiac hypertrophy, apoptosis and fibrosis. The present study investigated the nuclear factor IL-3 (NFIL3), a transcription factor of the basic leucine zipper superfamily, and its potential pro-survival effects in cardiomyocytes. NFIL3 might play a key role in heart development and act as a survival factor in the heart, but the regulatory mechanisms are still unclear. IGF2 and IGF2R protein expression were highly increased in rat hearts subjected to hemorrhagic shock. IGF2R protein expression was also up-regulated in H9c2 cells exposed to hypoxia. Over-expression of NFIL3 in H9c2 cardiomyoblast cells inhibited the induction of hypoxia-induced apoptosis and down-regulated IGF2R expression levels. Gel shift assay, double-stranded DNA pull-down assay and chromatin immune-precipitation analyses indicated that NFIL3 binds directly to the IGF2R promoter region. Using a luciferase assay, we further observed NFIL3 repress IGF2R gene promoter activity. Our results demonstrate that NFIL3 is an important negative transcription factor, which through binding to the promoter of IGF2R, suppresses the apoptosis induced by IGF2R signaling in H9c2 cardiomyoblast cells under hypoxic conditions.

P0269 : The insuline-like growth factor 2 (IGF2) MRNA binding protein (IMP) p62 promotes cirrhosis-linked hepatocarcinogenesis

P0269 : The insuline-like growth factor 2 (IGF2) MRNA binding protein (IMP) p62 promotes cirrhosis-linked hepatocarcinogenesis

The Role of the Active Oxygen Produced from Gp91phox NADPH Oxidase on the Newborn Weight of Mouse Pups

It is known that active oxygen plays an important role in a reproduction. However, no report has so far investigated the influence of active oxygen produced from gp91phox NADPH oxidase on newborns. In this study, we investigated the influence of active oxygen on the weight of newborns using graviditas gp91phox-knockout (gp91phox-/-) mice. Gestational C57BL/6j (control), gp91phox-/-, and insulin-like growth factor-1-knockout (IGF-1-/-) mice were examined and the weight of the newborn mouse pups were analyzed. Gp91phox-/- and IGF-1-/- mouse pups had low weight compared with control mice. When the control mice were treated with an inhibitor of reactive oxygen species (ROS), the newborn weight decreased. Conversely, when the gp91phox-/- mice were treated with an activator of ROS, the newborn weight increased, however, it remained low in the IGF-1-/- mice. Moreover, there were decreased levels of IL-1 in the plasma of graviditas gp91phox-/- mice compared with control and IGF-1-/- mice. Treatment with an IL-1 receptor antagonist in the control mice resulted in a low newborn weight, similar to the gp91phox-/- and IGF-1-/- mice. Furthermore, the expression of NLRP3 and caspase-1 in the uterus of graviditas gp91phox-/- mice was low compared with the control and IGF-1-/- mice. These results clearly indicate that gp91phox NADPH oxidase produces ROS during graviditas. The ROS activate NLRP3, and NLRP3 leads to the production of caspase-1, which subsequently increases IL-1, thereby finally inducing IGF-1. Because the newborn weight is determined by IGF-1, gp91phox appears to be important for promoting fetal growth during graviditas

Exploration of protective strategies against oligodendrocyte cell death in Krabbe disease models

Event Abstract Back to Event Exploration of protective strategies against oligodendrocyte cell death in Krabbe disease models Adrian Sandoval1, Jenny Jaramillo1, Diego Ordoñez1, David Martinez1 and Gonzalo Arboleda1* 1 Universidad Nacional de Colombia, Grupo de Muerte Celular, Departamento de Patología e Instituto de Genética, Colombia Krabbe disease (KD) patients accumulate psychosine (galactosylsphingosine), a cytotoxic metabolite for oligodendrocytes, inducing early demyelination. Apoptosis has been suggested that plays an important role in psychosine-induced oligodendrocytes cell death in culture and in brains of Krabbe patients and an animal model of the disease (twitcher mouse). However, the molecular mechanism that triggers the activation of the apoptotic pathway, and hence the development/progression of the disease, still is not well understood. Here we report that silencing GALC gene expression induces cell death of the human derived oligodendrocyte cell line MO3.13. The induction of cell death is associated with the activation of caspase 3 and increase in Bax expression, suggesting that mitochondria is compromise, and decrease in cell survival signaling pathways such as PI3K/AKT, MAPK/ERK and AMPK, as observed by western blot analysis, 2 days after silencing. The data suggests an important psychosine-induced deregulation in apoptotic and anti-apoptotic cellular pathways. Moreover, pre-treatment with insuline-like growth factor (IGF-1) and PPARalfa agonist (WY 14643), significantly provides protection against the psychosine-induced changes described. Our data indicates that oligodendrocytes have a marked susceptibility to endogenous accumulation of psychosine and identified potential compounds that may offer protection against psychosine-induced apoptosis in vivo. Acknowledgements Funded by Colciencias and DIB-Universidad Nacional de Colombia.

Wogonin suppresses melanoma cell B16-F10 invasion and migration by inhibiting Ras-medicated pathways.

The patients diagnosed with melanoma have a bad prognosis for early regional invasion and distant metastases. Wogonin (5,7-dihydroxy-8-methoxyflavone) is one of the active components of flavonoids that extracts from Scutellariae radix. Several previous studies reported that wogonin possesses antitumor effect against leukemia, gastrointestinal cancer and breast cancer. In this study, we used melanoma cell B16-F10 to further investigate the anti-invasive and anti-migratory activity of wogonin. Our date showed that wogonin caused suppression of cell migration, adhesion, invasion and actin remodeling by inhibiting the expression of matrix metalloproteinase-2 and Rac1 in vitro. Wogonin also reduced the number of the tumor nodules on the whole surface of the lung in vivo. Furthermore, the examination of mechanism revealed that wogonin inhibited Extracellular Regulated protein Kinases and Protein Kinase B pathways, which are both medicated by Ras. Insulin-like growth factor-1-induced or tumor necrosis factor-α-induced invasion was also inhibited by wogonin. Therefore, the inhibitory mechanism of melanoma cell invasion by wogonin might be elucidated.

TNFα inhibits IGFBP-3 through activation of p38α and casein kinase 2 in human retinal endothelial cells.

We recently reported a reciprocal relationship between tumor necrosis factor alpha (TNFα) and insulin-like receptor growth factor binding protein 3 (IGFBP-3) in whole retina of normal and IGFBP-3 knockout mice. A similar relationship was also observed in cultured retinal endothelial cells (REC). We found that TNFα significantly reduced IGFBP-3 levels and vice-versa, IGFBP-3 can lower TNFα and TNFα receptor expression. Since IGFBP-3 is protective to the diabetic retina and TNFα is causative in the development of diabetic retinopathy, we wanted to better understand the cellular mechanisms by which TNFα can reduce IGFBP-3 levels. For these studies, primary human retinal endothelial cells (REC) were used since these cells undergo TNFα-mediated apoptosis under conditions of high glucose conditions and contribute to diabetic retinopathy. We first cultured REC in normal or high glucose, treated with exogenous TNFα, then measured changes in potential signaling pathways, with a focus on P38 mitogen-activated protein kinase alpha (P38α) and casein kinase 2 (CK2) as these pathways have been linked to both TNFα and IGFBP-3. We found that TNFα significantly increased phosphorylation of P38α and CK2. Furthermore, specific inhibitors of P38α or CK2 blocked TNFα inhibition of IGFBP-3 expression, demonstrating that TNFα reduces IGFBP-3 through activation of P38α and CK2. Since TNFα and IGFBP-3 are key mediators of retinal damage and protection respectively in diabetic retinopathy, increased understanding of the relationship between these two proteins will offer new therapeutic options for treatment.