Circulating insulin-like growth factors may contribute substantially to insulin receptor isoform A and insulin receptor isoform B signalling

Background

Only a fraction of circulating insulin-like activity is due to insulin itself. The aim of this study was to determine total serum insulin-like activity mediated via the insulin receptor isoform A (IR-A) and isoform B (IR-B) by using kinase receptor activation (KIRA) assays specific for the IR-A and IR-B.

Methods

The IR-A and IR-B KIRA assays use human embryonic kidney cells which have been transfected with the human IR-A or IR-B gene and quantify serum-mediated phosphorylation of the IR.

Results

Both IR KIRA assays were sensitive (detection limit 32 pmol/L) and precise (intra- and inter assay CV: <12% and <15%). The EC₅₀s of insulin, IGF-I and IGF-II were 1.4, 11.2 and 6.7 nmol/L for the IR-A KIRA assay, and 1.3, 31.0 and 15.7 nmol/L for the IR-B KIRA assay.The operational range of both assays allowed for determination of total insulin-like activity in human serum. Analysis of serum samples showed that there was a significant positive correlation between serum insulin-like and immunoreactive insulin concentrations (IR-A: r = 0.56, p = 0.01, IR-B: r = 0.68, p = 0.001). Importantly, addition of IGF-I or IGF-II antibodies to human serum samples could substantially decrease the endpoint signal in both KIRA assays.

Conclusions

We showed that serum IGF-I and IGF-II may substantially contribute to IR signalling. Since IR isoform specific KIRA assays also take into account the contribution of IGFs present in serum on IR signalling, they may help to gain more insight into the roles of IGF mediated IR-A and IR-B activation in health and disease.     

Silicone breast implants may contribute to early-onset fetal growth restriction

Clinical Rheumatology - There are many studies showing that silicone breast implants may affect lactation, but few analyzed whether these implants affect placentation. We observed that many mothers...     

Altered insulin-like growth factor-1 and nitric oxide sensitivities in hypertension contribute to vascular hyperplasia

Vascular medial thickening, a hallmark of hypertension, is associated with vascular smooth muscle cell (VSMC) hypertrophy and hyperplasia. Although the precise mechanisms responsible are elusive, we have shown that strain induced regulation of autocrine insulin-like growth factor-1 (IGF-1) and nitric oxide (NO) reciprocally modulate VSMC proliferation. Therefore, we investigated potential IGF-1 and NO abnormalities in young (10-week-old) spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY) and their respective VSMC ex vivo. The SHR had increased mean arterial pressure (173 ± 2 v 128 ± 3 mm Hg, n = 24, P < .05) but similar pulse pressures (31 ± 2 v 30 ± 3 mm Hg; P > .05) v WKY. The SHR exhibited increased aortic wall thickness in comparison with WKY (523 ± 16 v 355 ± 17μm; P < .05). No differences were seen in plasma combined NO₂ and NO₃ (NO_x) (0.48 ± 0.11 mmol/L for WKY v 0.58 ± 0.18 mmol/L for SHR) or plasma IGF-1 (1007 ± 28 ng/mL for WKY v 953 ± 26 ng/mL for SHR). Aortic VSMC from SHR displayed enhanced proliferation in comparison with WKY (P < .05). Underlying this enhanced proliferation was altered SHR VSMC sensitivity to the antiproliferative NO donor 2,2"[Hydroxynitrosohydrazono] bis-ethanimine (DETA-NO) (ID₅₀: 270 ± 20 mmol/L for SHR; 150 ± 11 mmol/L for WKY; P < .05). Basal cyclic guanosine monophosphate (cGMP) secretion from SHR VSMC was 65-fold greater than that seen from WKY (P < .001). In response to DETA-NO, cGMP secretion from SHR VSMC increased modestly (1.5-fold; P < .01), whereas treatment of WKY VSMC resulted in a 26-fold (P < .001) increase in cGMP. The SHR VSMC did not respond to exogenous IGF-1, whereas WKY VSMC exhibited a dose dependent increase in proliferation with IGF-1 (10⁻¹⁰ to 10⁻⁷ mol/L). These data suggest that VSMC hyperplasia in early hypertension is not reflected by imbalances in plasma IGF-1 or NO. Rather, altered SHR VSMC sensitivity to NO is likely responsible in part for the observed hyperproliferation seen in early stages of hypertension.     

Buffering of crucial functions by paleologous duplicated genes may contribute cyclicality to angiosperm genome duplication

Genome duplication followed by massive gene loss has permanently shaped the genomes of many higher eukaryotes, particularly angiosperms. It has long been believed that a primary advantage of genome duplication is the opportunity for the evolution of genes with new functions by modification of duplicated genes. If so, then patterns of genetic diversity among strains within taxa might reveal footprints of selection that are consistent with this advantage. Contrary to classical predictions that duplicated genes may be relatively free to acquire unique functionality, we find among both Arabidopsis ecotypes and Oryza subspecies that SNPs encode less radical amino acid changes in genes for which there exists a duplicated copy at a "paleologous" locus than in "singleton" genes. Preferential retention of duplicated genes encoding long complex proteins and their unexpectedly slow divergence (perhaps because of homogenization) suggest that a primary advantage of retaining duplicated paleologs may be the buffering of crucial functions. Functional buffering and functional divergence may represent extremes in the spectrum of duplicated gene fates. Functional buffering may be especially important during "genomic turmoil" immediately after genome duplication but continues to act approximately 60 million years later, and its gradual deterioration may contribute cyclicality to genome duplication in some lineages.     

Effects of titanium particle size on osteoblast functions in vitro and in vivo

The formation of titanium (Ti)-wear particles during the lifetime of an implant is believed to be a major component of loosening due to debris-induced changes in bone cell function. Radiographic evidence indicates a loss of fixation at the implant-bone interface, and we believe that the accumulation of Ti particles may act on the bone-remodeling process and impact both long- and short-term implant-fixation strengths. To determine the effects of various sizes of the Ti particles on osteoblast function in vivo, we measured the loss of integration strength around Ti-pin implants inserted into a rat tibia in conjunction with Ti particles from one of four size-groups. Implant integration is mediated primarily by osteoblast adhesion/focal contact pattern, viability, proliferation and differentiation, and osteoclast recruitment at the implant site in vivo. This study demonstrates the significant attenuation of osteoblast function concurrent with increased expression of receptor activator of nuclear factor kappaB ligand (RANKL), a dominant signal for osteoclast recruitment, which is regulated differentially, depending on the size of the Ti particle. Zymography studies have also demonstrated increased activities of matrix metalloproteinases (MMP) 2 and 9 in cells exposed to larger Ti particles. In summary, all particles have adverse effects on osteoblast function, resulting in decreased bone formation and integration, but different mechanisms are elicited by particles of different sizes.     

Autocrine stimulation by insulin-like growth factor 1 and insulin-like growth factor 2 mediates chondrocyte survival in vitro

OBJECTIVE: To determine the role of autocrine stimulation by insulin-like growth factor 1 (IGF-1) and IGF-2 in mediating chondrocyte survival and to determine whether chondrocytes from older individuals are more susceptible to cell death when IGF action is blocked. METHODS: Survival was assessed in human and monkey chondrocytes cultured in suspension in alginate under serum-free conditions. The role of IGFs in mediating survival was determined by treating cultures with neutralizing antibodies to IGF-1 and IGF-2, an antibody that blocks the type 1 IGF receptor, and antisense oligonucleotides to inhibit IGF-1 production. Survival was measured in chondrocyte cultures from young and old adult monkeys in the presence and absence of the IGF receptor blocking antibody and ceramide to induce cell death. RESULTS: Cell survival of >90% was noted when chondrocytes were cultured for as long as 107 days in alginate in a supplemented serum-free medium. Compared with controls, survival was significantly reduced by treatment with neutralizing antibodies to IGF-1 (25% cell death), neutralizing antibodies to IGF-2 (18% cell death), antibody to the IGF receptor (45% cell death), and IGF-1 antisense oligonucleotides (28% cell death). Cell death from inhibition of the type 1 IGF receptor was associated with an increase in caspase 3 activity and with positive DNA fragmentation, consistent with apoptotic cell death. Chondrocytes from old adult monkeys were more susceptible to cell death than were those from young adult monkeys when the IGF receptor was blocked and cell death was further stimulated by ceramide. CONCLUSION: Autocrine production of IGFs helps to maintain chondrocyte survival in vitro and could play a similar role in vivo. With aging, chondrocytes may become more susceptible to factors that induce cell death.     

Vascular endothelial growth factor may contribute to increased vascular permeability in acute respiratory distress syndrome.

The development of noncardiogenic pulmonary edema is a characteristic feature of acute respiratory distress syndrome (ARDS). We hypothesized that vascular endothelial growth factor (VEGF) would play an important role in this process. Plasma VEGF was measured in 40 patients with ARDS, 28 at-risk patients, 14 normal control subjects, and 9 ventilated control subjects. Cultured peripheral blood mononuclear cells (PBM) supernatant VEGF was measured in 21 patients with ARDS and 12 at-risk patients, respectively. The functional importance of VEGF as a mediator of endothelial permeability was assessed by measuring albumin flux across human pulmonary endothelial cell monolayers. Plasma VEGF was significantly elevated in patients with ARDS compared with at-risk patients, normal control subjects, and ventilated control subjects (p = 0.01, p = 0.0001, and p = 0.002, respectively). PBM from patients with ARDS produced significantly more VEGF in vitro than at-risk patients (p = 0.05). Albumin flux across human pulmonary endothelial cell monolayers was significantly increased following the addition of plasma from patients with ARDS compared with plasma from normal control subjects (p = 0.008). When VEGF activity in plasma was neutralized by the addition of a soluble VEGF inhibitor, the albumin flux induced by ARDS plasma was reduced by 48%. We conclude that VEGF makes a significant contribution to the endothelial cell permeability-inducing activity in plasma from patients with ARDS, and may play an important role in the development of noncardiogenic pulmonary edema in ARDS.     

The insulin-like growth factor system and its pleiotropic functions in brain

In recent years, much interest has been devoted to defining the role of the IGF system in the nervous system. The ubiquitous IGFs, their cell membrane receptors, and their carrier binding proteins, the IGFBPs, are expressed early in the development of the nervous system and are therefore considered to play a key role in these processes. In vitro studies have demonstrated that the IGF system promotes differentiation and proliferation and sustains survival, preventing apoptosis of neuronal and brain derived cells. Furthermore, studies of transgenic mice overexpressing components of the IGF system or mice with disruptions of the same genes have clearly shown that the IGF system plays a key role in vivo.     

The role of insulin-like growth factor I in age-related changes in calcium homeostasis in men

The aim of this study was to evaluate hormonal influences on age-related changes in calcium homeostasis in men. We recruited 178 healthy men, ages 20-79 (about 30 per decade). We measured serum calcium, phosphate, urinary calcium, and creatinine clearance. Dietary calcium intake and use of fish oils were determined by questionnaire. Fractional calcium absorption was estimated using the stable strontium technique in a subgroup of 60 men. PTH, 1, 25-dihydroxyvitamin D [1,25(OH)(2)D], 25-hydroxyvitamin D (25OHD), serum insulin-like growth factor I (IGF-I), and testosterone were measured in all men. There was no change in serum calcium with age. There were decreases in serum phosphate, urinary calcium, and creatinine clearance with age (P: < 0.02). Dietary calcium was unchanged. Strontium absorption decreased (P: < 0.01), and PTH increased (P: < 0.001) with age. The data for 1,25OH(2)D were biphasic, reaching a peak at age 55 yr (P: = 0.003). There was a linear increase in 25OHD with age (P: = 0.009) that persisted after correcting for seasonal variation and was positively associated with fish oil use, therefore, the age-related changes in 25OHD were masked by self medication. There were log-linear decreases in IGF-I and testosterone with age (P: < 0.0001). Strontium absorption was not related to 25OHD or 1,25(OH)(2)D, but was positively correlated with IGF-I. 1,25(OH)(2)D correlated negatively with serum phosphate and calcium, but not PTH or creatinine clearance. IGF-I was positively associated with creatinine clearance, serum calcium, and phosphate and negatively associated with PTH (P: < 0.001). In this cross-sectional study of otherwise healthy, normally aging men, age-related decreases in IGF-I seem to have a greater impact on mineral absorption than does vitamin D status.     

Selective pressure for a decreased rate of asymmetrical divisions within stem cell niches may contribute to age-related alterations in stem cell function

Most mammalian continuously renewing tissues are maintained by stem cells located within stem cell niches. Each niche contains a number of stem cells that replicate asymmetrically to give differentiated cells and also undergo periodic deletion and compensatory replacement by symmetrical "self-renewal" divisions of stem cells remaining within the niche. It has been recognized that there is selective pressure for an increased rate of self-renewal divisions and that the increasingly likely consequence is neoplasia. However, to date it has not been appreciated that there is also an independent selective pressure for a decreased rate of asymmetrical divisions. In this article, the origin of this second type of selective pressure is explained and its consequences explored through the use of computer modeling. It is shown that age-related changes in a range of mammalian stem cell compartments can be understood in the context of a decreased rate of asymmetrical stem cell divisions with an increased propensity for self-renewal divisions. It is proposed that a decreased rate of asymmetrical divisions impairs the ability of old stem cell compartments to respond effectively to stress.     

Antibiotic selection may contribute to increases in macrolide-resistant Treponema pallidum

To determine whether the 23S rRNA mutation that confers macrolide resistance is present in >1 Treponema pallidum strain, 58 isolates collected between 2001 and 2005 were screened for this mutation and for an unrelated sequence that distinguishes between strains. The odds of identifying a macrolide-resistant strain increased over time (P=.006). In subjects who had received macrolides in the previous year, the relative risk of harboring a resistant strain was 2.2 (95% confidence interval, 1.1-4.4; P=.02). The macrolide-resistant strains were not identical. These findings suggest that macrolide resistance may be increasing in multiple strains in response to antibiotic pressure.     

Activation of osteoblast insulin-like growth factor-II/cation-independent mannose-6-phosphate receptors by specific phosphorylated sugars and antibodies induce insulin-like growth factor-II effects.

The phosphorylated monosaccharide, mannose-6-phosphate (M6P), causes a dose-dependent stimulation of alkaline phosphatase production by osteoblasts. The concentrations tested ranged from 0.1 to 30 mM. A maximal effect was reproducibly seen at 10-30 mM, and represented a 30% stimulation over control cells. Glucose-6-phosphate and fructose-1-phosphate also stimulated osteoblast alkaline phosphatase production, but not to the same extent as M6P. Sugar residues such as mannose, mannose-1-phosphate, and fructose-6-phosphate had no effect. The stimulatory effect of M6P is similar to that seen with insulin-like growth factor II(IGF-II). However, increasing doses of IGF-II did not further stimulate or add to the effect of 10 mM M6P. These data indicate that the mechanism for the transduction of the stimulatory signal may be similar for both IGF-II and M6P. They do not address, however, the possibility of separate or similar binding sites for the two agents. A specific polyclonal antibody to the IGF-II/cation-independent mannose-6-phosphate receptor (IGF-II/CI-MPR) elicits the same effects as M6P and IGF-II in these bone cells. Non-immune serum used as a control does not have any effect. These results suggest that activation of the osteoblast IGF-II/CI-MPR by either M6P or a specific antibody can evoke a biological response similar to that observed with IGF-II.     

Reduced plasma adiponectin concentrations may contribute to impaired insulin activation of glycogen synthase in skeletal muscle of patients with type 2 diabetes

Aims/hypothesis Circulating levels of adiponectin are negatively associated with multiple indices of insulin resistance, and the concentration is reduced in humans with insulin resistance and type 2 diabetes. However, the mechanisms by which adiponectin improves insulin sensitivity remain unclear.Subjects and methods Combining euglycaemic–hyperinsulinaemic clamp studies with indirect calorimetry and skeletal muscle biopsies, we examined the relationship between plasma adiponectin and parameters of whole-body glucose and lipid metabolism, and muscle glycogen synthase (GS) activity in 51 Caucasians (ten lean, 21 obese and 20 with type 2 diabetes).Results Plasma adiponectin was significantly reduced in type 2 diabetic compared with obese and lean subjects. In lean and obese subjects, insulin significantly reduced plasma adiponectin, but this response was blunted in patients with type 2 diabetes. Plasma adiponectin was positively associated with insulin-stimulated glucose disposal (r=0.48), glucose oxidation (r=0.54), respiratory quotient (r=0.58) and non-oxidative glucose metabolism (r=0.38), and negatively associated with lipid oxidation during insulin stimulation (r=−0.60) after adjustment for body fat (all p<0.01). Most notably, we found a positive association between plasma adiponectin and insulin stimulation of GS activity in skeletal muscle (r=0.44, p<0.01).Conclusions/interpretation Our results indicate that plasma adiponectin may enhance insulin sensitivity by improving the capacity to switch from lipid to glucose oxidation and to store glucose as glycogen in response to insulin, and that low adiponectin may contribute to impaired insulin activation of GS in skeletal muscle of patients with type 2 diabetes.     

Longitudinal bone growth in vitro: effects of insulin-like growth factor I and growth hormone

Longitudinal growth was studied using an in vitro model system of intact rat long bones. Metatarsal bones from 18- and 19-day-old rat fetuses, entirely (18 days) or mainly (19 days) composed of chondrocytes, showed a steady rate of growth and radiolabelled thymidine incorporation for at least 7 days in serum-free media. Addition of recombinant human insulin-like growth factor-I to the culture media resulted in a direct stimulation of the longitudinal growth. Recombinant human growth hormone was also able to stimulate bone growth, although this was generally accomplished after a time lag of more than 2 days. A monoclonal antibody to IGF-I abolished both the IGF-I and GH-stimulated growth. However, the antibody had no effect on the growth of the bone explants in control, serum-free medium. Unlike the fetal long bones, bones from 2-day-old neonatal rats were arrested in their growth after 1-2 days in vitro. The neonatal bones responded to IGF-I and GH in a similar fashion as the fetal bones. Thus in this study in vitro evidence of a direct effect of GH on long bone growth via stimulating local production of IGF by the growth plate chondrocytes is presented. Furthermore, endogenous growth factors, others than IGFs, appear to play a crucial role in the regulation of fetal long bone growth.     

Vitamin D receptor polymorphisms may contribute to asthma risk

Background: Asthma is a common chronic airway disorder associated with significant morbidity and mortality. Objective: Current study aims at investigating the correlation between four vitamin D receptor (VDR) gene polymorphisms and asthma susceptibility by conducting a meta-analysis. Methods: PubMed, EBSCO, Ovid, Wiley, Web of Science, Wanfang, CNKI and VIP databases were searched using combinations of keywords relating to VDR and asthma. The published studies were filtered using our stringent inclusion and exclusion criteria, and the resultant high-quality data from final selected studies were analyzed using Stata 12.0 software. Results: A total of 77 studies were initially retrieved, and after further selection, 9 studies were eligible in current analysis. The selected studies contained 2,116 patients with asthma and 1,884 healthy controls. Our results demonstrated that rs2228570, rs7975232 and rs731236 in both allele models and dominant models, and rs3782905 in allele model in the VDR gene were linked with a high risk of asthma. No significant association between VDR gene rs3782905 in dominant model and risk of asthma was detected. Conclusions: This meta-analysis provides convincing evidence that rs2228570, rs7975232, rs731236 and rs3782905 gene polymorphisms in VDR are associated with increased susceptibility to asthma, indicating VDR polymorphisms could be developed as biomarkers for asthma susceptibility.     

Attenuated in vitro coronary arteriolar vasorelaxation to insulin-like growth factor I in experimental hypercholesterolemia.

Insulin and insulin-like growth factor (IGF) 1 affect coronary vasoactivity. Experimental hypercholesterolemia is associated with coronary atherogenesis and altered vasomotor regulation. Because the IGF axis is altered during atherogenesis, we postulated that experimental hypercholesterolemia is associated with an altered coronary vasoactive response to IGF-1 in vitro. Coronary arteries and arterioles from pigs fed either a normal or high-cholesterol diet for 10 weeks were contracted with endothelin-1 and relaxed with cumulative concentrations of insulin or IGF-1 (10(-12) to 10(-7) mol/L). Control arterioles were also incubated with the nitric oxide synthase inhibitor 10(-4) mol/L N(G)-monomethyl-L-arginine (L-NMMA) or the potassium channel blocker 10(-2) mol/L tetraethylammonium (TEA), contracted with endothelin-1, and relaxed with insulin or IGF-1. Experimental hypercholesterolemia (1) increased serum cholesterol (9.5+/-1.0 versus 1.9+/-0.08 mmol/L; P<0.0001), (2) caused coronary arterial and arteriolar endothelial dysfunction in vitro (attenuated vasorelaxation to bradykinin), (3) did not alter the epicardial response to either insulin (P=0.80) or IGF-1 (P=0.12), and (4) significantly attenuated the arteriolar response to IGF-1 (maximal relaxation of 79+/-6% versus 42+/-8%; P=0.01) but not insulin (43+/-6% versus 53+/-7%; P=0.99). Control arteriolar vasorelaxation to IGF-1 was attenuated by both L-NMMA (P<0.001) and TEA (P=0.01), whereas only L-NMMA attenuated insulin (P<0.001). Staining for IGF-1 and IGF binding protein 2 was increased (P<0.05) in arterioles of cholesterol-fed pigs. IGF-1 and insulin are therefore coronary arteriolar vasorelaxants through different mechanisms. Experimental hypercholesterolemia is associated with resistance to the coronary arteriolar vasorelaxing effects of IGF-1 but not insulin, in conjunction with increased ligand and binding-protein expression. The IGF axis may contribute to the altered coronary vasoactivity in hypercholesterolemia.     

Changes in the nitric oxide system contribute to effect of procyanidins extracted from the lotus seedpod ameliorating memory impairment in cognitively impaired aged rats

The major purpose of this study was to determine the effect of procyanidins extracted from the lotus seedpod (LSPC) on the nitric oxide (NO) system in the hippocampus and cerebral cortex in cognitively impaired aged rats. Using the Morris water maze, aged-unimpaired (AU) and aged-impaired (AI) rats were chosen from aged rats. All aged rats exhibited elevated inducible nitric oxide synthase (iNOS) activities and decreased neuronal nitric oxide synthase (nNOS) activities in the both brain regions. The changes were more pronounced in the brain of AI rats, especially in the hippocampus. Furthermore, AI rats greatly lowed the percentage of change of hippocampal nNOS activity in the presence of protein kinase inhibitors or phosphatase inhibitor, which meant that AI animals existed in a hardly modified nNOS dephosphorylated state in hippocampus. LSPC supplementation [50, 100 mg/kg of body weight (BW), per os (p.o.)] for 7 weeks significantly decreased iNOS activities and improved hippocampal nNOS phosphorylation status in AI animals. These results suggested that changes in the NO system may involve in the ameliorative effects of LSPC on cognitive deficits in AI animals.