Study Protocol – Diabetes and related conditions in urban Indigenous people in the Darwin, Australia region: aims, methods and participation in the DRUID Study-4

<p><b>Copyright information:</b></p><p>Taken from "Study Protocol – Diabetes and related conditions in urban Indigenous people in the Darwin, Australia region: aims, methods and participation in the DRUID Study"</p><p>BMC Public Health 2006;6():8-8.</p><p>Published online 17 Jan 2006</p><p>PMCID:PMC1373687.</p><p>Copyright © 2006 Cunningham et al; licensee BioMed Central Ltd.</p>lients from the Northern Territory Department of Health and Community Services' (NTDHCS) Client-Master Index (CMI). DRUID and NTDHCS figures exclude people with missing or ineligible postal codes

Study Protocol – Diabetes and related conditions in urban Indigenous people in the Darwin, Australia region: aims, methods and participation in the DRUID Study-5

<p><b>Copyright information:</b></p><p>Taken from "Study Protocol – Diabetes and related conditions in urban Indigenous people in the Darwin, Australia region: aims, methods and participation in the DRUID Study"</p><p>BMC Public Health 2006;6():8-8.</p><p>Published online 17 Jan 2006</p><p>PMCID:PMC1373687.</p><p>Copyright © 2006 Cunningham et al; licensee BioMed Central Ltd.</p

Study Protocol – Diabetes and related conditions in urban Indigenous people in the Darwin, Australia region: aims, methods and participation in the DRUID Study-3

<p><b>Copyright information:</b></p><p>Taken from "Study Protocol – Diabetes and related conditions in urban Indigenous people in the Darwin, Australia region: aims, methods and participation in the DRUID Study"</p><p>BMC Public Health 2006;6():8-8.</p><p>Published online 17 Jan 2006</p><p>PMCID:PMC1373687.</p><p>Copyright © 2006 Cunningham et al; licensee BioMed Central Ltd.</p

Study Protocol – Diabetes and related conditions in urban Indigenous people in the Darwin, Australia region: aims, methods and participation in the DRUID Study-1

<p><b>Copyright information:</b></p><p>Taken from "Study Protocol – Diabetes and related conditions in urban Indigenous people in the Darwin, Australia region: aims, methods and participation in the DRUID Study"</p><p>BMC Public Health 2006;6():8-8.</p><p>Published online 17 Jan 2006</p><p>PMCID:PMC1373687.</p><p>Copyright © 2006 Cunningham et al; licensee BioMed Central Ltd.</p>ous clients from the Northern Territory Department of Health and Community Services' (NTDHCS) Client-Master Index (CMI)

Study Protocol – Diabetes and related conditions in urban Indigenous people in the Darwin, Australia region: aims, methods and participation in the DRUID Study-6

<p><b>Copyright information:</b></p><p>Taken from "Study Protocol – Diabetes and related conditions in urban Indigenous people in the Darwin, Australia region: aims, methods and participation in the DRUID Study"</p><p>BMC Public Health 2006;6():8-8.</p><p>Published online 17 Jan 2006</p><p>PMCID:PMC1373687.</p><p>Copyright © 2006 Cunningham et al; licensee BioMed Central Ltd.</p

Effect of serum from IR subjects (15 min) on Akt and GSK3β phosphorylation in differentiated human myotubes.

<p>Each bar is the mean of the results with sera from 7 IR subjects. Experiments were performed in triplicate for each individual. Ctr denotes controls. <i>Panel A</i>: Basal ⁴⁷³Ser Akt phosphorylation in the presence of 100 nM insulin or of different serum concentrations (5%, 10% and 20%) from IR subjects. Blots are representative western blots of ⁹Ser Akt phosphorylation and total Akt amounts. Data (mean ± s.d.) are expressed as fold change versus control condition (no insulin nor serum) set at 1 (^*<i>P</i><0.05 vs. control). <i>Panel B</i>: Basal ⁹Ser GSK3β phosphorylation in the presence of 100 nM insulin or of different serum concentrations (5%, 10% and 20%) from IR subjects. The blot is a representative western blots of ⁹Ser GSK3β phosphorylation. Data (mean ± s.d.) are expressed as fold change versus control condition (no insulin nor serum) set at 1 (^*<i>P</i><0.05 vs. control).</p

Interaction between IRS1−PI3K−Akt signaling pathway and mTOR.

<p>Upon insulin binding insulin receptor activates, through insulin receptor substrate 1 (IRS1), phosphatidylinositol-3-kinase (PI3K), which results in Akt phosphorylation at ³⁰⁸Thr residue via Phosphatidylinositol 4,5-bisphosphate (PIP2) to Phosphatidylinositol 3,4,5-bisphosphate (PIP3) conversion followed by Akt and PDK1 recruitment to plasma membrane (not shown). A factor J present in duodenal-jejunal conditioned medium activates, possibly via the tuberous sclerosis complex TSC1-TSC2, the mammlian target of rapamycin complex 2 (mTORC2). mTORC2 also appears to be regulated by the PI3K pathway and phosphorylates Akt at ⁴⁷³Ser residue. Through TSC1-TSC2 and the GTPase Rheb, Akt activates mTORC1 and its direct substrate S6K1. Akt also inhibits GSK3. S6K1, mTORC1 and GSK3 phosphorylate serine residues on IRS1, thus attenuating insulin signalling. Hypothetical signalling is denoted by dotted lines.</p

Effect of conditioned medium (CM) on p70 S6kinase and GSK3β phosphorylation in differentiated L6 myotubes.

<p>Each bar is the mean of experiments in triplicate with 3 to 5 different preparations of CM. Data are normalized by β-actin amount and expressed as fold change versus control condition (no insulin nor CM) set at 10. Data are mean ± s.d. <i>Panel A</i>: ³⁸⁹Thr p70 S6K1 phosphorylation. L6 myotubes were treated with Rapamycin or PP242 for 1 hour before 15 min incubation with 25 µg/ml of <i>db/db</i> CM. <i>db/db</i> CM proteins inhibited the basal ³⁸⁹Thr p70 S6K1 phosphorylation (⁺<i>P</i><0.05 vs. control no inhibitors). PP242 further reduced the phosphorylation level in both control (^*<i>P</i><0.01, Rapamycin and PP242 vs. no inhibitor) and <i>db/db</i> CM (^#<i>P</i><0.05 vs. no inhibitor). <i>Panel B</i>: ⁹Ser GSK3β phosphorylation after 15 min co-incubation with 25 µg/ml CM with or without insulin. <i>db/db</i> CM proteins inhibited the insulin-stimulated increase of ⁹Ser GSK3β phosphorylation (^*<i>P</i><0.05 vs. insulin-induced phosphorylation in control set at 100). Lower molecular weight bands in the blot suggest proteolysis of phosphorylated form of GSK3β upon <i>db/db</i> CM treatment. <i>Panel C</i>: Time course of the effect of 25 µg/ml <i>db/db</i> CM on ⁹Ser GSK3β phosphorylation and proteolysis. The <i>db/db</i>-derived conditioned medium did not significantly affect basal phosphorylation of GSK3β on ⁹Ser, while it induced a rapid degradation of GSK3β, measurable since 5 min of incubation. After 15 min of pretreatment with <i>db/db</i> CM, a washout for 15 min with medium replacement was able to fully correct the alterations induced by the <i>db/db</i> CM. <i>Panel D</i>: Pretreatment of L6 myotubes with Rapamycin or PP242 for 1 hour before incubation with 25 µg/ml CM from <i>db/db</i> for 15 min in the absence or presence of 100 nM insulin. mTOR inhibitors had no effect on proteolysis of phosphorylated GSK3β induced by <i>db/db</i> CM proteins either in the presence or absence of insulin.</p

Estimates of parameters (mean ± s.d. over the population) of Eqs. (1)-(2) used to fit IPITT data.

#<p><i>P</i><0.001, <i>db/db</i> 150 µg vs. control and Swiss 15 µg;</p>+<p><i>P</i><0.005, <i>db/db</i> 15 or 150 µg vs. Swiss 15 or 150 µg;</p><p>?<i>P</i><0.005, <i>db/db</i> 15 µg vs. control;</p>×<p><i>P</i><0.01, <i>db/db</i> 15 µg vs. Swiss 15 and 150 µg;</p>*<p><i>P</i><0.05, <i>db/db</i> 15 vs. <i>db/db</i> 150 µg;</p><p>°<i>P</i><0.05, <i>db/db</i> 150 µg vs. Swiss 150 µg;</p>&<p><i>P</i><0.05, <i>db/db</i> 150 µg vs. control and Swiss 15 µg.</p

Estimates of parameters (estimate ± s.d. of the estimate) of Eq. (3) used to fit the rate of deoxyglucose uptake in L6 cells.

*<p><i>P</i><0.00005, <i>db/db</i> vs. control;</p>+<p><i>P</i><0.02, <i>db/db</i> vs. control;</p>&<p><i>P</i><0.00005, <i>db/db</i> vs. Swiss;</p>#<p><i>P</i><0.01, <i>db/db</i> vs. Swiss.</p