Source-specific probabilistic health risk assessment of heavy metals in surface water of the Yangtze River Basin.

The detrimental effects of heavy metal accumulation on both ecosystems and public health have raised widespread concern. Source-specific risk assessment is crucial for developing effective strategies to prevent and control heavy metal contamination in surface water. This study aims to investigate the contamination characteristics of heavy metals in the Yangtze River Basin, identifying the pollution sources, assessing the risk levels, and further evaluating the health risks to humans. The results indicated that the average concentrations of heavy metals were ranked as follows: zinc (Zn) > arsenic (As) > copper (Cu) > chromium (Cr) > cadmium (Cd) > nickel (Ni) > lead (Pb), with average concentrations of 38.02 µg/L, 4.34 µg/L, 2.53 µg/L, 2.10 µg/L, 1.17 µg/L, 0.84 µg/L, and 0.32 µg/L, respectively, all below the WHO 2017 standards for safe drinking water. The distribution trend indicates higher concentrations in the upper and lower reaches and lower concentrations in the mid-reaches of the river. By integrating the Absolute Principal Component Scores-Multiple Linear Regression (APCS-MLR) receptor model and Positive Matrix Factorization (PMF) model, the main sources of heavy metals were identified as industrial activities (APCS-MLR: 41.3 %; PMF: 42.1 %), agricultural activities (APCS-MLR: 30.1 %; PMF: 27.4 %), and unknown mix sources (APCS-MLR: 29.1 %; PMF: 30.4 %). The calculation of the hazard index (HI) for both children and adults was <1, indicating no non-carcinogenic or carcinogenic risks. Based on the source-specific risk assessment, agricultural activities contribute the most to non-carcinogenic risks, while industrial activities pose the greatest contribution to carcinogenic risks. This study offers a reference for monitoring heavy metals and controlling health risks to residents, and provides crucial evidence for the utilization and protection of surface water in the Yangtze River Basin.

Tracing sulfate sources and transformations of surface water using multiple isotopes in a mining-rural-urban agglomeration area.

Rapid urbanization and mining activities are exacerbating sulfate (SO42-) pollution in surface water, and the information on its sources and transformations is crucial for understanding the sulphur cycle in mining areas. In this study, the SO42- in the surface water of Huaibei mining area were monitored and the main sources of pollution and biogeochemical processes were identified using stable isotopes (δD, δ18O-H2O, δ34S-SO42- and δ18O-SO42-) and water chemistry. The results demonstrated the SO42- content in the Huihe River and Linhuan subsidence water area (SWA) is higher than that in other rivers and SWAs, which exceeded the environmental quality standard of surface water. The SO42- content of different rivers and SWAs showed seasonal differences, and the dry season was higher than the wet season. In addition, the SO42- in Tuohe River and Suihe River is primarily caused by urban sewage and agriculture activities, while in Zhonghu and Shuoxihu SWA is mainly contributed by natural evaporate dissolution. Notably, the input of SO42- in the Huihe River and Linhuan SWA caused by mining activities cannot be disregarded. The aerobic environment and isotopic fractionation of surface water indicate that sulfide oxidation is not the major cause of SO42- formation. This work has revealed the multiple sources and transformation mechanisms of SO42-, and provided a reference for the development of comprehensive management and effective remediation strategies of SO42- contamination in surface water around mining areas.

Identification of nitrate accumulation mechanism of surface water in a mining-rural-urban agglomeration area based on multiple isotopic evidence.

The accumulation of nitrate (NO3-) in surface waters resulting from mining activities and rapid urbanization has raised widespread concerns. Therefore, it is crucial to develop a nitrate transformation information system to elucidate the nitrogen cycle and ensure sustainable water quality management. In this study, we focused on the main river and subsidence area of the Huaibei mining region to monitor the temporal and spatial variations in the NO3- content. Multiple isotopes (δD, δ18O-H2O, δ15N-NO3-, δ18O-NO3-, and δ15N-NH4+) along with water chemistry indicators were employed to identify the key mechanisms responsible for nitrate accumulation (e.g., nitrification and denitrification). The NO3- concentrations in surface water ranged from 0.28 to 7.50 mg/L, with NO3- being the predominant form of nitrogen pollution. Moreover, the average NO3- levels were higher during the dry season than during the wet season. Nitrification was identified as the primary process driving NO3- accumulation in rivers and subsidence areas, which was further supported by the linear relationship between δ15N-NO3- and δ15N-NH4+. The redox conditions and the relationship between δ15N-NO3- and δ18O-NO3-, and lower isotope enrichment factor of denitrification indicated that denitrification was weakened. Phytoplankton preferentially utilized available NH4+ sources while inhibiting NO3- assimilation because of their abundance. These findings provide direct evidence regarding the mechanism underlying nitrate accumulation in mining areas, while aiding in formulating improved measures for effectively managing water environments to prevent further deterioration.

Basin-wide tracking of nitrate cycling in Yangtze River through dual isotope and machine learning.

The nitrate (NO3-) input has adversely affected the water quality and ecological function in the whole basin of the Yangtze River. The protection of water sources and implementation of "great protection of Yangtze River" policy require large-scale information on water contamination. In this study, dual isotope and Bayesian mixing model were used to research the transformation and sources of nitrate. Chemical fertilizers contribute 76 % of the nitrate sources in the upstream, while chemical fertilizers were also dominant in the midstream (39 %) and downstream (39 %) of Yangtze River. In addition, nitrification process occurred in the whole basin. Four machine learning models were used to relate nitrate concentrations to explanatory variables describing influence factors to predict nitrate concentrations in the whole basin of Yangtze River. The anthropogenic and natural factors, such as rainfall, GDP and population were chosen to take as predictor variables. The eXtreme Gradient Boosting (XGBoost) model for nitrate has a better predictive performance with an R2 of 0.74. The predictive models of nitrate concentrations will help identify the nitrate distribution and transport in the whole Yangtze River basin. Overall, this study represents the first basin-wide data-driven assessment of the nitrate cycling in the Yangtze River basin.

Occurrence and source analysis of heavy metals and dissolved organic matter in Nanyi Lake, Anhui Province.

Nanyi Lake is a tributary in the lower reaches of the Yangtze River and the largest freshwater lake in southern Anhui. Anthropogenic activities have significantly affected the lake with the rapid development of the surrounding economy in recent years. This study collected a total of 39 × 2 water samples of surface water and overlying water, detected the heavy metal concentrations (As, Cr, Cu, Mn, Ni, Pb, Sb, Zn, Fe, Cd), and analyzed the spectral characteristics of the dissolved organic matter (DOM). The results demonstrated that the concentrations of heavy metals (As, Cr, Cu, Mn, Ni, Pb, Sb, Zn, Fe, Cd) in the surface water were 1.00-7.78, 0.40-5.59, 0.20-4.52, 20.00-269.50, 0.40-5.56, 0.20-5.06, 1.00-7.64, 20.00-252.50, 60.00-590.00, and 0.04-0.60, respectively (unit: µg L-1). The risk assessment showed that the carcinogenic risks of heavy metals (As, Cr, Cd) through drinking water were ranged from 10-7 to 10-4a-1 in Nanyi Lake, and Cr was the main carcinogen which should be the focus of environmental health risk management. The average personal non-carcinogenic risks of heavy metals (Cu, Mn, Ni, Pb, Sb, Zn, Fe) were ranged from 10-9 to 10-13a-1, and considered to be acceptable risk level. The contour map of spatial distribution demonstrated different degrees of heavy metals (except Zn) enrichment near the Langchuan River in the East Lake District. Parallel factor analysis showed that the main components of DOM in Nanyi Lake were tryptophan like, fulvic acid like, and tyrosine like, and the dissolved organic matter was primarily derived from autogenous endogenous sources. The heavy metals Cr, Ni, Pb, Fe, Cd, and HIX in the surface water of West Lake were significantly positively correlated, among which Ni, Fe and C2 were significantly negatively correlated (P < 0.05), Mn and BIX demonstrated extremely significant positive correlations (P < 0.01), while no significant correlation was observed between heavy metals and the DOM indexes in the surface water of the East Lake. The principal component and correlation analysis showed that the heavy metals in Nanyi Lake were primarily derived from the production wastewater discharged by the surrounding industrial and mining enterprises through the rivers input, followed by the non-point source runoff input of the surrounding agricultural production and lake aquaculture.

Distribution, Source Apportionment and Risk Assessment of Polycyclic Aromatic Hydrocarbons (PAHs) in Surface Sediments at the Basin Scale: A Case Study in Taihu Basin, China.

As a systematic research at basin scale, this study investigated the spatial distribution, source apportionment and ecological risks of eighteen polycyclic aromatic hydrocarbons (PAHs) in surface sediments at different functional regions (rivers, lakes and reservoirs) from Taihu basin. Results showed that the mean values of 18 PAHs (defined as ∑18PAHs) in river sediments (1277 ng/g) was much higher than those observed in lake sediments (243 ng/g) and reservoir sediments (134 ng/g). The accumulation of PAHs in river sediments was largely impacted by the local social-economic development and energy consumption. The positive matrix factorization (PMF) and isomer ratios analysis of PAHs suggest that relative contributions to PAHs in sediments were 15% for gasoline and heavy oil combustion, 9% for oil spills, 30% for coal combustion, 23% for traffic source, and 23% for diagenetic source. Ecological risk assessment based upon risk quotient (RQ) method indicated that sediments at Taihu basin have suffered moderate risk of PAHs.

Characterization of phosphorus forms in a Eutrophic Lake, China.

In this study, molecular compositions of cyanobacteria, suspended matter, and surface sediments in the Dianchi Lake, a highly eutrophic lake, were investigated by solution and solid-state 31P nuclear magnetic resonance (NMR) spectroscopy. A solution-state 31P NMR spectral analysis of NaOH-EDTA-extracted samples revealed the presence of orthophosphate (ortho-P, 48.5%-91.2% of the total extracted phosphate), orthophosphate monoester (mono-P, 7.3%-43.9%), orthophosphate diester (diester-P, 0.9%-3.9%), and pyrophosphate (pyro-P, 0.7%-5.5%). The organic phosphorus (Po) distribution in suspended matters and cyanobacteria was relatively similar but different from surface sediments. The inorganic phosphorus (Pi) distribution in suspended matters and surface sediments was slightly similar. Results of the solid-state 31P NMR spectral analysis of non-extractable residue showed that cyanobacteria and suspended matter contain a large proportion of Po and poly-P. For surface sediment, only metal-bonded pyro-P and a high proportion of ortho-P were detected. The solid-state 31P NMR spectra results of extracted residual indicated that several of the pyro-P bound to metals and other Pi or Po compounds associated with mineral phases in suspended matter and surface sediment are non-extractable. This result revealed that a high proportion of biogenic phosphorus is bioavailable. These results verify the process of biogenic poly-P regeneration in the Dianchi Lake given that Po, poly-P, and pyro-P are predominant in cyanobacteria and can be released into lake water through chemical and biological degradation, thus further worsening eutrophication.

Occurrence of phosphorus, iron, aluminum, silica, and calcium in a eutrophic lake during algae bloom sedimentation.

Phosphorus (P) in a water body is mainly controlled by the interaction between surface sediment and the overlying water column after the complete control of external pollution. Significant enhancement of P in a water body would cause eutrophication of lakes. Thus, a better understanding is needed of the occurrences of P between the sediment and water column in eutrophic lakes. Here, we measured total phosphorus (TP) and major elements (Fe, Al, Ca, Mn, Si) in the water column, and total nitrogen, organic matter, TP and major oxides (Fe2O3, Al2O3, CaO, SiO2) in surface sediment of Chaohu Lake, a continuously eutrophic lake. The results showed that the rank of TP levels was western lake > eastern lake > southern lake. There were significantly positive correlations between TP (including water TP and sedimentary TP) and Fe, Al, Mn, while the correlation coefficients between water TP and sedimentary TP were -0.43, -0.41 and 0.18 for the western, eastern and southern lake respectively. The negative and significant correlations of water TP and sedimentary TP may indicate that the risk of sedimentary P release was great in the western and eastern lake during algae bloom sedimentation, while the southern lake showed weak P exchange between the sediment and water column.

[The analysis of surgery effect of OSAHS children accompanied with sinusitis].

OBJECTIVE: To discuss the clinical efficacy of adenoidectomy and tonsillectomy for the OSAHS children with sinusitis. METHOD: Reviewing 112 cases of children with OSAHS, in which the 80 patients without chronic sinusitis, 32 ones with chronic sinusitis. Among them, 103 cases of children with adenoidectomy, tonsil resection, 9 cases adenoidectomy only. We did questionnaire survey before and after surgeryand also did post-surgery clinical follow-up. RESULT: The effective rate was 90.0% for the OSAHS children without sinusitis after adenoid and tonsil ectomy surgery, while the effective rate was 46.9% for the ones with sinusitis, the efficiency was significant different in the two groups after surgery; the 16.1% children after tonsil ectomy surgery complained the incidence of pharyngeal foreign body sensation and pharyngeal lymphoid hyperplasia in children was 23.2%. CONCLUSION: The efficiency was aviable for the OSAHS children without sinusitis after surgery; The efficiency was not aviable enough for the OSAHS children with sinusitis after surgery, so the latter one need adjuvant drug treatment; We also should be attention to the pharyngeal foreign body sensation and the lymphoid hyperplasia in children after tonsillectomy.

Mobility, binding behavior and potential risks of trace metals in the sediments of the fifth largest freshwater lake, China.

The trace metal pollution of sediments in Chaohu Lake, one of the most highly eutrophic lakes in East China, was investigated. Surface sediment (0-5 cm) samples were collected from 35 different positions and analyzed by inductively coupled plasma optical emission spectrometry to determine trace metal contents. Results showed that the mean content of trace metals was as follows: Cr, 85.09 mg kg(-1); Cu, 34.49 mg kg(-1); Ni, 26.46 mg kg(-1); Pb, 34.17 mg kg(-1) and Zn, 107.46 mg kg(-1). The trace metal concentrations from different sampling sites displayed spatial diversity; the heavily polluted sampling sites were close to where estuaries flow in to the lake. A four-step sequential extraction was used to examine the partitioning of the trace metals. Results demonstrated that the percentage of the species bound to the oxidizable phase for all trace metals ranged from 15.6 to 37.7%, while for Cu, Cr and Ni, the main forms were residual (41.3, 62.3 and 69.8%, respectively). Trace metals in the oxidizable fraction may mainly exist in the form of sulfides. The ecological potential risks of trace metals decreased as follows: Pb > Zn > Cu > Cr > Ni.

Chronic resveratrol treatment protects pancreatic islets against oxidative stress in db/db mice.

Resveratrol (RSV) has anti-inflammatory and anti-oxidant actions which may contribute to its cardiovascular protective effects. We examined whether RSV has any beneficial effects on pancreatic islets in db/db mice, an animal model of type 2 diabetes. The db/db and db/dm mice (non-diabetic control) were treated with (db-RSV) or without RSV (db-control) (20 mg/kg daily) for 12 weeks. After performing an intraperitoneal glucose tolerance test and insulin tolerance test, mice were sacrificed, the pancreas was weighed, pancreatic ß-cell mass was quantified by point count method, and the amount of islet fibrosis was determined. 8-Hydroxydeoxyguanosine (8-OHdG), an oxidative stress marker, was determined in 24 h urine and pancreatic islets. RSV treatment significantly improved glucose tolerance at 2 hrs in db/db mice (P = 0.036), but not in db/dm mice (P = 0.623). This was associated with a significant increase in both pancreas weight (P = 0.011) and ß-cell mass (P = 0.016). Islet fibrosis was much less in RSV-treated mice (P = 0.048). RSV treatment also decreased urinary 8-OHdG levels (P = 0.03) and the percentage of islet nuclei that were positive for 8-OHdG immunostaining (P = 0.019). We conclude that RSV treatment improves glucose tolerance, attenuates ß-cell loss, and reduces oxidative stress in type 2 diabetes. These findings suggest that RSV may have a therapeutic implication in the prevention and management of diabetes.

Halogenated organic contaminants (HOCs) in sediment from a highly eutrophicated lake, China: occurrence, distribution and mass inventories.

Halogenated organic contaminants (HOCs) including 16 polybrominated diphenyl ethers (PBDEs) and 37 polychlorinated biphenyls (PCBs) were determined in 49 surfacial sediments from Chaohu Lake, a highly eutrophicated lake, China. PBDEs were detected in almost samples with the range of the total concentration (defined as Σ(16)PBDEs) from 0.84 to 86.6 ng g(-1). Compared with the occurrence of PBDEs in Pearl River Delta and Yangtze River Delta in China, lower percentage of BDE-209 over the concentration of Σ(16)PBDEs was inferred by the high-volume application of penta-BDE mixture product for local domestic furniture purpose. The total concentration of 37 PCBs (Σ(37)PCBs) ranged from 0.05 to 3.36 ng g(-1) with the most detection of PCB-1, -4, -52 and -71. Both the concentrations of Σ(16)PBDE and Σ(37)PCB poorly correlated with total organic carbon (TOC), suggesting the significant contribution of phytoplankton organic carbons to sediment TOC. The contamination by PBDEs and PCBs in western region of the lake was significantly more serious than in eastern lake. Our findings about the higher residues of PBDEs and PCBs in sediments at the estuary of Nanfei River compared to the other estuaries also supported the conclusion that urban area (Hefei city) was the main source of PBDEs and PCBs. The comparison with the concentration of HOC in the present study with those in other lacustrine sediments around the world suggested the contamination by PBDEs in Chaohu Lake is at middle of the global concentration range, whereas PCBs is at low end of the global range which could be elucidated by local economic development and historical usage of PBDEs and PCBs. The mass inventories of HOCs in the lake were estimated at 561 and 38 kg, which corresponds to only 0.000006% and 0.0001% of these global historical produce volumes, respectively.

Spatial distribution and multiple sources of heavy metals in the water of Chaohu Lake, Anhui, China.

In this study, a survey for the spatial distribution of heavy metals in Chaohu Lake of China was conducted. Sixty-two surface water samples were collected from entire lake including three of its main river entrances. This is the first systematic report concerning the content, distribution, and origin of heavy metals (Cu, Cr, Cd, Hg, Zn, and Ni) in the Chaohu Lake water. The results showed that heavy metals (Cu, Cr, Zn, and Ni) concentrations in the estuary of Nanfei River were relatively higher than those in the other areas, while content of Hg is higher in the southeast lake than northwest lake. Moreover, Cd has locally concentration in the surface water from the entire Chaohu Lake. The heavy metal average concentrations, except Hg, were lower than the cutoff values for the first-grade water quality (China Environment Quality Standard) which was set as the highest standard to protect the social nature reserves. The Hg content is between the grades three and four water quality, and other heavy metals contents are higher than background values. The aquatic environment of Chaohu Lake has apparently been contaminated. Both the cluster analysis (CA) and correlation analysis provide information about the origin of heavy metals in the Lake. Our findings indicated that agricultural activities and adjacent plants chimneys may contribute the most to Cd and Hg contamination of Chaohu Lake, respectively.

Resveratrol recruits rat muscle microvasculature via a nitric oxide-dependent mechanism that is blocked by TNFα.

Resveratrol, a polyphenol found in many plants, has antioxidant and anti-inflammatory actions. It also improves endothelial function and may be cardioprotective. Tumor necrosis factor-α (TNFα) causes oxidative stress and microvascular endothelial dysfunction. Whether resveratrol affects microvascular function in vivo and, if so, whether inflammatory cytokines antagonize its microvascular action are not clear. In cultured bovine aortic endothelial cells (BAECs), resveratrol (100 nM) increased the phosphorylation of protein kinase B (Akt), endothelial nitric oxide (NO) synthase (eNOS), and ERK1/2 within 15 min by more than twofold, and this effect lasted for at least 2 h. Treatment of BAECs with TNFα (10 ng/ml) significantly increased the NADPH oxidase activity and the production of hydrogen peroxide and superoxide. Pretreatment of cells with resveratrol (100 nM) prevented each of these. Injection (ip) of resveratrol in rats potently increased muscle microvascular blood volume (MBV; P = 0.007) and flow (MBF; P < 0.02) within 30 min, and this was sustained for at least 2 h. The phosphorylation of Akt in liver or muscle was unchanged. Superimposed systemic infusion of L-NAME (NOS inhibitor) completely abolished resveratrol-induced increases in MBV and MBF. Similarly, systemic infusion of TNFα prevented resveratrol-induced muscle microvascular recruitment. In conclusion, resveratrol activates eNOS and increases muscle microvascular recruitment via an NO-dependent mechanism. Despite the potent antioxidant effect of resveratrol, TNFα at concentrations that block insulin-mediated muscle microvascular recruitment completely neutralized resveratrol's microvascular action. Thus, chronic inflammation, as seen in type 2 diabetes, may limit resveratrol's vasodilatory actions on muscle microvasculature.

Insulin and insulin-like growth factor-I receptors differentially mediate insulin-stimulated adhesion molecule production by endothelial cells.

Patients with type 2 diabetes are hyperinsulinemic and insulin resistant and develop premature atherosclerosis. High concentrations of insulin stimulate the production of adhesion molecules by endothelial cells (ECs). ECs express abundant IGF-I receptors as well as insulin receptors. Whether IGF-I receptors contribute to insulin-induced endothelial production of adhesion molecules is unknown. Bovine aortic ECs (BAECs) were incubated with insulin (100 nm) for 24 h. The cellular content of vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) was measured, and monocyte adhesion to ECs was quantified. Insulin increased both VCAM-1 (P < 0.001) and ICAM-1 (P < 0.0002) content, which was accompanied by an increased number of monocytes adherent to BAECs (P = 0.0001). Inhibition of either MAPK kinase-1 or p38 MAPK but not phosphatidylinositol 3-kinase abolished insulin-mediated production of adhesion molecules. Insulin receptor small interfering RNA knockdown abolished insulin-stimulated increases of ICAM-1 but not VCAM-1. Conversely, IGF-I receptor blockade with either a neutralizing antibody or specific small interfering RNA eliminated insulin-induced VCAM-1 but not ICAM-1 production. Blockade of signaling via either the insulin or IGF-I receptors decreased monocyte adherence to BAECs (P < 0.01 for each). We conclude that insulin and IGF-I receptors differentially mediate the production of adhesion molecules by ECs and monocyte adhesion onto the vascular endothelium in response to the hyperinsulinemic state. Dual-receptor activation may most effectively contribute to the pathogenesis of atherosclerotic disease in diabetes.

Growth hormone exerts acute vascular effects independent of systemic or muscle insulin-like growth factor I.

CONTEXT: Endothelial dysfunction is common in patients with GH deficiency who are at increased risk for premature cardiovascular death. GH regulates vascular tone and reactivity in humans. OBJECTIVE: Our objective was to explore the mechanisms underlying the GH's acute vascular effects. DESIGN AND STUDY SETTING: There were 10 healthy, lean and young, volunteers studied after an overnight fast. GH was infused systemically for 6 h at 0.06 microg/kg.min. Biopsy of the vastus lateralis muscle was done in seven subjects before and after GH infusion. Human aortic endothelial cells (HAECs) were incubated with GH in vitro. RESULTS: GH infusion increased plasma GH to 32.9 +/- 1.5 ng/ml and forearm blood flow by 66% (P < 0.001). GH infusion did not significantly change plasma IGF-I concentrations, muscle IGF-I mRNA expression, and muscle Akt phosphorylation, suggesting a lack of IGF-I action in muscle. Because it was reported that GH exerts an acute vascular effect via a nitric oxide (NO)-dependent mechanism, we performed additional in vitro experiments using HAECs. HAECs express abundant GH receptors. Incubating HAECs with GH at 30 ng/ml for 3 or 6 h did not alter endothelial NO synthase (eNOS) protein content but time dependently increased the phosphorylation and activity of eNOS, thus demonstrating a direct effect of GH on endothelial cells. CONCLUSIONS: GH exerts an acute vascular effect independent of both systemic and local IGF-I production, and this effect is likely via direct action on GH receptors and eNOS in the vascular endothelium.

Activation of p38 mitogen-activated protein kinase abolishes insulin-mediated myocardial protection against ischemia-reperfusion injury.

Myocardial ischemia-reperfusion injury contributes significantly to morbidity and mortality in patients with diabetes. Insulin decreases myocardial infarct size in animals and the rate of apoptosis in cultured cells. Ischemia-reperfusion activates p38 mitogen-activated protein kinase (MAPK), which regulates cellular apoptosis. To examine whether p38 MAPK affects insulin's cardioprotection against ischemia-reperfusion injury, we studied overnight-fasted adult male rats by use of an in vivo rat model of myocardial ischemia-reperfusion. A euglycemic clamp (3 mU.min(-1).kg(-1)) was begun either 10 min before ischemia (InsulinBI), 5 min before reperfusion (InsulinBR), or 30 min after the onset of reperfusion (InsulinAR), and continued until the end of the study. Compared with saline control, insulin decreased the infarct size in both InsulinBI (P < 0.001) and InsulinBR (P < 0.02) rats but not in InsulinAR rats. The ischemic area showed markedly increased phosphorylation of p38 MAPK compared with the nonischemic area in saline animals. Acute activation of p38 MAPK with anisomycin (2 mg/kg iv 10 min before ischemia) had no effect on infarct size in saline rats. However, it completely abolished insulin's protective effect in InsulinBI and InsulinBR rats. Activation of p38 MAPK by anisomycin was associated with marked and persistent elevation in IRS-1 serine phosphorylation. Treatment of animals with SB-239063, a potent and specific inhibitor of p38 MAPK, 10 min before reperfusion enabled insulin-mediated myocardial protection in InsulinAR rats. We conclude that insulin protects myocardium against ischemia-reperfusion injury when given prior to ischemia or reperfusion, and activation of p38 MAPK abolishes insulin's cardioprotective effect.

Tumor necrosis factor-alpha induces insulin resistance in endothelial cells via a p38 mitogen-activated protein kinase-dependent pathway.

Chronic inflammation contributes to vascular insulin resistance and endothelial dysfunction. Systemic infusion of TNF-alpha abrogates insulin's action to enhance skeletal muscle microvascular perfusion. In skeletal muscle TNF-alpha induces insulin resistance via the p38 MAPK pathway. To examine whether p38 MAPK also regulates TNF-alpha-induced vascular insulin resistance, bovine aortic endothelial cells (bAECs) were incubated+/-TNF-alpha (5 ng/ml) for 6 h in the presence or absence of SB203580 (p38 MAPK specific inhibitor, 10 microM) after serum starvation for 10 h. For the last 30 min, cells were treated+/-1 nM insulin, and insulin receptor substrate (IRS)-1, Akt, endothelial nitric oxide synthase (eNOS), p38 MAPK, ERK1/2, c-Jun N-terminal kinase, and AMP-activated protein kinase (AMPK) phosphorylation, and eNOS activity were measured. TNF-alpha increased p38 MAPK phosphorylation, potently stimulated IRS-1 serine phosphorylation, and blunted insulin-stimulated IRS-1 tyrosine and Akt phosphorylation and eNOS activity. TNF-alpha also potently stimulated the phosphorylation of ERK1/2 and AMPK. Treatment with SB203580 decreased p38 MAPK phosphorylation back to the baseline and restored insulin sensitivity of IRS-1 tyrosine and Akt phosphorylation and eNOS activity in TNF-alpha-treated bAECs without affecting TNF-alpha-induced ERK1/2 and AMPK phosphorylation. We conclude that in cultured bAECs, TNF-alpha induces insulin resistance in the phosphatidylinositol 3-kinase/Akt/eNOS pathway via a p38 MAPK-dependent mechanism and enhances ERK1/2 and AMPK phosphorylation independent of the p38 MAPK pathway. This differential modulation of TNF-alpha's actions by p38 MAPK suggests that p38 MAPK plays a key role in TNF-alpha-mediated vascular insulin resistance and may contribute to the generalized endothelial dysfunction seen in type 2 diabetes mellitus and the cardiometabolic syndrome.

The vascular endothelial cell mediates insulin transport into skeletal muscle.

The pathways by which insulin exits the vasculature to muscle interstitium have not been characterized. In the present study, we infused FITC-labeled insulin to trace morphologically (using confocal immunohistochemical methods) insulin transport into rat skeletal muscle. We biopsied rectus muscle at 0, 10, 30, and 60 min after beginning a continuous (10 mU x min(-1) x kg(-1)), intravenous FITC-insulin infusion (with euglycemia maintained). The FITC-insulin distribution was compared with that of insulin receptors (IR), IGF-I receptors (IGF-IR), and caveolin-1 (a protein marker for caveolae) in skeletal muscle vasculature. We observed that muscle endothelium stained strongly for FITC-insulin within 10 min, and this persisted to 60 min. Endothelium stained more strongly for FITC-insulin than any other cellular elements in muscle. IR, IGF-IR, and caveolin-1 were also detected immunohistochemically in muscle endothelial cells. We further compared their intracellular distribution with that of FITC-insulin in cultured bovine aortic endothelial cells (bAECs). Considerable colocalization of IR or IGF-IR with FITC-insulin was noted. There was some but less overlap of IR or IGF-IR or FITC-insulin with caveolin-1. Immunoprecipitation of IR coprecipitated caveolin-1, and conversely the precipitation of caveolin-1 brought down IR. Furthermore, insulin increased the tyrosine phosphorylation of caveolin-1, and filipin (which inhibits caveolae formation) blocked insulin uptake. Finally, the ability of insulin, IGF-I, and IGF-I-blocking antibody to diminish insulin transport across bAECs grown on transwell plates suggested that IGF-IR, in addition to IR, can also mediate transendothelial insulin transit. We conclude that in vivo endothelial cells rapidly take up and concentrate insulin relative to plasma and muscle interstitium and that IGF-IR, like IR, may mediate insulin transit through endothelial cells in a process involving caveolae.

Insulin at physiological concentrations selectively activates insulin but not insulin-like growth factor I (IGF-I) or insulin/IGF-I hybrid receptors in endothelial cells.

In muscle, physiologic hyperinsulinemia, presumably acting on endothelial cells (ECs), dilates arterioles and regulates both total blood flow and capillary recruitment, which in turn influences glucose disposal. In cultured ECs, however, supraphysiological (e.g. >or=10 nM) insulin concentrations are typically used to study insulin receptor (IR) signaling pathways and nitric oxide generation. IGF-I receptors (IGF-IRs) are more abundant than IR in ECs, and they also respond to high concentrations of insulin. To address whether IR mediates responses to physiologic insulin stimuli, we examined the insulin concentration dependence of IR and IGF-IR-mediated insulin signaling in bovine aortic ECs (bAECs). We also assessed whether insulin/IGF-I hybrid receptors were present in bAECs. Insulin, at 100-500 pM, significantly stimulated the phosphorylation of IRbeta, Akt1, endothelial isoform of nitric oxide synthase, and ERK 1/2 but not the IGF-IRbeta subunit. At concentrations 1-5 nm or greater, insulin dose-dependently enhanced the tyrosine phosphorylation of IGF-IRbeta, and this was inhibited by IGF-IR neutralizing antibody. In addition, immunoprecipitation of IRbeta pulled down the IGF-IRbeta, and the IRbeta immunocytochemically colocalized with IGF-IRbeta, suggesting that ECs have insulin/IGF-I hybrid receptors. We conclude that: 1) insulin at physiological concentrations selectively activates IR signaling in bAECs; 2) bAECs express IGF-IR and insulin/IGF-I hybrid receptors in addition to IR; 3) high concentrations of insulin (>or=1-5 nM) activate IGF-IR and hybrid receptors as well as IR; and 4) this crossover activation can confound interpretation of studies of insulin action in ECs when high insulin concentrations are used.