Laboratory Investigation of Skid Resistance for Steel Slag Utilization as Chip Seal

Slag as waste material of steel-making process has similar characteristics with aggregate that has been widely used in pavement construction. The use of slag as chip seal aggregate to provide skid resistance needs to be analyzed. In this laboratory study, the chip seal samples are made using steel slag and natural aggregate. The bonding materials used are asphalt and epoxy resin. Skid resistance tests for all chip seal samples and also hot rolled sheet pavement without chip seal application are performed using the Portable British Pendulum Tester. The results show the variations of chip seal aggregate weight are inconsistent. The natural aggregate used as chip seal material could produce high skid resistance value of 10.3% higher than that using steel slag. Also the skid resistance of chip seal with the ALD 3 mm are not significantly different with that of ALD 6 mm. Similar results occur on the skid resistance of chip seals using epoxy resin and asphalt

GAPDH overexpression increases in SNO-Hsp60, but not SNO-DHRS2 levels.

<p>SNO-Hsp60 (<b>A</b>) and SNO-DHRS2 (<b>B</b>) levels from Hep2G cells transfected with either a control GFP plasmid or siRNA scramble, a plasmid encoding DDK-tagged GAPDH for overexpression, a GAPDH siRNA for knock-down, or a plasmid encoding DDK-tagged GAPDH_C150S were assessed via SNO-RAC proteomic analysis, followed by label-free peptide quantification (n = 6).</p

Myocardial perfusion protocols.

<p>Perfusion protocols for <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0111448#pone-0111448-g002" target="_blank">Figure 2</a> (<b>A</b>) and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0111448#pone-0111448-t001" target="_blank">Table 1</a> (<b>B</b>).</p

HepG2 cell line as a model system for examining GAPDH as a trans-S-nitrosylase.

<p>(<b>A</b>) Expression of neuronal and endothelial isoforms of NO synthase in HepG2 cells. Representative western blots are shown for neuronal (top right) and endothelial (top left) NO synthase and β-actin (lower). (<b>B</b>) Mitochondrial GAPDH protein levels were assessed after the addition of purified GAPDH to isolated HepG2 mitochondria. Representative western blots for GAPDH (upper), TOM20 (center), and the α subunit of F₁F₀-ATPase (lower) in HepG2 mitochondria. Control: non-treated mitochondrial control; GAPDH: purified GAPDH treated mitochondria; (n = 3). (<b>C</b>) and (<b>D</b>) Hep2G cells were transfected with either a control GFP plasmid or siRNA scramble, a plasmid encoding DDK-tagged GAPDH for overexpression, a GAPDH siRNA for knock-down, or a plasmid encoding DDK-tagged GAPDH_C150S for overexpression. Representative western blots are shown together with the densitometry of GAPDH normalized to β-actin for total GAPDH (upper; GAPDH, GAPDH-DDK, GAPDH_C150S-DDK) and β-actin (lower; *p<0.05 vs. control; n = 6).</p

Mitochondrial GAPDH and total GAPDH levels following cardioprotection.

<p>(<b>A</b>) GAPDH protein levels in the mitochondrial fraction of hearts subjected to control or an IPC-I/R protocol were assessed via liquid chromatography-tandem mass spectrometry, followed by label-free peptide quantification (*p<0.05 vs. control, n = 3). (<b>B</b>) Total GAPDH protein levels in hearts subjected to control or an IPC-I/R protocol were assessed via western blot analysis. Representative western blots are shown for total GAPDH (upper) and enolase (lower) in whole heart homogenates together with the densitometry of GAPDH normalized to enolase (n = 3).</p

SNO proteins from HepG2 cells as identified via SNO-RAC proteomic analysis.

<p>SNO proteins from HepG2 cells as identified via SNO-RAC proteomic analysis.</p

SNO-GAPDH increases mitochondrial SNO levels.

<p>A representative gel is shown for mitochondrial SNO levels measured via DyLight800 fluorescence after the addition of purified GAPDH or SNO-GAPDH. MW: molecular weight marker; GSNO (+): GSNO treatment was used as a positive control for mitochondrial SNO; Control: non-treated mitochondrial control; GSNO: filtered GSNO treated mitochondria; GAPDH: purified GAPDH treated mitochondria; SNO-GAPDH: SNO-GAPDH treated mitochondria. <i>Please note</i>: the filtration procedure serves to remove excess GSNO following the incubation of purified GAPDH with GSNO (n = 3).</p

GAPDH and SNO-GAPDH are imported into the matrix of heart mitochondria.

<p>Mitochondrial GAPDH protein levels were assessed after the addition of purified GAPDH or SNO-GAPDH to isolated mitochondria from control and IPC hearts. (<b>A</b>) Representative western blots for GAPDH (upper), TOM20 (center), and the α subunit of F₁F₀-ATPase (lower) in control heart and IPC heart mitochondria. Control: non-treated mitochondrial control; GAPDH: purified GAPDH treated mitochondria; SNO-GAPDH: SNO-GAPDH treated mitochondria. (<b>B</b>) GAPDH import into control and IPC heart mitochondria as assessed via the percentage of GAPDH following trypsin digestion compared to GAPDH levels prior to the addition of trypsin. GAPDH levels were assessed via densitometry (n = 3).</p

GAPDH acts as a mitochondrial trans-S-nitrosylase.

<p>GAPDH and SNO-GAPDH can be imported into the mitochondria, where SNO-GAPDH targets Hsp60, ACAT1, and VDAC1 as a mitochondrial trans-<i>S</i>-nitrosylase.</p

GAPDH binding partners in cytosolic and mitochondrial fractions of hearts subjected to control or GSNO perfusion.

<p>GAPDH binding partners in cytosolic and mitochondrial fractions of hearts subjected to control or GSNO perfusion.</p