NMR studies of caldesmon-calmodulin interactions

The binding of the calcium-regulatory protein calmodulin (CaM) to caldesmon (CaD) contributes to the regulation of smooth muscle contraction. Two regions of caldesmon have been identified as putative calmodulin-binding domains. We have earlier reported on the binding of one of these domains to calmodulin (Zhang & Vogel (1994) Biochemistry 33, 1163-1171). Here we have studied the binding of CaM to synthetic peptides of CaD which contain: (1) both the first and second CaM-binding domains; (2) the second CaM-binding domain; and (3) the sequence between the first and second CaM-binding domains. Two-dimensional transferred nuclear Overhauser enhancement proton NMR measurements as well as circular dichroism studies of a 22-residue peptide NKETAGLKVGVSSRINEWLTK, which contains the second CaM-binding domain, show that only the C-terminal half of the peptide becomes alpha-helical upon binding to CaM. Somewhat surprisingly, the shorter 9-residue peptide SRINEWLTK was sufficient to form a 1:1 complex with CaM; this peptide appears to bind as a 3(10)-helix. Proton-carbon-13 correlation NMR titration studies with specifically labeled [methyl-13C]methionine CaM were used to study the participation of the hydrophobic regions in both domains of the dumbbell shaped CaM in peptide binding. Binding of a 54-residue CaD peptide containing both CaM-binding domains affects all the 8 Met residues in the two hydrophobic domains of CaM (only Met 76 in the linker region of CaM is not involved), while binding of the second CaM-binding domain of CaD influences principally Met 51, 71, and Met 124, 144. Simultaneous binding to CaM of two peptides comprising the first and the second CaM-binding domains also caused changes to all Met residues except Met 76. Taken together, these data demonstrate that both CaM-binding domains of CaD can bind simultaneously to the two hydrophobic regions of CaM.     

Effect of metabolic acidosis on insulin action and secretion in uremia

BACKGROUND: Metabolic acidosis affects both vitamin D and insulin metabolism. Vitamin D is important in modulation of both insulin secretion and insulin sensitivity in uremia. The present study examines the effect of correction of metabolic acidosis on insulin action and secretion as well as 1,25 vitamin D3 concentrations in uremic patients. METHODS: Eight patients (age 18 +/- 1 year) on maintenance hemodialysis with metabolic acidosis were studied before and after two weeks of oral sodium bicarbonate (NaHCO3) treatment to correct the acidosis. To control for the effect of additional sodium, they were also studied after two weeks of an equivalent amount of oral sodium chloride (NaCl). Controls consisted of 7 healthy controls (age 19 +/- 1 year). Insulin sensitivity was measured by the hyperinsulinemic euglycemic clamp technique. Insulin secretion was measured by the hyperglycemic clamp technique. RESULTS: Oral NaHCO3 treatment led to significant increases in venous pH and serum bicarbonate concentrations but no significant change in intact parathyroid hormone (PTH) concentrations. Circulating 1,25 dihydroxyvitamin [(OH)2] D3 were significantly lower than control values initially and increased significantly after treatment. Oral NaCl did not change any of the biochemical parameters. Before treatment of acidosis, uremic patients had lower insulin sensitivity (insulin resistance) during constant hyperinsulinemia and lower insulin secretion during constant hyperglycemia compared with controls. Following two weeks of NaHCO3 treatment there were significant increases in insulin sensitivity and insulin secretion, although the values did not normalize. There were no changes in insulin sensitivity or insulin secretion following two weeks of NaCl. CONCLUSION: Treatment of metabolic acidosis increased both insulin sensitivity and insulin secretion in patients with uremia. This was accompanied by an increase in the circulating levels of 1,25(OH)2D3 but no change in those of parathyroid hormone.     

Polyhedral studies for minimum-span graph labelling with integer distance constraints

This paper studies the polytope of the minimum‐span graph labelling problems with integer distance constraints (DC‐MSGL). We first introduce a few classes of new valid inequalities for the DC‐MSGL defined on general graphs and briefly discuss the separation problems of some of these inequalities. These are the initial steps of a branch‐and‐cut algorithm for solving the DC‐MSGL. Following that, we present our polyhedral results on the dimension of the DC‐MSGL polytope, and that some of the inequalities are facet defining, under reasonable conditions, for the polytope of the DC‐MSGL on triangular graphs.     

Clock Faults Induced Min and Max Delay Violations

In this paper, we show that clock faults producing duty-cycle variations, which have been proven very likely, can give rise to min or max delay violations. This mandates new, specific testing approaches for clock faults, to avoid them to compromise the system correct operation in the field, with dramatic effects on product quality and defect level. We then introduce a new scheme that can be employed to detect the clock faults causing duty-cycle variations.     

Using a real-time integrated communication system to monitor the progress and quality of construction works

Site monitoring is an indispensable procedure in construction quality control. It does not only minimize construction defects and human errors but also supports project team members making strategic decisions at critical points throughout the construction stages. This paper presents a cost-effective construction site monitoring system integrating a long-range wireless network, network cameras, and a web-based collaborative platform. The system supports simultaneous user access therefore project team members could view real-time captured images or video of a construction site, discuss and exchange ideas with gadgets such as video conference, text and shared whiteboard at a distance via the Internet. It was carefully configured in order to maintain the reliability under the reactive conditions of the construction sites. The system has been implemented and tested on two construction sites and promising results were obtained.     

Nanovesicles Based on Mixtures of a Biantennary Glycolipid with Ionic Co‐Surfactants

A biantennary surfactant based on a synthetic C₁₆‐maltoside was chosen to prepare vesicles for a potential vesicular drug delivery system. The synthesis comprised of three stages: Initial synthesis of β‐d ‐maltose octaacetate was followed by glycosidation of 2‐hexyl‐decanol and final glycolipid deacetylation. Both α‐ and β‐anomers were prepared and their anomeric purity was evaluated by ¹H NMR. Owing to the low water solubility of the glycolipid, addition of ionic co‐surfactants was believed to promote the surfactant distribution, thus leading to smaller and more uniform vesicles. The assembly behavior of the surfactant systems was studied by contact penetration under an optical polarizing microscope, while interfacial properties were determined by surface tension measurements. Vesicles were prepared by injection of an ethanolic solution into bulk water and investigated by dynamic light scattering and field emission scanning electron microscopy. Contact of the surfactant mixtures with water indicated a high tendency to exhibit the lamellar phase and confirmed the expected low molecular solubility. These findings suggest a potential of the surfactant to form stable vesicles. Injection of an ethanolic surfactant solution into bulk water gave sub‐micrometer sized vesicles with a narrow size distribution. Application of ionic co‐surfactants reduced the vesicle size. In particular ～20 % of anionic SDS proved highly effective, lowering the vesicle size by nearly one decade, thus accessing nano‐sized vesicles. Encapsulation of a water‐soluble drug was achieved in a 76 ± 10 % efficiency.     

The effects of long‐term exposure of flax fiber reinforced polymer to salt solution at high temperature on tensile properties

Most research on natural fiber composites has been primarily conducted on randomly‐oriented fibers. This study is focused on the short‐ and long‐term performances of flax fiber‐reinforced polymer (Flax‐FRP) made from continuous unidirectional fiber mats, and compares it to Glass‐FRP composite. The study looked into the effect of number of layers on properties, comparing wet layup (WL) to vacuum bag (VB) molding, and aging in a 3.5% salt solution for up to 365 days at 23, 40, and 55°C. Results show that Flax‐FRP has a tensile strength and modulus of one third the values of Glass‐FRP. Using the VB process, Flax‐FRP showed a strength and modulus 18 and 36% higher, respectively, than WL specimens. As the number of layers increased from one to five, the strength and modulus also increased but stabilized at three layers. After 365 days of conditioning at 23, 40, and 55°C, WL specimens showed a strength retention of 81, 73, and 69%, respectively. Using the Arrhenius relationship, it was estimated that both WL and VB Flax‐FRP would retain 60% of their tensile strength after 100 years of saltwater exposure at an annual mean temperature of 10°C. POLYM. COMPOS., 37:3234–3244, 2016. © 2015 Society of Plastics Engineers     

Taylor vortex flow in presence of internal baffles

We present a study on Taylor vortex flow in between a rotating inner cylinder and a stationary outer cylinder with vertical or horizontal baffles. A mineral oil is used as the working fluid in the annular channel of 60 mm in height and 11.6 mm in gap width. The flow pattern in the annulus is investigated by particle image velocimetry (PIV) and computational fluid dynamics (CFD). Both the experimental and simulation results show that the presence of vertical baffles influences the vertical positions of vortices—it is primarily significant for the bottom vortex. Increasing number of vertical baffles gradually decreases the azimuthal variation in the vertical position. The baffles also lead to the occurrence of a circulation flow immediately upstream and downstream each baffle plate in the radial–azimuthal plane. On the other hand, an annular horizontal baffle with sufficient width alters the vortex structure in the gap, for instance, fewer vortices are observed. With two horizontal baffles in the annulus, the flow pattern is largely affected by the baffle width and the distance between the two baffles. This study provides a better understanding of Taylor vortex in presence of internal baffles, which will be important for practical applications of Taylor vortex devices.