Chill activation of compatible solute transporters in Corynebacterium glutamicum at the level of transport activity

The gram-positive soil bacterium Corynebacterium glutamicum harbors four osmoregulated secondary uptake systems for compatible solutes, BetP, EctP, LcoP, and ProP. When reconstituted in proteoliposomes, BetP was shown to sense hyperosmotic conditions via the increase in luminal K(+) and to respond by instant activation. To study further putative ways of stimulus perception and signal transduction, we have investigated the responses of EctP, LcoP, and BetP, all belonging to the betaine-carnitine-choline transporter family, to chill stress at the level of activity. When fully activated by hyperosmotic stress, they showed the expected increase of activity at increasing temperature. In the absence of osmotic stress, EctP was not activated by chill and LcoP to only a very low extent, whereas BetP was significantly stimulated at low temperature. BetP was maximally activated at 10 degrees C, reaching the same transport rate as that observed under hyperosmotic conditions at this temperature. A role of cytoplasmic K(+) in chill-dependent activation of BetP was ruled out, since (i) the cytoplasmic K(+) concentration did not change significantly at lower temperatures and (ii) a mutant BetP lacking the C-terminal 25 amino acids, which was previously shown to have lost the ability to be activated by luminal K(+), was fully competent in chill sensing. When heterologously expressed in Escherichia coli, BetP did not respond to chill stress. This may indicate that the membrane in which BetP is inserted plays an important role in chill activation and thus in signal transduction by BetP, different from the previously established K(+)-mediated process.     

Geometric limit of multiple local Limberg flaps: a flap design.

The Limberg rhombic flap is a reliable and widely used technique in head and neck surgery. Since Limberg introduced his original design in 1946, several modifications of the technique have been described. Although a single Limberg flap is frequently used at the face to close small to medium defects, multi-Limberg flap techniques can help the surgeon to cover moderate to large defects of the extremities, trunk, and back. In this study, a design of four neighboring local Limberg flaps to cover a moderate to large defect without using a skin graft is introduced. It is believed that this design is the geometric limit of multiple Limberg flaps that can entirely cover a single large rhombic defect, because one Limberg flap unit can only be adjoined by three others, one from the tip and two from the sides. This flap design of four local Limberg flaps is also the only geometrically possible design that can keep all the bases of these four flaps free of incisions if one attempts to prepare four small Limberg flaps around a large rhombic defect.     

Pre and postoperative assessment of regional cerebral blood flow in hydrocephalus by 99mTc-hexamethyl-propylenamine oxime SPECT

Cerebrovascular changes resulting from hydrocephalus still remain to be investigated. It has been suggested that hydrocephalus distorts the large feeding arteries and that the collapse of capillaries results in decreased cerebral blood flow. This clinical study was designed to evaluate the effect of shunting on regional cerebral blood flow in patients with obstructive hydrocephalus of varying duration. Technetium-99m hexamethyl propyleamine oxime (99mTc-HMPAO) was used to measure the cerebral perfusion, semiquantitatively, since the pattern of its distribution in brain is somewhat similar to that of regional cerebral blood flow (rCBF), and the pre and postoperative semiquantitative rCBF values of each lobe were calculated. Fifteen patients (8 F, 7 M) underwent both CT and single photon emission tomography (SPECT) using 99mTc-HMPAO examination before and 1 week after shunting. Mean percentage of all lobes were calculated by subtracting the preoperative mean rCBF of all lobes from the corresponding postoperative values. The patients were classified into 3 groups according to the mean percentage of all lobes. Group A: showed a marked increase in mean cortical blood flow (+ 16.00 +/- 2.9%), group B: a moderate increase (11.27 +/- 4.8%), and in group C: there was the least improvement in mean cortical blood flow (+ 1.17 +/- 2.7%). The mean duration of hydrocephalus of group A, group B and group C was 5 +/- 0.5 weeks, 8 +/- 1 weeks and more than 12 weeks, respectively. Psychological testing and clinical observation of the daily activities of the patients postoperatively showed some correlation with increased rCBF and clinical improvement.(ABSTRACT TRUNCATED AT 250 WORDS)     

Thiol antioxidants protect human lens epithelial (HLE B-3) cells against tert-butyl hydroperoxide-induced oxidative damage and cytotoxicity

Oxidative damage to lens epithelial cells plays an important role in the development of age-related cataract, and the health of the lens has important implications for overall ocular health. As a result, there is a need for effective therapeutic agents that prevent oxidative damage to the lens. Thiol antioxidants such as tiopronin or N-(2-mercaptopropionyl)glycine (MPG), N-acetylcysteine amide (NACA), N-acetylcysteine (NAC), and exogenous glutathione (GSH) may be promising candidates for this purpose, but their ability to protect lens epithelial cells is not well understood. The effectiveness of these compounds was compared by exposing human lens epithelial cells (HLE B-3) to the chemical oxidant tert-butyl hydroperoxide (tBHP) and treating the cells with each of the antioxidant compounds. MTT cell viability, apoptosis, reactive oxygen species (ROS), and levels of intracellular GSH, the most important antioxidant in the lens, were measured after treatment. All four compounds provided some degree of protection against tBHP-induced oxidative stress and cytotoxicity. Cells treated with NACA exhibited the highest viability after exposure to tBHP, as well as decreased ROS and increased intracellular GSH. Exogenous GSH also preserved viability and increased intracellular GSH levels. MPG scavenged significant amounts of ROS, and NAC increased intracellular GSH levels. Our results suggest that both scavenging ROS and increasing GSH may be necessary for effective protection of lens epithelial cells. Further, the compounds tested may be useful for the development of therapeutic strategies that aim to prevent oxidative damage to the lens.