Residues essential for the function of SecE, a membrane component of the Escherichia coli secretion apparatus, are located in a conserved cytoplasmic region

Protein export in Escherichia coli is absolutely dependent on two integral membrane proteins, SecY and SecE. Previous deletion mutagenesis of the secE gene showed that only the third of three membrane-spanning segments and a portion of the second cytoplasmic region are necessary for its function in protein export. Here we further define the residues important for SecE function. Alignment of the SecE homologues of various eubacteria reveals that they all contain one membrane-spanning segment, compared with three in E. coli SecE, and that the most conserved region among them lies in their putative cytoplasmic amino termini; little homology exists in their membrane-spanning segments. The SecE homologue of the extreme thermophilic bacterium Thermotoga maritima was cloned and found to complement a deletion of secE in E. coli. Deletion or replacement of the cytoplasmic region of E. coli SecE eliminated SecE function, indicating that this sequence is essential for a functional secretion machinery. Mutant analysis suggests that the most important function of the third membrane-spanning segment is to maintain the proper topological arrangement of the conserved cytoplasmic domain.     

Differential contractile response of cultured microvascular pericytes to vasoactive agents

OBJECTIVE: Microvascular pericytes may contract in two different ways: In the first, a circumferential or radial mechanical force applied at right angles to the long axis of the vessel may constrict the underlying vessel affecting blood flow and transmural pressure. Retraction and elongation of pericyte processes may also occur tangentially and at right angles to the vessel axis and alter microvessel permeability by changing the amount of ablumenal surface covered or the openness of interendothelial junctions. In this study, cultured pericytes were utilized as a model experimental system to determine if vasoactive stimulation changes their shape in a manner consistent with this hypothesis. METHODS: Pericytes cultured from isolated rate capillaries were subjected to angiotensin II and histamine. Their response was monitored by measuring the area of non-yielding substrate covered by the pericytes and the manner in which their shape changed. Shape changes were quantified by calculating the surface area: perimeter perimeter ratios. RESULTS: Histamine significantly reduced surface area covered and the surface area:perimeter ratio. The pericyte processes retracted, resulting in elongated, spindle-shaped cells. These effects were nullified by the H1 blocker diphenhydramine suggesting a receptor-specific response. Angiotensin II also elicited contraction and reduced surface area, but the cells contracted laterally and longitudinally. The surface area: perimeter ratios also decreased. CONCLUSIONS: These results indicate that pericytes are capable of two types of contractile responses in culture, depending on the specific vasoactive stimulus.     

Reduction and uptake of methylene blue from rat air spaces

The use of methylene blue (MB) to estimate dilution of epithelial lining fluid, which occurs during bronchoalveolar lavage (BAL), is complicated by loss of this redox dye from the air spaces. The rate of MB uptake from the air spaces of isolated rat lungs and the effects of oxidation and reduction on this process were investigated in this study. Movement of MB from the air spaces to perfusate was compared with the corresponding transport of 125I-labeled albumin, [14C]-dextran, 99mTc-labeled diethylenetriaminepentaacetate, [3H]-sucrose, and 3H2O. By the end of 2 min, MB concentrations in the BAL had fallen by 58 +/- 4% (SE; n = 11) and 3H2O by 78 +/- 2% (n = 13), whereas concentrations of the other indicators decreased by approximately 6%. All but 10% of the 3H2O lost from the air spaces was found in the perfusate, whereas 19% of the lost MB was not recovered in the perfusate, suggesting retention of MB in the pulmonary tissues. Absorption of MB from the air spaces was slowed by 20% when the lungs were left unperfused, and absorption was accelerated threefold by reduction of MB to leukomethylene blue with Na2S2O4. In contrast, MB losses from the air space were slowed by the oxidizing agent K3Fe(CN)6 and by addition of superoxide dismutase or ascorbic oxidase. It is therefore possible that ascorbic acid and O2- entering the air spaces reduce MB to the uncharged leuko form. Lowering the pH of the BAL fluid to 3.5 also slowed MB reabsorption. This suggests that acid aspiration may stimulate release of oxidants into the air spaces.     

Clinical trial of a new lightwand device (Trachlight) to intubate the trachea

BACKGROUND: Transillumination of the soft tissue of the neck using a lighted stylet (lightwand) is an effective and safe intubating technique. A newly designed lightwand (Trachlight) incorporates modifications to improve the brightness of the light source as well as flexibility. The goal of this study was to determine the effectiveness and safety of this device in intubating the trachea of elective surgical patients. METHODS: Healthy surgical patients were studied. Patients with known or potential problems with intubation were excluded. During general anesthesia, the tracheas were intubated randomly using either the Trachlight or the laryngoscope. Failure to intubate was defined as lack of successful intubation after three attempts. The duration of each attempt was recorded as the time from insertion of the device into the oropharynx to the time of its removal. The total time to intubation (TTI), an overall measure of the ease of intubation, was defined as the sum of the durations of all (as many as three) intubation attempts. Complications, such as mucosal bleeding, lacerations, dental injury, and sore throat, were recorded. RESULTS: Nine hundred fifty patients (479 in the Trachlight group and 471 in the laryngoscope group) were studied. There was a 1% failure rate with the Trachlight, and 92% of intubations were successful on the first attempt, compared with a 3% failure rate and an 89% success rate on the first attempt with the laryngoscope (P not significant). All failures were followed by successful intubation using the alternate device. The TTI was significantly less with the Trachlight compared with the laryngoscope (15.7 +/- 10.8 vs. 19.6 +/- 23.7 s). For laryngoscopic intubation, the TTI was longer for patients with limited mandibular protrusion and mentohyoid distance, with a larger circumference of the neck, and with a high classification according to Mallampatti et al. However, there was no relation between the TTI and any of the airway parameters for Trachlight. There were significantly fewer traumatic events in the Trachlight group than in the laryngoscope group (10 vs. 37). More patients complained of sore throat in the laryngoscope group than in the Trachlight group (25.3% vs. 17.1%). CONCLUSIONS: In contrast to laryngoscopy, the ease of intubation using the Trachlight does not appear to be influenced by anatomic variations of the upper airway. Intubation occasionally failed with the Trachlight but in all cases was resolved with direct laryngoscopy. The failures of direct laryngoscopy were resolved with Trachlight. Thus the combined technique was 100% successful in intubating the tracheas of all patients.     

Children's Physiological Indices of Empathy and Their Socioemotional Adjustment: Does Caregivers' Expressivity Matter?

Relations of heart rate and skin conductance reactions to mildly evocative empathy-inducing slides with socioemotional functioning were examined for 154 children (mean age = 9 years, 5 months). In addition, maternal expressivity was tested as a moderator of these relations. Parents and teachers rated children's socioemotional functioning, and a behavioral measure of children's regulation was obtained. Boys who exhibited higher skin conductance and higher heart rate to slides depicting negative emotions were better regulated, less emotionally intense, and better adjusted than their peers. Furthermore, boys' regulation and adjustment were positively related to such physiological responding to negative slides if maternal negative expressivity was relatively low or moderate, but not high. Fewer findings were obtained for girls or for positive slides. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Behavior and movements of Indian meal moths (Plodia interpunctella Hübner) during commodity infestation

A model warehouse with carefully regulated environmental conditions was used to study the behavior of adult Indian meal moths during commodity infestation. Sequences of moth distributions in the warehouse, calculated by spatial analysis, clearly showed the coordinated movements of moth populations from eclosion to death. Many facets of adult behavior in a warehouse were coordinated with the photoperiod. Adult moths emerged at the end of a photophase and fairly rapidly moved to the walls and, to a lesser extent, to the undersides of the commodity pallets. Most females are mated in the first 24 h after emergence, largely during the scotophase. Air circulation within the warehouse probably compromised pheromone-directed guidance of males to females, although pheromones may still have a major role in regulating other aspects of male mating behavior. When mating subsided, many of the males flew upwind to the air circulating unit, but only during the photophases. The females moved to the return air side of the warehouse and then migrated towards the same end as the males, but only during the scotophases. They moved from pallet to pallet largely by walking on the warehouse floor and laid eggs in or on the cups of commodity. Direct oviposition on the commodity during the infestation period was lower than expected, probably because commodity odors were dissipated by air circulation and did not provide appropriate orientation. At 144 h after emergence, 90% of the moth population had died.     

The State of Disequilibrium between U and Th Series Isotopes in Marine Sediments from the Sudanese Red Sea Coast

The geochemical behavior of U, Th series nuclides in surface marine sediments collected fromthe Red Sea coastal environment of Sudan was examined using daughter/parent and nonisotopic activity ratios. Pronounced anomalous behavior of several series of nuclides has been observed, particularly at shallower depths. On the average, the ²³⁴U/²³⁸U ratio is fairly constant and close to 1.13, suggesting scavenging of ²³⁴U from the overlying water column. In shallow-water sediments collected from the biologically productive area of the fringing reef and from the tidal-flat of Sanganeb atoll, compared to harbor and deep-sea sediments, ²³⁸U was found in large excess relative to ²³²Th. The ²²⁸Th/²³²Th disequilibrium indicates rapid sedimentation at shallower sites, which agrees well with data reported in the literature for similar environments. Depletion was observed of precursor thorium isotopes (²³⁰Th, ²³²Th) with respect to their mobile daughter isotopes of radium (²²⁶Ra, ²²⁸Ra). On the other hand, there is surface enrichment of ²²⁸Th and ²¹⁰Po with respect to their progenitors ²²⁸Ra and ²²⁶Ra, as demonstrated by the ²²⁸Th : ²²⁸Ra and ²¹⁰Po : ²²⁶Ra activity ratio, which is greater than unity in all the sediments analyzed. The ²¹⁰Po/²²⁶Ra ratio is fairly constant (average 3.2).     

Porosity distribution in highly compressible cake: Experimental and theoretical verification of the dense skin

The rate of filtration and the water content of cake are influenced by the existence of a dense skin in a highly compressible cake. The phenomenon of the dense skin has been rarely studied, and its existence has not been verified experimentally. In this study, the porosity variation in a very compressible cake is measured by using a new experimental apparatus, and with this the existence of dense skin has been established experimentally. ‘Unified theory on solid-liquid separation’, a recently developed theory, is utilized for calculating the porosity variation in a very compressible cake.     

Relating matrix metalloproteinase structure to function: why the "hemopexin" domain?

Matrix metalloproteinases are thought to be key players in the remodelling activity of cells associated with both physiological and pathological processes. They share a relatively conserved structure with a number of identifiable modules linked to their specific functions. The structure of the individual domains of a number of matrix metalloproteinases have now been elucidated. Here we review these data in the light of complementary studies on the behaviour of these enzymes in biological systems. In particular we focus on the C-terminal hemopexin-like domain which has intriguingly specific roles in individual matrix metalloproteinases.