196 research outputs found
The Effects Of Rapport-Building Style on Childrenās Reports of a Staged Event
Three- to 9-year-old children (N = 144) interacted with a photographer and were interviewed about the event either a week or a month later. The informativeness and accuracy of information provided following either open-ended or direct rapport building were compared. Children in the open-ended rapport-building condition provided more accurate reports than children in the direct rapport-building condition after both short and long delays. Open-ended rapport-building led the 3- to 4-year-olds to report more errors in response to the first recall question about the event, but they went on to provide more accurate reports in the rest of the interview than counterparts in the direct rapport-building condition. These results suggest that forensic interviewers should attempt to establish rapport with children using an open-ended style
COMPASS CRUSADERS
Grade Level(s): 6-12The purpose of this lesson is to learn how to use a compass and a G.P.S. unit AND how to use that knowledge in the field while applying orienteering and mathematical skills.Westlane Middle School, Indianapolis, I
Preparation and technical considerations for percutaneous cannulation for veno-arterial extracorporeal membrane oxygenation.
BACKGROUND: The most frequent limb complications from peripheral veno-arterial extracorporeal membrane oxygenation (VA-ECMO) are limb ischemia and localized bleeding. To minimize these risks, perfusion of the distal limb with peripheral percutaneous cannulation was done.
TECHNIQUE: Percutaneous cannulation with a distal perfusion port was performed in all patients. During the VA-ECMO, distal limb perfusion was monitored using near-infrared spectroscopy to assess tissue oxygenation. At the decannulation, patch angioplasty was performed to prevent the development of narrowing of the artery at the cannulation site.
CONCLUSIONS: Using our standard technique, we have not experienced any limb loss related to ischemia or bleeding
Modified Appleby Procedure with Arterial Reconstruction for Locally Advanced Pancreatic Adenocarcinoma: A Literature Review and Report of Three Unusual Cases.
BACKGROUND: Pancreatic body and tail ductal adenocarcinomas are often diagnosed with local vascular invasion of the celiac axis (CA) and its various branches. With such involvement, these tumors have traditionally been considered unresectable. The modified Appleby procedure allows for margin negative resection of some such locally advanced tumors. This procedure involves distal pancreatectomy with en bloc splenectomy and CA resection and relies on the presence of collateral arterial circulation via an intact pancreaticoduodenal arcade and the gastroduodenal artery to maintain prograde hepatic arterial perfusion. When the resultant collateral circulation is inadequate to provide sufficient hepatic and gastric arterial inflow, arterial reconstruction (AR) is necessary to supercharge the inflow. Herein, we review all reported cases of AR with modified Appleby procedures that we have identified in the literature, and we report our experience of three recent cases with arterial reconstruction including two cases with arterial bypasses not requiring interposition grafting.
METHODS: Perioperative and oncologic outcomes from our Institutional Review Board-approved database of pancreatic resections at the Thomas Jefferson University were reviewed. Additionally, PubMed search for cases of distal or total pancreatectomy with celiac axis resection and concurrent AR was performed.
RESULTS: From the literature, 12 reports involving 28 patients were identified of distal and total pancreatectomy with AR after CA resection. The most common AR in the literature, performed in 12 patients, was a bypass from the aorta to the common hepatic artery (CHA) using a variety of interposition conduits. In our institutional experience, patient #1 had a primary side-to-end aorto-CHA bypass, patient #2 had a primary end-to-end bypass of the transected distal CHA to the left gastric artery in the setting a replaced left hepatic artery, and patient #3 required an aortic to proper hepatic artery bypass with saphenous vein graft and portal venous reconstruction. All patients recovered from their operations without ischemic complications, and they are currently 16, 15, and 13 months post-op, respectively.
CONCLUSIONS: The criteria for resectability in patients with locally advanced pancreatic body and tail neoplasms are expanding due to increasing experience with AR in the setting of the modified Appleby procedure. When performing AR, primary arterial re-anastomosis may be considered preferable to interposition grafting as it decreases the potential for the infectious and thrombotic complications associated with conduits and it reduces the number of vascular anastomoses from two to one. Consideration must also be given to normal variant anatomy of the hepatic circulation during operative planning as the origin of the left gastric artery is resected with the CA. The modified Appleby procedure with AR, when used in appropriately selected patients, offers the potential for safe, margin negative resection of locally advanced pancreatic body and tail tumors
Entropic and enthalpic components of catalysis in the mutase and lyase activities of Pseudomonas aeruginosa PchB
This document is the Accepted Manuscript version of a Published Work that appeared in final form in the Journal of the American Chemical Society, copyright Ā© American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see http://doi.org/10.1021/ja202091a.The isochorismate-pyruvate lyase from Pseudomonas aeruginosa (PchB) catalyzes two pericyclic reactions, demonstrating the eponymous activity and also chorismate mutase activity. The thermodynamic parameters for these enzyme-catalyzed activities, as well as the uncatalyzed isochorismate decomposition, are reported from temperature dependence of kcat and kuncat data. The entropic effects do not contribute to enzyme catalysis as expected from previously reported chorismate mutase data. Indeed, an entropic penalty for the enzyme-catalyzed mutase reaction (ĪSā” = -12.1 Ā± 0.6 cal/molK) is comparable to that of the previously reported uncatalyzed reaction, whereas that of the enzyme-catalyzed lyase reaction (ĪSā” = -24.3 Ā± 0.6 cal/molK) is larger than that of the uncatalyzed lyase reaction (-15.77 Ā± 0.02 cal/molK) documented here. With the assumption that chemistry is rate-limiting, we propose that a reactive substrate conformation is formed upon loop closure of the active site and that ordering of the loop contributes to the entropic penalty for converting the enzyme substrate complex to the transition state
Modification of residue 42 of the active site loop with a lysinemimetic sidechain rescues isochorismate-pyruvate lyase activity in Pseudomonas aeruginosa PchB
PchB is an isochorismate-pyruvate lyase from Pseudomonas aeruginosa. A positively charged lysine residue is located in a flexible loop that behaves as a lid to the active site, and the lysine residue is required for efficient production of salicylate. A variant of PchB that lacks the lysine at residue 42 has a reduced catalytic free energy of activation of up to 4.4 kcal/mol. Construction of a lysine isosteric residue bearing a positive charge at the appropriate position leads to the recovery of 2.5ā2.7 kcal/mol (about 60%) of the 4.4 kcal/mol by chemical rescue. Exogenous addition of ethylamine to the K42A variant leads to a neglible recovery of activity (0.180 kcal/mol, roughly 7% rescue), whereas addition of propylamine caused an additional modest loss in catalytic power (0.056 kcal/mol, or ā2% loss). This is consistent with the view that (a) the lysine-42 residue is required in a specific conformation to stabilize the transition state and (b) the correct conformation is achieved for a lysine-mimetic sidechain at site 42 in the course of loop closure, as expected for transition-state stabilization by the side chain ammonio-function. That the positive charge is the main effector of transition state stabilization is shown by the construction of a lysine-isosteric residue capable of exerting steric effects and hydrogen bonding but not electrostatic effects, leading to a modest increase of catalytic power (0.267 ā 0.505 kcal/mol of catalytic free energy, or roughly 6 ā 11% rescue)
Redesign of MST enzymes to target lyase activity instead promotes mutase and dehydratase activities
The isochorismate and salicylate synthases are members of the MST family of enzymes. The isochorismate synthases establish an equilibrium for the conversion chorismate to isochorismate and the reverse reaction. The salicylate synthases convert chorismate to salicylate with an isochorismate intermediate; therefore, the salicylate synthases perform isochorismate synthase and isochorismate-pyruvate lyase activities sequentially. While the active site residues are highly conserved, there are two sites that show trends for lyase-activity and lyase-deficiency. Using steady state kinetics and HPLC progress curves, we tested the āinterchangeā hypothesis that interconversion of the amino acids at these sites would promote lyase activity in the isochorismate synthases and remove lyase activity from the salicylate synthases. An alternative, āpermuteā hypothesis, that chorismate-utilizing enzymes are designed to permute the substrate into a variety of products and tampering with the active site may lead to identification of adventitious activities, is tested by more sensitive NMR time course experiments. The latter hypothesis held true. The variant enzymes predominantly catalyzed chorismate mutase-prephenate dehydratase activities, sequentially generating prephenate and phenylpyruvate, augmenting previously debated (mutase) or undocumented (dehydratase) adventitious activities
Lysine221 is the general base residue of the isochorismate synthase from Pseudomonas aeruginosa (PchA) in a reaction that is diffusion limited
The isochorismate synthase from Pseudomonas aeruginosa (PchA) catalyzes the conversion of chorismate to isochorismate, which is subsequently converted by a second enzyme (PchB) to salicylate for incorporation into the salicylate-capped siderophore pyochelin. PchA is a member of the MST family of enzymes, which includes the structurally homologous isochorismate synthases from E. coli (EntC and MenF) and salicylate synthases from Yersinia enterocolitica (Irp9) and Mycobacterium tuberculosis (MbtI). The latter enzymes generate isochorismate as an intermediate before generating salicylate and pyruvate. General acid ā general base catalysis has been proposed for isochorismate synthesis in all five enzymes, but the residues required for the isomerization are a matter of debate, with both lysine221 and glutamate313 proposed as the general base (PchA numbering). This work includes a classical characterization of PchA with steady state kinetic analysis, solvent kinetic isotope effect analysis and by measuring the effect of viscosogens on catalysis. The results suggest that isochorismate production from chorismate by the MST enzymes is the result of general acid ā general base catalysis with a lysine as the base and a glutamic acid as the acid, in reverse protonation states. Chemistry is determined to not be rate limiting, favoring the hypothesis of a conformational or binding step as the slow step
Physical condition and stress levels during early development reflect feeding rates and predict pre- and post-fledging survival in a nearshore seabird
The effects of acute environmental stressors on reproduction in wildlife are often difficult to measure because of the labour and disturbance involved in collecting accurate reproductive data. Stress hormones represent a promising option for assessing the effects of environmental perturbations on altricial young; however, it is necessary first to establish how stress levels are affected by environmental conditions during development and whether elevated stress results in reduced survival and recruitment rates. In birds, the stress hormone corticosterone is deposited in feathers during the entire period of feather growth, making it an integrated measure of background stress levels during development. We tested the utility of feather corticosterone levels in 3- to 4-week-old nestling brown pelicans (Pelecanus occidentalis) for predicting survival rates at both the individual and colony levels. We also assessed the relationship of feather corticosterone to nestling body condition and rates of energy delivery to nestlings. Chicks with higher body condition and lower corticosterone levels were more likely to fledge and to be resighted after fledging, whereas those with lower body condition and higher corticosterone levels were less likely to fledge or be resighted after fledging. Feather corticosterone was also associated with intracolony differences in survival between ground and elevated nest sites. Colony-wide, mean feather corticosterone predicted nest productivity, chick survival and post-fledging dispersal more effectively than did body condition, although these relationships were strongest before fledglings dispersed away from the colony. Both reproductive success and nestling corticosterone were strongly related to nutritional conditions, particularly meal delivery rates. We conclude that feather corticosterone is a powerful predictor of reproductive success and could provide a useful metric for rapidly assessing the effects of changes in environmental conditions, provided pre-existing baseline variation is monitored and understood
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