Folding mechanism of the alpha-subunit of tryptophan synthase, an alpha/beta barrel protein: global analysis highlights the interconversion of multiple native, intermediate, and unfolded forms through parallel channels

A variety of techniques have been used to investigate the urea-induced kinetic folding mechanism of the alpha-subunit of tryptophan synthase from Escherichia coli. A distinctive property of this 29 kDa alpha/beta barrel protein is the presence of two stable equilibrium intermediates, populated at approximately 3 and 5 M urea. The refolding process displays multiple kinetic phases whose lifetimes span the submillisecond to greater than 100 s time scale; unfolding studies yield two relaxation times on the order of 10-100 s. In an effort to understand the populations and structural properties of both the stable and transient intermediates, stopped-flow, manual-mixing, and equilibrium circular dichroism data were globally fit to various kinetic models. Refolding and unfolding experiments from various initial urea concentrations as well as forward and reverse double-jump experiments were critical for model discrimination. The simplest kinetic model that is consistent with all of the available data involves four slowly interconverting unfolded forms that collapse within 5 ms to a marginally stable intermediate with significant secondary structure. This early intermediate is an off-pathway species that must unfold to populate a set of four on-pathway intermediates that correspond to the 3 M urea equilibrium intermediate. Reequilibrations among these conformers act as rate-limiting steps in folding for a majority of the population. A fraction of the native conformation appears in less than 1 s at 25 degrees C, demonstrating that even large proteins can rapidly traverse a complex energy surface.     

Small signal frequency response studies for plasma electrolytic oxidation

Plasma assisted electrochemical treatments provide new possibilities in surface modification of various materials including light weight alloys. However, their large-scale application is still restricted, mainly due to poor understanding of the process mechanisms and consequent limitations in process control and automation. This problem can be resolved if the frequency response (FR) of the system is known and applied for process diagnostics. Our previous work has shown the effectiveness of the large signal mode for FR studies of plasma electrolytic oxidation (PEO) of aluminium. This research is dedicated to FR measurements during PEO of Al in the small signal mode corresponding to small perturbations of voltage signal around large DC values. The study was carried out during PEO of Al at DC voltages which were varied from 450 to 600 V. The FR obtained is a frequency dependent admittance of the PEO electrolyser; this complex number is represented by a modulus and a phase angle. Under potentiostatic conditions, the modulus evolution strongly correlates with the average current value; therefore, it bears insufficient amounts of independent information. The FR phase angle measured within this study was never obtained before. Depending on the frequency, it varies between 0° and 70 ° in the capacitive domain. One of the most notable features of this characteristic is low values at 500-5000 Hz when microdischarges appear during PEO. The other feature is a correlation with the coating growth. As a result, a new diagnostic tool was developed and shown to be effective for evaluation of microdischarges and surface properties during the treatment, thus decreasing the uncertainty in the system.     

Validation of caregiver interviews to diagnose common causes of severe neonatal illness

The objective of this study was to validate retrospective caregiver interviews for diagnosing major causes of severe neonatal illness and death. A convenience sample of 149 infants aged < 28 days with one or more suspected diagnoses of interest (low birthweight/severe malnutrition, preterm birth, birth asphyxia, birth trauma, neonatal tetanus, pneumonia, meningitis, septicaemia, diarrhoea, congenital malformation or injury) was taken from patients admitted to two hospitals in Dhaka, Bangladesh. Study paediatricians performed a standardised history and physical examination and ordered laboratory and radiographic tests according to study criteria. With a median interval of 64.5 days after death or hospital discharge, caregivers of 118 (79%) infants were interviewed about their child's illness. Using reference diagnoses based on predefined clinical and laboratory criteria, the sensitivity and specificity of particular combinations of signs (algorithms) reported by the caregivers were ascertained. Sufficient numbers of children with five reference standard diagnoses were studied to validate caregiver reports. Algorithms with sensitivity and specificity > 80% were identified for neonatal tetanus, low birthweight/severe malnutrition and preterm delivery. Algorithms with specificities > 80% for birth asphyxia and pneumonia had sensitivities < 70%, or alternatively had high sensitivity with lower specificity. In settings with limited access to medical care, retrospective caregiver interviews provide a valid means of diagnosing several of the most common causes of severe neonatal illness and death.     

The PingER project: active Internet performance monitoring for theHENP community

The extraordinary network challenges presented by high energy nuclear and particle physics experiments has created a need for network monitoring both to understand present performance and to allocate resources to optimize performance between laboratories, and the universities and institutes collaborating on present and future experiments. The resulting Internet end-to-end performance monitoring project is called PingER. The monitoring infrastructure reflects the wide geographical spread of the collaborations, and involves a large number of research and commercial networks. The architecture of the data acquisition and methodology of the analysis have evolved over several years, and are described here in their present state. The strengths and weaknesses of the project are reviewed, and the derived metrics are discussed in terms of their diagnostic functions. The observed short-term effects and long-term trends are reviewed, and plans for future developments are described     

What's in a manner of speaking? Children's sensitivity to partner-specific referential precedents.

Do young children form “referential pacts”? If a person has referred to an object with a certain term (e.g., the horse), will children expect this person to use this term in the future but allow others to use a different expression (e.g., the pony)? One hundred twenty-eight children between 3 and 5 years old co-operated with an experimenter (E1) to move toys to new locations on a shelf. E1 established referential terms for all toys in a warm-up game. Then, either the original partner, E1, or a new partner, E2, played a second game with the same toys. In this game, the experimenters referred to toys using either their original terms from the warm-up game or new terms. Children were slower to react to new terms than old, and this difference in reaction times was greater in the original partner condition (but only on the first trial). Children sometimes protested at the use of new terms, doing so regardless of their interlocutor's identity. We contrast these findings with those for adults and discuss their implications for the debate regarding the nature of referential pacts. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Association between socioeconomic status and metabolic syndrome in women: Testing the reserve capacity model.

Objective: Low socioeconomic status (SES) environments may impede the development of a bank of resources, labeled reserve capacity, and may also be stressful, thereby depleting available reserves. In consequence, lower SES persons may experience more negative emotions, leading to adverse health consequences. The authors tested the reserve capacity model in relation to the metabolic syndrome. Design: There were 401 initially healthy women who followed longitudinally for 12 years. Self-reported characteristics, stressors, and cardiovascular risk factors were measured repeatedly. Structural equation modeling was used to evaluate hypothesized relationships. Main Outcome Measure: Metabolic syndrome factor. Results: Confirmatory factor analysis verified reserve capacity as the aggregate of optimism, self-esteem, and social support, and negative emotion as the aggregate of depressive symptoms, anger, and tension. Structural equation modeling showed two pathways to the metabolic syndrome factor, (χ2(59) = 111.729, p ldf = 1.894; CFI = .956; RMSEA = .047): direct from low SES to the metabolic syndrome factor (B = -0.19, t = -3.24, p = .001); and indirect, from low SES to low reserve capacity to high negative emotions to the metabolic syndrome factor (B = -0.024, t = -2.05, p = .04). Conclusion: Low SES may increase risk for metabolic syndrome, in part, through reserve capacity and negative emotions. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Clinical experience of trabeculotomy for the surgical treatment of aniridic glaucoma

OBJECTIVE: The purpose of this study is to determine the efficacy of initial trabeculotomy in the patient with aniridic glaucoma. DESIGN: Clinical charts were reviewed. PARTICIPANTS: Twenty-nine eyes of 16 patients with aniridia were studied. INTERVENTION: Glaucoma surgery was performed. As an initial procedure, trabeculotomy was performed in 12 eyes, other surgery was performed in 17 eyes (trabeculectomy, 5; goniotomy, 5; other, 7). MAIN OUTCOME MEASURES: Success was defined as an intraocular pressure (IOP) of 21 mmHg or lower, and no further surgery was performed. RESULTS: Ten (83%) of 12 eyes obtained IOP control after first (6 eyes) or second (4 eyes) trabeculotomy with a mean follow-up period of 9.5 years. Five eyes maintained visual acuity of 20/40 to 20/200. No serious complications were found after trabeculotomy. Three (18%) of 17 eyes were controlled with the first glaucoma surgery other than trabeculotomy (goniotomy, trabeculectomy, trabeculectomy combined with trabeculotomy, and Molteno implant). Good IOP control was obtained in 8 (47%) of 17 eyes after several surgeries with a mean follow-up period of 10.4 years. Four of 17 eyes became phthisical. CONCLUSION: This study suggests that trabeculotomy is the preferred initial operation for uncontrolled glaucoma with aniridia.     

Rearrangement of Epoxides to Allylic Alcohols in the Presence of Reusable Basic Resins

We have modified Merrifield’s resin to provide polymers containing secondary amine groups. Lithiation of the solids gives strongly basic yet poorly nucleophilic resins useful for rearrangement of epoxides to their corresponding allylic alcohols. The resins are easy to handle, non-volatile, non-toxic, and are easily recovered and reused, providing environmental and economic benefits that might have commercial viability.