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Photochromism of diarylethenes with two nitronyl nitroxides: photoswitching of an intramolecular magnetic interaction.

Photochromic diarylethenes that have p-phenylene-substituted benzothiophene aryl groups with and without nitronyl nitroxide radicals at both ends of the molecules were synthesized. The absorption maxima of the closed-ring isomers showed a hypsochromic shift with the increase in the pi-conjugated chain length. The unique behavior was attributed to the stabilization by the resonant quinoid structures. Both photocyclization and photocycloreversion quantum yields of the diarylethene with nitronyl nitroxide radicals were found to increase with the increase in the pi-conjugated chain length. Photoswitching of the magnetic interaction between two nitronyl nitroxide radicals was studied by means of ESR spectroscopy. The change in exchange interaction between open- and closed-ring isomers of 1,2-bis[6-[4-[4-(1-oxyl-3-oxide-4,4,5,5-tetramethylimidazolin-2-yl)phenyl]phenyl]-2-methyl-1-benzothiophen-3-yl]hexafluorocyclopentene was determined to be more than 30-fold.

Journal Article↗

Interpersonal violence in prehistoric San Pedro de Atacama, Chile: behavioral implications of environmental stress.

The prehistoric population of San Pedro de Atacama lived through periods marked by prosperity and interregional interaction, as well as times of severe drought, social stress, and widespread poverty. A sample of 682 crania was analyzed for evidence of cranial trauma in order to assess changing patterns of interpersonal violence during the occupation of the oasis. It was hypothesized that the level of traumatic injuries in this population would parallel some of the changes seen in the archaeological record. Low fracture rates would be expected in periods of affluence and environmental stability, while periods characterized by environmental extremes and state collapse would yield elevated rates of aggression. This analysis found that rates of trauma escalated from 5.1% (5/99) in the earliest period, to 10.9% (10/92) in the Middle Horizon (AD 600-950). Although it may reflect problems related to increasing population density in the oasis, this increase is surprising, given that the early period witnessed the shift to permanent settlements, and the middle period was one of prosperity and plentiful resource availability. Trauma rates peaked at 35.6% (16/45) in an early Late Intermediate period (AD 950-1400) cemetery, with other Late Intermediate cemeteries demonstrating similarly high rates of traumatic injury. The elevated trauma rates during this period correlate with major droughts, the concentration of settlements on the oasis' east side, fortified structures, and material poverty, all reflected in the archaeological record. As the Late Intermediate waned and environmental conditions improved, trauma concomitantly decreased (7.0%), and remained low throughout the Inka occupation (AD 1400-1532). This indicates that while the Atacama was not peaceful, violence became commonplace only during periods of great social change and resource stress.

Adult↗

Efficient C-F and C-C activation by a novel N-heterocyclic carbene-nickel(0) complex.

The NHC-stabilized complex [Ni2(iPr2Im)4(cod)] (1) was isolated in good yield from the reaction of [Ni(cod)2] with 1,3-diisopropylimidazole-2-ylidene (iPr2Im). Compound 1 is a source of the [Ni(iPr2Im)2] complex fragment in stoichiometric and catalytic transformations. The reactions of 1 with ethylene and CO under atmospheric pressure or with equimolar amounts of diphenylacetylene lead to the compounds [Ni(iPr2Im)2(eta2-C2H4)] (2), [Ni(iPr2Im)2(eta2-C2Ph2)] (3), and [Ni(iPr2Im)2(CO)2] (4) in good yields. In all cases the [Ni(iPr2Im)2] complex fragment is readily transferred without decomposition or fragmentation. In the infrared spectrum of carbonyl complex 4, the CO stretching frequencies are observed at 1847 and 1921 cm(-1), and are significantly shifted to lower wavenumbers compared with other nickel(0) carbonyl complexes of the type [NiL2(CO)2]. Complex 1 activates the C--F bond of hexafluorobenzene very efficiently to give [Ni(iPr2Im)2(F)(C6F5)] (5). Furthermore, [Ni2(iPr2Im)4(cod)] (1) is also an excellent catalyst for the catalytic insertion of diphenylacetylene into the 2,2' bond of biphenylene. The reaction of 1 with equimolar amounts of biphenylene at low temperature leads to [Ni(iPr2Im)2(2,2'-biphenyl)] (6), which is formed by insertion into the strained 2,2' bond. The reaction of diphenylacetylene and biphenylene at 80 degrees C in the presence of 2 mol % of 1 as catalyst yields diphenylphenanthrene quantitatively and is complete within 30 minutes.

Journal Article↗

Analysis of 3-hydroxydodecanedioic acid for studies of fatty acid metabolic disorders: preparation of stable isotope standards.

Current diagnostic tests to detect disorders of fatty acids metabolism, such as long-chain hydroxyacyl CoA dehydrogenase deficiency (LCHAD), are hampered by insensitivity or a long delay time required for results. Children with LCHAD deficiency are known to excrete 3-hydroxydicarboxylic acids with chain lengths of 10-16 carbons, but a quantitative method to measure excretion of these potentially diagnostically important compounds has not been reported. We report synthetic schemes for synthesis of 3-hydroxydodecanedioic acid and a di-deuterated analog, suitable for use in a stable-isotope dilution mass spectrometric analytical approach. Evaluation of several common derivatization protocols to produce a volatile derivative for gas chromatography determined that trimethylsyl derivatives produced the best efficiency and stability. Positive-ion chemical ionization mass spectrometry provided the greatest yield of characteristic ions. These results indicate the basic reagents needed to develop sensitive and accurate 3-hydroxydodecanedioic acid measurements for diagnosis of LCHAD deficiency and other fatty acid oxidation disorders.

3-Hydroxyacyl CoA Dehydrogenases↗

Modeling the role of disulfide bonds in protein folding: entropic barriers and pathways.

The role of disulfide bonds in directing protein folding is studied using lattice models. We find that the stability and the specificity of the disulfide bond interactions play quite different roles in the folding process: Under some conditions, the stability decreases the overall rate of folding; the specificity, however, by yielding a simpler connectivity of intermediates, always increases the rate of folding. This conclusion is intimately related to the selection mechanism entailed by entropic driving forces, such as the loop formation probability, and entropic barriers separating the native and the many native-like metastable states. The folding time is found to be a minimum for a certain range of the effective disulfide bond interaction. Examination of a model, which allows for the formation of disulfide bonded intermediates, suggests that folding proceeds via a three-stage multiple pathways kinetics. We show that there are pathways to the native state involving only native-like intermediates, as well as those that are mediated by nonnative intermediates. These findings are interpreted in terms of the appropriate energy landscape describing the barriers connecting low energy conformations. The consistency of our conclusions with several experimental studies is also discussed.

Computer Simulation↗

Determination of chlorpyrifos and its metabolites in rat brain tissue using coupled-column liquid chromatography/electrospray ionization tandem mass spectrometry.

A method has been developed to quantify chlorpyrifos (O,O-diethyl-O-[3,5,6,-trichloro-2-pyridyl] phosphorothionate) and its metabolites chlorpyrifos-oxon (O,O-diethyl-O-[3,5,6,trichloro-2-pyridinyl] phosphate) and TCP (3,5,6,-trichloro-2-pyridinol) in rat brain tissue by coupled-column liquid chromatography/electrospray ionization tandem mass spectrometry (LC/LC/ESI-MS/MS). Rat brains were homogenized and treated by protein precipitation using ice-cold acetonitrile. The supernatant was directly injected onto the coupled-column system. Sample clean-up was achieved on a Zorbax Extend-C(18) column (2.1 x 50 mm, 5 microm) using a mobile phase of acetonitrile/water with 0.0025% formic acid (40:60, v/v). The compounds were separated isocratically on a Zorbax Eclipse XDB C(8) column (2.0 x 150 mm, 5 microm) using a mobile phase of acetonitrile/water with 0.0025% formic acid (75:25, v/v). Chlorpyrifos and chlorpyrifos-oxon were detected in positive ion mode using multiple reaction monitoring (MRM). TCP was detected in negative ion mode using precursor-to-precursor transition monitoring. The method was validated and the specificity, linearity, limit of quantitation (LOQ), precision, accuracy, stability, and recoveries were determined. Calibration curves for all three analytes yielded correlation coefficients of 0.993 or greater. The LOQs were 25.3 ng/g for chlorpyrifos and 6.3 ng/g for chlorpyrifos-oxon and TCP. All precision relative standard deviations (RSDs) were less than 16% for the LOQ and less than 11% for the other QC samples. This method was successfully applied to six rats that were injected subcutaneously with chlorpyrifos.

Animals↗

A family of low and high copy replicative, integrative and single-stranded S. cerevisiae/E. coli shuttle vectors.

We describe a set of replicative, integrative and single-stranded shuttle vectors constructed from the pUC19 plasmid that we use routinely in our experiments. They bear a yeast selectable marker: URA3, TRP1 or LEU2. Replicative vectors carrying different yeast replication origins have been constructed in order to have plasmids based on the same construction with a high or low copy number per cell and with different mitotic stabilities. All the vectors are small in size, provide a high yield in Escherichia coli and efficiently transform Saccharomyces cerevisiae. These plasmids have many of the unique sites of the pUC19 multicloning region and many of them allow for the screening of plasmids with an insert by alpha-complementation. The nucleotide sequence of each of them is completely known.

Base Sequence↗

Thermodynamics of cross links.

Comparison of the melting temperatures of native lysozyme and a cross-linked ester derivative of lysozyme (Imoto and Rupley, 1973) yielded a value of 5.5 kcal/mole for the free energy of stabilization developed through forming the cross link and a value of zero for the corresponding enthalpy. There is close agreement between experiment and calculation from the statistical theory of polymers.

Calorimetry↗

Features of cholera and Vibrio parahaemolyticus diarrhoea endemicity in Calabar, Nigeria.

The clinical and epidemiological features of acute vibrio diarrhoeal disease were studied in 881 patients seen at the University of Calabar Teaching Hospital (UCTH), Calabar, Nigeria, between January and December 1989. Stools and rectal swabs of patients and randomly-selected control subjects were microscopically and culturally examined for the presence of enteric pathogens. Households of vibrio diarrhoea cases and matched controls were visited for ecologic studies. Of a total of 108 (12.3%) culturally-confirmed bacterial diarrhoeas, 47 (43.5%) were due to Escherichia coli, 33 (30.6%) to Vibrio cholerae-01 (classical and El Tor biotypes) and V. parahaemolyticus, while shigellae and salmonellae accounted for 29 (26.9%) and 9 (8.3%) cases, respectively. Most cholera case households clustered within the ancient neighbourhood of the inner city, characterized by poorly developed water and sewage disposal systems. A preponderance of vibrio diarrhoea patients were children < or = 10 years. Adult cases involved mostly females. The only case of diarrhoea-related death involved an eight-month old child with kwashiorkor and V. parahaemolyticus infection. Incidence of vibrio diarrhoeas was seasonal, with most cases occurring during the dry season followed by subsidence at the onset of rainy season. Bimodal peaks of vibrio diarrhoeal episodes observed over the period appeared to coincide with periods of acute water scarcity, high temperature, increased fishing activities and trade traffic on the Calabar River estuary. Of the environments sampled, only clam shells from a case household and river sediments yielded vibrio pathogens on culture. Ecological factors that are capable of stabilizing a focus of vibrio diarrhoea endemicity in this area are highlighted.

Adolescent↗

A reliable method for the use of oligonucleotides as probes in blot-hybridization experiments.

We have developed a ligation and specific-primer radiolabeling method that allows the use of oligonucleotides as probes in blot-hybridization experiments. The major advantage of the protocol is that standard hybridization and washing conditions may be used and yield high signals and low background. The observed increase in the stability and intensity of the hybridization signals appears to result from both increased length and specific radioactivity of the hybridization probe.

Base Sequence↗

Expression of functional hexahistidine-tagged ricin B in tobacco.

Ricin B (RTB), the lectin subunit of ricin, shows promise as an effective mucosal adjuvant and carrier for use in humans. In order to obtain a recombinant plant source of RTB that is devoid of the toxic ricin A subunit, we expressed RTB in Nicotiana tabacum. RTB was engineered with an N-terminal hexahistidine tag (His-RTB), which may affect protein stability. Lactose-affinity purification of His-RTB from leaves yielded three major glycosylated products of 32, 33.5 and 35 kDa. Their identity as RTB was verified by mass spectrometry and immunoblotting with anti-ricin antibodies. Functionality of His-RTB was confirmed by binding to asialofetuin, lactose and galactose.

Gene Expression↗

Catheter optimization during contrast ventriculography: a prospective randomized trial.

Although newer contrast mediums have improved hemodynamic stability during left ventriculography, the image quality and hence the diagnostic yield of the procedure is largely determined by the choice of catheter and injection technique. In this study 125 patients were prospectively assigned at random to undergo ventriculography using either of two pigtail catheters, one with a straight shaft throughout its length (straight) and one with a 145-degree bend placed 6.5 cm from the distal end (angled). Injectate composition, flow rate, and volume were held constant for all injections, and baseline clinical and catheterization variables were found to be similar in each group. The angled catheter exhibited a statistically superior ease of insertion (p = 0.038) and took less time to position (p = 0.012), saving a mean of 23 seconds of fluoroscopy time per procedure. It was also associated with superior contour edge definition (p = 0.037) and a trend toward more uniform distribution of contrast medium (p = 0.089). Compared with the straight catheter, the angled catheter was less frequently accompanied by artifactual mitral regurgitation (p = 0.038) but was equally likely to provoke ventricular arrhythmias during injection. These observed differences may be explained in part by the tendency for angled catheters to more frequently localize in the central as opposed to the inferoposterior region of the left ventricular cavity (mean distance from center = 0.53 vs 1.10 cm, respectively; p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗

1-Aminocyclopropane-1-carboxylic acid as a substrate of peroxidase: conditions for oxygen consumption, hydroperoxide generation and ethylene production.

Conditions in which 1-aminocyclopropane-1-carboxylic acid (ACC) functions as a substrate of peroxidase have been investigated by measuring oxygen consumption in the reaction medium and the production of ethylene. In both cases, the presence of Mn2+ and either H2O2 or the activated form of peroxidase, namely compound I of peroxidase, was found to be essential. Both oxygen consumption and ethylene production were dependent on enzyme concentration, the optimum ACC/Mn2+ ratio being 1:1. Oxygen consumption in a system with ACC, Mn2+ and compound I showed an enzyme-dependent lag phase and then proceeded to total depletion, suggesting that the system itself generates hydroperoxides that completed the catalytic cycle of the enzyme. The presence of these hydroperoxides in the reaction medium was detected by a colorimetric method. High H2O2 concentration progressively decreased oxygen consumption, the same effect being produced by catalase. Ethylene production was oxygen dependent, mediated by ACC-free radicals and gave a poor yield. The results suggest that the fate of these ACC-free radicals determines the yield in ethylene. These radicals must be oxidized immediately, otherwise their stabilization to hydroperoxides would prevent ethylene production.

Amino Acids↗

Renaturation of glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides after denaturation in 4 M guanidine hydrochloride: kinetics of aggregation and reactivation.

In 4 M guanidine hydrochloride (GdnHCl), the dimeric enzyme glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides (G6PD) dissociated to subunits and was extensively unfolded. Rapid dilution of this high GdnHCl concentration allowed G6PD to partially renature, as measured by enzyme reactivation, to a level which depended on the conditions employed. The fraction of the enzyme which did not renature aggregated and precipitated out of solution, a process which could not be substantially prevented by stabilizing additives. Based on the enzyme concentration dependence of the reactivation yield and on a comparison of the aggregation and reactivation rates, it was determined that aggregation and reactivation compete kinetically for a partially-folded intermediate only very early in the process, during the rapid GdnHCl-dilution step. The kinetics of G6PD reactivation were sigmoidal, indicating that this process involves more than one rate-limiting reaction. The kinetics depended on enzyme concentration in a higher than first-order manner, indicating that association of subunits is one of the rate-limiting reactions. A renaturation mechanism compatible with these observations is described, which involves a bi-unimolecular (subunit association-folding) reaction sequence, with rate constants equal to 2.19 microM-1 min-1 and 0.140 min-1, respectively. This mechanism involves an inactive, dimeric, G6PD-folding intermediate, a species whose existence has recently been established by equilibrium denaturation experiments (Plomer, J.J. and Gafni, A. (1992) Biochim. Biophys. Acta 1122, 234-242).

Buffers↗

Use of palladium(II) chloride as colour-forming reagent in determination of pralidoxime chloride in water and tablets.

Pralidoxime chloride (PAM-2Cl) has been determined spectrophotometrically in Britton-Robinson buffer solution at pH = 6.45; the method is based on measurement of the absorbance of the Pd(II)-pralidoxime complex at 327 nm. Studies of the composition of the complex by Job's continuous variation method, the molar ratio method and Bent-French's method yielded a Pd(II):pralidoxime ratio of 1:1. The conditional stability constant (K') of the complex at the optimum pH of 6.45 and an ionic strength (mu) of 0.3 M was found to be 10(5.2). The molar absorptivity was 1.05 x 10(4) 1 mol(-1) cm(-1). Beer's law was obeyed at concentrations up to 60 microM. The detection limit was 0.55 microg ml(-1). The relative standard deviation (N = 10) was 0.28-1.03%. The method was accurate and sensitive for the analysis of PAM-2Cl in water and tablets.

Journal Article↗

The development and validation of the impact of multiple sclerosis scale and the symptoms of multiple sclerosis scale.

OBJECTIVE: To develop and validate the Impact of Multiple Sclerosis Scale (IMSS) and the Symptoms of Multiple Sclerosis Scale (SMSS) using the Extended Disability Status Scale (EDSS) for construct validity. DESIGN: Panel design involving test-retest over 4 months. SETTING: A mailed survey. PARTICIPANTS: Volunteers with a diagnosis of multiple sclerosis (MS) recruited from an MS support service in Australia: 193 people (mean age, 39y) and 150 people participated at time 1 and time 2, respectively. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Principal components analyses, the Cronbach alpha, and descriptive statistics for the 2 scales; correlations for construct validity with the EDSS and retest; and confirmatory factor analysis to test the stability of IMSS and SMSS components over time. RESULTS: The IMSS yielded 5 independent and reliable components; the SMSS yielded 3 components; both component structures were stable over time. These scales showed convergent validity with the EDSS. CONCLUSIONS: The IMSS and SMSS are psychometrically sound scales suitable for clinical and research purposes to assess the symptoms and impact of MS.

Adult↗

Towards the protein phosphatase-based biosensor for microcystin detection.

A colorimetric test for the detection of microcystins based on immobilised protein phosphatase (PP) has been developed. A PP2A produced by molecular engineering has been used and its performance has been compared to those of commercial PP2A and PP1. Covalent immobilisation of the enzyme using glutaraldehyde, encapsulation by sol-gel and entrapment with photocrosslinkable poly(vinyl alcohol) bearing styrylpyridinium groups (PVA-SbQ) have been compared, the latter method providing the highest immobilisation yields. Screen-printed carbon electrodes (SPEs), Maxisorp microtiter wells and Ultrabind modified polyethersulfone affinity membranes have been used as immobilisation supports. Whilst the highest immobilisation yields were obtained with microtiter wells, the highest operational and storage stabilities were achieved with carbon SPEs and membranes, respectively. The immobilisation of PP by PVA-SbQ provided a means to preserve the enzymatic activity, which decreased at fast rates when the enzyme was kept in solution. The colorimetric test using p-nitrophenyl phosphate has demonstrated that the immobilised enzyme is able to recognise both microcystin variants (MC-LR and MC-RR), although optimisation work should be performed to achieve appropriate limits of detection. With the purpose to develop an electrochemical biosensor, several phosphorylated substrates have been used. Promising results have been achieved with the commercial enzymes and alpha-naphtyl phosphate, p-aminophenol phosphate and catechol monophosphate as enzyme substrates, guaranteeing the viability of the electrochemical approach.

Biosensing Techniques↗

Bipolar II vs. unipolar depression: psychopathologic differentiation by dimensional measures.

BACKGROUND: Clinical presentations of depression in bipolar disorder are varied, inconsistent and often confusing. Most previous studies have focused on bipolar I (BP-I). Given that bipolar II (BP-II) is the more common bipolar phenotype, which is often confused with unipolar (UP), the aim of the current analyses is to delineate the symptomalogic differences between BP II vs. UP MDD in a large national sample. METHODS: The data derived from the French National EPIDEP study (n = 452 DSM-IV major depressives), subdivided into BP-II (n = 196) and UP (n = 256). The BP II group included major depressives with both spontaneous and antidepressant-associated hypomania based on our finding of similarity in rates of familial bipolarity in the two subgroups. At index presentation, depression was assessed by the clinician (using HAM-D and the Rosenthal Atypical Depression Scale) and by the patient (using the Multi-Visual Analog Scale of Bipolarity, MVAS-BP). Principal component analyses (PCA with varimax rotation) were conducted on HAM-D and MVAS-BP in the total population and separately in BP-II and UP. We performed inter-group comparative tests (UP vs. BP-II) on factorial scores derived from PCAs and correlation tests between these factorial scores. RESULTS: The PCA on "HAM-D + Rosenthal scale" showed the presence of nine major factors: F1-2 "weight changes", F3-4 "sleep disturbances", F5 "sadness-guilt", F6 "retardation-fatigue", F7 "somatic", F8 "diurnal variation" and F9 "insight-delusion". The PCA on MVAS-BP revealed the presence of eight principal components: F1 "psychomotor retardation", F2 "central pain", F3 "somatic", F4 "social contact", F5 "worry", F6 "loss of interest", F7 "guilt" and F8 "anger". Despite uniformity in global intensity of depression, significant differences were observed as follows: higher score on "psychomotor retardation" (p = 0.03), "loss of interest" (p = 0.057) and "insomnia" (p = 0.05) in the UP group, and higher score on "hypersomnia" (p = 0.008) in the BP-II group. Correlation analyses between clinician- and self-rating revealed the presence of higher number of significant coefficients in the UP vs. BP-II group (p < or =0.001). LIMITATION: A three-way comparison between BP-I, BP-II and UP may have yielded somewhat different results. CONCLUSION: Our data indicate greater psychomotor retardation, stability and uniformity in the clinical picture of strictly defined UP depression. By contrast, bipolar II depression appeared to be characterized, despite the hypersomnic tendency, by psychomotor activation. This would indicate greater mixed features than those observed in UP. Moreover, in BP-II, there was less agreement between clinician vs. self-rating on the presence of various features of depression. Taken together, these findings explain why BP-II depression is missed by clinicians as a genuine depression.

Antidepressive Agents↗