Parent vs burn-injured child self-report: contributions to a better understanding of anxiety levels.

Anxiety disorders are the most commonly reported form of mental health problem among youth, but they often go undiagnosed and untreated. This study examined the relationship between burn-injured youths' self-reported anxiety levels, as compared with their parent's perceptions of their child's emotional well-being. Burn-injured children were invited to voluntarily complete the Child Version of the 41-item survey, Screen for Child Anxiety Related Disorders, which consists of five anxiety subscales as well as a Total Anxiety Score. Parents were invited to complete the Parent Version. Sixty-three parent-child dyads, with girls (57%) and boys (43%), completed surveys. Mothers (73%) fathers (16%), and other caregivers (11%) participated. Youth mean age was 12.63 years and 60% reported visible burn scars. Matched-pairs t-tests were used to compare parent and child reports. Significantly lower mean scores were found between Screen for Child Anxiety Related Disorders Total Anxiety Score--Parent mean score of 10.52 vs the Youth mean score 21.06 (P<.001), as well as on all subscales including; panic disorder/somatic symptoms (P<.001), generalized anxiety disorder (P=.004), social anxiety disorder (separation anxiety (P<.001), and school avoidance (P<0.001). Results indicate that parents may be severely underestimating the psychological well-being of burn-injured youth. Findings emphasize the importance of a comprehensive approach in assessment for anxiety, involving the collection of feedback from both child and parent. Asking children for input into their psychological well-being is important. This study reinforces the need for a course of ongoing patient and parent education.

Progress in understanding fatigue associated with breast cancer treatment.

Fatigue is one of the most common and distressing symptoms reported by cancer patients. This article reviews research that has examined the extent to which breast cancer patients experience fatigue during and following completion of chemotherapy and radiotherapy. The article also addresses methodological issues in the study of fatigue as well as the current status of efforts to prevent or relieve fatigue associated with breast cancer treatment.

Metallocyclodextrins as building blocks in noncovalent assemblies of photoactive units for the study of photoinduced intercomponent processes.

Cyclodextrin cups have been employed to build supramolecular systems consisting of metal and organic photoactive/redox-active components; the photoinduced communication between redox-active units assembled in water via noncovalent interactions is established. The functionalization of a beta-cyclodextrin with a terpyridine unit, ttp-beta-CD, is achieved by protection of all but one of the hydroxyl groups by methylation and attachment of the ttp unit on the free primary hydroxyl group. The metalloreceptors [(beta-CD-ttp)Ru(ttp)][PF(6)](2), [(beta-CD-ttp)Ru(tpy)][PF(6)](2), and [Ru(beta-CD-ttp)(2)][PF(6)](2) are synthesized and fully characterized. The [(beta-CD-ttp)Ru(ttp)][PF(6)](2) metalloreceptor exhibits luminescence in water, centered at 640 nm, from the (3)MLCT state with a lifetime of 1.9 ns and a quantum yield of Phi = 4.1 x 10(-)(5). Addition of redox-active quinone guests AQS, AQC, and BQ to an aqueous solution of [(beta-CD-ttp)Ru(ttp)](2+) results in quenching of the luminescence up to 40%, 20%, and 25%, respectively. Measurement of the binding strength indicates that, in saturation conditions, 85% for AQS and 77% for AQC are bound. The luminescence quenching is attributed to an intercomponent electron transfer from the appended ruthenium center to the quinone guest inside the cavity. Control experiments demonstrate no bimolecular quenching at these conditions. A photoactive osmium metalloguest, [Os(biptpy)(tpy)][PF(6)], is designed with a biphenyl hydrophobic tail for insertion in the cyclodextrin cavity. The complex is luminescent at room temperature with an emission band maximum at 730 nm and a lifetime of 116 ns. The osmium(III) species are formed for the study of photoinduced electron transfer upon their assembly with the ruthenium cyclodextrin, [(beta-CD-ttp)Ru(ttp)](2+). Time-resolved spectroscopy studies show a short component of 10 ps, attributed to electron transfer from Ru(II) to Os(III) giving an electron transfer rate 9.5 x 10(9) s(-)(1).

3-O-methyl-D-galactose residues in lycophyte primary cell walls.

Acid hydrolysis of cell wall-rich material from young leaves of the lycophyte Selaginella apoda (L.) Spring yielded substantial amounts of 3-O-methyl-D-galactose (1) in addition to the usual major monosaccharides (glucose, galactose, arabinose, xylose and galacturonic acid). The yield of 1 approximately equalled that of galacturonic acid. Compound 1 was identified as 3-O-methylgalactose by its 1H and 13C NMR spectra, and shown to be the D-enantiomer by its susceptibility to D-galactose oxidase. Compound 1 was detected in acid hydrolysates of the alcohol-insoluble residues from young leaves of all lycophytes tested, both homosporous (Lycopodium, Huperzia and Diphasiastrum) and heterosporous (Selaginella). It was not detectable in the charophyte green algae Coleochaete scutata, Chara coralina or Klebsormidium flaccidum, any bryophytes [a hornwort (Anthoceros), four liverworts and three mosses], or any euphyllophytes [a psilopsid (Psilotum), a horsetail (Equisetum), eusporangiate and leptosporangiate ferns, the gymnosperm Gnetum, and diverse angiosperms]. A high content of 1 is thus an autapomorphy of the lycophytes.

The simulated microgravity environment maintains key metabolic functions and promotes aggregation of primary porcine hepatocytes.

The high aspect ratio vessel allows the culture of primary porcine hepatocytes in an environment of low shear stress and simulated microgravity. Primary porcine hepatocytes have been difficult to maintain in culture long term while preserving their metabolic functions. This study was carried out in order to characterise key metabolic functions of cell aggregates formed by primary porcine hepatocytes cultured in a high aspect ratio vessel for a predetermined period of 21 days. 10(8) porcine hepatocytes were loaded into the high aspect ratio vessel and continuously rotated during the experiments. 0.7 ml of the culture medium was sampled on days 1, 2, 4, 7, 10, 14 and 21. 1H nuclear magnetic resonance spectroscopy of the culture medium, using the presaturation technique, assessed the following: glucose metabolism, glutamine synthesis and ketogenesis. There was glucose breakdown anaerobically during the first 10 days as manifested by lactate production and pyruvate and threonine consumption. After day 10 there was significantly smaller lactate production (day 1 vs day 10 P < 0.01), and significantly smaller pyruvate (day 1 vs day 14 P < 0.03) and threonine consumption (day 1 vs day 10 P < 0.002), indicative of an aerobic metabolic pattern. Significantly more glutamate was produced after day 10 (day 1 vs day 10 P < 0.031), and more glutamine was consumed after day 14. There was a steadily diminishing production of acetate which reached a minimum on day 14 (day 2 vs day 14 P < 0.00014). After an initial 10 day period of acclimatisation cell aggregates formed in the high aspect ratio vessel switched from the anaerobic pattern of metabolism to the more efficient aerobic pattern, which was exhibited until the experiments were terminated. The high aspect ratio vessel is suitable for long-term culture of porcine hepatocytes and it is worthwhile carrying out scale-up feasibility studies.

Cultured fish cells metabolize octadecapentaenoic acid (all-cis delta3,6,9,12,15-18:5) to octadecatetraenoic acid (all-cis delta6,9,12,15-18:4) via its 2-trans intermediate (trans delta2, all-cis delta6,9,12,15-18:5).

Octadecapentaenoic acid (all-cis delta3,6,9,12,15-18:5; 18:5n-3) is an unusual fatty acid found in marine dinophytes, haptophytes, and prasinophytes. It is not present at higher trophic levels in the marine food web, but its metabolism by animals ingesting algae is unknown. Here we studied the metabolism of 18:5n-3 in cell lines derived from turbot (Scophthalmus maximus), gilthead sea bream (Sparus aurata), and Atlantic salmon (Salmo salar). Cells were incubated in the presence of approximately 1 microM [U-14C]18:5n-3 methyl ester or [U-14C]18:4n-3 (octadecatetraenoic acid; all-cis delta6,9,12,15-18:4) methyl ester, both derived from the alga Isochrysis galbana grown in H14CO3-, and also with 25 microM unlabeled 18:5n-3 or 18:4n-3. Cells were also incubated with 25 microM trans delta2, all-cis delta6,9,12,15-18:5 (2-trans 18:5n-3) produced by alkaline isomerization of 18:5n-3 chemically synthesized from docosahexaenoic acid (all-cis delta4,7,10,13,16,19-22:6). Radioisotope and mass analyses of total fatty acids extracted from cells incubated with 18:5n-3 were consistent with this fatty acid being rapidly metabolized to 18:4n-3 which was then elongated and further desaturated to eicosatetraenoic acid (all-cis delta8,11,14,17,19-20:4) and eicosapentaenoic acid (all-cis delta5,8,11,14,17-20:5). Similar mass increases of 18:4n-3 and its elongation and further desaturation products occurred in cells incubated with 18:5n-3 or 2-trans 18:5n-3. We conclude that 18:5n-3 is readily converted biochemically to 18:4n-3 via a 2-trans 18:5n-3 intermediate generated by a delta3,delta2-enoyl-CoA-isomerase acting on 18:5n-3. Thus, 2-trans 18:5n-3 is implicated as a common intermediate in the beta-oxidation of both 18:5n-3 and 18:4n-3.

The biosynthetic incorporation of the intact leucine skeleton into sterol by the trypanosomatid Leishmania mexicana.

The amino acid leucine is efficiently used by the trypanosomatid Leishmania mexicana for sterol biosynthesis. The incubation of [2-(13)C]leucine with L. mexicana promastigotes in the presence of ketoconazole gave 14alpha-methylergosta-8,24(24(1))-3beta-ol as the major sterol, which was shown by mass spectrometry to contain up to six atoms of (13)C per molecule. (13)C NMR analysis of the 14alpha-methylergosta-8,24(24(1))-3beta-ol revealed that it was labeled in only six positions: C-2, C-6, C-11, C-12, C-16, and C-23. This established that the leucine skeleton is incorporated intact into the isoprenoid pathway leading to sterol; it is not converted first to acetyl-CoA, as in animals and plants, with utilization of the acetyl-CoA to regenerate 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA). An inhibitor of HMG-CoA synthase (L-659,699) blocked the incorporation of [1-(14)C]acetate into sterol but had no inhibitory effect on [U-(14)C]leucine incorporation. The HMG-CoA reductase inhibitor lovastatin inhibited promastigote growth and [U-(14)C]leucine incorporation into sterol. The addition of unlabeled mevalonic acid (MVA) overcame the lovastatin inhibition of growth and also diluted the incorporation of [1-(14)C]leucine into sterol. These results are compatible with two routes by which the leucine skeleton may enter intact into the isoprenoid pathway. The catabolism of leucine could generate HMG-CoA that is then directly reduced to MVA for incorporation into sterol. Alternatively, a compound produced as an intermediate in leucine breakdown to HMG-CoA (e.g. dimethylcrotonyl-CoA) could be directly reduced to produce an isoprene alcohol followed by phosphorylation to enter the isoprenoid pathway post-MVA.

Pyrogenic toxin superantigen site specificity in toxic shock syndrome and food poisoning in animals.

Staphylococcus aureus and Streptococcus pyogenes express pyrogenic toxin superantigens (PTSAgs) that are associated with toxic shock syndrome (TSS) and staphylococcal food poisoning (SFP). Most PTSAgs cause TSS in deep-tissue infections, whereas only TSS toxin 1 (TSST-1) is associated with menstrual, vaginal TSS. In contrast, SFP has been linked only with staphylococcal enterotoxins (SEs). Because of the differential abilities of PTSAgs to cause systemic or localized symptoms in a site-dependent manner, the present study was undertaken to assess the toxins' abilities to cross mucosal barriers. The activity of three PTSAgs when delivered orally, vaginally, or intravenously to rabbits and orally to monkeys was investigated. TSST-1 induced shock via all three routes in rabbits. Although active when administered intravenously, SEC1 and streptococcal pyrogenic exotoxin A (SPEA) did not cause symptoms when administered orally or vaginally. Only SEC1 induced emesis in the monkey feeding assay. TSST-1, albeit less stable than SEC1 and SPEA to pepsin, induced diarrhea in monkeys. Our results may explain the unique association of TSST-1 with menstrual TSS and why SPEA is only rarely associated with TSS after pharyngitis, despite being highly associated with TSS after subcutaneous infections. Finally, our studies indicate that enterotoxicity in SFP is not the result of superantigenicity.

A linear endothelin-1 analogue: solution structure of ET-1[Aib1,3,11,15, Nle7] by nuclear magnetic resonance spectroscopy and molecular modelling.

Two-dimensional nuclear magnetic resonance techniques and a combination of distance geometry and molecular dynamics calculations were utilised to determine the three dimensional solution structure of an ET-1 analogue, ET-1[Aib1,3,11,15, Nle7], in a methanol-d3/water co-solvent. The modelled structure shows that the peptide folds into a consistent alpha-helical conformation between residues Ser4-His16 while the C-terminus prefers no fixed conformation. Our studies confirm that the disulphide links which are normally associated with the endothelin family of neuropeptides are not important for the formation of a helical conformation in solution. This full length, modified, synthetic linear ET-1 analogue plays a vital role towards designing endothelin receptor agonists. Structure activity relationships are discussed in terms of the conformational features of the calculated structure.

Solution structure of a novel ETB receptor selective agonist ET1-21 [Cys(Acm)1,15, Aib3,11, Leu7] by nuclear magnetic resonance spectroscopy and molecular modelling.

The solution structure of a biologically active modified linear endothelin-1 analogue, ET1-21[Cys(Acm)1,15, Aib3,11, Leu7], has been determined for the first time by two-dimensional nuclear magnetic resonance spectroscopy in a methanol-d3/water solvent mixture. Out of approximately one hundred linear peptide analogues tested by biological assay, this peptide, together with a dozen others, showed significant ETB selective agonist activity. Here we report the solution structure of an ETB selective agonist of a full-length, synthetic linear endothelin analogue. The calculated structures indicate that the peptide adopts an alpha-helical conformation between residues Ser5-His16, whilst both N- and C-termini show no preferred conformation. These results suggest that the disulphide bridges normally associated with endothelin and sarafotoxin peptides may not necessarily be important for either ETB receptor binding activity or the formation of a helical conformation in solution.

Development of ET(B) selective agonists: solution structure of a linear endothelin-1 analogue, ET-1 [Cys(Acm)(1,15), Ala3, Leu7, dAsp8, Aib11].

The solution structure of a synthetic ET(B) selective agonist, ET-1[Cys(Acm)(1,15), Ala3, Leu7, dAsp8, Aib11] has been solved by 1H NMR and molecular modelling studies. Such solution structures of linear modified peptides in aqueous methanol are being used in an ongoing program of research designed to assist in an understanding of the basic structural requirements for the biological activity of vasoconstrictors. The resulting structure of this peptide is characterised by an alpha-helical conformation between residues Leu6-His16 and by N- and C-termini which assume no defined conformation. A knowledge of the solution structures of this and related peptides, which are ET(B) selective agonists, are proving to be important in the understanding of how they interact with the ET(B) receptor.

Solution conformation of an ET(B) selective agonist, ET-1[Cys(Acm)1,15,Ala3,Leu7,Aib11], in CD3OH/H2O by 1H NMR and molecular modelling.

To understand the basic structural requirements for the biological activity of endothelin peptides, the solution structure of an ETB selective agonist, ET-1[Cys-(Acm)1,15, Ala3,Leu7,Aib11, was investigated by 1H NMR spectroscopy and molecular modelling. The structure is characterised by an alpha-helical conformation between residues Ser5-His16 but is undefined at both the N and C termini. To date, neither the solution structures of linear modified peptides nor the effects of a methanol/water solvent system have been examined for endothelin or endothelin-like peptides. This structure plays an important role towards the design of endothelin receptor selective agonists and antagonists.

Pulcherosine, an oxidatively coupled trimer of tyrosine in plant cell walls: its role in cross-link formation.

An oxidatively coupled trimer of tyrosine has been isolated from hydrolysates of primary cell walls of a tomato cell culture. UV-absorption, fluorescence and 1H NMR spectra showed that the trimer was pulcherosine, composed of isodityrosine and tyrosine oxidatively coupled via a biphenyl linkage such that the aromatic core is 2,2'-dihydroxy-3-phenoxybiphenyl. Pulcherosine could act as an intermediate in the conversion of isodityrosine to the tetramer, di-isodityrosine. Steric considerations show that the three tyrosine units of pulcherosine could not be near-neighbour residues within a single polypeptide chain. Pulcherosine therefore forms inter-polypeptide cross-links and/or wide intra-polypeptide loops.

N alpha- and N epsilon-D-galacturonoyl-L-lysine amides: properties and possible occurrence in plant cell walls.

Three representatives of a novel class of amide (isopeptide) glycoconjugates have been synthesised: N alpha-D-galacturonoyl-L-lysine and N epsilon-D-galacturonoyl-L-lysine and N epsilon-D-polygalacturonoyl-L-lysine. Galacturonoyl-lysine amide bonds were labile in 2 M trifluoroacetic acid at 120 degrees and in alkali, but relatively stable in cold acid. The amide bonds were resistant to digestion by Driselase, Pronase and trypsin. The polysaccharide backbone of N epsilon-D-polygalacturonoyl-L-lysine was hydrolysed by Driselase to yield two major ninhydrin-positive compounds which were shown by 1H and 13C NMR spectroscopy to be tri- and tetra-alpha-(1-->4)-D-galacturonoyl-L-lysines. To investigate the possible natural occurrence of N-galacturonoyl isopeptide bonds, we fed cell-suspension cultures of spinach and tomato with D-[6-14C]glucuronic acid, which radio-labels pectic polysaccharides. The radioactive cell walls were digested with, sequentially, Driselase, mild acid, and proteinases. On electrophoresis at pH 2.0, several of the radioactive digestion-products were cathodic. Some of the cathodic products yielded [14C]galacturonic acid upon complete acid hydrolysis. The existence of these products is compatible with the presence of novel N-galacturonoyl isopeptide bonds, which could serve as cross-links in plant cell walls.

Bias effects in three common self-report pain assessment measures.

OBJECTIVE: Past research has shown response biases to influence the accuracy of results from self-report measures. In pain assessment, where a percentage of patients have financial and other reasons to minimize or exaggerate psychological disturbance, it becomes especially important to identify the influence of response bias in self-report of adjustment. This study investigated the susceptibility of three commonly used self-report pain assessment measures to response bias. DESIGN: This study used a within-subjects (asymptomatic subjects) design with two experimental conditions and nonequivalent control group (chronic pain patients). SUBJECTS: Experimental group: 40 students enrolled in an occupational therapy program at a major southeastern United States university. CONTROL GROUP: 200 subjects referred to a multidisciplinary pain clinic at a major teaching hospital. MEASURES: Coping Strategies Questionnaire, Multidimensional Pain Inventory, and Pain Beliefs and Perceptions Inventory. RESULTS: With few exceptions, asymptomatic subjects scored significantly differently on these measures while portraying themselves as either coping well or coping poorly. In addition, when using the "coping poorly" response set, asymptomatic subjects reproduced scores similar to those of symptomatic chronic pain patients. CONCLUSION: The susceptibility to manipulation appeared constant across the three measures, a finding that highlighted the difficulties clinicians and researchers encounter in accurate interpretation of results from these measures in the absence of validity indicators. This study also emphasizes the ease with which subjects with sufficient motivation can present themselves in an untruthful and manipulative manner and can generate scores that are, on their own, difficult to distinguish from those of a group of typical chronic pain patients.

The Coping Strategies Questionnaire: a large sample, item level factor analysis.

OBJECTIVE: The Coping Strategies Questionnaire (CSQ), a measure of coping in chronic pain patients, was subjected to item-level exploratory factor analysis. SUBJECTS: A sample of 965 chronic pain patients were used in the analysis. RESULTS: Principal components analysis using a varimax rotation procedure identified nine factors that accounted for 54.5% of the variance. Of these nine factors, the first five represent subscales of the original CSQ subscales. The catastrophizing subscale replicated with significant loadings for all six original items, and ignoring sensations replicated with five of six items. Factors representing reinterpreting pain sensations, coping self-statements, and diverting attention subscales also appeared. The items from the praying and hoping subscale split into separate praying and hoping factors (factors 6 and 8). When reliability coefficients were calculated, factors 7 through 9 had unacceptably low internal consistency and thus were not considered stable factors. Correlations between factors 1 through 6 and other measures of psychological and physical functioning were calculated in the construct validation portion of this study. Previously found relationships were replicated in that the correlations between CSQ factor scores and measures of pain, depression, and disability were in the same direction in this data set as those previously reported.

Solution structure determination of endothelin-1 in methanol/water by NMR and molecular modelling methods.

To understand the structural requirements for the biological activity of endothelin peptides and to develop receptor selective endothelin analogues further, the solution structure of the bicyclic 21 amino acid residue vasoactive peptide, endothelin-1, has been determined in methanol-d3/water using high-resolution 1H-NMR spectroscopy. To our knowledge, this solvent system has not previously been used in NMR studies of endothelin and/or endothelin-like peptides. Two-dimensional DQFCOSY, TOCSY and NOESY spectra were acquired along with a series of one-dimensional spectra. A total of 219 distance constraints and 5 angle constraints were derived from the NMR data. These were incorporated into structure calculations using distance geometry (DIANA) followed by simulated annealing and molecular dynamics. The resulting structures are characterized by an alpha-helical conformation, Lys9-His16, and residues Ser5-Asp8 form a type I beta-turn. The N-terminal region, which was not extensively constrained by NMR data, showed no preferred conformation. The C-terminal tail showed less extensive conformational averaging but no descriptive conformation could be observed. The results obtained in this study are in good agreement with previous proposals.

Di-isodityrosine, a novel tetrametric derivative of tyrosine in plant cell wall proteins: a new potential cross-link.

A novel amino acid, di-isodityrosine, has been isolated from hydrolysates of cell walls of tomato cell culture. Analysis by UV spectrometry, partial derivatization with 2,4-dinitrofluorobenzene and mass and NMR spectrometry show that the compound is composed to two molecules of isodityrosine, joined by a biphenyl linkage. The possible reactions involved in the formation of this molecule in vivo are discussed, as is the possibility that it could form an interpolypeptide linkage between cell wall proteins such as extensin, and hence aid in the insolubilization of the protein in the wall.

Sulphated compounds attenuate beta-amyloid toxicity by inhibiting its association with cells.

Agents that interfere with the toxic effects of beta-amyloid protein may be therapeutically useful against Alzheimer's disease. We reported recently that several sulphated glycosaminoglycans and sulphonated dyes attenuate the toxic effects of beta-amyloid fragments beta 25-35 and beta 1-40 in two clonal cell lines. We now demonstrate that this protective effect is due to interference with beta-amyloid cell association rather than effects on beta-amyloid structure. Using an enzyme-linked immunoabsorbance assay to detect cell-associated beta 1-40, we found in a range of compounds a strong correlation between inhibition of HeLa cell association of beta 1-40 and attenuation of cellular toxicity as measured by inhibition of 3-[4,5-dimethylthia-zol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) reduction. In contrast, effects on peptide structure, as measured by Congo red binding, were generally inconsistent with the attenuating effects of the compounds on cellular toxicity. These results suggest that by binding beta-amyloid these agents prevent its interaction with cells.

Sulfonated dyes attenuate the toxic effects of beta-amyloid in a structure-specific fashion.

We recently reported that several sulfate-containing glycosaminoglycans, a class of compounds associated with the beta-amyloid plaques of Alzheimer's disease, attenuate the toxic effects of beta-amyloid fragments beta 25-35 and beta 1-40. The amyloid-binding sulfonated dye Congo Red was shown to have a similar effect. Using two clonal cell lines, we now demonstrate that several sulfonated dyes attenuate beta-amyloid toxicity and that the protective effect appears specific for compounds whose sulfonate groups can interact with the beta-pleated structure of aggregated amyloid. These results suggest that by binding beta-amyloid these compounds may prevent toxic interactions of the peptide with cells.