Activation of beta(2)-adrenergic receptors hastens relaxation and mediates phosphorylation of phospholamban, troponin I, and C-protein in ventricular myocardium from patients with terminal heart failure

Background-Catecholamines hasten cardiac relaxation through beta-adrenergic receptors, presumably by phosphorylation of several proteins, but it is unknown which receptor subtypes are involved in human ventricle. We assessed the role of beta(1)- and beta(2)-adrenergic receptors in phosphorylating proteins implicated in ventricular relaxation. Methods and Results-Right ventricular trabeculae, obtained from freshly explanted hearts of patients with dilated cardiomyopathy (n=5) or ischemic cardiomyopathy (n=5), were paced at 60 bpm. After measurement of the contractile and relaxant effects of epinephrine (10 mu mol/L) or zinterol (10 mu mol/L), mediated through beta(2)-adrenergic receptors, and of norepinephrine (10 mu mol/L), mediated through beta(1)-adrenergic receptors, tissues were freeze clamped. We assessed phosphorylation of phospholamban, troponin I, and C-protein, as well as specific phosphorylation of phospholamban at serine 16 and threonine 17, Data did not differ between the 2 disease groups and were therefore pooled. Epinephrine, zinterol, and norepinephrine increased contractile force to approximately the same extent, hastened the onset of relaxation by 15+/-3%, 5+/-2%, and 20+/-3%, respectively, and reduced the time to half-relaxation by 26+/-3%, 21+/-3%, and 37+/-3%. These effects of epinephrine, zinterol, and norepinephrine were associated with phosphorylation (pmol phosphate/mg protein) of phospholamban 14+/-3, 12+/-4, and 12+/-3, troponin I 40+/-7, 33+/-7, and 31+/-6; and C-protein 7.2+/-1.9, 9.3 +/- 1.4, and 7.5 +/- 2.0. Phosphorylation of phospholamban occurred at both Ser16 and Thr17 residues through both beta(1)- and beta(2)-adrenergic receptors. Conclusions-Norepinephrine and epinephrine hasten human ventricular relaxation and promote phosphorylation of implicated proteins through both beta(1)- and beta(2)-adrenergic receptors, thereby potentially improving diastolic function.

Structures of free and complexed forms of Escherichia coli xanthine-guanine phosphoribosyltransferase

Structures of free, substrate-bound and product-bound forms of Escherichia coli xanthine-guanine phosphoribosyltransferase (XGPRT) have been determined by X-ray crystallography. These are compared with the previously determined structure of magnesium and sulphate-bound XPRT. The structure of free XGPRT at 2.25 Angstrom resolution confirms the flexibility of residues in and around a mobile loop identified in other PRTases and shows that the cis-peptide conformation of Arg37 at the active site is maintained in the absence of bound ligands. The structures of XGPRT complexed with the purine base substrates guanine or xanthine in combination with cPRib-PP, an analog of the second substrate PRib-PP, have been solved to 2.0 Angstrom resolution. In these two structures the disordered phosphate-binding loop of uncomplexed XGPRT becomes ordered through interactions with the 5'-phosphate group of cPRib-PP. The cyclopentane ring of cPRib-PP has the C3 exo pucker conformation, stabilised by the cPRib-PP-bound Mg2+. The purine base specificity of XGPRT appears to be due to water-mediated interactions between the 2-exocyclic groups of guanine or xanthine and side-chains of Glu136 and Asp140, as well as the main-chain oxygen atom of Ile135. Asp92, together with Lys115, could help stabilise the N7-protonated tautomer of the incoming base and could act as a general base to remove the proton from N7 .when the nucleotide product is formed. The 2.6 Angstrom resolution structure of XGPRT complexed with product GMP is similar to the substrate-bound complexes. However, the ribose ring of GMP is rotated by similar to 24 degrees compared with the equivalent ring in cPRib-PP. This rotation results in the loss of all interactions between the ribosyl group and the enzyme in the product complex. (C) 1998 Academic Press.

Anxious children and their parents: What do they expect?

Assigned 73 children, ages 7 to 14, to 1 of 3 groups (anxious, clinical control, and nonclinical control) according to their diagnostic status. Within the anxious group, children were assigned to 1 of 2 further groups on the basis of self-reported parental anxiety-either the child anxiety only group or the child + parent anxiety group. All children completed an experimental task (giving a brief talk in front of a video camera), which was the focus for a series of structured family discussions between the child and his or her parents. The aims of the study were to measure and compare across groups (a) the evaluations of children and their parents regarding the child's predicted anxiety and skill level and (b) the effect of the family discussion on children's expectations. Results indicated that, prior to the family discussion, anxious children's expectations of their future performance did not differ from those of control children. Similarly, there were no differences in children's expectations between the child anxiety group and the child + parent anxiety group. Second, compared to mothers in the child anxiety group, mothers in the child + parent anxiety group expected that their children would be more anxious and more likely to choose an avoidant problem solution (but not less skilled). Finally, the family discussion was found to produce no changes in anxious children's expectations of their future performance. The implications of these findings are discussed.

Timing of rotator cuff activation during shoulder external rotation in throwers with and without symptoms of pain

Study Design: Fine-wire EMG rotator cuff onset time analysis in 2 matched groups of throwers with and without pain. Objective: To identify if there is a difference in the activation patterns of the rotator cuff muscles during a rapid shoulder external rotation task between throwers with and without pain. Background: The coordinated action of the rotator cuff is recognized as essential for glenohumeral joint control in the throwing athlete. Identification of abnormalities occurring in muscle activation patterns for injured athletes is relevant when prescribing rehabilitative exercises. Methods and Measures: Twelve throwers with shoulder pain were compared to a matched group of 11 asymptomatic throwers. Participants were matched for age, height, body mass, and habitual activity. Fine-wire EMG electrodes were inserted into the subscapularis, supraspinatus, and infraspinatus. EMG activity was measured during a reaction time task of rapid shoulder external rotation in a seated position. The timing of onset of EMG activity was analyzed in relation to visualization of a light (reaction time) and to the onset of infraspinatus activity (relative latency). Results: In the group with shoulder pain, the onset of subscapularis activity was found to be significantly delayed (reaction time, P = .0018; relative latency, P = .0005) from the onset of infraspinatus activity when compared to the control group. Conclusions: The presence of shoulder pain in these athletes was associated with a difference in the onset of subscapularis EMG activity during a rapid shoulder external rotation movement. This was an initial step in the understanding of the joint protection mechanisms of the glenohumeral joint and the problems that occur in throwers. This information may assist in providing future guidelines for more effective rehabilitation and prevention strategies for this condition.

Morphometric and cladistic analyses of the phylogeny of Macropodinium (Ciliophora : Litostomatea : Macropodiniidae)

Phylogenetic studies of the genus Macropodinium were conducted using two methods; phenetics and cladistics. The phenetic study of morphometrics suggested that the genus could be divided into 3 groups attributable mostly to cell size and shape. The cladistic study also split the genus into 3 groups related to cell size but groups were further distinguished by patterns of ornamentation. Reconciliation of both approaches revealed considerable congruence, however, it also suggested the existence of convergences in the phenetic study and a lack of resolution in the cladistic study. The morphological diversity of Macropodinium is probably due to evolutionary trends such as increasing body size, allometry and polymerisation of structures. None of these trends, however, was uniformly directional and differential effects were observed in different regions of the phylogenetic tree. Comparison of the phylogeny of Macropodinium to a consensus phylogeny of the macropodids revealed limited incongruence between the 2 trees. The ciliate groups could be related to 2 host groups; the wallaby genera and the kangaroo and wallaroo subgenera. The association with these host groups may be the result of phyletic codescent, ecological resource tracking or a combination of both. Further studies of both host and ciliate phylogeny are necessary to resolve these effects.

Milling quality and protein properties of autumn-sown Chinese wheats evaluated through multi-location trials

Eight milling quality and protein properties of autumn-sown Chinese wheats were investigated using 59 cultivars and advanced lines grown in 14 locations in China from 1995 to 1998. Wide ranges of variability for all traits were observed across genotypes and locations. Genotype, location, year, and their interactions all significantly influenced most of the quality parameters. Kernel hardness, Zeleny sedimentation value, and mixograph development time were predominantly influenced by the effects of genotype. Genotype, location and genotype x location interaction were all important sources of variation for thousand kernel weight, test weight, protein content, and falling number, whereas genotype x location interaction had the largest effect on flour yield. Most of the genotypes were characterized by weak gluten strength with Zeleny sedimentation values less than 40 ml and mixograph development time shorter than 3 min. Eight groups of genotypes were recognized based on the average quality performance, grain hardness and gluten strength were the two parameters that determined the grouping, with contributions from protein content. Genotypes such as Zhongyou 16 and Annong 8903 displayed good milling quality, high grain hardness, protein content and strong gluten strength with high sedimentation value and long mixograph development time. Genotypes such as Lumai 15 and Yumai 18 were characterized by low grain hardness, protein content and weak gluten strength. Genotypes such as Yannong 15 and Chuanmai 24 were characterized by strong gluten strength with high sedimentation value and long mixograph development time, but low grain hardness and protein content lower than 12.3%. Genotypes such as Jingdong 6 and Xi'an 8 had weak gluten strength, but with high grain hardness and protein content higher than 12.2%. Five groups of locations were identified, and protein content and gluten strength were the two parameters that determined the grouping. Beijing, Shijiazhuang, Nanyang, Zhumadian and Nanjing produced wheats with medium to strong gluten strength and medium protein content, although there was still a large variation for most of the traits investigated between the locations. Wheat produced in Yantai was characterized by strong gluten strength, but with low protein content. Jinan, Anyang and Linfen locations produced wheats with medium to weak gluten strength and medium to high protein content. Wheats produced in Yangling, Zhenzhou, and Chengdu were characterized by weak gluten strength with medium to low protein content, whereas wheats produced in Xuzhou and Wuhan were characterized by weak gluten strength with low protein content. Industrial grain quality could be substantially improved through integrating knowledge of geographic genotype distribution with key location variables that affected end-use quality.

Body composition in children in remission from acute lymphoblastic leukemia

Background: Changes in body composition are commonly reported in pediatric survivors of acute lymphoblastic leukemia (ALL). However, the effect of ALL and of its treatment on body composition in children in remission from ALL has not been fully examined with the use of a reference method. Objectives: We aimed to determine the body composition and composition of fat-free mass (FFM) in children in remission from ALL. We also aimed to compare the effects that prednisolone and dexamethasone had on the body composition of an ALL survivor population. Design: This cross-sectional study measured height, weight, body volume, total body water, and bone mineral content in 24 children in remission from ALL and 24 age-matched, healthy control subjects. Body composition and FFM composition were evaluated by using the 4-component model. Results: The mean body mass index and fat mass index were significantly (P = 0.05 for both) higher in the ALL survivors than in age-matched control subjects. The composition of the FFM in the 2 treatment groups was not observed to differ significantly. Examination of the composition of FFM made it evident that children in remission from ALL had both significantly greater hydration (P = 0.001) and lower density (P = 0.0001) of FFM than did the control children. Conclusions: Children in remission from ALL may develop excess body fat. To measure body composition accurately in an ALL population, the high hydration and low density of FFM in this population should be taken into consideration.

Ten Australian elementary teachers' discourse and reported pedagogical practices during cooperative learning

This article describes the types of discourse 10 Australian grade 4-6 teachers used after they had been trained to embed cooperative learning in their curriculum and to use communication skills to promote students' thinking and to scaffold their learning. One audiotaped classroom social science lesson involving cooperative learning was analyzed for each teacher. We provide vignettes from 2 teachers as they worked with groups and from 2 student groups. The data from the audiotapes showed that the teachers used a range of mediated-learning behaviors in their interactions with the children that included challenging their perspectives, asking more cognitive and metacognitive questions, and scaffolding their learning. In turn, in their interactions with each other, the children modelled many of the types of discourse they heard their teachers use. Follow-up interviews with the teachers revealed that they believed it was important to set expectations for children's group behaviors, teach the social skills students needed to deal with disagreement in groups, and establish group structures so children understood what was required both from each other and the task. The teachers reported that mixed ability and gender groups worked best and that groups should be no larger than 5 students. All teachers' programs were based on a child-centered philosophy that recognized the importance of constructivist approaches to learning and the key role interaction plays in promoting social reasoning and learning.

AFLP analysis of genetic diversity within Saccharum officinarum and comparison with sugarcane cultivars

Molecular diversity among 421 clones of cultivated sugarcane and wild relatives was analysed using AFLP markers. Of these clones, 270 were Saccharum officinarum and 151 were either cultivars produced by the Australian breeding program or important parents used in the breeding program. The S. of. cinarum clones were obtained from a collection that contained clones from all the major regions where S. of. cinarum is grown. Five AFLP primer combinations generated 657 markers ofwhich 614 were polymorphic. All clones contained a large number of markers; a result of the polyploid nature and heterozygosity of the genome. S. of. cinarum clones from New Guinea displayed greater diversity than S. of. cinarum clones from other regions. This is in agreement with the hypothesis that New Guinea is the centre of origin of this species. The S. of. cinarum clones from Hawaii and Fiji formed a separate group and may correspond to clones that have been introgressed with other members of the ` Saccharum complex'. Greater diversity was found in the cultivars than in the S. of. cinarum clones due to the introgression of S. spontaneum chromatin. These cultivars clustered as expected based on pedigree. The major contribution of clones QN66- 2008 and Nco310 to Australian sugarcane cultivars divided the cultivars into 2 main groups. Although only a fewS. of. cinarum clones are known to have been used in the breeding of current cultivars, about 90% of markers present in the S. of. cinarum clone collection ( 2n= 80) were also present in the cultivar collection. This suggests that most of the observed genetic diversity in S. of. cinarum has been captured in Australian sugarcane germplasm.

Low back pain patients demonstrate increased hip extensor muscle activity during standardized submaximal rotation efforts

Study Design. A comparative study of trunk and hip extensor muscle recruitment patterns in 2 subject groups. Objective. To examine for changes in recruitment of the hip and back extensor muscles during low level isometric trunk rotation efforts in chronic low back pain (CLBP) subjects by comparison with matched asymptomatic control subjects. Summary of Background Data. Anatomic and biomechanical models have provided evidence that muscles attaching to the thoracolumbar fascia (TLF) are important for providing stabilization to the lumbopelvic region during trunk rotation. This has guided rehabilitation programs. The muscles that link diagonally to the posterior layer of the TLF have not previously been examined individually and compared during low-level trunk rotation efforts in CLBP patients and matched controls. Methods. Thirty CLBP patients and 30 matched controls were assessed using surface electromyography (EMG) as they performed low-level isometric rotation efforts while standing upright. Muscles studied included latissimus dorsi, erector spinae, upper and lower gluteus maximus, and biceps femoris. Subjects performed the rotation exertion with various levels of external trunk support, related to different functional tasks. Results. EMG results demonstrated that subjects with CLBP had significantly higher levels of recruitment for the lower and upper gluteus maximus (P < 0.05), hamstrings (P < 0.05), and erector spinae muscles (P < 0.05) during rotation to the left compared with the control subjects. Conclusion. This study provided evidence of increased muscle recruitment in CLBP patients when performing a standardized trunk rotation task. These results may have implications for the design of therapeutic exercise programs for CLBP patients.

Macrocyclic systems containing 2,6,9-trioxabicyclo[3.3.1]-nona-3,7-dienes as chiral spacer groups: Synthesis, stereochemical features and preliminary complexation properties

Novel 2:2-macrocycles bearing bridged concave 2,6,9-trioxabicyclo[3.3.1]nona-3,7-dienes as chiral spacer units were obtained by cyclocondensation reaction of the chiral bisacid chloride and the corresponding diols, while use of methylene diamines instead of diols afforded 1:1 macrocycles only. Applying the same, but now template-assisted, experimental procedure to the reaction of the bisacid chloride with triethylene glycol brought about a significant increase in yield as well as a suitable simplification of the work-up during preparation and separation of the corresponding 1:1 as well as 2:2 macrocycles, when compared to results reported previously. HPLC separation on chiral columns revealed the presence of diastereoisomers [RR(S,S)- and RS-(meso)-forms] for all 2:2 macrocycles, which was further evidenced by the CD spectrum of one of those species as an example. Preliminary ESI-MS experiments indicated strong complexation abilities of the sulphur-containing ligand towards Ag(I), Cu(II) and Au(III) ions.

Arachidonic acid release from mammalian cells transfected with human groups IIA and X secreted phospholipase A(2) occurs predominantly during the secretory process and with the involvement of cytosolic phospholipase A(2)-alpha

Stable expression of human groups IIA and X secreted phospholipases A(2) (hGIIA and hGX) in CHO-K1 and HEK293 cells leads to serum- and interleukin-1beta-promoted arachidonate release. Using mutant CHO-K1 cell lines, it is shown that this arachidonate release does not require heparan sulfate proteoglycan- or glycosylphosphatidylinositol-anchored proteins. It is shown that the potent secreted phospholipase A(2) inhibitor Me-Indoxam is cell-impermeable. By use of Me-Indoxam and the cell-impermeable, secreted phospholipase A(2) trapping agent heparin, it is shown that hGIIA liberates free arachidonate prior to secretion from the cell. With hGX-transfected CHO-K1 cells, arachidonate release occurs before and after enzyme secretion, whereas all of the arachidonate release from HEK293 cells occurs prior to enzyme secretion. Immunocytochemical studies by confocal laser and electron microscopies show localization of hGIIA to the cell surface and Golgi compartment. Additional results show that the interleukin-1beta-dependent release of arachidonate is promoted by secreted phospholipase A(2) expression and is completely dependent on cytosolic (group IVA) phospholipase A(2). These results along with additional data resolve the paradox that efficient arachidonic acid release occurs with hGIIA-transfected cells, and yet exogenously added hGIIA is poorly able to liberate arachidonic acid from mammalian cells.