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A training and coaching team embarked on a training and technical assistance program evaluation project to examine the development of a tool used to determine the impact of training on staff and families serving youth with behavioral health challenges and their families. The training evaluation project centered on the development of a tool to examine the impact of training on trainee attitudes. The current article provides an overview of the development of the Nurtured Heart Approach ® (NHA) Questionnaire to help support training and technical assistance implementation efforts by nursing and other trainers interested in best practice approaches to empower youth with behavioral health challenges. The questionnaire was found to be internally consistent and to have relatively stable, empirically derived factors consistent with the NHA model, but differing somewhat from the theoretically derived “Stands,” or areas of emphasis. The questionnaire is sensitive to changes in attitude resulting from the NHA training. Therefore, it would a viable, practical instrument to test staff acquisition of NHA–related attitudes. [ Journal of Psychosocial Nursing and Mental Health Services, 60 (2), 15–19.]

A training and coaching team embarked on a training and technical assistance program evaluation project to examine the development of a tool used to determine the impact of training on staff and families serving youth with behavioral health challenges and their families. The training evaluation project centered on the development of a tool to examine the impact of training on trainee attitudes. The current article provides an overview of the development of the Nurtured Heart Approach

Thermoplasma acidophilum is a thermoacidophilic archaeon that grows optimally at pH 2 and 59°C. This extremophile is remarkable by the absence of a cell wall or an S-layer. Treating the cells with Triton X-100 at pH 3 allowed the extraction of all of the cell surface glycoproteins while keeping cells intact. The extracted glycoproteins were partially purified by cation-exchange chromatography, and we identified five glycoproteins by N-terminal sequencing and mass spectrometry of in-gel tryptic digests. These glycoproteins are positive for periodic acid-Schiff staining, have a high content of Asn including a large number in the Asn-X-Ser/Thr sequon and have apparent masses that are 34-48 larger than the masses deduced from their amino acid sequences. The pooled glycoproteins were digested with proteinase K and the purified glycopeptides were analyzed by NMR. Structural determination showed that the carbohydrate part was represented by two structures in nearly equal amounts, differing by the presence of one terminal mannose residue. The larger glycan chain consists of eight residues: six hexoses, one heptose and one sugar with an unusual residue mass of 226 Da which was identified as 6-deoxy-6-C-sulfo-d-galactose (6-C-sulfo-d-fucose). Mass spectrometry analyses of the peptides obtained by trypsin and chymotrypsin digestion confirmed the principal structures to be those determined by NMR and identified 14 glycopeptides derived from the main glycoprotein, Ta0280, all containing the Asn-X-Ser/Thr sequons. Thermoplasma acidophilum appears to have a "general" protein N-glycosylation system that targets a number of cell surface proteins. © 2012 The Author.

Xenopus Aurora-A (also known as Eg2) is a member of the Aurora family of mitotic serine/threonine kinases. In Xenopus oocytes, Aurora-A phosphorylates and activates a cytoplasmic mRNA polyadenylation factor (CPEB) and therefore plays a pivotal role in MOS translation. However, hyperphosphorylation and activation of Aurora-A appear to be dependent on maturation-promoting factor (MPF) activation. To resolve this apparent paradox, we generated a constitutively activated Aurora-A by engineering a myristylation signal at its N terminus. Injection of Myr-Aurora-A mRNA induced germinal vesicle breakdown (GVBD) with the concomitant activation of MOS, mitogen-activated protein kinase, and MPF. Myr-Aurora-A-injected oocytes, however, appeared to arrest in meiosis I with high MPF activity and highly condensed, metaphase-like chromosomes but no organized microtubule spindles. No degradation of CPEB or cyclin B2 was observed following GVBD in Myr-Aurora-A-injected oocytes. In the presence of progesterone, the endogenous Aurora-A became hyperphosphorylated and activated at the time of MPF activation. Following GVBD, Aurora-A was gradually dephosphorylated and inactivated before it was hyperphosphorylated and activated again. This biphasic pattern of Aurora-A activation mirrored that of MPF activation and hence may explain meiosis I arrest by the constitutively activated Myr-Aurora-A.

Several recent studies have demonstrated that insulin-like growth factor (IGF)-1-induced mitogen-activated protein kinase (MAP kinase) activation is abolished by pertussis toxin, suggesting that trimeric G proteins of the G(i) class are novel cellular targets of the IGF-1 signaling pathway. We report here that the intracellular domain of the Xenopus IGF-1 receptor is capable of binding to the Xenopus homolog of mammalian GIPC, a PDZ domain-containing protein previously identified as a binding partner of G(i)-specific GAP (RGS-GAIP). Binding of xGIPC to xIGF-1 receptor is independent of the kinase activity of the receptor and appears to require the PDZ domain of xGIPC. Injection of two C-terminal truncation mutants that retained the PDZ domain blocked IGF-1-induced Xenopus MAP kinase activation and oocyte maturation. While full-length xGIPC injection did not significantly alter insulin response, it greatly enhanced human RGS-GAIP in stimulating the insulin response in frog oocytes. This represents the first demonstration that GIPC x RGS-GAIP complex acts positively in IGF-1 receptor signal transduction.

We have previously identified an endothelial cell membrane protein of Mr 45 kDa that binds plasminogen in a kringle-dependent, specific and reversible manner (Dudani et. al. (1991) Mol. Cell. Biochem. 108: 133–139). In this study, we have developed and optimized a protocol for the isolation of the 45 kDa plasminogen receptor from venous endothelial cells using a four step procedure consisting of lysis and detergent extraction followed by ligand affinity chromatography and preparative polyacrylamide gel electrophoresis. Control experiments were carried out using BSA-Sepharose instead of plasminogen-Sepharose as the affinity matrix. No plasminogen binding proteins were recovered from the former columns. However, a 45 kDa protein was recovered from lysine eluates of plasminogen-Sepharose. This material was then purified to homogeneity using preperative electrophoresis. Analyses of proteins at various steps in the purification by SDS-PAGE showed enrichment of a band of 45 kDa which superimposed with the observed binding activity of plasminogen in ligand blots. The above binding could be inhibited by excess lysine. The 45 kDa protein could be distinguished from alpha-enolase which also binds plasminogen by (i) significant differences in the profile of retention times of CNBr-degradation fragments on reversed phase HPLC; and (ii) partial peptide sequencing of one of the CNBr-degradation fragments of the 45 kDa protein. Moreover, the derived sequence did not show any significant homology to any protein in the Swiss Prot (release 20) database. We thus propose that the 45 kDa protein represents a novel plasminogen receptor on human venous endothelial cells.

Synthetic peroxovanadium compounds are a new class of potent inhibitors of protein phosphotyrosine phosphatases. These compounds exhibit insulin-like activity bothin vitroand in experimental animals. However, the molecular mechanism by which these compounds exert their biological effect is not well defined. We demonstrate here that several of these compounds induceXenopusoocyte maturationin vitro,as indicated by germinal vesicle breakdown. Using one of these compounds for further studies, we show that the induction is dose-dependent and is accompanied by activation of maturation promoting factor as well as activation ofXenopusMAP kinase. Like insulin, bpV(pic) causes an acute accumulation of PI(3,4,5)P3(phosphotidylinositol-3,4,5-trisphosphate), a product of PI 3-kinase. More importantly, bpV(pic)-induced oocyte maturation was abolished by microinjection of a neutralizing monoclonal anti-insulin receptor antibody (17A3) into oocytes or preincubation of oocytes with a PI 3-kinase inhibitor (wortmannin). These results suggest that bpV(pic) acts upstream of theXenopusIGF-1 receptor in the induction of meiotic maturation, presumably by neutralizing an inhibitory protein tyrosine phosphatase(s) that may regulate the receptor. Finally, using an oocyte–follicle cell complex that responded to human chorionic gonadotropin (hCG) to undergo GVBD, we showed that injection of 17A3 anti-insulin receptor antibody into oocytes did not affect hCG-induced oocyte maturation.