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The roles played by the D2 and D3 domains of recombinant human thrombomodulin in its function.

Thrombomodulin (TM) is composed of five domains. We investigated the roles of the sixth epidermal growth factor (EGF)-like structure (E6) in the second domain (D2) and of an O-glycosylation site rich domain (D3) in the function of TM in more detail using deletion analysis. Two soluble mutants of TM, TMD123 and TMD12, and three deletion mutants lacking respectively 6, 16, and 38 C-terminal residues of the E6 portion, TMD12 delta 6, TMD12 delta 16, and TMD12 delta 38, were expressed in COS cells and purified. The results of protein C-activating cofactor assay showed that TMD12 delta 6, TMD12 delta 16, and TMD12 delta 38, which lack the C-terminal region, had remarkably weak cofactor activities in comparison with TMD123 (9.1, 1.4, and 1.1% of TMD123 activity, respectively). Similar findings were obtained for anticoagulant activity. These findings indicate that the last loop structure in E6 is required for full activity of recombinant human TM. We also determined in vivo stabilities of TMD12, TMD123, and TMD12 delta 6 in a pharmacokinetic study in rats. TMD12 and TMD12 delta 6, which lack the D3 domain, exhibited increased clearance (about twice that of D123). This finding suggested that the D3 domain of TM plays an important role in stabilizing TM in vivo.

Anticoagulants↗

Etiology of spondylolisthesis. Assessment of the role played by lumbar facet joint morphology.

This study examined the role of facet joint morphology in the etiology of both degenerative spondylolisthesis and isthmic spondylolysis. To this end, the axial facet joint morphology of the lower lumbar spine in a normal population and in populations of patients with spinal stenosis or degenerative spondylolisthesis at L4-5 and in patients with isthmic spondylolysis at the L5 level were characterized. Computed tomographic scans were digitized, defining the axial morphology of the normal facet joint at five stations from proximal to distal within the joint. Assessments were made of facet joint orientation, transverse articular dimension, depth of the articular surface, and shape of the articular surface at levels L3-4, L4-5, and L5-S1. There was a gradually more coronal orientation from proximal to distal among the stations at each level, and a maximal transverse articular dimension at the level of the superior endplate of the caudad vertebra. Minimal error in the recording process at this level. In addition to the maximal joint dimension, made this level the most representative of the overall morphology and most useful for further studies. At the L4-5 level, a significantly more sagittal facet orientation was found in the degenerative spondylolisthesis group when compared to both the normal population and spinal stenosis groups (P < 0.01). At L5-S1, the only significant morphologic difference between the normal population and the patients with isthmic spondylolysis was reduced transverse articular dimension. These results support the hypothesis that patients developing degenerative spondylolisthesis are predisposed to this by a developmental sagittal orientation of the L4-5 facet joints.

Adult↗

N-Myristoylation and betagamma play roles beyond anchorage in the palmitoylation of the G protein alpha(o) subunit.

Many of the alpha subunits of heterotrimeric GTP-binding regulatory proteins (G proteins) are palmitoylated, a modification proposed to play a key role in the stable anchorage of the subunits to the plasma membrane. Palmitoylation of alpha subunits from the G(i) family is preceded by N-myristoylation, which alone or together with betagamma probably supports a reversible interaction of the alpha subunit with membrane as a prerequisite to the eventual incorporation of palmitate. Previous studies have not addressed, however, the question of whether membrane association alone, carried out through N-myristoylation, interaction with betagamma, or other events, is sufficient for palmitoylation. We report here for alpha(o) that it is not. We found that N-myristoylation is required for palmitoylation at least in part because it supports events subsequent to membrane attachment. Mutants of alpha(o) designed to target the subunit to membrane without an N-myristoyl group are unable to be palmitoylated as evaluated by incorporation of [(3)H]palmitate. Mutants of alpha(o) unable to interact normally with betagamma yet still attach to membrane demonstrate that betagamma, in contrast, is not required for palmitoylation. betagamma becomes necessary, however, when the N-myristoyl group is absent. Our results suggest that N-myristoylation and betagamma, while almost certainly relevant to the reversible interaction of alpha(o) with membrane, also play at least partly overlapping, post-anchorage roles in palmitoylation.

Adenosine Diphosphate Ribose↗

[Somatostatin in NRM plays roles in inducing analgesia and enhancing electroacupuncture analgesia].

The effects of microinjection of somatostatin (SS), somatostatin depletor cysteamine (CS) and anti-somatostatin serum (ASSS) on the pain threshold (PT) and electroacupuncture (EA) analgesia were investigated in the rat. The potassium iontophoresis induced tail-flick was used to measure the pain threshold. The "Zusanli" were stimulated with EA. The results indicated that microinjection of SS into necleus raphe magnus (NRM), could elevate PT and enhance EA analgesia. PT and EA analgesia were decreased when SS in NRM was depleted by CS or neutralized by ASSS. These data revealed that SS in NRM plays important roles in inducing analgesia and enhancing electroacupuncture analgesia.

Acupuncture Analgesia↗

RNA polymerase II elongation factors Spt4p and Spt5p play roles in transcription elongation by RNA polymerase I and rRNA processing.

Previous investigations into the mechanisms that control RNA Polymerase (Pol) I transcription have primarily focused on the process of transcription initiation, thus little is known regarding postinitiation steps in the transcription cycle. Spt4p and Spt5p are conserved throughout eukaryotes, and they affect elongation by Pol II. We have found that these two proteins copurify with Pol I and associate with the rDNA in vivo. Disruption of the gene for Spt4p resulted in a modest decrease in growth and rRNA synthesis rates at the permissive temperature, 30 degrees C. Furthermore, biochemical and EM analyses showed clear defects in rRNA processing. These data suggest that Spt4p, Spt5p, and, potentially, other regulators of Pol I transcription elongation play important roles in coupling rRNA transcription to its processing and ribosome assembly.

Chromosomal Proteins, Non-Histone↗

[Activities and authors. Role playing in health education].

One the forms that the capitalism met for perpetuate his ideology was through of scholastic institution and do this, your set doctrinaire. This space orderly frequently oppose teachers and students, where the practice is enforce through of foregoing scheme. At sphere of health, this scheme repeatedly, because the conception of health /disease nap works of health teach the subordinate population and the culture is transmit for health, basically on advice and norm for the people with process of culpability for theirs diseases. We would like at this work report our experience with the Pedagogic Psychodrama of ROMANA (1987), at raising your demands referring informations about Firsts Aids with teenagers matriculation on course for patrol at slum of Rocinha(R.J.), where the datas were analysis quality of discourse. The utilization of a methodology don't exclude the station context if subject of apprenticeship, contribute for understand your role and commitment with the transformation personal and collective, we hope contribute with Nursing, teach possibilities, pedagogic and creatives for humanization the Education at Health.

Adolescent↗

Myeloma--the integral role played by the professional nurse.

Myeloma is a malignancy of plasma cells which are terminally differentiated B-lymphocytes. The diagnosis may be made incidentally at routine blood testing, when an abnormality is found in the plasma proteins on electrophoresis. More usually the patients are symptomatic, with bone pain, anaemia, evidence of renal failure, or the metabolic abnormalities associated with increased plasma calcium and urate levels. Effective treatment will extend survival from 7 to approximately 30 months and at the same time improve the quality of life. Treatment is multidisciplinary, prominently involves the professional nurse and may arbitrarily be divided into two stages. Firstly, reversible lesions, such as dehydration and plasma hyperviscosity must be corrected, hypercalcaemia and hyperuricaemia improved and, if necessary, renal dialysis undertaken. Secondly, but of equal importance, is the need for specific therapy to be directed against the tumour itself, and both cytotoxic agents and irradiation have an important role to play. More recently, newer approaches have included high dose chemotherapy and bone marrow transplantation.

Combined Modality Therapy↗

Reactive oxygen species play roles on B cell surface receptor CD40-mediated proximal and distal signaling events: effects of an antioxidant, N-acetyl-L-cysteine treatment.

Reactive oxygen species (ROS) have been indicated as important signal mediators for many cell surface receptors. We previously demonstrated that ROS are generated by cross-linking surface receptor CD40 and consequently induce c-Jun N-terminal kinase activation and interleukin-6 secretion in murine B cells. In this study, we investigated further the involvement of ROS in CD40-mediated signaling events in B cells. CD40-mediated proximal events, which include protein serine phosphorylation, protein translocation between membranes and cytosol, as well as receptor complex formation, were inhibited after the pre-incubation of cells with an antioxidant N-acetyl-L-cysteine (NAC). Additionally, B cell responses after long-term ligation of CD40, such as protein expression, nuclear transcription factor kappaB (NFkappaB) activation, and cell proliferation, were also affected when cells were treated with NAC. These data suggest that CD40-induced ROS play critical roles in CD40-mediated B cell regulation.

Acetylcysteine↗

Current topics in pharmacological research on bone metabolism: Promyelotic leukemia zinc finger (PLZF) and tumor necrosis factor-alpha-stimulated gene 6 (TSG-6) identified by gene expression analysis play roles in the pathogenesis of ossification of the posterior longitudinal ligament.

To understand the molecular pathogenesis of ossification of the posterior longitudinal ligament of the spine (OPLL), an ectopic bone formation disease, we performed cDNA microarray analysis on cultured ligament cells from OPLL patients to understand the molecular pathogenesis of OPLL. We identified promyelotic leukemia zinc finger (PLZF) as one of up-regulated genes and tumor necrosis factor-alpha-stimulated gene 6 (TSG-6) as one of down-regulated gene during osteoblastic differentiation. We investigated the roles of PLZF in the regulation of osteoblastic differentiation of human mesenchymal stem cells (hMSCs) and C2C12 cells. siRNA-mediated gene-silencing of PLZF resulted in a reduction of the expression of osteoblast-specific genes such as the alkaline phosphatase, collagen 1A1, Runx2/CBFA1, and osteocalcin genes in the presence of osteogenic differentiation medium (OS) in hMSCs. The overexpression of PLZF induced CBFA1 induction, suggesting that PLZF is an upstream regulator of CBFA1 and thereby participates in promoting the ossification of spinal ligament cells in OPLL patients. Adenovirus-mediated TSG-6 overexpression in hMSCs resulted in suppression of osteoblastic differentiation induced by either BMP-2 or OS. TSG-6 can bind to BMP-2 directly and thereby could inhibit BMP-2 signaling. Taken together, these findings indicate that PLZF and TSG-6 play important roles in early osteoblastic differentiation.

Animals↗

Role-play gamers and National Guardsmen compared.

Scores of 54 fantasy role-game players and 64 National Guardsman were compared on a neuroticism scale and demographic variables. While the Role-gamers reported daydreaming and sleeping more than the Guardsmen, the popular stereotype that game players are withdrawn, emotionally immature adolescents was not confirmed. The typical game player was male with as many close friends as the guardsmen. Mean neuroticism scores did not appear to differ between the two groups and were not high enough to be considered clinically significant.

Adult↗

[Role playing and training for individual psychotherapy: some disadvantages to avoid].

The author first describes briefly the outcome of a session. He then indicates some positive aspects of this method : better understanding of the patient's attitudes ; desinhibition of the spontaneity of the psychotherapist and better self-understanding, increase in facility in gesture and nursing ; questioning of psychotheapeutic ideals not yet integrated. The second part of the communication is devoted to inconveniences to avoid : alienation in new group psychotherapeutic goals, increase to tendencies to "voyeurisme" and absence of analysis of transferential processes. The advantages of the method outrange the eventuel inconveniences, especially when it is possible resove them.

Education, Medical, Graduate↗

Lactose absorption and malabsorption in healthy Italian children: do the quantity of malabsorbed sugar and the small bowel transit time play roles in symptom production?

Using breath hydrogen analysis after an oral lactose load (2 g/kg; maximum 50 g), we investigated the prevalence of lactose malabsorption in 61 healthy Italian children aged 6-13 years. We also examined the relationship between symptoms and small bowel transit time and the degree of sugar malabsorption. Three of 61 subjects produced no H2 after both lactose and lactulose load and thus were eliminated at the outset. Lactose malabsorption was defined as excretion of greater than 20 ppm H2. Lactose intolerance was classified as mild (colicky pain, flatulence, abdominal distension, borborygmi) or severe (diarrhea). The frequency of lactose malabsorption in the children aged 6-8 years (group I) was 25%; in the children aged 8-11 years (group II), it was 35%, and in the children aged 11-13 years (group III) 56%. The differences in frequency between the first and the third groups were significant (p = 0.05). Three of 20 (15%) in group I, two of 20 (10%) in group II, and three of 18 (17%) in group III were classified not only as lactose malabsorbing, but also as lactose intolerant, with symptoms during and after the test. We found no difference in the small bowel transit times or in the quantities of malabsorbed lactose in symptomatic and asymptomatic malabsorbing subjects. Other factors that may play a role in symptom production are discussed.

Adolescent↗

Role play. Providing care to children and their families in the emergency center.

The collaborative model has many benefits for children and families, physicians, nurses, staff, and NPs. The model of care can be structured to charge lower fees for nonacute problems, thereby saving money. Various studies have reported savings of 20% to 38% in the cost of visits when patients were seen by NPs instead of physicians (Salkever, Skinner, Steinwach, & Katz, 1982). With appropriate triaging, waiting time may be reduced for children and families with nonacute problems. Perhaps the most important benefit is the teaching that the NP is able to provide for families in terms of managing particular health problems, safety needs, and preventive interventions and maintenance of wellness for children.

Child↗

Reactive oxygen species generated by hematopoietic cytokines play roles in activation of receptor-mediated signaling and in cell cycle progression.

Hematopoietic cytokines, including interleukin (IL)-3 and erythropoietin (Epo), regulate hematopoiesis by stimulating their receptors coupled with the Jak2 tyrosine kinase to induce receptor tyrosine phosphorylation and activate mainly the STAT5, PI3K/Akt, and Ras/MEK/ERK signaling pathways. Here we demonstrate that IL-3 or Epo induces a rapid and transient (peaking at 30 min) as well as late progressive increase in reactive oxygen species (ROS) in a hematopoietic progenitor model cell line, 32Dcl3, and its subclone expressing the Epo receptor (EpoR), 32D/EpoR-Wt. The cytokine-induced ROS generation was not affected in 32Dcl3 cells depleted of mitochondrial DNA. The antioxidant N-acetyl-L-cysteine (NAC) inhibited IL-3-induced tyrosine phosphorylation of Jak2, IL-3 receptor betac subunit (IL-3Rbetac), and STAT5 as well as activation-specific phosphorylation of Akt, MEK, and ERK, while treatment of cells with H2O2 activated these signaling events. NAC also inhibited the EpoR-induced transphosphorylation of IL-3Rbetac. Moreover, NAC treatment reduced the expression levels of c-Myc, Cyclin D2, and Cyclin E, and induced expression of p27, thus inhibiting the G1 to S phase transition of cells cultured with IL-3. Further studies have shown that the degradation of c-Myc was facilitated or inhibited by treatment of cells with NAC or H2O2, respectively. These data indicate that the rapid generation of ROS by cytokine stimulation, which is at least partly independent of mitochondria, may play a role in activation of Jak2 and the STAT5, PI3K/Akt, and Ras/MEK/ERK signaling pathways as well as in transactivation of cytokine receptors. The cytokine-induced ROS generation was also implicated in G1 to S progression, possibly through stabilization of c-Myc and induction of G1 phase Cyclin expression leading to suppression of p27.

Acetylcysteine↗

Role played by lactobacilli in controlling the population of vaginal pathogens.

The role of Lactobacillus species in the female urogenital tract as a barrier to infection is of considerable interest. These organisms are believed to contribute to the control of vaginal microbiota by competing with other microorganisms for adherence to epithelial cells and by producing antimicrobial compounds. These bactericidal compounds include organic acid, which lowers the vaginal pH, hydrogen peroxide, bacteriocin-like substances and possibly biosurfactants.

Antibiosis↗

The role played by cell-substrate interactions in the pathogenesis of osteoclast-mediated peri-implant osteolysis.

Prosthetic wear debris-induced peri-implant osteolysis is a major cause of aseptic loosening after total joint replacement. In this condition, wear particles released from the implant components induce a granulomatous inflammatory reaction at the interface between implant and adjacent bone, leading to progressive bone resorption and loss of fixation. The present study was undertaken to characterize definitively the phenotype of osteoclast-like cells associated with regions of peri-implant focal bone resorption and to compare the phenotypic features of these cells with those of mononucleated and multinucleated cells associated with polyethylene wear particles. Peri-implant tissues were obtained from patients undergoing hip revision surgery for aseptic loosening after total joint replacement. Cells were examined for the expression of several markers associated with the osteoclast phenotype using immunohistochemistry, histochemistry, and/or in situ hybridization. CD68 protein, a marker expressed by multiple macrophage lineage cell types, was detected in mononucleated and multinucleated cells associated with polyethylene particles and the bone surface. Cathepsin K and tartrate-resistant acid phosphatase were expressed highly in both mononucleated and multinucleated cells associated with the bone surface. Levels of expression were much lower in cells associated with polyethylene particles. High levels of beta3 integrin protein were detected in cells in contact with bone. Multinucleated cells associated with polyethylene particles exhibited faint positive staining. Calcitonin receptor mRNA expression was detected solely in multinucleated cells present in resorption lacunae on the bone surface and was absent in cells associated with polyethylene particles. Our findings provide further evidence that cells expressing the full repertoire of osteoclast phenotypic markers are involved in the pathogenesis of peri-implant osteolysis after total joint replacement. They also demonstrate that foreign body giant cells, although believed to be phenotypically and functionally distinct from osteoclasts, express many osteoclast-associated genes and gene products. However, the levels and patterns of expression of these genes in the two cell types differ. We speculate that, in addition to the role of cytokines and growth factors, the substrate with which these cells interact plays a critical role in their differential phenotypic and functional properties.

Acid Phosphatase↗