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This study focuses on a pan-disciplinary scheme that targeted high-achieving undergraduate students. Earlier research from the scheme argued that high achievers have discernibly different learning and personal development support needs. One of the most frequent self-reported challenges within this high-achieving group is perfectionism. This “problematizing” of perfectionism by high-achieving students may present a challenge for learning professionals. Often the traits associated with perfectionism have played a key role in previous academic successes, yet are also reportedly experienced as injurious to the student's overall well-being. A discourse of academic resilience was employed to restructure students' perceptions and subsequent management of perfectionism. Literature from the fields of psychology and education were synthesised to create a conceptualisation of perfectionism that contributed positively to student well-being. This study suggests that reframing perfectionism within a discourse of academ...

Allogeneic hematopoietic stem cell transplantation is associated with serious complications, and improvement of the overall clinical outcome of patients with hematological malignancies is necessary. During the last decades, posttransplant donor-derived adoptive cellular immunotherapeutic strategies have been progressively developed for the treatment of graft-versus-host disease (GvHD), infectious complications, and tumor relapses. To date, the common challenge of all these cell-based approaches is their implementation for clinical application. Establishing an appropriate manufacturing process, to guarantee safe and effective therapeutics with simultaneous consideration of economic requirements is one of the most critical hurdles. In this review, we will discuss the recent scientific findings, clinical experiences, and technological advances for cell processing toward the application of mesenchymal stromal cells as a therapy for treatment of severe GvHD, virus-specific T cells for targeting life-threating infections, and of chimeric antigen receptors-engineered T cells to treat relapsed leukemia.

MicroRNAs (miRNA) have emerged as central regulators of diverse biological processes and contribute to driving pathology in several diseases. Acute graft-versus-host disease (aGvHD) represents a major complication after allogeneic hematopoietic stem cell transplantation, caused by alloreactive donor T cells attacking host tissues leading to inflammation and tissue destruction. Changes in miRNA expression patterns occur during aGvHD, and we hypothesized that we could identify miRNA signatures in target tissues of aGvHD that may potentially help understand the underlying molecular pathology of the disease. We utilized a rat model of aGvHD with transplantation of fully MHC-mismatched T cell depleted bone marrow, followed by infusion of donor T cells. The expression pattern of 423 rat miRNAs was investigated in skin, gut, and lung tissues and intestinal T cells with the NanoString hybridization platform, in combination with validation by quantitative PCR. MHC-matched transplanted rats were included as controls. In the skin, upregulation of miR-34b and downregulation of miR-326 was observed, while in the intestines, we detected downregulation of miR-743b and a trend toward downregulation of miR-345-5p. Thus, tissue-specific expression patterns of miRNAs were observed. Neither miR-326 nor miR-743b has previously been associated with aGvHD. Moreover, we identified upregulation of miR-146a and miR-155 in skin tissue of rats suffering from aGvHD. Analysis of intestinal T cells indicated 23 miRNAs differentially regulated between aGvHD and controls. Two of these miRNAs were differentially expressed either in skin (miR-326) or in intestinal (miR-345-5p) tissue. Comparison of intestinal and peripheral blood T cells indicated common dysregulated expression of miR-99a, miR-223, miR-326, and miR-345-5p. Analysis of predicted gene targets for these miRNAs indicated potential targeting of an inflammatory network both in skin and in the intestines that may further regulate inflammatory cytokine production. In conclusion, comprehensive miRNA profiling in rats suffering from aGvHD demonstrate tissue-specific differences in the expression patterns of miRNA that may not be detected by profiling of peripheral blood T cells alone. These tissue-specific miRNAs may contribute to distinct pathologic mechanisms and could represent potential targets for therapy. peerReviewed

For over 60 years, hematopoietic stem cell transplantation has been the major curative therapy for several hematological and genetic disorders, but its efficacy is limited by the secondary disease called graft versus host disease (GvHD). Huge advances have been made in successful transplantation in order to improve patient quality of life, and yet, complete success is hard to achieve. This review assimilates recent updates on pathophysiology of GvHD, prophylaxis and treatment of GvHD-related complications, and advances in the potential treatment of GvHD.

The reported inquiry-based learning (IBL) study was designed in 2012–2013 for the highest achieving undergraduate students at a research-intensive university in the United Kingdom (U.K.). In 2005, the University received national funding from the U.K. Higher Education Academy (HEA) to develop an innovative model of IBL to be used in a multidisciplinary context (Tosey, 2006). As a consequence, IBL was an obvious tool when, in 2012, the authors set out to design learning interventions to improve the teamwork and leadership skills of high-attaining students. In the process of exploring the application of IBL to this task, the need to ensure the intervention allowed for development in the conative domain was considered important. Historically, IBL practice at the University had catered well for cognitive and affective learning but had not been focussed to develop conation. A conative-heavy element was therefore purposefully designed into the latest IBL intervention.

Researchers who advocate the use of multiple methods often write interchangeably about ‘integrating’, ‘combining’ and ‘mixing’ methods, sometimes eliding these descriptors with ‘triangulation’, which itself encompasses several meanings. In this article we argue that such an elision is problematic since it obscures the difference between (a) the processes by which methods (or data) are brought into relationship with each other (combined, integrated, mixed) and (b) the claims made for the epistemological status of the resulting knowledge. Drawing on the literature for examples, we set out different rationales for using more than one method, then we develop a definition of integration of methods as a specific kind of relationship among methods. We also discuss different places in the research process where integration can occur: for instance, data from different sources can be integrated in the analysis stage, or findings from different sources at the point of theorizing.

The Editor's Corner appears periodically in Coronary Artery Disease. The Editor and Associate Editors consider ethical, social, and philosophical issues affecting cardiology and medicine.The personal relationship between one of the University of Vermont's recently graduated medical students and Dr D

Cyclic nucleotide-dependent vascular relaxation is associated with increases in the phosphorylation of a small heat shock protein (HSP), HSP20. An increase in phosphorylation of another small HSP, HSP27, is associated with impaired cyclic nucleotide-dependent vascular relaxation. Expression of HSPs is altered by exposure to several types of cellular stress in vitro. To determine if behavioral stress in vivo alters vascular expression and phosphorylation of the small HSPs and cyclic nucleotide-dependent vascular relaxation, borderline hypertensive rats were stressed by restraint and exposure to air-jet stress 2 h/day for 10 days or remained in their home cage. Stress impaired relaxation of aorta to forskolin, which activates adenylyl cyclase, and sodium nitroprusside, which activates guanylyl cyclase. This was associated with an increase in the aortic expression and phosphorylation of HSP27, which was localized to the vascular smooth muscle, but a decrease in the amount of phosphorylated (P)-HSP20. To determine if P-HSP27 inhibits phosphorylation of HSP20, P-HSP27 was added to a reaction mixture containing recombinant HSP20 and the catalytic subunit of cAMP-dependent protein kinase. P-HSP27 inhibited phosphorylation of HSP20 in a concentration-dependent manner. These data demonstrate that P-HSP27 can inhibit phosphorylation of HSP20. The increase in P-HSP27 and decrease in P-HSP20 were associated with reduced cyclic nucleotide-dependent vascular smooth muscle relaxation in response to behavioral stress in vivo, an effect similar to that observed previously in response to cellular stress in vitro.

For over a decade, a number of employers have been sounding warnings to the higher education sector that a “skills gap” was emerging at the employer/graduate interface. Some graduate recruiters no longer perceived academic competence alone to be sufficient and began calling for higher education to produce graduates with highly developed and recognisable transferable skills. Increasingly the graduates of the late 1990s are being asked to display far more than subject‐specific knowledge and are asked to prove that this skills development activity has occurred, usually by demonstrating that they have taken additional responsibility for something or someone during their higher education experience. This work examines what higher education can do to facilitate the development of transferable, and especially managerial, skills amongst the undergraduate population using extra‐curricula schemes.

This paper examines an established Student Tutoring programme in the higher education sector. In particular it investigates the methods by which the development of key skills in participating Student Tutors may be assessed and examines the procedures available for a Student Tutoring scheme to gain accredited status. In light of the recognised deficit in certain transferable skills amongst graduates, this paper also aims to encourage administrators to take the tutoring framework a step further, giving selected student participants the opportunity to gain management experience through overseeing aspects of a tutoring scheme.

Purpose: Cellular stress has been shown to induce a group of proteins called heat shock proteins (HSPs). Recent evidence suggests that a group of small HSPs may modulate vascular smooth muscle contraction (HSP27) and/or relaxation (HSP20). In this investigation, we hypothesized that cellular stress would alter contraction and/or relaxation of intact vascular smooth muscles and would lead to changes in the induction and/or phosphorylation of the small HSPs. Methods: Bovine carotid arteries were obtained from an abattoir, and physiologic contractile responses were determined in a muscle bath. Phosphorylation state–specific antibodies were produced and characterized against HSP27. Phosphorylation events were determined with phosphorylation state–specific antibodies or whole-cell phosphorylation and two-dimensional gel electrophoresis. Results: Cellular stress induced by arsenite or heat shock did not alter basal tone or the magnitude of contractions induced by serotonin or high extracellular potassium chloride. However, cellular stress led to inhibition of forskolin and sodium nitroprusside–induced vasorelaxation. This impaired vasorelaxation was associated with increases in the phosphorylation of HSP27 and decreases in forskolin-induced phosphorylation of HSP20. Conclusion: Cellular stress, which leads to increases in the phosphorylation of HSP27, inhibits cyclic nucleotide-dependent vascular relaxation and cyclic nucleotide-dependent increases in the phosphorylation of HSP20. (J Vasc Surg 2000;31:343-53.)

Cyclic nucleotide-dependent vasorelaxation is associated with increases in the phosphorylation of a small heat shock-related protein, HSP20. We hypothesized that phosphorylation of HSP20 in vascular smooth muscles is associated with alterations in the macromolecular associations of HSP20. Treatment of bovine carotid artery smooth muscles with the phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthine, and the adenylate cyclase activator, forskolin, led to increases in the phosphorylation of HSP20 and dissociation of macromolecular aggregates of HSP20. However, 3-isobutyl-1-methylxanthine and forskolin treatment of a muscle that is uniquely refractory to cyclic nucleotide-dependent vasorelaxation, human umbilical artery smooth muscle, did not result in increases in the phosphorylation of HSP20 or to dissociation of macromolecular aggregates. HSP20 can be phosphorylated in vitro by the catalytic subunit of cAMP-dependent protein kinase (PKA) in both carotid and umbilical arteries and this phosphorylation of HSP20 is associated with dissociation of macromolecular aggregates of HSP20. Activation of cyclic nucleotide-dependent signaling pathways does not lead to changes in the macromolecular associations of another small heat shock protein, HSP27. Interestingly, the myosin light chains (MLC20) are in similar fractions as the HSP20, and phosphorylation of HSP20 is associated with changes in the macromolecular associations of MLC20. These data suggest that increases in the phosphorylation of HSP20 are associated with changes in the macromolecular associations of HSP20. HSP20 may regulate vasorelaxation through a direct interaction with specific contractile regulatory proteins.

Postpartum vasospasm in the umbilical arteries may be due to impaired vasorelaxation secondary to alterations in the expression of heat shock proteins. The contractile responses of pre- and full-term bovine umbilical artery smooth muscles were determined in a muscle bath. Heat shock protein expression was determined in bovine and human arterial tissues using western blotting with specific antisera. Full-term bovine and human umbilical artery smooth muscle was refractory to relaxation induced by the nitric oxide donor, sodium nitroprusside. This impaired vasorelaxation was associated with the expression of the inducible form of the heat shock protein, HSP70i, and increases in the expression of the small heat shock protein, HSP27. Small heat shock proteins have been implicated in modulating contraction and relaxation responses in vascular smooth muscles. Thus, alterations in heat shock protein expression may play a role in umbilical artery vasospasm.

Human umbilical artery smooth muscle is uniquely refractory to cyclic nucleotide-dependent vasorelaxation. Small heat shock proteins (HSPs) have been implicated as contractile regulatory proteins. Thus, we hypothesized that alterations in the phosphorylation of small HSPs may contribute to human umbilical artery smooth muscle vasospasm. Physiologic contractile responses were determined in a muscle bath and compared with phosphorylation events determined with whole-cell phosphorylation and 2-dimensional gel electrophoresis. Precontraction of bovine carotid artery smooth muscle with serotonin followed by relaxation with forskolin was associated with increases in the phosphorylation of HSP27 and HSP20. Precontraction of umbilical artery with serotonin followed by forskolin treatment led to increases in the phosphorylation of HSP27. However, the umbilical artery smooth muscle did not relax, nor was there an increase in the phosphorylation of HSP20 with forskolin treatment. These data suggest that impaired cyclic nucleotide-dependent relaxation of umbilical artery smooth muscle is associated with a lack of phosphorylation of HSP20.

In the glomerular mesangial cell, platelet-derived growth factor (PDGF) activates several signal transduction pathways. We examined the effect of PDGF on cAMP production and on cAMP-dependent protein kinase (A-kinase) activation. In mesangial cells, PDGF stimulated cAMP production in a dose- and time-dependent manner. This effect of PDGF was not prevented by pre-incubation with 50 microM indomethicin. PDGF also activated type I A-kinase, the predominate A-kinase isoform in mesangial cells, measured either by a decrease in A-kinase photoaffinity labeling with 8-azido-[32P]-cAMP, or by an increase in A-kinase substrate phosphorylation. The activation of A-kinase by PDGF is not dependent on the intermediate production of prostaglandins or cGMP. These data suggest that A-kinase participates in PDGF-induced signaling events in mesangial cells.

Introduction: Suntinib malate (SM) is a small molecule tyrosine kinase inhibitor used for the treatment of metastatic renal cell carcinoma. However, nearly 20% of SM treated patients develop cardiomyopathy. We recently reported that the cardiomyocyte PDGFR-β regulates both cardiac and coronary microvascular function. In this study, we sought to define the mechanisms of SM-induced cardiomyopathy. Methods: C57BL/6 mice were divided into 4 groups: vehicle control (Ctrl), SM, Ctrl+transverse aortic constriction (Ctrl-TAC) and SM-TAC. SM was given at 40 mg/kg/d. An additional group was treated with either SM (40mg/kg/d) plus vehicle control or SM plus thalidomide (75mg/kg/d) for 14 days and allowed to recover until day 28. Left ventricular ejection fraction (LVEF) and coronary flow reserve (CFR, a measurement of coronary microvascular function) were assessed by cardiac MRI and ultrasound, respectively. Pericyte coverage was assessed by immunofluorescent co-staining of CD31 (vessel) and NG2 (pericyte). Results: SM induced cardiac and coronary microvascular dysfunction and impaired cardiac response to stress in a similar manner to that seen in PDGFR-β knockout mice. These functional impairments were accompanied by structural vascular defects and significant loss of microvascular pericyte coverage. This was recapitulated in aged (1 year) PDGFR-β knockout mice as well as with the drug CP673,541, which is a potent, more specific PDGFR inhibitor . Thalidomide is known to enhance vascular stability through enhancement of vascular pericyte coverage. Co-treatment with Thal prevented sunitinib-induced reduction in LVEF (SM-LVEF-41%; SM+thalidomide-49%, p Conclusion: SM-induced cardiac and microvascular dysfunction is most likely due to PDGFR-β inhibition and can be prevented by co-treatment with thalidomide. Our findings not only suggest a novel cardioprotective strategy for cancer patients at high-risk for sunitinib-induced cardiomyopathy but also suggest a critical role for pericytes in coronary microvascular and cardiac function.

Vascular diseases, such as atherosclerosis and restenosis following angioplasty or transplantation, are due to abnormal vascular smooth muscle growth and gene expression. The smooth muscle cells (SMC) in response to injury lose their contractile function, become highly proliferative and synthesize and secrete extracellular matrix proteins. Similar changes in the phenotypic properties of vascular SMC occur during in vitro culture. In this report, we examined whether restoration of the expression of the major receptor protein for nitric oxide (NO) signaling in smooth muscle, the guanosine 3′:5′ cyclic monophosphate (cGMP)-dependent protein kinase (PKG), reestablished contractile function to cultured rat aortic SMC. Contractile function was monitored using the silicone polymer wrinkle assay used previously to determine contractility in cultured mesangial cells. Noncontractile rat aortic smooth muscle cells transfected with the cDNA encoding the type I isoform of PKG, but not those transfected with empty vector, formed discreet wrinkles on the substratum in response to serum indicative of contraction. Treatment of the PKG-expressing SMC with sodium nitroprusside (SNP), an NO donor, and with cGMP analogs, or with the adenylyl cyclase activator, forskolin, and with adenosine 3′:5′ cyclic monophosphate (cAMP) analogs reduced wrinkling. The expression of a major PKG substrate protein involved in smooth muscle relaxation, heat shock-related protein-20 (HSP20), was also reestablished in PKG-expressing SMC. Treatment of the PKG-expressing SMC with nitroprusside resulted in phosphorylation of HSP20. Collectively, these results indicate that PKG expression is important to establish contractility to SMC in culture.

Background: Behavioral stress is a risk factor for hypertension and atherosclerosis. Stress leads to increases in the expression and phosphorylation of heat shock proteins (HSPs) in vascular smooth muscle. Two small heat shock proteins, HSP27 and HSP20, have been implicated in the regulation of the contraction and relaxation of smooth muscle. We hypothesized that stress-induced alterations in the phosphorylation of HSP27 would effect the macromolecular associations of the small HSPs. Methods: Bovine carotid artery smooth muscle was treated with buffer alone or with the chemical stressor, arsenite. HSP27 phosphorylation was determined with isoelectric focusing immunoblotting. Macromolecular interactions were determined with subcellular fractionation, molecular sieving, and glutaraldehyde cross-linking and immunoblotting. Results: Arsenite treatment led to increases in the phosphorylation of HSP27, a redistribution of some HSP27 from a cytosolic to a particulate fraction and to the formation of larger macromolecular aggregates of HSP27. Glutaraldehyde cross-linking and immunoblotting demonstrated that HSP27 existed in monomeric and dimeric forms, which suggested that the large aggregates were not simply aggregates of HSP27 but contained other proteins. Conclusions: Cellular stress leads to increases in the phosphorylation of HSP27 and to changes in the macromolecular associations of HSP27 in intact vascular smooth muscles. The functions of the small HSPs in the vascular smooth muscle may be dependent on both phosphorylation and macromolecular associations. (Surgery 2000;128:320-6.)

Infection with bacterial pathogens can disrupt the intestinal mucosa that maintains a healthy immune system while keeping bacteria at bay. Using a model of oral infection with a murinized form of Listeria monocytogenes (Lm), our lab has identified a γδ T cell population that forms bona-fide memory and provides protection from re-infection. Since the precise mechanism of activation of most γδ T cells has remained elusive, we investigated the requirements for antigen presenting cells (APCs) in the activation of Lm-induced memory γδ T cells. Here, we show that several cells in the mesenteric lymph node (mLN), including dendritic cells (DCs), inflammatory monocytes and granulocytes, can act in conjunction to activate γδ T cells to express IL-17A, IFN-γ or both and induced their proliferation. This activation was largely cell contact dependent, but was augmented by soluble factors at the peak of primary and secondary γδ T cell responses, mainly by IL-1β but partially by IL-2 and IL-23. Moreover, using confocal microscopy we were able to image Lm-induced γδ T cells interacting with DCs and inflammatory monocytes in the interfollicular area of the mLNs soon after Lm challenge infection. We believe these results not only advance the understanding of γδ T cell biology, but also provide novel strategies to design protective vaccines for the intestinal mucosa.

Four commercially available herpes simplex virus 1 (HSV-1) indirect fluorescent antibody (IFA) kits were compared with the use of sera selected because they were negative for HSV antibody by complement fixation (CF less than 1:8) and by ELISA (less than 1:100). However, 14 of 24 (58.3%) of these HSV-1 antibody-negative sera were positive at greater than or equal to 1:10 with the use of the HSV-1 IFA kit from Electronucleonics, 15 of 24 (62.5%) were positive with the Clinical Sciences HSV-1 IFA kit, 4 of 24 (16.7%) were positive with Zeus Scientific, and 4 of 18 (22.2%) were positive with the Gull Laboratories product. HSV-1 induces Fc receptors that commonly cause false positive IFA tests for HSV antibody. Therefore, further studies were undertaken to determine whether Fc receptors accounted for these false positive results. Staphylococcal protein A (SPA) is known to bind to the Fc portion of human IgG and therefore could be used to distinguish between the binding of an antibody by its Fab or its Fc portion. Thus, when fluorescein isothiocyanate conjugated (FITC) SPA was used as conjugate instead of FITC antibody to human IgG, true HSV-1 antibody-containing sera remained positive, but the false positives identified in the commercial IFA kits did not. The authors conclude that HSV-1-induced Fc receptors are responsible for most of the problem of these false positives and that HSV-1 serology probably should not be done by this type of IFA method until this problem is corrected.

Purpose: Thrombin mediates hemostasis by promoting thrombus development and vasospasm, which reduces the size of the arterial injury. Thrombin stimulation of vascular smooth muscle is associated with activation of mitogen-associated protein kinase. The purpose of this investigation was to determine the subsequent cellular signaling events in thrombin-stimulated vascular smooth muscle contraction. Methods: Contractile responses of bovine carotid artery smooth muscle were determined in a muscle bath and compared with phosphorylation events with two-dimensional gel electrophoresis. The activity of a novel kinase, mitogen-activated protein kinase-activated protein-2 kinase (MAPKAP2 kinase), was determined by immunoprecipitation and a phosphotransferase assay. A small heat shock protein, HSP27, was identified with immunoblotting. Results: Thrombin induces contraction of vascular smooth muscle and is associated with increased activity of MAPKAP2 kinase and increased phosphorylation of HSP27. Multiple isoforms of HSP27 are the predominant phosphoproteins in vascular smooth muscle, and peptide mapping suggests that the isoforms of HSP27 are structurally related and phosphorylated within similar peptide sequences. Conclusions: Activation of the MAPKAP2 kinase pathway and phosphorylation of HSP27 are associated with thrombin-induced contraction of vascular smooth muscle. (J Vasc Surg 1998;27:963-9.)