No abstract

In a previous controlled trial we reported that 'gut directed' hypnotherapy appeared to be highly effective in the management of patients with severe refractory irritable bowel syndrome.' A criticism of this study has been that the follow up period of three months was too short to discount a placebo response to hypnosis.2' All 15 patients treated with hypnotherapy in the original study have now been followed up for over one year and these data are reported here together with experience on an additional 35 patients treated in a similar manner. Methods PATIENTSThe 15 patients receiving hypnotherapy in the original study have been followed up at three monthly intervals with an assessment of symptom scores and an additional session of hypnotherapy at each visit. The miean follow up period was 18 months (14-21 months). Patients were asked to telephone if they experienced a relapse so that an additional session of hypnotherapy could be arranged. A further 35 patients (31 women, four men, aged 23-65 years) with intractable irritable bowel syndrome have Address for correspondence: Dr P J Whorwell, Dept of Medicine, University of South Manchester, Nell Lane, West Didsbury, Manchester M20 8LR. Received for publication 11 July 1986. also been treated by hypnotherapy and followed for at least three months. These patients were divided into three groups. Group 1: classical cases of irritable bowel syndrome exhibiting abdominal pain, abdominal distension, and an abnormal bowel habit. Group 2: atypical cases lacking one or two of the three criteria necessary for Group 1. Group 3: patients with classical irritable bowel syndrome with significant psychopathology as judged by a score in excess of 14 on the 28 question general health questionnaire.4 The original study only included patients in Group 1.Hypnotherapy was carried out 1y PJW and consisted of half hour sessions of decreasing frequency over a three month period. Patients were given a tape for daily autohypnosis after the third session. Before hypnosis the patient was given a simple account of intestinal smooth muscle physiology. Hypnosis was induced by an eye fixation and arm levitation technique followed by standard deepening procedures. After general comments about improvement of health and wellbeing, attention was directed to the control of intestinal smooth muscle. The patient was asked to place his/her hand on the abdomen, feel a sense of warmth and relate this to asserting control over gut function. All sessions were concluded with standard ego strengthening suggestions. No subject was unable to be hypnotised. Ten weekly sessions of hypnotherapy were offered before treatment was 423 on 12 May 2018 by guest. Protected by copyright.

It has been claimed that hypnotherapy may influence a number of physiological parameters not readily amenable to conscious control,"4 and it is therefore possible that its effect in the irritable bowel syndrome results from a direct action on the gut. On the other hand, it is known that patients with the irritable bowel syndrome have a high incidence of psychopathology'5"6 and it may be that improvement during hypnotherapy is secondary to its psychotherapeutic potential.It was the purpose of the present study to perform a controlled investigation into the effect of hypnotherapy on the sensory and motor functions of the rectum in patients with irritable bowel syndrome.

Cellular cholesterol homeostasis is a fundamental and highly regulated process. Transcription factors known as sterol regulatory element binding proteins (SREBPs) coordinate the expression of many genes involved in the biosynthesis and uptake of cholesterol. Dysregulation of SREBP activation and cellular lipid accumulation has been associated with endoplasmic reticulum (ER) stress and activation of the unfolded protein response (UPR). This review will provide an overview of ER stress and the UPR as well as cholesterol homeostasis and SREBP regulation, with an emphasis on their interaction and biological relevance.

Renal proximal tubule injury is induced by agents/conditions known to cause endoplasmic reticulum (ER) stress, including cyclosporine A (CsA), an immunosuppressant drug with nephrotoxic effects. However, the underlying mechanism by which ER stress contributes to proximal tubule cell injury is not well understood. In this study, we report lipid accumulation, sterol regulatory element-binding protein-2 (SREBP-2) expression, and ER stress in proximal tubules of kidneys from mice treated with the classic ER stressor tunicamycin (Tm) or in human renal biopsy specimens showing CsA-induced nephrotoxicity. Colocalization of ER stress markers [78-kDa glucose regulated protein (GRP78), CHOP] with SREBP-2 expression and lipid accumulation was prominent within the proximal tubule cells exposed to Tm or CsA. Prolonged ER stress resulted in increased apoptotic cell death of lipid-enriched proximal tubule cells with colocalization of GRP78, SREBP-2, and Ca(2+)-independent phospholipase A(2) (iPLA(2)β), an SREBP-2 inducible gene with proapoptotic characteristics. In cultured HK-2 human proximal tubule cells, CsA- and Tm-induced ER stress caused lipid accumulation and SREBP-2 activation. Furthermore, overexpression of SREBP-2 or activation of endogenous SREBP-2 in HK-2 cells stimulated apoptosis. Inhibition of SREBP-2 activation with the site-1-serine protease inhibitor AEBSF prevented ER stress-induced lipid accumulation and apoptosis. Overexpression of the ER-resident chaperone GRP78 attenuated ER stress and inhibited CsA-induced SREBP-2 expression and lipid accumulation. In summary, our findings suggest that ER stress-induced SREBP-2 activation contributes to renal proximal tubule cell injury by dysregulating lipid homeostasis.

Chronic exposure of the skeletal muscle microcirculation to elevated shear stress-induces angiogenesis. Previous studies observed that shear stress-induced capillary growth involves luminal sprouting, or internal division, of the capillaries, which is characterized by a minimal proliferative response and the retention of an intact basement membrane. Matrix metalloproteinases (MMPs) are associated with the process of abluminal sprouting angiogenesis, but may not be required for the process of luminal division during capillary growth. We analyzed the production of MMP-2, using both the in vivo model of prazosin-induced angiogenesis in rat skeletal muscle, and cultured microvascular endothelial cells exposed to laminar shear stress. We found that MMP-2 was not elevated in capillaries of shear stress-stimulated skeletal muscle, despite a significant increase in capillary number in response to a shear stress stimulus. In cultured microvascular endothelial cells, MMP-2 mRNA and protein levels were attenuated significantly in response to shear stress exposure. This effect on MMP-2 was reversed by nitric oxide (NO) synthase inhibition using LNNA. In contrast, exposure of static cultures of endothelial cells to NO donors significantly reduced MMP-2 production. Shear stress exposure and NO donors both modified phosphorylation levels of several members of the MAPK family. Treatment of shear stress-exposed cells with the p38 MAPK inhibitor, SB203580, abolished the shear stress-mediated reduction in MMP-2 mRNA. Thus, our data provide strong evidence that elevated shear stress inhibits MMP-2 production in microvascular endothelial cells, an effect that is mediated by signal pathways involving both production of NO and activation of p38 MAPK.

Matrix metalloproteinase-2 (MMP-2) plays a critical role in endothelial cells during the processes of angiogenesis and vascular remodeling. Endothelial cell production of MMP-2 is greatly enhanced when cells are cultured within a three-dimensional type I collagen matrix coinciding with the increased invasive and migratory phenotype of the cells. To define the transcriptional regulation of MMP-2 in rat microvascular endothelial cells, we performed promoter-reporter assays with a series of promoter truncations. Activity of the full promoter was significantly greater in cells cultured within three-dimensional type I collagen compared with cells cultured as a monolayer (two-dimensional) on type I collagen. Truncation of the region encompassing base pairs ؊1562 to ؊1375 (relative to the start codon) of the MMP-2 promoter resulted in loss of this differential activity of the MMP-2 promoter. Analysis of this region indicated two putative GATA-2 binding domains between ؊1437 and ؊1387. Southwestern blot analysis and electrophoretic mobility shift assays confirmed the binding of GATA-2 to this region of the MMP-2 promoter. Overexpression of GATA-2 in COS-7 cells significantly increased the activity of the full-length MMP-2 promoter-luciferase construct. Endothelial cells expressed greater levels of GATA-2 protein in three-dimensional compared with two-dimensional cultures, and activity of the ؊1437/؊1387 region of the MMP-2 promoter was significantly greater in three-dimensional cultured endothelial cells. Together, these results indicate GATA-2 regulation of the MMP-2 promoter in endothelial cells and that the GATA-2 binding domain is sufficient to drive increased activity of the MMP-2 promoter in response to an extracellular matrix stimulus. The matrix metalloproteinases (MMPs)1 consist of a family of zinc-and calcium-dependent endopeptidases that cleave specific subsets of extracellular matrix proteins, growth factors, and cell surface receptors (1,2). Endothelial cell production and activation of MMPs, including MMP-2, are critical for the process of angiogenesis (3,4). Angiogenesis in skeletal muscle may be initiated by growth factors, by hemodynamic forces, or by mechanical stretching forces (5). The angiogenic process requires that endothelial cells proteolyze their basement membrane and then migrate through the interstitial matrix to form a new capillary. Inhibition of MMP activity prevents basement membrane degradation and the process of capillary sprouting in activity-stimulated skeletal muscle (6). When cultured within a three-dimensional type I collagen matrix, endothelial cells gain an invasive and migratory phenotype including increased production of matrix metalloproteinases and enhanced cell motility similar to what is seen in vivo during capillary sprouting (7,8). Regulated transcription of MMP-2 is important in determining this invasive phenotype in endothelial cells.Regulation of MMP-2 occurs at both transcriptional and post-translational levels. Although the rate of MMP-2 protein production is control...

Endoplasmic reticulum (ER) stress causes macrophage cell death within advanced atherosclerotic lesions, thereby contributing to necrotic core formation and increasing the risk of atherothrombotic disease. However, unlike in advanced lesions, the appearance of dead/apoptotic macrophages in early lesions is less prominent. Given that activation of the unfolded protein response (UPR) is detected in early lesion-resident macrophages and can enhance cell survival against ER stress, we investigated whether UPR activation occurs after monocyte to macrophage differentiation and confers a cytoprotective advantage to the macrophage. Human peripheral blood monocytes were treated with monocyte colony-stimulating factor to induce macrophage differentiation, as assessed by changes in ultrastructure and scavenger receptor expression. UPR markers, including GRP78, GRP94, and spliced XBP-1, were induced after macrophage differentiation and occurred after a significant increase in de novo protein synthesis. UPR activation after differentiation reduced macrophage cell death by ER stress-inducing agents. Further, GRP78 overexpression in macrophages was sufficient to reduce ER stress-induced cell death. Consistent with these in vitro findings, UPR activation was observed in viable lesion-resident macrophages from human carotid arteries and from the aortas of apoE(-/-) mice. However, no evidence of apoptosis was observed in early lesion-resident macrophages from the aortas of apoE(-/-) mice. Thus, our findings that UPR activation occurs during macrophage differentiation and is cytoprotective against ER stress-inducing agents suggest an important cellular mechanism for macrophage survival within early atherosclerotic lesions.