Expression, purification, reconstitution and inhibition of Ustilago maydis sterol 14 alpha-demethylase (CYP51; P450(14DM))

PURPOSE: To assess the efficacy of percutaneous embolization with an alcoholic solution of zein in the treatment of aneurysmal bone cysts. MATERIALS AND METHODS: Eighteen patients with aneurysmal bone cysts were treated percutaneously with alcoholic zein. The cysts were in the lower limb (n = 7), upper limb (n = 5), pelvis (n = 3), cervical spine (n = 2), and mandible (n = 1). All patients were symptomatic, three had previously undergone surgery. Percutaneous embolization was performed with fluoroscopic or computed tomographic guidance with the patient under general anesthesia. Clinical and imaging follow-up lasted 18 months to 4 years. RESULTS: Percutaneous embolization was performed in 16 cases. In two cases, cystograms showed marked venous drainage and thus embolization was not attempted. Six patients underwent repeat embolization. Complications consisted of a local transitory inflammatory reaction (n = 5), aseptic osteitis (n = 1), and a small pulmonary infarct without sequelae (n = 1). Relief of symptoms was achieved in all patients except one, who underwent surgery. At imaging, improvement was total in 13 cases (87%) and partial in two cases (13%). No recurrence was noted during follow-up. CONCLUSION: Percutaneous embolization of aneurysmal bone cysts with alcoholic zein should be considered a reliable alternative to surgery, especially in cases with a difficult surgical approach or cases of postsurgical recurrence.     

Measuring the effect of a hygiene behaviour intervention by indicators of behaviour and diarrhoeal disease

A social marketing approach used both qualitative and quantitative methods to develop a hygiene behaviour intervention in rural north-east Thailand. Behaviours were preselected from a previous study and the intervention was designed to promote hand washing, especially before feeding a baby, cooking, eating, and after defaecation or cleaning a baby's bottom, and dish washing immediately after eating. A bacteriological indicator (enumerating faecal streptococci using a finger impression technique) was developed to measure changes in hand washing behaviour and observation (spot checks) of dirty dishes to indicate dish washing practice. There was a significant improvement in both behaviours and a significant reduction in diarrhoeal disease as a result of the intervention. Furthermore, both indicators were retrospectively found to be positively related to diarrhoeal disease incidence. However, receiving and being able to recall the intervention messages was not necessarily sufficient to ensure behaviour change, as some adults found it difficult to change old habits. Villages showing the greatest improvement tended to have a stronger sense of community than others and to have more people actively involved in the intervention.     

An integrated analysis of the progression of cell responses induced by permanent focal middle cerebral artery occlusion in the rat

Defining the chronology and severity of cell damage in an evolving lesion after ischemia is important for understanding the underlying mechanisms in the development of therapeutic intervention. In the present study, we used a combination of histological and immunocytochemical methods to evaluate cell responses from 30 min to 48 h after permanent occlusion of the middle cerebral artery (MCAO) in the rat. Specific immunocytochemical markers clearly revealed acute early responses in neurons (neurofilament protein 200), astrocytes (glial fibrillary acidic protein), and microglia/macrophages (OX-42 and ED-1) such as enlarged, convoluted neuronal processes, and disintegration of glia. Progressive topographic changes in the developing lesion, pinpointed by immunolabeling, indicated the severity and extension of the cell damage. Proliferation and hypertrophy of astrocytes and microglia around the infarct, and contralaterally, occurred 24-48 h after MCAO and coincided with mass necrosis and infiltration of neutrophils and macrophages into the core. These observations corroborate the suggestion that the inflammatory process is involved in the progression of the infarct.     

Development of a two-dimensional gel electrophoresis database of human lysosomal proteins

Two-dimensional gel electrophoresis databases have been generated for a range of tissue cell and fluid types, from a number of species. A major difficulty in the development of such databases is the large number of proteins present in even a single cell type (> 10,000) and the low levels of many of these proteins. One approach to overcome these difficulties is to fractionate the cell into its organelles and generate individual databases for each subcellular component. This has the added advantage of assigning a cellular location to each protein identified. Here we report the development of a two-dimensional gel electrophoresis database of lysosomal proteins.     

Treatment of irritable bowel syndrome with Chinese herbal medicine: a randomized controlled trial

CONTEXT: Irritable bowel syndrome (IBS) is a common functional bowel disorder for which there is no reliable medical treatment. OBJECTIVE: To determine whether Chinese herbal medicine (CHM) is of any benefit in the treatment of IBS. DESIGN: Randomized, double-blind, placebo-controlled trial conducted during 1996 through 1997. SETTING: Patients were recruited through 2 teaching hospitals and 5 private practices of gastroenterologists, and received CHM in 3 Chinese herbal clinics. PATIENTS: A total of 116 patients who fulfilled the Rome criteria, an established standard for diagnosis of IBS. INTERVENTION: Patients were randomly allocated to 1 of 3 treatment groups: individualized Chinese herbal formulations (n = 38), a standard Chinese herbal formulation (n = 43), or placebo (n = 35). Patients received 5 capsules 3 times daily for 16 weeks and were evaluated regularly by a traditional Chinese herbalist and by a gastroenterologist. Patients, gastroenterologists, and herbalists were all blinded to treatment group. MAIN OUTCOME MEASURES: Change in total bowel symptom scale scores and global improvement assessed by patients and gastroenterologists and change in the degree of interference in life caused by IBS symptoms assessed by patients. RESULTS: Compared with patients in the placebo group, patients in the active treatment groups (standard and individualized CHM) had significant improvement in bowel symptom scores as rated by patients (P=.03) and by gastroenterologists (P=.001), and significant global improvement as rated by patients (P=.007) and by gastroenterologists (P=.002). Patients reported that treatment significantly reduced the degree of interference with life caused by IBS symptoms (P=.03). Chinese herbal formulations individually tailored to the patient proved no more effective than standard CHM treatment. On follow-up 14 weeks after completion of treatment, only the individualized CHM treatment group maintained improvement. CONCLUSION: Chinese herbal formulations appear to offer improvement in symptoms for some patients with IBS.     

Calpain activation is upstream of caspases in radiation-induced apoptosis

The molecular events involved in apoptosis induced by ionizing radiation remain unresolved. In this paper we show that the cleavage of fodrin to a 150 kDa fragment is an early proteolytic event in radiation-induced apoptosis in the Burkitts' Lymphoma cell line BL30A and requires 100 microM zVAD-fmk for inhibition. Caspases-1, -3, -6 and -7 were shown to cleave fodrin to the 150 kDa fragment in vitro and all were inhibited by 10 microM zVAD-fmk. We also show that the in vitro cleavage of fodrin by calpain is inhibited by 100 microM zVAD-fmk as was the calpain-mediated hydrolysis of casein. We demonstrate that calpain is activated within 15 min after radiation exposure, concomitant with the cleavage of fodrin to the 150 kDa fragment whereas caspase-3 is activated at 2 h correlating with the cleavage of fodrin to the 120 kDa fragment. These results support a role for calpain in the early phases of the radiation-induced apoptosis pathway, upstream of the caspases.     

United Kingdom Cancer Research Campaign approach to quality-of-life research in cancer clinical trials

Clinical trials of new anticancer therapies form an important part of the research activity of the Cancer Research Campaign (United Kingdom), and quality-of-life (QOL) end points are being increasingly used in the evaluation of new treatment approaches. The Campaign has a unique policy of supporting a broad range of scientific and clinical research, including psychosocial studies, and thus QOL research is generated in a variety of clinical settings. The focus of interest for the Cancer Research Campaign lies in QOL design and assessment rather than the routine application of QOL protocols. Clinical investigators are free to adopt in individual approach, but the Campaign operates a strict peer-review system in protocol assessment. Some standardization of approach is being achieved through consensus of opinion and wide collaboration, both nationally and internationally.