The conserved cellular roles of CLN proteins: Novel insights from Dictyostelium discoideum.

The neuronal ceroid lipofuscinoses (NCLs), collectively referred to as Batten disease, are a group of fatal neurodegenerative disorders that primarily affect children. The etiology of Batten disease is linked to mutations in 13 genes that encode distinct CLN proteins, whose functions have yet to be fully elucidated. The social amoeba Dictyostelium discoideum has been adopted as an efficient and powerful model system for studying the diverse cellular roles of CLN proteins. The genome of D. discoideum encodes several homologs of human CLN proteins, and a growing body of literature supports the conserved roles and networking of CLN proteins in D. discoideum and humans. In humans, CLN proteins have diverse cellular roles related to autophagy, signal transduction, lipid homeostasis, lysosomal ion homeostasis, and intracellular trafficking. Recent work also indicates that CLN proteins play an important role in protein secretion. Remarkably, many of these findings have found parallels in studies with D. discoideum. Accordingly, this review will highlight the translatable value of novel work with D. discoideum in the field of NCL research and propose further avenues of research using this biomedical model organism for studying the NCLs.

Malignant biliary disease: percutaneous interventions.

Interventional radiologists have an important role in the management of patients with malignant biliary obstruction. This article describes the techniques for percutaneous biliary drainage, insertion of biliary endoprostheses, and the management of occluded biliary endoprostheses. Most procedures are performed by using fluoroscopic guidance alone. Ultrasound is also a useful modality for guiding biliary drainage, particularly drainage of the left biliary ducts. Patients should be treated by internal drainage if possible. Metallic endoprostheses can be inserted at the time of the initial biliary drainage procedure. Plastic tubes should be inserted a few days after biliary drainage because of their relatively large size compared with metallic stents. Occluded plastic stents should be replaced. Blocked metallic stents should be treated either by placement of additional overlapping metallic stents or by placement of plastic stents within the metallic stent lumen.

A novel frameshift mutation in exon 23 of ATP7A (MNK) results in occipital horn syndrome and not in Menkes disease.

Menkes disease and occipital horn syndrome (OHS) are allelic, X-linked recessive copper-deficiency disorders resulting from mutations in ATP7A, or MNK. Classic Menkes disease has a severe phenotype, with death in early childhood, whereas OHS has a milder phenotype, with, mainly, connective-tissue abnormalities. Data suggest that steady-state localization of ATP7A to the trans-Golgi network (TGN) is necessary for proper activity of lysyl oxidase, which is the predominant cuproenzyme whose activity is deficient in OHS and which is essential for maintenance of connective-tissue integrity. Recently, it was reported that ATP7A-transcript levels as low as 2%-5% of normal are sufficient to result in the milder phenotype, OHS, rather than the phenotype of Menkes disease. In contrast to previously reported cases of OHS, we describe a case of OHS in which, because of a frameshift mutation, no normal ATP7A is produced. Although abundant levels of mutant transcript are present, there are substantially reduced levels of the truncated protein, which lacks the key dileucine motif L1487L1488. It has been demonstrated that the dileucine motif L1487L1488 functions as an endocytic signal for ATP7A cycling between the TGN and the plasma membrane. The present report is the first to describe an ATP7A truncation that results in OHS rather than in Menkes disease. The data from the present report support the concepts that (1) OHS results from lower levels of functional ATP7A and (2) ATP7A does not require the dileucine motif to function in copper efflux.

Internal stenting in malignant biliary obstruction.

Internal stenting in inoperable malignant biliary obstruction plays an important role in patient management. Surgical bypass may still be undertaken where there is also duodenal obstruction, though the need for gastroenterostomy may be reduced with the increasing use of metallic stents for the relief of malignant gastric outlet obstruction. Stents may be placed endoscopically or percutaneously, though in most centers the endoscopic route is usually tried first, with the percutaneous route being reserved for endoscopic failures. Plastic and self-expanding metallic biliary stents are available, each with its own advantages and disadvantages. In general, longer periods of patency are observed with metallic stents, though they are more expensive. Plastic stents can be changed endoscopically relatively easily when they have blocked, and in practice it is common for plastic stents to be inserted via this route for initial biliary drainage. If there is prolonged survival thereafter, many workers insert a metallic stent in an attempt to reduce the number of interventions required. If the percutaneous route is being employed, the histologic diagnosis has been confirmed, and the malignancy is inoperable, our practice is to use a self-expanding metallic stent, as the delivery system is relatively small and subsequent occlusion less likely.

Self-expanding metallic stents in the management of pyloric dysfunction after gastric pull-up operations.

The purpose of this paper is to report the use and benefits of self-expanding metallic stents employed in pyloric dysfunction. Four patients treated with oesophagectomy and gastric pull-up for oesophageal carcinoma failed to respond to balloon dilatation for pyloric dysfunction. Three of the patients were thought to have residual tumour at sites remote from the pylorus prior to stenting, but the fourth, who had undergone surgery 8 years previously, was thought to be cured. All were treated with self-expanding metallic stents. All four patients responded well with resolution of their symptoms. Over a mean follow-up of 6 months there has been no recurrence of symptoms. Stent insertion represents a potentially valuable method of treatment in patients with post-surgical pyloric dysfunction in whom simple balloon dilatation has failed.

The murine Fhit gene is highly similar to its human orthologue and maps to a common fragile site region.

The human FHIT gene is a putative tumor suppressor gene that maps to human chromosome band 3p14.2 in a region that is frequently deleted in cancers. It exhibits both genomic deletions and aberrant transcripts in a variety of tumors and spans the common fragile site FRA3B. This fragile site extends over a broad region of several hundred kb within the FHIT gene and may account for its instability in tumors. As one test of this hypothesis, we isolated the murine Fhit gene and asked whether it also contains a common fragile site and if it is unstable in mouse tumors or tumor cell lines. The Fhit gene was isolated, and the sequence was found to be 87.5% identical to that of the human FHIT gene in the open reading frame. Using fluorescence in situ hybridization, Fhit was assigned to mouse chromosome band 14A2, in a region that was previously shown to contain an aphidicolin-inducible mouse fragile site. Fluorescence in situ hybridization with genomic clones containing Fhit and flanking sequences demonstrated that gaps and breaks in the fragile site occur over a broad region within and proximal to the Fhit locus. Thus, the physical relationship of Fhit to a common fragile site is similar to that observed with the orthologous human FHIT gene and FRA3B.

Immunocytochemical localization of the Menkes copper transport protein (ATP7A) to the trans-Golgi network.

We have generated polyclonal antibodies against the amino-terminal third of the Menkes protein (ATP7A; MNK) by immunizing rabbits with a histidine-tagged MNK fusion construct containing metal-binding domains 1-4. The purified antibodies were used in Western analysis of cell lysates and in indirect immunofluorescence experiments on cultured cells. On Western blots, the antibodies recognized the approximately 165 kDa MNK protein in CHO cells and human fibroblasts. No MNK signal could be detected in fibroblasts from a patient with Menkes disease or in Hep3B hepatocellular carcinoma cells, confirming the specificity of the antibodies. Immunocytochemical analysis of CHO cells and human fibroblasts showed a distinct perinuclear signal corresponding to the pattern of the Golgi complex. This staining pattern was similar to that of alpha-mannosidase II which is a known resident enzyme of the Golgi complex. Using brefeldin A, a fungal inhibitor of protein secretion, we further demonstrated that the MNK protein is localized to the trans-Golgi network. This data provides direct evidence for a subcellular localization of the MNK protein which is similar to the proposed vacuolar localization of Ccc2p, the yeast homolog of MNK and WND (ATP7B), the Wilson disease gene product. In light of the proposed role of MNK both in subcellular copper trafficking and in copper efflux, these data suggest a model for how these two processes are linked and represent an important step in the functional analysis of the MNK protein.

Metallic endoprostheses for malignant tracheobronchial obstruction: initial experience.

PURPOSE: To assess the efficacy of the Wallstent endoprosthesis in malignant tracheobronchial obstruction. METHODS: Seven patients with irresectable carcinoma of the bronchus were treated with nine Wallstent endoprostheses. The procedures were performed under endoscopic and fluoroscopic guidance. Wallstent endoprostheses ranging from 8-16 mm in diameter and 26-49 mm in length were deployed after balloon dilatation of the strictures. RESULTS: All stents were successfully deployed in the desired positions. There was one procedural complication and one procedure related death. Three patients showed significant improvement in respiratory status after stenting. At a mean follow-up of 5.1 months, there has been no stent migration, fracture, or collapse. One patient had proximal tumor overgrowth that was treated with additional stent insertion. One patient died after a bout of massive hemoptysis 3 months poststenting and it was difficult to tell whether this was related to the endoprosthesis. CONCLUSION: The use of the Wallstent endoprosthesis in malignant tracheobronchial obstruction is technically feasible.

Short communication: tunnelled central venous catheter insertion via the internal jugular vein using a dedicated portable ultrasound device.

We describe the use of a purpose-built portable ultrasound device to assist puncture of the internal jugular vein during percutaneous insertion of tunnelled central venous catheters such as Hickman lines. In many situations the internal jugular route is safer, faster and less costly in comparison with the subclavian approach. The use of ultrasound allows an accurate initial venous puncture with fewer complications, in the hands of both experienced operators and those less familiar with the internal jugular vein approach.

Case report: persistence of lipiodol for 13 months in metastatic deposits in the liver on computed tomography.

The computed tomography (CT) findings in a 17-year-old male student with liver metastases from a primary adenocarcinoma of the rectum are presented. Lipiodol-enhanced CT 4 days after lipiodol infusion demonstrated several metastases not visible on conventional CT. A repeat CT 13 months later showed lipiodol to have been retained within the original lesions and also demonstrated new metastatic deposits free of lipiodol. Although persistence of lipiodol for up to a year has been reported by workers in Japan imaging hepatocellular carcinoma (Yumoto et al., 1985) this experience has not been confirmed in the UK (Raby et al., 1989). There are relatively few reports investigating metastatic adenocarcinoma (Nakakuma et al., 1985), none of which refers to persistence of lipiodol over 1 year. This case report demonstrates the ability of lipiodol-enhanced CT to detect occult metastases and also to facilitate their follow-up and differentiation from newer lesions.

Case report: vascular anomaly in Dieulafoy's disease.

We present a case of a vascular anomaly with Dieulafoy's disease in a young man. The diagnosis was made by coeliac and selective left gastric angiography after endoscopy, laparotomy and angiography had failed to detect the lesion. The patient was treated successfully by laparotomy and resection of the bleeding area in the stomach. This is the second report of a vascular anomaly with Dieulafoy's disease.