ObjectivesThe Endocuff is a device mounted on the tip of the colonoscope to help flatten the colonic folds during withdrawal. This study aimed to compare the adenoma detection rates between Endocuff-assisted (EC) colonoscopy and standard colonoscopy (SC).MethodsThis randomized prospective multicenter trial was conducted at four academic endoscopy units in Germany. Participants: 500 patients (235 males, median age 64[IQR 54–73]) for colon adenoma detection purposes were included in the study. All patients were either allocated to EC or SC. The primary outcome measure was the determination of the adenoma detection rates (ADR).ResultsThe ADR significantly increased with the use of the Endocuff compared to standard colonoscopy (35.4%[95% confidence interval{CI} 29–41%] vs. 20.7%[95%CI 15–26%], p<0.0001). Significantly more sessile polyps were detected by EC. Overall procedure time and withdrawal time did not differ. Caecal and ileum intubation rates were similar. No major adverse events occurred in both groups. In multivariate analysis, age (odds ratio [OR] 1.03; 95%[CI] 1.01–1.05), male sex (OR 1.74; 95%CI 1.10–2.73), withdrawal time (OR 1.16; 95%CI 1.05–1.30), procedure time (OR 1.07; 95%CI 1.04–1.10), colon cleanliness (OR 0.60; 95%CI 0.39–0.94) and use of Endocuff (OR 2.09; 95%CI 1.34–3.27) were independent predictors of adenoma detection rates.ConclusionsEC increases the adenoma detection rate by 14.7%(95%CI 6.9–22.5%). EC is safe, effective, easy to handle and might reduce colorectal interval carcinomas.Trial RegistrationClinicalTrials.gov NCT02034929.

We have developed an in situ hybridization assay capable of detecting enteroviral RNA in myocardial cells, using molecularly cloned coxsackievirus B3 cDNA as a diagnostic probe. Because of the high degree of nucleic acid sequence homology among the numerous enteroviral serotypes, including the group A and B coxsackieviruses and the echoviruses, detection of these various agents commonly implicated in human viral heart disease is possible in a single hybridization assay. We demonstrate the considerable potential of this method for an unequivocal diagnosis of enteroviral heart disease as well as for pathogenicity studies. Using athymic mice persistently infected with coxsackievirus B3 as a model system, we show that the myocardium is affected in a disseminated, multifocal manner.In North America and Europe, acute myocarditis is most commonly associated with infections by coxsackie B viruses (types 1 to 5). Other members of the human enteroviruses comprising at present over 70 serotypes (e.g., various coxsackie A viruses and echoviruses) are also considered to be relatively frequent causes of human viral heart disease (1-4). These agents appear to be capable of producing dilated cardiomyopathy of acute onset or lead to a variety of cardiac arrhythmias. Some of the acute cases may also evolve into a chronic form of dilated cardiomyopathy.The difficulty of establishing a specific diagnosis of viral heart disease is a major problem in clinical cardiology. Confirmation ofthe clinical suspicion of viral heart disease demands demonstration of replicating virus inside myocardial cells, which is exceedingly difficult by conventional methods (5). In addition, the increasing availability of potential antiviral agents has accentuated the need for methods by which endomyocardial biopsy specimens of patients with suspected viral heart disease can be diagnosed conclusively (6, 7).In order to introduce in situ nucleic acid hybridization (8-10) as a diagnostic tool in suspected enteroviral heart disease, we have recently cloned the single-stranded RNA genome of coxsackievirus B3 (CVB3) (11) that had been propagated in cultured human heart cells (12). Full-length reverse-transcribed cloned viral cDNA generated infectious CVB3 upon transfection into mammalian cells, demonstrating the molecular cloning of a faithful transcript ofthe original viral RNA. We now report the use of radioactively labeled cloned CVB3 cDNA as a diagnostic probe for the in situ detection of viral RNA in infected cultured cells and in myocardial tissue sections of CVB3-infected T-cell-deficient mice. We show that the molecular hybridization approach permits the detection of infected myocardial cells at the single-cell level, thus providing a unique possibility for an unequivocal diagnosis.A further advantage of the nucleic acid hybridization approach is provided by the high degree of genetic homology shared among the different serotypes of the human enterovirus group (11, 13-17), making possible their detection in a single hybridization assay. Detectio...

The human lipoprotein lipase gene was cloned and characterized. It is composed of 10 exons spanning -30 kilobases. The first exon encodes the 5'-untranslated region, the signal peptide plus the first two amino acids of the mature protein. The next eight exons encode the remaining 446 amino acids, and the tenth exon encodes the long 3'-untranslated region of 1948 nucleotides. The lipoprotein lipase transcription start site and the sequence of the 5'-flanking region were also determined. We compared the organization of genes for lipoprotein lipase, hepatic lipase, pancreatic lipase, and Drosophila yolk protein 1, which are members of a family of related genes. A model for the evolution of the lipase gene family is presented that involves multiple rounds of gene duplication plus exon-shuffling and intron-loss events.

A full-length reverse-transcribed, infectious cDNA copy of coxsackievirus B3 (CVB3) was used to determine the nucleotide sequence of this cardiotropic enterovirus. Comparison of the nucleotide sequence and the deduced amino acid sequence of the viral precursor polyprotein with the sequences of other group B coxsackieviruses (CVB1 and CVB4) demonstrates a high degree of genetic identity. They share about 80% homology at the nucleotide level and about 90% when the amino acid sequences of the polyproteins are compared. The potential processing sites of the coxsackievirus polyproteins, as deduced from alignment with the poliovirus sequence, are conserved among these enteroviruses with the exception of the cleavage sites between VP1 and 2APro and between polypeptides 2B and 2C. Comparison of the 5' termini of the enteroviral genomes reveals a high degree of identity, including the initial 5' consensus UUAAAACAGC, suggesting essential functions in virus replication. An important finding concerning the molecular basis of infectivity was that both recombinant CVB3 cDNA and in vitro-synthesized CVB3 RNA transcripts are infectious, although two initial 5' uridine residues found on the authentic CVB3 RNA were missing. Here, we report that cDNA-generated CVB3, as well as CVB3 generated by in vitro-synthesized RNA transcripts, regains the authentic initial 5' uridine residues during replication in transfected cells, indicating that the picornaviral primer molecule VPg-pUpU may be uridylylated in a template-independent fashion. The generation of virus or virus mutants with infectious recombinant CVB3 cDNA and in vitro-synthesized infectious CVB3 transcripts should provide a valuable means for studying the molecular basis of the pathogenicity of this cardiotropic enterovirus.

Lysosomal acid lipase (LAL) is a hydrolase essential for the intracellular degradation of cholesteryl esters and triacylglycerols. This report describes a multi-step procedure for the purification of LAL from human liver. After solubilization with non-ionic detergent, acid hydrolase activity was purified 17 000-fold to apparent homogeneity by sequential chromatography on Concanavalin A Sepharose, carboxymethyl-cellulose, phenyl Superose, Mono S cation exchange and Superose 12 gel-filtration columns. This procedure yielded two silver-staining protein bands of 56 kDa and 41 kDa on SDSPAGE. Size-exclusion chromatography of the 41-kDa protein indicated that the enzyme was catalytically competent as a monomer of approximately 38 kDa. When assayed in the presence of cholesteryl oleate or trioleoylglycerol, purified acid lipase had V,,, values of 4390 nmol fatty acid . min-' . mg protein and 4756 nmol fatty acid . min-' . mg protein-', and apparent K,,, values of 0.142 mM and 0.138 mh4, respectively. The purified enzyme was most active at low pH (4.5 -5.0) and required non-ionic detergent and ethylene glycol for optimal stability. Incubation of the 41-kDa acid lipase with endoglucosaminidase H reduced the molecular mass by 4-6 kDa, demonstrating Asn-linked glycosylation with high-mannose oligosaccharides. Deglycosylation did not affect enzymic activity, indicating that carbohydrates are not required for LAL activity. Based on partial peptide sequence, an oligonucleotide was synthesized and utilized to isolate LAL cDNA clones from a human liver cDNA library. A full-length LAL cDNA contained 2626 nucleotides and coded for a predicted protein of 372 amino acids, preceded by a 27 residue hydrophobic signal peptide. Hepatic LAL differed from fibroblast acid lipase at the N-terminus and revealed extensive similarities with human gastric lipase and rat lingual lipase, confirming a gene family of acid lipases. Northern hybridization using the complete LAL cDNA as a radiolabeled probe indicated striking differences in mRNA expression among human tissues. LAL mRNA was most abundant in brain, lung, kidney and mammary gland. Placenta and HeLa cells expressed intermediate amounts of LAL mRNA, while RNA extracted from liver and heart showed low levels of expression.Lysosomal acid lipase (LAL) is a lipolytic enzyme involved in the intracellular metabolism of cholesteryl esters and triacylglycerols derived from plasma lipoproteins [ 11. LAL is synthesized in a variety of cells, including fibroblasts, macrophages, lymphocytes and liver cells (reviewed in [2]). After synthesis at the endoplasmic reticulum, the enzyme is targeted to the lysosomal compartment [3]. In this subcellular location, it hydrolyzes cholesteryl esters and triacylglycerols, liberating cholesterol and free fatty acids. Cholesterol is transferred to the cytosol where it plays a central role in the cellular sterol homeostasis [4, 51. The physiological importance of LAL in the metabolism of neutral triacylglycerols and cholesteryl esters is illustrated by two inborn er...