Polypharmacy and Crohn's disease.

BACKGROUND: Polypharmacy has not been defined for Crohn's disease. AIMS: To determine the prevalence of polypharmacy, factors associated with polypharmacy, and consequences of polypharmacy in a Crohn's disease population. METHODS: A review of 291 Crohn's disease patients was performed. Polypharmacy was defined as either minor (two to four medications) or major (> or = 5 medications). Clinical status was evaluated with the Harvey-Bradshaw index of disease activity (HBI) and the short inflammatory bowel disease questionnaire (SIBDQ). RESULTS: Major polypharmacy was identified in 50% of patients. Crohn's disease patients on less than two medications at the intake visit had an HBI of 3.6 compared with 5.4 and 6.0 in the minor and major polypharmacy groups (P < 0.05). Similarly, patients on less than two medications had an SIBDQ of 60.3 compared with 55.7 and 53.4 in the minor and major polypharmacy groups (P = 0.11). Predictors of polypharmacy included age > 40 years (OR 1.9), duration of disease > 10 years (OR 2.0), and female sex (OR 2.5). CONCLUSIONS: Polypharmacy is common in Crohn's disease and correlates with increased disease activity and decreased quality of life. Increasing age, increasing duration of disease, and female sex are associated with major polypharmacy. These findings emphasize the need for improved treatment algorithms to optimize Crohn's disease patient management.

Identification of a unique guanine-7-methyltransferase in Semliki Forest virus (SFV) infected cell extracts.

The methylation of the 5' terminal guanosine residue of the cap structure of Semliki Forest virus (SFV) mRNAs has been shown to occur in vitro concomitantly with their synthesis (R. K. Cross and P. J. Gomatos, Virology, 114, 542-554, 1981). The enzyme responsible for this methylation, a guanine-7-methyltransferase, is associated with the SFV replication complex which contains both the virus-specified polymerase and RNA template in a mitochondrial pellet fraction, P-15, from infected cell lysates. In the present report, evidence has been obtained demonstrating that a virus-specified function is required for this methylating activity. First, the methyltransferase enzyme in these infected P-15 extracts has been found to differ in substrate specificity from that of the BHK host cell enzyme. This enzyme was able to catalyze the methylation of GTP to m7GTP in vitro whereas the cellular enzyme could not methylate GTP. The incorporation of a methyl group onto GTP occurred linearly for at least 2 hr at 30 degrees under conditions of neutral pH and added GTP substrate. Second, a study of the kinetics of appearance of this activity, has demonstrated that the capacity to methylate GTP did not appear until 1 hr after infection and reached maximal levels by about 3 hr. Third, de novo protein synthesis was required. Addition of the protein synthesis inhibitor, cycloheximide, prevented the appearance and subsequent increase in the methylating activity. However, once formed the methyltransferase was found to be stable for at least 3 hr. These results suggest that an early viral function, perhaps a nonstructural polypeptide is required for this novel guanine-7-methyltransferase activity in SFV infected cell extracts.

Involvement of microtubules and 10-nm filaments in the movement and positioning of nuclei in syncytia.

Previous studies (Holmes, K.V., and P.W. Choppin. J. Exp. Med. 124:501-520; J. Cell Biol. 39:526-543) showed that infection of baby hamster kidney (BHK21-F) cells with the parainfluenza virus SV5 causes extensive cell fusion, that nuclei migrate in the syncytial cytoplasm and align in tightly-packed rows, and that microtubules are involved in nuclear movement and alignment. The role of microtubules, 10-nm filaments, and actin-containing microfilaments in this process has been investigated by immunofluorescence microscopy using specific antisera, time-lapse cinematography, and electron microscopy. During cell fusion, micro tubules and 10-nm filaments from many cells form large bundles which are localized between rows of nuclei. No organized bundles of actin fibers were detected in these areas, although actin fibers were observed in regions away from the aligned nuclei. Although colchicine disrupts microtubules and inhibits nuclear movement, cytochalasin B (CB; 20-50 microgram/ml) does not inhibit cell fusion or nuclear movement. However, CB alters the shape of the syncytium, resulting in long filamentous processes extending from a central region. When these processes from neighboring cells make contact, fusion occurs, and nuclei migrate through the channels which are formed. Electron and immunofluorescence microscopy reveal bundles of microtubules and 10-nm filaments in parallel arrays within these processes, but no bundles of microfilaments were detected. The effect of CB on the structural integrity of microfilaments at this high concentration (20 microgram/ml) was demonstrated by the disappearance of filaments interacting with heavy meromyosin. Cycloheximide (20 microgram/ml) inhibits protein synthesis but does not affect cell fusion, the formation of microtubules and 10-nm filament bundles, or nuclear migration and alignment; thus, continued protein synthesis is not required. The association of microtubules and 10-nm filaments with nuclear migration and alignment suggests that microtubules and 10-nm filaments are two components in a system which serves both cytoskeletal and force-generating functions in intracellular movement and position of nuclei.

Genome RNAs and polypeptides of reovirus serotypes 1, 2, and 3.

The virus-specific double-stranded genome RNA and polypeptides present in virions and cells infected with the three mammalian reovirus serotypes have been examined by co-electrophoresis in several different polyacrylamide gel systems. The double-stranded RNA and polypeptide species previously described for type 3 Dearing were found to have corresponding species in the other serotypes examined. In each serotype several RNA and polypeptide species were found to have different electrophoretic mobilities from the corresponding RNA or polypeptide species of type 3 Dearing. The combination of electrophoretic variants among the RNAs and polypeptides of the reovirus serotypes gave electrophoretic markers in all 10 of the reovirus genes. The usefulness of these electrophoretic markers in "mapping" the reovirus genome is discussed.

Temperature-sensitive mutants of reovirus type 3: evidence for aberrant mu 1 and mu 2 polypeptide species.

An analysis of reovirus-specific polypeptides in cells infected with temperature-sensitive mutants under permissive and nonpermissive conditions revealed the presence of (i) all the known viral polypeptides and (ii) aberrant migration of the mu 1 and mu 2 polypeptides in four groups of mutants.

Reovirus-specific polypeptides: analysis using discontinuous gel electrophoresis.

The electrophoretic analysis of reovirus-specific polypeptides in infected cells using a discontinuous gel system has allowed the resolution of additional viral-specific polypeptides, including one large-sized gamma3 and two (or possibly three) medium-sized (mu3, mu4, mu5(?)) species. The proteins designated mu0, sigma1, and sigma2 based on electrophoretic mobility in gel systems containing phosphate-urea correspond to mu4, sigma2, and sigma1, respectively, when analyzed in systems containing Tris-glycine. It is likely that protein modifications (phosphorylation and glycosylation) are responsible for at least some of these differences.