Polyadenylation promotes degradation of 3'-structured RNA by the Escherichia coli mRNA degradosome in vitro

Polyadenylation contributes to the destabilization of bacterial mRNA. We have investigated the role of polyadenylation in the degradation of RNA by the purified Escherichia coli degradosome in vitro. RNA molecules with 3'-ends incorporated into a stable stem-loop structure could not readily be degraded by purified polynucleotide phosphorylase or by the degradosome, even though the degradosome contains active RhlB helicase which normally facilitates degradation of structured RNA. The exoribonucleolytic activity of the degradosome was due to polynucleotide phosphorylase, rather than the recently reported exonucleolytic activity exhibited by a purified fragment of RNase E (Huang, H., Liao, J., and Cohen, S. N. (1998) Nature 391, 99-102). Addition of a 3'-poly(A) tail stimulated degradation by the degradosome. As few as 5 adenosine residues were sufficient to achieve this stimulation, and generic sequences were equally effective. The data show that the degradosome requires a single-stranded "toehold" 3' to a secondary structure to recognize and degrade the RNA molecule efficiently; polyadenylation can provide this single-stranded 3'-end. Significantly, oligo(G) and oligo(U) tails were unable to stimulate degradation; for oligo(G), at least, this is probably due to the formation of a G quartet structure which makes the 3'-end inaccessible. The inaccessibility of 3'-oligo(U) sequences is likely to have a role in stabilization of RNA molecules generated by Rho-independent terminators.     

The composition of milk fat.

Bovine milk contains about 3.5 to 5% total lipid, existing as emulsified globules 2 to 4 microns in diameter and coated with a membrane derived from the secreting cell. In homogenized milk, the coat is mostly casein. About 98% or more of the lipid is triacylglycerol, which is found in the globule. Phospholipids are about .5 to 1% of total lipids, and sterols are .2 to .5%. These are mostly located in the globule membrane. Cholesterol is the major sterol at 10 to 20 mg/dl. Data are presented on the membrane and the emulsion. Bovine milk contains substantial quantities of C4:0 to C10:0, about 2% each of C18:2 and trans-C18:1, and almost no other long-chain polyunsaturated fatty acids. The fatty acid composition is not altered by ordinary changes in diet. The triacylglycerol structure is unique, with much of the C4:0 to C10:0 at sn-3. The effects of milk cholesterol and fatty acids on human blood cholesterol levels and nonnutritive roles of some microlipids are discussed.     

Altruism on American Television: Examining the Amount of, and Context Surrounding, Acts of Helping and Sharing

Using a representative sample of television content featuring 2,227 programs across different genres and 18 different channels, the frequency and context of altruistic actions were content analyzed. A social cognitive theory approach was taken to guide the selection of contextual variables. The results showed that 73% of the programs in the sample featured instances of helping/sharing at a rate of 2.92 incidents per hour. Further, the actions were most likely to be initiated by adult White males and many acts were depicted in a humorous, realistic, and rewarding context. In terms of channel differences, shows on children's basic cable featured not only the highest proportion of programs with one or more instances of altruism but also the highest rate per hour (4.02 acts). Differences in contextual features and channel type are discussed in terms of social cognitive theory below.     

Perimetric motion thresholds are elevated in glaucoma suspects and glaucoma patients

The purpose of this study was to determine if a clinically feasible perimetric motion test utilizing random-dot kinematograms could identify glaucomatous visual field defects. Using a staircase procedure, an automated perimetric motion test and a larger foveally presented target were given to normal (n = 30), glaucoma suspects (n = 31) and primary open-angle glaucoma patients (n = 19). Motion thresholds at specific locations throughout the whole visual field were significantly elevated in glaucoma patients (P < or = 0.001). Perimetric motion testing identified 84.2% of the primary open-angle glaucoma patients and 25.8% of the glaucoma suspects as abnormal. A larger foveal stimulus was unable to distinguish between the different subject groups (P < or = 0.185). Perimetric motion thresholds were significantly correlated with Humphrey standard visual field thresholds in the glaucoma and glaucoma-suspect patients (P < or = 0.0002).     

Environmental health hazards: how children are different from adults

In policymaking on environmental health, it is often assumed that the entire population is exposed to and reacts to environmental contaminants in a similar manner. However, this assumption is misguided, especially where children are concerned. This article presents the scientific basis for the impacts of the environment on children, showing how children are different from adults in the ways in which they are exposed to environmental contamination and the ways in which they react to it when exposed. Specifically, the article examines the changing physical and biological environments of children. Children at different stages of development have unique physical risk factors for certain types of exposure because of changing location, levels of mobility, oxygen consumption, eating patterns, and behavior. When children are exposed to contaminants, their developing biological makeup--the way in which they absorb, distribute, and metabolize chemicals--will also affect how their bodies deal with the foreign substance. Each of these factors, along with the customs, laws, and regulations that affect the way in which children are exposed to the contaminants, had implications for the well-being of children in the years to come.     

Effect of topical butyrate on rectal epithelial kinetics and mucosal enzyme activities

Public awareness and misunderstandings of lactose intolerance are at an all-time high. Many people erroneously believe they are lactose intolerant or develop gastrointestinal symptoms after intake of lactose. Consequently, lactose-containing foods such as milk and other dairy foods may be eliminated unnecessarily from the diet. Because these foods are a major source of calcium, low intake of them can compromise calcium nutriture. This, in turn, can increase the risk of major chronic diseases such as osteoporosis (porous bones) and hypertension. This review is intended to help dietetics professionals alleviate clients' fears about lactose intolerance and recommend dietary strategies to improve tolerance to lactose. Scientific findings indicate that the prevalence of lactose intolerance is grossly overestimated. Other physiologic and psychologic factors can contribute to gastrointestinal symptoms that mimic lactose intolerance. Scientific findings also indicate that people with laboratory-confirmed low levels of the enzyme lactase can consume 1 serving of milk with a meal or 2 servings of milk per day in divided doses at breakfast and dinner without experiencing symptoms. Several dietary strategies are available to help lactose maldigesters include milk and other dairy foods in their diet without experiencing symptoms.     

No evidence for linkage of a novel CA repeat polymorphism for apoptosis antigen 1 (APO-1 or fas) with type I diabetes in a Caucasian population

Abstract DNA typing of four tetrameric repeat loci (HUMVWA, HUMTH0I, D21SII and HPRT) was carried out in a Chinese Han population from Shanghai (East China) and one from Guangzhou (South-East China) using a quadruplex PCR amplification and detection of the fluorescent-labeled alleles on the ALF DNA sequencer. All loci were in accordance with Hardy-Weinberg equilibrium except for D21S11 in the Guangzhou population. A test for population differentiation showed no statistical difference in the allele frequency distribution between the two populations. Comparison of the allele frequency data with other Chinese Han populations from North and South-West China for the STR loci HUMVWA and HUMTH01 revealed heterogeneity between Northern Chinese Han and Southern Chinese Han, which is in accordance with previous studies on the basis of protein markers.     

Identification and purification of diphosphoinositol pentakisphosphate kinase, which synthesizes the inositol pyrophosphate bis(diphospho)inositol tetrakisphosphate

Diphosphoinositol pentakisphosphate (PP-IP5) and bis(diphospho)inositol tetrakisphosphate (bis-PP-IP4) were recently identified as inositol phosphates which possess pyrophosphate bonds. The molecular mechanisms that regulate the cellular levels of these compounds are not yet characterized. To pursue this question, we have previously purified an inositol hexakisphosphate (IP6) kinase from rat brain supernatants [Voglmaier, S. M., et al. (1996) Proc. Natl. Acad. Sci. U.S.A. 93, 4305-4310]. We now report the identification and purification of another novel kinase, diphosphoinositol pentakisphosphate (PP-IP5) kinase, which uses PP-IP5 as a substrate to form bis(diphospho)inositol tetrakisphosphate (bis-PP-IP4) in soluble fractions of rat forebrain. The purified protein, a monomer of 56 kDa, displays high affinity (Km = 0.7 microM) and selectivity for PP-IP5 as a substrate. The purified enzyme also can transfer a phosphate from bis-PP-IP4 to ADP to form ATP. This ATP synthase activity is an indication of the high phosphoryl group transfer potential of bis-PP-IP4 and may represent a physiological role for PP-IP5 and bis-PP-IP4.     

7Li nuclear magnetic resonance study for the determination of Li+ properties in neuroblastoma SH-SY5Y cells

Lithium has been used clinically in the treatment of manic depression. However, its pharmacologic mode of action remains unclear. Characteristics of Li+ interactions in red blood cells (RBCs) have been identified. We investigated Li+ interactions on human neuroblastoma SH-SY5Y cells by developing a novel 7Li NMR method that provided a clear estimation of the intra- and extracellular amounts of Li+ in the presence of the shift reagent thulium-1,4,7,10-tetrazacyclododecane-N,N',N',N'-tetramethylene phosphonate (HTmDOTP4-). The first-order rate constants of Li+ influx and efflux for perfused, agarose-embedded SH-SY5Y cells in the presence of 3 mM HTmDOTP4- were 0.055 +/- 0.006 (n = 4) and -0.025 +/- 0.006 min(-1) (n = 3), respectively. Significant increases in the rate constants of Li+ influx and efflux in the presence of 0.05 mM veratridine indicated the presence of Na+ channel-mediated Li+ transport in SH-SY5Y cells. 7Li NMR relaxation measurements showed that Li+ is immobilized more in human neuroblastoma SH-SY5Y cells than in human RBCs.