The current and future role of the Internet in patient education

The Internet technology known as the World Wide Web is rapidly emerging as the most powerful medium of mass communication this century and it can be harnessed to dispense global, cost-effective, high-quality, multimedia patient education material. This paper reviews how the Internet has progressed from delivering simple static, text-based material to sophisticated interactive Web sites based on CGI Technology. Interactive Web sites can be used to deliver health assessment questionnaires and Web-based decision-support systems can give patients advice on the emergency management of acute medical problems. The advantages and drawbacks of this new technology, including information regulation and quality are discussed. The role of the Hospital Intranet as a patient education resource is described. The paper concludes by illustrating how patients can appreciate the 3-D structure of bones and organs using virtual reality in a VRML Web environment.     

An evaluation of ruminally degradable intake protein and metabolizable amino acid requirements of feedlot calves

Ruminally degradable intake protein (DIP) and metabolizable indispensable amino acid (MIAA) requirements of feedlot steers were evaluated. Dietary treatments consisted of isocaloric 80% concentrate steam-flaked corn-based diets containing either .8% urea, 1.5% fish meal (FM), 3.0% FM, 4.5% FM, or 4.5% soybean meal (SBM). Treatment effects on characteristics of ruminal and total tract digestion were evaluated using four Holstein steers (249 kg) with cannulas in the rumen and proximal duodenum. Ruminal digestibility of OM (RDOM; P < .05) and feed N (P < .01) and microbial N flow (MNF; P < .01) to the small intestine were greater with urea as the supplemental N source. The level of DIP was closely associated (R2 = .89) with MNF. Postruminal digestibility of OM was greater (P < .05) for FM than for urea-supplemented diets, compensating for lower RDOM. There were no treatment effects (P > .10) on DOM. As the level of FM was increased, MIAA increased linearly (P < .01). Intestinal MIAA were similar (P > .10) for urea- and SBM-supplemented diets. Treatment effects on 56-d growth performance were evaluated using 100 medium-framed crossbred steers (231 kg). Daily weight gain (linear effect; P < .01), DM intake (linear effect; P < .10), feed efficiency (linear effect; P < .05), and diet NE (linear effect; P < .05) increased with level of FM supplementation. Daily weight gain (P < .10) and DM intake (P < .05) were greater for urea- than for SBM-supplemented diets. Using bovine tissue as the reference protein, the biological value (based on chemical score) of the intestinal chyme protein averaged 73%; methionine was first-limiting. There was a close association (R2 = .99) between methionine supply to the small intestine and observed/expected dietary NE. The metabolizable methionine requirement (MMETR, g/d) of medium-framed feedlot steers can be reliably predicted from measures of BW and ADG (MMETR = 1.565 + .0234ADG[268 - (29.4 x .0557BW(.75)ADG(1.097))/ADG] + .0896BW(.75)). There was a very close association (R2 = .89) between DIP and MNF (MNF = 13.7DIP - .66DIP(2) + 25.9). At maximal observed synthesis, DIP accounted for 76% of the MNF. A minimum of 100 g DIP/kg of total tract digestible OM was required to maximize RDOM and MNF.     

Caring for dying children: nurses' experiences

Interviews with 25 nurses in this grounded theory study show that when nurses recognized that a child's death was inevitable, they struggled with both grief distress and moral distress. Their distress occurred within the context of the nurse-patient relationship. Nurses employed a range of strategies to manage their distress. Several conditions facilitated or constrained nurses' strategies, and resulted in far-reaching implications both professionally and personally.     

Cdc34 and the F-box protein Met30 are required for degradation of the Cdk-inhibitory kinase Swe1

Ubiquitin-mediated proteolysis controls the abundance of many cell cycle regulatory proteins. Recent work in Saccharomyces cerevisiae suggests that a complex consisting of Cdc53, Skp1, and a third component known as an F-box protein (termed SCF) in combination with Cdc34 specifically targets regulatory proteins for degradation, and that substrate specificity is likely to be mediated by the F-box subunit. A screen for genetic interactions with a cdc34 mutation yielded MET30, which encodes an F-box protein. MET30 is an essential gene required for cell cycle progression and met30 mutations interact genetically with mutations in SCF components. Furthermore, physical interactions between Met30, Cdc53, Cdc34, and Skp1 in vivo provide evidence for an SCFMet30 complex. We demonstrate the involvement of Met30 in the degradation of the Cdk-inhibitory kinase Swe1. Swe1 is stabilized in met30 mutants and GST-Met30 pull-down experiments reveal that Met30 specifically binds Swe1 in vivo. Furthermore, extracts prepared from cdc34 or met30 mutants are defective in polyubiquitination of Swe1. Taken together, these data suggest that SCF-mediated proteolysis may contribute to the regulation of entry into mitosis. Our data, in combination with previously published results, also provide evidence for distinct SCF complexes in vivo and support the idea that their F-box subunits mediate SCF substrate specificity.     

PRMT 3, a type I protein arginine N-methyltransferase that differs from PRMT1 in its oligomerization, subcellular localization, substrate specificity, and regulation

Methylation is one of the many post-translational modifications that modulate protein function. Although asymmetric NG,NG-dimethylation of arginine residues in glycine-arginine-rich domains of eucaryotic proteins, catalyzed by type I protein arginine N-methyltransferases (PRMT), has been known for some time, members of this enzyme class have only recently been cloned. The first example of this type of enzyme, designated PRMT1, cloned because of its ability to interact with the mammalian TIS21 immediate-early protein, was then shown to have protein arginine methyltransferase activity. We have now isolated rat and human cDNA orthologues that encode proteins with substantial sequence similarity to PRMT1. A recombinant glutathione S-transferase (GST) fusion product of this new rat protein, named PRMT3, asymmetrically dimethylates arginine residues present both in the designed substrate GST-GAR and in substrate proteins present in hypomethylated extracts of a yeast rmt1 mutant that lacks type I arginine methyltransferase activity; PRMT3 is thus a functional type I protein arginine N-methyltransferase. However, rat PRMT1 and PRMT3 glutathione S-transferase fusion proteins have distinct enzyme specificities for substrates present in both hypomethylated rmt1 yeast extract and hypomethylated RAT1 embryo cell extract. TIS21 protein modulates the enzymatic activity of recombinant GST-PRMT1 fusion protein but not the activity of GST-PRMT3. Western blot analysis of gel filtration fractions suggests that PRMT3 is present as a monomer in RAT1 cell extracts. In contrast, PRMT1 is present in an oligomeric complex. Immunofluorescence analysis localized PRMT1 predominantly to the nucleus of RAT1 cells. In contrast, PRMT3 is predominantly cytoplasmic.     

Free fatty acid inhibition of the insulin induction of glucose-6-phosphate dehydrogenase in rat hepatocyte monolayers

Rat hepatocytes in monolayer culture were utilized to determine if the decrease in glucose-6-phosphate dehydrogenase (G6PD) activity resulting from the ingestion of fat can be mimicked by the addition of fatty acids to a chemically, hormonally defined medium. G6PD activity in cultured hepatocytes was induced several-fold by insulin. Dexamethasone or T3 did not amplify the insulin induction of G6PD. Glucose alone increased G6PD activity in cultured hepatocytes from fasted donors by nearly 500%. Insulin in combination with glucose induced G6PD an additional two-fold. The increase in G6PD activity caused by glucose was greater in hepatocytes isolated from 72 hr-fasted rats as compared to fed donor rats. Such a response was reminiscent of the “overshoot” phenomenon in which G6PD activity is induced well above the normal level by fasting-refeeding rats a high glucose diet. Addition of linoleate to the medium resulted in a significant suppression of insulin’s ability to induce G6PD, but linoleate had no effect on the induction of G6PD activity by glucose alone. A shift to the right in the insulin-response curve for the induction of G6PD also was detected for the induction of malic enzyme and acetyl-CoA carboxylase. Arachidonate (0.25 mM) was a significantly more effective inhibitor of the insulin action than linoleate was. Apparently rat hepatocytes in monolayer culture can be utilized as a model to investigate the molecular mechanism by which fatty acids inhibit the production of lipogenic enzymes. In part, this mechanism of fatty acid inhibition involves desensitization of hepatocytes to the lipogenic action of insulin.     

Deformation Bands in Ceria-Stabilized Tetragonal Zirconia/Alumina : II, Stress-Induced Aging at Room Temperature

A stress-induced aging phenomenon is observed to occur at room temperature in deformation bands introduced into a 8.5 mol% ceria-stabilized tetragonal zirconia/alumina (CeTZP/A1₂O₃) composite by flexural loading. The aging occurs with time after unloading and in laboratory air. Over a period of 100 days, the concentration of monoclinic zirconia within a deformation band increases and, in addition, the wedge-shaped deformation band grows with time. Accompanying these two changes are an increase in the tensile stress in the remaining tetragonal zirconia within the deformation band and a consequential increase in the overall compressive stress within the band. The average value of the monoclinic concentration within the deformation band is found to increase parabolically with time, suggesting the mechanism responsible for the observed aging is diffusion limited. Away from the deformation bands, no aging is observed to occur, suggesting aging is stress dependent. Although a water-vapor-mediated mechanism cannot be ruled out, it is proposed that the observed aging is in fact due to a tensile stress assisted chemical reduction of Ce⁴⁺ to Ce³⁺ whose rate is controlled by the indiffusion of oxygen vacancies driven by the tensile stress gradient. It is further proposed that the deformation band grows with time, since the region ahead of the band is under tension and hence subject to an enhanced rate of reduction     

Altered properties of the branched chain amino acid-preferring activity contribute to increased cleavages after branched chain residues by the "immunoproteasome"

The multicatalytic proteinase complex (MPC, proteasome) is assembled from 14 nonidentical protein subunits. It expresses five distinct proteolytic activities, including a chymotrypsin-like activity, cleaving after hydrophobic residues, and a branched chain amino acid-preferring component (BrAAP), cleaving preferentially after branched chain residues. Exposure of cells to interferons leads to replacement of the X, Y, and Z subunits by the LMP2, LMP7, and MECL1 subunits. This "immunoproteasome" is critical to processing of certain antigens. The enzymatic basis for enhanced antigen processing has not been determined. To gain insight into this question, we examined sites and relative rates of cleavage of bonds in denatured, reduced, carboxyamidomethylated lysozyme, a 129-amino acid protein, by MPC from bovine spleen, in which the X, Y, and Z subunits are replaced by LMP2, LMP7, and MECL1. We compared cleavages to those catalyzed by MPC from bovine pituitary, which contains only the X, Y, and Z subunits. We found marked increases in the rates and number of cleavages after branched chain residues in reduced, carboxyamidomethylated lysozyme by the spleen MPC. This was largely due to accelerated cleavages of bonds after a Phi-X-Br motif, where Phi is a hydrophobic residue, X is a small neutral or polar residue, and Br is a branched chain residue. Inhibitors with these structural properties were selective and potent inhibitors of the BrAAP activity of the spleen MPC. The above findings indicate that alterations in activity and substrate specificity of the BrAAP activity are important factors underlying the altered cleavages after hydrophobic residues associated with incorporation of interferon-inducible subunits. The potential relevance of the findings to antigen processing functions of MPC is discussed.