"We're really not ready for this": A qualitative exploration of community pharmacy personnel's perspectives on the pharmaceutical care of older people with sensory impairment.

BACKGROUND: In most developed countries there is an increasing ageing population living in the community with long-term conditions and sensory impairment (sight; hearing; dual impairment). Community pharmacy personnel are key providers of pharmaceutical care to this patient population. OBJECTIVE: This study explored community pharmacy personnel's experiences with providing pharmaceutical care for older people with sensory impairment. METHODS: Semi-structured telephone interviews were conducted with community pharmacy personnel across Scotland between 2015 and 2016. RESULTS: Thirty interviews were completed with community pharmacists (n = 17) and other pharmacy personnel (n = 13). Two overarching themes emerged: safety and communication. Interviewees reported patients' reluctance to disclose their impairment "patients are very good at hiding it" and had considerable safety concerns "it's a fear that they're going to take too much … accidentally taking the same medicine twice". Difficulties in communication were cited "no matter what you do or how you label things, leaflets and telling people, things can go wrong". Additionally, interviewees identified training needs to increase their disability awareness and to identify strategies to provide safe and reliable pharmaceutical care to this vulnerable group "We don't specifically have anything in place to deal with anyone with impairments of that kind". CONCLUSIONS: This is the first in-depth exploration of providing pharmaceutical care to older people with sensory impairment from the perspective of community pharmacy personnel. Strategies are needed to encourage older people to disclose their sensory impairment. Education and training are also needed to optimise the provision of pharmaceutical care to this vulnerable population.

Analysis of glutathione S-transferase from human liver by isoelectric focusing in a urea minigel system.

A method for the rapid analysis of isozyme subunits of glutathione transferase (GST) from human liver is described. Following purification of enzyme protein to electrophoretic homogeneity on columns of GSH-agarose, pooled transferase fractions were concentrated by ultrafiltration and subjected to further fractionation and analysis by urea-isoelectric focusing in minigels using a Hoefer Mighty Small II electrophoresis system. These methods combined with immunoblotting techniques permitted the resolution, detection, and eventual analysis of up to six different subunits of the alpha isozyme of human GST and at least three to four different forms of the pi isozyme of the transferase rapidity, accuracy, and sensitivity of the methodology may prove useful to the analysis and quantification of GST subunits in biopsies of malignant human tissue and to the development of effective chemotherapeutic regimens.

Alterations in trehalase solubility during development in the cellular slime mould Dictyostelium discoideum.

Previous studies have indicated that during development in the slime mould Dictyostelium discoideum, compartmentation of the isoenzymes of trehalase (alpha, alpha'-trehalose 1-D-glucohydrolase, (EC 3.2.1.28) occurs between the extracellular and intracellular environments. The compartmentation of trehalase between soluble and particulate cell fractions was examined in this work. The trehalase present in crude homogenates prepared during the first 12 h of development was completely soluble. Starting at about the pseudoplasmodial stage (i.e. the 14th hour of development), trehalase activity became associated with insoluble cellular material and this increased to a maximal value in homogenates from mature sorocarps, where 50% of the activity was insoluble. Spore cells accounted for only 2 to 3% of the trehalase associated with mature sorocarps, with the remaining 97% being localized in stalk cell material. Although trehalase recovered from spores was completely soluble, more than half of that from the stalk was recovered in the buffer-insoluble pellet fraction.

Trehalase from the cellular slime mold Dictyostelium discoideum: purification and characterization of the homogeneous enzyme from myxamoebae.

Trehalase (alpha-alpha'-trehalose 1-D-glucohydrolase, EC 3.2.1.28) was solubilized from myxamoebae of the cellular slime mold Dictyostelium discoideum by a freeze-thaw cycle and was subsequently purified to homogeneity using the techniques of ethanol fractionation, molecular sieve chromatography, DEAE-cellulose ion-exchange chromatography, chromatofocusing, and preparative polyacrylamide disc gel electrophoresis. The 1000-fold purified enzyme had a specific activity of about 104 units/mg, which was accompanied by a net recovery of 5 to 7% of the original activity. The purified enzyme was maximally active at pH 5.5, showed high specificity for trehalose, and exhibited a typical hyperbolic response as a function of trehalose concentration with a Km of 1.2 mM. The enzyme was maximally active at 50 degrees C and had an energy of activation of 12-13 kcal/mol. Thermal stability studies demonstrated that full enzymatic activity was recovered following a 5-min incubation of trehalase at temperatures up to 45-50 degrees C. Analysis of various compounds for inhibitory effects indicated that Tris and urea were slightly effective, reducing enzymatic activity by 28 and 6% at concentrations of 100 and 10 mM, respectively. Of five heavy metals tested, HgCl2 was the most inhibitory, reducing activity by 58% when present at a final concentration of 1.0 mM. Enzymatic activity was not affected by any adenine derivative examined (e.g., ATP, ADP, AMP, cAMP, adenosine, and adenine). The molecular weight of the native enzyme was determined by molecular sieve chromatography, pore gradient electrophoresis, and electrophoresis as a function of acrylamide concentration. All three methods yielded a value of about 10(5) +/- 5 X 10(3). Estimation of the subunit or monomer molecular weight by sodium dodecyl sulfate-gel electrophoresis indicated a value of 95-100 X 10(3). The isoelectric point as determined in 7.5% polyacrylamide gels with pH 3-10 ampholytes was 7.2-7.3. The purified enzyme adsorbed to concanavalin A-Sepharose in the presence of KCl (0.1 M) and was eluted with alpha-methylmannoside, thereby suggesting an association between trehalase and carbohydrate. In agreement with this conclusion was the observation that trehalase could be specifically stained for carbohydrate with the Alcian blue and periodic acid-Schiff's reagents following polyacrylamide disc gel electrophoresis.

Methods for the rapid purification of trehalase from Dictyostelium discoideum.

A rapid and reliable method for the preparation of homogeneous trehalase from the cellular slime mold, Dictyostelium discoideum for usage in enzyme characterization studies and trehalose assays was developed. This procedure takes advantage of the fact that trehalase activity is secreted by Dictyostelium during the course of development, the major fraction being released late in fruiting body formation. Purification of trehalase to electrophoretic homogeneity was accomplished utilizing the techniques of ultrafiltration, streptomycin sulfate precipitation, ammonium sulfate fractionation, DEAE-Sephacel chromatography and preparative disc gel electrophoresis. Analysis of the purified enzyme by analytical polyacrylamide disc gel electrophoresis demonstrated the presence of a single protein band which was stainable with Coomassie blue. Assay of trehalase activity in eluates from segments of a companion gel indicated that all of the recovered trehalase activity was associated with this band of protein. Examination of the substrate specificity of the purified enzyme indicated absolute specificity for trehalose.

Multiple forms of glucokinase from Dictyostelium discoideum.

A single form of glucokinase with an apparent Km value equal to 0.12 mM glucose was detectable in extracts prepared from aggregating cells, whereas kinetic and electrophoretic evidence indicated the presence of this form as well as a second glucose-phosphorylating enzyme with a Km value of about 0.01 mM glucose in extracts from culminating cells.

Fourth expansion and glucose perturbation of the Dictyostelium kinetic model.

The kinetic model of carbohydrate metabolism has been expanded to include: (a) the accumulation of alpha and beta-cellulose, insoluble cell-wall glycogen and mucopolysaccharide; (b) the role of RNA turnover as a source of carbon for end-product synthesis and as a buffer regulating the level of uridine nucleotides in this metabolic network; and (c) the role of purine-nucleoside phosphorylase, 5'-AMP nucleotidase, nucleosidediphosphate kinase and polynucleotide phosphorylase. One of many predictions based on this model is that cells differentiating in the presence of glucose will produce sorocarps with an abnormally high trehalose to cellulose ratio. External perturbation of either the model or of developing cells by glucose increases the levels of sorocarp trehalose and glycogen, 5-fold and 6-fold respectively. Evaluation of the experimental data and the simulation analyses have allowed several predictions to be made concerning the compartmentation of metabolites and the permeability of cells to glucose during differentiation.

Modification of the composition and structure of the yeast cell wall by culture in the presence of sulfur amino acids.

Growth of Candida utilis and Saccharomyces cerevisiae in a medium supplemented with sulfur amino acids led to synthesis and accumulation of S-adenosylmethionine, accompanied by a reduction in the cell yield, an increased sensitivity of the cell wall to snail gut enzymes (Helix pomatia), as judged by spheroplast formation, and by a modification of the chemical composition of both the intact cells and their isolated walls. Walls of supplemented cultures of C. utilis were three times as sensitive to enzymatic digestion as walls from nonsupplemented cultures. In contrast to C. utilis, walls isolated from supplemented cultures of S. cerevisiae were digested slightly more rapidly by the purified snail extract than those from nonsupplemented cultures. Chemical modifications of the cell wall are interpreted to explain the ease with which cells from sulfur amino acid-supplemented cultures are converted to spheroplasts.