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Biomedical subjects

D Lloyd

Publications and source records attributed to D Lloyd.

316 records · Page 18Linked to original sources

Routine assessment of patient progress: a multiformat, change-sensitive nurses' instrument for assessing psychotic inpatients.

A new instrument, the Routine Assessment of Patient Progress (RAPP), is a standardized scale with a unique format that allows trained nurses to incorporate both interview and observational data into a comprehensive assessment of psychiatric inpatients. The scale was developed to accurately document treatment effects, assist in treatment planning, and provide ancillary judgments of psychopathology that may affect diagnosis. The RAPP demonstrates satisfactory interrater reliability and internal consistency. The RAPP total score and its two subscales correlated highly, where expected, with an observation-based nursing scale, global measures, and an interview-based psychopathology scale. RAPP scores demonstrated sensitivity to change over time in patient functioning. It reliably discriminated among patients discharged to settings with varying degrees of independent living. The use of the RAPP over several years on a psychiatric inpatient unit has shown it to be a practical clinical tool that eases nurses' charting demands, helps create care plans, and facilitates interdisciplinary communication.

Activities of Daily Living↗

Hydrogen production by rumen holotrich protozoa: effects of oxygen and implications for metabolic control by in situ conditions.

Experiments with washed suspensions of holotrich protozoa (Isotricha spp. and Dasytricha ruminantium) showed that both organisms have an efficient O2-scavenging capability (apparent Km values 2.3 and 0.3 microM, respectively). Reversible inhibition of H2 production increased almost linearly with increasing O2 up to 1.5 microM; higher levels of O2 gave irreversible inhibition. In situ determinations of H2, CH4, O2 and CO2 in ovine rumen liquor, using a membrane inlet mass spectrometer probe, indicated that O2 was present before feeding at 1-1.5 microM and decreased to undetectable levels (less than 0.25 microM) within 25 min after feeding. A transient increase in O2 concentration after feeding occurred only in defaunated animals and resulted in suppression of CH4 and CO2 production. The presence of washed holotrich protozoa decreases the O2 sensitivity of CH4 production by suspensions of a cultured methanogenic bacterium Methanosarcina barkeri. It is concluded that holotrich protozoa play a role in ruminal O2 utilization as well as in the production of fermentation end products (especially short-chain volatile fatty acids) utilized by the ruminant and H2 utilized by methanogenic bacteria. These hydrogenosome-containing protozoa thus both control patterns of fermentation by influencing O2 levels, and are themselves regulated by the low ambient O2 concentrations they experience in the rumen.

Animals↗

Rapid methylation of cell proteins and lipids in Trypanosoma brucei.

The fate of the [methyl-14C] group of S-adenosylmethionine (AdoMet) in bloodstream forms of Trypanosoma brucei brucei, was studied. Trypanosomes were incubated with either [methyl-14C]methionine, [U-14C]methionine, S-[methyl-14C]AdoMet or [33S]methionine and incorporation into the total TCA precipitable fractions was followed. Incorporation of label into protein through methylation was estimated by comparing molar incorporation of [methyl-14C] and [U-14C]methionine to [35S]methionine. After 4-h incubation with [U-14C]methionine, [methyl-14C]methionine or [35S]methionine, cells incorporated label at mean rates of 2,880 pmol, 1,305 pmol and 296 pmol per mg total cellular protein, respectively. Cells incubated with [U-14C] or [methyl-14C]methionine in the presence of cycloheximide (50 micrograms/ml) for four hours incorporated label eight- and twofold more rapidly, respectively, than cells incubated with [35S]methionine and cycloheximide. [Methyl-14C] and [U-14C]methionine incorporation were > 85% decreased by co-incubation with unlabeled AdoMet (1 mM). The level of protein methylation remaining after 4-h treatment with cycloheximide was also inhibited with unlabeled AdoMet. The acid precipitable label from [U-14C]methionine incorporation was not appreciably hydrolyzed by DNAse or RNAse treatment but was 95% solubilized by proteinase K. [U-14C]methionine incorporated into the TCA precipitable fraction was susceptible to alkaline borate treatment, indicating that much of this label (55%) was incorporated as carboxymethyl groups. The rate of total lipid methylation was found to be 1.5 times that of protein methylation by incubating cells with [U-14C]methionine for six hours and differential extraction of the TCA lysate. These studies show T. b. brucei maintains rapid lipid and protein methylation, confirming previous studies demonstrating rapid conversion of methionine to AdoMet and subsequent production of post-methylation products of AdoMet in African trypanosomes.

Animals↗

Effects of carboxylmethylation and polyamine synthesis inhibitors on methylation of Trypanosoma brucei cellular proteins and lipids.

The fate of methionine in eukaryotic cells is divided between protein synthesis and the branched pathway encompassing polyamine synthesis, methylation of proteins and lipids, and transsulphuration reactions. Aside from protein synthesis, the first step to all other uses of methionine is conversion to S-adenosylmethionine. Blockade of polyamine synthesis in African trypanosomes by the ornithine decarboxylase inhibitor DL-alpha-difluoromethylornithine (Ornidyl, DFMO) the AdoMet decarboxylase inhibitor 5'-[[(Z)-4-amino-2-butenyl]-methylamino]-5'-deoxyadenosine or the protein methylase inhibitor sinefungin induces dramatic increases in intracellular AdoMet. In a previous study, distribution and pool sizes of [35S] or [U-14C]methionine were followed in bloodform trypanosomes as incorporation into the total TCA precipitable fractions. In the present study, the effects of pretreatment with DFMO (1 mM), MDL 73811 (1 microM) and sinefugin (2 nM) on [35S] and [U-14C]methionine incorporation were studied in blood forms. DFMO or MDL 73811 pretreatment increased protein methylation 1.5-fold through incorporation of [U14C]methionine, while sinefungin caused a 40% reduction of incorporation. The increases in incorporation of [U-14C]methionine due to DFMO and MDL 73811 were reduced 40% to 70% by including cold AdoMet (1 mM) in the incubation medium, an indication of AdoMet transport by bloodform trypanosomes and the utilization of [U-14C]methionine as AdoMet. Exogenous AdoMet had no effect on [35S]methionine incorporation. The agents studied are curative for African trypanosomiasis infections, either clinically (DFMO) or in model infections (MDL 73811, sinefungin) and thus highlight interference with AdoMet metabolism and methylation reactions as biochemical consequences of these agents.

Adenosine↗

Oxygen uptake and antioxidant responses of the free-living diplomonad Hexamita sp.

The free-living anaerobic flagellate Hexamita sp. was observed to actively consume O2 with a K(m) O2 of 13 microM. Oxygen consumption increased linearly with O2 tension up to a threshold level of 100 microM, above which it was inhibited. Oxygen uptake was supported by a number of substrates but probably not coupled to energy conservation as cytochromes could not be detected spectro-photometrically. In addition, inhibitors specific for respiratory chain components did not significantly affect O2 uptake. Respiration was however, partially inhibited by flavoprotein and iron-sulfur protein inhibitors. NAD(P)H supported O2 consumption was measured in both particulate and soluble fractions; this activity was partially inhibited by quinacrine. A chemosensory response was observed in cells exposed to air, however no response was observed in the presence of superoxide dismutase plus catalase. Catalase and nonspecific peroxidase activity could not be detected, but superoxide dismutase plus catalase. Catalase and nonspecific peroxidase activity could not be detected, but superoxide dismutase activity was present. Superoxide dismutase was sensitive to NaN3, and H2O2 but not KCN, suggesting a Fe prosthetic group. Flow cytometric analysis revealed that thiol levels in live cells were depleted in the presence of t-butyl H2O2. The observed NADPH-driven glutathione reductase activity is believed to recycle oxidized thiols in order to re-establish reduced thiol levels in the cell. The corresponding thiol cycling enzyme glutathione peroxidase could not be detected. The ability to withstand high O2 tensions (100 microM) would enable Hexamita to spend short periods in a wider range of habitats. Prolonged exposure to O2 tensions higher than 100 microM leads to irreversible damage and cell death.

Anaerobiosis↗

Encystation in Acanthamoeba castellanii: development of biocide resistance.

Since the early 1960s, axenic culture and the development of procedures for the induction of encystation have made Acanthamoeba spp. superb experimental systems for studies of cell biology and differentiation. More recently, since their roles as human pathogens causing keratitis and encephalitis have become widely recognized, it has become urgent to understand the parameters that determine differentiation, as cysts are much more resistant to biocides than are the trophozoites. Viability of trophozoites of the soil amoeba Acanthamoeba castellanii (Neff), is conveniently measured by its ability to form plaques on a lawn of Escherichia coli. Use of confocal laser scanning microscopy with Calcofluor white, Congo Red or the anionic oxonol dye, DiBAC4(3) or flow cytometry with propidium iodide diacetate and fluorescein or oxonol provides more rapid assessment. For cysts, the plaque method is still the best, because dye exclusion does not necessarily indicate viability and therefore the plate count method has been used to study the sequence of development of biocide resistance during the differentiation process. After two hours, resistance to HCl was apparent. Polyhexamethylene biguanide, benzalkonium chloride, propamidine isethionate, pentamidine isethionate, dibromopropamine isethionate, and H2O2 and moist heat, all lost effectiveness at between 14 and 24 h after trophozoites were inoculated into encystation media. Chlorhexidine diacetate resistance was observed at between 24 and 36 h. The molecular biology and biochemistry of the modifications that underlie these changes are now being investigated.

Acanthamoeba↗

The effect of patient ethnicity on prescribing rates.

The issuing of a prescription is central to any doctor-patient interaction. Prescribing variation exists and remains largely unexplained. There is little documented evidence of the effect of patient ethnicity on prescribing patterns. We carried out a secondary analysis of data from the General Household Surveys to examine the association between being given a prescription and patient ethnicity. After modelling, we found that Pakistanis and Indians were significantly more likely to receive a prescription from their general practitioner at a consultation compared to white and West Indian ethnic groups. In addition, consultation rate explained the different prescribing rates among women and men in the white group only. This study raises further questions of the underlying reasons causing these differences which need answering.

Adolescent↗

Primary care: nurse-led telephone triage and advice out-of-hours.

In this article the authors report on the evaluation of an out-of-hours telephone triage and advice service in general practice. A computer-based decision support tool was used to guide nurses in the assessment of patients and outcome for calls. Data from the computer system were analysed for all calls over a six-month period. The researchers found that the nurses were able to handle just over half the calls received by giving advice alone. Overall, the service appeared to be remarkably consistent in the decisions taken by nurses, and training and organisational issues to be considered in the future development of the service were identified.

Adolescent↗