Medicine burden experiences of people living with HIV and association with stigma.

The medicine burden of people living with HIV (PLWH) is unknown. Between 2018 and 2020, participants completed a survey comprising outcome measures for medicine burden (LMQ-3) and stigma experiences (SSCI-8). Participants were HIV+ adults (≥18 years), using antiretrovirals (ARV) with or without non-ARV medicines, recruited via two outpatient clinics in southeast England and online via HIV charities across the UK. Spearman's correlations between medicine burden levels and stigma scores were calculated. Participants were mostly males (72%, 101/141) of mean (SD) age 48.6 (±12.31) years. Total number of medicines ranged from 1-20. High medicine burden was self-reported by 21.3% (30) and was associated with polypharmacy (≥ 5 medicines) (101.52 Vs 85.08, p = 0.006); multiple doses versus once daily regimes (109.31 Vs 85.65, p = 0.001); unemployment (98.23 Vs 84.46, p = 0.004); and ethnicity (97 Vs 86.85, p = 0.041 for non-White versus White participants). A correlation between medicine burden and stigma was observed (r = 0.576, p < 0.001). The LMQ-3 demonstrated adequate construct validity and reliability (domain loadings ranging 0.617-0.933 and Cronbach's α of 0.714-0.932). Assessment of medicine burden and psychosocial stigma in PLWH could enable identification of those needing additional support in future research and practice.

Public attitudes towards community pharmacy attributes and preferences for methods for promotion of public health services.

OBJECTIVE: To identify attitudes towards pharmacy characteristics and promotional methods for selected pharmacy public health services (lifestyle advice and screening for cardiovascular risk factors) among different sectors of the general public. STUDY DESIGN: Cross-sectional survey, using a previously validated questionnaire. METHODS: Three survey methods were used, across 15 areas of England, to maximize diversity: face-to-face; telephone; and self-completion of paper questionnaires. Responses to closed questions regarding characteristics and promotion were quantified and differences among sub-groups explored by univariate and multivariate analysis. RESULTS: In total, 2661 responses were available for analysis: 2047 face-to-face; 301 telephone; and 313 paper. There were strong preferences for a pharmacy near to home or doctor's surgery and for long opening hours, particularly among employed people and non-whites. Fifty percent preferred not to use a pharmacy in a supermarket, particularly older people, the retired, those of lower education and frequent pharmacy users. Personal recommendation by health professionals or family/friends was reported as most likely to encourage uptake of pharmacy public health services, with older people and males being less likely and frequent pharmacy users more likely to perceive any promotional method as influential. Posters/leaflets were preferred over mass-media methods, with fewer than 30% perceiving the latter as potentially influential. CONCLUSION: Pharmacists, pharmacy companies and service commissioners should use promotional methods favoured by potential users of pharmacy public health services and be aware of differences in attitudes when trying to reach specific population sub-groups. For personal recommendation to be successful, good inter-professional working and a pro-active approach to existing customers are needed.

Towards improving service delivery in screening and intervention services in community pharmacies: a case study of an alcohol IBA service.

BACKGROUND: Previous studies have demonstrated positive outcomes from a range of pharmacy public health services, but barriers to delivery remain. This paper explores the processes of delivering an alcohol screening and intervention service, with a view to improving service delivery. METHODS: A mixed-methods, multi-perspective approach was used, comprising in-pharmacy observations and recording of service provision, follow-up interviews with service users and interactive feedback sessions with service providers. RESULTS: Observations and recordings indicate that staff missed opportunities to offer the service and that both availability and delivery of the service were inconsistent, partly owing to unavailability of trained staff and service restrictions. Most service users gave positive accounts of the service and considered pharmacies to be appropriate places for this service. Respondents also described positive impacts, ranging from thinking more about alcohol consumption generally to substantial reductions in consumption. Key facilitators to service provision included building staff confidence and service champions. Barriers included commissioning issues and staff perception of alcohol as a sensitive topic. CONCLUSIONS: Findings support expansion of pharmacies' role in delivering public health services and highlight benefits of providing feedback to pharmacy staff on their service provision as a possible avenue for service improvement.

General public's views on pharmacy public health services: current situation and opportunities in the future.

OBJECTIVES: To explore the experience of and willingness to use seven pharmacy public health services related to cardiovascular risk among the general public in England. STUDY DESIGN: Mixed-methods study. METHODS: A mixed-methods study, involving a cross-sectional survey using multiple distribution methods followed by a focus group discussion (FGD) with a sample of survey respondents. RESULTS: From 3596 approachable individuals, 908 questionnaires were completed (response rate 25.3%). Few respondents (2.1-12.7%) had experienced any of the seven pharmacy public health services. About 40% stated they would be willing to use health check services, fewer (9.3-26.3%) were willing to use advisory services. More females, frequent pharmacy users and those in good health were willing to use services in general (P < 0.05). Smokers, overweight individuals and those with alcohol-related problems were most willing to use specific advisory services supporting their problems (P < 0.05). FGD identified barriers to service use; for example, frequent staff changes, seeing pharmacist as medicines suppliers and concerns about competence for these services. CONCLUSION: The general public are receptive to pharmacy public health services. Pharmacists must consider barriers if uptake of services is to increase.

Involving the public and other stakeholders in development and evaluation of a community pharmacy alcohol screening and brief advice service.

OBJECTIVES: To explore the views of community pharmacy staff, the general public and other stakeholders towards pharmacy-based alcohol screening and advice services. To involve all relevant stakeholders in designing an acceptable and feasible pharmacy-based alcohol screening and advice service. To evaluate a pilot service from the user perspective. STUDY DESIGN: Mixed methods study involving a range of populations, designed to explore multiple perspectives and enable triangulation of results, to develop an optimal service design, prior to service commissioning. METHODS: Telephone interviews were conducted with relevant stakeholders and a street survey undertaken with the public to explore views on the desirability and feasibility of pharmacy-based alcohol services. Following this, a stakeholder working group was held, involving a nominal group technique, to develop and refine the service design. Finally a pilot service was evaluated from the user perspective through telephone interviews and direct observations by a trained researcher. RESULTS: All stakeholder groups (pharmacy staff, public, commissioners, alcohol treatment service staff) viewed pharmacy-based alcohol screening services as acceptable and feasible with the potential for integration and/or combination with existing public health services. Privacy was the main concern of the public, but 80% were comfortable discussing alcohol in a pharmacy. These views were not influenced by drinking status age or gender, but people recruited in areas of high deprivation were more likely to accept a pro-active approach or alcohol-related advice from a pharmacist than those from areas of low deprivation. Stakeholder groups were in agreement on the acceptability of a pharmacy screening service, but alcohol treatment service staff viewed direct referral to alcohol support services less beneficial than other stakeholders. Posters in pharmacies and GP surgeries were viewed as most likely to encourage uptake of screening. Involvement of non-pharmacist pharmacy staff was seen as essential. The working group considered accessibility of pharmacies as the key facilitator for alcohol services, but agreed that an optimal service must ensure that poor pharmacy environment did not create a potential barrier, that clear information about the service's availability was necessary. Plus good use of quiet areas. Use of AUDIT-C as a prescreening tool by pharmacy staff, followed when appropriate by completion of full AUDIT by the pharmacist in a private room/quiet area was agreed as optimal to ensure accessibility plus privacy. Direct referral was viewed as desirable. Five pharmacies piloted this service for two months and recruited 164 people for alcohol screening, of whom 113 were low risk (AUDIT score 0-7), 24 increasing risk (8-19) and 28 high risk/possibly dependent drinkers (20 or above). Observations showed that pharmacy support staff were involved in proactively approaching customers, that 20 of the 72 customers observed (28%) during two hours in each pharmacy were invited for screening and that 14 (19%) accepted screening. Promotion of the service was variable dependent on company policies, but was shown to have a positive effect, as two of the ten service users interviewed requested screening. The environment was judged suitable for alcohol services in all pharmacies, but some quiet areas were not audibly discrete. Ten service users interviewed all considered the experience positive and all would recommend the service, but most wanted the service to be delivered in a private area. CONCLUSION: The methodology enabled the development of pharmacy-based alcohol screening to be assessed for acceptability and feasibility from multiple perspectives, prior to full service commissioning. Results suggest that the pharmacy environment and concerns about privacy need to be recognized as potential barriers to service delivery. Good promotion is required to maximize service uptake and pharmacy staff need to be involved in both this and in service delivery.

Evaluation of patient reporting of adverse drug reactions to the UK 'Yellow Card Scheme': literature review, descriptive and qualitative analyses, and questionnaire surveys.

BACKGROUND: The monitoring of adverse drug reactions (ADRs) through pharmacovigilance is vital to patient safety. Spontaneous reporting of ADRs is one method of pharmacovigilance, and in the UK this is undertaken through the Yellow Card Scheme (YCS). Yellow Card reports are submitted to the Medicines and Healthcare products Regulatory Agency (MHRA) by post, telephone or via the internet. The MHRA electronically records and reviews information submitted so that important safety issues can be detected. While previous studies have shown differences between patient and health-care professional (HCP) reports for the types of drugs and reactions reported, relatively little is known about the pharmacovigilance impact of patient reports. There have also been few studies on the views and experiences of patients/consumers on the reporting of suspected ADRs. OBJECTIVES: To evaluate the pharmacovigilance impact of patient reporting of ADRs by analysing reports of suspected ADRs from the UK YCS and comparing reports from patients and HCPs. To elicit the views and experiences of patients and the public about patient reporting of ADRs. DESIGN: (1) Literature review and survey of international experiences of consumer reporting of ADRs; (2) descriptive analysis of Yellow Card reports; (3) signal generation analysis of Yellow Card reports; (4) qualitative analysis of Yellow Card reports; (5) questionnaire survey of patients reporting on Yellow Cards; (6) qualitative analysis of telephone interviews with patient reporters to the scheme; (7) qualitative analysis of focus groups and usability testing of the patient YCS; and (8) national omnibus telephone survey of public awareness of the YCS. PARTICIPANTS: Patients (n = 5180) and HCPs (n = 20,949) submitting Yellow Card reports from October 2005 to September 2007. Respondents to questionnaire survey (n = 1362). Participants at focus groups and usability testing sessions (n = 40). National omnibus telephone survey (n = 2028). SETTING: The literature review included studies in English from across the world. All other components included populations from the UK; the omnibus survey was restricted to Great Britain. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Characteristics of patient reports: types of drug and suspected ADR reported; seriousness of reports; and content of reports. The relative contributions of patient reports and of HCP reports to signal generation. Views and experiences of patient reporters. Views of members of the public about the YCS, including user-friendliness and usability of different ways of patient reporting. Public awareness of the YCS. Suggestions for improving patient reporting to the YCS. RESULTS: Compared with HCPs, patient reports to the YCS contained a higher median number of suspected ADRs per report, and described reactions in more detail. The proportions of reports categorised as 'serious' were similar; the patterns of drugs and reactions reported differed. Patient reports were richer in their descriptions of reactions than those from HCPs, and more often noted the effects of ADRs on patients' lives. Combining patient and HCP reports generated more potential signals than HCP reports alone; some potential signals in the 'HCP-only' data set were lost when combined with patient reports, but fewer than those gained; the addition of patient reports to HCP reports identified 47 new 'serious' reactions not previously included in 'Summaries of Product Characteristics'. Most patient reporters found it fairly easy to make reports, although improvements to the scheme were suggested, including greater publicity and the redesign of web- and paper-based reporting systems. Among members of the public, 8.5% were aware of the YCS in 2009. CONCLUSIONS: Patient reporting of suspected ADRs has the potential to add value to pharmacovigilance by reporting types of drugs and reactions different from those reported by HCPs; generating new potential signals; and describing suspected ADRs in enough detail to provide useful information on likely causality and impact on patients' lives. These findings suggest that further promotion of patient reporting to the YCS is justified, along with improvements to existing reporting systems. In order of priority, future work should include further investigation of (1) the pharmacovigilance impact of patient reporting in a longer-term study; (2) the optimum approach to signal generation analysis of patient and HCP reports; (3) the burden of ADRs in terms of impact on patients' lives; (4) the knowledge and attitudes of HCPs towards patient reporting of ADRs; (5) the value of using patient reports of ADRs to help other patients and HCPs who are seeking information on patient experiences of ADRs; and (6) the impact of increasing publicity and/or enhancements to reporting systems on the numbers and types of Yellow Card reports from patients. FUNDING: The National Institute for Health Research Health Technology Assessment programme.

Hospital pharmacists' activities and attitudes regarding the Thai safety monitoring program for new drugs.

PURPOSE: To determine the knowledge, attitudes and practices of Thai hospital pharmacists concerning the safety monitoring programme and reporting of adverse drug reactions to the Thai Food and Drug Administration. METHODS: Postal questionnaires distributed to 414 Thai hospitals, including all larger hospitals and a proportion of small community hospitals, for completion by the pharmacist responsible for ADR reporting to the Thai FDA. RESULTS: The response rate was 63.0% (n = 262) with most respondents (190; 72.8%) being directly responsible for ADR reporting. Most hospitals (251; 96.5%) designated at least one pharmacist as responsible for ADR reporting, but only 164 (62.8%) respondents were aware of the SMP. Only 15 respondents out of the 56 working in hospitals using new drugs had established reporting via the SMP. Uncertainty in ascribing causality, lack of co-operation between health professionals and lack of staff were identified as barriers to reporting. Most respondents agreed that enhanced co-operation and improvements needed in hospital systems were required. Less than half agreed that the reporting system was simple and the majority agreed that changes were required to the SMP reporting mechanisms. CONCLUSIONS: Thai hospital pharmacists have positive attitudes towards reporting of ADRs, but not all are aware of the SMP for new drugs. Few hospitals use new drugs and even less are involved in active reporting of suspected ADRs via the SMP. To increase reporting rates, potential difficulties with the organization of reporting via the SMP and the potential role of hospital pharmacists in ADR identification should be investigated further.

Adverse drug reaction monitoring: comparing doctor and patient reporting for new drugs.

PURPOSE: To compare the frequency of adverse drug reactions (ADRs) recorded by doctors with perceived ADRs reported by patients during hospitalization and post-discharge and to compare both to formal reporting in Thailand. SETTING: Tertiary teaching hospital, North-East Thailand. METHODS: Over a 6-month period doctors were requested to record ADRs to 13 recently marketed index drugs, occurring during hospitalization and after discharge, using specific forms. Patient perceived ADRs were detected by daily pharmacist interviews during hospitalization and a symptom checklist questionnaire after discharge. RESULTS: In total, 203 in-patients were prescribed an index drug and interviewed. Doctors recorded 13 ADRs in these patients, resulting in seven drugs being stopped. In-patients reported 15 further symptoms, but 12 were assessed as unlikely to be an ADR. Of 160 patients who continued the index drug after discharge, 152 accepted questionnaires and 88 (58%) were returned. Patients identified 249 symptoms, of which the pharmacist assessed 190 (76%) as probably or possibly ADRs, using a previously developed method. Only 26 symptoms were noted in out-patient department (OPD) records, although most patients claimed to have informed doctors about them. Doctors recorded only five symptoms on ADR forms, all associated with drug discontinuation. During the study period, four reports were received by the Thai FDA on these 13 drugs, none from this hospital. CONCLUSIONS: Forms encouraging doctors to identify ADRs may be an effective method of increasing recording, particularly in the in-patient setting. A combination of methods including patient reporting may enable pharmacists to increase formal ADR reporting.

Evaluation of medication reviews conducted by community pharmacists: a quantitative analysis of documented issues and recommendations.

UNLABELLED: What is already known about this subject? There is conflicting evidence concerning the potential benefits of pharmacist-led medication review. Little work has been published on the completeness of medication reviews provided by community pharmacists. What this study adds. The 60 community pharmacists taking part in a large randomized controlled trial showed considerable variation in the completeness of the reviews they recorded for intervention patients. Overall, pharmacists recorded only a minority of the potential issues present in these patients. The frequency with which pharmacists recorded issues was not related to key characteristics or to the number of reviews completed. AIMS: To describe issues noted and recommendations made by community pharmacists during reviews of medicines and lifestyle relating to coronary heart disease (CHD), and to identify and quantify missed opportunities for making further recommendations and assess any relationships with demographic characteristics of the pharmacists providing the reviews. METHODS: All issues and recommendations noted by 60 community pharmacists during patient consultations were classified and quantified. Two independent reviewers studied a subsample of cases from every participating pharmacist and identified and classified potential issues from the available data. The findings of the pharmacists and the reviewers were compared. Relevant pharmacist characteristics were obtained from questionnaire data to determine relationships to the proportion of potential issues noted. RESULTS: A total of 2228 issues and 2337 recommendations were noted by the pharmacists in the 738 patients seen, a median of three per patient (interquartile range 2-4). The majority of the recommendations made (1719; 74%) related to CHD. In the subsample of 169 patients (23% of the total), the reviewers identified 1539 potential issues, of which pharmacists identified an average of 33.8% (95% confidence interval 30.1, 36.4). No relationship was found between the proportion of issues noted and potentially relevant factors such as pharmacists' characteristics and their experience of doing reviews. CONCLUSIONS: The majority of issues and recommendations noted by pharmacists related to CHD, although pharmacists recorded only a minority of the issues identified by reviewers. Variation between pharmacists in the completeness of reviews was not explained by review or other relevant experience.

Patient reporting of potential adverse drug reactions: a methodological study.

AIMS: To develop a systematic generic method of enabling patients to report symptoms which they believe to be due to a particular prescribed drug. METHODS: A piloted body system-based questionnaire was distributed to patients registered with 79 medical practices in Grampian prescribed one of nine recently marketed 'black triangle' drugs. These comprised four antidepressants, three antiepileptics and two analgesics. This requested respondents to identify any symptoms experienced over the previous year which they thought could be due to the 'black triangle' drug they had used. A sample of medical records was examined to compare symptoms recorded with those reported by patients. A classification system was developed for the study to enable the assessment of symptoms reported for their potential relationship to patients' drug therapy. All symptoms reported were classified, taking into account information provided by patients on their concomitant drugs and diseases. A specialist pharmacist independently re-classified a sample of the symptoms to validate the process. RESULTS: A 36.3% response rate was obtained (837/2307) with 742 respondents (88.6%) reporting at least one symptom. The median per patient was 6.0 (range 0--71), with almost half (406, 48.5%) reporting fewer than five symptoms. Most symptoms (71.0%) were classified as being probably or possibly related to the drugs studied. Agreement between researcher and specialist on the classification of 75.3% of 716 symptoms was obtained (Kappa=0.563). Responses from patients prescribed antidepressant drugs were more likely to include symptoms potentially caused by these drugs (74.5% of all symptoms reported) than those from patients prescribed analgesics (67.4%) or antiepileptics (65.1%, chi2 = 23.858, d.f. = 2, P < 0.001). Patients reporting large numbers of symptoms were more likely to report some which were classed as unlikely to be an ADR or unattributable (chi2 = 80.587, d.f. = 3, P < 0.001). Of the 742 reporting symptoms in questionnaires, 402 (54.2%) claimed to have reported some or all of these to their doctor. Only 162 (22.6%) of 716 patient-reported symptoms were documented in the primary care medical records of 103 patients prescribed tramadol or venlafaxine. CONCLUSIONS: Respondents were clearly willing to report symptoms, the majority of which were classed as possibly/probably related to the drugs studied. The results suggest that patients do not report all symptoms they suspect to be ADRs to their GP and that GPs do not record all symptoms which may be reported to them. The method could help to identify problems which patients perceive as being related to their drug therapy and contribute to increased ADR reporting.

Pharmacist-led medication review in patients over 65: a randomized, controlled trial in primary care.

BACKGROUND: regular medication review has been recommended for those over 75 and those on multiple drug therapy. Pharmacists are a potential source of assistance in reviewing medication. Evidence of the benefits of this process is needed. OBJECTIVE: to study the effect of medication review led by a pharmacist on resolution of pharmaceutical care issues, medicine costs, use of health and social services and health-related quality of life. DESIGN: randomized, controlled trial. SETTING: general medical practices in the Grampian region of Scotland. SUBJECTS: patients aged at least 65 years, with at least two chronic disease states who were taking at least four prescribed medicines regularly. METHODS: pharmacists reviewed the drug therapy of 332 patients, using information obtained from the practice computer, medical records and patient interviews. In 168 patients, a pharmaceutical care plan was then drawn up and implemented. The 164 control patients continued to receive normal care. All outcome measures were assessed at baseline and after 3 months. RESULTS: all patients had at least two pharmaceutical care issues at baseline. Half of these were identified from the prescription record, the rest from notes and patient interview. Of all the issues, 21% were resolved by information found in notes and 8.5% by patient interview. General practitioners agreed with 96% of all care issues documented on the care plans in the intervention group. At the time of follow-up, 70% of the remaining care issues had been resolved in the intervention group, while only 14% had been resolved in the control group. There were no changes in medicine costs or health-related quality of life in either group. There were small increases in contacts with health-care professionals and slightly fewer hospital admissions among the intervention group than the control group. CONCLUSIONS: pharmacist-led medication review has the capacity to identify and resolve pharmaceutical care issues and may have some impact on the use of other health services.

Drug utilization evaluation of nonprescription H2-receptor antagonists and alginate-containing preparations for dyspepsia.

AIMS: To evaluate the use, efficacy and adverse effects of nonprescription H2-receptor antagonists and alginate-containing preparations obtained from community pharmacies. METHODS: Questionnaires were distributed to customers from 39 pharmacies in Scotland and Wales. RESULTS: Of 767 customers recruited, 608 (79.3%) returned an initial questionnaire and 472 (61.5%) customers a second questionnaire. The vast majority of respondents (424, 69.7%) had suffered their symptoms on three or more occasions and 369 (60.7%) had previously tried medicines to relieve their symptoms. Referrals to a doctor were less frequent than recommended in guidelines and few of those who were referred actually saw a doctor. Over a quarter of those returning the second questionnaire claimed to be taking more than one product simultaneously for symptom control. Eight customers who were taking prescribed ulcer-healing drugs obtained H2-receptor antagonists. The majority of respondents (355/472, 75. 2%) obtained some or complete symptom relief using the product obtained and 369/472 (78.2%) were completely satisfied with their product. H2-receptor antagonists were more likely to produce complete relief of symptoms than alginate-containing preparations (P < 0.05). Only 14 respondents (3.0%) reported side-effects from the product used which were mostly gastro-intestinal. CONCLUSIONS: The study demonstrated that drug utilization studies are feasible to carry out in a community pharmacy setting. While the results support published evidence of the efficacy and minimal toxicity of these products, they also highlight the possibility of H2-receptor antagonists being used outwith their licenced indications.

Withdrawal of peripheral vasodilators in primary care.

AIMS: To investigate the use of peripheral vasodilators in selected practices in primary care and to observe the effect of withdrawing therapy in a sample of patients. METHODS: Patients receiving repeat prescriptions for peripheral vasodilators, identified from two practices in Grampian, were interviewed prior to and following withdrawal of therapy. RESULTS: Of the 35 patients, review of the continued need for therapy was documented in the medical notes of only one patient. Treatment was successfully withdrawn from 17 patients, generating considerable savings. CONCLUSIONS: Review of peripheral vasodilators merits further attention in larger numbers of patients.

Thrombolytics--a drug utilization review in a district general hospital.

BACKGROUND: Retrospective drug utilization review (DUR) can be of benefit in determining the extent to which recommendations derived from clinical trials are carried out in practice. OBJECTIVE: To determine whether thrombolysis was given in accordance with hospital guidelines. METHOD: A retrospective DUR of the treatment of patients with a diagnosis of myocardial infarction (MI) admitted to Borders General Hospital during April-December 1993 was carried out. RESULTS: Forty-eight per cent of these patients received thrombolysis, representing 52% of all patients with a definite diagnosis of MI. The most frequent reason for thrombolysis not being given was atypical presentation, particularly ECG changes not in accordance with those described in the hospital guidelines for diagnosis of MI. Contraindications and late presentations were very few in number. Appropriate reasons were identified for all patients who did not receive thrombolytic therapy. Mortality both at discharge and at 6 months was slightly greater in these patients than in treated patients. CONCLUSION: Thombolytic drugs were used in line with the hospital guidelines in most cases, although there was no identifiable reason for the use of alteplase instead of streptokinase in two cases.

Adherence to the Grampian joint drug formulary in general practice.

A joint drug formulary covering both hospital and general practice prescribing has been in existence in Grampian since 1990. The aims of this study were to measure adherence to this formulary in general practice and to identify who had initiated non-formulary drugs. The drug therapy of elderly patients on presentation to hospital was compared to the formulary. Of the 449 drugs prescribed 377 (84%) were recommended in the formulary. The main areas of non-formulary prescribing were antidepressants, hypnotics, nitrates and diuretics, with some non-formulary drugs being initiated by hospital prescribers. The method described was found to be extremely simple and had the advantage that it reflected actual prescribing at the time of the study.

Sleep quality and the use of benzodiazepine hypnotics in general practice.

The aim of the study was to determine the usage patterns of benzodiazepine hypnotics, the use of other ways of improving sleep and general sleep characteristics in patients prescribed these drugs on a regular basis and in a control group of age- and sex-matched patients not receiving hypnotics. Patients receiving regular benzodiazepine hypnotic prescriptions and a control group matched for age and sex were identified from a general practice computer. They were interviewed in their own homes about their habits relating to sleep, hypnotic use and attitudes to sleep and hypnotics. None of the recommended methods of improving sleep differed significantly in frequency between the two groups or had an effect on sleep parameters. Only daytime napping was found to be slightly more frequent in hypnotic users, but had no apparent effect on self-estimates of sleep duration or latency. The hypnotic users claimed a longer sleep latency and a shorter sleep duration than non-users. More patients who took hypnotics also perceived them to be beneficial and more convenient than other ways of promoting sleep. The recommendation of sleep-promoting methods such as caffeine and alcohol avoidance and regular exercise may not be helpful in patients attempting to avoid hypnotics, as many patients already claim to use such methods.

Monoclonal antibody recognition and function of a DnaK (HSP70) epitope found in gram-negative bacteria.

The isolation and characterization of a monoclonal antibody (MAb 2G5) specific for the bacterial DnaK (HSP70) protein is described. The 2G5 MAb was initially selected because of its ability to bind to DnaK under denaturing conditions. Isotype analyses indicated that 2G5 was an immunoglobulin G2a. Dose-response Western blot (immunoblot) experiments with purified but unconcentrated 2G5 permitted detection of 10 ng of pure DnaK protein. The DnaK epitope was determined by Western blot analysis of a series of truncated DnaK fragments overproduced in Escherichia coli using 5' and 3' dnaK-deleted expression plasmids. The epitope mapped to a 22-amino-acid region spanning DnaK residues 288 and 310. Phylogenetic distribution of the epitope was examined by Western blot analysis of a wide variety of bacterial species and indicated that the epitope was uniquely present in gram-negative organisms. The proximity of the epitope to the presumed DnaK ATP-binding pocket suggested that MAb binding might inhibit DnaK ATPase activity. In vitro analysis supported this prediction and demonstrated that MAb-mediated inhibition of ATPase activity was antibody specific and occurred at stoichiometric molar ratios of MAb to DnaK. Possible mechanisms to explain the ability of the 2G5 MAb to inhibit DnaK activity are discussed.

Nucleotide sequence analysis and heterologous expression of the Erysipelothrix rhusiopathiae dnaJ gene.

DNA sequence analysis of chromosomal DNA from the Gram-positive facultative intracellular pathogen, Erysipelothrix rhusiopathiae has identified a dnaJ heat shock gene homolog. A 1109-bp open reading frame encoding dnaJ is located immediately 3' to the E. rhusiopathiae dnaK gene. The deduced DnaJ amino acid sequence exhibits the modular structure of other members of the DnaJ protein class including a glycine-rich region and the repeating consensus sequence CXXCXGXGX. Heterologous expression of the dnaJ sequence in Escherichia coli resulted in accumulation of a unique 38.9-kDa protein with an isoelectric point of 8.0. Deletion analysis of the dnaJ gene was used to confirm that the overproduced protein was encoded by the dnaJ sequence.

Cloning, heterologous expression, and characterization of the Erysipelothrix rhusiopathiae DnaK protein.

The dnaK (hsp70) gene from the facultative intracellular pathogen Erysipelothrix rhusiopathiae was cloned by heterologous DNA hybridization of a genomic library using the Escherichia coli dnaK gene as a probe. A 3.2-kb fragment which encoded an 1,800-bp open reading frame was recovered. The deduced amino acid sequence of this open reading frame shares 56% identity with the E. coli DnaK protein. Expression of the encoded protein in E. coli by using the phage T7 promoter/polymerase system resulted in accumulation of a unique 65-kDa protein. Western blot (immunoblot) analysis of extracts from a recombinant E. coli strain using anti-E. coli DnaK polyclonal antibodies confirmed that the cloned gene encodes a DnaK homolog. The recombinant E. rhusiopathiae DnaK protein was purified to 80% homogeneity by ATP affinity chromatography. The purified material hydrolyzed ATP with a specific activity of 100 nmol min-1 mg of protein-1. Analysis of total protein extracts from E. rhusiopathiae indicates that DnaK is a highly expressed protein in this organism.