Culture, diversity and inclusion: a survey of British Hip Society members.

AIM: To explore the perceptions and experiences of members of the British Hip Society (BHS) as they relate to culture, diversity and inclusion in the professional sphere. METHOD: BHS members participated in an anonymised online survey in 2021. Quantitative and qualitative data were collected on demographics, professional experiences and perceptions of workplace culture. Members provided suggestions for improving working culture and supporting inclusivity. RESULTS: A 45% response rate (n=217) was achieved. Most respondents were male consultant surgeons, of white ethnicity. Almost a quarter of respondents reported experiencing barriers to career progression within the hip subspecialty. Experience of barriers was more common among women and those of non-white ethnicity. Several members experienced an elitist, exclusive culture in the BHS which is closed to outsiders. Thematic analysis of textual data revealed narratives which portray the perception of the society as a closed-door society, and described a clique culture in orthopaedics, and the pervasiveness of discrimination and banter. CONCLUSION: We found that barriers to inclusion and diversity exist within the professional society. Exploring the narratives around these has informed strategies to overcome them and has shaped future BHS initiatives. To ensure our patients receive the best possible surgical care, it is vital that those with the skills and expertise to deliver it, are supported by the Society and feel a sense of belonging and representation.

Histidine switch controlling pH-dependent protein folding and DNA binding in a transcription factor at the core of synthetic network devices.

Therapeutic strategies have been reported that depend on synthetic network devices in which a urate-sensing transcriptional regulator detects pathological levels of urate and triggers production or release of urate oxidase. The transcription factor involved, HucR, is a member of the multiple antibiotic resistance (MarR) protein family. We show that protonation of stacked histidine residues at the pivot point of long helices that form the scaffold of the dimer interface leads to reversible formation of a molten globule state and significantly attenuated DNA binding at physiological temperatures. We also show that binding of urate to symmetrical sites in each protein lobe is communicated via the dimer interface. This is the first demonstration of regulation of a MarR family transcription factor by pH-dependent interconversion between a molten globule and a compact folded state. Our data further suggest that HucR may be utilized in synthetic devices that depend on detection of pH changes.

Cost effectiveness of total hip arthroplasty in osteoarthritis: comparison of devices with differing bearing surfaces and modes of fixation.

Many different designs of total hip arthroplasty (THA) with varying performance and cost are available. The identification of those which are the most cost-effective could allow significant cost-savings. We used an established Markov model to examine the cost effectiveness of five frequently used categories of THA which differed according to bearing surface and mode of fixation, using data from the National Joint Registry for England and Wales. Kaplan-Meier analyses of rates of revision for men and women were modelled with parametric distributions. Costs of devices were provided by the NHS Supply Chain and associated costs were taken from existing studies. Lifetime costs, lifetime quality-adjusted-life-years (QALYs) and the probability of a device being cost effective at a willingness to pay £20 000/QALY were included in the models. The differences in QALYs between different categories of implant were extremely small (< 0.0039 QALYs for men or women over the patient's lifetime) and differences in cost were also marginal (£2500 to £3000 in the same time period). As a result, the probability of any particular device being the most cost effective was very sensitive to small, plausible changes in quality of life estimates and cost. Our results suggest that available evidence does not support recommending a particular device on cost effectiveness grounds alone. We would recommend that the choice of prosthesis should be determined by the rate of revision, local costs and the preferences of the surgeon and patient.

Effects of residual feed intake classification and method of alfalfa processing on ewe intake and growth.

The objective of this research was to evaluate the effects of residual feed intake (RFI) determined under ad libitum feeding conditions on DMI and performance of yearling ewes fed either chopped or pelleted alfalfa hay. In Exp. 1, 45 ewe-lambs had ad libitum access to a pelleted grower diet for 63 d and individual DMI was determined using an electronic feed delivery system. Residual feed intake values were assigned to each ewe-lamb as a measure of feed efficiency. Sixteen ewe-lambs with the most positive RFI values were classified as high RFI (inefficient) and 16 ewe-lambs with the most negative RFI values were classified as low RFI (efficient). In Exp. 2, half of the ewes from each efficiency group were placed into 1 of 2 pens and provided ad libitum access to either pelleted or chopped alfalfa hay. Individual DMI was again determined using an electronic feed delivery system. Body weight, LM area (LMA), and 12th-rib back fat thickness (BF) were measured at the beginning and end of both experiments. In Exp. 1, DMI by ewe-lambs in the low RFI group was 9% less (P = 0.01) than by ewe-lambs in the high RFI group (2.21 vs. 2.43 kg/d); however, ADG and initial and final BW, LMA, and BF did not differ (P > 0.27) among RFI groups. In Exp. 2, there were no feed processing × RFI group interactions (P > 0.14) for any trait. By design, RFI values were lower (P < 0.01) by yearling ewes in the low than high RFI group (-0.27 vs. 0.27); however, RFI values did not differ (P = 1.0) between yearling ewes fed chopped versus pelleted alfalfa. Dry matter intake was 22% less (P < 0.01) by yearling ewes in the low than high RFI group (2.5 vs. 3.2 kg/d) and 59% less (P < 0.01) by yearling ewes fed chopped versus pelleted alfalfa (2.2 vs. 3.5 kg/d). Initial and final BW, ADG, and G:F did not differ (P > 0.45) between RFI groups but were greater (P < 0.01) by yearling ewes fed pelleted alfalfa compared to chopped alfalfa. Final LMA did not differ (P = 0.77) between RFI groups, but final BF tended to be greater (P = 0.06) for high than low RFI yearling ewes (0.63 vs. 0.57 cm). Final LMA and BF were greater (P < 0.01) by yearling ewes fed pelleted than chopped alfalfa. Low RFI ewes had lower DMI and BF compared to high RFI ewes. Ewe efficiency as determined by RFI was repeatable on subsequent forage based diets; however, differences in intake and efficiency were more apparent when alfalfa was pelleted.

Adult granulosa cell tumor of the ovary: a retrospective study of 37 cases.

AIM: to evaluate the recurrence rate and the overall survival in women with adult granulosa cell tumor (AGCT), who were treated at the Department of Obstetrics and Gynecology, Aalborg University Hospital during the period January 1985 to January 2010. MATERIALS AND METHODS: Data from 38 women with AGCT were collected retrospectively. The histological slides were re-evaluated by a gynecologic pathologist. Surgical and pathological characteristics were analyzed. Results: Thirty-seven women with AGCT were diagnosed. 92% were diagnosed in FIGO Stage I and 8% in Stage II. The majority of patients (27 patients, 73%) were treated with total abdomi- nal hysterectomy and bilateral salpingooophorectomy. Only one patient received postoperative pelvic irradiation. The recurrence rate was 5.6%. CONCLUSION: The recurrent rate was 5.6%, which is low according to the literature. Primary surgical treatment with radical removal of tumor seemed to be appropriate treatment.

Clinical effectiveness and cost-effectiveness of second- and third-generation left ventricular assist devices as either bridge to transplant or alternative to transplant for adults eligible for heart transplantation: systematic review and cost-effectiveness model.

BACKGROUND: Advanced heart failure (HF) is a debilitating condition for which heart transplant (HT) offers the best treatment option. However, the supply of donor hearts is diminishing and demand greatly exceeds supply. Ventricular assist devices (VADs) are surgically implanted pumps used as an alternative to transplant (ATT) or as a bridge to transplant (BTT) while a patient awaits a donor heart. Surgery and VADs are costly. For the NHS to allocate and deliver such services in a cost-effective way the relative costs and benefits of these alternative treatments need to be estimated. OBJECTIVES: To investigate for patients aged ≥ 16 years with advanced HF eligible for HT: (1) the clinical effectiveness and cost-effectiveness of second- and third-generation VADs used as BTT compared with medical management (MM); and (2) the clinical effectiveness and cost-effectiveness of second- and third-generation VADs used as an ATT in comparison with their use as BTT therapy. DATA SOURCES: Searches for clinical effectiveness studies covered years from 2003 to March 2012 and included the following data bases: MEDLINE, MEDLINE In-Process & Other Non-Indexed Citations, EMBASE, Cochrane Database of Systematic Reviews (CDSR), Database of Abstracts of Reviews of Effects (DARE), NHS Economic Evaluation Database (NHS EED), HTA databases [NHS Centre for Reviews and Dissemination (CRD)], Science Citation Index and Conference Proceedings (Web of Science), UK Clinical Research Network (UKCRN) Portfolio Database, Cumulative Index to Nursing and Allied Health Literature (CINAHL), PsycINFO and National Library of Medicine (NLM) Gateway, Cochrane Central Register of Controlled Trials (CENTRAL), Current Controlled Trials and ClinicalTrials.gov. Reference lists of relevant articles were checked, and VAD manufacturers' websites interrogated. For economic analyses we made use of individual patient data (IPD) held in the UK Blood and Transplant Database (BTDB). REVIEW METHODS: Systematic reviews of evidence on clinical effectiveness and cost-effectiveness of second- and third-generation US Food and Drug Administration (FDA) and/or Conformité Européenne (CE) approved VADs. Publications from the last 5 years with control groups, or case series with 50 or more patients were included. Outcomes included survival, functional capacity (e.g. change in New York Heart Association functional classification), quality of life (QoL) and adverse events. Data from the BTDB were obtained. A discrete-time, semi-Markov, multistate model was built. Deterministic and probabilistic methods with multiple sensitivity analyses varying survival, utilities and cost inputs to the model were used. Model outputs were incremental cost-effectiveness ratios (ICERs), cost/quality-adjusted life-years (QALYs) gained and cost/life-year gained (LYG). The discount rate was 3.5% and the time horizon varied over 3 years, 10 years and lifetime. RESULTS: Forty publications reported clinical effectiveness of VADs and one study reported cost-effectiveness. We found no high-quality comparative empirical studies of VADs as BTT compared with MM or as ATT compared with BTT. Approximately 15-25% of the patients receiving a device had died by 12 months. Studies reported the following wide ranges for adverse events: 4-27% bleeding requiring transfusion; 1.5-40% stroke; 3.3-48% infection; 1-14% device failure; 3-30% HF; 11-32% reoperation; and 3-53% renal failure. QoL and functional status were reported as improved in studies of two devices [HeartMate II (HMII; Thoratec Inc., Pleasanton, CA, USA) and HeartWare (HW; HeartWare Inc., Framingham, MA, USA)]. At 3 years, 10 years and lifetime, the ICERs for VADs as BTT compared with MM were £122,730, £68,088 and £55,173 respectively. These values were stable to changes in survival of the MM group. Both QoL and costs were reduced by VADs as ATT compared with VADs as BTT giving ICERs in south-west quadrant of the cost effectiveness plain (cost saving/QALY sacrificed) of £353,467, £31,685 and £20,637 over the 3 years, 10 years and lifetime horizons respectively. Probabilistic analyses yielded similar results for both research questions. LIMITATIONS: Conclusions about the clinical effectiveness were limited by the lack of randomised controlled trials (RCTs) comparing the effectiveness of different VADs for BTT or comparing BTT with any alternative treatment and by the overlapping populations in published studies. Although IPD from the BTDB was used to estimate the cost-effectiveness of VADs compared with MM for BTT, the lack of randomisation of populations limited the interpretation of this analysis. CONCLUSIONS: At 3 years, 10 years and lifetime the ICERs for VADs as BTT compared with MM are higher than generally applied willingness-to-pay thresholds in the UK, but at a lifetime time horizon they approximate threshold values used in end of life assessments. VADs as ATT have a reduced cost but cause reduced QALYs relative to BTT. Future research should direct attention towards two areas. First, how any future evaluations of second- or third-generation VADs might be conducted. For ethical reasons a RCT offering equal probability of HT for each group would not be feasible; future studies should fully assess costs, long-term patient survival, QoL, functional ability and adverse events, so that these may be incorporated into economic evaluation agreement on outcomes measures across future studies. Second, continuation of accurate data collection in the UK database to encompass QoL data and comparative assessment of performance with other international centres. FUNDING: The National Institute for Health Research Health Technology Assessment programme.

Aspirin for prophylactic use in the primary prevention of cardiovascular disease and cancer: a systematic review and overview of reviews.

BACKGROUND: Prophylactic aspirin has been considered to be beneficial in reducing the risks of heart disease and cancer. However, potential benefits must be balanced against the possible harm from side effects, such as bleeding and gastrointestinal (GI) symptoms. It is particularly important to know the risk of side effects when aspirin is used as primary prevention--that is when used by people as yet free of, but at risk of developing, cardiovascular disease (CVD) or cancer. In this report we aim to identify and re-analyse randomised controlled trials (RCTs), systematic reviews and meta-analyses to summarise the current scientific evidence with a focus on possible harms of prophylactic aspirin in primary prevention of CVD and cancer. OBJECTIVES: To identify RCTs, systematic reviews and meta-analyses of RCTs of the prophylactic use of aspirin in primary prevention of CVD or cancer. To undertake a quality assessment of identified systematic reviews and meta-analyses using meta-analysis to investigate study-level effects on estimates of benefits and risks of adverse events; cumulative meta-analysis; exploratory multivariable meta-regression; and to quantify relative and absolute risks and benefits. METHODS: We identified RCTs, meta-analyses and systematic reviews, and searched electronic bibliographic databases (from 2008 September 2012) including MEDLINE, Cochrane Central Register of Controlled Trials, Database of Abstracts of Reviews of Effects, NHS Centre for Reviews and Dissemination, and Science Citation Index. We limited searches to publications since 2008, based on timing of the most recent comprehensive systematic reviews. RESULTS: In total, 2572 potentially relevant papers were identified and 27 met the inclusion criteria. Benefits of aspirin ranged from 6% reduction in relative risk (RR) for all-cause mortality [RR 0.94, 95% confidence interval (CI) 0.88 to 1.00] and 10% reduction in major cardiovascular events (MCEs) (RR 0.90, 95% CI 0.85 to 0.96) to a reduction in total coronary heart disease (CHD) of 15% (RR 0.85, 95% CI 0.69 to 1.06). Reported pooled odds ratios (ORs) for total cancer mortality ranged between 0.76 (95% CI 0.66 to 0.88) and 0.93 (95% CI 0.84 to 1.03). Inclusion of the Women's Health Study changed the estimated OR to 0.82 (95% CI 0.69 to 0.97). Aspirin reduced reported colorectal cancer (CRC) incidence (OR 0.66, 95% CI 0.90 to 1.02). However, including studies in which aspirin was given every other day raised the OR to 0.91 (95% CI 0.74 to 1.11). Reported cancer benefits appeared approximately 5 years from start of treatment. Calculation of absolute effects per 100,000 patient-years of follow-up showed reductions ranging from 33 to 46 deaths (all-cause mortality), 60-84 MCEs and 47-64 incidents of CHD and a possible avoidance of 34 deaths from CRC. Reported increased RRs of adverse events from aspirin use were 37% for GI bleeding (RR 1.37, 95% CI 1.15 to 1.62), between 54% (RR 1.54, 95% CI 1.30 to 1.82) and 62% (RR 1.62, 95% CI 1.31 to 2.00) for major bleeds, and between 32% (RR 1.32, 95% CI 1.00 to 1.74) and 38% (RR 1.38, 95% CI 1.01 to 1.82) for haemorrhagic stroke. Pooled estimates of increased RR for bleeding remained stable across trials conducted over several decades. Estimates of absolute rates of harm from aspirin use, per 100,000 patient-years of follow-up, were 99-178 for non-trivial bleeds, 46-49 for major bleeds, 68-117 for GI bleeds and 8-10 for haemorrhagic stroke. Meta-analyses aimed at judging risk of bleed according to sex and in individuals with diabetes were insufficiently powered for firm conclusions to be drawn. LIMITATIONS: Searches were date limited to 2008 because of the intense interest that this subject has generated and the cataloguing of all primary research in so many previous systematic reviews. A further limitation was our potential over-reliance on study-level systematic reviews in which the person-years of follow-up were not accurately ascertainable. However, estimates of number of events averted or incurred through aspirin use calculated from data in study-level meta-analyses did not differ substantially from estimates based on individual patient data-level meta-analyses, for which person-years of follow-up were more accurate (although based on less-than-complete assemblies of currently available primary studies). CONCLUSIONS: We have found that there is a fine balance between benefits and risks from regular aspirin use in primary prevention of CVD. Effects on cancer prevention have a long lead time and are at present reliant on post hoc analyses. All absolute effects are relatively small compared with the burden of these diseases. Several potentially relevant ongoing trials will be completed between 2013 and 2019, which may clarify the extent of benefit of aspirin in reducing cancer incidence and mortality. Future research considerations include expanding the use of IPD meta-analysis of RCTs by pooling data from available studies and investigating the impact of different dose regimens on cardiovascular and cancer outcomes. FUNDING: The National Institute for Health Research Health Technology Assessment programme.

Lean implementation in primary care health visiting services in National Health Service UK.

BACKGROUND: This paper presents the findings of a 13-month lean implementation in National Health Service (NHS) primary care health visiting services from May 2008 to June 2009. METHOD: Lean was chosen for this study because of its reported success in other healthcare organisations. Value-stream mapping was utilised to map out essential tasks for the participating health visiting service. Stakeholder mapping was conducted to determine the links between all relevant stakeholders. Waste processes were then identified through discussions with these stakeholders, and a redesigned future state process map was produced. Quantitative data were provided through a 10-day time-and-motion study of a selected number of staff within the service. This was analysed to provide an indication of waste activity that could be removed from the system following planned improvements. RESULTS: The value-stream map demonstrated that there were 67 processes in the original health visiting service studied. Analysis revealed that 65% of these processes were waste and could be removed in the redesigned process map. The baseline time-and-motion data demonstrate that clinical staff performed on average 15% waste activities, and the administrative support staff performed 46% waste activities. CONCLUSION: Opportunities for significant waste reduction have been identified during the study using the lean tools of value-stream mapping and a time-and-motion study. These opportunities include simplification of standard tasks, reduction in paperwork and standardisation of processes. Successful implementation of these improvements will free up resources within the organisation which can be redirected towards providing better direct care to patients.

Effect of intake level and alfalfa substitution for grass hay on ruminal kinetics of fiber digestion and particle passage in beef cattle.

Two experiments were conducted to evaluate digestion kinetics of alfalfa (Medicago sativa L.) substitution for grass hay in beef cattle. In Exp. 1, forage combinations evaluated in situ consisted of 0% alfalfa-100% big bluestem (Andropogon gerardi Vitman), 25% alfalfa-75% big bluestem, 50% alfalfa-50% big bluestem, and 100% alfalfa-0% big bluestem. Nonlinear regression was used to determine the immediately soluble fraction A, the potentially degradable fraction B, the undegraded fraction C, and the disappearance rate of DM and NDF. Dry matter fraction A increased linearly (P = 0.03), and DM and NDF fraction B decreased linearly (P = 0.01) with increasing alfalfa substitution. Rate of DM and NDF disappearance increased linearly (P /= 0.23) on total tract apparent digestibility of all nutrients except CP. Steers fed orchardgrass plus alfalfa had 33% greater (P = 0.01) total tract apparent digestibility for CP than those fed orchardgrass alone. Lag time of DM and NDF disappearance was not affected (P >/= 0.20) by alfalfa supplementation or intake level. Rate of DM and NDF disappearance of orchardgrass was faster (P

Telomerase activity determined by conventional telomeric repeat amplification protocol and reverse transcriptase-polymerase chain reaction assay in ovarian lesions: a comparison of assays.

OBJECTIVE: Telomerase is capable of restoring telomeric sequence lost during replication. No or low levels of telomerase activity are present in normal somatic cells, whereas up to 85-90% of all cancer cells express telomerase activity, suggesting telomerase as a possible tumor marker. The catalytic subunit, hTERT, correlates with the activity of the enzyme. MATERIAL AND METHODS: Telomerase activity in ovarian tissue was measured by the functional telomeric repeat amplification protocol (TRAP)eze assay, and a quantitative reverse-transcriptase-polymerase chain reaction (RT-PCR) assay measuring the expression of the telomerase catalytic subunit, hTERT. RESULTS: A weakly positive correlation was found between telomerase activity and severity of ovarian disease using the results of the TRAP assay, compared to a strongly positive correlation considering the results obtained in the RT-PCR assay. A statistically significant difference between the benign and borderline groups was present using the RT-PCR assay, allowing for screening for both borderline and primary malignant conditions with a specificity of 97% and a sensitivity of 68%. No significant statistical difference was found between telomerase activity in benign and borderline conditions when using the TRAP assay. When screening for primary malignancy, the specificity and sensitivity rates were 94% and 21%, respectively. CONCLUSIONS: The RT-PCR assay allowed discrimination between benign and borderline and malignant cases, and thereby proved superior to the TRAP assay, which could not discriminate the benign cases from the borderline cases. This suggests that the RT-PCR assay may be useful in screening for both borderline and primary malignancy in ovarian lesions.

Effect of cutting height and genetics on composition, intake, and digestibility of corn silage by beef heifers.

This research was conducted to determine the effect of corn genetics and cutting height on the composition and nutritive characteristics of corn silage. An in situ study involving eight commercially available corn hybrids indicated main effects and interactions (P < 0.01) of hybrid and cutting height on NDF, ADF, and starch content and on in situ DM and NDF degradablility. Four ruminally cannulated Angus heifers (initial BW = 378 +/- 3 kg) were used in a 4 x 4 Latin square digestion experiment with a 2 x 2 factorial treatment arrangement. Main effects and interactions of hybrid (Pioneer Hi-Bred Int., Inc., hybrids 3335 and 3223) and cutting height (LO = 20.3 cm, and HI = 61 cm) were evaluated. Dietary treatment consisted of 40% chopped alfalfa hay and 60% corn silage. Although corn silage hybrids used were of equivalent maturity at harvest (60% milkline), 3335 treatments had 37.8% starch and 34.8% NDF, whereas 3223 treatments had 33.7% starch and 38.6% NDF. The LO treatments averaged 3.1 percentage units greater in NDF and 3.45 percentage units less in starch content than the HI treatments. Intake of DM was greater for heifers fed 3335-HI than 3335-LO; however, DMI was greater by heifers fed 3223-LO than 3223-HI (hybrid x cutting height interaction, P < 0.05). Starch intake was greater (P < 0.05) and NDF intake was less (P < 0.05) by heifers fed HI vs. LO and fed 3335 vs. 3223 dietary treatments. Digestibility of DM, starch, and NDF was greater (P < 0.05) by heifers fed 3223 than 3335 dietary treatments, but digestibility differences were not observed (P > 0.10) between cutting heights. Rate of in situ DM and starch degradability was not affected (P > 0.10) by hybrid or cutting height; however DM degradability was greater (P < 0.05) for HI than LO corn silage substrates at 8, 16, and 24 h of incubation. Rate of NDF degradability tended (P = 0.08) to be greater for 3223 than for 3335, and for LO compared with HI corn silage. Degradability of NDF was greater (P < 0.05) for 3223 compared with 3335 substrates at 24, 36, and 48 h of incubation. These data suggest fiber may not be an accurate measure of corn silage quality. Whereas cutting height affected chemical composition, we observed genetics to have a greater effect on corn silage quality.

Chromosome 3 linked frontotemporal dementia (FTD-3).

BACKGROUND: The authors have identified and studied a large kindred in which frontotemporal dementia (FTD) is inherited as an autosomal dominant trait. The trait has been mapped to the pericentromeric region of chromosome 3. METHODS: The authors report on the clinical, neuroimaging, neuropsychological, and pathologic features in this unique pedigree collected during 17 years of study. RESULTS: Twenty-two individuals in three generations have been affected; the age at onset varies between 46 and 65 years. The disease presents with a predominantly frontal lobe syndrome but there is also evidence for temporal and dominant parietal lobe dysfunction. Late in the illness individuals develop a florid motor syndrome with pyramidal and extrapyramidal features. Structural imaging reveals generalized cerebral atrophy; H2 15 O-PET scanning in two individuals relatively early and late in the disease shows a striking global reduction in cerebral blood flow affecting all lobes. On macroscopic pathologic examination, there is generalized cerebral atrophy affecting the frontal lobes preferentially. Microscopically, there is neuronal loss and gliosis without specific histopathologic features. CONCLUSIONS: FTD-3 shares clinical and pathologic features with other forms of FTD and fulfills international consensus criteria for FTD. There is involvement of the parietal lobes clinically, radiologically, and pathologically in FTD-3 in contrast to some forms of FTD. This more diffuse involvement of the cerebral cortex leads to a distinctive, global pattern of reduced blood flow on PET scanning.

High-affinity DNA binding of HU protein from the hyperthermophile Thermotoga maritima.

Prokaryotic genomes are compacted by association with small basic proteins, generating what has been termed bacterial chromatin. The ubiquitous DNA-binding protein HU serves this function. DNA-binding properties of HU from the hyperthermophilic eubacterium Thermotoga maritima are shown here to differ significantly from those characteristic of previously described HU homologs. Electrophoretic mobility shift analyses show that T. maritima HU (TmHU) binds double-stranded DNA with high affinity (K(d)=5.6(+/-0.7) nM for 37 bp DNA). Equivalent affinity is observed between 4 degrees C and 45 degrees C. TmHU has higher affinity for DNA containing a set of 4 nt loops separated by 9 bp (K(d)=1.4(+/-0.3) nM), consistent with its introduction of two DNA kinks. Using DNA probes of varying length, the optimal binding site for TmHU is estimated at 37 bp, in sharp contrast to the 9-10 bp binding site reported for other HU homologs. Alignment of >60 HU sequences demonstrates significant sequence conservation: A DNA-intercalating proline residue is almost universally conserved, and it is preceded by arginine and asparagine in most sequences, generating a highly conserved RNP motif; V substitutes for R only in HU from Thermotoga, Thermus and Deinococcus. A fivefold increase in DNA-binding affinity is observed for TmHU in which V is replaced with R (TmHU-V61R; K(d)=1.1(+/-0.2) nM), but a change in the trajectory of DNA flanking the sites of DNA intercalation is inferred from analysis of TmHU-V61R binding to DNA modified with 4 nt loops or with substitutions of 5-hydroxymethyluracil for thymine. Survival in extreme environments places unique demands on protection of genomic DNA from thermal destabilization and on access of DNA to the cellular machinery, demands that may be fulfilled by the specific DNA-binding properties of HU and by the fine structure of the bacterial chromatin.

The ascertainment and management of tuberculosis in Tayside, Scotland during 1993-94.

OBJECTIVE: To evaluate whether utilising a database of dispensed prescriptions for anti-tuberculous chemotherapy could improve case ascertainment compared to standard methods. A further objective was to assess whether cases were managed according to standard guidelines. DESIGN: Retrospective case note audit. SETTING: Tayside, Scotland SUBJECTS: Patients identified by conventional (i.e. SMRI diagnostic codes, notifications to the Health Board, microbiology and pathology reports) and the data base (MEMO) as potentially having tuberculosis and receiving treatment between 1st January 1993 and 31st December 1994. One hundred and ninety one potential cases were identified. One hundred and twenty two case notes were obtained for review. Eighty eight of these were initially thought to have tuberculosis and the results below refer to these 88 cases. RESULTS: MEMO identified 43 cases not found by conventional methods. Cases identified by MEMO were more likely to have been managed as outpatients and less likely to have positive microbiology than cases identified by conventional means. Only 26 cases were notified to the Health Board, including all smear positive cases. CONCLUSIONS: Notification of tuberculosis continues to be incomplete. Use of the MEMO system almost doubled case ascertainment. The absence of a firm diagnosis may lead to a reluctance to notify cases being treated as tuberculosis.

Lymphangioleiomyomatosis. A series of three case reports illustrating the link with high oestrogen states.

Lymphangioleiornyomatosis is a rare lung disorder characterised by cystic air spaces and smooth muscle proliferation. The condition, which most commonly presents with dyspnoea, pneumothoraces or cough, is only described in females and is most commonly diagnosed during childbearing years. Three cases are presented which illustrate typical features of the disease and the association with high oestrogen levels. The first had recurrent pneumothoraces during her first pregnancy. The second lady was post menopausal at diagnosis but her symptoms predated her menopause. The third, presented with dyspnoea, abnormal chest sensations and a pneumothorax. She had a history of hyperprolactinaemia with secondary amenorhoea due to low oestrogen levels which had been corrected prior to her presentation. All three patients had reduced gas transfer and abnormalities in spirometry, two had reticular shadowing on their chest radiograph and all had typical appearances on lung computerised tomography. Although disease progression was variable, all patients showed a gradual decline in lung function.

Endothelial monocyte activating polypeptide II induces endothelial cell apoptosis and may inhibit tumor angiogenesis.

Endothelial monocyte activating polypeptide II (EMAP-II) is a tumor-derived cytokine with potent effects on endothelial cells in vitro and in vivo including upregulation of tissue factor and the sensitization of human melanoma to systemic TNF treatment via its effects on the tumor vasculature. We investigated the effects of EMAP-II on tumor growth, angiogenesis, vasculogenesis, and apoptosis. EMAP-II inhibited endothelial cell proliferation, vasculogenesis, and neovessel formation. In vivo growth of human melanoma lines expressing high amounts of EMAP-II demonstrated slower growth, smaller tumors, and increased amounts of tumor necrosis than those expressing lower amounts of EMAP-II. EMAP-II induced endothelial-cell-specific apoptosis via a pathway that includes upregulation of the Fas-associated death domain and downregulation of Bcl-2. EMAP-II appears to have important effects on angiogenesis and may play a role in regulating tumor vascular growth.

Split-night versus full-night studies for sleep apnoea/hypopnoea syndrome.

Investigation and treatment of sleep apnoea/hypopnoea syndrome (SAHS) is placing increasing demands on healthcare resources. This workload may be reduced by using split-night studies instead of the standard full-nights of diagnostic polysomnography and continuous positive airway pressure (CPAP) titration. Split-night studies involve polysomnography in the first half of the night followed, if there is an abnormal frequency of apnoeas and hypopneas, by CPAP titration for the remainder of the night. The authors' database of all patients prescribed a CPAP trial 1991-1997 was used to compare long-term outcomes in all 49 (46 accepting CPAP) patients prescribed split-night studies with those in full-night patients, matched 1:2 using an apnoea/ hypopnoea index (AHI) of +/-15% and Epworth score of +/-3 units. Classical symptoms of SAHS were the main reason for the split-night studies (n=27). There were no differences between the groups in long-term CPAP use, median nightly CPAP use (split-night 6.0 h x night-1, interquartile range (IQR) 3.8-7.4, full-night; 6.2 h x night-1, IQR 3.7-7.0, p=0.9), post-treatment Epworth scores and frequency of nursing interventions/clinic visits required. The median time from referral to treatment was less for the split-night patients (13 months, IQR 11-20 months) than for full-night patients (22 months, IQR 12-34 months; p=0.003). Split-night studies, in selected patients, result in equivalent long-term continuous positive airway pressure use to full-night studies with shorter treatment times and less healthcare utilization.

Mechanisms for the enhanced thermal stability of a mutant of transcription factor 1 as explained by (1)H, (15)N and (13)C NMR chemical shifts and secondary structure analysis.

A variant of the bacteriophage SPO1-encoded transcription factor 1 (TF1) with two site-specific mutations (E15G and T32I) was shown to be more thermally stable and bind DNA more tightly compared to the wild-type protein. In order to understand the biochemical mechanisms underlying these properties, we are engaged in determining the solution structures of this mutant alone and in complex with DNA using nuclear magnetic resonance (NMR) spectroscopy. The first phase of this project is reported here, as we have completed most of the backbone and sidechain sequential NMR assignments of the mutant protein, TF1-G15/I32. Insights derived from the (1)H, (15)N and (13)C chemical shifts and from the secondary structure analysis provide us with an explanation for the noted increase in thermal stability of TF1-G15/I32. Compared to the structure of the wild-type protein, the beta-sheet and the C-terminal helix remain largely unaffected whereas the mutations cause great changes in the first two helices and their enclosed loop. Specifically, we have found that the second helix is extended by one residue at its N-terminus and rotated in a way that allows Ala-37 to interact with Tyr-94 of the C-terminal helix. The loop has been found to become more rigid as a result of hydrophobic interactions between the flanking second and first helices and also between the second helix and the loop itself. Furthermore, the T32I mutation allows tighter packing between the second helix and the beta-sheet. Collectively, these changes contribute to a more tightly associated dimer and hence, to a greater thermal stability.

Differential protection by rat UDP-glucuronosyltransferase 1A7 against Benzo[a]pyrene-3,6-quinone- versus Benzo[a]pyrene-induced cytotoxic effects in human lymphoblastoid cells.

UDP-glucuronosyltransferase 1A7 (UGT1A7) is a polyaromatic hydrocarbon (PAH)-inducible UGT with activity toward various benzo[a]pyrene (B[a]P) metabolites. To investigate the influence of rat UGT1A7 on B[a]P-induced cytotoxicity, human lymphoblastoid L3 cells were transfected with pMF6 (control expression vector), p167Dtk2 (microsomal epoxide hydrolase expression vector), or p167Dtk2-1A7 (epoxide hydrolase/UGT1A7 coexpression vector), and the cell populations were compared for sensitivity to B[a]P-induced effects. B[a]P inhibited cell proliferation and decreased relative cell survival of p167Dtk2 and p167Dtk2-1A7 cells to a similar extent. Metabolism studies using [(3)H]B[a]P revealed increased formation of glucuronide conjugates of B[a]P-4,5-diol, 3-OH-, or 9-OH-B[a]P and an unidentified metabolite by p167Dtk2-1A7 cells, but the presence of unconjugated metabolites suggested that glucuronidation capacity may be limited. No differences between p167Dtk2 and p167Dtk2-1A7 L3 cells were observed in the growth inhibitory effects of 3-OH-B[a]P or B[a]P-7,8-diol, but p167Dtk2-1A7-expressing cells were found to be less sensitive to B[a]P-3,6-quinone-induced effects on cell proliferation and relative cell survival. The effect was also observed in AHH-1 lymphoblastoid cells expressing UGT1A7 without epoxide hydrolase. The UGT1A7-expressing AHH-1 cells were also less sensitive to growth inhibition by B[a]P-1,6-quinone and B[a]P-6,12-quinone. Flow cytometric analysis of vehicle and B[a]P-3, 6-quinone-exposed cell populations showed an association between UGT1A7 expression and resistance to B[a]P-3,6-quinone-induced apoptosis and loss of cell viability. These data suggest that UGT1A7 may be preferentially active toward B[a]P-quinones and that UGT1A7 may represent the PAH-inducible UGT activity previously implicated in protection against toxic redox cycling by B[a]P-3,6-quinone.