Utilizing quantitative dried blood spot analysis to objectively assess adherence to cardiovascular pharmacotherapy among patients at Kenyatta National Hospital, Nairobi, Kenya.

The burden of cardiovascular disease (CVD) is rising in Kenya and non-adherence to cardiovascular pharmacotherapy is a growing global public health issue that leads to treatment failure, an increased risk of cardiac events and poor clinical outcomes. This study assessed adherence to selected cardiovascular therapy medications among CVD patients attending outpatient clinics at Kenyatta National Hospital, Kenya by determining drug concentration(s) in patient dried blood spot (DBS) samples. Patients who had been taking one or more of the five commonly prescribed CVD medications (amlodipine, atenolol, atorvastatin, losartan, and valsartan) for at least six months were enrolled. Each patient completed a short questionnaire about their medication history and then provided a finger-prick blood spot sample from which drug concentrations were determined by liquid chromatography-high resolution mass spectrometry analysis. Two hundred and thirty-nine patients (62.3% female) participated in the study. The median number of medications used by patients was 2 (IQR 75%-25% is 3-1). Less than half (117; 49.0%) of patients were adherent to their prescribed CVD pharmacotherapy. Binary regression analysis revealed a significant correlation between non-adherence and the number of medications in the treatment regimen (Odds Ratio (OR) 1.583; 95%CI: 0.949-2.639; P-value = 0.039) and that gender was not an independent predictor of medication adherence (OR 1.233; 95%CI: 0.730-2.083; P-value = 0.216). Valuable information about adherence to each medication in the patient's treatment regimen was obtained using quantitative DBS analysis showing that adherence to CVD medications was not uniform. DBS sampling, due its minimally invasive nature, convenience and ease of transport is a useful alternative matrix to monitor adherence to pharmacotherapies objectively, when combined with hyphenated mass spectrometry analytical techniques. This information can provide physicians with an evidence-based novel approach towards personalization and optimization of CVD pharmacotherapy and implementing interventions in the Kenyan population, thereby improving clinical outcomes.

Notchplasty is associated with decreased risk of anterior cruciate ligament graft revision.

PURPOSE: Despite influencing knee biomechanics and outcomes, the use of notchplasty at time of anterior cruciate ligament reconstruction (ACLR) has not been evaluated with regards to risk of secondary injury and revision. This study evaluates this association. METHODS: 42 patients (21.7-years, IQR = 19.0-27.5) that underwent primary then revision ACLR at a single institution were contrasted with a case matched control group of patients with grafts that did not fail. Patients were propensity score matched in a 1:2 ratio by age, gender, and date of index procedure. Post-hoc statistical correction was made for post-index procedure sports participation level. RESULTS: Notchplasty was performed in 2 of 42 cases that went on to revision, and in 31 of 84 cases in the control group (p < 0.001). This was associated with reduced rates of revision ACLR (OR = 0.085, 95%CI = 0.019-0.378). A significant difference was seen in the post-ACLR activity level between groups (p = 0.028), with post-hoc testing highlighting those returning to competitive sport to be more likely to require subsequent revision (OR = 9.647, 95%CI = 1.947-47.795). Notchplasty remained significantly associated with (reduced) risk of revision surgery, despite the observed variation in post-ACLR activity (p = 0.001). CONCLUSION: Individuals whose graft failed following ACLR were significantly less likely to have had notchplasty performed as part of their surgery than a control group who did not suffer graft reinjury. We propose that this may be due to decreased tensioning of the graft as the knee enters dynamic valgus, which may be of great relevance to athletes undergoing ACLR to enable return to sport.

Functional outcomes and return to sport following anterior cruciate ligament reconstruction in recreational athletes: A systematic review.

INTRODUCTION: Outcomes of anterior cruciate ligament reconstruction (ACLR) are well reported in athletic populations, however surprisingly little information is available for the recreational athletes that make up the majority of cases. The aim was therefore to assess post-operative outcome and return-to-sport in recreational athletes following ACLR. METHODS: A systematic search was conducted in Ovid MEDLINE, CINAHL, AMED and the grey literature according to PRISMA guidelines. Studies involving a clear definition of recreational athletes who underwent ACLR and recorded postoperative outcomes were included. Publication quality was assessed using Newcastle-Ottawa Scale. RESULTS: 107 studies were identified, 19 full-text records reviewed and 13 included, reflecting 1342 patients with an average age of 31.7 (SD 9.8) years. Mean follow-up was 43.6 (SD 42.8) months. Activity change post-surgery was reported in 92% (12/13) papers. Outcomes were assessed with the Tegner score in seven studies, four of which reported pre-injury scores, which worsened from 5.4 to 4.3 at final follow-up (76.5 months). 54% (7/13) studies reported return to pre-injury level of sport. In these, 59% (n = 327/555) achieved pre-injury level at a mean follow-up of 33.7 months (SD 38.6). The return-to-sports rate increased with length of follow-up. Methodological quality was moderate. CONCLUSION: Substantial variation in the timeframes and outcomes assessed restricts pooled analysis of change in function. Based on seven studies, 59% of recreational athletes return to pre-injury level of sport following ACLR. The link between return rate and post-operative review timeframe suggests that longer follow-up may be required to capture return-to-sport rates in this population.

Radiographic severity, extent and pattern of cartilage loss are not associated with patient reported outcomes before or after total knee arthroplasty in end-stage knee osteoarthritis.

BACKGROUND: The aim of this study was to determine if radiographic severity, extent or pattern of knee osteoarthritis was associated with pain and function before total knee arthroplasty (TKA) or improvement therein one year after TKA. METHODS: A prospective study of 259 patients undergoing unilateral TKA for Kellgren-Lawrence (KL) grade ≥ 3 knee osteoarthritis was conducted: mean age 69.8 ± 9.7 (44-91); mean BMI 31.0 ± 5.8 (17-52); 152/259 (58.7%) female. Preoperative radiographs were assessed using the KL and Ahlback systems. Preoperatively and 1 year postoperatively patients completed Oxford Knee Scores, VAS-Pain and EQ-5D scores. Full thickness cartilage loss was recorded intraoperatively. RESULTS: Median radiographic severity was Ahlback 2, KL 4: 51/259 (19.7%) hypertrophic; 23/259 (8.8%) atrophic. Neither Ahlback nor Kellgren-Lawrence OA grade was associated with OKS, VAS Pain or EQ-5D prior to TKA (p > 0.05). The extent and pattern of cartilage loss did not affect preoperative PROMs. Radiographic OA severity, compartment involvement, and pattern of cartilage loss were not significantly associated with PROMs or improvements therein following TKA (p > 0.05). Hypertrophic OA was associated with less pain before TKA (difference 6.8, 0.23-13.9 95%CI, p = 0.044), and worse improvement in OKS following TKA (difference -3.41, -6.8 to -0.05 95%CI, p = 0.047). Better preoperative OKS and hypertrophic OA were independently associated with poorer improvement in OKS 1 year following TKA (R2 = 0.208). CONCLUSION: Provided at least one compartment has KL grade ≥ 3 changes, further radiographic severity, pattern or extent of cartilage loss did not affect PROMs before or after TKA: multicompartmental was no worse than unicompartmental disease.

Adherence to cardiovascular pharmacotherapy by patients in Iraq: A mixed methods assessment using quantitative dried blood spot analysis and the 8-item Morisky Medication Adherence Scale.

This study evaluated the adherence to prescribed cardiovascular therapy medications among cardiovascular disease patients attending clinics in Misan, Amara, Iraq. Mixed methods were used to assess medication adherence comprising the Arabic version of the eight-item Morisky Medication Adherence Scale (MMAS-8) and determination of drug concentrations in patient dried blood spot (DBS) samples by liquid chromatography-high resolution mass spectrometry. Three hundred and three Iraqi patients (median age 53 years, 50.5% female) who had been taking one or more of the nine commonly prescribed cardiovascular medications (amlodipine, atenolol, atorvastatin, bisoprolol, diltiazem, lisinopril, losartan, simvastatin and valsartan) for at least six months were enrolled. For each patient MMAS-8 scores were determined alongside drug concentrations in their dried blood spot samples. Results from the standardized questionnaire showed that adherence was 81.8% in comparison with 50.8% obtained using the laboratory-based microsample analysis. The agreement between the indirect (MMAS-8) and direct (DBS analysis) assessment approaches to assessing medication adherence showed significantly poor agreement (kappa = 0.28, P = 0.001). The indirect and direct assessment approaches showed no significant correlation between nonadherence to prescribed cardiovascular pharmacotherapy and age and gender, but were significantly associated with the number of medications in the patient's treatment regimen (MMAS-8: Odds Ratio (OR) 1.947, 95% CI, P = 0.001; DBS analysis: OR 2.164, 95% CI, P = 0.001). The MMAS-8 results highlighted reasons for nonadherence to prescribed cardiovascular pharmacotherapy in this patient population whilst the objective DBS analysis approach gave valuable information about nonadherence to each medication in the patient's treatment regimen. DBS sampling, due its minimally invasive nature, convenience and ease of transport is a useful alternative matrix to monitor adherence objectively in Iraq to cardiovascular pharmacotherapy. This information combined with MMAS-8 can provide clinicians with an evidence-based novel approach to implement intervention strategies to optimise and personalise cardiovascular pharmacotherapy in the Iraqi population and thereby improve patient health outcomes.

Hyphenated mass spectrometry techniques for assessing medication adherence: advantages, challenges, clinical applications and future perspectives.

Nonadherence to prescribed pharmacotherapy is an understated public health problem globally and is costing many patients their chance to return to good health and healthcare systems billions. Clinicians need an accurate assessment of adherence to medications to aid the clinical decision-making process in the event of poor patient progress and to maximise the patient health outcomes from the drug therapies prescribed. An overview of indirect and direct methods used to measure medication adherence is presented, highlighting the potential for accurate measuring of drugs in biological samples using hyphenated mass spectrometry (MS) techniques to provide healthcare professionals with a reliable evidence base for clinical decision making. In this review we summarise published applications of hyphenated MS techniques for a diverse range of clinical areas demonstrating the rise in the use of such direct methods for assessing medication adherence. Although liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods using plasma, serum and urine samples are the most popular, in recent years increased attention has been given to liquid chromatography high-resolution mass spectrometry (LC-HRMS) methods and alternative biosample matrices including hair, saliva and blood microsamples. The advantages and challenges of using hyphenated MS techniques to address this healthcare problem are also discussed alongside future perspectives.

A review of Fourier Transform Infrared (FTIR) spectroscopy used in food adulteration and authenticity investigations.

The increasing demand for food and the globalisation of the supply chain have resulted in a rise in food fraud, and recent high profile cases, such as the Chinese milk scandal in 2008 and the EU horsemeat scandal in 2013 have emphasised the vulnerability of the food supply system to adulteration and authenticity frauds. Fourier Transform Infrared (FTIR) spectroscopy is routinely used in cases of suspected food fraud as it offers a rapid, easy and reliable detection method for these investigations. In this review, we first present a brief summary of the concepts of food adulteration and authenticity as well as a discussion of the current legislation regarding these crimes. Thereafter, we give an extensive overview of FTIR as an analytical technique and the different foods where FTIR analysis has been employed for food fraud investigations as well as the subsequent multivariate data analyses that have been applied successfully to investigate the case of adulteration or authenticity. Finally, we give a critical discussion of the applications and limitations of FTIR, either as a standalone technique or incorporated in a test battery, in the fight against food fraud.

The impact of a preoperative information leaflet on expectation management, satisfaction and patient outcomes in patients undergoing knee arthroscopy.

BACKGROUND: This study examined the effects of a patient information leaflet on outcomes related to patient satisfaction following knee arthroscopy. METHODS: Cohort study of patients listed for knee arthroscopy under the care of a single surgeon over a nine-month period (May 2017-January 2018) following the introduction of an information leaflet as an adjunct to the consent process. Outcome data was collected postoperatively through telephone follow-up. Outcome measures included feelings of involvement with decision-making, expectations being met, satisfaction, postoperative pain numerical rating scales and the Forgotten Joint Score-12. RESULTS: Fifty-five patients were consented by the operating surgeon, of which 28 (50.9%) received a leaflet and 27 (49.1%) did not. Patients who received the information leaflet felt more involved in and informed about the decision to have an operation than patients who did not (p = 0.016), however there were no differences in any other outcomes between patients who did and did not receive a leaflet (p > 0.05). CONCLUSIONS: The use of an information leaflet as an adjunct to the preoperative consultation is an effective way of helping patients feel more involved in the surgical decision-making process, however this does not influence overall outcome or satisfaction metrics.

Quantitative screening of the pharmaceutical ingredient for the rapid identification of substandard and falsified medicines using reflectance infrared spectroscopy.

The World Health Organization suggests that approximately 10% of medicines worldwide are either falsified or substandard with higher figures in low and middle income countries. Such poor quality medicines can seriously harm patients and pose a threat to the economy worldwide. This study investigates attenuated total reflectance-fourier transform infrared (ATR-FTIR) spectroscopy as a simple and rapid method for determination of drug content in tablet dosage forms. Paracetamol was used as the model pharmaceutical ingredient. Spectra of standard mixtures of paracetamol with different excipients formed the basis for multivariate PLS based quantitative analysis of simulated tablet content using different selected infrared absorbance bands. Calibration methods using ATR-FTIR were compared with the ATR-FTIR and conventional ultraviolet spectroscopic analyses of real tablet samples and showed that the paracetamol/microcrystalline cellulose mixtures gave optimum results for all spectral bands tested. The quantitative data for band 1524-1493cm-1 was linear (R2 ˃ 0.98; LOQ ≥ 10%w/w tablet). Global examples of paracetamol tablets were tested using this protocol and 12% of the tablet samples examined was identified as substandard. Each sample analysis was completed in just a few minutes. ATR-FTIR can therefore be used in the rapid screening of tablet formulations. The simplicity of the proposed method makes it appropriate for use in low and middle income countries where analytical facilities are not available.

Quantitative LC-HRMS determination of selected cardiovascular drugs, in dried blood spots, as an indicator of adherence to medication.

Dried blood spot (DBS) sampling was investigated as a means of obtaining micro-volume blood samples for the quantitative analyses of ten commonly UK prescribed cardiovascular drugs as an indicator of medication adherence. An 8mm disc was punched out from each DBS from calibration, quality control and volunteer samples and extracted using methanol containing the internal standard. Each extract was evaporated to dryness, the residue reconstituted in methanol:water (40:60v/v) containing 0.1% formic acid and analysed by LC-HRMS. Chromatography was performed using gradient elution on a Zorbax Eclipse C18 HD 100mm×2.1mm, 1.8µm pore size column with the column oven temperature at 40°C. Flow rate of the mobile phase was 0.6ml/min with a run time of 2.5min. Electrospray positive ionization was used for MS detection. Drug recoveries from spiked blood spots were 68% for simvastatin and ≥87% for all other target drugs. Compound specificity was obtained operating the MS with a 5ppm mass window. The LC-HRMS method was validated, with results for accuracy and precision within acceptable limits; analytes were stable at room temperature for at least 10 weeks and different blood spot volumes and haematocrit values had no significant effect. The LC-HRMS assay was used to analyse DBS samples from volunteers, some of whom were prescribed one or more of the target drugs. In results from 37 volunteers the assay successfully identified volunteers who were known to be either adherent or nonadherent; confirmed the correct drug/drugs for multiple prescriptions; demonstrated no false positives from other cardiovascular drugs; revealed several examples of unsuspected non-adherence. These results indicated that the developed assay was suitable for trials with patients.

Dried blood spot analysis to assess medication adherence and to inform personalization of treatment.

Little research using dried blood spot samples to assess adherence to medication has been reported. The World Health Organisation estimates that only half of the patients in the developed world take their medication as prescribed. Additional costs to the healthcare provider include wasted medicines, avoidable additional hospital visits and non-optimum patient care. There is little evidence of information concerning medication adherence being made available to inform clinical decision making. In this article we explore the potential of the dried blood spot sample collection methodology as a means of identifying medication adherence to facilitate medicines optimization for a range of disparate diseases. Furthermore, the opportunity to personalize healthcare for different patients by assessing the clinically necessary therapeutic level of the relevant drugs is highlighted.

Accelerated polyethylene wear and early failure of the uncemented Reflection acetabular component: a ten to fifteen-year follow-up study.

PURPOSE: The Reflection® second-generation uncemented acetabular component was designed to address increased rates of failure observed with the early acetabular designs. However, the reported survivorship of this acetabular component has been conflicting. The aim of this study was to describe the ten to 15-year survivorship and polyethylene wear rate for the uncemented Reflection® acetabular component performed as part of a primary total hip replacement. METHODS: One hundred and four consecutive Reflection® uncemented acetabular components in 97 patients were identified from a prospective arthroplasty database with a minimum of ten years of follow-up. No patient was lost to follow-up. Mean cohort age was 59.1 years. RESULTS: There were 24 revisions of the acetabular component. The all-cause survival rate at ten years was 77.2% [95% confidence interval (CI) 73.9-80.5]. Mean linear wear was 0.20 mm [standard deviation (SD) 0.08] per year and the volumetric wear was 106.2 mm(3) per year. At final follow-up, more than half the patients had osteolysis identified around the femoral component. The mean Oxford Hip Score was 35.6 (SD 9.8) at a mean follow-up of 11.9 years, with six (11.1%) excellent, 26 (48.1%) good, 17 (31.5%) fair and five (9.3%) poor outcomes. Despite the fact that more than a third had a fair or poor outcome, only four (7.4%) were not satisfied with their hip. CONCLUSIONS: Due to the high rate of relatively asymptomatic polyethylene wear and osteolysis associated with this acetabular component, in our department, we now review all surviving patients both clinically and radiographically on an annual basis.

Bisoprolol, ramipril and simvastatin determination in dried blood spot samples using LC-HRMS for assessing medication adherence.

The use of dried blood spot (DBS) collection cards was investigated for the quantification of three therapeutic drugs used in cardiovascular therapy for assessing medication adherence. A liquid chromatography-high resolution mass spectrometry (LC-HRMS) method was developed and validated for the determination of bisoprolol, ramipril and simvastatin. Whole blood spiked with target analytes was used to produce 30 µl blood spots on specimen collection cards. An 8mm disc was cut from the dried blood spot and extracted using methanol: water (70:30, v/v) containing the internal standard, atenolol. Extracts were vortexed, sonicated and then centrifuged. Gradient chromatographic elution was achieved using a Zorbax Eclipse C18 HD 100 mm × 2.1 mm, 1.8 µm pore size column and a mobile phase flow rate of 0.6 ml/min and the column oven temperature at 40 °C with a run time of 3 min. MS detection was carried out in electrospray positive ion mode for the three target drugs and for the IS. Drug recoveries from spiked blood spots were ≥ 92% for bisoprolol and ramipril and ~43% for simvastatin and the drugs were stable in DBS for at least 12 weeks. Validation of the LC-HRMS method showed good linearity and the accuracy (relative error) and precision (coefficient of variation) values were within the pre-defined limits of ≤ 15% at all concentrations. Matrix effects and the effects of different volumes of blood applied to the collection card were investigated. The LC-HRMS method successfully identified control volunteers who were known to be either adherent or non-adherent. There were no false positives from volunteers taking other cardiovascular drugs or from volunteers receiving no medication.

Quantitative determination of atenolol in dried blood spot samples by LC-HRMS: a potential method for assessing medication adherence.

The use of blood spot collection cards was investigated as a means of obtaining small volume samples for the quantification of therapeutic drugs for assessing medication adherence. A liquid chromatography-high resolution TOF mass spectrometry (LC-HRMS) method, based on the measurement at the accurate mass to charge ratio of the target analyte, was used to ensure specificity for atenolol in the dried blood spot (DBS) samples. A working method was developed and validated. For the preparation of DBS samples whole blood spiked with analyte was used to produce 30 µl blood spots on specimen collection cards. A 5mm disc was cut from the dried blood spot and extracted using methanol:water (60:40, v/v) containing the internal standard, atenolol-d(7). Extracts were vortexed, sonicated and then centrifuged. Gradient chromatographic elution was achieved using an Ascentis Express C18 100mm×2.1mm column and a mobile phase flow rate of 0.2 ml/min and the column oven temperature at 30 °C. MS detection was carried out in electrospray positive ion mode for target ions at accurate mass m/z 267.1703 for atenolol and 274.2143 for the IS. Drug extraction efficiency from spiked blood spots was demonstrated to be 96±5% and the drug was stable in DBS for at least 10 weeks. The developed LC-HRMS method was linear within the tested calibration range of 25-1500 ng/ml and validation showed the accuracy (relative error) and precision (coefficient of variation) values were within the pre-defined limits of ≤ 5% at all concentrations with a limit of quantification of 25 ng/ml. Factors with potential to affect drug quantification measurements such as the matrix effects, volume of blood applied onto the collection card and effect of different sampling cards were investigated. The developed LC-HRMS method was applied to blood spots on sampling card taken from adult healthy volunteers previously administered a 50mg atenolol tablet and a DBS concentration-time profile was obtained for atenolol. Requiring only a micro volume (30 µl) blood sample for analysis, the developed DBS based assay has the potential to assess patient adherence to atenolol.

Dexamethasone quantification in dried blood spot samples using LC-MS: The potential for application to neonatal pharmacokinetic studies.

A high-performance liquid chromatography (LC-MS) method has been developed and validated for the determination of dexamethasone in dried blood spot (DBS) samples. For the preparation of DBS samples whole blood spiked with analyte was used to produce 30µl blood spots on specimen collection cards. An 8mm disc was cut from the DBS sample and extracted using a combination of methanol: water (70:30, v/v) containing the internal standard, triamcinolone acetonide. Extracts were centrifuged and chromatographic separation was achieved using a Zorbax Eclipse Plus C18 column using gradient elution with a mobile phase of acetonitrile and water with formic acid at a flow rate of 0.2ml/min. LC-MS detection was conducted with single ion monitoring using target ions at m/z 393.1 for dexamethasone and 435.1 for the internal standard. The developed method was linear within the tested calibration range of 15-800ng/ml. The overall extraction recovery of dexamethasone from DBS samples was 99.3% (94.3-105.7%). The accuracy (relative error) and precision (coefficient of variation) values were within the pre-defined limits of ≤15% at all concentrations. Factors with potential to affect drug quantification measurements such as blood haematocrit, the volume of blood applied onto the collection card and spotting device were investigated. Although a haematocrit related effect was apparent, the assay accuracy and precision values remained within the 15% variability limit with fluctuations in haematocrit of ±5%. Variations in the volume of blood spotted did not appear to affect the performance of the developed assay. Similar observations were made regarding the spotting device used. The methodology has been applied to determine levels of dexamethasone in DBS samples collected from premature neonates. The measured concentrations were successfully evaluated using a simple 1-compartment pharmacokinetic model. Requiring only a microvolume (30µl) blood sample for analysis, the developed assay is particularly suited to pharmacokinetic studies involving paediatric populations.

The theory-practice gap: ECMO research example.

AIM: The time taken for research findings to enter clinical practice can be very lengthy. A contributing factor is the time lag between the research literature identifying the issue, and medical and nursing texts discussing it. The example of ECMO (Extracorporeal Membrane Oxygenation) is used to examine this issue, specifically the extent to which relevant nursing journals and textbooks discuss the effects of ECMO on medication. METHOD: A systematic review of papers identifying the problem (pharmacokinetics in ECMO patients) and the dissemination of this to clinicians was undertaken. Publications used by those exploring the problem were most likely to be found in Medline, and those disseminating to nurses in CINAHL. Textbooks on neonatal nursing and paediatric intensive care with sections on ECMO were also explored. RESULTS: There are several studies that show drug delivery is altered in patients receiving ECMO, dating back to 1989. Only three papers likely to be accessed by clinicians were found to address the effect of ECMO on drug delivery. Two of the textbooks addressed these issues but it took eight years from the issue being first raised to any medical or nursing text discussing it. CONCLUSION: Nurses will most likely not be aware of the latest research in their area if they rely on textbooks. Advanced education that gives critical appraisal and literature-searching skills, such as found in master's courses, should help the clinical nurse employ evidence based practice.

Analysis of post-Byzantine icons from the Church of the Assumption in Cephalonia, Ionian Islands, Greece: a multi-method approach.

A multi-method approach has been developed for the characterisation of the proteinaceous binding media, drying oils and pigments present in samples from the panel paintings of the Church of the Assumption in Cephalonia (Ionian Islands, Greece). The analytical protocol involved the use of scanning electron microscopy/energy dispersive X-ray analysis (SEM/EDX), Raman spectroscopy and gas chromatography. The identification of the pigments was achieved by SEM/EDX and Raman spectroscopy. The latter technique was also used for the detection of the binding media, while their characterisation was achieved by gas chromatographic analysis of ethyl chloroformate derivatives. The aim of this multi-method protocol was to obtain as much information as possible from the panel paintings of the Church of the Assumption, through non-destructive methods, before proceeding to gas chromatography. Little scientific information is available for the understanding of the construction technique and the materials used by the post-Byzantine artists and whatever is available comes mainly from artists' manuals. One of the aims of this paper is to provide a scientific background to the technology of the Ionian post-Byzantine icons.

Prevalence of increased serum creatine kinase activity due to macro-creatine kinase and experience of screening programmes in district general hospitals.

BACKGROUND: Macro creatine kinase type 1 (MCK1) may be the cause of elevated total serum CK activity, which can lead to diagnostic confusion. There is evidence that this problem is poorly recognized perhaps due to a lack of information on its prevalence. Precipitation with polyethylene glycol (PEG) has been described for the detection of MCK1 but has not been fully evaluated. METHODS: We introduced a screening programme to detect elevated serum total CK due to MCK1 and determine the prevalence of this problem using PEG precipitation with confirmation by gel filtration chromatography (GFC). The results were compared with those from a laboratory which selected samples for further investigation during the clinical validation process. We also studied characteristics of the PEG precipitation test including sensitivity and specificity when compared with GFC. RESULTS: Over 2 years we screened 368 patients. In 17 cases the proportion of CK activity precipitated by PEG was relatively high and the presence of MCK1 was confirmed in seven by GFC. In a second laboratory, over a period of 5 years, 11 samples were selected during the clinical validation process for further study and MCK1 was the cause of the elevated CK activity in six cases. PEG precipitates a proportion of normal, uncomplexed CK and this is increased by increasing serum globulin concentration and by higher concentrations of PEG. CONCLUSIONS: The prevalence of elevated serum CK activity due to MCK1 was approximately 2%. Laboratories should consider introducing a systematic screening programme based on PEG precipitation.

Evidence of cardiomyocyte necrosis in glycogen storage disease type II.

Adult-onset glycogen storage disease type II (GSD-II), unlike the infantile form, is not normally associated with coexisting cardiovascular pathologies. In infantile onset GSD-II, cardiomyopathy is a common feature, and mutations in the genes for cardiac troponin T and I are likely to be involved. This case report describes a 39-year-old man with no classical risk factors for premature cardiac disease who presented with central chest pain and shortness of breath. Serum aspartate transaminase (AST) had been consistently elevated for 15 years. Adult GSD-II had been diagnosed two years previously by muscle biopsy. On presentation, there was an elevated serum creatine kinase and AST. Electrocardiography and echocardiography were both normal, and an acute coronary syndrome was ruled out. Cardiac Troponin T (cTnT) was found to be positive at 0.1 microg/L using a Cardiac Reader, subsequently confirmed on an Elecsys 1010 (both from Roche Diagnostics, Lewes, UK). cTnT may therefore be a useful biomarker in examining subclinical cardiac involvement in GSD-II patients.