The prevalence of an elevated F cell population in a maternal and gynaecology cohort.

OBJECTIVES: This study aimed to determine F cell prevalence in a cohort of maternal and gynaecology specimens using QuikQuant anti-HbF flow cytometry (FC) kit and to investigate if the presence of maternal F cells can lead to fetomaternal haemorrhage (FMH) overestimation. BACKGROUND: The gold standard to estimate FMH is the Kleihauer-Betke test (KBT). The KBT has proved to be insufficiently sensitive to detect low numbers of circulating fetal cells due to the presence of maternal F cells. At present, the prevalence of false positive KBT results due to raised maternal F cell population, defined as >5%, is poorly characterised. METHODS: A total of 120 specimens were tested for the presence of F cells and fetal cells by KBT and anti-HbF FC. The results calculated were compared to determine FMH overestimation. RESULTS: Of our cohort, 32% showed an elevated F cell population, of which 69% (27 of 39) were clinically significant according to KBT (>2 mL FMH). The mean FMH volumes by KBT and anti-HbF FC were 3·90 mL (0·20-35·40 mL) and 4·09 mL (0·20-9·70 mL), respectively. CONCLUSION: The study highlighted that an elevated F cell level could be found in the cohort tested, with an F cell level of >10% causing significant FMH overestimation by KBT.

Effectiveness of CHA2DS2-VASc based decision support on stroke prevention in atrial fibrillation: A cluster randomised trial in general practice.

BACKGROUND: Guidelines on atrial fibrillation (AF) recommend the CHA2DS2-VASc rule for anticoagulant decision-making, but underuse exists. We studied the impact of an automated decision support on stroke prevention in patients with AF in a cluster randomised trial in general practice. METHODS: Intervention practices were provided with a CHA2DS2-VASc based anticoagulant treatment recommendation. Reference practices provided care as usual. The primary outcome was incidence of ischaemic stroke, transient ischaemic attack (TIA) and/or thromboembolism (TE). Secondary outcomes were bleeding and the proportion of patients on guideline recommended anticoagulant treatment. RESULTS: In total, 1129 AF patients were included in the 19 intervention practices and 1226 AF patients in the 19 reference practices. The median age was 77 (interquartile range (IQR) 68-75) years, the median CHA2DS2-VASc score was 3.0 (IQR 2.0-5.0). Underuse of anticoagulants in patients with CHA2DS2-VASc score ≥ 2 was 6.6%. After a median follow-up of 2.7 years (IQR 2.3-3.0), the incidence rate per 100 person-years of ischaemic stroke/TIA/TE was 1.96 in the intervention group and 1.42 in the reference group (hazard ratio (HR) 1.3, 95% C.I. 0.8-2.1). No difference was observed in the rate of bleeding (0.79 versus 0.82), or in the underuse (7.2% versus 8.2%) or overuse (8.0% versus 7.9%) of anticoagulation. CONCLUSIONS: In this study in patients with AF in general practice, underuse of anticoagulants was relatively low. Providing practitioners with CHA2DS2-VASc based decision support did not result in a reduction in stroke incidence, affect bleeding risk or anticoagulant over- or underuse.

The efficacy of fibrinogen concentrate compared with cryoprecipitate in major obstetric haemorrhage--an observational study.

BACKGROUND: Fibrinogen replacement is critical in major obstetric haemorrhage (MOH). Purified, pasteurised fibrinogen concentrate appears to have benefit over cryoprecipitate in ease of administration and safety but is unlicensed in pregnancy. In July 2009, the Irish Blood Transfusion Service replaced cryoprecipitate with fibrinogen. OBJECTIVES: To examine the impact of this externally imposed change on blood product use and clinical outcomes in MOH. METHODS: Women with MOH requiring fibrinogen between 1 January 2009 and 30 June 2011 were identified from an MOH database. Aetiology of MOH, medical treatments, blood product use and clinical outcomes were compared between the cryoprecipitate and fibrinogen groups. RESULTS: Of 21 614 deliveries, 77 cases of MOH were identified. Of the 77 cases, 34 (44%) received cryoprecipitate (n = 14) or fibrinogen concentrate (n = 20). The mean (± SEM) dose utilised was 2.21 ± 0.35 pools of cryoprecipitate and 4 ± 0.8 g of fibrinogen. There was a stronger correlation between the increase in fibrinogen level and dose of fibrinogen (Pearson co-efficient 0.5; P = 0.03) than dose of cryoprecipitate (Pearson co-efficient 0.32; P = 0.3). Mean (± SEM) estimated blood loss (EBL), red cell concentrate (RCC) and Octaplas transfused were greater (but not significantly) in the cryoprecipitate group compared with the fibrinogen group; EBL = 5.2 ± 1.1 vs 3.3 ± 0.5 L (P = 0.1); RCC = 7.2 ± 1.2 vs 5.9 ± 1.0 U (P = 0.4); Octaplas = 4.1 ± 0.7 vs 3.2 ± 0.7 U (P = 0.36), respectively. Haemostasis was secured, and there were no adverse reactions or thrombotic complications. CONCLUSION: Purified virally inactivated fibrinogen concentrate is as efficacious as cryoprecipitate in correcting hypofibrinogenaemia in MOH.

Correlation of the BRAF V600E mutation in hairy cell leukaemia with morphology, cytochemistry and immunophenotype.

Hairy cell leukaemia (HCL) has distinct clinical, morphological and immunophenotypic features with no recurrent cytogenetic or molecular abnormalities reported until the recent description of the BRAF V600E mutation in patients with classical HCL. The incidence of this mutation was sought in 27 patients with either classical HCL or HCL variant by an allele-specific PCR approach and findings related to morphology, cytochemistry and immunophenotype. A high degree of correlation was noted between the presence of BRAF V600E and established diagnostic criteria in 26/27 patients with HCL/HCL variant. Detection of the BRAF V600E mutation is therefore a useful adjunct in the differential diagnosis of HCL and HCL variant and highlights the value of a multifaceted approach to the diagnosis of this malignancy.

Construction and elementary mode analysis of a metabolic model for Shewanella oneidensis MR-1.

A stoichiometric model describing the central metabolism of Shewanella oneidensis MR-1 wild-type and derivative strains was developed and used in elementary mode analysis (EMA). Shewanella oneidensis MR-1 can anaerobically respire a diverse pool of electron acceptors, and may be applied in several biotechnology settings, including bioremediation of toxic metals, electricity generation in microbial fuel cells, and whole-cell biocatalysis. The metabolic model presented here was adapted and verified by comparing the growth phenotypes of 13 single- and 1 double-knockout strains, while considering respiration via aerobic, anaerobic fumarate, and anaerobic metal reduction (Mtr) pathways, and utilizing acetate, n-acetylglucosamine (NAG), or lactate as carbon sources. The gene ppc, which encodes phosphoenolpyruvate carboxylase (Ppc), was determined to be necessary for aerobic growth on NAG and lactate, while not essential for growth on acetate. This suggests that Ppc is the only active anaplerotic enzyme when cultivated on lactate and NAG. The application of regulatory and substrate limitations to EMA has enabled creation of metabolic models that better reflect biological conditions, and significantly reduce the solution space for each condition, facilitating rapid strain optimization. This wild-type model can be easily adapted to include utilization of different carbon sources or secretion of different metabolic products, and allows the prediction of single- and multiple-knockout strains that are expected to operate under defined conditions with increased efficiency when compared to wild type cells.

Quality of care in the management of major obstetric haemorrhage.

Substandard care is reported to occur in a large number of cases of major obstetric haemorrhage (MOH). A prospective audit was carried out by a multidisciplinary team at our hospital over a one year period to assess the quality of care (QOC) delivered to women experiencing MOH. MOH was defined according to criteria outlined in the Scottish Audit of Maternal Morbidity (SAMM). 31 cases were identified yielding an incidence of 3.5/1000 deliveries. The predominant causes were uterine atony 11 (35.4%), retained products of conception 6 (19.3%) and placenta praevia/accreta 6 (19.3%). Excellent initial resuscitation and monitoring was noted with a high level of senior staff input. Indicators of QOC compared favourably with the SAMM. Areas for improvement were identified. This pilot study demonstrates the feasibility of detailed prospective data collection in MOH in a busy Dublin obstetric unit with a view to developing a national audit. Standardization of definitions allows for international comparisons.

Haemoglobin Etobicoke, an incidental finding in an Irish diabetic.

It is well recognized that haemoglobin variants can be detected during the measurement of HbA1c by high-performance liquid chromatography (HPLC). A number of variants have been reported as compromising the quantification of HbA1c, a marker used in the assessment of glycaemic control in diabetes. We describe a case of haemoglobin Etobicoke, a rare alpha chain variant detected in an Irish diabetic during HbA1c analysis. Its identity was confirmed using a series of investigations. These included haemoglobin electrophoresis at alkaline and acid pH, isoelectric focusing and globin chain electrophoresis. Ultimately mass spectrometry isolated the mutation at position alpha 84 (F5). Haemoglobin Etobicoke, first described in Canada in 1969 has not previously been detected on HbA1c analysis. In the presence of this rare variant, HbA1c, a standard method using HPLC to assess glycaemic control in diabetes is unreliable and alternatives such as fructosamine need to be considered. HbA1c measured by automated HPLC will effectively screen populations where haemoglobin variants were not previously known. Precise identity of these variants when they are detected is crucial to the reliable interpretation of HbA1c analyses.

Asymptomatic lung carcinoma presenting with a large vulvar lesion.

OBJECTIVE: Metastases to the vulva are infrequent, accounting for less than 10% of vulvar tumors. Vulvar metastases from lung carcinoma have been reported in two other cases. METHODS: A case of lung carcinoma metastatic to the vulva is reported. RESULTS: A 71-year-old woman was referred to The University of Iowa Hospitals and Clinics with a lung nodule on chest x-ray and a 12-cm, necrotic, left vulvar mass. A lung biopsy showed poorly differentiated carcinoma, and a palliative resection of the vulvar mass showed metastatic lung carcinoma. The patient died from sepsis on postoperative day 10. CONCLUSIONS: Lung carcinoma metastatic to the vulva is rare and portends a poor prognosis.

Biomarker distribution after injection into the canine prostate: implications for gene therapy.

OBJECTIVE: To evaluate the distribution of biomarkers after transrectal injection into the canine prostate and to report a method for enhancing the distribution of gene expression. MATERIALS AND METHODS: Carbon black was first used to evaluate the histopathological distribution in canine prostate of single or multiple injections via the transurethral, transperineal and transrectal routes. The distribution of canarypox virus (ALVAC) vector-delivered gene expression was then compared using both fluid-phase injection techniques and delivery in a solid carrier composed of a gelatine sponge matrix. RESULTS: After transurethral administration, carbon black was detected as scattered particles in ducts and acini, mostly in the periphery of the gland. Direct transrectal injection of carbon black resulted in a localized collection at the site of injection, with only a minimal peri-acinar distribution. Transrectal injection of the fluid-phase (virus suspended in diluent) ALVAC vector encoding the beta-galactosidase gene resulted in a similar distribution, with limited gene expression at the site of injection and in the needle track. Delivery of the same number of virus particles in the gelatine sponge matrix resulted in qualitatively greater gene expression. CONCLUSIONS: Direct injection of the canine prostate with biomarkers, including viral vectors, in the fluid-phase results in very localized gene expression, while the distribution was more widespread after delivery in a gelatine sponge matrix.

Effect of clinical history on diagnostic accuracy in the cytologic interpretation of bronchial brush specimens.

There has been little study of the effect of clinical history on pathologic diagnostic accuracy. Five pathologists retrospectively examined 97 bronchial brush specimens with and without clinical historic information. Forty-nine patients had a biopsy-proven malignant lesion, and 48 had a benign lesion. Diagnostic accuracy with and without history for each pathologist was determined with likelihood ratios and receiver operating characteristic curves. The overall diagnostic accuracy with and without history was 0.84 and 0.76, respectively. The average negative predictive value of a benign diagnosis decreased from 89.2% (with history) to 74.0% (without history). Overall, the cytopathologists were more reluctant to make a definitive malignant diagnosis without history compared with history. The average positive predictive value of a malignant diagnosis with and without history was almost identical. The absence of history leads to lower diagnostic accuracy in the cytologic interpretation of bronchial brush specimens partly because pathologists underdiagnose malignant lesions.

Application of the Etest to the antimicrobial susceptibility testing of Mycobacterium marinum clinical isolates.

Mycobacterium marinum, a well-recognized cutaneous pathogen, is usually treated by chemotherapy without available standardized in vitro susceptibility testing information. In this study, we have attempted to apply the stable-gradient method (Etest; AB Biodisk, Solna, Sweden) to susceptibility testing of M. marinum in order to assess the activities of eight antimicrobial agents against 60 recent clinical strains of M. marinum collected from 10 geographic sites within the United States. Two plated media (5% sheep blood Mueller-Hinton agar and Middlebrook 7H11 agar) were compared, and 7H11 agar was found to be superior in supporting the growth of all strains. Four reference strains of M. marinum were tested on five occasions with eight drugs (160 tests) in order to evaluate Etest reproducibility. Results were observed to be within 1 log2 dilution of the all-test median MIC for 97.5% of the Etests. Our MIC results for the 60 strains clearly demonstrate the best in vitro potency against M. marinum isolates to be as follows (rank order): trimethoprim-sulfamethoxazole (MIC at which 90% of the isolates are inhibited [MIC90], 0.25 and 4.25 microg/ml, respectively) = ethambutol > clarithromycin (MIC90, 1 microg/ml) > minocycline = doycycline (MIC90, 4 microg/ml) > amikacin (MIC90, 8 microg/ml). Rifampin was only marginally active against the M. marinum strains tested (MIC90, at the National Committee for Clinical Laboratory Standards) breakpoint of 1 microg/ml), and ciprofloxacin was not active (MIC90, 8 microg/ml). These data should enhance the empiric drug selection for contemporary M. marinum infections and also provide evidence that the Etest can be utilized to guide chemotherapy with alternative agents.

In vitro efficacy of levofloxacin alone or in combination tested against multi-resistant Pseudomonas aeruginosa strains.

Levofloxacin, the S(-) isomer of ofloxacin, demonstrates in vitro activity against Pseudomonas aeruginosa. To further characterize this activity, levofloxacin was tested against three populations of recent clinical isolates categorized by their resistance patterns to several other anti-pseudomonal agents. Results demonstrate the minimum inhibitory concentrations (MICs) for levofloxacin were generally two- to fourfold higher than for ciprofloxacin. Higher fluoroquinolone MICs were associated with MIC increases in other drugs. Levofloxacin demonstrated cross resistance against ciprofloxacin-resistant strains. Combinations of levofloxacin and several codrugs revealed that the majority of evaluable interactions demonstrated indifferent action. Levofloxacin exhibited enhanced activity (additive or degrees of synergy) principally with piperacillin, aztreonam, or ceftazidime. The synergy and additive rate (21 to 30%) compared favorably with the enhanced interactions observed with gentamicin combined with piperacillin or ceftazidime (27 to 30%). Levofloxacin activity against P. aeruginosa was most comparable to that of ciprofloxacin, which was applicable against > 90% of strains found to be resistant to other classes of antimicrobial agents.