Hospital nurses' work environment, quality of care provided and career plans.

BACKGROUND: In several European countries, the availability of qualified nurses is insufficient to meet current healthcare requirements. Nurses are highly dissatisfied with the rising demands of the healthcare environment and increasingly considering leaving their jobs. AIM: The study aims to investigate the relationships between the characteristics of hospital nurses' work environment and the quality of care provided, and furthermore to examine Dutch nurses' career plans. METHODS: A cross-sectional, questionnaire survey of registered nurses (n = 334) working in the academic and district hospitals was conducted in 2005/2006. Previously validated questionnaires translated into the participants' language were used. Factor and regression analysis were used for data analysis. RESULTS: Overall, nurses rated their work environment rather favourably. Five work environment characteristics were identified: support for professional development, adequate staffing, nursing competence, supportive management and teamwork. Significant relationships were found between nurses' perceptions of their work environment characteristics and quality of care provided and nurses' career plans. When work environment characteristics were evaluated to be better, nurse-assessed quality of care also increased and intentions to leave current job decreased linearly. CONCLUSIONS: Study findings suggest that nurses' perceptions of their work environment are important for nurse outcomes in hospital settings. Further research is needed to explore the predictive ability of the work environment for nurse, patient and organizational outcomes in hospitals.

Determination of fungal succession during municipal solid waste composting using a cloning-based analysis.

AIMS: The microbiota at industrial full-scale composting plants has earlier been fragmentarily studied with molecular methods. Here, fungal communities from different stages of a full-scale and a pilot-scale composting reactors were studied before and after wood ash amendment. METHODS AND RESULT: The portion of fungal biomass, determined using phospholipid fatty acid analysis, varied between 6.3% and 38.5% in different composting phases. The fungal internal transcribed spacer (ITS) area was cloned and sequenced from 19 samples representing different stages of the composting processes. Altogether 2986 sequenced clones were grouped into 166 phylotypes from which 35% had a close match in the sequence databases. The fungal communities of the samples were related with the measured environmental variables in order to identify phylotypes typical of certain composting conditions. The fungal phylotypes could be grouped into those that dominated the mesophilic low pH initial phases (sequences similar to genera Candida, Pichia and Dipodascaceae) and those found mostly or exclusively in the thermophilic phase (sequences clustering to Thermomyces, Candida and Rhizomucor), but a few were also present throughout the whole process. CONCLUSIONS: The community composition was found to vary between suboptimally and optimally operating processes. In addition, there were differences in fungal communities between processes of industrial and pilot scale. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this study reveal the fungal diversity with molecular methods in industrial composting process. This is also one of the first studies conducted with samples from an industrial biowaste composting process.

Long-term sequelae of Farmer's lung disease in HRCT: a 14-year follow-up study of 88 patients and 83 matched control farmers.

The aim of this study was to compare high-resolution computed tomography (HRCT) findings of long-term farmer's lung (FL) patients and control farmers. We studied 88 FL patients and 83 matched control farmers with a mean follow-up time of 14 years. Emphysematous, fibrotic, and miliary changes were recorded by HRCT. The pattern of emphysema and location and distribution of other findings were evaluated in detail. Emphysema was found in 20 (23%) FL patients and in 6 (7%) controls (p=0.005). Recurrences of FL attacks increased (p=0.021) the risk of emphysema. Prevalence of fibrosis (17 vs 10%, p=0.16) and miliary changes (11 vs 4%, p=0.06) did not differ significantly in patients and controls. Among FL patients, emphysematous, fibrous, and miliary changes were more pronounced at the base than in the upper parts of the lung (p<0.02). In slice analysis, the pattern of emphysema was more polymorphous (p=0.001) and the distribution of fibrotic and miliary changes was more variable among FL patients than controls. Emphysema in HRCT is more common in FL patients than matched control farmers, and the occurrence is increased by recurrences of FL. Emphysematous, fibrous, and miliary changes in FL patients HRCT are multiform and predominate in the lower parts of the lung.

Evolution of MR contrast enhancement patterns during the first week after acute ischemic stroke.

BACKGROUND AND PURPOSE: Intravascular and parenchymal enhancement have been detected with contrast-enhanced T1-weighted MR imaging in patients with ischemic stroke. Diffusion-weighted MR imaging depicts infarct within minutes after the onset of symptoms. The aims of this study were to study the different MR enhancement findings during the first week after stroke and to ascertain whether the presence of intravascular enhancement over a larger area than the infarct on diffusion-weighted images on day 1 is able to predict substantial infarct growth during the first week. METHODS: Forty-eight patients were imaged on the first and second days and again 1 week after the onset of ischemic stroke. T1-weighted spin-echo imaging was performed before and after a 0.2 mmol/kg bolus of gadolinium chelate. Diffusion-weighted imaging was performed at the same slice positions. Enhancement findings were categorized as intravascular and parenchymal, with further categorization of parenchymal enhancement as cortical, subcortical, and deep; these findings were then compared with diffusion-weighted imaging findings. RESULTS: Intravascular enhancement in the infarcted area was detected on day 1 in 78% of the cases, on day 2 in 78% of the cases, and at 1 week in 30% of the cases. Parenchymal enhancement was detected in 26%, 56%, and 100% of the cases, respectively. Intravascular enhancement over a larger area than the infarct on diffusion-weighted images on day 1 was not associated with the extent of infarct growth. CONCLUSION: Detection of different patterns of contrast enhancement can help in determining the age of infarct. Parenchymal enhancement may be intense and can cause diagnostic uncertainty in cases in which the clinical history is obscure.

Combined perfusion- and diffusion-weighted MR imaging in acute ischemic stroke during the 1st week: a longitudinal study.

PURPOSE: To compare findings with different magnetic resonance (MR) perfusion maps in acute ischemic stroke. MATERIALS AND METHODS: Combined diffusion-weighted (DW) and perfusion-weighted (PW) MR imaging was performed in 49 patients with acute (<24 hours) stroke, on the 1st and 2nd days and 1 week after stroke. Volumes of hypoperfused tissue on maps of relative cerebral blood volume (rCBV), relative cerebral blood flow (rCBF), and mean transit time (MTT) were compared with the volume of infarcted tissue at DW imaging. RESULTS: The mean infarct volume increased from 41 to 65 cm(3) between the 1st and 2nd days (P: <.001; n = 49). On the 1st day, all perfusion maps on average showed hypoperfusion lesions larger than the infarct at DW imaging (P: <.001; n = 49). MTT maps showed significantly (P: <.001) larger hypoperfusion lesions than did rCBF maps, which showed significantly (P: <.001) larger hypoperfusion lesions than did rCBV maps. The sizes of the initial perfusion-diffusion mismatches correlated significantly with the extent of infarct growth (0.479 < r < 0.657; P:

Combined SPECT and diffusion-weighted MRI as a predictor of infarct growth in acute ischemic stroke.

UNLABELLED: In acute ischemic stroke, the infarcted core is surrounded by a zone of tissue that has decreased perfusion. Some of this tissue may be salvaged by prompt, effective treatment. Diffusion-weighted MRI is sensitive in detecting the infarcted tissue, whereas SPECT also detects the hypoperfused tissue around the infarcted core. We studied the potential of combined diffusion-weighted MRI and SPECT to predict infarct growth and clinical outcome in patients not receiving thrombolytic treatment. METHODS: Sixteen patients with acute stroke were examined consecutively with diffusion-weighted MRI and 99mTc-ethyl cysteinate dimer (99mTc-ECD) SPECT within 24 h of the onset of symptoms. Follow-up diffusion-weighted MRI was performed on the second day and after 1 wk. The volumes of infarcted and hypoperfused brain tissue were measured from diffusion-weighted MRI and SPECT, respectively. The volume difference between the hypoperfused and infarcted tissue on the first day was compared with the possible increase in infarct volume during the follow-up. Each patient's neurologic status was assessed with the National Institutes of Health Stroke Scale (NIHSS). RESULTS: The volume of infarcted tissue increased from 48 +/- 54 cm3 (mean +/- SD) on the first day to 88 +/- 93 cm3 on the second day (P = 0.001) and to 110 +/- 121 cm3 at 1 wk (P = 0.001). The volume of hypoperfused tissue on the first day was significantly greater than the infarct volume (102 +/- 135 cm3; P = 0.001). The volume difference between the hypoperfused and infarcted tissue on the first day correlated significantly with the infarct growth between the first day and 1 wk (r = 0.71; P < 0.01). Between the first day and 1 wk, the increase of the infarct volume correlated significantly with the change in the NIHSS (r = 0.54; P < 0.05). CONCLUSION: A large hypoperfusion zone around the infarct core in the acute phase of ischemic stroke predicts the infarct growth during the first week, and this correlates significantly with the change in the neurologic status of the patient. Combined diffusion-weighted MRI and SPECT performed within 24 h after the onset of symptoms can be useful in the evaluation of acute stroke to predict infarct growth.

Combined diffusion and perfusion MRI with correlation to single-photon emission CT in acute ischemic stroke. Ischemic penumbra predicts infarct growth.

BACKGROUND AND PURPOSE: More effective imaging methods are needed to overcome the limitations of CT in the investigation of treatments for acute ischemic stroke. Diffusion-weighted MRI (DWI) is sensitive in detecting infarcted brain tissue, whereas perfusion-weighted MRI (PWI) can detect brain perfusion in the same imaging session. Combining these methods may help in identifying the ischemic penumbra, which is an important concept in the hemodynamics of acute stroke. The purpose of this study was to determine whether combined DWI and PWI in acute (<24 hours) ischemic stroke can predict infarct growth and final size. METHODS: Forty-six patients with acute ischemic stroke underwent DWI and PWI on days 1, 2, and 8. No patient received thrombolysis. Twenty-three patients underwent single-photon emission CT in the acute phase. Lesion volumes were measured from DWI, SPECT, and maps of relative cerebral blood flow calculated from PWI. RESULTS: The mean volume of infarcted tissue detected by DWI increased from 46.1 to 75.6 cm(3) between days 1 and 2 (P<0.001; n=46) and to 78.5 cm(3) after 1 week (P<0.001; n=42). The perfusion-diffusion mismatch correlated with infarct growth (r=0. 699, P<0.001). The volume of hypoperfusion on the initial PWI correlated with final infarct size (r=0.827, P<0.001). The hypoperfusion volumes detected by PWI and SPECT correlated significantly (r=0.824, P<0.001). CONCLUSIONS: Combined DWI and PWI can predict infarct enlargement in acute stroke. PWI can detect hypoperfused brain tissue in good agreement with SPECT in acute stroke.

Cellular fibronectin concentration in the plasma of patients with malignant and benign diseases: a comparison with CA 19-9 and CEA.

EDAcFN enzyme immunoassay (EIA) is a new tumour marker assay measuring the extra domain A-containing isoform of cellular fibronectin (cFN), a component mainly found in extracellular matrices. The concentration cFN was measured in plasma and serum from 468 patients with malignant and benign diseases. The concentrations of cFN were higher in plasma than in serum. Using receiver operating characteristic (ROC) curve analysis, determination from plasma was superior to serum at specificity levels higher than 78% and was chosen for further analysis. The highest frequencies of elevated cFN values were seen in patients with hepato-pancreato-biliary malignancies (50-67%). In pancreatic and bile duct cancers, cFN provided little further information to that obtained by CA 19-9. The greatest advantage over CA 19-9 and CEA was seen in patients with local colorectal cancer and in hepatocellular carcinomas. Four out of nine patients with Dukes' stage B colorectal cancer had an elevated cFn level, but only one had an abnormal CEA level. In hepatocellular carcinomas, cFN was also compared with alpha-fetoprotein. The sensitivity of cFN (72%) was superior to that of AFP (61%), and concomitant use of cFN and AFP raised the sensitivity to 83%. The highest frequencies of elevated values in patients with benign diseases were observed in those with severe liver disease (32%) and biliary (17%) and pancreatic (24%) diseases. A combination of cFN and CA 19-9 showed the highest overall sensitivity of 47%, compared with 31% for cFN and 33% for CA 19-9. The corresponding specificities were 76% for cFN +/- CA 19-9, 85% for cFN and 83% for CA 19-9. The accuracy of a combination of cFN and CA 19-9 or CEA (60% respectively) was higher than that of cFN (55%), CA 19-9 (55%) or CEA (45%) alone. In conclusion, the results of the new cFN test are encouraging and further studies on larger patient materials have been started.

How should we estimate carotid stenosis using magnetic resonance angiography?

Our purpose was to assess the reproducibility of and differences between the most commonly used methods for assessing carotid artery stenosis using magnetic resonance angiography (MRA). We studied 55 patients who underwent axial three-dimensional time-of-flight MRA (1.5 T). Quantitative caliper measurements were performed from maximum intensity projection (MIP) and multiple planar reconstruction (MPR) images, according to the criteria of the North American Symptomatic Carotid Endarterectomy Trial (NASCET) and European Carotid Surgery Trial (ECST). The measurements were compared to each other and to visual interpretation, using conventional angiography as the reference. The measured percentage stenoses were higher on MRA than on digital subtraction angiography (DSA) using both NASCET (mean difference 1.9-3.0%) and ECST (6.3-6.7%) criteria. The kappa coefficients for the agreement between DSA and MRA were higher using the NASCET (0.61-0.76) than the ECST criteria (0.52-0.65). No statistically significant differences were found between measurements from MIP and MPR images. The ECST measurement criteria gave significantly higher percentage stenoses than the NASCET criteria (P < 0.001), this difference being more prominent on MRA (mean difference in diameter stenosis percentage 14.3-16.4%) than on DSA (7.6-11.2%) and most important with mild stenoses. The difference between visual interpretation and quantitative measurements on MRA was significant (P = 0.01-0.001). There were no statistically significant interobserver differences in the MRA film readings, either in visually estimated degrees of stenosis or stenosis measurements. Thus, the different criteria of the two multicentre trials led to significantly different results, especially in the assessment of mild stenosis, and these differences are more important with MRA than with DSA. Differences between the imaging modalities or the reconstruction programs seem less important.

Increased release of tenascin in tear fluid after photorefractive keratectomy.

BACKGROUND: Extracellular matrix protein tenascin (TN) is expressed in the anterior stroma during corneal wound healing. In this study we analysed TN release in tear fluid after photorefractive keratectomy (PRK). METHODS: Tear fluid TN concentrations of ten PRK patients were measured with an immunoassay. Tear fluids were collected preoperatively and 1, 2 and 7 days after PRK. The tear fluid collection time and the volume of tears collected were registered. Because tear fluid flow was greatly increased postoperatively, tear fluid flow-corrected release (TN flux) was calculated. RESULTS: The tear fluid flow was 4.50 +/- 0.94 microliters/min (mean +/- SEM) preoperatively, 55.48 +/- 16.70 microliters/min (P < 0.01) on the 1st, 33.91 +/- 7.91 microliters/min (P < 0.01) on the 2nd, and 13.79 +/- 5.49 microliters/min (P > 0.05) on the 7th postoperative day. The preoperative TN concentration was 0.85 +/- 0.20 microgram/ml. On the 1st postoperative day it decreased to 0.37 +/- 0.17 microgram/ml (P > 0.05), most likely due to the dilution effect caused by hypersecretion after PRK. The TN concentration was 0.67 +/- 0.12 microgram/ml (P > 0.05) on the 2nd and 0.78 +/- 0.15 micrograms/ml (P > 0.05) on the 7th postoperative day. The preoperative TN flux was 5.23 +/- 1.88 ng/min. On the 1st and 2nd postoperative days the TN flux was 14.40 +/- 4.99 ng/min (P < 0.05) and 22.66 +/- 6.12 ng/min (P < 0.05), respectively. On the 7th postoperative day a tendency towards decreased flux (14.00 +/- 6.02 ng/min, P > 0.05) was observed. CONCLUSION: Although there is a minor decrease in TN concentration after PRK due to increased tear fluid flow, a significant increase in TN flux was observed. Complete reepithelialization of the ablated area was observed in all eyes at the follow-up visit on postoperative day 7.

Enzyme immunoassay for quantification of tenascin in biologic samples.

An enzyme immunoassay was developed for quantification of tenascin in biologic samples. An enzyme conjugate prepared by coupling peroxidase to a well-characterized, affinity-purified monoclonal antibody EB2 to human tenascin was used as principal reagent. The assay comprises 96-well microtitration strip plates with immobilized monoclonal antibody DB7 to human tenascin. By using a novel monoclonal antibody suppressing human-anti-mouse-factor, MAK33, in the sample buffer, the specificity of the test could be improved. The method has a minimum detectable sensitivity of 1.5 ng tenascin and permits determination of tenascin in various biologic samples. The coefficients of variation within run and between run ranged from 0.9% to 5.0%. The average tenascin concentration in normal plasma was 0.97 mg/L (n = 200) and in serum 0.73 mg/L (n = 200). The tenascin concentrations were also determined in samples of urine, bile, amniotic fluid, seminal fluid, cerebrospinal fluid, bronchoalveolar lavage (BAL) fluid, and pleural fluid showing general applicability of the assay. The method permits the determination of tenascin in samples of different body fluids from various diseases, including cancer, showing increased amounts of the protein at the tissue level.

Increased release of immunoreactive calcitonin gene-related peptide (CGRP) in tears after excimer laser keratectomy.

The purpose of the study was to quantify the neuropeptide calcitonin gene-related peptide (CGRP) in normal human tear fluid and to determine the effect of photorefractive excimer laser keratectomy (PRK) on its release in tears. CGRP was assayed in tear fluid samples using an enzyme immunoassay (detection limit 0.2 micrograms ml-1). Tear-fluid samples were collected preoperatively, 1, 2 and 7 days after PRK and analysed for CGRP. The changes in tear-fluid secretion were also monitored. The intra-assay variation was 3.0-7.0%. Despite the marked hypersecretion of tears, the concentration of CGRP did not decrease following PRK indicating a concomitant increase in CGRP release by sensory nerves and/or lacrimal gland(s). Consequently, the release of CGRP in tears increased from 197.9 +/- 36.6 ng min-1 (mean +/- S.E.M.) to 1723.0 +/- 402.4 ng min-1 (P < 0.01) on day 1, and to 2304.2 +/- 561.1 ng min-1 (P < 0.01) on day 2. On day 7, only minor elevation (377.02 +/- 83.24 ng min-1) was observed. It is concluded that CGRP is a component of normal human tear fluid. The ocular irritation response related to the photoablation induces an enhanced release of CGRP in tears. As a compound present in corneal sensory nerves CGRP may have a role in wound-healing.

Intracranial aneurysms: MR angiographic screening in 400 asymptomatic individuals with increased familial risk.

PURPOSE: To evaluate the accuracy and limitations of magnetic resonance (MR) angiographic screening for incidental intracranial aneurysms (IAs) in healthy individuals with a family history of IA. MATERIALS AND METHODS: Four hundred asymptomatic individuals in 68 families with a history of aneurysmal subarachnoid hemorrhage underwent MR angiography. Six families also had a history of polycystic kidney disease. A multislab, three-dimensional, time-of-flight sequence was used. Conventional angiography was performed to confirm positive MR angiographic findings. RESULTS: IAs were detected with MR angiography in 37 individuals, 32 of whom underwent conventional angiography. Intraobserver consistency was excellent (kappa = 0.96), and interobserver reproducibility was good to excellent (kappa = 0.59-0.82). Both techniques showed similar results in the evaluation of size, localization, and orientation of aneurysms and visibility of the aneurysm neck. CONCLUSION: MR angiography is an accurate and feasible method of noninvasive screening for IAs in individuals at high risk. Conventional angiography is still necessary before operative treatment.

Cellular fibronectin in serum and plasma: a potential new tumour marker?

The concentration of cellular fibronectin (cFN) containing the extra domain A (EDA) was measured in 479 plasma and 300 serum samples from healthy blood donors by a competitive enzyme immunoassay (EIA). Serum and plasma samples contained low concentrations of EDAcFN. The mean concentration of EDAcFN was higher in plasma (2.46 mg l-1) than in serum (0.30 mg l-1). No significant differences between sexes or age groups were found. The EDAcFN concentrations were also measured in 120 patients with various malignancies. The mean values in serum were 4.28 mg l-1, 2.01 mg l-1 and 5.18 mg l-1 in patients with digestive tract malignancies, breast cancer and a group of miscellaneous cancers respectively. In plasma, the corresponding values were 12.26 mg l-1, 4.38 mg l-1 and 11.12 mg l-1 respectively. The serum EDAcFN concentration was higher than the 97.5th percentile level of healthy blood donors in 86% of patients with digestive tract and in 76% with miscellaneous malignancies. In patients with breast cancer 60% had elevated levels of EDAcFN. The corresponding figures for plasma samples in patients with digestive tract and miscellaneous malignancies were 79% and 71% respectively. In patients with breast cancer only 30% had elevated plasma levels of EDAcFN. The mean values in serum and plasma of 20 patients with benign diseases were below the cut-off levels. Consistent with the EIA results, Western blotting revealed increased amounts of EDAcFN in blood samples from cancer patients. Pregnancy did not affect the EDAcFN concentration. The mean values in 20 pregnant women were below the cut-off levels.

Tear fluid cellular fibronectin levels after photorefractive keratectomy.

BACKGROUND: Fibronectin is supposed to have an important role in wound healing. The extradomain A-containing cellular fibronectin (EDAcFn) refers to fibronectin, which instead of being a hepatocyte derived component of blood plasma or body fluids, is produced locally. The present study was undertaken to clarify the possible changes in excretion of EDAcFn in tears following excimer laser photorefractive keratectomy (PRK). METHODS: An immunoassay was used to determine EDAcFn concentrations in human tear fluid samples of 11 eyes after PRK. Tear fluids were collected with scaled microcapillaries preoperatively as well as 1, 2, and 7 days after PRK. The time used to collect a known volume of tears was registered. This was done to estimate the dilution effect related to the hypersecretion of tears after PRK. RESULTS: The mean preoperative tear fluid EDAcFn concentration was 0.28 +/- 0.07 ng/microliter with a wide range (0.05 to 0.63). The tear fluid flow-corrected excretion of EDAcFn was 1.36 +/- 0.35 ng/min (range, 0.145 to 3.50). There was a significant increase in both postoperative tear fluid flow and excretion of EDAcFn on days 1 and 2. The elevation of the mean EDAcFn concentration did not decrease in spite of reflex tearing. The mean excretion of EDAcFn in tears was 28-fold on the first and 17-fold on the second postoperative day. Normal level was reached by day 7. CONCLUSION: There is a rapid increase in excretion of EDAcFn in tears following PRK. This seems to last only as long as an epithelial defect persists. The epithelium of all eyes healed in 3 to 4 days in spite of wide interindividual variations in both tear fluid flow and EDAcFn excretion.

Release of calcitonin gene-related peptide in tears after excimer laser photorefractive keratectomy.

BACKGROUND: Sensory nerves known to affect corneal healing are damaged to a variable degree after photorefractive keratectomy (PRK). To search for nerve-bound factors involved in corneal healing, we monitored tear fluid calcitonin gene-related peptide (CGRP) levels of six human eyes undergoing PRK. METHODS: CGRP concentrations were determined using an immunoassay. RESULTS: Normal human tear fluid contains CGRP. The mean CGRP concentration was slightly increased postoperatively, despite a marked tear fluid hypersecretion. Consequently, an almost ten-fold increase in release of CGRP in tears was observed on days 1 and 2 after PRK. Values measured on day 7 had returned to a normal level. CONCLUSION: The observed postoperative increase in release of CGRP in tears may have an impact on the healing of PRK wounds. Extensive neural damage following deep photoablations may impair healing and should probably be avoided.

Carotid artery stenosis: clinical efficacy of MR phase-contrast flow quantification as an adjunct to MR angiography.

PURPOSE: To assess the utility of magnetic resonance (MR) flow quantification in the evaluation of suspected carotid artery stenosis. MATERIALS AND METHODS: Fifty-five patients referred for angiography and 10 healthy volunteers underwent Doppler ultrasound, three-dimensional time-of-flight MR angiography, and MR phase-contrast flow quantification to measure peak systolic velocity (PSV) and volumetric flow rate (VFR) in the common and internal carotid arteries distal to the stenosis. RESULTS: PSV and VFR were significantly lower in the severely (> or = 70%) stenosed internal carotid arteries (P < .05). The VFR ratio (internal carotid artery-common carotid artery) achieved 91% overall accuracy for detection of severe stenosis. Combination of flow data with results of MR angiography yielded sensitivity of 100% (95% confidence interval, 78%-100%) with a modest loss in specificity. Doppler and MR measurements of PSV in the common carotid artery showed significant correlation: volunteers, r = .73; patients, r = .64. CONCLUSION: MR flow quantification provides information about the hemodynamic significance of carotid stenosis. As an adjunct to MR angiography, it may be useful in enabling differentiation of occlusions from critical stenoses.

An improved method for quantification of extra domain A-containing cellular fibronectin (EDAcFN) in different body fluids.

A quantitative direct enzyme immunoassay for the extra domain A-containing isoform of cellular fibronectin (EDAcFN) was established for screening of large series of blood samples and various body fluids of different pH and viscosity. The method is based on the monoclonal antibody DH1 recognizing the extra domain A in cellular fibronectin (EDAcFN). Studies on the effect of dilution of plasma and serum samples in this direct assay indicated that the measured concentration of cFN in the samples greatly depend on the ratio of sample dilution. The linearity of the assay was improved with sample dilution and the optimal dilution was 1:5. Stored diluted samples retained their cFN content at +4 degrees C, and -20 degrees C and -70 degrees C for months in contrast to samples stored undiluted. With this direct EIA the detection limit was 0.05 micrograms/ml and the linear portion of the standard curve could be extended above 30 micrograms/ml. Thus, the cFN concentration of blood samples could be measured reliably without inhibition also in samples with very high concentration of cFN. This is particularly important when measuring blood samples from cancer patients, since these samples may contain more than 20 micrograms/ml EDAcFN. The assay was standardized for blood samples but, due to the possibility of sample dilution, it also enabled reliable quantification of EDAcFN in various other body fluids. Undiluted some of the samples with non-neutral pH (urine, bile) or with high viscosity (seminal plasma) interfered with the assay. In addition to blood samples, the EDAcFN concentration was determined in samples of urine, bile, amniotic fluid, cervicovaginal secretions, seminal fluid, cerebrospinal fluid, bronchoalveolar lavage fluid, pleural fluid and saliva. Thereby, this modified method was shown to be applicable to various body fluids.

Comparison of ultrathin x-ray window designs.

In order to fabricate entrance windows for soft x-ray detectors, various technologies have been developed. Depending on the x-ray-detector type and the environment in which the windows are used, entrance windows must meet several, often contradictory, requirements: while good pressure tolerance and durability as well as gas tightness require thicker structures, good x-ray transmission can only be achieved with thin membranes. In this paper, the suitability of different window types for various applications is discussed. The applicability discussion is based on the results of tests performed on prototype windows, as well as on calculated and measured x-ray transmission properties. A comparative study of endurance vs transmission properties of some candidate membrane materials is also presented. Test results include pressure tolerance and leakage rates as well as some measurements of radiation damage to the window materials. The window technologies presented include coated polyimide membranes with two different supporting schemes as well as submicrometer beryllium membranes.

Competitive enzyme immunoassay for quantification of the cellular form of fibronectin (EDAcFN) in blood samples.

A competitive enzyme immunoassay based on the monoclonal antibody DH1 was developed for the quantification of extradomain A (EDA)-containing isoform of cellular fibronectin (cFN). The average EDAcFN concentration in normal plasma was 2.46 micrograms/ml and in serum 0.68 micrograms/ml. Similar results were obtained by gelatin-Sepharose binding and immunoblotting. Studies on the effect of storage of plasma and serum as frozen samples indicated that the amount of EDAcFN decreased rapidly when stored at -20 degrees C. Storage at -70 degrees C resulted in less of a decrease in the concentration of EDAcFN. The method permits the determination of EDAcFN in blood samples from diseases showing increased amounts of the protein at the tissue level.