COMP: a candidate molecule in the pathogenesis of systemic sclerosis with a potential as a disease marker.

OBJECTIVE: Cartilage oligomeric matrix protein (COMP), primarily found in cartilage, is thought to be an important regulator of assembly and maintenance of the fibrillar collagen I and II networks. Recently, COMP was shown to be produced by skin fibroblasts from patients with systemic sclerosis (SSc, or scleroderma). The purpose of this study was to examine whether COMP is released from skin to serum in patients with SSc, and may serve as indicator of activity of skin involvement. METHODS: Serum COMP levels were measured by enzyme linked immunosorbent assay in patients with SSc whose skin involvement was assessed with the modified Rodnan skin score (mRss) and high frequency ultrasound. The presence of COMP in skin biopsies was assessed by Western blot using a monoclonal antibody specific for the very C-terminal end of human COMP. RESULTS: Serum COMP correlated to skin involvement as measured by the mRss (n = 70; r(S) = 0.60; p<0.001), to skin thickness measured by ultrasound (n = 88; r(S) = 0.55; p<0.001) and inversely to skin echogenicity measured by ultrasound (n = 88; r(S) = -0.40; p<0.001). In 70 patients followed longitudinally there was a correlation between changes in serum COMP (n = 307) and changes in mRss (r(S) = 0.35; p = 0.008). In individual patients monitored with repeated measurements, serum COMP changes closely paralleled changes in mRss. A C-terminal COMP fragment, with an apparent molecular mass of 56 kDa, was identified in SSc skin biopsies, while no COMP reactivity was detected in normal skin. CONCLUSION: The high turnover of COMP in SSc skin suggests a pathophysiological role. Serum COMP shows promise as a new biomarker in SSc.

Breath-Hold Blood Oxygen Level-Dependent MRI: A Tool for the Assessment of Cerebrovascular Reserve in Children with Moyamoya Disease.

BACKGROUND AND PURPOSE: There is a critical need for a reliable and clinically feasible imaging technique that can enable prognostication and selection for revascularization surgery in children with Moyamoya disease. Blood oxygen level-dependent MR imaging assessment of cerebrovascular reactivity, using voluntary breath-hold hypercapnic challenge, is one such simple technique. However, its repeatability and reliability in children with Moyamoya disease are unknown. The current study sought to address this limitation. MATERIALS AND METHODS: Children with Moyamoya disease underwent dual breath-hold hypercapnic challenge blood oxygen level-dependent MR imaging of cerebrovascular reactivity in the same MR imaging session. Within-day, within-subject repeatability of cerebrovascular reactivity estimates, derived from the blood oxygen level-dependent signal, was computed. Estimates were associated with demographics and intellectual function. Interrater reliability of a qualitative and clinically applicable scoring scheme was assessed. RESULTS: Twenty children (11 males; 12.1 ± 3.3 years) with 30 MR imaging sessions (60 MR imaging scans) were included. Repeatability was "good" on the basis of the intraclass correlation coefficient (0.70 ± 0.19). Agreement of qualitative scores was "substantial" (κ = 0.711), and intrarater reliability of scores was "almost perfect" (κ = 0.83 and 1). Younger participants exhibited lower repeatability (P = .027). Repeatability was not associated with cognitive function (P > .05). However, abnormal cerebrovascular reactivity was associated with slower processing speed (P = .015). CONCLUSIONS: Breath-hold hypercapnic challenge blood oxygen level-dependent MR imaging is a repeatable technique for the assessment of cerebrovascular reactivity in children with Moyamoya disease and is reliably interpretable for use in clinical practice. Standardization of such protocols will allow further research into its application for the assessment of ischemic risk in childhood cerebrovascular disease.

The time course of natriuretic hormones as plasma markers of myocardial recovery in heart transplant candidates during ventricular assist device support reveals differences among device types.

BACKGROUND: The natriuretic hormones ANP and BNP are expressed differently in the myocardium. Both hormones have compensatory diuretic activity during heart failure. Mechanical stretch of the myocardial walls induces the expression of these hormones. In failing human myocardium, both ANP and BNP are transcribed in the ventricular myocardium in high amounts. We measured the plasma concentrations of ANP and BNP in patients supported by various ventricular assist devices (VADs) at various times. We analyzed the time courses of ANP and BNP to determine (1) the time scale of their down-regulation as a marker of putative myocardial recovery, (2) their steady-state levels under VAD support and (3) differences caused by various VAD devices. METHODS: We analyzed ANP and BNP using commercially available radioimmune assays. We analyzed the time courses of patients supported by Thoratec (THO) LVAD (n = 8), TCI Heartmate (TCI) (n = 6), Novacor (NOV) (n = 7), and Lionheart (LIO) (n = 3). RESULTS: Patients supported with NOV and some patients with TCI showed down-regulation of BNP to a steady-state level at 30 to 50 days, following a single exponential decay. In contrast, patients supported by THO or LIO did not reveal a determined time course of the natriuretic hormones. Only a few patients reached normal plasma values during VAD support. CONCLUSION: The time courses of ANP and BNP differ among VAD types because of construction and/or driving mode, which might be important when considering patients for weaning from VAD without heart transplant.

Combined fat- and water-suppressed MR imaging of orbital tumors.

BACKGROUND AND PURPOSE: The use of a high-resolution T2-weighted MR sequence, which suppresses signal from both fat and water, has been shown to be highly effective for depicting areas of inflammatory damage within the optic nerve. The ability of this sequence to show neoplastic and inflammatory orbital lesions, which may mimic neuritis, is unknown. This study was designed to examine the characteristics of such a sequence for the investigation of orbital mass lesions. METHODS: Twenty-eight patients with known or suspected mass lesions of the orbit and six healthy volunteers were recruited for study. Imaging was performed with a 1.5-T MR unit. Participants were examined by selective partial inversion recovery (SPIR) sequences with T2-weighted fast spin-echo acquisition, selective partial inversion recovery/fluid attenuated inversion recovery (SPIR/FLAIR) sequences with fast spin-echo acquisition, short tau inversion recovery (STIR) sequences with fast spin-echo acquisition, and SPIR sequences with contrast-enhanced T1-weighted fast spin-echo acquisition. Two neuroradiologists, using a randomised, blinded method, scored images for lesion presence and extent. Lesion extent was defined as the number of images with visible abnormality, and was compared with the standard of reference established at a later date by consensus review of all imaging sequences. The ability of the sequences to show the presence and extent of pathologic lesions was compared. RESULTS: The SPIR/FLAIR sequence showed both the presence and extent of orbital masses significantly better than did either STIR or T2-weighted SPIR sequences (P<.01 and P<.001, respectively). Contrast-enhanced T1-weighted SPIR images ranked better than SPIR/FLAIR images, although the difference failed to reach statistical significance. In the orbital apex, the SPIR/FLAIR technique was superior to all other techniques used. This reflected its ability to distinguish enhancing, pathologic lesions from enhancing, normal anatomy. CONCLUSION: SPIR/FLAIR is an appropriate screening technique for orbital masses and offers significant advantages over currently used fat-suppressed sequences for the investigation of orbital disease.

Comparison of three-dimensional visualization techniques for depicting the scala vestibuli and scala tympani of the cochlea by using high-resolution MR imaging.

BACKGROUND AND PURPOSE: Cochlear implantation requires introduction of a stimulating electrode array into the scala vestibuli or scala tympani. Although these structures can be separately identified on many high-resolution scans, it is often difficult to ascertain whether these channels are patent throughout their length. The aim of this study was to determine whether an optimized combination of an imaging protocol and a visualization technique allows routine 3D rendering of the scala vestibuli and scala tympani. METHODS: A submillimeter T2 fast spin-echo imaging sequence was designed to optimize the performance of 3D visualization methods. The spatial resolution was determined experimentally using primary images and 3D surface and volume renderings from eight healthy subjects. These data were used to develop the imaging sequence and to compare the quality and signal-to-noise dependency of four data visualization algorithms: maximum intensity projection, ray casting with transparent voxels, ray casting with opaque voxels, and isosurface rendering. The ability of these methods to produce 3D renderings of the scala tympani and scala vestibuli was also examined. The imaging technique was used in five patients with sensorineural deafness. RESULTS: Visualization techniques produced optimal results in combination with an isotropic volume imaging sequence. Clinicians preferred the isosurface-rendered images to other 3D visualizations. Both isosurface and ray casting displayed the scala vestibuli and scala tympani throughout their length. Abnormalities were shown in three patients, and in one of these, a focal occlusion of the scala tympani was confirmed at surgery. CONCLUSION: Three-dimensional images of the scala vestibuli and scala tympani can be routinely produced. The combination of an MR sequence optimized for use with isosurface rendering or ray-casting algorithms can produce 3D images with greater spatial resolution and anatomic detail than has been possible previously.

Effects of dietary fatty acids on the composition and oxidizability of low-density lipoprotein.

OBJECTIVE: The objective of this study was to compare the effects of dietary monounsaturated fatty acids (MUFA), n-6 and n-3 polyunsaturated fatty acids (PUFA) on LDL composition and oxidizability. DESIGN, SETTING AND SUBJECTS: Sixty-nine healthy young volunteers, students at a nearby college, were included. Six subjects withdrew because of intercurrent illness and five withdrew because they were unable to comply with the dietary regimen. INTERVENTIONS: The participants received a 2-week wash-in diet rich in saturated fatty acids (SFA) followed by diets rich in refined olive oil, rapeseed oil or sunflower oil for 4 weeks. Intakes of vitamin E and other antioxidants did not differ significantly between the diets. RESULTS: At the end of the study, LDL oxidizability was lowest in the olive oil group (lag time: 72.6 min), intermediate in the rapeseed oil group (68.2 min) and highest in the sunflower oil group (60.4 min, P<0.05 for comparison of all three groups). Despite wide variations in SFA intake, the SFA content of LDL was not statistically different between the four diets (25.8-28.5% of LDL fatty acids). By contrast, the PUFA (43.5%-60.5% of LDL fatty acids) and MUFA content of LDL (13.7-29.1% of LDL fatty acids) showed a wider variability dependent on diet. CONCLUSIONS: Enrichment of LDL with MUFA reduces LDL susceptibility to oxidation. As seen on the rapeseed oil diet this effect is independent of a displacement of higher unsaturated fatty acids from LDL. Evidence from this diet also suggests that highly unsaturated n-3 fatty acids in moderate amounts do not increase LDL oxidizability when provided in the context of a diet rich in MUFA.

Electrogenicity of hepatocellular fatty acid uptake.

Sensitivity of cellular fatty acids uptake to the membrane potential difference is still a matter of controversy. For direct evaluation of potential sensitivity the effect of changing membrane potential on uptake of a fluorescent long chain fatty acid derivative, 12-NBD-stearate, in isolated rat hepatocytes, was examined. Changes in membrane potential were achieved by patch clamp procedures. Fatty acid influx was simultaneously determined by recording of cell fluorescence. Hyperpolarization from -30 to -70 mV accelerated fatty acid influx whereas depolarization to +50 mV reduced uptake. After obtaining equilibrium hyperpolarization increased cell fluorescence, whereas depolarization pushed NBD-stearate out of cells. Potential sensitivity of uptake was dependent on the fatty acid concentrations in the medium with most prominent effects at low unbound concentrations. These data show that, at low fatty acid concentrations, uptake is, in part, driven by an intracellular negative electric membrane potential.

Normal-appearing white matter permeability distinguishes poor cognitive performance in processing speed and working memory.

BACKGROUND AND PURPOSE: Secondary-progressive MS is characterized by reduced acute inflammation and contrast enhancement but with increased axonal degeneration and cognitive/clinical disability that worsens with advanced disease. Relative recirculation, extracted from DSC is a surrogate measure of BBB integrity. We hypothesized that normal-appearing white matter relative recirculation is reduced in cognitively impaired compared with nonimpaired secondary-progressive MS, reflecting more advanced disease. MATERIALS AND METHODS: Cognitive performance was classified as impaired or nonimpaired by use of Minimal Assessment of Cognitive Function In MS test components. Demographic data, brain parenchymal fraction, WM lesion fraction, and weighted mean normal-appearing white matter relative recirculation were compared in cognitively dichotomized groups. Univariate and multivariate logistic regressions were used to study the association between cognitive test results and normal-appearing white matter relative recirculation. RESULTS: The mean (SD) age of 36 patients with secondary-progressive MS studied was 55.9 ± 9.3 years; 13 of 36 (36%) patients were male. A highly significant difference between normal-appearing white matter relative recirculation and WM lesion relative recirculation was present for all patients (P < .001). Normal-appearing white matter relative recirculation in impaired patients was significantly lower than in nonimpaired subjects for the Symbol Digit Modalities Test (P = .007), Controlled Word Association Test (P = .008), and Paced Auditory Serial Addition Test (P = .024). The Expanded Disability Status Scale demonstrated an inverse correlation with normal-appearing white matter relative recirculation (r = -0.319, P = .075). After adjustment for confounders, significant normal-appearing white matter relative recirculation reduction persisted for the Symbol Digit Modalities Test (P = .023) and the Paced Auditory Serial Addition Test (P = .047) but not for the Controlled Word Association Test (P = .13) in impaired patients. CONCLUSIONS: Significant normal-appearing white matter relative recirculation reduction exists in cognitively impaired patients with secondary-progressive MS, localizing to the domains of processing speed and working memory.

Wavelets and fuzzy relational classifiers: a novel diffusion-weighted image analysis system for pediatric metabolic brain diseases.

The diffusion-weighted imaging (DWI) technique can be utilized to investigate a variety of diseases. We propose an automated pilot system, which assists in the diagnosis of metabolic brain diseases, utilizing the DWI. In this study, DWI images are preprocessed and exponential apparent diffusion coefficient (eADC) images are produced. The eADC images are later brain extracted and normalized to a standard brain template. Subsequently, we utilized wavelets to denoise the eADC images. The images are rectified, thresholded and now conspicuous abnormal regions are subsequently identified utilizing different brain atlases. Abnormal regions constitute the features that will be used by a fuzzy relational classifier in order to categorize the diseases. A sensitivity and specificity of 60% and 93.33%, respectively, in detecting metabolic brain diseases have been achieved.

Texture analysis: a review of neurologic MR imaging applications.

Texture analysis describes a variety of image-analysis techniques that quantify the variation in surface intensity or patterns, including some that are imperceptible to the human visual system. Texture analysis may be particularly well-suited for lesion segmentation and characterization and for the longitudinal monitoring of disease or recovery. We begin this review by outlining the general procedure for performing texture analysis, identifying some potential pitfalls and strategies for avoiding them. We then provide an overview of some intriguing neuro-MR imaging applications of texture analysis, particularly in the characterization of brain tumors, prediction of seizures in epilepsy, and a host of applications to MS.

Contrast-enhanced MR imaging in acute ischemic stroke: T2* measures of blood-brain barrier permeability and their relationship to T1 estimates and hemorrhagic transformation.

BACKGROUND AND PURPOSE: rtPA is an effective treatment for AIS, yet it is substantially underused due to the increased risk of HT. Recent work suggests that permeability-related information can be extracted from routine T2*-based perfusion images by measuring the rR of the contrast agent. Given that other T2*-based measures have recently been proposed, the purpose of this study was to evaluate 4 such permeability measures in identifying patients with AIS who will proceed to HT. MATERIALS AND METHODS: Eighteen patients with AIS were examined within a mean of 3.3 +/- 1.4 hours postonset. Dynamic T2*-weighted imaging consisted of a single-shot EPI following a bolus of gadodiamide. HT was determined on follow-up CT or MR imaging at 24-72 hours. Mean values of rR, Peak Height, Recovery, as well as Slope were calculated and analyzed on the basis of follow-up HT status. RESULTS: Eight patients proceeded to HT. The mean rR for patients with HT was significantly greater than that for patients without HT (0.22 +/- 0.06 versus 0.14 +/- 0.06, P = .006), while there was a trend toward decreased %Recovery in patients with HT (76 +/- 6 versus 82 +/- 11%, P = .092). There was a significant negative correlation between %Recovery and rR (r = -0.88, P < .001). No significant differences or trends were detected with respect to Peak Height or Slope. CONCLUSIONS: Both rR and %Recovery can be readily extracted from a routine perfusion MR imaging dataset and show potential for identifying HT during the acute phase poststroke.

Recombinant tissue plasminogen activator increases blood-brain barrier disruption in acute ischemic stroke: an MR imaging permeability study.

BACKGROUND AND PURPOSE: Although thrombolytic therapy (recombinant tissue plasminogen activator [rtPA]) represents an important step forward in acute ischemic stroke (AIS) management, there is a clear need to identify high-risk patients. The purpose of this study was to investigate the role of quantitative permeability (KPS) MR imaging in patients with AIS treated with and without rtPA. We hypothesized that rtPA would increase KPS and that KPS MR imaging can be used to predict the risk of hemorrhagic transformation (HT). MATERIALS AND METHODS: Thirty-six patients with AIS were examined within a mean of 3.6 hours of documented symptom onset. KPS MR imaging was performed as part of our AIS protocol. KPS coefficients in the stroke lesion were estimated for all patients, and the relationship between KPS and both HT and rtPA was investigated by using Student t tests. Receiver operating characteristic (ROC) curves were computed for predicting HT from KPS. RESULTS: The occurrence rate of HT for patients who received rtPA and those who did not was 43% and 37%, respectively. Assessment of KPS in the lesion revealed significant differences between those who hemorrhaged and those who did not (P < .0001) as well as between rtPA-treated and untreated patients (P = .008). ROC analysis indicated a KPS threshold of 0.67 mL/100 g/min, with a sensitivity of 92% and a specificity of 78%. CONCLUSIONS: The results of this study indicate that KPS is able to identify patients at higher risk of HT and may allow use of physiologic imaging rather than time from onset of symptoms to guide treatment decision.

PCA-SGA implementation in classification and disease specific feature extraction of the brain MRS signals.

The medical diagnostic systems often suffer from the high dimensional data. In this study, Principle Component Analysis (PCA) has been used for dimensionality reduction of the brain Magnetic Resonance Spectroscopy (MRS) signals. Afterwards, the Simple Genetic Algorithms (SGA) is utilized in order to classify different brain diseases. SGA is later used to extract MRS signal features in case of metabolic brain diseases (MD). The PCA-SGA implementation received the specificity of 89.91%. The SGA was able to achieve the sensitivity of 84.84% and positive predictivity of 88.46% in extracting disease specific MRS signal features.

Effect of surface and intracellular pH on hepatocellular fatty acid uptake.

Fatty acids enter hepatocytes, at least in part, by a carrier-mediated uptake mechanism. The importance of driving forces for fatty acid uptake is still controversial. To evaluate possible driving mechanisms for fatty acid transport across plasma membranes, we examined the role of transmembrane proton gradients on fatty acid influx in primary cultured rat hepatocytes. After hepatocytes were loaded with SNARF-1 acetoxymethyl ester, changes in intracellular pH (pHi) under different experimental conditions were measured and recorded by confocal laser scanning microscopy. Fatty acid transport was increased by 45% during cellular alkalosis, achieved by adding 20 mM NH4Cl to the medium, and a concomitant paracellular acidification was observed. Fatty acid uptake was decreased by 30% during cellular acidosis after withdrawal of NH4Cl from the medium. Cellular acidosis activates the Na+/H+ antiporter to export excessive protons to the outer cell surface. Inhibition of Na+/H+ antiporter activity by amiloride diminishes pHi recovery and thereby accumulation of protons at the outer surface of the plasma membrane. Under these conditions, fatty acid uptake was further inhibited by 57% of control conditions. This suggests stimulation of fatty acid influx by an inwardly directed proton gradient. The accelerating effect of protons at the outer surface of the plasma membrane was confirmed by studies in which pH of the medium was varied at constant pHi. Significantly higher fatty acid influx rates were observed at low buffer pH. Recorded differences in fatty acid uptake appeared to be independent of changes in membrane potential, because BaCl2 did not influence initial uptake velocity during cellular alkalosis and paracellular acidosis. Moreover, addition of oleate-albumin mixtures to the NH4Cl incubation buffer did not change the observed intracellular alkalinization. In contrast, after cells were acid loaded, addition of oleate-albumin solutions to the recovery buffer increased pHi recovery rates from 0.21 +/- 0.02 to 0.36 +/- 0.05 pH units/min (P < 0.05), indicating that fatty acids further stimulate Na+/H+ antiporter activity during pHi recovery from an acid load. It is concluded that carrier-mediated uptake of fatty acids in hepatocytes follows an inwardly directed transmembrane proton gradient and is stimulated by the presence of H+ at the outer surface of the plasma membrane.

Sodium, hydrogen antiporter activation by extracellular adenosine triphosphate in biliary epithelial cells.

BACKGROUND & AIMS: Extracellular nucleotides are secretagogues and influence ion permeability. The aim of this study was to investigate whether secretagogues activate transmembrane acid-base carriers, e.g., sodium, hydrogen antiporter in cholangiocytes. METHODS: Cells were loaded with pH and Ca(2+)-sensitive dyes. Intracellular changes in ion concentrations were monitored by confocal laser scanning microscopy. Adenosine 3', 5'-cyclic monophosphate was determined by standard methods. RESULTS: Baseline intracellular pH (pH1) averaged 7.28 +/- 0.17 pH units. Adenosine triphosphate (ATP; 10 mumol/L) increased baseline pH1 by 0.28 +/- 0.06 pH units/10 min (P < 0.01). Ten and 100 mumol/L ATP increased Na+, H+ antiporter-mediated proton flux from 9.4 +/- 4.9 mmol. L-1 min-1 to 17.2 +/- 11.8 and 16.7 +/- 10.3 mmol. L(-1)-min(-1) (P < 0.001), respectively. Under control and ATP-stimulated conditions, 1 mmol/L amiloride blocked pH1 recovery, indicating true activation of Na+, H+ antiporter by extracellular ATP. Inhibition of basolateral Na+, H+ antiporter isoform inhibited stimulation by ATP. Na+, H+ antiporter-mediated proton flux was stimulated by adenosine, uridine triphosphate, adenosine-5'-0-(3-thiotriphosphate), alpha, beta-methylene-ATP, and 2-methylthio-ATP but not prevented by adenosine receptor blocking. Activation by ATP was not influenced by the Ca2+/protein kinase C/calmodulin system but could be mimicked by addition of N6,2'-O-dibutyryladenosine-3',5'-cyclic monophosphate and was inhibited by pertussis toxin. CONCLUSIONS: Extracellular nucleotides may modulate secretory and absorptive function of cholangiocytes by activating Na+/H+ exchange mechanisms.

Anatomical variation of cerebral venous drainage: the theoretical effect on jugular bulb blood samples.

Recent studies have demonstrated significant variation in bilateral jugular venous oxygen saturation measurements which may be of clinical significance. We have therefore measured variations in normal dural sinus venous drainage to assess the possible effects of normal anatomical variations on measured jugular venous oxygen saturation. Normal volunteers (n = 25) were imaged using magnetic resonance venography to demonstrate variations in venous anatomy. Flow was measured in the superior sagittal sinus and bilaterally in the transverse sinus, sigmoid sinus proximal to the jugular bulb and proximal jugular vein using phase difference magnetic resonance imaging. Examination of magnetic resonance venogram images showed considerable variability in the symmetry of transverse sinus flow. Complete absence of one transverse sinus was seen in four cases and significant asymmetry in the size of the transverse sinuses was present in 13. Quantitative flow studies demonstrated that the ratio of superior sagittal sinus to combined jugular bulb flow showed remarkably little variation (0.46 +/- 0.06). Measurements of transverse sinus flow showed significant asymmetry (< 40% of superior sagittal sinus flow in one transverse sinus) in 21 of 25 volunteers. The effect of the observed asymmetry on jugular venous oxygen saturation was modelled based on the assumption of either a supratentorial or infratentorial lesion. This model predicted significant asymmetry in jugular venous oxygen saturation measurements (> 10%) in 65% of cases with a supratentorial lesion which is in close agreement with clinical observations. This study suggests that normal variations in venous drainage may account for observed asymmetry in jugular venous oxygen saturation measurements.

Degradation of the cyclic AMP antagonist prostaglandylinositol cyclic phosphate (cyclic PIP) by dephosphorylation.

The cAMP antagonist, prostaglandylinositol cyclic phosphate (cyclic PIP), is synthesized from prostaglandin E and activated inositol phosphate. From various tissues only that amount of cyclic PIP can be isolated that constitutes the difference between synthesis and degradation. In order to overcome this drawback, the cyclic PIP degrading enzyme or enzymes had to be characterized prior to searching for inhibitors. Cyclic PIP degrading activities have been found in all rat tissues tested, and are lowest in brain (380 pmol x min(-1) x g(-1) wet weight) and highest in liver (1460 pmol x min(-1) x g(-1) wet weight). They are associated primarily with particulate structures of the cells, but not with the plasma membrane. There appear to be at least two different enzymatic activities involved in the degradation of cyclic PIP, because there are two pH-optima, one between pH 7 and 8 and another between pH 4 and 5. It is assumed that these activities are located in microsomes and lysosomes. Because prostaglandylinositol is the final product obtained in the degradation of cyclic PIP, a phosphodiesterase and a phosphatase should be involved, which could not yet be identified individually. Like alkaline phosphatase, cyclic PIP-degrading enzymes require Mg2+ and they are inhibited by heavy metal ions such as mercuric and copper chloride, by sodium fluoride and interestingly, by prostaglandins.

Reproducibility of T2* blood volume and vascular tortuosity maps in cerebral gliomas.

The development of anti-angiogenic therapies for tumors has led to a demand for imaging-based surrogate markers of the angiogenic process. The utility of such markers is highly dependent on their test-retest reproducibility. This paper presents a formal assessment of the reproducibility of measurements of relative blood volume (rBV), normalized rBV (rBVnorm), and vascular tortuosity as estimated by measurement of relative recirculation (rR). The study was conducted in 11 patients with glioma who were scanned on two occasions 36-56 hours apart. The observed reliability estimates were used to calculate 95% confidence limits for detection of differences between groups and for changes in individual cases. The results show that measurement of rBV or rBVnorm in consecutive studies is statistically capable of reliably detecting changes in excess of 15% in between group studies and 25% in individual patients. Measurement of vascular tortuosity using is less reproducible but is able to confidently identify changes in excess of 30% in group studies and 35% in individuals.

Absence of health insurance is associated with decreased life expectancy in patients with cystic fibrosis.

Life expectancy for individuals with cystic fibrosis (CF) has increased dramatically in the last 30 yr, but it is unclear whether the improved survival has applied equally to individuals with different health insurance status. We developed a retrospective inception cohort of all 189 patients with CF born 1/1/55 to 12/31/70 who had at least one hospitalization at a university referral center. The median survival for patients with CF who were without health insurance was 6.1 yr compared with 20.5 yr for those with Medicaid and 20.5 yr for those with private insurance. Using multivariate Cox regression, health insurance and increased socioeconomic status were independently associated with longer survival. The adjusted relative risk of death was greater for the absence of health insurance than for factors previously shown to predict mortality in individuals with CF (female sex and presentation with meconium ileus). In summary, the absence of health insurance was associated with increased mortality rate in children with CF and was a stronger predictor of mortality than variables previously shown to be associated with mortality for CF. If increasing numbers of children with CF lose health insurance coverage, our results suggest that their life expectancy will decrease dramatically.

Muscarinic acetylcholine receptor stimulation of biliary epithelial cells and its effect on bile secretion in the isolated perfused liver [corrected].

The aim was to explore whether biliary epithelial cells show muscarinic acetylcholine receptors and to investigate their role in ductular bile formation. In both, isolated rat biliary epithelial cells and Mz-Cha-1 cells, a biliary epithelial cell line, binding of [3H]N-methyl-scopolamine occurred with 0.718 +/- 0.08 and 0.482 +/- 0.05 fmol per 10(6) cells, respectively. To characterize the involved second messenger, intracellular Ca2+ levels were monitored by confocal microscopy. Stimulation of biliary epithelial cells with carbachol produced an increase in free cytosolic Ca2+ levels that declined to baseline values describing a sinusoidal oscillation curve. Increasing concentrations of the agonist decreased latency of the response and increased oscillation frequency. Similar results were obtained in Mz-Cha-1 cells. The intracellular Ca2+ originated from IP3 sensitive intracellular stores and from the extracellular medium. The Ca2+ response could partially be blocked by atropine and completely by pirenzepine, a specific muscarinic receptor-type M1 antagonist. The presence of M1 receptor messenger RNA (mRNA) in biliary epithelial cells was confirmed by reverse transcriptase polymerase chain reaction. In the isolated perfused guinea pig liver, a model with high ductular bile flow, carbachol induced a dose dependent decrease of bile flow by 79.6% +/- 9.8% at 50 mumol/L carbachol (P < .001), without affecting perfusion pressure or biliary electrolyte concentrations. It is concluded that biliary epithelial cells express muscarinic acetylcholine receptors. Stimulation of this receptor leads to cholestasis. This could be because of changes in peribiliary permeability and/or inhibition of biliary epithelial cell secretory function.