Validation of the Capsule Endoscopy Crohn's Disease Activity Index (CECDAI or Niv score): a multicenter prospective study.

BACKGROUND AND STUDY AIMS: The Capsule Endoscopy Crohn's Disease Activity Index (CECDAI or Niv score) was devised to measure mucosal disease activity using video capsule endoscopy (VCE). The aim of the current study was to prospectively validate the use of the scoring system in daily practice. METHODS: This was a multicenter, double-blind, prospective, controlled study of VCE videos from 62 consecutive patients with isolated small-bowel Crohn's disease. The CECDAI was designed to evaluate three main parameters of Crohn's disease: inflammation (A), extent of disease (B), and stricture (C), in both the proximal and distal segments of the small bowel. The final score was calculated by adding the two segmental scores: CECDAI = ([A1 × B1] + C1) + ([A2 × B2] + C2). Each examiner in every site interpreted 6 - 10 videos and calculated the CECDAI. The de-identified CD-ROMs were then coded and sent to the principal investigator for CECDAI calculation. RESULTS: The cecum was reached in 72 % and 86 % of examinations, and proximal small-bowel involvement was found in 56 % and 62 % of the patients, according to the site investigators and principal investigator, respectively. Significant correlation was demonstrated between the calculation of the CECDAI by the individual site investigators and that performed by the principal investigator. Overall correlation between endoscopists from the different study centers was good, with r = 0.767 (range 0.717 - 0.985; Kappa 0.66; P < 0.001). There was no correlation between the CECDAI and the Crohn's Disease Activity Index or the Inflammatory Bowel Disease Quality of Life Questionnaire or any of their components. CONCLUSION: A new scoring system of mucosal injury in Crohn's disease of the small intestine, the CECDAI, was validated. Its use in controlled trials and/or regular follow-up of these patients is advocated.

Influence of calcium on phosphorylation of a synaptosomal protein.

Synaptosomal proteins isolated from rat cerebral cortex were phosphorylated endogeneously in the presence of [gamma-32P]ATP. The phosphorylated proteins were found to be membrane bound by differential and density gradient centrifugation. In contrast to the phosphorylation of all synaptosomal proteins, phosphorylation of one protein (C), 41 000--43 000 daltons, was inhibited by Mg2+ and stimulated by Ca2+. In addition, the ionophores X537A and A23187, as well as papaverine, selectively enhanced phosphorylation of protein C without affecting phosphorylation of the outer proteins. Cyclic AMP did not influence the phosphorylation of protein C but markedly affected the phosphorylation of other synaptosomal proteins. It appears that the phosphorylation of protein C is stimulated by agents which trigger the release of neurotransmitters (Ca2+, X537A, A23187 and papaverine), and is inhibited by Mg2+, which inhibits release. It is proposed that the phosphorylation of protein C is related to membranal events underlying the release of neurotransmitters.

The characterization and phosphorylation of an actin-like protein in synaptosomal membranes.

A protein of 43,000 daltons, named protein 'C', is a component of synaptosomal plasma membranes, vesicular and microsomal membranes, as well as synaptosomal and cellular cytoplasm. Protein 'C' undergoes endogeneous phosphorylation in synaptosomal plasma membranes but not in other subcellular fractions. This phosphorylation is stimulated by papaverine and calcium, inhibited by magnesium and not affected by cyclic nucleotides. Protein 'C' and muscle actin were shown to be very similar by isoelectric focusing, two dimensional gel electrophoresis, and by peptide mapping. This suggests that protein 'C' is an actin-like protein which undergoes endogenous phosphorylation specifically in synaptosomal plasma membranes. Phosphorylation of protein 'C' may be involved in neurotransmitter release.

The effect of ACTH on rat brain synaptic plasma membrane lipid fluidity.

The effect of ACTH on the lipid fluidity was examined in synaptic plasma membranes from rat forebrain. ACTH1-24 increased the fluidity of the synaptic plasma membranes in a dose-dependent way, the lowest effective dose being 10(-5) M. The shorter N-terminal fragment ACTH1-10 was not effective. The significance of this finding is discussed in relation to the known effects of ACTH on synaptic membrane phosphorylation.

Binding of platelet-activating factor to platelets of Alzheimer's disease and multiinfarct dementia patients.

The binding of 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine (platelet-activating factor, PAF) to platelets was studied in 22 patients with probable Alzheimer's disease (AD), 11 with multi-infarct dementia (MID), 22 age-matched normal old controls, and 20 young subjects. The results showed a significantly lower degree of PAF binding to platelets of AD and MID patients than in those of the old controls and young subjects (133.3 +/- 8.5, and 123.4 +/- 16.5 vs. 202.3 +/- 11.6 and 206.7 +/- 17.3 receptors/cell, respectively; p < 0.01). These differences were due to reduced Bmax, while Kd remained unchanged. No significant difference was observed between the PAF binding to platelets of AD and MID patients nor between that of old and young controls. No correlation was found between age and binding in the various elderly groups. However, a significant correlation was found between PAF binding and degree of cognitive impairment in the AD patients. This is the first evidence to support a possible involvement of PAF in dementing disorders.

Increased cytosolic free calcium in lymphocytes of Alzheimer patients.

Free cytosolic calcium content [Ca2+]i was determined in peripheral blood mononuclear cells (PBMC) from healthy volunteers, Alzheimer's disease and multi-infarct dementia patients. Measurement of [Ca2+]i by the fluorescent dye quin-2, before and at several time intervals during incubation with phytohemagglutinin (PHA), showed a higher resting [Ca2+]i in PBMC of Alzheimer's disease patients as compared to controls and multi-infarct dementia patients. However, the addition of supra-optimal PHA doses (100 micrograms/ml) induced strikingly higher [Ca2+]i levels in Alzheimer's disease patients (1647 +/- 200 nM versus 398 +/- 27 nM in controls, and 346 +/- 40 nM in multi-infarct dementia patients). The increased [Ca2+]i concentration was also found after a specific stimulation with a monoclonal anti-CD3 antibody. The results may have important implications in understanding the pathophysiology of Alzheimer's disease and suggest that [Ca2+]i may prove diagnostically valuable.

Alzheimer's dementia and binding to alpha 2 adrenoreceptors in platelets.

Seventy-five patients with probable Alzheimer's disease were screened for binding of alpha 2 receptors (A2R) to their platelet membranes; the results were compared with 51 age- and sex-matched controls. Receptor binding assays were performed using [3H] Yohimbine as the radioligand. The results showed a higher binding capacity in the demented population as compared to the control group (2.18 +/- 0.15 fmol/mg protein, as compared to 1.73 +/- 0.13, P less than 0.03). This increased binding to platelets in the demented patients was more prominent in demented females: 34% higher binding as compared with female controls (2.06 +/- 0.5 vs 1.54 +/- 0.04). The difference between demented and normal males was less (2.34 +/- 0.05 vs 1.88 +/- 0.05). The results indicate an involvement of the A2R system, either primarily or secondarily, in the disease process. Since there is an overlap between results from the patients with Alzheimer's disease and the normal subjects, A2R may serve as only a supportive marker for Alzheimer's disease.

The effect of chronic diazepam treatment on discrimination performance and 3H-flunitrazepam binding in the brains of shocked and nonshocked rats.

The purpose of the present study was: (1) to investigate the effects of unavoidable shock on an appetitively motivated discrimination task; (2) to evaluate the effect of chronic diazepam treatment on the performance of a previously learned discrimination task in shocked and nonshocked animals; (3) to measure the binding of 3H-flunitrazepam (an analogue of diazepam) to selected brain regions of chronically diazepam-treated shocked and nonshocked rats, in comparison to saline-treated controls. Results indicated that unavoidable shock significantly interfered with the learning of a new, nonshock-related discrimination task. The effect of chronic diazepam treatment on the performance depended on the previous experience of the animal; chronic diazepam treatment significantly improved the maze performance of shocked animals. On the other hand, chronic diazepam treatment in the nonshocked animals tended to interfere with the performance of the discrimination task. Neurochemical data showed significant reduction in 3H-flunitrazepam binding to diazepam receptors in membranes from the brains of a nonshocked diazepam-treated (CD) group in comparison to a nonshocked saline-treated (CS) group. In contrast, the unavoidable shock-treated diazepam group (SD) showed opposite effects, the binding of 3H-flunitrazepam increasing significantly. A significant increase in the maximal binding sites in the frontal cortex from shocked rats treated with diazepam, compared to the nonshocked diazepam-treated rats, was detected by Scatchard analysis.

Binding of platelet-activating factor to platelets of ischemic stroke patients.

Platelet-activating factor (PAF) has been suggested to play an important role in brain ischemia and stroke. We examined whether binding of PAF to platelets of patients suffering from ischemic stroke differs in the acute and subacute stages, and correlated the results with the severity of neurological presentation. A total of 30 patients with first ever ischemic stroke was studied. The patients were divided into two groups according to stroke severity as assessed by the Scandinavian Stroke Scale (SSS). Binding of PAF to platelets was performed on admission (first determination) and 3 weeks later (second determination) by labeling with [H]PAF. Results of PAF binding were obtained also from 21 healthy age- and sex-matched volunteers. Binding results shortly after stroke onset were significantly lower in the severely affected group (analysis of variance, F = 11.0, p < .001). Moreover, only in these patients was there a change in PAF binding between the first and the sec-nd determinations (Wilcoxon signed ranks test, p = .018). A significant correlation was found between PAF binding and severity of neurologic deficits as assessed by the SSS (correlation coefficient = .52, p = .01). This result shows evidence of PAF system involvement in stroke patients. PAF binding to platelets clearly differs in the acute and subacute stages, and the reduction in the number of binding sites correlates with severity of neurologic deficits. We conclude that PAF binding in platelets may serve as an additional marker of stroke severity.

Basal and activated intracellular calcium ion concentrations in mononuclear cells of Alzheimer's disease and unipolar depression.

We present the results of a case control study on the concentration of intracellular calcium [Ca+2]i in peripheral blood mononuclear cells (PBMC) of patients with Alzheimer's disease (AD), major affective disorder unipolar type (DEP), and elderly controls. We used the fluorescent dye Quin-2 to measure intracellular calcium concentration in PBMC both in their basal resting state [Ca+2]b and after activation [Ca+2]a with phytohemagglutinin (PHA). The [Ca+2]b and [Ca+2]a values in PBMC of AD patients were significantly higher than those of the elderly controls and of the DEP patients. In the DEP patients the [Ca+2]a values in PBMC were significantly higher than those of the elderly controls, but the [Ca+2]b values were not. [Ca+2]i levels did not correlate with age, gender, or duration and severity of the diseases investigated. These findings indicate that higher values of [Ca+2]b and [Ca+2]a may differentiate most AD patients from elderly controls or DEP patients.

The effects of postnatal anoxia on behaviour and on the muscarinic and beta-adrenergic receptors in the hippocampus of the developing rat.

Exposure of rats to 25 min anoxia within 24 h following birth caused behavioural as well as biochemical changes during their development and maturity. Following postnatal anoxia, a significant increase in the concentration of the cholinergic muscarinic receptors in the hippocampus was noted at the early stages of development, between 6 and 20 days of age, but reached normal values at 40 days of age. However, at this age, significant increase in the concentration of beta-adrenergic receptors in the hippocampus was found, which remained significantly high during maturity and adulthood, as compared to controls. Rats submitted postnatally to anoxia exhibited hyperactivity in the open field which was maximal at 20-25 days of age and declined towards normal values at 40 days of age. At maturity, between 60 and 80 days of age, these rats showed poor performance in a complex 6-choice discrimination learning but not in simple differential conditioning. Possible correlations between the behavioural and biochemical findings are discussed.

The effects of dibutyryl cyclic AMP, theophylline and papaverine on the release of 3H-catecholamines from rat brain striatal and cortical synaptosomes.

Papaverine enhances the spontaneous release of 3H-dopamine from rat brain striatal synaptosomes and 3H-noradrenaline from cortical synaptosomes in a dose-dependent way. Neither dibutyryl cyclic AMP nor theophylline had any significant effect on either spontaneous or on potassium-evoked 3H-catecholamine release. It is suggested that the effect of papaverine on catecholamine release is not due to its phosphodiesterase inhibitory potency.

Pretreatment with delta 1-tetrahydrocannabinol and psychoactive drugs: effects on uptake of biogenic amines and on behavior.

Injection of delta 1-tetrahydrocannabinol (THC) into mice increased the uptake into brain synaptosomes of radioactive DA, NE, 5-HT and GABA, with the effect on GABA being the greatest; uptake of leucine was not stimulated, indicating that THC does not facilitate the transport of amino acids in general. The effect of THC was stereospecific because pretreatment with the non-psychoactive isomer, (+) delta 6-THC had no effect on uptake of DA into cortical synaptosomes. The effect on DA uptake was correlated with psychoactive potency. THC enhanced uptake more than did SP-111 (a water soluble ester of THC but less potent than THC) and much more than cannabidiol (a non-psychoactive ingredient of marijuana). THC increased the uptake of DA into striatal synaptosomes but much less than into cortical synaptosomes. The enhancing effect of THC on uptake in cortex showed tolerance after chronic (1 week) treatment with THC. Catecholaminergic receptor antagonists (chlorpromazine, propranolol), like THC, stimulated the uptake of DA or NE into cortical synaptosomer. In contrast, pretreatment with MAO (pargyline) or uptake (tricyclic antidepressants) inhibitors, or with amphetamine, decreased uptake. Thus THC does not inhibit MAO uptake, or stimulate the release of catecholamines but may interact with a receptor. The notion of a THC--receptor interaction is supported by behavioral experiments.

Sexual behavior decreases pain sensitivity and stimulated endogenous opioids in male rats.

In male rats copulation has antinociceptive effects as measured either by shock-induced vocalizations or hindlimb withdrawal to pinch. Prolonged mating reduces the content of endogenous opioids in midbrain but not in hypothalamus or caudate nucleus. Blockage of opiate receptors with the narcotic antagonist naloxone (4 mg/kg) significantly extends the postejaculatory interval. The results indicate that mating is a biological stimulus for the release of endogenous opoids, possibly to (a) prevent intense sexual stimulation from becoming aversive, and (b) increase its reward value.

The muscarinic cholinergic receptors in the posterior hypothalamus of hypertensive and normotensive rats.

The density of [3H]quinuclidin-3-yl benzilate ([3H]QNB) binding sites in the posterior hypothalamus was determined in spontaneously hypertensive (SHR) and Wistar Kyoto (WKY) rats at the ages of 1, 3, 11 and 50 weeks. In SHR, even at the early age of 1 week which is prehypertensive, the values obtained were 1.5 times greater than those of age-matched WKY rats. The values of the equilibrium dissociation constant (KD) did not differ between SHR and WKY rats of the same age. In the pons medulla, however, the density of [3H]QNB binding sites was not different between the two strains of rats of matching age. Isolation-induced hypertension in adult Wistar rats and an increase in the density of [3H]QNB binding sites in the posterior hypothalamus were observed to arise concomitantly. A hypothesis is offered whereby a relative increase in ACh receptor sites in the posterior hypothalamus is a primary cause of hypertension in the models considered.

Lipid-induced modulation of opiate receptors in mouse brain membranes.

The binding of [3H] D-Ala-enkephalinamide (DAEA) to crude mitochondrial fractions (P2M) from mouse forebrain was determined after modulation of membrane lipid microviscosity. Lipid fluidization of P2M membranes, following treatment with egg lecithin, resulted in a 50% loss of specific binding of DAEA. Increasing the P2M lipid microviscosity, by incorporation of cholesteryl hemisuccinate (CHS), increased the accessibility of the opiate receptors up to a peak level of 170% which decreased sharply upon further increase in lipid microviscosity. The processes resulting from lipid rigidification may have important implications for aging and for drug addiction.

Idiopathic retroperitoneal fibrosis: implications for a systemic disorder.

A thirty-nine year old woman with idiopathic retroperitoneal fibrosis (IRF) had symptoms, physical abnormalities, and abnormal laboratory tests which supported the hypothesis that IRF is a widespread multisystem disease rather than a localized anatomic disorder.

Infarct volume, neurological severity and PAF binding to platelets of patients with acute cerebral ischemic stroke.

Studies show that Platelet Activating Factor (PAF) is involved in the cerebrovascular response to ischemia, and that its binding to platelets may change in stroke victims. The purpose of this study was to determine whether binding of PAF to platelets of stroke patients could serve as an index for determining the volume of ischemic strokes and severity of neurological presentation. Thirteen stroke patients and 21 healthy controls were studied. The neurological severity of these stroke patients was evaluated by the Scandinavian Stroke Scale. Infarct volume was assessed by planimetric measures of brain CT. PAF binding to platelets was determined by use of radiolabelled PAF. (3H)PAF binding to platelets of stroke patients was lower than in controls (149.58 +/- 46.11 and 212.1 +/- 10.3 receptors cell-1, respectively, p < 0.001) and was significantly correlated with infarct volume (r = -0.606, p = 0.014) and with neurological score (r = 0.527, p = 0.032). No correlation was observed between neurological score and infarct volume. The study confirms the involvement of PAF in the pathogenesis of brain ischemia and neuronal damage. It shows that PAF binding to platelets of stroke patients correlates both with the extent of neuronal damage and the associated neurological impairment, and may serve as an additional index in the assessment of stroke severity and clinical outcome of stroke victims.

The effect of intravenous fluid infusion on blood and urine parameters of hydration and on state of consciousness in terminal cancer patients.

This prospective study was undertaken to assess the state of hydration in terminal cancer patients with and without intravenous fluids during the last 48 hours of their lives and to correlate various measures of hydration with their state of consciousness. We examined indicators of hydration in the plasma and urine of 68 consecutive patients for whom data were available at 48 hours or less before death. Thirteen of the patients were being treated with intravenous (IV) fluids. Nearly all of the patients studied were found to be dehydrated, as determined by laboratory measurements. State of consciousness correlated inversely with serum sodium (p < 0.001) and urine osmolality (p < 0.02). Patients receiving intravenous fluids were not better hydrated than those without IV therapy, nor was their state of consciousness improved. In light of these findings, which suggest there is no clinical benefit from intravenous infusions, decisions regarding intravenous fluid therapy during the last hours of life should be guided by the preferences of the dying patient and his family.