The "interleukin 1 receptor antagonist" is a partial agonist of prostaglandin synthesis by human decidual cells

In many systems the interleukin-1 receptor antagonist opposes the effects of interleukin-1 beta. We considered that it might block interleukin-1 beta-stimulated prostaglandin production from human decidual cells. Very high levels of interleukin-1 receptor antagonist (> 1000 pg/ml) had limited inhibitory effects on IL-1 beta-stimulated PGE2 synthesis, and lower levels of antagonist (< 1000 pg/ml) increased the effects of IL-1 beta. Low concentrations of the antagonist alone (1-100 pg/ml) increased basal PGE2 production, whereas higher levels (10-100 ng/ml) had less effect. It seems, therefore, that in human decidua the "antagonist" is more accurately described as a partial agonist. It has been suggested that the IL-1 receptor antagonist could be used to inhibit decidual prostaglandin synthesis and thereby prevent preterm labor, but this report shows that caution should be exercised before using the receptor antagonist.     

Zonal heterogeneity of hepatic injury following shock/resuscitation: relationship of xanthine oxidase activity to localization of neutrophil accumulation and central lobular necrosis

Post-ischemic hepatic injury is characterized by zonal heterogeneity of injury (central lobular necrosis), sinusoidal neutrophil accumulation, and injury generated by reactive oxygen metabolites. We evaluated the role of the heterogeneous distribution of hepatic xanthine oxidase in the generation of neutrophil accumulation and consequent hepatocellular injury in rats subjected to shock [controlled hemorrhagic hypotension (mean arterial pressure = 37.5 + or - 2.5 mmHg for 120 min)], with or without subsequent resuscitation and hemodynamic stabilization, compared with sham-operated rats. Shock/resuscitation produced striking neutrophil accumulation (assayed by esterase histochemistry) in the pericentral sinusoids, associated with centrolobular necrosis. This paralleled the pericentral distribution of xanthine oxidase (determined by histochemical assay of frozen sections) and its release from the liver into the circulation at resuscitation. Pretreatment with allopurinol inhibited hepatic xanthine oxidase activity, neutrophil accumulation, and pericentral hepatocyte necrosis in shock/resuscitation in rats. These findings suggest that reactive oxygen metabolites generated by heterogeneously distributed xanthine oxidase may contribute to the heterogeneity of hepatocellular injury in "ischemic hepatitis."     

Modeling the interaction between propagating cardiac waves and monophasic and biphasic field stimuli: the importance of the induced spatial excitatory response

INTRODUCTION: Biphasic (BP) defibrillation waveforms have been shown to be significantly more efficacious than equivalent monophasic (MP) waveforms. However, when defibrillation fails, it tends to do so first in distal regions of the heart where induced field gradient magnitudes are lowest. We tested the hypothesis that the improved efficacy of BP waveforms results from their enhanced ability to prevent the initiation of new postshock activation fronts behind preexisting wavetails, rather than from any significantly improved ability to terminate preexisting wavefronts. METHODS AND RESULTS: An idealized computer model of a one-dimensional cardiac strand was used to investigate the spatial and temporal interactions between an underlying propagation front (or tail) and uniform MP or BP field stimuli of various intensities. Axial discontinuities from intercellular junctions induced sawtooth patterns of polarization during such field stimuli, enabling the shocks to interact directly with all cells. MP and BP diastolic thresholds were essentially equal. All suprathreshold MP and BP field stimuli successfully terminated preexisting wavefronts by directly depolarizing tissue ahead of those fronts, thus blocking their continued progression. However, the postshock response at the wavetail was significantly dependent on the shape and strength of the administered field. Low-strength MP stimuli induced an all-or-none excitation response across the wavetail, producing a sharp spatial transmembrane voltage gradient from which a new sustained anterogradely propagating wavefront was initiated. In contrast, low-strength BP field stimuli induced a spatially graded excitatory response whose voltage gradient was insufficient to initiate such a wavefront. Higher-strength MP and BP stimuli both produced graded excitatory responses with no subsequent propagation. CONCLUSIONS: Shock-induced spatial "all-or-none" excitatory responses facilitate, and graded excitatory responses prevent, the postshock initiation of new propagating wavefronts. Moreover, BP field stimuli can induce such graded excitatory responses at significantly lower stimulus strengths than otherwise equivalent MP stimuli. Therefore, these results support an alternative "graded excitatory response" mechanism for the improved efficacy of BP over MP field stimuli in low gradient regions.     

Ion binding and permeation at the GABA transporter GAT1

This study addresses the binding of ions and the permeation of substrates during function of the GABA transporter GAT1. GAT1 was expressed in Xenopus oocytes and studied electrophysiologically as well as with [3H]GABA flux; GAT1 was also expressed in mammalian cells and studied with [3H]GABA and [3H]tiagabine binding. Voltage jumps, Na+ and Cl- concentration jumps, and exposure to high-affinity blockers (NO-05-711 and SKF-100330A) all produce capacitive charge movements. Occlusive interactions among these three types of perturbations show that they all measure the same population of charges. The concentration dependences of the charge movements reveal (1) that two Na+ ions interact with the transporter even in the absence of GABA, and (2) that Cl- facilitates the binding of Na+. Comparison between the charge movements and the transport-associated current shows that this initial Na(+)-transporter interaction limits the overall transport rate when [GABA] is saturating. However, two classes of manipulation--treatment with high-affinity uptake blockers and the W68L mutation-"lock" Na+ onto the transporter by slowing or preventing the subsequent events that release the substrates to the intracellular medium. The Na+ substitutes Li+ and Cs+ do not support charge movements, but they can permeate the transporter in an uncoupled manner. Our results (1) support the hypothesis that efficient removal of synaptic transmitter by the GABA transporter GAT1 depends on the previous binding of Na+ and Cl-, and (2) indicate the important role of the conserved putative transmembrane domain 1 in interactions with the permeant substrates.     

Apparent mtDNA heteroplasmy in Alzheimer's disease patients and in normals due to PCR amplification of nucleus-embedded mtDNA pseudogenes

In an unprecedented finding, Davis et al. [Davis, R. E., Miller, S., Herrnstadt, C., Ghosh, S. S., Fahy, E., Shinobu, L. A., Galasko, D., Thal, L. J., Beal, M. F., Howell, N. & Parker, W. D., Jr. (1997) Proc. Natl. Acad. Sci. USA 94, 4526-4531] used an unusual DNA isolation method to show that healthy adults harbor a specific population of mutated mitochondrial cytochrome c oxidase (COX) genes that coexist with normal mtDNAs. They reported that this heteroplasmic population was present at a level of 10-15% in the blood of normal individuals and at a significantly higher level (20-30%) in patients with sporadic Alzheimer's disease. We provide compelling evidence that the DNA isolation method employed resulted in the coamplification of authentic mtDNA-encoded COX genes together with highly similar COX-like sequences embedded in nuclear DNA ("mtDNA pseudogenes"). We conclude that the observed heteroplasmy is an artifact.     

Genetic defects at the UGT1 locus associated with Crigler-Najjar type I disease, including a prenatal diagnosis

OBJECTIVE: To examine the diaschisis of faultless conventional neuropsychological tests of unilateral spatial neglect (USN) and defects of activities of daily living (ADL) functions. DESIGN: The evaluations of USN by assessments now in wide use were compared with newly developed assessments using a dual-task methodology in which a stepping activity was used as a "distracter" during USN assessment. SETTING: A rehabilitation unit in a medical school hospital. PATIENTS: Subjects consisted of 31 patients who suffered from stroke (right hemisphere lesions, 19; left hemisphere lesions, 12). MAIN OUTCOME MEASURES: Observation of behavior of hemi-inattention, ADL evaluation (transfer and gait), and conventional tests of USN (line bisection, line cancellation, copying of flowers) were conducted. USN test during stepping was performed by stepping on two force plates connected to a microprocessor and to videocassette player so that the subject was required to step continuously. RESULTS: Most of the subjects who had exhibited USN in the written tests on admission showed no defects in the test after about 60 days even if hemi-inattention was observed in behavior. In the test during stepping, subjects with hemi-inattention behavior neglected some visual stimulations applied from the direction contralateral to the hemisphere lesion. CONCLUSIONS: Combined tests of visual attention and distracter activity (stepping) can more effectively reflect the ADL functions of USN patients than conventional tests.     

Site-specific, photochemical proteolysis applied to ion channels in vivo

A method for site-specific, nitrobenzyl-induced photochemical proteolysis of diverse proteins expressed in living cells has been developed based on the chemistry of the unnatural amino acid (2-nitrophenyl)glycine (Npg). Using the in vivo nonsense codon suppression method for incorporating unnatural amino acids into proteins expressed in Xenopus oocytes, Npg has been incorporated into two ion channels: the Drosophila Shaker B K+ channel and the nicotinic acetylcholine receptor. Functional studies in vivo show that irradiation of proteins containing an Npg residue does lead to peptide backbone cleavage at the site of the novel residue. Using this method, evidence is obtained for an essential functional role of the "signature" Cys128-Cys142 disulfide loop of the nAChR alpha subunit.     

Echography at "high resolution" in the diagnosis of cervical lymphadenopathies in follow-up of thyroid carcinoma

BACKGROUND: In the diagnosis and follow-up of differentiated thyroid cancer (DTC) patients, a major clinical objective is to differentiate whether the cause of enlarged neck lymph nodes is either tumorous or inflammatory. High-frequency (7.5-10 MHz) ultrasound examination proved to be highly informative in this regard. An oval shape of the examined lymph node (longitudinal/anteroposterior diameter ratio, L/A > 1.5) and a central echogenic hilus indicate a benign lymphadenopathy. Roundness (L/A < or = 1.5), absence of the central echogenic hilus and a parenchymatous echogenicity are considered as signs of malignancy. METHODS: A series of 131 lymph nodes in DTC patients were examined at ultrasound and, on the basis of round shape (L/A < or = 1.5) in association with either one or both the other features (absence of central echogenic hilus, homogenous internal echo), 94 lymph nodes were classified a "malignant" and 37 "benign". The ultrasonographic diagnosis was evaluated with either cytologic or histologic diagnosis in all "malignant" nodes and in 7/30 "benign" nodes. In the other 30 "benign" nodes volume shrinkage or lymph node disappearance was observed during follow-up period. RESULTS: According to the results obtained, the ultrasound features utilized provided a good diagnostic accuracy (87%) with only 5 false positive case. CONCLUSIONS: High-frequency sonography, therefore, must be considered a useful and accurate method for evaluating suspected lymphadenopathy in DTC patients.     

The application of discriminant analysis for mapping cereals and pasture using object-based features

High mapping accuracies occur where crops differ spectrally (e.g.>90.0%; canola, corn, soybeans) and vice versa (e.g. <75.0%; cereals and pasture). Developing improved mapping methods has been an ongoing priority of Agriculture and Agri-Food Canada (AAFC) remote-sensing science. To this end, this study tests a data-driven object-based classification method using Discriminant Analysis (DA) method for mapping cereals and pasture from satellite data. In this approach, variables (number >400) derived from the image segmentation and object-based feature extraction of multi-date and multi-band optical (RapidEye) and microwave (RADARSAT-2) imagery were applied in a data-driven approach. We use in situ and satellite information collected over two study sites with different levels of heterogeneity (Winnipeg, Brandon) situated in the Canadian Prairies during the 2013 growing season to assess: (a) the type of DA model that most accurately classifies the cereals and pasture cover classes; and (b) how the classification accuracies obtained by the application of this DA model compare to those obtained from more traditional Maximum Likelihood (ML), Decision Tree (DT), and Random Forest (RF) classifications. We found that our DA-based approach was able to map cereals and pastures at our two study sites with the highest accuracies, but these accuracies did not improve significantly with the use of more complex DA model (including priori classification probabilities, more input principle components (PCs), the use of weights proportional to field area). Our results are encouraging for the wider application of the data-driven pre-processing of the inputs to the image classification by DA.