P-selectin mediates adhesion of the human melanoma cell line NKI-4: identification of glycoprotein ligands

Activated endothelial cells and stimulated platelets express the cell adhesion molecule P-selectin (CD62P), which mediates adhesion to various leukocytes and certain types of cancer cells. In this study, we show Ca2+-dependent binding of P-selectin to NKI-4 cells, a cell line derived from a human melanoma. The binding is inhibited by P7 (a leukocyte adhesion blocking mAb against P-selectin), but not by PL5 (a leukocyte adhesion blocking mAb against P-selectin glycoprotein ligand-1; PSGL-1). Further, expression of PSGL-1 could not be detected on NKI-4 cells by either PL5 mAb or an Ab against a synthetic peptide corresponding to a portion of the PSGL-1 sequence. P-selectin affinity chromatography of lysates from in vivo [3H]-glucosamine-labeled NKI-4 cells resulted in the isolation of three glycoproteins, with apparent molecular masses of approximately 250, approximately 110, and approximately 100 kDa under reducing conditions and approximately 230, approximately 105, and approximately 85 kDa under nonreducing conditions. These molecules could be precipitated by P-selectin, but not by E-selectin. EDTA and the P7 mAb, but not the PL5 mAb, inhibited the binding of P-selectin to the purified ligands. Surprisingly, we found that sodium chlorate, a sulfation inhibitor, did not inhibit the binding of P-selectin to NKI-4 cells and that [35S]-sulfate did not label the NKI-4 cell ligands. We conclude that P-selectin-dependent adhesion of the human melanoma cell line NKI-4 is mediated by a novel class of glycoprotein ligands.     

Measurement of histamine release from human lung tissue ex vivo by microdialysis technique

OBJECTIVE AND DESIGN: Currently no method is available for measurement of mediator release from intact human lung. In this study, a microdialysis technique was used to measure histamine release from mast cells in human lung tissue ex vivo. MATERIAL: Microdialysis fibers of 216 microm were inserted into lung tissue and perfused with Krebs Ringer buffer at a rate of 3 microl/min. After a 15 min period of steady-state perfusion, anti-IgE and vehicle were injected into the lung tissue above individual fibers. Samples from each fibre were collected for 20 min at 2 min intervals. Histamine was assayed fluorometrically. RESULTS: Anti-IgE concentrations of 40-40,000 U/ml dose-dependently released histamine, significant histamine release being demonstrated with anti-IgE concentrations of 400 U/ml and greater. The kinetics of histamine release showed peak values 2-8 min after the injection. Great individual responses were observed but data could be reproduced within individual donors. Monocyte chemoattractant protein-1, a potent basophil secretagogue, did not induce histamine release in lung tissue which indicated mast cells to be the histamine source. Substance P did not release histamine in the lung tissue. CONCLUSIONS: The microdialysis technique allowed measurements of histamine release from mast cells in intact lung ex vivo. The method may prove useful since a number of experiments can be performed in a few hours in intact lung tissue without any dispersion or enzymatic treatment.     

Neuroleptic use and behavioral disturbance in nursing homes: a 1-year study

This article discusses a longitudinal study of change in disruptive behaviors among nursing home residents treated with neuroleptics compared with those not treated with neuroleptics. Observations were made of 201 participants on admission to and after 1 year in eight skilled nursing facilities. Nine disruptive behaviors were measured using the Psychogeriatric Dependency Rating Scale with nursing assistants. Neuroleptic use was documented from medication records. Odds ratios are reported for the association of behavior at baseline and use of neuroleptics on nine problem behaviors. For those who received neuroleptics during the year, there was greater change in both developing and resolving disruptive behaviors than for those not receiving neuroleptics. For both groups, restless or pacing behavior and belligerent behavior manifested by refusing instructions changed the most, both in developing and in apparently resolving. Our results show that change in disruptive behaviors occurs among nursing home residents regardless of neuroleptic use, but it occurs more frequently among those who receive neuroleptic medication. Knowledge of which disruptive behaviors are most likely to resolve or develop is important in training nursing home staff to cope with the behaviors as well as in planning interventions that may modify such behaviors.     

Crystal structure of GCN4-pIQI, a trimeric coiled coil with buried polar residues

BACKGROUND AND PURPOSE: Early and accurate diagnosis of brain edema in stroke patients is essential for the selection of appropriate treatment. We examined the correlations between the changes in the apparent diffusion coefficient (ADC), regional water content, and tissue ultrastructure during early focal cerebral ischemia. METHODS: The left middle cerebral arteries of cats were occluded with an intramagnet occlusion/recirculation device. T2-weighted, diffusion-weighted, and perfusion imaging were performed repeatedly during the initial 3 hours after occlusion. The ADCs obtained from ADC maps were compared with the corresponding tissue water content values determined by gravimetry and electron microscopic water localization. RESULTS: ADC reduction was detected in areas of low perfusion 15 minutes after occlusion and thereafter. The water content increase correlated linearly with the ADC decreases in both the gray and white matter. However, both the water content corresponding to an ADC value and the rate of ADC change of the gray and white matter differed significantly (P<.05) as follows: y = -10105x + 8533 (r=.86) and y = -6174x + 4611 (r=.67), respectively, where x is the water content (grams water per gram tissue) and y is the ADC (x 10(-6) mm2/s). Hydropic astrocytic swelling was seen in both structures, and in the white matter, oligodendroglial and myelinated axonal swelling and periaxonal space enlargement were observed. CONCLUSIONS: When early ischemic edema in experimental focal cerebral ischemia is evaluated with ADC mapping, the different slopes and intercepts of the water content and ADC correlation lines for the gray and white matter, which probably reflect different ultrastructural localization of water, should be taken into account.     

Cardiac dysrrhthmia in systemic disease

This is an ECG of a 6-year-old girl. She presented to us for management of epilepsy. Magnetic resonance imaging (MRI) of the brain showed multiple lesions consistent with cortical and subcortical tubers. There were also achromic spots on her skin and echocardiography demonstrated a rhabdomyoma near the right ventricular outflow tract. A diagnosis of tuberous sclerosis, an autosomal-dominantly inherited condition, was made. What is abnormal about the ECG?     

Plumbagin,a Biomolecule with (Anti)Osteoclastic Properties

Plumbagin is a plant-derived naphthoquinone that is widely used in traditional Asian medicine due to its anti-inflammatory and anti-microbial properties. Additionally, plumbagin is cytotoxic for cancer cells due to its ability to trigger reactive oxygen species (ROS) formation and subsequent apoptosis. Since it was reported that plumbagin may inhibit the differentiation of bone resorbing osteoclasts in cancer-related models, we wanted to elucidate whether plumbagin interferes with cytokine-induced osteoclastogenesis. Using C57BL/6 mice, we unexpectedly found that plumbagin treatment enhanced osteoclast formation and that this effect was most pronounced when cells were pre-treated for 24 h with plumbagin before subsequent M-CSF/RANKL stimulation. Plumbagin caused a fast induction of NFATc1 signalling and mTOR-dependent activation of p70S6 kinase which resulted in the initiation of protein translation. In line with this finding, we observed an increase in RANK surface expression after Plumbagin stimulation that enhanced the responsiveness for subsequent RANKL treatment. However, in Balb/c mice and Balb/c-derived RAW264.7 macrophages, these findings could not be corroborated and osteoclastogenesis was inhibited. Our results suggest that the effects of plumbagin depend on the model system used and can therefore either trigger or inhibit osteoclast formation.     

Morphological and microchemical phenomena in the high-temperature oxidation of binary Al-Li alloys

The high-temperature oxidation behavior of binary Al-Li alloys has been characterized by scanning electron microscopy and secondary ion mass spectrometry in order to understand the mechanism of rapid oxidation in these alloys and to correlate the oxide morphology to its microchemistry. The oxide scale developed on polished specimens during short exposures in air at 530°C shows characteristic nodules that usually nucleate at grain boundaries. Examination of the alloy surface after removal of the oxide layer shows that the initial growth of the oxide nodules occurs laterally in addition to thickening normal to the oxide/alloy interface. Microchemical analysis of the oxide film with a scanning ion microprobe reveals a thick Li-oxide layer at the oxide/gas interface indicating preferential oxidation of Li at the free surface; the rest of the oxide film is composed of both Al- and Li-rich oxides, probably Li₂O and LiAlO₂ The presence of trace impurities (K, Na, F, and Cl) in the oxide scale was also detected. A microstructural model for the development of the oxide film in the Al-Li system is presented on the basis of both morphological and microanalytical data obtained in this study; this new model is compared with existing models.     

Text-dependent speaker verification using classical LBG,adaptive LBG and FCM vector quantization

An important task of speaker verification is to generate speaker specific models and match an input speaker’s utterance with these models. This paper focuses on comparing the performance of text dependent speaker verification system using Mel Frequency Cepstral Coefficients feature and different Vector Quantization (VQ) based speaker modelling techniques to generate the speaker specific models. Speaker-specific information is mainly represented by spectral features and using these features we have developed the model which serves as an important entity for determining the claimed identity of the speaker. In the modelling part, we used Linde, Buzo, Gray (LBG) VQ, proposed adaptive LBG VQ and Fuzzy C Means (FCM) VQ for generating speaker specific model. The experimental results that are performed on microphonic database shows that accuracy significantly depends on the size of the codebook in all VQ techniques, and on FCM VQ accuracy also depend on the value of learning parameter of the objective function. Experiment results shows that how the accuracy of speaker verification system is depend on different representations of the codebook, different size of codebook in VQ modelling techniques and learning parameter in FCM VQ.     

Experimental investigation of impact of diesel particulate filter on smoke and NO<Subscript>x</Subscript> emissions of a Euro-I compression ignition engine with active and off-board regeneration

Old engines (Euro III or earlier) produce more emissions, and it will be difficult to entirely stop their usage especially in developing and under-developed nations; hence, it is desired that appropriate emission reduction technologies are tested on such engines to analyze their feasibility and economical acceptability. While most such studies have been conducted on constant speed stationary engines and modern engines, this study tried to analyze the effectiveness of an uncoated wall-flow type ceramic diesel particulate filter on a Euro-I, water-cooled, direct injection, variable speed, compression ignition engine in a laboratory set-up in India. Also, this study focused on diesel particulate filter regeneration by two methods: active regeneration by diesel injection in the particulate filter using an electronic control unit and off-board regeneration by taking out and heating the diesel particulate filter in an electrical resistance furnace at 650 °C for 10 h. The results, in the form of smoke emission, NO_x emission and engine performance, obtained using both the regeneration methods were analyzed, and conclusions were drawn. It was found that using diesel particulate filter, particulate matter emissions (smoke) were almost entirely eliminated. It was also found that off-board regeneration had numerous advantages compared to active regeneration. Since a furnace would be needed for off-board regeneration, an exchange process for diesel particulate filter is suggested.