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A Gallagher

Publications and source records attributed to A Gallagher.

At least 19 recordsLinked to original sources

The expression of the EBV latent membrane protein (LMP-1) is independent of CD23 and bcl-2 in Reed-Sternberg cells in Hodgkin's disease.

A series of 33 cases of Hodgkin's disease was investigated for the presence of the EBV encoded latent gene product LMP-1 and of CD23 using immunohistochemical techniques. The expression of bcl-2 was examined in a subset of cases. LMP-1 was detected in the Reed-Sternberg cells in 15 cases. Although LMP-1 is known to upregulate CD23 and bcl-2, there was no correlation between the expression of LMP-1 and the detection of CD23 and bcl-2 in Reed-Sternberg cells.

Adolescent

Endothelin inhibits osteoclastic bone resorption by a direct effect on cell motility: implications for the vascular control of bone resorption.

The abundance of endothelin (ET)-producing endothelial cells in bone marrow and the proximity of these cells to bone-resorbing osteoclasts prompted us to evaluate the action of ET-1 on osteoclast function. Osteoclasts disaggregated from neonatal rat long bones were settled onto devitalized cortical bone substrate, and resorption was quantified by morphometry. The supernatant tartrate-resistant acid phosphatase activity was determined by a spectrophotometric method using paranitrophenol phosphate as substrate. Cell motility was quantified by time lapse video- and computer-assisted image processing using an empirical procedure for morphometric analysis. Cytosolic free calcium levels ([Ca2+]i) were measured in single cells by an indo 1-based microspectrofluorimetric method. Using the area of bone resorbed per slice as response, we found that ET-1 caused a significant (P = 0.011) concentration-dependent inhibition of osteoclastic bone resorption (EC50 = 2.5 nM) without inhibiting acid phosphatase secretion. Exposure of isolated osteoclasts to ET-1 also led to a marked concentration-dependent inhibition of osteoclast motility (EC50 = 7.9 nM; P = 0.013; t1/2 = 18 min) without significant effects on cell spread area. These effects of ET-1 were reversible after removing the peptide, and the cells remained viable during the experiments. In addition, ET-1 did not elevate [Ca2+]i at the concentrations tested. The results suggest that ET-1 specifically interacts with an osteoclast receptor to inhibit osteoclastic bone resorption and cell motility. As the concentration of ET-1 required for osteoclast inhibition was similar to that reported for smooth muscle contraction, it is possible that ET-1, produced locally from the bone marrow endothelial cell, might play a primary role in osteoclast regulation.

Acid Phosphatase

Criteria for the definition of Epstein-Barr virus association in Hodgkin's disease.

There is a clear association between the Epstein-Barr virus (EBV) and Hodgkin's disease (HD). EBV is not, however, detectable within the affected tissues of all cases. The proportion of positive cases varies from 15-79% depending on the assay used to detect EBV. The techniques utilised vary not only in sensitivity but in their ability to detect viral DNA, RNA, or protein and in their ability to demonstrate the cellular localisation of the virus. Thus, the biological significance of a positive result will vary depending on the method of analysis. In the present study, four different methods of detecting EBV were compared. RNA in situ hybridization was found to be the most practical method of detecting EBV in tumour cells. Using this assay EBV was detected in the Reed-Sternberg cells of 33% and 45% of the two series of HD cases examined in this study. We believe that these cases should be considered EBV-associated.

Adolescent

Viral involvement in Hodgkin's disease: detection of clonal type A Epstein-Barr virus genomes in tumour samples.

Thirty-five cases of Hodgkin's disease (HD) were analysed for the presence of Epstein-Barr virus (EBV) and human herpesvirus-6 (HHV-6) DNA. EBV genomes were detected in 11/35 cases while none of the cases was positive for HHV-6. Ten of the EBV-positive cases were subsequently analysed using a probe for the terminal region of the virus; the results suggested that the EBV-infected cells were clonally expanded. EBV subtypes specific DNA amplification was used to demonstrate that EBV subtype A, and not subtype B was present in the EBV-positive cases. The age distribution of the EBV-positive cases indicated a statistically significant trend for an increase in positivity with increasing age. This is the first indication that EBV is significantly associated with any subset of HD patients.

Adolescent

5-Aminosalicylic acid is a potent inhibitor of interleukin 1 beta production in organ culture of colonic biopsy specimens from patients with inflammatory bowel disease.

Interleukin 1 beta in biopsy specimens from inflamed colonic mucosa of patients with active inflammatory bowel disease was studied. Compared with normal colonic mucosal biopsy specimens, a significantly greater amount of interleukin 1 beta was present in rectal mucosa before (median (range) 4.3 (2.0-11.8) v 119.2 (30.1-286.8) pg/mg; p less than 0.01) and produced during organ culture (39.1 (9.4-106.8) v 97.6 (28.2-991.6) pg/mg; p less than 0.01). Values of interleukin 1 beta after culture correlated with concentrations of thromboxane B2. Organ culture of inflamed biopsy specimens in the presence of 5 aminosalicylic acid and dexamethasone reduced the amount of interleukin 1 beta detected. At the doses studied, 5 aminosalicylic acid also reduced the amount of leukotriene B4 detected after culture.

Adult

High circulating concentrations of interleukin-6 in active Crohn's disease but not ulcerative colitis.

Peripheral venous plasma concentrations of interleukin-6 were studied in 21 patients with active Crohn's disease, 20 patients with ulcerative colitis, and 16 control subjects. Interleukin-6 was detected in the plasma of 18 of 21 patients with Crohn's disease (median 47 (range less than 20-250) pg/ml) but in only two with ulcerative colitis and two control subjects. In the patients with Crohn's disease there was a significant negative correlation between the plasma interleukin-6 and the serum albumin concentrations. In eight patients with Crohn's disease and five patients with ulcerative colitis undergoing resection plasma from peripheral circulation and mesenteric vein draining diseased intestine was studied. Interleukin-6 was detected in seven of eight peripheral and mesenteric samples from the patients with Crohn's disease but was not detected in any of the samples from the patients with ulcerative colitis. There was no significant difference between mesenteric and peripheral samples in the concentrations of interleukin-6.

Adolescent

Detection of Epstein-Barr virus genomes in Hodgkin's disease: relation to age.

An investigation as to whether any particular subgroup of patients with Hodgkin's disease was particularly likely to be Epstein-Barr virus (EBV) genome positive was made on samples from 95 patients. These were grouped according to age and Hodgkin's disease subtype, and analysed using Southern blot analysis. Most samples from children or adults aged 50 years or over contained detectable EBV genomes; samples from young adults were only rarely positive. The differences in EBV positivity by age were highly significant, but there was no significant association between EBV and histological subtype after allowing for the effect of age. The results support the hypothesis that Hodgkin's disease in different age groups may have different aetiologies, and suggest that EBV does have a pathogenetic role in Hodgkin's disease in children and older age groups.

Adolescent

Plasma and tissue interleukin-2 receptor levels in inflammatory bowel disease.

Plasma and tissue interleukin-2 receptor (IL-2R) levels were determined in patients with active ulcerative colitis and Crohn's disease. Compared with healthy controls (median 440 U/ml; range 240-900), significantly higher levels of plasma IL-2R were present in patients with active ulcerative colitis (median 1180 U/ml; range 580-7150; P less than 0.002) and Crohn's disease (median 1340 U/ml; range 480-9000; P less than 0.002). Compared with other laboratory parameters, plasma IL-2R levels were related most closely to clinical score of disease activity in Crohn's disease. Plasma IL-2R levels also reflected the clinical course and may provide a more accurate assessment of disease activity in Crohn's disease. In plasma of patients undergoing intestinal resection of active inflammatory bowel disease, raised levels of IL-2R were present in samples from mesenteric vein (draining inflamed intestine) compared with those from peripheral vein. In tissue homogenates of colonic biopsies, significantly higher levels of IL-2R were present in specimens from colons with active ulcerative colitis compared with healthy controls (median 230.2, range 20.7-581.5 versus 77.9, range 34.2-291.3; P less than 0.02).

Biopsy

Suppress of the increase in free cytosolic calcium during the inhibition of T-cell activation by an autoantibody present in the serum of leprosy patients.

A serum factor, believed to be an IgG autoantibody, in certain patients with lepromatous leprosy inhibits the proliferation of mitogen-stimulated lymphocytes. To investigate which stage of the cell cycle was inhibited, we examined the effect of these sera on the kinetics of lymphocyte activation induced by several mitogenic agents: phytohaemagglutinin (PHA), the calcium ionophore A23187, the phorbol ester phorbol myristate acetate (PMA) and purified protein derivative of BCG (PPD). Seven out of 54 sera tested were found to inhibit PHA-stimulated proliferation. Inhibitory sera and to a lesser extent serum IgG from leprosy patients were capable of suppressing the increase in free cytosolic calcium normally observed immediately after PHA stimulation. Subsequent stages of the cell cycle, increase in cell size, the expression of the IL-2 receptor and increase in DNA were also suppressed. The inhibitory sera was not toxic and, if addition of the sera was delayed, would not inhibit lymphocytes that had already entered the cell cycle. Using mitogenic agents which act intracellularly, the normal early increase in cell size with A23187- and PMA-stimulated lymphocytes was not affected by inhibitory leprosy sera or serum IgG, but all subsequent steps in the cell cycle were suppressed; although the inhibition of proliferation in PMA-stimulated cultures was incomplete. The mechanism of action of the inhibitory sera and derived IgG, although acting through a cell surface antigen, appears to interfere with a fundamental process in activation since the effect was seen with all of the diverse stimuli examined in this study.

Autoantibodies

Care--old style.

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Child

Comparison of the chemotactic behaviour of Rhizobium leguminosarum with and without the nodulation plasmid.

The chemotactic behaviour of a strain of Rhizobium leguminosarum biovar viciae was investigated. The flavanoids apigenin and naringenin, inducers of transcription of the nodulation (nod) genes, were both potent attractants but hesperitin, another flavone nod gene inducer, was not. The response of strains containing the Sym plasmid pRL1Jl to apigenin and naringenin was significantly greater than the response of a strain cured of the plasmid, although both strains gave a positive response. Addition of the flavanol kaempferol, an antagonist of nod gene induction, had no detectable effect on the chemotactic response to naringenin or apigenin, but was itself found to be an attractant. The attractant response to a variety of amino acids and sugars was not affected by the presence of the Sym plasmid. Homoserine, the most abundant nitrogenous compound in legume exudates, was also found to be an attractant. However, although the Sym plasmid is required for the biovar to metabolize homoserine as a carbon source, it was not required for the chemotactic response. A group of membrane proteins showed increased methylation in response to stimulation with serine. There was no measurable change in methylation after stimulation with apigenin.

Bacterial Proteins

Changes in activation markers and cell membrane receptors on human peripheral blood T lymphocytes during cell cycle progression after PHA stimulation.

Phytohaemagglutinin (PHA)-stimulated peripheral blood lymphocytes were examined sequentially for changes in volume, the appearance of cell membrane receptors and nucleic acid synthesis. The kinetics of appearance of activation antigens were compared with the progress of the cell through the separate events of volume growth and nucleic acid syntheses, to determine points at which regulation of receptors may control further progress through the cell cycle. In all samples tested there was a consistent pattern of response in the proportion of cells progressing through the cell cycle. Most of the T cells increased in size (mean 82% at 24 hr), fewer cells entered the Gla/Glb phase with the onset of RNA synthesis (mean 68% at 48 hr) and even fewer entered DNA synthesis (mean 42% at 72 hr). The time-course of appearance and the number of cells expressing IL-2 receptors were almost identical with that of cells responding by RNA synthesis. A similar correlation was observed between expression of the transferrin receptor and DNA synthesis. Addition of anti-Tac antibody temporarily suppressed the onset of RNA synthesis and antibodies to the transferrin receptor suppressed DNA synthesis. These linkages are further evidence that IL-2 and transferrin are the specific signals for cellular RNA and DNA synthesis. With optimal concentrations of PHA, addition of IL-2 did not increase the proportion of cells bearing activation antigens or undergoing nucleic acid synthesis. Suboptimal concentrations of PHA produced a small reduction in the number of cells expressing the IL-2 receptor, but a much greater reduction in the rate of entry into RNA synthesis. There was a consistent increase in all activation parameters tested with the addition of IL-2, but the proportion of cells expressing the transferrin receptor and entering DNA synthesis was consistently lower than that of cells that expressed the IL-2 receptor or entered RNA synthesis. This suggests that regulation of the IL-2 receptor is not responsible for the reduction in the number of cells that proceed to proliferation. The CD2 antigen (T11(1] showed increasing expression in a step-wise fashion after activation, the increases coinciding with the onset of RNA and DNA syntheses.

Cell Cycle

Dietary patterns and metabolic control in diabetic diets: a prospective study of 51 outpatient men on unmeasured and exchange diets.

In a prospective, single blind study, 51 adult diabetic, male outpatients at or below ideal body weight (IBW), all but four of whom were insulin-treated, were randomly assigned to a calorically defined exchange diet (EXCH) or an unmeasured diet avoiding refined sugars (UNMEAS). Fasting chemistries, weights and 48-hr dietary recalls were obtained every 3 months for 3 years. There was no difference between groups in mean body weight, mean caloric intake, percentages of carbohydrate or fat intake, day to day consistency of caloric intake, or fasting chemistries. The patients whose actual weight tended to be less than IBW consumed more calories than their theoretical calculated needs (p = 0.002) without effect on glycemic control or stability of weight. Patients with low carbohydrate intake (less than 30%) had significantly higher triglyceride levels (p = 0.015). Otherwise, variations in dietary patterns were not reflected in fasting clinical chemistries. Individual fasting serum glucose levels were not related to consistency of carbohydrate intake, caloric distribution throughout the day or composition of diets. These results suggest that actual dietary patterns of subjects following exchange diabetic diets with precise caloric prescriptions are similar to those resulting from unmeasured diets with restriction of simple sugars. Other than correlation between carbohydrate intake and triglycerides, there was no detectable effect of variations in dietary patterns upon conventional chemical metabolic levels.

Adult

The yeast mitochondrial ATPase complex. Subunit composition and evidence for a latent protease contaminant.

1. The subunit compositions of the F1 (oligomycin-insensitive) and F1--F0 (oligomycin-sensitive) mitochondrial ATPase complexes from Saccharomyces cerevisiae have been examined by the highly resolving technique of sodium dodecyl sulphate-polyacrylamide slab gel electrophoresis using a discontinuous buffer system. When isolated in the presence of protease inhibitors, F1 and F1--F0 contained five and twelve bands, respectively; this contrasts with the four- and ten-band patterns seen previously using the less resolving disc gel method. When isolated in the absence of protease inhibitors both F1 and F1--F0 contain spurious polypeptides produced by proteolytic modification. 2. Endogenous protein turnover in S. cerevisiae was impaired in the presence of protease inhibitors. F1--F0 isolated from cells grown in the presence and absence of inhibitors contained an identical polypeptide composition, suggesting that the subunits are not significantly modified by endogenous proteases prior to cell harvesting. 3. Yeast F1--F0 prepared in the presence of protease inhibitors contains a latent, sodium dodecyl sulphate-activated protease contaminant. Sodium dodecyl sulphate-induced proteolysis is largely confined to the 52 000 dalton alpha subunit which degrades into polypeptides of 40 000 and 10 700 daltons. The 40 000 dalton band is apparently equivalent to the polypeptide previously designated subunit 3. 4. Both F1 and F1--F0 were isolated from Torulopsis glabrata, a yeast with considerably shorter mitochondrial DNA than that in S. cerevisiae. F1--F0 catalysed high rates of ATP--32Pi exchange when reconstituted into phospholipid vesicles, thus demonstrating the presence of a complete coupling mechanism. F1--F0 contained approximately twelve subunits and F1 five, like the S. cerevisiae complexes. It therefore appears that the shorter mitochondrial DNA length does not produce a significantly simpler ATPase subunit structure.

Adenosine Triphosphatases

Reducing diets in rats.

Adult obese rats were restricted to equal intakes provided by four diets which varied widely in protein, fat and carbohydrate content. Loss in body weight and body fat was similar in all groups. The only significant dietary effect observed was a modest increase in the body protein content of the animals fed the high protein diets. Low carbohydrate--high fat--high protein diets, as expected, showed no advantage as reducing diets when compared to other diets on an equal calorie basis.

Animal Nutritional Physiological Phenomena