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J R Baker

Publications and source records attributed to J R Baker.

At least 73 records · Page 4Linked to original sources
PubMed1999

Characterization of FAP-1 expression and function in thyroid follicular cells.

Human thyrocytes are resistant to Fas-mediated programmed cell death (PCD). It has been reported that a labile protein inhibitor is involved in the protection of thyrocytes from PCD, and its action can be reversed by incubation of thyrocytes with cycloheximide (CHX) during treatment with agonist anti-Fas Ab. Fas-associated phosphatase-1 (FAP-1) is a protein that has been shown to interact with the negative regulatory domain of Fas and block Fas-mediated apoptosis in FAP-1 transfected Jurkat cells. We investigated the possibility that FAP-1 might be involved in protection against Fas-mediated PCD in human thyrocytes. FAP-1 mRNA was detected in primary thyrocytes using a ribonuclease protection assay. The presence of FAP-1 protein was confirmed by immunohistochemical staining and flow cytometry using a polyclonal anti-FAP-1 Ab. FAP-1 protein also disappeared from thyroid cells in response to CHX. To determine whether FAP-1 is a functional inhibitor of PCD in thyrocytes, we incubated thyrocytes with synthetic SLV (Ac-SLV) tripeptide to compete with Fas for interaction with FAP-1. Thyrocytes treated with Ac-SLV tripeptide showed significantly increased cell death as compared to cells treated with control tripeptide. In addition, in the presence of a suboptimal concentration of CHX, the Ac-SLV tripeptide yielded a strong, synergistic increase in Fas-mediated PCD as compared to thyrocytes treated with control tripeptide. These results implicate FAP-1 as a regulator of Fas-induced PCD in thyrocytes.

Apoptosis↗
PubMed1999

1Alpha,25-dihydroxyvitamin D3 up-regulates Bcl-2 expression and protects normal human thyrocytes from programmed cell death.

Apoptosis is thought to play an important role in the pathogenesis of autoimmune thyroid disease. 1alpha,25-dihydroxyvitamin D3 (VD3) has been shown to suppress several autoimmune diseases. However, the mechanism by which VD3 has these effects is not known. We evaluated the alterations in apoptosis, induced by VD3. Thyrocytes were treated with VD3, and the expression of the Bcl-2 family molecules was studied at both the messenger RNA and protein levels. It was found that VD3 significantly induced the expression of Bcl-2 messenger RNA and protein in thyrocytes but had no effect on the expression of Bcl-xl and Bax. The increase in Bcl-2 expression, mediated by VD3, correlated with protection of thyrocytes against the induction of apoptosis by either staurosporine or UV irradiation. VD3-induced increases in the expression of Bcl-2 could be mimicked by VD3 analogs with high nuclear receptor affinity, but not by analogs only with nongenomic actions. These data indicate a role for Bcl-2 in the regulation of apoptosis in thyrocytes and raise the possibility that VD3 or its agonists may have therapeutic benefit in thyroid disorders.

Apoptosis↗
PubMed1999

Fas (CD95) expression is up-regulated on papillary thyroid carcinoma.

Thyrocyte apoptosis signaled through the Fas receptor has been proposed as a mechanism for the cytotoxicity observed in thyroiditis, but the role the Fas pathway plays in thyroid cancer is not known. We examined Fas expression in thyroid tissue derived from patients with papillary carcinoma and follicular cancer. More intense immunohistological staining for the Fas protein was observed on papillary cancer cells as compared with adjacent normal follicles. To further characterize the expression of Fas in papillary cancer, paired normal and cancerous thyroid tissues were obtained at thyroidectomy from several donors, digested, and placed into cell culture. Messenger RNA was analyzed by ribonuclease protection assays, and protein was identified by flow cytometry. Fas expression was detected at levels up to 3-fold higher in cancerous thyrocytes compared with paired normal cells. To determine whether the expressed Fas antigen was functional, thyrocytes were treated with a monoclonal IgM anti-Fas antibody (clone CH11; Upstate Biotechnology, Inc., Lake Placid, NY) in the presence of interferon-gamma and cycloheximide. Whereas both normal and cancerous thyrocytes were induced to die after this treatment, the cancerous thyrocytes were more sensitive to anti-Fas antibody. This work demonstrates that the Fas antigen is expressed and functional on papillary thyroid cancer cells and this may have potential therapeutic significance.

Adenocarcinoma, Follicular↗
PubMed1999

The role of Fas-mediated apoptosis in thyroid autoimmune disease.

Apoptosis is a carefully regulated mechanism of cell death that differs from necrosis and plays an important role in normal tissue development and homeostasis, as well as disease processes. Apoptosis also plays an important role in autoimmunity. Defective apoptosis can cause systemic autoimmunity by allowing the survival of autoreactive lymphocytes. It may also be involved in the pathogenesis of organ-specific autoimmune diseases, such as Hashimoto's thyroiditis, through altered target organ susceptibility. Apoptosis signaling pathways can be initiated through activation of death receptors. One of these pathways employs the death receptor Fas and its ligand (FasL). Fas expression and death pathway signaling have been demonstrated in the thyroid, but there is controversy surrounding the expression of FasL and its role in thyroid autoimmunity. A number of proteins, including FAP-1, Bcl-2 and I-FLICE may regulate the Fas pathway in the thyroid and provide potential mechanisms for modifying the pathogenesis of autoimmune thyroid disease.

Animals↗
PubMed1998

Homomultimeric protease in the hyperthermophilic bacterium Thermotoga maritima has structural and amino acid sequence homology to bacteriocins in mesophilic bacteria.

A novel homomultimeric protease (> 669 kDa), based on 31 kDa subunits, was purified from cell extracts of the hyperthermophilic bacterium Thermotoga maritima. This protease exhibits activity toward chymotrypsin and trypsin substrates, optimally at 90 degrees C and pH 7.1, and has a half-life of 36 min at 95 degrees C. Transmission electron microscopy established that the protease consists of a large globular assembly which appears circular from the front view. The function of this protease in T. maritima remains unclear, although putative homologs include a 29 kDa antigen from Mycobacterium tuberculosis and a 31 kDa monomer of a high molecular weight bacteriocin produced by Brevibacterium linens [Valdes-Stauber, N. and Scherer, S. (1996) Appl. Environ. Microbiol. 62, 1283-1286]. The relationship of these mesophilic proteins to the T. maritima protease suggests that their antibacterial activity may involve elements of proteolysis, and raises the prospect for antimicrobial ecological strategies in hyperthermophilic niches.

Amino Acid Sequence↗
PubMed1998

Causes of mortality and non-fatal conditions among grey seals (Halichoerus grypus) found dead on the coasts of England, Wales and the Isle of Man.

A survey of the diseases detectable in 141 grey seals stranded on the coasts of England and Wales away from breeding colonies was carried out between mid-1989 and early 1997. The most common fatal conditions in pups less than three weeks of age were trauma (24 per cent of deaths) and dystocia (12 per cent); in pups more than three weeks of age thy were starvation (22 per cent) and pneumonia (22 per cent); in juveniles they were drowning in fishing gear (30 per cent) and starvation (19 per cent), and in adults a variety of respiratory diseases were the most common causes of death (45 per cent). Many other diseases, both fatal and non-fatal, were recorded.

Animals↗
PubMed1998

Efficient transfer of genes into murine cardiac grafts by Starburst polyamidoamine dendrimers.

Starburst dendrimer, a structurally defined, spherical macromolecule composed of repeating polyamidoamino subunits, was investigated to augment plasmid-mediated gene transfer efficiency in a murine cardiac transplantation model. The grafts were directly injected with naked pCH110, a plasmid encoding beta-galactosidase (beta-Gal), or pCH110-dendrimer complex, and reporter gene expression determined by X-Gal staining. The grafts injected with pCH110-dendrimer demonstrated widespread and extended beta-Gal expression in both myocytes and the graft infiltrating cells from 7 to 28 days, compared to the grafts injected with naked pCH110 that expressed beta-Gal only in myocytes for less than 14 days. p alphaMHC-vIL-10, as plasmid encoding viral interleukin-10 (vIL-10) under the control of alpha-myosin heavy chain promoter, was able to prolong allograft survival from 13.9 +/- 0.9 days to 21.4 +/- 2.3 days (p < 0.005). When dendrimer G5EDA was used with p alphaMHC-vIL-10, 60-fold less DNA resulted in significant prolongation of graft survival to 38.6 +/- 4.7 days (p < 0.0005). The dose of DNA, the charge ratio of DNA to dendrimer, and the size generation of the dendrimers were all determined to be critical variables for prolongation of allograft survival in this model system. Thus, the use of the Starburst dendrimer dramatically increased the efficiency of plasmid-mediated gene transfer and expression. Production of immunosuppressive cytokines at higher amounts for longer periods of time in a greater expanse of tissue enhanced the immunosuppressive effect and prolonged graft survival further.

Animals↗
PubMed1998

Imaging subcortical auditory activity in humans.

There is a lack of physiological data pertaining to how listening humans process auditory information. Functional magnetic resonance imaging (fMRI) has provided some data for the auditory cortex in awake humans, but there is still a paucity of comparable data for subcortical auditory areas where the early stages of processing take place, as amply demonstrated by single-unit studies in animals. It is unclear why fMRI has been unsuccessful in imaging auditory brain-stem activity, but one problem may be cardiac-related, pulsatile brain-stem motion. To examine this, a method eliminating such motion (using cardiac gating) was applied to map sound-related activity in the auditory cortices and inferior colliculi in the brain stem. Activation in both the colliculi and cortex became more discernible when gating was used. In contrast with the cortex, the improvement in the colliculi resulted from a reduction in signal variability, rather than from an increase in percent signal change. This reduction is consistent with the hypothesis that motion or pulsatile flow is a major factor in brain-stem imaging. The way now seems clear to studying activity throughout the human auditory pathway in listening humans.

Auditory Cortex↗
PubMed1998

A localized double-quantum filter for the in vivo detection of brain glucose.

A double-quantum filter (DQF) sequence with PRESS localization was developed for in vivo detection of the glucose resonances in the 3.85-ppm region of the brain proton spectrum. The efficiency and spectral editing characteristics were studied in phantom and animal experiments. Approximately 45% detection efficiency was achieved at 4.7 T with TE = 68 ms. Since the efficiency of the DQF method is dependent on the relative phases of the RF pulses, a phase calibration procedure was used to correct for phase shifts induced by the spatial localization. In addition to detecting the 3.85-ppm glucose resonances with approximately 45% efficiency, the DQF sequence simultaneously detects 1.3-ppm lactate resonance with approximately 20% efficiency. The use of the DQF technique for simultaneously monitoring both the input and output of anaerobic glycolysis in the brain was demonstrated by detecting brain glucose and lactate in the same acquisition after iv injection of glucose followed by the induction of global ischemia.

Animals↗
PubMed1998

Apoptosis and thyroiditis.

The origin of the various forms of autoimmune thyroiditis remains unclear. Most investigations into the pathogenesis of these disorders have focused on immune abnormalities that might lead to an autoimmune response. However, no unique immune response to thyroid autoantigens has been identified that either is limited to patients with thyroiditis or is absolutely correlated with clinical disease expression. CD8 T-cell-mediated cytotoxicity is thought to be a major cause of thyroid follicular cell damage in thyroiditis. This damage is produced in part through the induction of apoptosis in thyroid cells. Recent studies have demonstrated that programmed cell death is regulated in thyroid cells and that a major pathway for immune-mediated apoptosis, the Fas pathway, is blocked by labile inhibitors in a manner that could prevent cytotoxicity. This review also examines several other types of regulation of apoptotic pathways in thyrocytes. We hypothesize that the regulation of programmed cell death pathways in the thyroid may alter the expression of autoimmune thyroid diseases by modifying the susceptibility of thyroid cells to immune-mediated apoptosis.

Apoptosis↗
PubMed1998

The effect of synthetic surfactant Exosurf on gene transfer in mouse lung in vivo.

Gene transfer in the lung holds promise for the treatment of diseases such as pulmonary fibrosis, cystic fibrosis and asthma. Pulmonary surfactant has been reported to enhance expression from endobronchial, adenovirus-mediated gene transfer in experimental animals. This study examines the effect of exogenous synthetic surfactant (Exosurf) on gene expression from naked plasmid DNA administered endobronchially to adult mice. Transfection efficiency was evaluated by quantifying the expression of chloramphenicol acetyltransferase (CAT) and luciferase (Luc) genes in the lung. Endobronchial administration of either CAT or Luc expression plasmid DNA resulted in detectable concentrations of each reporter protein. CAT expression from plasmid DNA was monitored after endobronchial administration with the maximal expression observed at 3-5 days after administration and decreasing for 5 days thereafter. When DNA was delivered in a 50% suspension of Exosurf, the expression of either CAT or Luc was significantly reduced by 89.6 +/- 1.4% and 82.7 +/- 10.5%, respectively. The decrease in Luc expression was closely correlated (r = 0.99, P < 0.001) to log concentration of surfactant in the plasmid buffer solution (IC50 = 8.6%). CAT expression was not altered when surfactant was administered either 2 h before or after plasmid DNA instillation. Examination of the components of Exosurf revealed that two compounds, DPPC and tyloxapol, showed inhibitory effects on CAT expression. However, the inhibition caused by Exosurf appeared greater than that of either component. Our results suggest that the lung surfactant is a barrier to transfection of the endobronchial airway and may be partly responsible for the low expression of exogenous DNA in vivo in the bronchial tree.

Administration, Topical↗
PubMed1998

Accuracy of devices used for self-monitoring of blood glucose.

We measured the inaccuracy of 17 home blood glucose monitors (two visually read, eight colorimetric and seven amperometric). Using strips from a single batch, blood glucose measurements were performed by three medical laboratory technologists on at least 50 capillary blood specimens from patients attending two diabetes clinics. Additional capillary blood was deproteinized and assayed with hexokinase to give a whole blood glucose result. A dedicated glucose analyser was also studied to cross-validate the methodology. At a mean glucose concentration of about 9 mmol/L, monitor readings differed from the reference results by -5.1 to +19.5% with three systems failing to meet the American Diabetes Association guideline for total error of less than 15%. This problem would be alleviated by manufacturers adopting a common policy on calibration and on reporting as a plasma or whole blood value.

Ambulatory Care↗
PubMed1997

Autoimmune endocrine disease.

Autoimmune endocrine diseases are serious disorders that utilize immense health care resources and cause tremendous disability. They include type 1 diabetes mellitus, thyroiditis, Graves disease, Addison disease, and polyglandular syndromes. Analysis of the basis of autoimmune diseases has been aided by the application of new knowledge in immunologic physiology. Recent investigations using these techniques have revealed complicated disorders that have varied pathogenesis and complex genetic predispositions. While the mainstay of treatment for these diverse diseases remains the replacement of hormones produced by the damaged endocrine organ, investigations into the pathogenesis of these disorders provide hope for the development of specific therapeutic measures to block their pathologic basis.

Addison Disease↗
PubMed1997

Antibodies to Brucella in marine mammals around the coast of England and Wales.

Following the isolation of previously unrecognised species of Brucella from stranded seals and cetaceans in Scotland and northern England, a serological survey was carried out to investigate the range of marine mammal species which may have been exposed to Brucella species around the coasts of England and Wales, the prevalence of infection and the temporal and geographical distribution of seropositive animals. Serum collected from 153 stranded marine mammals from the coasts of England and Wales between 1989 and 1995 were tested by competitive and indirect ELISA. Positive titres were recorded for six of 62 (10 per cent) grey seals (Halichoerus grypus), one of 12 (8 per cent) common seals (Phoca vitulina), 11 of 35 (31 per cent) harbour porpoises (Phocoena phocoena) and nine of 29 (31 per cent) common dolphins (Delphinus delphis) tested. Positive titres were also found in a striped dolphin (Stenella coeruleoalba), a bottlenose dolphin (Tursiops truncatus), a killer whale (Orcinus orca) and a pilot whale (Globicephala melas). The seropositive animals were from all around the coasts of England and Wales and the first seropositive sample was from a common dolphin in 1990.

Animals↗
PubMed1997

Specificity of the hyaluronate lyase of group-B streptococcus toward unsulphated regions of chondroitin sulphate.

The purification and properties of a hyaluronate lyase secreted by Streptococcus agalactiae, which is believed to facilitate the invasion of host tissues by the organism, have been described previously [Pritchard, Lin, Willingham and Baker (1994) Arch. Biochem. Biophys. 315, 431-436]. The specificity of the limited cleavage of chondroitin sulphate by the enzyme is the subject of this report. To simplify the task, a chondroitin sulphate from the Swarm rat chondrosarcoma, which contains only 4-sulphated and unsulphated disaccharide repeats, was used in this study. Tetrasaccharides from an ovine testicular hyaluronidase digest of the chondroitin sulphate were isolated, identified and tested as substrates of the streptococcal hyaluronate lyase. Only tetrasaccharides with an unsulphated disaccharide at the reducing end were cleaved (by elimination at the N-acetylgalactosaminidic bond). Thus chondroitin sulphate chains are cleaved by the action of this lyase at every unsulphated disaccharide repeat, but release of unsaturated unsulphated disaccharides only occurs from sites where two or more sequential unsulphated disaccharide repeats are present. Analysis of the chondrosarcoma chondroitin sulphate showed that of approximately five unsulphated disaccharide repeats per chain, two are clustered. The ability of group-B streptococcal hyaluronate lyase to cleave chondroitin sulphate may allow the organisms to invade tissues more efficiently. The demonstrated specific and highly limited cleavage of chondroitin sulphate by this bacterial lyase promises to be a useful tool in the determination of chondroitin sulphate structure and variability.

Animals↗