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Biomedical subjects

T Cole

Publications and source records attributed to T Cole.

At least 73 records · Page 4Linked to original sources

Reduction of disulfide bonds in an amyloidogenic Bence Jones protein leads to formation of "amyloid-like" fibrils in vitro.

Motivated by the finding that the amino acid sequence of the Bence Jones protein BJP-DIA was identical to that of the main protein component of the amyloid fibrils obtained from the same patient with AL-amyloidosis, (Klafki, H.-W., Kratzin, H.-D., Pick, A.-I., Eckart, K., Karas, M. & Hilschmann, N. (1992) Biochemistry 31, 3265-3272.), we attempted to create "amyloid-like" fibrils from the Bence Jones protein in vitro, without addition of proteolytic enzymes. Reduction of BJP-DIA, solubilized in PBS, pH 7.4, overnight at 37 degrees C resulted in the formation of a precipitate which had affinity for the dye Congo red. Electron microscopy of negatively stained samples of the reduced protein revealed aggregates of linear unbranched fibrils. SDS-polyacrylamide gel electrophoresis demonstrated that the precipitate consisted almost exclusively of intact light chain molecules. This result makes it possible to deduce a molecular model of these amyloid fibrils generated in vitro.

Amyloid↗

Osteomesopyknosis: a benign familial disorder of bone.

A family with osteomesopyknosis is described. This condition is a rare benign bone dysplasia, characterized by patchy sclerosis predominantly involving the axial skeleton. The radiographic appearances may be confused with osteoblastic metastases, renal bone disease and other sclerosing bone disorders. As routine blood tests and isotope bone scans are normal in osteomesopyknosis, differentiation is readily achieved.

Aged↗

Centiles for adult head circumference.

Reference range for head circumference on the Tanner charts do not go beyond age 16. In this study the head circumference and heights of 354 adults in two British centres were measured. The centile charts constructed from these measurements show that adult head circumference is related to height. The mean head circumference of a male of average height is above the 97th centile for a 16 year old on the Tanner charts. The paediatric charts are therefore inappropriate for use in adult males.

Adolescent↗

Studies on human porin. VIII. Expression of "Porin 31HL" channels in the plasmalemma of the acute-lymphoblastic-leukemia cell line KM3 as revealed by light- and electron-microscopy.

First electron microscopy data on the expression of the VDAC "Porin 31HL" in the plasmalemma of a eucaryotic cell are presented. In a light and electron microscopic study we demonstrate the expression of the porin channel in the outer cell membrane of the pre-B lymphocyte type acute-lymphoblastic-leukemia cell line KM3. Monoclonal mouse anti-"Porin 31HL" antibodies were applied in indirect immunofluorescence or immunogold labelling experiments. The results confirm our early topological data on the expression of porin channels in the cytoplasmic membrane of different human cell types as revealed on the light microscopy level. Interestingly, only a pre-embedding immunoreaction approach was successful for gold particle labelling of the plasmalemma. This again is in agreement with our recent data on the accessibility of the acetylated N-terminal part of "Porin 31HL" molecules on the outer cell surface.

Antibodies, Monoclonal↗

Changes in body weight, body composition, and energy intake in women fed high- and low-fat diets.

The effects of a 20%-fat diet (LF) on total body weight, lean body weight, and adiposity were studied in 18 premenopausal women with body mass index (BMI) of 18-44. Subjects were fed a 37%-fat (HF) control diet for 4 wk followed by the LF diet for 20 wk. Total body weight, lean body weight, and fat weight were measured at the end of the HF and LF dietary periods by hydrostatic weighing. Despite adjustments in energy intake to maintain weight throughout the study, subjects exhibited a 2.8% decrease in total body weight (P less than 0.0006), an 11.3% decrease in fat weight (P less than 0.0001), and a 2.2% increase in lean body weight (P less than 0.0149) by the end of the LF period. Similar changes were observed in obese (BMI greater than 30) and non-obese women (BMI less than 30). By the end of the LF period, energy intake had increased significantly in comparison with the HF diet (119% of the HF intake, P less than 0.0001). Results could not be explained by changes in daily activity levels and suggest that macronutrient composition plays a role in energy requirements for weight maintenance.

Adipose Tissue↗

Expression of the genes for transthyretin, cystatin C and beta A4 amyloid precursor protein in sheep choroid plexus during development.

The levels of mRNA for transthyretin, cystatin C, and beta A4 amyloid precursor protein were measured in the choroid plexus of sheep embryos during different stages of development, using specific cDNA probes and Northern blot analysis. The 3 different mRNAs were detectable in the brain of very young embryos with a crown-rump length of 1 cm, corresponding to only a few days of gestation. The choroid plexus increased in weight very rapidly in the first half of gestation and much more slowly in the second half. The level of transthyretin mRNA in choroid plexus increased during the first half of gestation and stayed constant thereafter until birth, at a level of about 70% of that in choroid plexus of adult sheep. The proportion of mRNA for the proteinase inhibitor cystatin C in total RNA from choroid plexus increased throughout gestation to adult levels at birth. The concentration of the mRNA for beta A4 amyloid precursor protein in choroid plexus early in development was already as high as in adults and remained at this level throughout gestation. Messenger RNA for cystatin C or mRNA for beta A4 amyloid precursor protein was not detected in adult sheep liver.

Amyloid beta-Peptides↗

Gene expression in regenerating and acute-phase rat liver.

The integration of growth and the acute-phase response is investigated by comparing the mRNA levels in rat liver during acute inflammation with those after partial hepatectomy. Northern analysis is carried out for the mRNAs for thiostatin, alpha 2-macroglobulin, alpha 1-antitrypsin, inter-alpha-trypsin inhibitor subunit 1, haptoglobin, ceruloplasmin, transferrin, vitamin D-binding protein, alpha 1-acid glycoprotein, beta-fibrinogen, apolipoproteins A-IV and E, albumin, transthyretin, alpha 2-HS-glycoprotein, retinol-binding protein, beta-tubulin, c-myc protooncogene, glyceraldehyde-3-phosphate dehydrogenase, phosphoenolpyruvate carboxykinase, ornithine transcarbamylase, and alcohol dehydrogenase. The acute-phase response dominates during the first 18 h. Changes in mRNA levels related to growth of the liver become important thereafter, and the capacity for an acute-phase response of plasma protein synthesis becomes greatly reduced. The early increase in the level of ceruloplasmin mRNA observed during inflammation is abolished during regeneration, and that of vitamin D-binding protein mRNA is converted into a decrease. The mRNAs levels of glyceraldehyde-3-phosphate dehydrogenase increase, and those for phosphoenolpyruvate carboxykinase decrease during regeneration. Ornithine transcarbamylase mRNA levels are found to exhibit negative acute-phase regulation. The pattern of transcriptional regulation is similar during inflammation and regeneration.

Acute-Phase Proteins↗

The cDNA structure and expression analysis of the genes for the cysteine proteinase inhibitor cystatin C and for beta 2-microglobulin in rat brain.

Tissue patterns of gene expression were analyzed by measuring mRNA levels and incorporation of radioactive amino acids for cystatin C and beta 2-microglobulin, the two extracellular proteins in the brain with the highest ratio of concentration in cerebrospinal fluid over that in blood plasma. The primary structure of rat cystatin C mRNA from choroid plexus was determined by nucleotide sequencing of cloned cDNA and the tissue patterns of gene expression were analysed by RNA blot analysis and in situ hybridization. Cystatin C was found to be composed of 120 amino acids and to contain a potential site for N-linked glycosylation. The tissue with the highest cystatin C mRNA level was the choroid plexus of the brain. Cystatin C mRNA was also detected in lower levels in other areas of the brain, testis, epididymis, seminal vesicles, prostate, ovary, submandibular gland, and, in trace amounts, in liver. Choroid plexus pieces in culture secreted radioactive cystatin C when incubated with radioactive leucine. Rat beta 2-microglobulin cDNA was cloned and identified by nucleotide sequencing and comparison of the obtained sequence with that of mouse and human beta 2-microglobulin cDNA. Tissue levels of beta 2-microglobulin mRNA in the rat were measured by hybridization to rat beta 2-microglobulin cDNA. The highest levels of beta 2-microglobulin mRNA were observed in liver and choroid plexus. Other parts of the brain and testis contained lower levels of beta 2-microglobulin mRNA.

Amino Acid Sequence↗

Early regional variations in motoneuron numbers arise by differential proliferation in the chick embryo spinal cord.

Regional differences in the number of motoneurons in the spinal cord of the chick are thought to arise developmentally by region-specific cell death and cell migration. In this way, a numerically homogeneous motor column throughout the spinal cord is believed to be molded into the adult pattern. Region-specific differences in proliferation are not thought to play a significant role in this process. By counting motoneurons in serial sections throughout the rostral-caudal extent of the spinal cord on Embryonic Day 4 in the chick, we have found that the numerical variations in motoneurons in different spinal cord regions are already foreshadowed by this stage, which is before the onset of both cell death and the secondary migration of neurons out of the motor column. These results indicate that although nonproliferative events may contribute to the later regional variations in motoneuron numbers, the initial differences themselves are created early on by regionally specific proliferative events.

Animals↗

Synthesis of thiostatins (major acute-phase alpha 1 proteins) in different strains of Rattus norvegicus.

1. Two different thiostatin proteins were detected by crossed immunoelectrophoresis in plasma of the following inbred strains of Rattus norvegicus; Wistar, Sprague Dawley, Hooded Wistar, Lewis, Porton-Albino, ACI, Long Evans, and Katholiek, a mutant strain of the Brown Norway. 2. Only one thiostatin protein was detected using crossed immunoelectrophoresis in plasma from the Buffalo rat. 3. Comparison of partially purified thiostatins from Buffalo, Wistar, and Sprague Dawley rats in polyacrylamide gels showed that the thiostatin protein in Buffalo rat plasma corresponded to thiostatin 1 of the two thiostatins of the Wistar and Sprague Dawley rats. 4. Thiostatin 1 mRNA and thiostatin 2 mRNA, of approximately 1.7 kilobases, were both demonstrated in RNA from Buffalo rat liver.

Acute-Phase Proteins↗

The acute phase response in the rodent.

In the rodent, the general response to acute inflammation and tissue damage is characterized by a complex rearrangement in the pattern of concentrations of proteins in the plasma leading to an increase in the sedimentation rate of erythrocytes, an increase in leukocyte concentration in the bloodstream, and a decrease in the hematocrit. Body temperature changes only slightly or not at all. The reasons for the change in plasma concentrations of proteins are changes in their rates of synthesis in the liver. Degradation of plasma proteins is not affected. The details of the acute phase response evolved in the interaction of species with their environment. Therefore, it is not surprising to find differences in the details of the acute phase response among species. For example, alpha 2-macroglobulin is a strongly positive acute phase reactant in the rat, but not in the mouse; C-reactive protein is a strongly positive acute phase protein in the mouse, but is not found in the rat. An inducible acute phase cysteine proteinase inhibitor system, which has evolved from a primordial kininogen gene, has been observed so far only in the rat. The changes in the synthesis rates of acute phase proteins during inflammation are closely reflected by corresponding changes in intracellular mRNA levels. In the liver, the capacity to induce the acute phase pattern of synthesis and secretion of plasma proteins probably develops around birth. Changes in mRNA levels are brought about by changes in transcription rates or by changes in mRNA stability. Kinetics of mRNA changes during the acute phase response differ for individual proteins. The main signal compound for eliciting the acute phase response in liver seems to be interleukin-6/interferon-beta 2/hepatocyte stimulating factor, whereas interleukin-1 leads to typical acute phase changes in mRNA levels only for alpha 1-acid glycoprotein, albumin, and transthyretin. Plasma protein genes are expressed in various extrahepatic tissues, such as the choroid plexus, the yolk sac, the placenta, the seminal vesicles, and other sites. All these tissues are involved in maintaining protein homeostasis in associated extracellular compartments by synthesis and secretion of proteins. Synthesis and secretion of plasma proteins in paracompartmental organs other than the liver is not influenced by the acute phase stimuli.

Acute-Phase Proteins↗

Characterization of the receptor for glucagon-like peptide-1(7-36)amide on plasma membranes from rat insulinoma-derived cells by covalent cross-linking.

125I-Labelled glucagon-like peptide-1(7-36)amide was cross-linked to a specific binding protein in plasma membranes prepared from RINm5F rat insulinoma-derived cells using disuccinimidyl suberate. Consistent with the presence of a single class of binding site on the surface of intact cells, only a single radiolabelled band at Mr63,000 was identified by SDS-PAGE after solubilization of the ligand-binding protein complex. The band was not observed when 10nM glucagon-like peptide-1(7-36)amide was included in the binding assay, but 1 microM concentrations of glucagon-like peptide-1(1-36)amide, glucagon-like peptide-2 and glucagon did not decrease the intensity of labelling. No change in the mobility of the band was observed under reducing conditions, suggesting that the binding protein in the receptor is not attached to other subunits via disulphide bonds. In control incubations using plasma membranes from pig intestinal epithelial cells, which do not contain specific binding sites for glucagon-like peptide-1(7-36)amide, no cross-linked ligand-binding protein complex was observed.

Adenoma, Islet Cell↗

Accuracy and precision of analyses for total cholesterol as measured with the Reflotron cholesterol method.

We compared plasma cholesterol measurements made with the Boehringer Mannheim Reflotron reflectance photometric analyzer in 1298 capillary blood samples with measurements made in venous blood samples collected at the same time and analyzed in four standardized Lipid Research Clinics laboratories. The Reflotron measurements averaged 0.8% to 7.8% lower than the laboratory values. Correlations (r) between the two sets of measurements ranged from 0.92 to 0.96. In some samples, however, the Reflotron values differed from the laboratory values by greater than or equal to 12%; the cholesterol concentrations in these samples tended to be higher than in those for which better agreement was observed. The smaller negative biases were observed when test strips were used that were calibrated with reference to the Centers for Disease Control Reference Method for cholesterol. The agreement between sequential Reflotron values averaged less than or equal to 4.3%. There was an average difference of less than or equal to 1.0% between Reflotron measurements made in each of two sequential capillary blood samples taken from a single finger puncture.

Cholesterol↗