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[24] Preparation of Stable Radioiodinated Polypeptide Hormones and Proteins Using Polyacrylamide Gel Electrophoresis
This chapter discusses the preparation of stable radioiodinated polypeptide hormones and proteins using polyacrylamide gel electrophoresis. Radioactively labeled polypeptide hormones and proteins are widely used as tracers in radioimmunoassays and receptor studies. The peptide or protein is most easily labeled using iodination with 125I or 131I. The radioactive iodine is substituted in the tyrosin
Plasmapheresis in the initial treatment of insulin-dependent diabetes mellitus in children
Several factors indicate that autoimmune mechanisms may play a part in the aetiology of insulin-dependent diabetes mellitus. At the onset of the disease in 10 children (aged 11-16 years) plasmapheresis was performed four times over one to two weeks. Seventeen age-matched children with the same clinical features served as controls. The C-peptide concentrations at onset were the same in the two grou
Expression of major histocompatibility antigens on pancreatic islet cells
Insulin-independent diabetes mellitus is often accompanied by manifestations of autoimmunity and is frequently associated with certain HLA haplotypes, predominantly DR3 and DR4. Because the major histocompatibility antigens are important determinants of the immune response in various tissues, we have investigated their expression on the pancreatic islet cells. Human, mouse, or rat islets of Langer
Stable lodinated polypeptide hormones prepared by polyacrylamide gel electrophoresis
Iodination of several insulin and proinsulin preparations, human growth hormone and bovine pancreatic polypeptide was performed using H2O2 and lactoperoxidase or chloramine T. The iodination mixtures were fractionated by polyacrylamide gel electrophoresis at pH 9.15 in long gel rods followed by simple elution of the iodinated products from thin gel slices. With this method 125I tracers with long s
Trypan Blue as a marker of plasma membrane permeability in alloxan-treated mouse islet cells
Suspensions of pancreatic islet cells from noninbred ob/ob-mice were incubated with Trypan Blue. Microscope photometry showed that apparently viable cells excluded the dye completely, whereas the nuclei of nonviable cells accumulated Trypan Blue by a saturable process. The nucleus-to-medium dye gradient was more then 30∶1 in media containing 0.1% or less Trypan Blue. The apparent affinity constant
ISLET AUTOANTIBODIES IN HUMAN PANCREATIC TRANSPLANT RECIPIENTS
Effects of dextran-linked chloromercuribenzoic acid on insulin release from microdissected pancreatic islets
Insulin release in response to dextran-linked p-chloromercuribenzoic acid was studied in microdissected pancreatic islets of non-inbred ob/ob-mice. No contamination of the dextran-linked mercurial with free chloromercuribenzoic acid was detected before or after the incubation with islets. In comparison with free mercurial, of the same thiol-blocking activity, the dextran-linked compound had a weak
On the possible role of thiol groups in the insulin-releasing action of mercurials, organic disulfides, alkylating agents, and sulfonylureas
The thiol activity of pancreatic islets was spectrophotometrically assayed as the formation of 6-mercaptonicotinic acid from the organic disulfide, 6, 6’-dithiodinicotinic acid. Islets containing more than 90% β-cells were microdissected from non-inbred oblob-mice. Comparisons of intact with homogenized islets indicated that the organic disulfide penetrates relatively slowly into the β-cells. When
Effects of insulin secretagogues on phospholipid metabolism in pancreatic β-cells
The effect of insulin secretagogues on the incorporation of [32P]ortho-phosphate into phospholipids was studied in microdissected islets from obese-hyperglycemic mice. Increased 32P-labelling was observed after incubation for 60 min with 10 mM l-leucine, 10 mM l-arginine or 20 mM d-glucose. Most of the label occurred in the phosphatidyl inositol fraction. The effect of l-leucine was additive to th
Specificity of cyclic AMP potentiation of glucose-stimulated insulin release
The insulin-releasing activities of cyclic and non-cyclic nucleotides were studied with isolated pancreatic islets from obese-hyperglycemic mice. Substitution of cylidine, inosine, guanosine, or uridine for the adenosine moeity of cyclic 3ʹ,5ʹ-adenosine monophosphate was associated with a total loss of insulin-releasing ability. The 5ʹ-nucleotide triphosphate derivatives of these nucleotides were
Effects of neutral and dibasic amino acids on the in vitro release of insulin
The insulin-releasing ability of some neutral and di-basic amino acids was studied in an in vitro system using micro-dissected pancreatic islets with a high proportion of insulin-producing β-cells. The neutral amino acids l-alanine and α-aminoisobutyric acid had no substantial effects on basal and glucose-stimulated insulin release. In contrast, 5–20 mm of the dibasic amino acid l-arginine gradual
Specificity of leucine stimulation of insulin release
The ability of leucine to stimulate insulin release was studied in an in vitro system, using microdissected pancreatic islets with a high proportion of the insulin-producing β-cells. When tested in a glucose-free medium, l-leucine stimulated insulin release over a wide concentration range with maximum effect at about 20 mm. The stimulation of insulin release was additive to that elicited by 10 mm
Orthostatic hypotension and cardiovascular risk
Orthostatic hypotension (OH) is a cardinal sign of cardiovascular (CV) autonomic dysfunction as a result of autonomic nervous system failure to control the postural hemodynamic homeostasis. The proportion of individuals with OH increases with aging and chronic conditions, such as neurodegenerative diseases, hypertension, heart failure, diabetes, renal dysfunction, autoimmune diseases, and cancer.
Pulmonary blood volume index as a quantitative biomarker of haemodynamic congestion in hypertrophic cardiomyopathy
AIMS : The non-invasive assessment of left ventricular (LV) diastolic function and filling pressure in hypertrophic cardiomyopathy (HCM) is still an open issue. Pulmonary blood volume index (PBVI) by cardiovascular magnetic resonance (CMR) has been proposed as a quantitative biomarker of haemodynamic congestion. We aimed to assess the diagnostic accuracy of PBVI for left atrial pressure (LAP) esti
Cardiovascular biomarkers and echocardiographic findings at rest and during graded hypovolemic stress in women with recurrent vasovagal syncope
Introduction: Vasovagal reflex is the most common type of syncope but its etiology is not fully elucidated. Venous return and cardiac output are key in hemodynamic control. The aim of the study was to assess cardiovascular biomarkers and echocardiographic measures at rest and during hypovolemia in women with and without a history of vasovagal syncope. Methods: Fourteen women (aged 18-30) suffering
Prospective virome analyses in young children at increased genetic risk for type 1 diabetes
Viruses are implicated in autoimmune destruction of pancreatic islet β cells, which results in insulin deficiency and type 1 diabetes (T1D)1-4. Certain enteroviruses can infect β cells in vitro5, have been detected in the pancreatic islets of patients with T1D6 and have shown an association with T1D in meta-analyses4. However, establishing consistency in findings across studies has proven difficul
Absence of Islet Autoantibodies and Modestly Raised Glucose Values at Diabetes Diagnosis Should Lead to Testing for MODY : Lessons From a 5-Year Pediatric Swedish National Cohort Study
OBJECTIVE: Identifying maturity-onset diabetes of the young (MODY) in pediatric populations close to diabetes diagnosis is difficult. Misdiagnosis and unnecessary insulin treatment are common. We aimed to identify the discriminatory clinical features at diabetes diagnosis of patients with glucokinase (GCK), hepatocyte nuclear factor-1A (HNF1A), and HNF4A MODY in the pediatric population.RESEARCH D
A polar dinosaur feather assemblage from Australia
Exceptionally preserved Mesozoic feathered dinosaur fossils (including birds) are famous, but recognized from only very few localities worldwide, and are especially rare in the Southern Hemisphere. Here we report an assemblage of non-avian and avian dinosaur feathers from an Early Cretaceous polar (around 70°S) environment in what is now southeastern Australia. The recovered remains incorporate sm
Interval and Consecutive Round Breast Cancer after Digital Breast Tomosynthesis and Synthetic 2D Mammography versus Standard 2D Digital Mammography in BreastScreen Norway
Background: Screening that includes digital breast tomosynthesis (DBT) with two-dimensional (2D) synthetic mammography (SM) or standard 2D digital mammography (DM) results in detection of more breast cancers than does screening with DM alone. A decrease in interval breast cancer rates is anticipated but is not reported. Purpose: To compare rates and characteristics of (a) interval breast cancer in