Changes in the activity of ‘active’ pyruvate dehydrogenase complex in the newborn of normal and diabetic rats

Summary At birth, hepatic active and dichloracetate-activated pyruvate dehydrogenase complex activities in the newborn of normal, mildly diabetic, and severely diabetic rats were similar. The active and dichloracetate-activated pyruvate dehydrogenase complex activities increased significantly during the first 2 and 6 postnatal h, respectively in the three groups of neonates (p<0.05). The greatest increase in both active and dichloroacetate-activated pyruvate dehydrogenase complex activity was observed in the neonates of mildly diabetic rats. Administration of glucose or insulin at birth to the newborn of normal rats caused a significant increase in the percentage of active pyruvate dehydrogenase complex activity within 1 h (p<0.01). Similar treatment caused no significant increases in the newborn of severely diabetic rats. The transient increases in active pyruvate dehydrogenase complex activity in the neonates of normal and diabetic rats were consistent with rapid disappearance of blood lactate during the first hours of postnatal life.     

Investigation of magnesium absorption in children using<Superscript>28</Superscript>Mg

Membrane-bound acid -glucosidase of human spleen was solubilized with either sodium cholate or Cutscum. The solubilized enzyme in type 1 (adult) Gaucher disease was less heat-stable than the normal enzyme, and when precipitated by ammonium sulphate it had a higher apparent molecular weight than the corresponding normal enzyme. The normal -glucosidase was activated by taurocholate, whereas the Gaucher enzyme was inhibited. The decrease in acid -glucosidase activity in Gaucher disease was associated with a profound deficiency of that form of the enzyme which bound to Concanavalin A. The results are consistent with faulty processing of newly synthesized acid -glucosidase in type 1 Gaucher disease.     

A case of anti-Jo1 myositis with pleural effusions and pericardial tamponade developing after exposure to a fermented Kombucha beverage

The pathogenesis of the idiopathic inflammatory myopathies has been postulated to be an environmental trigger causing the expression of the disease in a genetically predisposed patient. We report a case of anti-Jo1 antibody-positive myositis which was associated with pleural effusions, pericardial effusion with tamponade, and mechanics hands, probably related to the consumption of a fermented Kombucha beverage. Kombucha mushroom, a symbiosis of yeast and bacteria, is postulated to be the trigger for our patients disease owing to the proximity of his symptoms to the consumption of the Kombucha beverage.Abbreviations ALT Alanine transferase - AST Aspartate aminotransferase - CPK Creatinine phosphokinase     

Mechanical properties and imaging characteristics of remanufactured intravascular ultrasound catheters

Intravascular ultrasound (IVUS) as a routine device in interventional cardiology is handicapped by its high price. 19 factory-made, remanufactured IVUS catheters which consist of sterilized, used phased-array IVUS transducers inserted into a new catheter shaft were compared with 23 new IVUS catheters. 3 mechanical and 4 imaging characteristics were assessed on a 5 point scale (1 = unacceptable, 5 = excellent). Mechanical as well as imaging properties of remanufactured IVUS catheter were comparable to new catheters with excellent ratings for each of the evaluated characteristics in 38 to 94% of remanufactured catheters and 50 to 96% of new catheters. The initial failure rate for remanufactured IVUS catheters was 31.6% vs. 4.3% for new catheters (P < 0.05). Overall failure rate was 47.3% for remanufactured catheters vs. 8.7% for new catheters (P < 0.05). The failure was due to an electronic connecting problem occurring during mechanical stress to the IVUS catheter. In conclusion, remanufactured IVUS catheters offer mechanical and imaging characteristics which are comparable to new catheters. Improvements in the remanufacturing process to resolve the high rate of electronic connecting problems may make this a promising approach to substantially lower the price of IVUS catheters.     

Differences of cellular composition and adhesion molecule expression in “leukemic” as compared with “normal” human long-term bone marrow cultures

Summary Human long-term bone marrow cultures (HLTBMCs) were established with bone marrow samples collected from 15 patients with acute myeloid leukemia (AML) and compared with HLTBMCs from eight healthy volunteers. During 6 weeks of culture, the cellular composition of HLTBMCs was quantitatively studied. The cells of the HLTBMCs were divided into three main categories: fibroblasts, macrophages, and other cells (endothelial cells, hematopoietic cells and undefined cells). HLTBMCs derived from healthy volunteers demonstrated a very consistent development. The number of fibroblasts increased during culture and the number of macrophages decreased, resulting in a steady state after 3 weeks of culture. In contrast, HLTBMCs derived from patients with AML showed a strikingly different pattern of irregular development and a steady state was not reached under our conditions. The APAAP technique was used to demonstrate expression of adhesion molecules. VLA2, VLA5, VLA6, LFA1, Mac1, p150/95, ₂-chain, HCAM, ICAM1, NCAM, and VCAM1 were more expressed on normal as compared with leukemic bone marrow stromal cells, although this reached significance only for ₂-chain and NCAM. VLA1, 3, and 4 were expressed in a higher percentage on leukemic stroma (not significant). More expression was seen on normal as opposed to leukemic macrophages for the adhesion molecules tested, except for VLA5. The differences reached significance for the majority of molecules tested. It is concluded that striking differences exist in cellular composition and adhesion molecule expression between HLTBMCs from healthy individuals and those from patients with AML. This may have an impact on the pathogenesis of AML.     

Prevention of diabetic glomerulopathy in streptozotocin diabetic rats by insulin treatment

Summary Glomerular basement membrane thickness (GBMT) has been measured in streptozotocin diabetic rats treated with insulin. The study included 3 groups of 8 rats each: 1) a well-controlled group of diabetic rats under insulin treatment with a plasma glucose level reasonable close to normal values, 2) a poorly-controlled group also under insulin treatment with constant high plasma glucose values, and 3) an age and weight matched non-diabetic control group. After 6 months of diabetes, GBMT was measured applying an intercept method on 3 glomerular cross sections from each of the 24 animals. The measurements showed that mean GBMT was 132.2. nm in the non-diabetic control rats and 131.6 nm in the well-controlled diabetic rats. In the poorly-control-led group the mean GBMT was 140.4 nm, i.e. statistically significantly increased when compared to each of the two other groups, 2p=0.022 and 0.012 respectively.The results demonstrate that good blood glucose control in rats preserves normal GBMT.     

Gaucher disease (type 1): Physical and kinetic properties of liposomal and soluble ‘acid’ β-glucosidase

Acid -glucosidase of human spleen, from either normal controls or patients with type 1 (adult) Gaucher disease, was incorporated into phosphatidylcholine liposomes. The non-incorporated (soluble) Gaucherenzyme had a higher apparent molecular weight than had the corresponding control. Liposomal acid -glucosidase prepared from Gaucher-spleen was more thermostable than was the corresponding normal enzyme; it was also stimulated by acidic lipids to a much lesser extent. The results suggest that the genetic mutation in type 1 (adult) Gaucher disease has multiple effects on the glycoprotein form of acid -glucosidase.     

Systemic lupus erythematosus associated with recurrent lupus enteritis and peritonitis

We describe the case of a 41-year-old woman with systemic lupus erythematosus (SLE) who suffered from repeated reversible lupus enteritis characterized by marked edematous thickening of the small intestine. Ultrasonography (US) and computed tomography (CT) manifested as an accordion-like appearance and a target-like appearance, respectively. Resolution of gastrointestinal tract wall thickening was observed on follow-up US performed a week after the increase in predinosolone (PSL). We conclude that careful evaluation of sonographic and radiographic findings helps to establish the diagnosis of lupus enteritis.Abbreviations CT Computed tomography - FANA Fluorescent antinuclear antibody - GI Gastrointestinal - SLE Systemic lupus erythematosus - US Ultrasound     

Study of porcine and human isophane (NPH) insulins in normal subjects

Summary The plasma glucose, C-peptide and insulin responses to subcutaneously administered highly purified porcine, semi-synthetic and biosynthetic human isophane (NPH) insulin and diluting medium as control in normal male subjects were evaluated. Porcine and semi-synthetic human NPH insulins were administered at two dose levels of 0.15 and 0.30 U/kg body weight and biosynthetic human NPH at 0.15 U/kg body weight only. At the low dose level the three insulin preparations resulted in a similar maximal hypoglycaemic effect within 3–5 h after administration. However, over the remainder of the 11 h post-injection period, the plasma glucose level was lower after semi-synthetic human insulin. In contrast, at the 0.30 U/kg dose level, there was no difference in the early or late hypoglycaemic response between porcine and semi-synthetic human NPH insulins of equivalent pharmaceutical formulation. The clinical relevance of these findings needs further evaluation. The data suggest that for the intermediate-acting NPH insulin preparations, both the species of insulin, nature and quantity of the retarding protein and their subsequent interaction may determine their time-action characteristics.     

CD4+ and CD8+ subsets: Naive and memory cells in the peripheral blood of patients with systemic sclerosis

Summary Systemic Sclerosis (SSc; scleroderma) is associated with several immunological abnormalities, including altered proportion between lymphocyte subsets. Peripheral blood lymphocyte subsets from 25 patients with SSc were studied by two-colour flow cytometry using monoclonal antibodies against CD45RA and CD29 markers, which allow a dissection of CD4+ and CD8+ populations into naive and memory subsets. A decrease of the percentage of CD8+ (p<0.05) and of CD8+ CD29+ (p<0.001) cells was observed compared to that in 20 age and sex-matched controls. These abnormalities were not significantly associated with the extension of cutaneous disease or other clinical features of SSc nor with treatment, pattern of autoantibodies or HLA phenotype.     

Promoting healthy aging in cities: The Healthy Cities project in Europe

Health promotion is a growing field, spreading into various settings. Thispaper focuses on the city as a setting for health promotion and on theHealthy Cities project (HCP) of the Regional Office for Europe of theWorld Health Organization in particular. In the context of the HCP, variousactivities to promote the health of older citizens have been carried out inrecent years. These are illustrated in this paper through case studies.     

Haemodynamic and metabolic effects in diabetic ketoacidosis in rats of treatment with sodium bicarbonate or a mixture of sodium bicarbonate and sodium carbonate

Summary To examine factors determining the haemodynamic and metabolic responses to treatment of diabetic ketoacidosis with alkali, groups of anaesthetised and ventilated rats with either diabetic ketoacidosis (mean arterial pH 6.86–6.96, mean arterial blood pressure 63–67 mm Hg) or hypovolaemic shock due to blood withdrawal (mean pH_a 7.25–7.27, mean arterial blood pressure 36–41 mmHg) were treated with sodium chloride (saline), sodium bicarbonate or Carbicarb (equimolar bicarbonate plus carbonate). In the diabetic ketoacidosis series, treatment with either alkali resulted in deterioration of mean arterial blood pressure and substantial elevation of blood lactate, despite a significant rise in myocardial intracellular pH determined by ³¹P-magnetic resonance spectroscopy. These effects were accompanied by falling trends in the ratios of myocardial phosphocreatine and ATP to inorganic phosphate. Erythrocyte 2,3-bisphosphoglycerate was virtually absent in animals with diabetic ketoacidosis of this severity and duration. In contrast, in shock due to blood withdrawal, infusion of saline or either alkali was accompanied by a transient elevation of mean arterial blood pressure and no significant change in the already elevated blood lactate; erythrocyte 2,3-bisphosphoglycerate was normal in these animals. The effect of alkalinization in rats with severe diabetic ketoacidosis was consistent with myocardial hypoxia, due to the combination of very low initial erythrocyte 2,3-bisphosphoglycerate, alkali-exacerbated left shift of the haemoglobin-oxygen dissociation curve and artificial ventilation. No evidence was found for any beneficial effect of Carbicarb in either series of animals; Carbicarb and sodium bicarbonate could be deleterious in metabolic acidosis of more than short duration.Abbreviations DKA Diabetic ketoacidosis - 2,3-BPG 2,3-bisphosphoglycerate - pH_a arterial pH - pH_i intracellular pH - P_aCO₂ arterial PCO₂ - Pi inorganic phosphate - PCr phosphocreatine - MRS magnetic resonance spectroscopy - MABP mean arterial blood pressure     

Scoring prevalence and severity in gonarthritis: the suitability of the Kellgren & Lawrence scale

In clinical and epidemiological studies the Kellgren & Lawrence (K&L) scale was mainly used in the past to determine the radiological prevalence and severity of gonarthritis. The dependency of this scoring system on the presence of osteophytes has been criticised, and owing to its varying reliability, its appropriateness in cross-sectional and longitudinal studies is questioned. It was the purpose of this study to test whether the dependency of the K&L scale on the presence of osteophytes is reflected in a higher correlation of the overall score with a separate scoring of osteophytes than with other radiological features. Furthermore we compared the inter- and intrarater reliability with the reliability of individual radiological features. Knee radiographs of 40 patients were graded according to the K&L scale and for the presence and severity of five separate radiological features of osteoarthritis. We found a moderate correlation of the K&L scale with the radiological features of osteophytes, joint space narrowing and flattening of condyles. The intra- and inter-rater reliability of the K&L scale was higher than the reliability of the individual radiological features. The K&L scale seems not to be strongly dependent on the presence of osteophytes, i.e. there is not a higher correlation of the overall score with that feature than with other radiological features.     

Distinct cytoplasmic islet cell antibodies with different risks for Type 1 (insulin-dependent) diabetes mellitus

Summary The cytoplasmic islet cell antibody patterns of sera from islet cell antibody positive non-diabetic and diabetic endocrine autoimmune patients, and newly-diagnosed Type 1 (insulin-dependent) diabetic patients were characterised using four layer immunofluorescence with monoclonal antiproinsulin or anti-glucagon antibodies. Two distinct islet cell antibody types were identified. One gave a diffuse cytoplasmic staining in both Beta and Alpha cells (whole islet pattern), and was not affected by pre-incubation with rat brain homogenate. The other had a granular appearance with staining restricted predominantly to Beta cells (selective islet pattern) and was completely inhibited by pre-incubation with rat brain homogenate. Some sera appeared to have a mixed islet pattern, in which glucagon-positive cells gave a weaker cytoplasmic staining than proinsulin-positive cells. The granular selective pattern was found in sera from 19 (79%) of 24 non-diabetic endocrine autoimmune patients, in two (22%) endocrine autoimmune patients who developed Type 1 diabetes (p<0.0001 vs non-diabetic endocrine autoimmune patients), and in none of 19 newly-diagnosed diabetic patients. The whole islet pattern was found only in sera from patients who had, or who subsequently progressed to, Type 1 diabetes. This study has identified a novel islet cell antibody specificity and demonstrates that in islet cell antibody positive endocrine autoimmune patients, only islet cell antibodies which stain both Beta and Alpha cells are associated with progression to Type 1 diabetes.     

Morphometry versus densitometry — a comparison by use of casts of human coronary arteries

Geometric and densitometric methods for quantitative coronary arteriography have generally been compared by use of phantoms simulating arteries with circular lumina (Hole phantoms). We have used more adequate phantoms obtained by casting disease-free and atheromatous human coronary arteries. The phantoms, filled with contrast medium, were imaged digitally (1024 × 1024 × 10 matrix) under experimental conditions simulating routine coronary angiography. The angiographic diameters and the densitometric cross-sectional areas of 59 marked lumina were determined in single plane and orthogonal biplane raw images. Geometric calibration was performed by help of a 7F coronary catheter. For the densitometric calibration, we used a hole phantom attached to the image intensifier. The obtained luminal areas were compared to their true values determined previously by planimetry. The mean absolute error of single plane cross-sections obtained geometrically was 1.53 mm². Biplane imaging reduced it by a factor 2.4 to 0.64 mm². The corresponding mean absolute errors for densitometry were 0.56 mm² and 0.51mm². Single plane diameter measurements appear thus of very limited value for hemodynamic conclusions. In contrast, biplane geometric quantification was not markedly inferior to single plane and biplane densitometry.     

Enzyme replacement and beyond

During the last decade, enzyme replacement therapy for lysosomal storage diseases became a reality with the demonstration of its safety and effectiveness in type 1 Gaucher disease. Currently, enzyme replacement and several other potential therapeutic strategies are being developed for selected lysosomal storage diseases, including Fabry disease due to the deficient activity of -galactosidase A (-Gal A). The development and clinical evaluation of these new therapies require a stepwise process, each step being rigorously reviewed and approved by national or international regulatory agencies. For lethal disorders that affect small populations, such as many inherited metabolic diseases, this process can be accelerated by orphan drug and fast track regulations. As an example of the drug development process, the development of recombinant human -Gal A (r-hGal A) replacement for Fabry disease is presented, including the preclinical studies in the Fabry mouse model, and the clinical phase 1/2, phase 3, and phase 3 extension studies, which demonstrate the safety and efficacy of this new therapy.     

Studies on Mv red cells

Summary The properties of the M^v antigen, a low incidence receptor of the MNSs blood group system, were investigated by serological tests with protease treated red cells and inhibition assays with glycoproteins or peptides from normal and M^v erythrocytes. Our data demonstrate that the M^v receptor represents an allelomorphic form of the N antigen on the Ss sialoglycoprotein, rather than a variant of the M receptor on the MN sialoglycoprotein. Anti-M^v plus -N (serum Arm.) reacts with the N, N and M^v antigens, whereas anti-M^v (serum Arch.) is specifically directed against the latter receptor.Preliminary results of this work were presented at the 7th International Congress of the Society for Forensic Haematogenetics [10]     

Oral glucose tolerance and ambient temperature in non-diabetic subjects

Summary When either a 960-kcal, 140-g carbohydrate meal, or a 75-g glucose load was ingested by non-diabetic Caucasians, the 2-h venous plasma glucose concentration was higher by 0.82 and 1.25 mmol/l, respectively, if the ambient temperature was 33 °C rather than 23 °C. It is likely that this is a result of relative arterialisation of the venous blood. Even at 23 °C room temperature, use of the hot hand technique to obtain arterialised venous blood increases post-load glucose levels in contralateral antecubital veins. If these observations apply to those acclimatised to the heat, they could affect the diagnosis of both diabetes and impaired glucose tolerance in the tropics.     

Women's intra-gender relationships and ‘disciplinary aggression’ in an Australian Aboriginal community

Based on research conducted in an Aboriginal community in the north of Australia, this paper explores acts of aggression between older and younger women, specifically those in a child/caretaker relationship. An examination of 24 cases of disciplinary aggression, most between consanguineally related women, raises the question of whether abuse is really an appropriate label for these acts. In attempting to understand how Aboriginal meanings of relationship and aggression reverberate in women's experiences of their relationships, their aggression, and their victimization, the author argues that aggression in these instances can be interpreted as a kind of nurturance and that hurt is perceived as a potential of women's interaction. The paper concludes with an assessment of the implications of this perception for the construction and experience of women's intergenerational relationships.     

Contractile activity and prostacyclin generation in isolated coronary arteries from diabetic dogs

Summary The present study was aimed at determining the generation of prostacyclin (PGI₂)-like-material in coronary arteries from normal and diabetic (pancreatectomized) dogs as well as the contractile responses to prostacyclin of preparations from normal, diabetic and insulin-treated diabetic animals. PGI₂ produced a dose-dependent relaxation of coronary arteries from normal dogs. In contrast, those from diabetic animals were not relaxed; indeed, at low concentrations PGI₂ failed to evoke any effect but at higher ones it induced a distinct contraction. In arteries from diabetic animals treated with insulin, PGI₂ induced a biphasic contractile effect, which lay between that of normal controls and untreated diabetics. In addition the basal generation of PGI₂-like-material by coronary arteries was significantly higher in the diabetic (141±0.2 pg/mg, mean±SEM) than in normal dogs (59±0.2 pg/mg). The present experiments demonstrate that the generation of PGI₂-like-substance is significantly increased in coronary arteries from diabetic dogs, but the same vessels are unable to respond to added authentic PGI₂ with relaxation; on the contrary they react with a distinct positive contractile response.