A genome-wide search for human non-insulin-dependent (type 2) diabetes genes reveals a major susceptibility locus on chromosome 2.

Non-insulin-dependent (type 2) diabetes mellitus (NIDDM) is a common disorder of middle-aged individuals characterized by high blood glucose levels which, if untreated, can cause serious medical complications and lead to early death. Genetic factors play an important role in determining susceptibility to this disorder. However, the number of genes involved, their chromosomal location and the magnitude of their effect on NIDDM susceptibility are unknown. We have screened the human genome for susceptibility genes for NIDDM using non-and quasi-parametric linkage analysis methods in a group of Mexican American affected sib pairs. One marker, D2S125, showed significant evidence of linkage to NIDDM and appears to be a major factor affecting the development of diabetes mellitus in Mexican Americans. We propose that this locus be designated NIDDM1.

An approach for identifying simple sequence repeat DNA polymorphisms near cloned cDNAs and genes. Linkage studies of the islet amyloid polypeptide/amylin and liver glycogen synthase genes and NIDDM.

Genetic factors contribute to the development of NIDDM, and genes involved in regulating pancreatic beta-cell function and insulin's effects on glucose metabolism are good candidates for being NIDDM susceptibility loci. However, testing candidate genes for linkage to NIDDM depends on the identification of highly informative DNA polymorphisms in or near the candidate locus. Here we describe an approach for identifying highly polymorphic markers near candidate genes that utilizes the emerging physical map of the human genome. A sequence-tagged site from the candidate gene is used to screen the Centre d'Etude du Polymorphisme Humain megabase-insert yeast artificial chromosome library, which contains information on the physical localization of >3,000 genetically mapped simple sequence repeat DNA polymorphisms. Thus, identification of a yeast artificial chromosome containing the candidate locus will in many instances also identify a physically linked simple sequence repeat DNA polymorphism that can be used as a marker for the candidate gene in linkage studies. We have used this approach to identify a marker for the islet amyloid polypeptide gene on chromosome 12. The physical mapping of this gene to a yeast artificial chromosome showed that it was in the same yeast artificial chromosome as the gene encoding liver glycogen synthase, another possible NIDDM susceptibility gene. Affected sib pair studies using a simple sequence repeat DNA polymorphism physically linked to the islet amyloid polypeptide and liver glycogen synthase genes showed no evidence for linkage with NIDDM, indicating that they are not major genes contributing to NIDDM susceptibility.

Comparison of circulating phagocyte oxidative activity measured by chemiluminescence in whole blood and isolated polymorphonuclear leukocytes.

The generation of reactive oxygen species (ROS) was measured by chemiluminescence (CL) in whole blood and isolated polymorphonuclear leukocytes (PMNLs) in 113 subjects. There were no differences in ROS production in males and females, smokers and non-smokers and no age dependency. A significant correlation was found between whole blood CL parameters and between parameters derived from isolated cells, but there was no correlation when these parameters were compared using isolated PMNLs and whole blood. Variations in serum lipid levels did not account for the variability of CL parameters in blood or PMNLs. Both methodologies had the same validity in demonstrating priming of phagocytes. Zymosan-induced CL was more sensitive in detecting distinct functional states of phagocytes than FMLP-induced CL.

Endothelin-1 does not prime polymorphonuclear leukocytes for enhanced production of reactive oxygen metabolites.

UNLABELLED: The in vitro effect of endothelin-1 (ET-1) on the capacity of polymorphonuclear leukocytes (PMNLs) to generate reactive oxygen species (ROS) was investigated. Human PMNLs were separated from healthy volunteers and preincubated for 10 min. at 37 degrees C with varying concentrations (10(-7)-10(-12) M) of ET-1. After subsequent stimulation with FMLP (10(-7) M) or opsonized zymosan (0.5 mg/ml) the intra- and extracellular generation of ROS was assessed by luminol-amplified chemiluminescence, superoxide radical (.O2-) and hydrogen peroxide (H2O2) production. RESULTS: ET-1 alone failed to stimulate ROS generation. Neither the capacity for extracellular generation of oxygen metabolites nor the production of ROS with an intracellular origin was changed after preincubation of PMNLs with ET-1. ET-1 did not cause a shift of the .O2-/H2O2 production ratio after stimulation of PMNLs with FMLP. These findings suggest that ET-1 in vitro does not prime human PMNLs for enhanced production of ROS.

[The concept of ambulatory therapy of proximal deep venous thrombosis]. / Das Konzept der ambulanten Therapie der proximalen tiefen Venenthrombose.

In recent studies outpatient treatment of proximal deep venous thrombosis with low-weight-molecular heparin is shown as safe and effective as the treatment within the hospital. The incidence of recurrent thromboembolism, major bleeding and mortality is not higher in an ambulatory setting than in the hospital. The comfort of the patient is higher, the costs can be reduced. The present review discusses the different conditions for ambulatory treatment of deep venous thrombosis. The question will be addressed which patients will probably be considered for outpatient treatment of deep vein thrombosis.

Recurrent isolated calf muscle thrombosis due to a venous aneurysm of the soleal muscle veins.

A rare case of venous aneurysm involving the soleal muscle vein in an 18-year-old woman is presented. The patients showed three episodes of ultrasonographically proven calf muscle thrombosis within 2 years. After a short course of low-molecular-weight heparin at a therapeutic dosage, complete thrombus recanalization was achieved. To prevent further thrombotic episodes, surgery including ligation and resection of the aneurysm was performed. At the 3-month follow-up study the patient had completely recovered.

[Interdisciplinary quality circle for osteoporosis. First results from the Regional Advisory Board for Osteoporosis for Saxony-Thuringia]. / Interdisziplinäre Qualitätszirkel für Osteoporose. Erste Erfahrungen aus dem Regionalen Expertenkreis Osteoporose von Sachsen-Thüringen.

The Regional Advisory Board Osteoporosis (REKO) for Saxony-Thuringia has established interdisciplinary quality circles in different districts with the goal to standardize the diagnosis of osteoporosis. Therefore, they developed a standardized program for general practitioners, gynecologists, internists and orthopedics. The documentation sheet covers 5 areas: Identification of the anamnestic osteoporosis risk with 7 standardized questions: If the patient reaches 3 or more out of 13 possible points, we assume he is at risk. 3 out of 5 clinical symptoms, 3 out of 6 x-ray symptoms and osteoporosis typical results of the bone density measurement in combination with the anamnesis give us a scale which allows us to classify each symptom for diagnostic purpose. The differential diagnostic laboratory program includes: Blood sedimentation rate, calcium, alkaline phosphatase, creatine, TSH basal and 25 OH vitamin D3 in the serum. To check effectiveness of the antiresorptive therapy, bone specific resorption markers are sometimes usable. The program will be implemented this year in all quality circles, evaluated and then defined in its final form. Among the participants of the quality circles, the program is already usable and offers a reliable basis for the therapy.

[Hyperuricemia and gout: diagnosis and therapy]. / Hyperurikämie und Gicht : Diagnostik und Therapie.

In our modern society hyperuricemia is one of the most frequent metabolism disturbances. So far, every fourth man and every tenth woman suffer from an asymptomatic or a symptomatic hyperuricemia named gout. Mostly, over nutrition and malnutrition as well as other secondary factors with a genetically determined renal secretion disturbance of uric acid lead to an increase of serum uric acid. By deposition of uric acid crystals in tissues with intermittent immunologic activation of inflammation cells a manifestation of gout can be seen. The clinical image of gout varies widely. It may manifest as acute or chronic arthritis, tophi on the skin, subcutaneous tissue and the skeletal system as well as urate nephropathy. To eliminate the consequences of hyperuricemia in the long term, apart from a thorough anamnesis of nutritional habits a general examination of metabolic parameters is necessary to exclude a metabolic syndrome and other causes for a secondarily caused hyperuricemia. As gout is very often primarily caused by a renal secretion disturbance of uric acid special diagnostics should be done. Basing on literature research and inclusion of experts opinions this article represents the therapeutically options in treatment of hyperuricemia and gout with their resulting side effects and contraindications.

[Epidemiology of hyperuricemia and gout]. / Epidemiologie der Hyperurikämie und Gicht.

Also in the GDR during the last 10 years the frequency of the disturbances of the purine metabolism increased. With a rate of prevalence of about 20% this anomaly of metabolism reaches the highest percentage of the metabolic disturbances. The increasing accumulation of the disturbances of the purine metabolism on account of complex examinations depends above all on the changed eating and drinking habits of the population. Also influences on body-weight, blood pressure and bodily activities are to be taken into consideration. Sex, age and genetic loads belong to the non-correctable factors of influence. The influences of methodical problems must particularly be taken into consideration in long-term studies. The accumulation of uric arthritis is still relatively small in the GDR in contrast to other industrial countries. However, in our county exact epidemiologic investigations are still necessary.

[Extraarticular diseases and findings in gout]. / Extraartikuläre Krankheiten und Befunde bei der Gicht.

Disturbances of the renal function, urolithiasis and hypertension are observed in a high percentage already in the asymptomatic stage of the primary hyperuricaemia. They determine above all the fate of the patient with gout. In primary uric arthritis over-accidentally frequently obesity, steatosis hepatis, hyperlipoproteinaemias, manifest diabetes mellitus and premature arteriosclerosis are found, without up to now proving a causal connection. Also these diseases often coin the clinical picture and demand particular attention in the diagnostics of patients with gout.

[Prevention and therapy of disorders of purine metabolism]. / Prophylaxe und Therapie der Störungen des Purinstoffwechsels.

To the most important prophylactic measures for the prevention of a disturbance of the purine metabolism belong the normalisation of the body-weight, a mixed diet without protein carriers containing much purine, a regular physical activity, a life without hectic conditions as well as without an excess of conflict situations and a moderate taking of alcohol. These recommendations are of particular importance for patients endangered by gout. In patients with hyperuricaemia they form the basic treatment for the prevention of a uric arthritis, a urate nephrolithiasis and a gouty nephropathy. The clinical and paraclinical results of the directives must be continuously tested with regard to the effectiveness. To the comprehensive prophylaxis also belong the preventive measures against an associated disturbance of an associated disturbance of metabolism or concomitant disease. An additional medicamentous therapy is necessary, when the success of the basic therapy is insufficient and there are already organ manifestations of a disturbance of the purine metabolism. The application of antihyperuraemic drugs corresponding to types should be performed by uricostatic drugs, uricosuric drugs and citric acid-citrate mixtures. The approach according to a 3-step-programme is most useful. In the treatment of the concomitant diseases certain preventive measures are to be taken into consideration. The therapy of an acute attack of gout is at present without any essential problems. Patients with disturbances of the purine metabolism need a permanent care by the physician.

[Modern differential-therapeutic aspects of the drug therapy of chronic gout]. / Moderne differentialtherapeutische Aspekte der medikamentösen Behandlung der chronischen Gicht.

The present paper adopts a definite attitude to the differentiated therapy of the disturbances of the uric acid metabolism. This demands an exacter subdivision of the kind of the metabolic disturbance (types Ia, Ib, IIa, IIb, and III). On the basis of this classification in types an individually adapted therapy is possible. It might form the prerequisite of a still more effective meeting of the nephrogenic complications of the gout and of the reduction of the side effects of the necessary permanent therapy.

[Incidence of metabolic disorders in patients with recurrent urinary calculi]. / Hüfigkeit metabolischer Störungen bei rezidivierenden Harnsteinbildnern.

Of 117 formers of relapsing calcium oxalate calculi 75.2% showed a hyperuricaemia, 64.1% a hyperlipoproteinaemia, 19.7% a diabetes mellitus, 44.4% a hypertension, 11% a hyperuricuria (greater than 1.0 g/a day) and 7.5% a hypercalciuria. In 30 formers of relapsing uric acid calculi we could establish a hyperuricaemia in 93.3%, a hyperlipoproteinaemia in 76.7%, a diabetes mellitus in 36.7%, a hypertension in 80%, a hyperuricuria in 11% and a hypercalciuria in 3.8%. In comparison to formers of calcium oxalate calculi patients with uric acid calculi had a significantly higher body-weight. The results of our examinations make clear that in relapsing formation of urinary calculi frequently a complex disturbance of the metabolism is present which in every case must be established and treated.

Characterization of the chemiluminescence response of polymorphonuclear leukocytes stimulated with urate crystals.

We studied the luminol-amplified chemiluminescence (CL) response of human polymorphonuclear leukocytes (PMNLs) stimulated with synthetic monosodium urate monohydrate (MSUM) crystals. Stimulation of PMNLs prepared from healthy controls with MSUM resulted in a bimodal CL response with two peaks after 1 min and 4-6 min, respectively. MSUM caused neutrophils to produce reactive oxygen species (ROS), which are released from the cells as well as ROS with an intracellular localization. Proportionally more of the CL activity generated during the first peak was of extracellular origin. The MSUM-induced light-generating mechanism was azide-sensitive, required superoxide anion and was strongly suppressed by horseradish peroxidase.

Beta-adrenergic modulation of FMLP- and zymosan-induced intracellular and extracellular oxidant production by polymorphonuclear leukocytes.

Evaluation of catecholamine modulation of PMNL extracellular and intracellular oxidant production may reflect beneficial and harmful effects of beta-adrenergic agonists in various disease states. We investigated the kinetics and potency of adrenaline-mediated inhibition of oxidant generation in FMLP- and zymosan-stimulated PMNLs. In FMLP-stimulated cells, the short-term burst of oxidant generation was inhibited by adrenaline in a dose-dependent fashion. Intra- and extracellular chemiluminescence and extracellular superoxide anion and hydrogen peroxide generation showed similar IC50 values for adrenaline (1.3-3.0 x 10(-8) M) indicating that both extracellular and intracellular events were inhibited with the same potency. In contrast, intracellular oxidant production evoked by the phagocytosis of zymosan was only minimally affected by 3 x 10(-5) -3 x 10(-12) M adrenaline. Extracellular inhibition of oxidant production was also apparent in zymosan-stimulated cells. In conclusion, adrenaline's ability to depress extracellular generation of oxygen metabolites while retaining prolonged intracellular oxidant production for phagocytosis supports its beneficial role as selectively targeted physiological protector.

[Oxidatively modified lipoproteins and their antibodies in patients with antiphospholipid syndromeand systemic lupus erythematosus]. / Oxidativ modifizierte Lipoproteine und deren Antikörper bei Patienten mit Antiphospholipidsyndrom und Systemischem Lupus erythematodes.

The antiphospholipid syndrome (APS) with its typical clinical manifestations of recurrent thrombosis and fetal loss is biochemically defined by the presence of circulating antiphospholipid antibodies (aPL). The disease pattern has raised special interest as a possible link between autoimmunity and atherosclerosis. aPL, oxidized low density lipoproteins (oxLDL), and antibodies to oxLDL (Anti-oxLDL) are suggested to play an important role in atherogenesis. In the present study we compared the serum levels of oxLDL and Anti-oxLDL in APS patients (20 subjects with primary APS; 14 subjects with secondary APS) and nonAPS subjects (24 phenotypically healthy controls samples and 12 patients with systemic lupus erythematosus [SLE]) and investigated associations of the above mentioned parameters with the intima-media thickness (IMT), a clinical surrogate parameter of atherosclerosis.SLE patients with and without APS showed significantly increased levels of Anti-oxLDL as compared to the controls group (p = 0.038 and p = 0.007, respectively). In contrast, oxLDL levels were not significantly different between the controls group and patients. The Anti-oxLDL levels correlated significantly with anticardiolipin (p = 0.002) and beta(2)-glycoprotein I antibodies (p < 0.048), both from IgG isotype. Only SLE patients without APS revealed a significantly elevated production of reactive oxygen species indicating an increased proatherogenic oxidative stress in the circulation (p < 0.002). In the patient groups, the circulating levels of oxLDL and Anti-oxLDL showed no association with atherosclerosis as estimated by IMT. In conclusion, our experimental data do not support the concept of oxidative stress-induced accelerated atherosclerosis in APS patients.