Contribution of DRB1*04 variants to predisposition to or protection from insulin dependent diabetes mellitus is independent of dq

Certain DQ alpha/beta dimeric molecules have been shown to play a major role in determining susceptibility or resistance to IDDM. Whether or not predisposition associated with DR4 haplotypes is exclusively due to linkage to DQB1*0302 and DQA1*0301 alleles is still a controversial issue. A modifying effect of certain DRB1*04 subtypes on the susceptibility encoded by DQ alleles is possible, since not all DRB1*04-DQB1*0302 haplotypes are associated with the disease. The distribution of DRB1*04 subtypes was analysed in 240 DR4-positive Caucasian IDDM patients and 110 DR4-positive healthy controls using allele-specific hybridization after genomic amplification. Although an important contribution to IDDM predisposition was encoded by the DQB1*0302 allele which was found in the majority of patients (94.2% vs 64.7% in controls, Odd's ratio OR = 8.8, P < 0.0001), differences between DRB1*04 variants persisted after the effect of the DQB1 locus was removed by matching patients and controls for DQB1*0302. Thus, the DRB1*0402 allele conferred a strong IDDM-predisposing effect (OR = 3.1, P < 0.02) which was highly significant in the absence of DR3 on the second haplotype (OR = 5.6, P < 0.0001) but was not visible among DR3/4 heterozygote individuals. Conversely, the DRB1*0404 allele conferred a strong protective effect (OR = 0.26, P < 0.0001) which was dominant even in the presence of the associated high risk DR3 haplotype (OR = 0.21, P < 0.03). These data indicate that DQ molecules are not the sole contributors to the DR4-associated IDDM predisposition, and that peculiar DR4 subtypes play a significant role in susceptibility to or protection from the disease. DRB1*0402 differs from DRB1*0404 by only two acidic residues at positions 70 and 71 within the peptide binding groove, instead of amide and basic amino acids. This might induce changes of peptide binding specificity that correlate with the genetic linkage of IDDM predisposition.     

Systematic screening for diabetic eye disease in insulin dependent diabetes

It has been demonstrated that alveolar macrophages (AM) are permissive for human immunodeficiency virus (HIV-1) after in vitro infection. However, data concerning in vivo infection of AM by HIV-1 still conflict. Therefore, we investigated AM collected by bronchoalveolar lavage from 15 HIV-1-infected patients. HIV-1 p24 and Gp120 antigens and viral RNA were not detected by immunocytochemistry and in situ hybridization, respectively, using 35S-labeled 3 kb Pol-Env, 0.350 kb Gag, and 0.150 kb U5 LTR cRNA probes. In contrast, when using polymerase chain reaction on DNA extracted from purified AM, HIV-1 DNA was detected in the seven patients tested. After short-term culture of alveolar cells from three HIV-1-infected patients and in vitro stimulation with granulocyte/macrophage colony-stimulating factor (GM-CSF) and tumor necrosis factor-alpha (TNF-alpha), HIV-1 replication was observed in most of the AM. These results demonstrate that AM are latently infected by HIV-1 in vivo but are not a site for viral replication. In contrast, HIV-1 replication occurs when AM are withdrawn from their local environment, enhanced by GM-CSF and TNF-alpha stimulation. This suggests either a negative control or an inadequate stimulation of HIV-1 replication in the alveolar environment.     

The use of the Micral-Test strip to identify the presence of microalbuminuria in people with insulin dependent diabetes mellitus (IDDM) participating in the EUCLID study

In IDDM, microalbuminuria (urinary albumin excretion rate (AER) of 20-200 micrograms/min) is a predictor of persistent proteinuria and diabetic nephropathy. Early intervention may prevent or reduce the rate of progression of renal complications. The Micral-Test strip can be used to establish a semi-quantitative estimate of AER. We assessed the field performance of the Micral-Test strip in detecting microalbuminuria in the EUCLID study, an European wide, 18 centre study of 530 IDDM participants, aged 20 to 59 years. People with macroalbuminuria were excluded. On entry, all participants had albumin concentrations from two overnight urine collections measured by a central laboratory, and the corresponding Micral-Test performed on the two collections locally. a cut off of > or = mg/l albumin from the first Micral-Test, to detect a centrally measured albumin concentration > or = 20 mg/l, yielded 29 (5.8%) false negative results and 58 (11.6%) false positive results (sensitivity 70%, specificity 87%). The mean AER, from two collections, was compared with the corresponding 'pooled' Micral-Test results (mean of the two readings). Receiver Operating Characteristic (ROC) curves were used to assess if there was a suitable 'pooled' Micral-Test result for screening microalbuminuria. A 'pooled' Micral-Test result (> or = 15 mg/l) was used to detect mean AER > or = 20 micrograms/min (sensitivity 78%, specificity 77%). This 'pooled cut-off' had already been used for screening on to the study and led to an over-estimate (154 vs. 77) of the true number of microalbuminuric participants on the study. In conclusion, our findings suggest that the Micral-Test strip is not an effective screening tool for microalbuminuria, using the 'pooled' result from two measurements did not improve the sensitivity of the test.     

Psychosocial adjustment to and control of diabetes mellitus: Differences by disease type and treatment.

Studied the effects of clinically different types of diabetes and different treatment regimes on the psychosocial adjustment to the disease of 56 Ss with insulin dependent diabetes and 372 noninsulin dependent diabetics. The reliability and validity of the Diabetes Educational Profile, developed for assessment in this study, were examined and supported. Diabetes control was related to disease type and treatment. Psychosocial adjustment was related to disease type and treatment, and control was related to psychosocial adjustment. In assessing the clinical status of diabetics, any evaluation that includes psychosocial adjustment or diabetes control must consider the disease type and treatment. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

More uniform diurnal blood glucose control and a reduction in daily insulin dosage on addition of glibenclamide to insulin in type 1 diabetes mellitus: role of enhanced insulin sensitivity

Combination therapy with insulin and sulphonylurea has gained acceptance in management of subjects with Type 2 (non-insulin-dependent) diabetes mellitus. However, its role in management of Type 1 (insulin-dependent) diabetes mellitus remains controversial. In this study, the effect of combination therapy with insulin and glibenclamide on metabolic control, daily insulin dosage, and insulin sensitivity was assessed in subjects with Type 1 diabetes mellitus. Ten men with Type 1 diabetes mellitus participated in a randomized, double-blind, crossover, clinical trial with three treatment regimens, namely (1) insulin alone, (2) insulin and placebo, (3) insulin and glibenclamide, each lasting 3 months. Combination therapy induced: (1) reduction in daily insulin dosage; (2) more uniform blood glucose control as reflected by a lower average 24 h blood glucose level, a smaller difference between mean preprandial and 2 h postprandial blood glucose concentrations, decreased 24 h urine glucose excretion, and a decline in number of hypoglycaemic events; (3) improved insulin sensitivity as expressed by more rapid plasma glucose disappearance rate, without a significant alteration in fasting plasma glucagon and 1h postprandial serum C-peptide levels; when compared with treatment with either insulin alone or with insulin and placebo. Therefore, it is apparent that the addition of glibenclamide to insulin reduces daily insulin dosage and renders a greater uniformity to diurnal blood glucose control, most probably secondary to enhancement of insulin sensitivity.     

The ''thrifty'' hypotheses: clinical and epidemiological significance for non-insulin-dependent diabetes mellitus and cardiovascular disease risk factors

Cell adhesion molecules (CAMs) must play a crucial role in both the initiation and signalling of axon-glial contact. However, the proteins that permit myelinating oligodendrocytes to recognize the axons that they ensheath in the developing CNS are unknown. By a subtractive cDNA library strategy, we have identified neurofascin as a powerful candidate for such a molecule. Neurofascin is strongly but transiently up-regulated in oligodendrocytes at the onset of myelinogenesis. Once oligodendrocytes have engaged their target axons the protein plays no further part, since the expression of the gene declines precipitously, in contrast to that of the major myelin component proteolipid protein, which remains elevated. After the initial surge of neurofascin expression in oligodendrocytes, there is a shift to a predominantly neuronal localization that persists into adulthood. Hence neurofascin in oligodendrocytes is unlikely to serve a function in the stabilization of the multilamellar sheath around the axon. The major neurofascin isoform of oligodendrocytes contains the third fibronectin type 3 (FNIII) repeat but lacks the mucin-like domain which supports the view that neurofascin isoforms are differentially expressed in the nervous system. Among the genes that are up-regulated during the terminal differentiation of the oligodendrocyte, neurofascin is unique in displaying a transient pattern of expression at the early stages of myelination. We propose that this CAM not only has a role in mediating axon recognition but also signals axonal contact through its links with the actin cytoskeleton.     

Plasma disappearance of glycated and non-glycated albumin in type 1 (insulin-dependent) diabetes mellitus: evidence for charge dependent alterations of the plasma to lymph pathway

The fractional plasma escape rates of glycated and non-glycated albumin have earlier been measured in groups of Type 1 (insulin-dependent) diabetic patients and control subjects. The escape of non-glycated albumin was similar in control subjects and normoalbuminuric patients, but elevated in patients with micro or macroalbuminuria. In all groups the escape rate of glycated albumin was lower than that of non-glycated albumin. Glycation increases the anionic charge of albumin. To assay for charge-dependent alterations of transport a selectivity index (non-glycated albumin/glycated albumin transport ratio) was determined from the disappearance data. The index was high in control subjects (1.021 +/- 0.0057 (SEM)). This reflects a mean difference between the two escape rates of 2.1% per hour (for comparison the mean of the fractional escape rate of non-glycated albumin of the normal control subjects was 4.7% per hour). The index was numerically even higher in normoalbuminuric patients (1.031 +/- 0.0047 (SEM)), but reached significantly lower levels in patients with microalbuminuria (1.013 +/- 0.0030 (SEM), p < 0.02). Patients with clinical nephropathy had very low levels indicating loss of selectivity (1.002 +/- 0.0068 (SEM), p < 0.001). This pattern accords well with measurements of renal clearance selectivity indices, suggesting a general, progressive deterioration of anionic perivascular barrier components in diabetic microangiopathy. The structural target for these changes is likely to be the glycosaminoglycans of the glomerular basal membrane and the interstitial matrix.     

Thermopower method for detecting the ferrite-austenite transition: applications to surface reaction and diffusion

Difference of the thermoelectromotive force (temf.) for ferrite and austenite. Measurement of the change of thermopower during the transition ferrite–austenite, as a method for determination of rate parameters and diffusion constants. Application on the carburization and decarburization of Fe and Fe-Ti alloys and on the carbon diffusion in ferrite and austenite.     

Cutis/subcutis thickness at insulin injection sites and localization of simulated insulin boluses in children with type 1 diabetes mellitus: need for individualization of injection technique?

The study aimed to describe the variations of cutis/subcutis thickness at insulin injection sites in children with Type 1 diabetes mellitus and to localize the tissue position of a simulated insulin bolus in order to evaluate the need for individualization of injection technique in children. Cutis/subcutis thickness was measured by ultrasound in 47 children (25 girls and 22 boys) without compression (CSCUT) and with compression (CSCT) of the skin at 11 insulin injection sites. Tissue deposition of insulin was measured by ultrasound of a simulated insulin bolus of 200 microl of sterile air injected by the patients using their usual technique and site. On the thigh, 44% of girls and 95% of boys had a CSCT of less than 8 mm at one of the measured sites, while 16% of girls and 50% of boys had a CSCT of less than 6 mm at one injection site on the thigh and buttock. Significant differences in cutis/subcutis thickness in the same anatomical region were shown. CSCT was up to 35% less than CSCUT. The air bolus injection was placed inappropriately by 19% of children (using 8 mm needles). Unawareness of the skin thickness at the injection sites may contribute to inappropriate deposition. We propose that regular ultrasound measurements of subcutis depth at insulin injection sites be taken. This will allow the injection technique to be individualized (vertical or at an angle of 45 degrees). More children would be able to use the simpler vertical technique if 6 mm needles were used where available, or if even shorter (4 mm) needles were produced.     

Preimplantation diagnosis for Huntington's disease (HD): clinical application and analysis of the HD expansion in affected embryos

Huntington's disease (HD) is an autosomal dominant disease characterized by motor disturbance, cognitive loss and psychiatric manifestations, starting between the fourth and the fifth decade, followed by death within 10-20 years of onset of the disease. The disease-causing mutation is an expansion of a CAG triplet repeat at the 5' coding end of the Huntington gene. We have developed a single-cell PCR assay for the HD gene in order to propose preimplantation genetic diagnosis (PGD) for the couples at risk. We present here our first results with our first nine PGD cycles and also discuss the behaviour of the disease-causing expansion in pre-implantation embryos.     

Prevalence and predictors of risk for type 2 diabetes mellitus and impaired glucose tolerance in polycystic ovary syndrome: a prospective, controlled study in 254 affected women

Small molecular weight calcium salts, if absorbed intact, could provide a nutritional source of calcium in subjects with impaired absorption of calcium by the saturable pathway. An understanding of the mechanism of absorption of calcium oxalate (as a representative salt) may be important nutritionally and therapeutically. The aim of the present study was to develop models to study absorption, distribution and retention of calcium and oxalate in rats as a basis for studying calcium oxalate absorption. Labeled compounds (45Ca and [14C]-oxalic acid) were administered to separate groups of rats orally (n = 8-11) or intravenously (n = 3-5) and blood was sampled for up to 240 min. Data were analyzed using SAAM/CONSAM. Calcium kinetics were fitted by a model with three compartments in the body and one absorption pathway from the intestine. By contrast, oxalic acid kinetics were fitted by two pools in the body and two absorption pathways from the intestine. Calcium and oxalic acid, therefore, demonstrate different absorption and distribution kinetics in rats.     

Plasma homocysteine is related to albumin excretion rate in patients with diabetes mellitus: a new link between diabetic nephropathy and cardiovascular disease?

The high risk of cardiovascular disease in patients with diabetes mellitus, particularly in those with nephropathy, is not completely explained by classical risk factors. A high plasma homocysteine concentration is an independent risk factor for cardiovascular disease but information on its association with diabetes is limited. Fasting homocysteine concentrations were measured in the plasma of 165 diabetic patients (75 with insulin-dependent [IDDM]; 90 with non-insulin-dependent diabetes [NIDDM]) and 56 non-diabetic control subjects. Other measurements included the prevalence of diabetic complications, glycaemic control, lipid and lipoprotein levels, vitamin status and renal function tests. Patients with NIDDM had higher homocysteine levels than control subjects, whereas IDDM patients did not (9.2 +/- 4.5 vs 7.7 +/- 2 micromol/l, p < 0.01; and 7.0 +/- 3 vs 7.4 +/- 2 micromol/l, NS). Univariate correlations and multiple regression analysis showed albumin excretion rate to be the parameter with the strongest independent association with homocysteine. Patients with both types of diabetes and nephropathy had higher plasma homocysteine concentrations than those without nephropathy. Increases of homocysteine in plasma were related to increases in the severity of the nephropathy. Fasting hyperhomocysteinaemia was considered as the mean of the plasma homocysteine for all control subjects (7.5 +/- 2.1 micromol/l) + 2 SD (cut-off = 11.7 micromol/l). Nephropathy was present in 80 % of diabetic patients with fasting hyperhomocysteinaemia. In conclusion, increases in fasting homocysteine in diabetic patients are associated with increased albumin excretion rate, especially in those with NIDDM, thus providing a potential new link between microalbuminuria, diabetic nephropathy and cardiovascular disease.     

Consumption of drugs in diabetes mellitus II. Utilization and loss of insulin: epidemiologic and socioeconomic implications. Study Group for Diabetes in Tarragona (GEDT)

BACKGROUND: With the aim of evaluating the real consumption on insulin an analysis of its loss with use in clinical practice was carried out. The influence of this loss was investigated in the calculations of prevalence of diabetes (DM) initiating from the consumption of medication, the presumable repercussion in public health costs and possible alternatives. METHODS: Revision and analysis of the recipients used by a group of 58 insulin treated diabetics was carried out during a mean period of one month. The theoretic consumption, real consumption and the mean loss per each injection according to visual accuracy and the system employed were evaluated. A deduction was made of the autonomy by storing of insulin. A previous calculation concerning the prevalence of DM in Tarragona (548,900 inhabitants) according to consumption was corrected and an economic estimation of the loss demonstrated over public health costs of insulin during 1991 was made. RESULTS: The mean dose prescribed was 39.7 IU/day supplied in 2.4 injections/patient/day. At 30 days (27-35) 310 recipients were evaluated (115 vials/195 boxes). The mean real dose consumed was 53.3 IU/day and the mean loss per injection was 5.6 (25.5% of all the insulin supplied, 4.5% as remnants at the bottom of the recipient). A greater loss was observed by injection a) in patients with reduced sight (6.4 +/- 7.3 IU/5.5 +/- 4.5; NS) and b) in the users of syringes with dead space (5.8 +/- 4.7) with respect to those using an injector insulin pen (4.4 +/- 2.9; p < 0.01). The autonomy by domiciliary storage of insulin was of 103.7 days/patient (prescribed doses) and 78.6 (real consumption). A total of 7 diabetics (12%) had unused expired recipients. The prevalence of insulin treated DM in Tarragona was estimated as around 4.3-4.8/1,000 (2,360-2,635 inhabitants). The expense of loss was 36 million pesetas/year; 6.4 as depreciated remnants of insulin in the bottom of recipients. CONCLUSIONS: There is a great loss of insulin in clinical practice which may be avoidable and which influences the public health costs for diabetes. An adequate educative strategy and system of injection independent of user ability would reduce the costs.     

Scoring method for assessing rate adaptive pacemakers: application to two different activity sensors

To optimize programming of rate adaptive pacemakers (RAPs), we explored a new mathematical method to assess the performance of RAPs during daily-life tests, using customized Windows-based software. By stepwise discriminant analysis and linear regression, this method allows calculation of the acceleration and deceleration capacity of pacemakers and their general behavior during effort and recovery phases. Twenty-three patients (10 females and 13 males; 68 +/- 8 years) with chronic atrial fibrillation and a slow ventricular response were evaluated. They randomly received an accelerometer-controlled VVIR Dash Intermedics pacemaker (10 patients) or a vibration piezoelectric-controlled VVIR Sensolog III Siemens pacemaker (13 patients). All patients underwent the same test protocol: 6 minutes walking, 1.5 minutes climbing stairs, 1.5 minutes descending stairs, and 0.5 minutes sit-ups. By definition, the pacemaker responsiveness slope was programmed so that the heart rate response of paced patients during the walking test corresponded best to that of healthy controls. The slope was left unchanged for the other tests. We considered four scores: an acceleration score (EA score), an effort rate score (ER score), a deceleration score (RD score), and a recovery rate score (RR score). Scores ranged from -10 (hypochronotropic behavior of the pacemaker) to +10 (hyperchronotropic behavior), based on daily-life tests of 15 healthy controls (7 females and 8 males, 65 +/- 9 years). A score of 0 represented exact concordance with healthy controls. During stair descent, the Sensolog III produced excessive acceleration (EA score = +2.9 +/- 1.1) compared to: (1) stair climbing (EA score = -4.0 +/- 1.9; P = 0.01, with the same pacemakers); and (2) the Dash (+1.8 +/- 1.9; P = 0.04) and healthy controls (P = 0.02). The sit-up tests revealed a hypochronotropic response of both pacemakers compared to healthy controls, with a larger difference for the Sensolog III (EA score = -2.0 +/- 5.8; P = 0.04; RD score = -6.8 +/- 3.8' P = 0.02). We conclude that activity-driven pacemakers can accommodate brief activities, except for isovolumetric exercise such as sit-ups. During daily activities, accelerometer-driven pacemakers seem to provide a heart rate resoibse closer to that of healthy controls. Our new mathematical analysis is a simple and reproducible method for evaluating and quantifying the efficacy of any sensor-driven pacemaker.     

A tracer interaction method for nonlinear pharmacokinetics analysis: application to evaluation of nonlinear elimination

A drug tracer is most commonly applied to get information about the pharmacokinetics (PK) of a drug that is not confounded by an endogenously produced drug or an unknown drug input. An equally important use of tracers that has not been fully recognized is their use in the study of nonlinear PK behavior. In the present study a system analysis is applied to examine the interaction between drug molecules characteristic and intrinsic to any nonlinear process which enables the nonlinearity to be identified and modeled using a drug tracer. The proposed Tracer Interaction Methodology (TIM) forms a general developmental framework for novel methods for examining nonlinear phenomena. Such methods are potentially much more sensitive and accurate than previous methods not exploiting the tracer principle. The methodology proposed is demonstrated in a simulation study and with real data in a specific implementation aimed at determining the Michaelis-Menten (MM) parameters of nonlinear drug elimination while accounting for drug distribution effects. The simulation study establishes that the TIM-based, MM parameter evaluation produces substantially more accurate parameter estimates than a nontracer (NT) conventional method. In test simulations the accuracy of the TIM was in many cases an order of magnitude better than the NT method without evidence of bias. The TIM-based, MM parameter estimation methodology proposed is ideally suitable for dynamic, non-steady-state conditions. Thus, it offers greater applicability and avoids the many problems specific to steady state evaluations previously proposed. TIM is demonstrated in an evaluation of the nonlinear elimination behavior of erythropoietin, a process that likely takes place via receptor-based endocytosis. Due to its high sensitivity, accuracy, and intrinsic nonlinearity the TIM may be suitable for in-vivo studies of receptor binding of the many biotechnology produced peptide drugs and endogenous compounds displaying receptor-mediated elimination.