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Biomedical subjects

G Reach

Publications and source records attributed to G Reach.

At least 55 records · Page 3Linked to original sources
PubMed1997

Extreme insulin resistance: clinical management by external subcutaneous insulin infusion.

Management of very high insulin requirements in rare extreme insulin resistance syndromes is difficult and poorly documented. We report a case of a type B insulin-resistant patient requiring approximately 10,000 units of insulin per day, i.e. beyond the possibilities of current insulin formulations and delivery devices. Only the Panomat C10 portable pump model (Disetronic) and U500 Humulin (Lilly) allowed the required rate of 400 units per hour to be attained only when the reservoir was changed twice daily and the site and catheter were changed once daily. Three months after discharge, the patient was in good general and local condition, but with only fair diabetes control (glycated haemoglobin 9.5%).

Adult↗
PubMed1997

Minimisation of microbial contamination for potential islet xenografts using specific pathogen-free pigs and a protected environment during tissue preparation.

One major risk of islet xenotransplantation is transmission of infections. We thus compared microbial contamination during preparation of islets from 4 pigs conventionally breeded and slaughtered or 8 specific pathogen free (SPF) pigs, and different environmental conditions during pancreas excision. Pancreas harvested in a slaughterhouse (for conventional pigs) or in a protected autopsy room (for SPF pigs) were soaked in betadine solution and submitted to enzymatic digestion with collagenase. Islets were purified on histopaque gradient with a COBE 2991 processor. For each step of the process, a 10 ml aliquot was harvested and microbial contamination was analysed. For all animals, contamination of livers, which were not soaked in betadine solution, was also examined. Analysis of livers from the 4 conventional pigs showed polymicrobial contaminations (1,122 +/- 841 CFU/mg) with several species of Staphylococcus, Streptococcus, Bacillus and Enterobacteriaceae. For these conventional pigs, soaking of pancreas in betadine solution and presence of antibiotics in all media decreased the pancreatic contamination compared to hepatic contamination, but were unable to suppress it, as transport solution and crude suspension obtained after the digestion step with collagenase showed persistent contamination (9.7 +/- 2.4 and 10.5 +/- 4 CFU/ml, respectively). After islet purification by histopaque gradient, no medium remained contaminated. During analysis of the 8 SPF pigs, no liver exhibited contamination. Analysis of medium from each preparation step showed complete absence of contamination for 7 pancreases. Only one contamination with Staphylococcus simulans was observed for one pancreas in transport solution (6 CFU/ml), and persisted in digestion medium (16 CFU/ml). Finally, all purified suspensions were completely sterile. In conclusion, breeding conditions of pig islet donors, and controlled environment for pancreas excision, considerably influence the risk of microbial contamination. In order to limit the risk, SPF pigs are a suitable and compulsory source of islets.

Animals↗
PubMed1996

Use of a subcutaneous glucose sensor to detect decreases in glucose concentration prior to observation in blood.

The development of a hypoglycemic alarm system using a subcutaneous glucose sensor implies that a decrease in blood glucose is rapidly followed by a decrease in the signal generated by the sensor. In a first set of experiments the linearity and the kinetics of the response of sensors implanted in the subcutaneous tissue of normal rats were investigated during a progressive increase in plasma glucose concentration: the sensitivities determined between 5 and 10 mM and between 10 and 15 mM were not significantly different, and a 5-10 min delay in the sensor's response was observed. In a second set of experiments, performed in diabetic rats, the kinetics of the decrease in subcutaneous glucose concentration following insulin administration was monitored during a decrease in plasma glucose level, from 15 to 3 mmol/L. During the 20 first min following insulin administration, the sensor monitored glucose concentration in subcutaneous tissue with no lag time. Subsequently, the decrease in the estimation of subcutaneous glucose concentration preceded that of plasma glucose. This phenomenon was not observed when the same sensors were investigated in vitro during a similar decrease in glucose concentration and may be due to a mechanism occurring in vivo, such as the effect of insulin on glucose transfer from the interstitial space to the cells surrounding the sensor. It reinforces the interest of the use of implantable glucose sensors as a part of a hypoglycemic alarm.

Scintillation Counting↗
PubMed1996

Xenografts of porcine islets immunoprotected in hollow fibres reduce the incidence of diabetes in non-obese diabetic mice.

Non-obese diabetic (NOD) mice develop an autoimmune disease with a long prodromal period and constitute a model for investigating the prevention of human insulin-dependent diabetes mellitus. Since insulin injected prophylactically has been shown to reduce incidence of diabetes in NOD mice, we tested a new strategy consisting of prophylactic xenografts of porcine pancreatic islets immunoprotected in semipermeable hollow fibres. Female NOD mice were transplanted twice (at 60 and 180 days of age) with islet-containing or empty fibres. Within the group grafted with protected islets, the incidence of diabetes was reduced (37 vs 75%; p < 0.01), the onset of disease was delayed (p < 0.02), and the severity of lymphocytic inflammation of endogenous islets was reduced (p < 0.02). When already diabetic mice were not taken into account for analysis, blood glucose level was slightly lower in those grafted with islet-containing fibres (p < 0.04). Graft function was also evidenced by HPLC separation of porcine insulin in NOD sera. Histological and perifusion studies of fibres retrieved from recipients confirmed immunoprotection. During co-transfer, T splenocytes from mice grafted with islet-containing fibres were able to reduce the capacity of T cells from diabetic donors to adoptively transfer the disease (p < 0.01). Antigens for islet-cell autoantibodies (ICA) in pancreata from both groups were compared by immunofluorescence with the same ICA-positive human sera to ensure that differences were due to antigen quantitative changes. These antigens, which could serve as an index of a possibly more extensive antigen beta-cell rest, were decreased (p < 0.01) in mice grafted with protected islets. Reduction of diabetes and insulitis following early islet transplantation may thus be due to generation of cellular mechanisms that actively suppress disease, and possibly in part to a decrease in antigens which make beta cells less vulnerable to autoimmune aggression. These effects can be obtained with xenogeneic islets protected in hollow fibres, thereby eliminating the need for immunosuppression. Based on the concept of prophylactic insulin therapy, this form of insulin administration offers a controlled means of delivering insulin to meet the physiological needs of recipients.

Aging↗
PubMed1996

Comparison between a rapid glycohaemoglobin (HbA1c) immunoassay and other indices of glycaemic control.

The aims of the study were to compare glycohaemoglobin (HbA1c) values measured by DCA (a benchtop analyzer primarily designed for within-clinic rapid HbA1c determination) to a reference HbA1c method and home blood glucose monitoring, and to explore the possibility of an uniform expression of data. A total of 103 blood samples and the corresponding mean capillary glucose values (4.4 +/- 1.2 tests/day) of the preceding 2 months were collected from 34 insulin-dependent diabetic adults. We measured the correlations and agreements using the residual plots method and regression equations between HbA1c measured by DCA and high-pressure liquid chromatography (HPLC), and between DCA and capillary glucose values. A highly significant correlation (r2 = 0.85, P < 0.001) and an acceptable agreement (97% of values within 2 SD of the mean difference of 0.9% +/- 0.4%) was found between DCA and HPLC values. The regression equation calculated on the first half of the cases was: DCA (%) = 0.72 HPLC (%) +1.38. Of DCA values expressed in HPLC terms using this equation 87% fell within a clinically acceptable confidence interval when compared with measured HPLC data. A significant correlation (r2 = 0.40, P < 0.01) was found between DCA and capillary glucose values, and the regression equation was: DCA (%) = 0.34 capillary glucose (mM) +4.44. Of glycaemic levels calculated from DCA values using this formula 82% fell within a clinically acceptable error range when compared with measured glycaemic values. We conclude that the three methods of assessment of diabetes control are well correlated and that it is possible, with a degree of precision acceptable for the clinical setting, to express all data in uniform units, e.g. mM of capillary glucose or percentage of HPLC-HbA1c, though a simple correspondence table based on our transfer equations may be clinically sufficient and more handy.

Adult↗
PubMed1996

Modification of the sensitivity of glucose sensor implanted into subcutaneous tissue.

The mechanism of reducing the glucose sensitivity of sensors implanted into the subcutaneous tissue of the normal rat was evaluated (n = 10) by comparing sensitivities observed in vitro and in vivo. In vivo sensitivity was significantly lower than that observed in vitro before implantation (p < 0.005). Most interestingly, in vitro sensitivity immediately after explanation did not differ from that in vivo and increased progressively during rinsing (p < 0.02 after 30 min). These results demonstrate that the reduction of in vivo sensitivity was not due to a local factor or factors but to a reversible alteration of the glucose sensor characteristics induced in vivo by some local factor(s). This suggests that modifications of the outer sensor membrane, the nature of which remains to be determined, may prevent this effect and resolve the problem.

Animals↗
PubMed1995

A method for obtaining monodispersed cells from isolated porcine islets of Langerhans.

Micro or macroencapsulation of islets of Langerhans have been proposed as a bioartificial pancreas. Encapsulation of dispersed single cells instead of porcine islets should improve the oxygenation of encapsulated tissue. The aim of this work was, therefore, to develop techniques for dissociating porcine islets and test cell viability and function. After islet isolation and purification, islets were dispersed into single cells with collagenase and DNAse in either an extracellular type ionic solution or a UW solution. After culture, islets or cells were perfused with Krebs buffer. Two consecutive stimulations from 2.8 mM to 20 mM glucose were performed. Viability of cells (trypan blue) was higher than 85% after dispersion in ES or UW solutions. Islets or dispersed cells responded similarly to both stimulations with a return to basal rate between stimulations. No difference was found between cell function cultured during 18 hours or 6 days. However, islet function was improved by a long period of culture. In conclusion, this study demonstrates that dissociated cells performed as well as native islets up to six days culture.

Analysis of Variance↗
PubMed1995

[Self-regulated closed-loop therapy of diabetes].

The development of an artificial pancreas, a system in which insulin delivery would be regulated continuously by concomitant glucose levels, has been for a long time considered as a dream by diabetologists and diabetic patients. Two approaches are possible. The first one, electromechanical, consists of a glucose sensor, a computer and an insulin pump. Its development may be hindered by the inadequate biocompatibility of the glucose sensor. This led to consider at first the use of a disposable glucose sensor as part of a continuous glucose monitoring system. The second approach aims at developing a bioartificial pancreas, in which isolated islets of Langerhans would be protected against immune rejection by an artificial membrane permeable to glucose and insulin but not so to factors responsible for immune rejection. Thus, it may become possible to transplant a large number of patients before the onset of diabetic complications with xenogeneic islets, without need for any immunosuppression. Promises and obstacles of these two approaches are described herein.

Artificial Organs↗
PubMed1994

Elimination of the acetaminophen interference in an implantable glucose sensor.

Acetaminophen has been one of the most serious electrochemical interferences to oxidase-based amperometric biosensors that measure H2O2. A study was carried out to investigate various polymer materials for their selectivity as the sensor inner membrane. A composite membrane of cellulose acetate and Nafion was found to eliminate acetaminophen and other electrochemical interferences effectively while at the same time maintaining reasonable diffusivity for hydrogen peroxide. The excellent in vivo performance of the sensor was attributed not only to significantly reduced steady-state sensitivity to acetaminophen but also to very slow acetaminophen response. These features, combined with rapid acetaminophen clearance pharmacokinetics, led to the decreased response as demonstrated in the rat.

Acetaminophen↗
PubMed1994

Reduction of acetaminophen interference in glucose sensors by a composite Nafion membrane: demonstration in rats and man.

Amperometric glucose sensors typically monitor the production of hydrogen peroxide generated in the course of the enzymatic oxidation of glucose. At the applied potential necessary to oxidize the peroxide produced, other species are also electroactive and contribute to the signal. Interference of ascorbate or urate has been effectively eliminated, but that resulting from the widely used analgesic acetaminophen is not. The aim of this work was to reduce this interference, which was found to be possible by introducing a membrane constructed of Nafion. We compared the in vitro sensitivity to acetaminophen of five Nafion sensors with that of five non-Nafion sensors with identical glucose sensitivity (2.0 +/- 0.4 vs 1.9 +/- 0.1 nA.mmol-1.l-1, NS): sensitivity to acetaminophen was 12.2 +/- 2.7 vs 30.8 +/- 6.3 nA.mmol-1.l-1, respectively (p < 0.05). These sensors were tested in rats by implanting in each animal one Nafion and one non-Nafion sensors. The in vivo sensitivity to glucose was similar (0.33 +/- 0.09 vs 0.30 +/- 0.05 nA.mmol-1.l-1, NS). The current generated by an acetaminophen infusion (plasma acetaminophen plateau = 140 +/- 10 mumol/l) was much decreased in the case of the Nafion sensor: 0.5 +/- 0.3 vs 2.0 +/- 0.7 nA, p < 0.05). Five Nafion sensors were implanted in the subcutaneous tissue of normal human volunteers who were given on oral dose of 500 mg acetaminophen.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetaminophen↗
PubMed1994

Development of a glucose sensor for glucose monitoring in man: the disposable implant concept.

This paper describes the issues related to glucose sensing in the framework of the concept of disposable implants. The possibility of providing a diabetic patient with a continuous access to his blood glucose concentration and of detecting nocturnal hypoglycaemia would be a major breakthrough in diabetes therapy. To this end, our laboratories have developed a miniaturized, subcutaneous glucose sensor for glucose monitoring. It is based on the enzymatic, amperometric detection of glucose. This glucose sensor has been extensively evaluated in rats, dogs and more recently in human volunteers. Under experimental conditions, a controlled increase in blood glucose concentration is followed by an increase in the current delivered by the sensor. It is then possible to transform this current into an estimation of the glucose concentration. Experiments in rats have shown that the glucose sensor functions for up to 10 days when implanted in the subcutaneous tissue. Experiments in conscious dogs have shown that it works in a subcutaneous tissue closer to the human one. More recently, the glucose sensor was investigated in non-diabetic volunteers. These studies have demonstrated that this subcutaneous glucose sensor is able to provide a reliable estimation of blood glucose concentration in man, making it suitable for blood glucose monitoring. Biocompatibility of the glucose sensor, and particularly long-term tolerance, remains to be demonstrated.

Animals↗
PubMed1994

Complement activation by alginate-polylysine microcapsules used for islet transplantation.

A foreign body reaction is frequently observed around implanted microcapsules of alginate-polylysine. Since complement activation can play a role in this reaction, we checked in vitro the ability of empty alginate-polylysine microcapsules to activate complement. Human serum was incubated with microcapsules, and complement activation was evaluated by two methods: the complement hemolytic activity (CH50) and the assay of the C3adesArg fragment. The occurrence of complement activation in the presence of microcapsules was suggested both by a CH50 decrease and by high C3adesArg levels despite C3adesArg adsorption to the capsule membrane. Capsule membrane protection against the cytotoxic effects of complement was also tested. No hemolysis occurred when microencapsulated sensitized sheep erythrocytes were incubated with activated complement. In conclusion, the microcapsule membrane can protect cells against activated complement fragments. Nevertheless, alginate-polylysine microcapsules do activate complement, and this effect must be considered for its use as an implant.

Alginates↗
PubMed1994

A one-step, operator-independent method for isolating islets of Langerhans from the porcine pancreas.

Large-scale isolation of islets of Langerhans is one of the major obstacles in islet transplantation. Until now, isolation methods relied on enzymatic digestion, the duration of which relies on a decision dictated by the operator's experience. This approach has always hindered development of an automated method. The aim of this study was to develop a one-step method based on complete digestion of the pancreas. The original aspect of the technique (derived from the Ricordi method) is use of the University of Wisconsin (UW) solution in the digestion medium and a continuous flow collagenase processing circuit with local cooling and rewarming to allow tissue digestion to proceed at 37 degrees C while settling of the cell suspension takes place at 4 degrees C. A stopcock system permits the alternate use of two settling chambers so that while one is in the circuit, the other can be removed for centrifugation, resuspension of the crude islet preparation in collagenase in free UW solution, and further purification in a density gradient system. Ten experiments were performed, and 545,750 +/- 48,670 purified pig islets were obtained per totally digested pancreas. Histological studies showed cell integrity. Insulin secretion in response to double glucose stimulation under perfusion conditions demonstrated the functional viability of the isolated islets. In conclusion, this one-step method makes it possible to obtain a high number of viable islets of Langerhans in the absence of any decision by an operator, and it should therefore provide basis for an automated method.

Adenosine↗
PubMed1994

[Artificial and bioartificial pancreas for the treatment of diabetes mellitus].

The development of an artificial pancreas, a system in which insulin delivery would be regulated continuously by concomitant glucose levels, has been for a long time considered as a dream by diabetologists and diabetic patients. Two approaches are possible. The first one, electromechanical, consists of a glucose sensor, a computer and an insulin pump. Its development may be hindered by the inadequate biocompatibility of the glucose sensor. This led to consider at first the use of a disposable glucose sensor as part of a continuous glucose monitoring system. The second approach aims at developing a bioartificial pancreas, in which isolated islets of Langerhans would be protected against immune rejection by an artificial membrane permeable to glucose and insulin but not so to factors responsible for immune rejection. Thus, it may become possible to transplant a large number of patients before the onset of diabetic complications with xenogeneic islets, without need for any immunosuppression. Promises and obstacles of these two approaches are described herein.

Animals↗
PubMed1994

Preliminary report on cell encapsulation in a hydrogel made of a biocompatible material, AN69, for the development of a bioartificial pancreas.

The occurrence of an inflammatory reaction represents the major obstacle to the development of any implantable system including micro and macroencapsulation. The purpose of this study was to describe an encapsulation method for cells using a membrane made of AN69, a copolymer of acrylonitrile which is considered as a reference in biocompatibility in the field of haemodialysis. The hydrogel of AN69 was obtained after a coagulation step at room temperature followed by a solvent/non-solvent (water) exchange phase. Microcapsules were obtained by co-extrusion of AN69 collodion and saline (with or without cells). The function of encapsulated cells was assessed in vitro, demonstrating cell survival after the microencapsulation procedure. These preliminary data are consistent with the potential interest for the development of the microencapsulation procedure aimed at realising a bioartificial pancreas.

Acrylic Resins↗