Skin autofluorescence for the risk assessment of chronic complications in diabetes: a broad excitation range is sufficient.

Skin autofluorescence (AF) is becoming an accepted clinical method for assessing the risk of chronic complications in diabetes mellitus (DM). In this study, the role of the excitation wavelength in the recognition of increased risk of diabetes-related chronic complications was investigated. An Excitation Emission Matrix Scanner (EEMS) was used to perform noninvasive measurements in four age-matched groups of patients with type 1 and type 2 DM, with and without chronic complications, as well as in a control group (N=97 in total). AF was calculated for excitation wavelengths in the range 355 - 405 nm. Mean spectra were assessed per group. AF values in both type 1 and type 2 DM patients with complications were increased compared to the control subjects (p < 0:01); this ratio remained practically constant, independent of the excitation wavelength. No emission peaks were distinctive for specific patient groups. We conclude that in these groups, no characteristic fluorophores dictate the use of a specific wavelength or set of wavelengths. The results show the validity of applying a broad excitation wavelength range for risk assessment of chronic complications in diabetes.

Microbubble-enriched lavage fluid for treatment of experimental peritonitis.

BACKGROUND: Relaparotomies and closed postoperative peritoneal lavage (CPPL) are performed to treat persistent peritonitis. This experimental animal study compared open abdominal lavage with CPPL, and evaluated the potential of microbubble-enriched lavage fluids to improve the efficiency of CPPL and reduce clinical morbidity, mortality and cost. METHODS: Fluorescent polystyrene spheres were injected intraperitoneally into 22 male Wistar rats to simulate localized peritonitis. After 18 h the rats received open abdominal lavage and CPPL, with and without microbubbles. Microbubbles were obtained by adding ultrasound contrast agents to continuous ambulatory peritoneal dialysis fluid. RESULTS: Open abdominal lavage was 3.5 times more effective in particle removal than CPPL, owing to better fluid dynamics. The introduction of air-liquid interfaces in the form of microbubbles made CPPL up to 2.4 times more effective than lavage without bubbles. Best detachment results were obtained when microbubbles with a flexible surfactant shell and longer blood elimination half-life were used. CONCLUSION: Open abdominal and CPPL lavage techniques are not efficient beyond a certain duration and volume as they do not cause bacterial detachment from the peritoneal membrane. Using surface tension forces from microbubbles significantly enhanced polystyrene particle detachment. These findings may have great consequences for the treatment of patients with peritonitis.

Application of a user-friendly comprehensive circulatory model for estimation of hemodynamic and ventricular variables.

PURPOSE: Application of a comprehensive, user-friendly, digital computer circulatory model to estimate hemodynamic and ventricular variables. METHODS: The closed-loop lumped parameter circulatory model represents the circulation at the level of large vessels. A variable elastance model reproduces ventricular ejection. The circulatory model has been modified embedding an algorithm able to adjust the model parameters reproducing specific circulatory conditions. The algorithm reads input variables: heart rate, aortic pressure, cardiac output, and left atrial pressure. After a preliminary estimate of circulatory parameters and ventricular elastance, it adjusts the amount of circulating blood, the value of the systemic peripheral resistance, left ventricular elastance, and ventricular rest volume. Input variables and the corresponding calculated variables are recursively compared: the procedure is stopped if the difference between input and calculated variables is within the set tolerance. At the procedure end, the model produces an estimate of ventricular volumes and Emaxl along with systemic and pulmonary pressures (output variables). The procedure has been tested using 4 sets of experimental data including left ventricular assist device assistance. RESULTS: The algorithm allows the reproduction of the circulatory conditions defined by all input variable sets, giving as well an estimate of output variables. CONCLUSIONS: The algorithm permits application of the model in environments where the simplicity of use and velocity of execution are of primary importance. Due to its modular structure, the model can be modified adding new circulatory districts or changing the existing ones. The model could also be applied in educational applications.

Development of the isolated dual perfused rat liver model as an improved reperfusion model for transplantation research.

The Isolated Perfused Liver (IPL) model is a widely used and appreciated in vitro method to demonstrate liver viability and metabolism. Reperfusion is performed in a controlled setting, however, via the portal vein only. To study transplant related questions concerning bile and transport of bile, the in vitro Isolated dual Perfused Liver model is revisited. The IdPL is an in vitro reperfusion model, using both portal vein and hepatic artery. Livers from 12 Wistar rats were flushed with University of Wisconsin-organ preservation solution, procured and reperfused in either the conventional IPL-model (n = 6) or the new IdPL-model (n = 6). Liver injury, assessed by the release of aspartate amino transferase and lactate dehydrogenase, showed similar levels during both IPL and I dPL reperfusion, only alanine amino transferase showed an improvement. Cumulative bile production showed an improvement: 176.3 +/- 8.4 in the IdPL compared to 126.1 +/- 12.2 microg/g-liver in the IPL (p < 0.05). Clearance of phenol red (PR) and taurocholic acid (TC) remained similar. At 90 minutes reperfusion the PR clearance showed 0.11 +/- 0.01 and 0.11 +/- 0.02 mg/30min/g-liver and the TC clearance 1.01 +/- 0.10 and 1.01 +/- 0.07 micromol/ml/30min/g-liver in the IPL and IdPL, respectively. Increasing the reperfusion time beyond the normally used 90 minutes resulted in a significant increase in transaminases and LDH and a decrease in bile production, liver morphology remained intact and glycogen content was appropriate. In conclusion, the IdPL-model showed similar or better results than the IPL-model, but the liver could not endure an extended reperfusion time using the IdPL.

Determining the centre of pressure during walking and running using an instrumented treadmill.

In this paper, a new method of determining spatial and temporal gait parameters by using centre of pressure (CoP) data is presented. A treadmill is used which was developed to overcome limitations of regular methods for the analysis of spatio-temporal gait parameters and ground reaction forces during walking and running. The design of the treadmill is based on the use of force transducers underneath a separate left and right plate, which together form the treadmill walking surface. The results of test procedures and measurements show that accurate recordings of vertical ground reaction force can be obtained. These recordings enable a separate analysis of vertical ground reaction forces during double support phases in walking, and the analysis of changes in the centre of pressure (CoP) position during subsequent foot placements. From the CoP data, temporal gait parameters (e.g. duration of left/right support and swing phases) and spatial gait parameters (i.e. left/right step lengths and widths) can be derived.

Hypothermic machine perfusion of the liver and the critical balance between perfusion pressures and endothelial injury.

Hypothermic machine perfusion (HMP) provides better protection against cold ischemic injury than cold storage in marginal donor kidneys. Also, in liver transplantation a switch from static cold storage to HMP could be beneficial as it would allow longer preservation times and the use of marginal donors. A critical question concerning application of HMP in liver preservation is the crucial balance between perfusion pressure and occurrence of endothelial injury. Rat livers were cold-perfused for 24 hours to study perfusion pressures for both hepatic artery and portal vein. Cold storage served as control and was compared to HMP-preserved livers using a mean arterial perfusion pressure of 25 mm Hg and a portal perfusion pressure of 4 mm Hg (25% of normothermic liver circulation) and to HMP at 50 mm Hg and 8 mm Hg perfusion, respectively (50% of normothermic liver circulation). UW solution was enriched with 14.9 micromol/L propidium iodide (PI) to stain for dead cells and with an additional 13.5 micromol/L acridine orange to stain for viable hepatocytes. A low PI-positive cell count was found using HMP at 25% of normal circulation compared to cold storage. The PI count was high for the HMP group perfused at just 50% of normal circulation compared to HMP at 25% and compared to cold storage. In summary, for liver HMP, perfusion at 25% showed complete perfusion with minimal cellular injury. HMP using perfusion pressures of 25 mm Hg for the hepatic artery and 4 mm Hg for the portal vein is feasible without induction of endothelial injury.

Augmentation of abdominal organ perfusion during cardiopulmonary bypass with a novel intra-aortic pulsatile catheter pump.

BACKGROUND: Current pulsatile pumps for cardiopulmonary bypass (CPB) are far from satisfactory because of the poor pulsatility. This study was undertaken to examine the efficiency of a novel pulsatile catheter pump on pulsatility and its effect on abdominal organ perfusion during CPB. METHODS: Twelve pigs weighing 89+/-11 kg were randomly divided into a pulsatile group (n=6) and a non-pulsatile group (n=6). All animals had a CPB for 120 min, aorta clamped for 60 min, temperature down to 32 degrees C, and a perfusion flow of 60 ml/kg/min. In the pulsatile group, a 21 Fr intra-aortic pulsatile catheter, which was connected to a 40 mL membrane pump, was placed in the descending aorta and activated by a balloon pump driver during the first 90 minutes of CPB until aortic declamping. Hemodynamics, organ blood flow, body metabolism, and blood trauma were studied during experiments. RESULTS: Compared with the non-pulsatile group during CPB, the pulsatile group had a higher systolic blood pressure (p<0.01), higher mean arterial pressure (p<0.05), and higher blood flow to the superior mesenteric artery (p<0.05). The hemodynamic energy, indicated by the energy equivalent pressure (EEP) was higher in the gastrointestinal tract and kidney in the pulsatile group (p<0.01, p<0.01). Abdominal organ perfusion status, as indicated by SvO 2 in the inferior vena cava, was higher in the pulsatile group (p<0.05) 30 min after cessation of CPB. Hemolysis indicated by release of free hemoglobin during CPB was similar in the two groups. CONCLUSION: Applying the pulsatile catheter pump in the descending aorta is effective in supplying the pulsatile flow to the abdominal organs and results in improved abdominal organ perfusion during the ischemic phase of CPB.

Difficulties in the fixation of prostheses for voice rehabilitation after laryngectomy.

In most patients with advanced or recurrent laryngeal or hypopharyngeal cancer, total laryngectomy is indicated. This means the loss of three main functions: phonation; respiration; and the prevention of aspiration during deglutition. Laryngectomy patients have various options to restore phonation: an oesophageal voice; an electrolaryngeal voice; or a tracheo-oesophageal voice. In the last case a silicone rubber shunt valve is placed in the tracheo-oesophageal wall and phonation is generated when exhaled air is forced through the oesophagus and neopharynx. This method is widely applied in Western Europe. In this paper we review the literature on fixation problems with shunt valves, tracheostoma valves and heat and moisture exchange (HME) filters. Tracheo-oesophageal speech without a valve is not considered. Despite 22 years of experience with the implantation of tracheo-esophageal shunt valves and many improvements in the design, problems still remain, such as biofilm formation with subsequent leakage through the valve, the need for frequent and inconvenient replacements, fistula enlargement leading to leakage around the device and reduced fixation, and infections. The high cost of shunt valves is a drawback to their use worldwide. To enable hands-free speech, different types of tracheostoma valve have been developed. These valves are fixed to the skin or to the tracheostoma by means of an intra-tracheal device. An HME filter is used to protect the airway and maintain physiological balance. Such devices are only suitable for a selected group of patients as fixation to the skin or trachea can be a major problem. Speaking and coughing cause pressure increases, which often result in mucous leakage and disconnection of the valve and/or HME filter. Recommendations are made for future improvements in fixation techniques.

The influence of body mass index on the accumulation of advanced glycation end products in hemodialysis patients.

BACKGROUND/OBJECTIVES: The level of skin autofluorescence (AF) at a given moment is an independent predictor of mortality in hemodialysis (HD) patients. Skin AF is a measure of the accumulation of advanced glycation end products (AGEs). The aim of the study was to estimate the influence of nutrition on the 1-year increase of skin AF (ΔAF) in HD patients. SUBJECTS/METHODS: A total of 156 HD patients were enrolled in this study. Skin AF, body mass index (BMI), superoxide dismutase, myeloperoxidase, C-reactive protein, inter-cellular adhesion molecule-1, von Willebrand factor and heart-type fatty acid-binding protein were measured four times at intervals of approximately half a year. Data from the monthly routine blood analysis were also used. Daily calorie, protein and AGE intakes were assessed from food recordings over a period of 1 week. RESULTS: A J-shaped relation was found between baseline BMI and ΔAF (P=0.01). The lowest point of the J-shaped curve is found for BMI=24.3 kg/m(2). In the univariate analysis of the contributors to the 1-year ΔAF, we found that beside BMI=24.3 kg/m(2), AGE and calorie intakes, as well as myeloperoxidase and HD vintage, had a P <0.10. The sole independent predictor of the 1-year ΔAF was BMI=24.3 kg/m(2) (P=0.01). CONCLUSIONS: It appears that calorie, protein and AGE intakes hardly influence the 1-year ΔAF in HD patients. BMI of HD patients of around 24 kg/m(2) resulted in a lower 1-year ΔAF.

Corrigendum. The influence of body mass index on the accumulation of advanced glycation end products in hemodialysis patients.

Correction to: European Journal of Clinical Nutrition (2015) 69, 309­313; doi: 10.1038/ejcn.2014.261; published online 14 January 2015 Since the publication of this article, the authors have noticed that several of the author names were published incorrectly. The correct author names are listed above. The .html and online PDF versions have also been amended. The authors apologise for any inconvenience caused.

A novel tracheal tissue connector for fixation of laryngeal prostheses.

A tissue connector (TC), basically consisting of a ring that will be integrated into the trachea, is under development to study the fixation of laryngeal prostheses. Two experiments have been performed to test the TC in goats. In experiment 1, a polypropylene mesh was implanted around the trachea. The meshes were explanted after 6 and 12 weeks. In experiment 2, the actual TC consisted of two titanium rings (inner ring and outer ring) executed as quarter rings, fixed on each other, and a polypropylene mesh like a sandwich in between. The titanium inner ring was implanted between two tracheal rings thus penetrating the trachea with the mesh around the trachea and the fixed titanium outer ring on the outside of the trachea. The TCs were removed after 12 weeks. Experiment 1 showed that the mesh was entirely infiltrated by host tissue. Inflammatory cells and high vascularisation were observed in 3 of 4 implants. However, in experiment 2, the mesh was completely incorporated by mature connective tissue without inflammation reaction. At some areas, deposition of cartilage tissue was observed. In conclusion, the TC was firmly embedded in the trachea thus being appropriate for its intended use.

Biocompatibility of a novel tissue connector for fixation of tracheostoma valves and shunt valves.

Rehabilitation after laryngectomy often includes the use of a shunt valve and a tracheostoma valve to restore voice. To improve the fixation method of these valves, a new tissue connector has been developed, basically consisting of a ring that will be integrated into surrounding tracheal soft tissue. The valves can be placed in the ring. To test the principle of the tissue connector, a prototype consisting of a subcutaneous polypropylene mesh and a percutaneous titanium stylus was implanted into the backskin of 10 rats by a two-stage surgical procedure. We reasoned that if a firm connection can be realized with the skin, a firm connection with the trachea will also be possible. The subcutaneous part was implanted first, followed by the percutaneous part after 6 weeks. The complete tissue connector with surrounding tissue was removed 8 weeks later and examined histologically. The principle of the new tissue connector proved to be effective: hardly any epithelial downgrowth appeared, and adhesion of soft tissue was demonstrated. No infection or severe inflammation reaction was detected. The tissue connector seems appropriate for its intended use.

Animal models for tracheal research.

Tracheal research covers two main areas of interest: tracheal reconstruction and tracheal fixation. Tracheal reconstructions are aimed at rearranging or replacing parts of the tracheal tissue using implantation and transplantation techniques. The indications for tracheal reconstruction are numerous: obstructing tracheal tumors, trauma, post-intubation tissue reactions, etc. Although in the past years much progress has been made, none of the new developed techniques have resulted in clinical application at large scale. Tissue engineering is believed to be the technique to provide a solution for reconstruction of tracheal defects. Although developing functional tracheal tissue from different cultured cell types is still a challenge. Tracheal fixation research is relatively new in the field and concentrates on solving fixation-related problems for laryngectomized patients. In prosthetic voice rehabilitation tracheo-esophageal silicon rubber speech valves and tracheostoma valves are used. This is often accompanied by many complications. The animal models used for tracheal research vary widely and in most publications proper scientific arguments for animal selection are never mentioned. It showed that the choice on animal models is a multi-factorial process in which non-scientific arguments tend to play a key role. The aim of this study is to provide biomaterials scientists with information about tracheal research and the animal models used.

Transneuronal viral labelling of rat heart left ventricle controlling pathways.

Retrograde transneuronal viral labelling and immunocytochemical methods were used for revealing neuronal networks controlling the left ventricle myocardium of the rat heart. After injections of 1 microliter pseudorabies virus solution (3 x 10(6) PFU ml-1) into the left ventricle, infected orthosympathetic preganglionic cells were found in the intermediolateral cell groups of the first 6 thoracic spinal segments. Preganglionic parasympathetic neurones were seen both in the nucleus ambiguus/retro-ambiguus area and the dorsal motor vagus nucleus. Large numbers of infected projecting interneurones were found in the rostral, caudal and medial parts of the ventral medulla oblongata, the Kölliker-Fuse nucleus and catecholaminergic cell group A5 and in the paraventricular hypothalamic nucleus.

Evaluation of a porcine internal mammary artery (No-React II) as a small-diameter conduit.

BACKGROUND: The patency of biologic small-diameter vascular grafts in the aortocoronary position is still unsatisfactory. Most of the studies suggest that xenografts are to be avoided as an aortocoronary bypass. METHODS: The porcine internal mammary artery treated by the No-React II procedure was developed for use as an alternative coronary artery bypass conduit. The attempt of this study was to evaluate the patency and histologic changes of the porcine internal mammary artery in animals. Five calves underwent coronary artery bypass grafting with a porcine internal mammary artery graft to the right coronary artery. After euthanasia of the animals 103 days later, the samples of these grafts were studied morphologically for patency, structural changes, calcifications, and inflammatory and immunologic response. RESULTS: One animal died during the procedure as result of acute thrombosis of the porcine internal mammary artery graft. In the other 4 animals all grafts became occluded. In the histologic sections of the grafts we noted multiple calcifications and a host-graft immunologic reaction (severe chronic rejection). CONCLUSIONS: The present study demonstrates a very poor experience with the porcine internal mammary artery (No-React II) conduit. We do not recommend this prosthesis for clinical use in humans.

Development of an animal model of selective coronary atherosclerosis.

BACKGROUND: Atherosclerosis causes over 40% of all deaths in the USA and Western Europe. Although several hypotheses have been proposed, the etiology and pathogenesis of the atherosclerosis remain unknown. OBJECTIVE: To develop a model of selective coronary atherosclerosis in pigs. DESIGN: An animal model of selective coronary atherosclerosis was developed by combining a guide-wire-induced endothelial injury and cholesterol-enriched diet. METHODS: Twelve pigs were subjected to guide-wire-induced injury to endothelium of left anterior descending (LAD) coronary artery. Six animals (control group) were fed a standard pig food; the remaining six animals (cholesterol group) were fed a 6%-cholesterol-enriched diet. Three animals from the control group were killed immediately after the endothelial injury (acute control group). The other three animals in the control group (chronic control group) and all animals in the cholesterol-fed group were killed 4 weeks after the injury. RESULTS: The endothelial surface and the media of the left circumflex coronary artery LCX in all animals were intact. Long eccentric areas of endothelial injury were found in the LAD coronary arteries of animals in the acute control group. Numerous fibrous atherosclerotic plaques in LAD coronary arteries were found in animals in the chronic control group as well as in animals in the cholesterol-fed group, but were highly pronounced in animals in the last group. No accumulation of lipids was found in the plaques of animals in both groups. CONCLUSIONS: Administration of a 6%-cholesterol diet for 6 weeks is not sufficient to cause coronary atherosclerosis in pigs. Selective coronary atherosclerosis can be induced within 4 weeks with the same diet when the blood vessel has been injured with a guide wire.

Analysis of the mechanical behavior of the Nijdam voice prosthesis.

The valveless Nijdam prosthesis is a new voice prosthesis for laryngectomized patients using tracheoesophageal speech. An "umbrella-like hat" covers the esophageal side of the tracheoesophageal fistula and is deformed during speech by air pressure. To decrease pressure loss during speech, a good understanding of the mechanical behavior is essential. In the present study, the Finite Element Method (FEM), used in engineering to analyze the mechanical behavior of complex structures, was applied to analyze eight possible improvements of the Nijdam prosthesis. This study found that, during speech, deformation of hat and soft tissue occur. Distinct differences in the hat's deformation of the eight models also were found. It is concluded that complex structures like the Nijdam prosthesis can be analyzed by FEM. An optimal model was found to decrease pressure loss while stresses in the device remain safe.

A novel implantable electromechanical ventricular assist device. First acute animal testing.

A novel ventricular assist device (HIA-EMLVAD-AT1, Helmholtz Institute Aachen-Electromechanical Left Ventricular Assist Device-Animal Test Version 1), driven by a uniformly and unidirectionally rotating actuator and a patented hypocycloidic pusherplate displacement gear unit, was developed and tested in an acute animal experiment. The excellent free filling behavior of the pump chamber with a stroke volume of 65 ml is obtained by a 2:3 ejection-filling time relationship. The uniform motor rotation facilitates simple sensorless pre and afterload detection by motor current analysis. In contrast to common apical cannulation, the inlet cannula was placed via the left atrium through the mitral valve into the left ventricle. This connection mode is preferable for left ventricular recovery and preservation. Left atrial, left ventricular, and aortic pressure curves, as well as pulmonary artery flow data, were obtained. The data show very effective unloading of the natural ventricle and demonstrate the feasibility of this novel assist device. Directions for further improvement of technical features were also identified.

Precision, comfort and mechanical performance of the Quadriso-tester, a quadriceps force measuring device.

The mechanical performance, repeatability and comfort of the Quadriso-tester, which assesses isometric knee extensor muscle strength, were examined. Twenty healthy subjects and 20 patients treated for unilateral anterior ligamenta cruciata insufficiency were tested. Intra-rater repeatability was determined by the testing and retesting of subjects and calculation of the intra-class correlation coefficient and the mean difference between test and retest values. The comfort level was determined a questionnaire. Measuring time was recorded, and the relationship between knee angle and extension moment was plotted. Strength and stiffness were determined using the finite element method. Intra-rater repeatability was high; the intra-class correlation coefficient of the right and left leg was 0.90 and 0.91, respectively; the coefficient of variation was 6.4 and 6.0%, respectively. The median comfort score of the healthy subjects was 7, and that of the patients was 9. Measuring time remained within 30 min. Misalignment of the knee and sidebar axis disturbed the relationship between knee angle and extension moment Strength and stiffness were higher than required. In conclusion, the Quadriso-tester is a comfortable and fast device to determine quadriceps force with a high repeatability. The knee and sidebar axis should be well aligned.

Medical technology assessment: the use of the analytic hierarchy process as a tool for multidisciplinary evaluation of medical devices.

Most types of medical technology assessment are performed only after the technology has been developed. Consequently, they have only minor effects on changes in clinical practice. Our study introduces a new method of constructive medical technology assessment that can change the development and diffusion of a medical device to improve its later clinical effectiveness. The method, based on Saaty's Analytic Hierarchy Process, quantitatively supports discussions between various parties involved in technological development and diffusion. We applied this method in comparing a new blood pump with two competitors based on technical, medical and social requirements. These discussions changed the evaluators' perspectives, reduced diasagreements, and ended in a reliable evaluation of the pump's performance. On the basis of these results, adaptations were derived which improved the design and diffusion of the blood pump. This application shows the adequate potential of our method to steer technological development and diffusion of artificial organs.