RESUMO
Colonization of in-dwelling catheters by microbial biofilms is a major concern in patient health eventually leading to catheter-related blood stream infections. Biofilms are less susceptible to standard antibiotic therapies that are effective against planktonic bacteria. Standard procedure for the detection of microorganisms on the catheter tip is culture. However, viable but non-culturable cells (VBNCs) may be missed. The aim of this study was to evaluate the use of fluorescence in situ hybridization (FISH) as an indicator to visualize and quantify the effect of the antibiotics daptomycin and vancomycin on biofilms in situ. We established an in vitro catheter biofilm model of Staphylococcus epidermidis biofilms on polyurethane catheters. Biofilm activity was measured by FISH and correlated to colony forming units (CFU) data. Digital image analysis was used for quantification of total biofilm mass and the area of the FISH positive biofilm cells. FISH showed a pronounced effect of both antibiotics on the biofilms, with daptomycin having a significantly stronger effect in terms of both reduction of biofilm mass and number of FISH-positive cells. This supports the anti-biofilm capacity of daptomycin. Interestingly, neither antibiotic was able to eradicate all of the FISH-positive cells. In summary, FISH succeeded in visualization, quantification, and localization of antibiotic activity on biofilms. This technique adds a new tool to the arsenal of test systems for anti-biofilm compounds. FISH is a valuable complementary technique to CFU since it can be highly standardized and provides information on biofilm architecture and quantity and localization of survivor cells.
Assuntos
Biofilmes/efeitos dos fármacos , Daptomicina/farmacologia , Hibridização in Situ Fluorescente , Staphylococcus epidermidis/efeitos dos fármacos , Staphylococcus epidermidis/fisiologia , Vancomicina/farmacologia , Antibacterianos/farmacologia , Reatores Biológicos/microbiologia , Cateteres de Demora/microbiologia , Contagem de Colônia Microbiana , Processamento de Imagem Assistida por Computador , Staphylococcus epidermidis/crescimento & desenvolvimentoRESUMO
The assessment of flow along a vaulted wall (with two main finite radii of curvature) is of general interest; in biofluid mechanics, it is of special interest. Unlike the geometry of flows in engineering, flow geometry in nature is often determined by vaulted walls. Specifically the flow adjacent to the wall of blood vessels is particularly interesting since this is where either thrombi are formed or atherosclerosis develops. Current measurement methods have problems assessing the flow along vaulted walls. In contrast with conventional particle image velocimetry (PIV), this new method, called wall PIV, allows the investigation of a flow adjacent to transparent flexible surfaces with two finite radii of curvature. Using an optical method which allows the observation of particles up to a predefined depth enables the visualization solely of the boundary layer flow. This is accomplished by adding a specific dye to the fluid which absorbs the monochromatic light used to illuminate the region of observation. The obtained images can be analysed with the methods of conventional PIV and result in a vector field of the velocities along the wall. With wall PIV, the steady flow adjacent to the vaulted wall of a blood pump was investigated and the resulting velocity field as well as the velocity fluctuations were assessed.
Assuntos
Artérias/fisiologia , Velocidade do Fluxo Sanguíneo/fisiologia , Meios de Contraste , Interpretação de Imagem Assistida por Computador/métodos , Microscopia de Fluorescência/métodos , Reologia/métodos , Tamanho da PartículaRESUMO
Insemination with the sperm of an epididymal reservoir three days after implantation of the prosthesis led to a successful pregnancy with subsequent delivery of five healthy sucklings. Alloplastic spermatocele can function longer than four months as proved by a successful pregnancy.
Assuntos
Epididimo , Inseminação Artificial , Próteses e Implantes , Espermatozoides , Animais , Feminino , Inseminação Artificial/métodos , Masculino , Gravidez , SuínosRESUMO
Atherosclerosis related cardiovascular diseases are the leading cause of death in western societies. The clinical manifestations are chronic arterial obstructions or acute arterial occlusions in various vascular territories. The pathogenesis is only understood in part as yet. Arterial wall abnormalities, blood composition abnormalities and hemodynamic alterations are generally accepted to be causative (Virchow's triad). The key role is played by macrophages in the subendothelial space that are activated immunologically by oxidized LDL particles via the scavenger receptor pathway. Recently, endothelial dysfunction due to oxidative stress was identified as a priming factor in the course of the development of atherosclerotic plaques. Shear stress-induced microinjuries of the endothelium in hemodynamically compromised regions together with local coagulation activation associated with microinflammation of the plaque are currently thought to cause plaque rupture. This event is the reason for local clot formation and ultimate organ infarction. Treatment success is still insufficient, however some progress during the last decade is reflected by the improving outcome of atherosclerosis associated cardiovascular diseases. Evidence from clinical trials supports the efficacy of statins, antiplatelet agents, antihypertensive agents if necessary and omega-fatty acids in patients with overt atherosclerosis. The reduction of mortality achieved by those drugs amounts to: omega-fatty acids -21%, statins -16%, anti-platelet agents -14%, treatment of hypertension -13%. It is impossible to calculate the combined effect of these modalities since in each trial participants received co-medication containing agents tested in other trials.
Assuntos
Arteriosclerose/tratamento farmacológico , Arteriosclerose/fisiopatologia , Ensaios Clínicos como Assunto , Feminino , Humanos , Masculino , Resultado do TratamentoRESUMO
The carotid artery bifurcation is of special interest to both the pathologist because of its frequent atherosclerotic depositions, and to the fluidmechanicist because of its complex flow properties. However, current knowledge is incomplete regarding the level and position of atherosclerotic plaques in the carotid bifurcation and their quantitative correlation with its geometry. The study presented here is intended to fill that knowledge gap and try to quantify the geometrical risk factors. During the post-mortem the arteries were excised and filled with a resin at the proper transmural pressure of 80 mm Hg. Thirty-one vessel casts of the carotid artery from twenty-three autopsied individuals were made. The vessel casts were used to measure several geometrical parameters. After fabrication of the vessel casts each artery was investigated according to pathomorphological procedure. An atherometric system (AS) indicating the level of atherosclerotic lesions was applied. The specimens were divided into three groups according to the level of atherosclerosis. The comparison was made between the level of wall alteration of the main branches of the carotid bifurcation (common, internal and external carotid branch), and between these three groups themselves. Further, we conducted a comparison of the averaged geometric parameters in different groups to define the correlations between atherosclerotic lesions and geometric parameters. The results show that the most advanced lesions (fibrous and severe plaques) with about 70% of all lesions were mainly found in the internal and the external carotid branches, compared with only 25% for the common carotid branch. The comparative analysis showed that a relatively high carotid sinus enlargement of > 1.2 of the common carotid branch diameter is a most significant geometric risk factor among those investigated, whereas there was no correlation between branch angles and atherosclerosis. In conclusion, the quantification of atherosclerotic risk factors is very important in the investigation of atherosclerotic disease development.
Assuntos
Arteriosclerose/patologia , Doenças das Artérias Carótidas/patologia , Artéria Carótida Primitiva/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Arteriosclerose/classificação , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
The carotid artery is of special interest for the pathologist because of its frequent depositions, and for the fluidmechanician because of its complex flow properties. However, there is a distinct lack in current knowledge of its geometry. Therefore, a vessel cast study was undertaken. At post mortem, the arteries are excised and filled with a special resin at the proper transmural pressure. Eighty-six vessel casts of the carotid artery were performed, and some etiological factors of atherosclerosis, such as age, sex and disease, were collected. The following selected geometric parameters of these vessel casts were measured in this study: the diameters of the main branches of carotid bifurcation (common, internal and external arteries), and the angles between internal, external and common carotid arteries. The averaged geometric parameters and their variability over 86 vessel casts of the carotid artery were investigated. Furthermore, the relationship between these measured parameters and the etiological factors age, sex and disease was analyzed. The geometric parameters varied considerably, presumably contributing to a corresponding variability in the local hemodynamic and distribution of the atherosclerotic lesions.
Assuntos
Artéria Carótida Primitiva/anatomia & histologia , Idoso , Idoso de 80 Anos ou mais , Doenças das Artérias Carótidas/patologia , Artéria Carótida Primitiva/patologia , Molde por Corrosão , Feminino , Humanos , Hipertensão/patologia , Masculino , Matemática , Pessoa de Meia-IdadeRESUMO
The carotid artery is of special interest both for the pathologist because of its frequent depositions and for the fluid mechanic because of its complex flow properties. However, current knowledge of its geometry is insufficient. Therefore, a vessel cast study was undertaken and a method to fabricate vessel casts was developed. At post mortem the arteries are excised and filled with a resin at the proper transmural pressure. Thirty-one vessel casts of the carotid artery were performed and the following selected geometric parameters of these vessel casts were measured: the diameters of the main branches of carotid bifurcation (common, internal and external arteries) and the angles between internal, external and common carotid arteries. The geometric parameters vary considerably and may contribute to a corresponding variability in local hemodynamics.
Assuntos
Artéria Carótida Primitiva/patologia , Molde por Corrosão , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fatores de RiscoRESUMO
Clinical applications of cardiac assist systems continue to have a severe problem, that of thromboembolic complications. The problem originates mainly at the valves, which are usually made of a antithrombogenic material, such as bovine pericardium. However, the valve housing is made of a less suitable material, and wherever the blood flow is stagnant, a thrombus is likely to form. Such stagnant blood flow is found in the space between the housing of the valve and the leaflets, in the sinuses. Consequently, thrombi often are generated in the sinuses. The novel valve design presented in this article avoids the formation of the stagnation zone in the sinuses by a purge flow. This flow is taken from the main flow through the valve and is directed into each sinus of the res purges the sinuses. The purge flow effect is investigated with an experimental method in which the sinus is filled with dye, and washout during the valve action is observed and recorded on videotape, which is compared with washout in a valve without a purge flow. In addition, the purge flow effect is investigated by computational fluid dynamics. Both methods show that the purge flow effectively increases fluid exchange in the sinuses.
Assuntos
Próteses Valvulares Cardíacas , Simulação por Computador , Desenho de Prótese , Fluxo Sanguíneo RegionalRESUMO
The blood flow immediately adjacent to the wall of a blood vessel or an artificial surface is of great interest. This flow defines the shear stress at the wall and is known to have a great physiological importance. The use of models is a viable method to investigate this flow. However, even in models the shear stress at the wall is difficult to assess. A new optical method is based on transparent models and uses particles in the model fluid, which are only visible near the wall. This is achieved with a model fluid having a defined opacity. This fluid obscures particles in the center of the models, but permits the observation and recording of particles close to the wall. The method has been applied for Hagen-Poiseuille flow and for the likewise well researched flow in a tube with a sudden expansion.
Assuntos
Artérias/fisiopatologia , Endotélio Vascular/fisiopatologia , Modelos Cardiovasculares , Hemorreologia , Humanos , Fluxo Sanguíneo Regional , Estresse MecânicoRESUMO
The influence of blood flow on the depositions and development of atherosclerotic lesions have been observed and described since the 19th century. Observations have shown that depositions correlate with regions of low wall shear stress. However, the exact correlations between depositions, vessel geometry and flow parameters are not yet known. The purpose of this study was the quantification of atherosclerosis risk factors in carotid bifurcation. This artery has attracted particular interest because lesions are often found in this bifurcation. Post mortem, the arteries are excised and vessel casts are produced. Afterwards, the arteries are analyzed morphometrically. The vessel casts are used for the assessment of some geometrical parameters. 31 carotid bifurcations were analyzed in this study. Eight vessel casts were digitized and rendered three-dimensional mathematical models of the arteries. These data were imported by the computational fluid dynamics program FLUENT. Further, the blood flow was reconstructed in a computer model based on the individual vessel geometry. The flow parameters, such as velocity, pressure and wall shear stress were computed. At the same time the geometrical parameters and wall alterations are known. This permits the comparison of the anatomical shape and its flow with the distribution and level of the wall alterations.
Assuntos
Artérias Carótidas/patologia , Doenças das Artérias Carótidas/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Artérias Carótidas/fisiopatologia , Doenças das Artérias Carótidas/sangue , Doenças das Artérias Carótidas/fisiopatologia , Molde por Corrosão , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fluxo Sanguíneo Regional , Estresse MecânicoRESUMO
Numerous devices and mock circulations have been described for the measurement of pressure loss, closure time, closing and leakage volumes and energy loss in artificial heart valves. However, all the devices have been troubled with difficulties in generating and assessing the precise flow through the valve, and problems in defining the arterial load, i.e. the artificial aorta. The new test device follows a radically different approach: a computer controlled piston forces the fluid through the test valve only--with no afterload. During systole, outflow follows a physiological curve which is identical for all types of heart valves of a given size. During diastole a mathematically defined physiological pressure difference curve is followed. Consequently, the measurements are independent of the individual machine, the lab where testing takes place, the scientist who executes the test, the time when measurements are taken and all other external influences.
Assuntos
Próteses Valvulares Cardíacas , Hemodinâmica , Microcomputadores , Modelos Estruturais , Desenho de Equipamento , Humanos , Técnicas In Vitro , Pulso Arterial , Controle de QualidadeRESUMO
An organ's vitality prior to transplantation can be evaluated by analysis of its mechanical properties. A new apparatus was designed to measure these properties. The organ to be tested is placed in a reservoir with saline and submitted to the load of a cylindrical element. The load is applied in a stepwise mode and results in a small local deformation of the organ. This deformation is identical to the displacement of the cylinder and is measured as a function of time. This is compared to a theoretical analysis of the deformation of a viscoelastic halfspace. This theory was used to interpret the results of experiments which were carried out on eight rabbit kidneys. The instantaneous modulus of elasticity reaches a maximum within 15 minutes, then decreases and reaches a steady state after 30 minutes of warm ischemia. The method is easy to apply and appears to be useful for the quantification of an organ's vitality and in particular recognizes the level of the organ's edema before transplantation.
Assuntos
Transplante de Rim , Animais , Fenômenos Biomecânicos , Sobrevivência Celular , Elasticidade , Coelhos , ViscosidadeRESUMO
Clinical applications of cardiac assist systems still suffer from thromboembolic complications due to thrombus formation behind the valve's leaflets. Wherever the flow is stagnant such as in the sinus, a thrombus is likely to form. The valve design in this study avoids stagnation zones behind the leaflets by a purge flow during systole. This purge flow is separated from the valve's main flow by a flow divider directing a part of the main flow into the sinus behind the leaflet. The optimization was performed on a monoleaflet valve because of its simple geometry. Relevant geometric parameters were systematically varied. Thirty-two models were designed and numerically investigated. The models with the best results were preselected and investigated in a computer-controlled valve tester. The washout of a dye filled in the sinus was digitally recorded and quantified. The results show that a sinus purge flow minimizes stagnation areas in the sinus and thus may avoid thrombus formation.
Assuntos
Circulação Coronária , Próteses Valvulares Cardíacas/normas , Desenho de Prótese/métodos , Simulação por Computador , HumanosRESUMO
Atherosclerotic ailments play a major role in industrial societies. Atherosclerotic disease causes stenotic narrowing of the arterial vessel system. These narrow passages can be widened with balloon angioplasty and stents are introduced to keep the passage open. In this study, stents designed for the aorto-iliac bifurcation are considered. Most of these stents used in peripheral arteries are self-expanding stents made from Nitinol, a thermo-reactive alloy. The insertion is done under radiographic control. However, the delicate metallic structure of the stent usually casts only a weak radiographic shadow and thus is difficult to detect. A stent with an innovative radio opaque tantalum marker (Luminexx, C.R. BARD--Angiomed, Karlsruhe), overcomes this problem. However, the markers extend into the blood flow and the question arises whether the markers may cause the generation of thrombi. In a model study in an enlarged model of the aorto-iliac bifurcation with an inserted stent the flow was visualized. The enlarged scale permits the observation and video recording of the flow in great detail. The Reynolds similarity is kept. A subsequent analysis of the flow in the vicinity of the markers shows a short residence time. These results indicate that the additional markers do not increase the risk of thrombus generation.
Assuntos
Aorta/patologia , Arteriosclerose/fisiopatologia , Arteriosclerose/terapia , Hemorreologia , Artéria Ilíaca/patologia , Stents , Ligas/efeitos adversos , Velocidade do Fluxo Sanguíneo , Humanos , Processamento de Imagem Assistida por Computador , Modelos Biológicos , Modelos Cardiovasculares , Trombose/etiologiaRESUMO
The flow curve of a valve-tester has a great influence on the performance of a valve. Usually the flow curve of the aorta is used. However, the mitral valve flow curve differs greatly from the flow curve of the aortic valve. It varies with the pulse rate and is further changed in patients with certain heart diseases. To investigate the different mitral flow conditions, ultrasonic flow curves from patients with a mechanical artificial mitral valve were analyzed. The curves show that a mitral valve prosthesis has not only to work under physiological flow conditions, but also in pathologically deviant flows. According to these results three different characteristic flow curves were selected and used to test several valves with a computer controlled valve-tester. The mean diastolic pressure difference and the whole closing behavior were influenced by the flow curve; and the differences in energy losses were particularly great. This indicates, that the flow curve must be adjusted appropriately.
Assuntos
Próteses Valvulares Cardíacas/normas , Modelos Cardiovasculares , Aorta , Valva Aórtica , Fenômenos Biomecânicos , Ventrículos do Coração , Humanos , Valva Mitral , Volume Sistólico , UltrassomRESUMO
An investigation was performed to demonstrate that time-lapse cinematography and computer aided video analysis of cell morphology is suitable to study and compare the characteristics of hepatocytes during the adhesion process to membranes. We chose to compare ordinary cellulose Cuprophan membranes and membranes coated with collagen or fibronectin. Striking differences between uncoated cellulose and fibronectin or collagen coating were seen in the cell count per square millimeter and adhesion behaviour. On the investigated uncoated Cuprophan the hepatocytes were found to attach but not to spread whilst on collagen coated Cuprophan most of the cells spread spherically, and on fibronectin coated membranes most of the cells flattened spherically or polygonally. Time-lapse video microscopy seems to be a valuable technique for assessing the morphologic behaviour of cells in a detailed and quantitative manner in order to improve the hepatocyte culture technique in bioreactors for hybrid systems.
Assuntos
Fígado/citologia , Membranas Artificiais , Animais , Materiais Biocompatíveis , Adesão Celular , Celulose/análogos & derivados , Colágeno , Fibronectinas , Processamento de Imagem Assistida por Computador , Masculino , Ratos , Ratos Endogâmicos , Gravação de VideoteipeRESUMO
Coronary artery bypass graft (CABG) operation for coronary artery disease with different types of grafts has a large clinical application world wide. Immediately after this operation patients are usually relieved of their chest pain and have improved cardiac function. However, after a while, these bypass grafts may fail due to for example, neointimal hyperplasia or thrombosis. One of the causes for this bypass graft failure is assumed to be the blood flow with low wall shear stress. The aim of this research is to estimate the wall shear stress in a graft and thus to locate areas were wall shear stress is low. This was done with the help of a blood flow computer model. Post-operative biplane angiograms of the graft were recorded, and from these the three-dimensional geometry of the graft was reconstructed and imported into the computational fluid dynamics (CFD) program FLUENT. The stationary diastolic flow through the grafts was calculated, and the wall shear stress distribution was estimated. This procedure was carried out for one native vessel and two different types of bypass grafts. One bypass graft was a saphenous vein and the other one was a varicose saphenous vein encased in a fine, flexible metal mesh. The mesh was attached to give the graft a defined diameter. The computational results show that each graft has distinct areas of low wall shear stress. The graft with the metal mesh has an area of low wall shear stress (< 1 Pa, stationary flow), which is four times smaller than the respective areas in the other graft and in the native vessel. This is thought to be caused by the smaller and more uniform diameter of the metal mesh-reinforced graft.
Assuntos
Simulação por Computador , Ponte de Artéria Coronária , Doença das Coronárias/cirurgia , Hemorreologia , Modelos Cardiovasculares , Angiografia Coronária , Diástole/fisiologia , Humanos , Hiperplasia/prevenção & controle , Imageamento Tridimensional , Estresse Mecânico , Telas Cirúrgicas , Trombose/prevenção & controle , Túnica Íntima/patologia , Grau de Desobstrução VascularRESUMO
Platelet deposition in resting blood is well researched and understood. However, the influence of hemodynamic parameters such as wall shear rate is less clear. Clinical experience and experiments show an interaction between flow and platelet deposition. But a complete understanding of the flow influence and hence a quantification has not yet been achieved. A well defined experiment of flow dependant platelet depositions is the stagnation point flow. This kind of flow is ubiquitous in the circulatory system, to be found in every bifurcation and recirculation region. These are the areas where thrombus formations are likely to occur if other conditions are also met. First, experiments were performed in a stagnation point flow chamber. A simplified blood model, platelet rich plasma, was used as a test fluid. With a microscope the platelet deposition was observed and recorded. Platelets deposit in a characteristic pattern showing the influence of the flow. An analysis of this pattern is the objective of this study and is achieved with the help of a numerical model, which is based on a convective diffusive transport. The model results in a platelet deposition pattern, which in its shape and temporal development is very similar to experimental results. Hence it is concluded that the assumed transport processes are causal for platelet depositions and thrombus formation.
Assuntos
Simulação por Computador , Hemorreologia , Modelos Cardiovasculares , Adesividade Plaquetária/fisiologia , Animais , Cães , Método de Monte Carlo , Trombose/metabolismoRESUMO
Even nowadays, the essential problem of mechanical heart valve prostheses is the risk of thromboembolic events mainly caused by unnatural hemodynamics, e.g. just a few years ago the Medtronic Parallel (MP) showed unsatisfactory clinical results caused by thrombi. Therefore, in vitro investigations of the whole leakage jets were performed at the MP in mitral position by means of a pulse duplicator using a two channel laser Doppler anemometer. From the measured data, mean velocity profiles and the distribution of Reynolds shear stresses, as a function of the location within the jet, were calculated. From this data the potential of blood damage is evaluated computing a Blood Damage Index (BDI) of hemolysis and platelet damage. Four regurgitant free jets right above the hinges were observed during systole at the inflow side of the MP. The peak velocities at the origin of the jets were in the order of 1.6-2.1 m/s. Two jets experienced maximum turbulent shear stresses around 100 N/m2 within this area. The BDI for platelets of the MP is around ten times higher than the BDI of the St.-Jude-Medical. The study shows that besides the flow structure within the hinges of a mechanical heart valve, the whole regurgitant jet has a large blood damage potential. This potential is measurable, respectively calculable and seems to be (on account of it's support of the clinical outcome) one piece of the puzzle that explains the negative trials of the MP.
Assuntos
Próteses Valvulares Cardíacas/efeitos adversos , Velocidade do Fluxo Sanguíneo , Plaquetas , Análise de Falha de Equipamento , Hemodinâmica/fisiologia , Hemólise , Humanos , Técnicas In Vitro , Fluxometria por Laser-Doppler , Desenho de Prótese , Falha de Prótese , Estresse MecânicoRESUMO
BACKGROUND: Hydrodynamic performance of a decellularized pulmonary porcine valve was evaluated with a computer versatile pulse duplicator and compared to glutaraldehyde fixated stentless porcine bioprosthesis and a polyurethane heart valve. METHODS: Decellularized pulmonary porcine matrices (Group I, n = 5) were treated chemically to become cell-free collagen matrices. The findings of this heart valve were compared with aortic glutaraldehyde treated porcine prostheses (Group II, n = 5) and polyurethane three leaflet valve prostheses (Group III, n = 1). Measurements were performed in 0.9% saline test fluid at room temperature. Measurements compared were closing time, closing volume, systemic pressure difference and energy losses. Each valve was measured 6 times with 70 beats/minute, a stroke volume of 70 ml corresponds to a cardiac output of 4.9 L/minute. RESULTS: Group I and group III showed no significant differences between parameters. The measured closing time was significantly different (p < 0.001) between group I and II, respectively 24.333 and 53.600 ms and group II and III respectively 53.600 and 24.000. Difference in closing volume was significant (p < 0.05) between groups II and I respectively 3.67 and 0.68 ms and group II and III respectively 3.67 and 0.71. Systolic mean pressure gradient was 18.25 +/- 1.04 mm Hg in group II which was significantly different (p < 0.001) from groups I and III, respectively 10.65 +/- 0.29 mm Hg and 7.70 +/- 0.30 mm Hg. CONCLUSIONS: Decellularized pulmonary porcine valves showed the same excellent performance as polyurethane valve prosthesis, which are superior to the investigated glutaraldehyde fixed xenograft.