Micron scale directional coupler as a transducer for biochemical sensing.

A microfabricated directional coupler (DC) was used for the detection of DNA conjugated with quantum dots. Output optical signals from DCs of a wide range of device lengths correspond well to theoretical and simulation results. Even 20 µm-long DC devices could detect changes in the output optical intensity by monitoring the near-field pattern using a CCD camera. The signal was enhanced 60 × using a 1500 µm-long DC device. For large cladding refractive-index changes between air and water, the normalized signal changed cyclically several times between 0 and 1. The results suggest that the DC can be the basis for miniaturized two-dimensionally integrated biochemical sensors.

Radiation-induced xerostomia: objective evaluation of salivary gland injury using MR sialography.

BACKGROUND AND PURPOSE: Xerostomia (dry mouth) is one of the serious complications of head and neck irradiation and has a strong influence on a patient's activities of daily living. MR sialography with salivary secretion stimulation provides additional functional information (salivary secretion reserve) and may contribute to the evaluation of the severity of xerostomia and predict the risk of developing a radiation-induced xerostomia. This aim of the study was to analyze MR sialography as an objective tool to evaluate radiation-induced salivary injury. MATERIALS AND METHODS: MR sialography with salivary secretion stimulation was performed in 16 patients with head and neck malignancy before and after irradiation therapy. Multivariate (stepwise multiple regression) analysis was performed to analyze the nonstimulated and stimulated MR sialography findings and the clinical severity of xerostomia. RESULTS: Multivariate analysis of the preirradiation study revealed no significant independent variables that could predict the clinical severity of xerostomia. In the postirradiation study, following regression with 2 independent variables (secretion response of the submandibular gland [rSG] and parotid gland visualization on stimulated MR sialography [sPG]) could explain 70% of the cases: xerostomia severity grade = 0.681 + 0.871 x rSG - 0.471 x sPG. CONCLUSIONS: MR sialography is a useful method for visualization of salivary gland radiation injury and estimation of the severity of radiation-induced xerostomia. Insufficiency of secretion reserve at the irradiated submandibular gland has the strongest influence on xerostomia severity. Our investigation suggests that careful submandibular gland protection may lead to prevention and avoidance of radiation-induced xerostomia.

Mechanical properties of plasma membrane and nuclear envelope measured by scanning probe microscope.

Atomic force microscopy has been used to visualize nano-scale structures of various cellular components and to characterize mechanical properties of biomolecules. In spite of its ability to measure non-fixed samples in liquid, the application of AFM for living cell manipulation has been hampered by the lack of knowledge of the mechanical properties of living cells. In this study, we successfully combine AFM imaging and force measurement to characterize the mechanical properties of the plasma membrane and the nuclear envelope of living HeLa cells in a culture medium. We examine cantilevers with different physical properties (spring constant, tip angle and length) to find out the one suitable for living cell imaging and manipulation. Our results of elasticity measurement revealed that both the plasma membrane and the nuclear envelope are soft enough to absorb a large deformation by the AFM probe. The penetrations of the plasma membrane and the nuclear envelope were possible when the probe indents the cell membranes far down close to a hard glass surface. These results provide useful information to the development of single-cell manipulation techniques.

Fast-scanning atomic force microscopy reveals the molecular mechanism of DNA cleavage by ApaI endonuclease.

Newly developed fast-scanning atomic force microscopy (AFM) allows the dissection of molecular events such as DNA-enzyme reactions at the single-molecule level. With this novel technology, a model is proposed of the DNA cleavage reaction by a type IIP restriction endonuclease ApaI. Detailed analyses revealed that ApaI bound to DNA as a dimer and slid along DNA in a one-dimensional diffusion manner. When it encountered a specific DNA sequence, the enzyme halted for a moment to digest the DNA. Immediately after digestion, the ApaI dimer separated into two monomers, each of which remained on the DNA end and then dissociated from the DNA end. Thus, fast-scanning AFM is a powerful tool to aid the understanding of protein structures and dynamics in biological reactions at the single-molecule level in sub-seconds.

Identifying patterns of spatial current dispersion that characterise and separate the Brugada syndrome and complete right-bundle branch block.

The aim of the study was to detect patterns of spatial-current distribution in the late QRS and early ST-segments that distinguish Brugada-syndrome cases from complete right-bundle branch block (CRBBB). Magnetocardiograms (MCGs) were recorded from Brugada-syndrome patients (n = 6), CRBBB patients (n = 4) and the members of a control group (n = 33). The current distributions at six time points from Q-onset were estimated by producing current-arrow maps (CAMs). The angle of the current arrow of maximum amplitude at each time point was calculated. In the Brugada cases, the characteristic ST elevation was seen above the upper right chest, and abnormal currents appeared to be present in the right-ventricular outflow tract (RVOT). The angles of the abnormal arrows were -78 degrees +/- 51 degrees at 100 ms and -50 degrees +/- 61 degrees at 110 ms. In the cases of CRBBB, wide S- and R-waves were recorded above the upper right and lower right chest, respectively. The angles of the abnormal arrows for CRBBB were 152 degrees +/- 19 degrees at 100 ms, 159 degrees +/- 20 degrees at 110 ms, and 157 degrees +/- 19 degrees at 120 ms. The findings suggest that an abnormal current from the RVOT to the upper left chest may be a feature of the Brugada syndrome, and that the direction of this current is completely different from that seen in CRBBB.

Morphologic changes in the aorta during elastase infusion in the rat aneurysm model.

BACKGROUND: The morphologic changes in the aorta during elastase infusion, which have not previously been investigated, were examined in the rat abdominal aortic aneurysm model. MATERIALS AND METHODS: A total of 80 Wistar rats were divided into five groups. A 1.0-cm segment of infra-renal abdominal aorta was infused with 25 U of elastase in 1 mL saline for 10 (n = 14), 20 (n = 14), 30 (n = 14), 60 (n = 19), or 120 min (n = 19). In the 120-min group, transparency and aortic diameter were recorded every 10 min. In each group, 7 rats were killed immediately after infusion. The aortas were excised for histologic examination. The aortic diameters were measured 7 days after infusion in the remaining rats. RESULTS: The infused aorta became transparent within 50 min of elastase infusion. The aortic diameter increased rapidly for the first 30 min of infusion. Histologically, the elastic tissue was completely absent after 60 min of infusion. The aortic diameters in the 60- and 120-min groups were not significantly different 7 days after infusion. CONCLUSIONS: Morphologic changes in the infused aorta are complete after 60 min of elastase infusion. It may be possible to shorten the elastase infusion time from 120 to 60 min in this model.

Factors influencing mortality in the rat elastase-induced-aneurysm model.

BACKGROUND: Intraarterial infusion of elastase has been used to create an experimental model of aortic aneurysm in rats. Unfortunately, the utility of this model is limited by a high mortality rate among experimental animals. This study examined the factors influencing mortality to help refine this model. MATERIALS AND METHODS: A total of 126 Wistar rats were divided into six groups. A 1.0-cm segment of infra-renal abdominal aorta was clamped (n = 21), clamped and cannulated via the femoral artery (n = 21), infused with saline (n = 21), or infused with solution containing 25 U of elastase from three different lots (each group, n = 21). Clamping or infusion was performed for 30 (n = 7), 60 (n = 7), and 120 min (n = 7). The mortality rates were calculated at 7 days. RESULTS: There were no deaths in the clamp group, but 29% of the rats that underwent cannulation with clamping or saline infusion for 2 h died. The mortality rate for a 30-min elastase infusion ranged from 0 to 71%. Mortality for a 60-min infusion ranged from 14 to 100%. Mortality for a 2-h infusion ranged from 43 to 100%. The mortality rate was dependent on the treatment time and the elastase lot number. CONCLUSIONS: Aortic cannulation, elastase infusion, and prolonged infusion times all increase the mortality rate in the elastase-induced rat aortic aneurysm model. Mortality is also dependent on the elastase lot number.

Impeller design for a miniaturized centrifugal blood pump.

The impeller design for a miniature centrifugal blood pump is an important consideration since the small diameter impeller requires higher rotational speed, which may cause more blood trauma compared to the larger diameter impeller. Three different impeller vanes (straight vanes with a height of 4 mm and 8 mm, and 8 mm curved vanes) of which the diameter was 35 mm were subjected to hydraulic performance and hemolysis tests in the same pump housing. Both straight vane impellers attained left ventricular assist condition (5 L/min against 100 mm Hg) at 2,900 rpm while the curved vane required 3,280 rpm. There was no significant hemolysis difference between the tall and short vanes. The curved impeller vanes did not exhibit sufficient hydraulic performance when compared to the straight vanes. The straight vane impellers, even with different heights, were incorporated into the same pump housings, and the vane heights did not drastically change the hydraulic performance or hemolysis.

Pharmacokinetics of a parenteral carbapenem, biapenem, in patients with end-stage renal disease and influence of haemodialysis.

The effects of haemodialysis on the pharmacokinetics of a carbapenem, biapenem, were evaluated in five patients with end-stage renal disease, who received 1 h iv infusions of 300 mg biapenem on both the days on and off 4 h haemodialysis. With haemodialysis, plasma biapenem exhibited two elimination phases, one during and the other after haemodialysis with half-lives of 1.16 +/- 0.12 and 3.33 +/- 0. 91 h, respectively. Ninety percent of biapenem was removed from blood to dialysate. Without haemodialysis, plasma biapenem was mono-exponentially eliminated with a half-life of 4.35 +/- 1.30 h.

Control system for an implantable rotary blood pump.

Rotary blood pumps can be used for long-term left ventricular assist devices. These pumps have several advantages over the conventional pulsatile pumps including smaller size, higher efficiency, and simple design and construction. However, one of the difficulties associated with the rotary blood pump is the proper control method to maintain an optimum flow rate in different physiological conditions. The rotary blood pump can be controlled by two methods. The first is to utilize the measured pump flow rate from its servo signal. The second is to detect and avoid abnormal pumping conditions such as; back flow and sudden increase in the pressure head. This abnormal situation typically occurs from excessive suction of blood when there is a functional or mechanical occlusion in the inflow cannula. The ultrasound flow meter is durable and reliable but it is difficult to continually monitor the blood flow rate of an implantable pump. Therefore, another method is needed instead of the continuous flow monitoring. One chronic calf having an LVAD was subjected for the development of this control system. This calf survived more than 6 months. Voltage, current, motor speed, heart rate and the pump flow rate were recorded and stored at 30-min intervals in a computer. Utilizing these parameters, attempts were made (1) to achieve indirect flow assessments and (2) to reveal abnormal operating parameters of the centrifugal pump (1). Indirect flow measurement, the predicted pump flow rate was calculated from these pump derived parameters (required power, motor speed and heart rate). The value of the coefficient of determination (R) between the measured and estimated pump flow rate was 0.796. (2) Abnormal operating indicator, there was an association between the required current and pump flow waves. The current was differentiated, and then calculated to the power of the differentiated current. The normal range of this value was 0.02+/-0.54. In abnormal conditions, this abnormal operating indicator increased 500 times. The predicted flow estimation method and abnormal operating indicator were available from intrinsic operating parameters of the pump and need no sensors. These two methods were simple, yet they are possibly effective and reliable servo control methods for a rotary blood pump.

Gyro pump wear and deformation analysis in vivo study: creep deformation.

The Gyro pump has a double pivot bearing system to support its impeller. In this study, the integrity of the bearing system was examined after ex vivo studies. The pumps were implanted into calves and evaluated for different periods as a paracorporeal left ventricular assist device (LVAD). One pump was subjected to a test of 30 days, 1 for 15 days, 4 for 14 days, 1 for 10 days, 1 for 7 days, 2 for 4 days, and 4 for 2 days. One additional pump was subjected to percutaneous cardiopulmonary support (PCPS) condition for 6 days (total pressure head 500 mm Hg with a pump flow rate of 3 L/min). The anticoagulation treatment consisted of a continuous administration of heparin to maintain an achieved clotting time (ACT) of 200-250 s during the LVAD study and 250-300 s during the PCPS study. After the experiment, the pumps were disassembled, and the wear and deformation of male and female bearings were analyzed. There were no dimensional changes on male bearings but there were on female bearings. Wear and deformation of the female bearings were calculated as follows: wear and deformation = (depth of female before pumping) - (depth after pumping). Thirteen assembled Gyro pumps were disassembled to measure the depth of the female bearings before pumping. There was no statistical relationship between the wear and deformation and the motor speed x driving period. From these results, the deformation was not due to wear but to the creep or elastic deformation. This study suggested that the double pivot bearing system of the Gyro pump is highly durable.

Antithrombogenicity evaluation of a centrifugal blood pump.

The Gyro C1E3 pump was developed not only for cardiopulmonary bypass but also as a short-term assist device. The main purpose of this study was to examine the correlation between the thrombus formation factor and the Gyro C1E3 pumps. Seven pumps were implanted into 3 calves and evaluated for different periods of duration as a paracorporeal left ventricular assist device (LVAD). One pump was subjected to percutaneous cardiopulmonary support condition (PCPS) (total pressure head 500 mm Hg with a pump flow rate of 3 L/min). The anticoagulation treatment consisted of a continuous administration of heparin to maintain an activated clotting time (ACT) of 200-250 during the LVAD study and 250-300 during the PCPS study. After the experiment, the pumps were disassembled and examined. In cases where there were any blood-derived deposits inside the pumps, the dry weight of these thrombi that adhered to the bearing area of the pump was measured. A multiple correlation was attempted to speculate possible thrombus formation. The estimated dry weight of thrombi was calculated from pump flow rate, pumping day, motor speed, and activated clotting time. This equation was estimated dry weight of thrombi = 1.140 x pump flow rate -0.001 motor speed + 1.652 pumping time -0.041 x ACT + 2.198 R2 = 0.944. This study suggested that there was a possibility to calculate the amount of adhered thrombus formation from pump flow rate, motor speed, pumping day, and ACT.

The time course of elastin fiber degeneration in a rat aneurysm model.

Previous findings vary regarding the timing and cause of elastin fiber degeneration in the elastase-induced rat abdominal aortic aneurysm model. We examined the timing and cause of elastin fiber degeneration after elastase infusion using two different elastase infusion times. Twenty-four Sprague-Dawley rats were divided into two groups. The infrarenal abdominal aorta was infused with 15 U of elastase for 15 min (n = 12, 15-min infusion group) or 30 min (n = 12, 30-min infusion group). In each group, three rats were killed immediately and 1, 3, and 7 days after infusion, and then the aortas were excised for a histologic examination. Elastin fibers did not stain, even immediately after elastase infusion, in the 30-min infusion group. The degeneration of elastin fibers did not progress in the 15-min infusion group during the period of observation. Inflammatory cells infiltrated mainly to the adventitia near regions where the degeneration of elastin fibers spread totally through the aortic media. Elastin fibers degenerate immediately after elastase infusion and thus seem to degenerate not due to endogenous proteinases that are produced by the infiltrating cells, but due to the exogenously infused elastase itself. Inflammatory cell infiltration was thus found to be a result of the degeneration of elastin fibers in this model.

Particles released from the Gyro C1E3 during simulated extracorporeal circulation.

Evaluation of released particles from the blood pump during extracorporeal circulation is an important aspect because the particles may cause microembolism. The Gyro C1E3 is a centrifugal blood pump that has an impeller suspended by double pivot bearings inside the housing; therefore, it is important to evaluate the released particles. The C1E3 was driven for 14 days to simulate clinical left ventricular assist device (LVAD) and percutaneous cardiopulmonary support (PCPS). Also, a roller pump was driven for 2 days as a comparison. Released particles were weighed and examined by SEM. After 14 days of pumping, the particles from the C1E3 were 238.6 microg in an LVAD condition. The particles with the roller pump were 270.2 microg after only 2 days. Average particle sizes with the roller pump and C1E3 were 3.7 and 0.6 microm, respectively. These results suggest that the Gyro C1E3 substantially reduces the risk of microembolism from released particles.

Estimation of pump flow rate and abnormal condition of implantable rotary blood pumps during long-term in vivo study.

The control system for an implantable rotary blood pump is not clearly defined. A detection system is considered to be necessary for pump flow monitoring and abnormal conditions such as back flow or a sucking phenomenon where the septum or left ventricle wall is sucked into the cannula, etc. The ultrasound flowmeter is durable and reliable but the control system should not be totally dependent on the flowmeter. If the flowmeter breaks, the rotary blood pumps have no control mechanism. Therefore, the authors suggest controlling the pumps by an intrinsic parameter. One left ventricular assist device (LVAD) calf model was studied where the flow rate and waveform of the pump flow proved to identify the sucking phenomenon. Thus, the pump flow rate was calculated from the required power, motor speed, and heart rate. The value of the coefficient of determination (R2) between the measured and estimated pump flow rate was 0.796. To estimate this abnormal phenomenon, 2 methods were evaluated. One method was the total pressure head in which the pump flow rate and motor speed were estimated. During normal conditions the total pressure head is 79.5 +/- 7.0 mm Hg whereas in the abnormal condition, it is 180.0 +/- 2.8 mm Hg. There was a statistical difference (p < 0.01). Another method is using a current waveform. There is an association between the current and pump flow waves. The current was differentiated and squared to calculate the power of the differentiated current. The normal range of this value was 0.025 +/- 0.029; the abnormal condition was 11.25 +/- 15.13. There was a statistical difference (p < 0.01). The predicted flow estimation method and a sucking detection method were available from intrinsic parameters of the pump and need no sensors. These 2 methods are simple, yet effective and reliable control methods for a rotary blood pump.

Development of an antithrombogenic and antitraumatic blood pump: the Gyro C1E3.

The Gyro C1E3 is a centrifugal blood pump. Its antithrombogenic and antitraumatic blood features were demonstrated by prior studies. Based upon these studies, a mass production model of the C1E3 is becoming commercially available. Therefore, this feasibility study was conducted using the mass production models of the Gyro C1E3 for long-term cardiac assist in ex vivo animal experiments. Five healthy calves were used and 15 pump heads were applied for different time periods (Group 1, 30 days; Group 2, 14 days; Group 3, 10 and 7 days; Group 4, 4 days; and Group 5, 2 days). Activated clotting time (ACT) was kept at 200-250 sec. All five calves demonstrated neither abnormal signs nor abnormal blood examination data throughout the experiment. During necropsy, no thromboembolism was found in any downstream organs. Groups 1-4 showed thrombi inside the pump heads while two pumps in Group 5 had no thrombi formations. Bearing deformation or possible wear did not increase after 2 days of pumping. The C1E3 is capable of long-term assist circulation. However, after 2 days of pumping, careful observation is necessary since thrombi may occur inside the pump when ACT is controlled under 250 sec. During the weaning stage or low flow (under 2 L/min), over 250 sec of ACT is recommended to assure the safety of the patient.

The effect of immunosuppression on aortic dilatation in a rat aneurysm model.

This study was conducted to investigate whether systemic immunosuppression attenuated aortic dilatation in a rat aneurysm model. Sprague-Dawley rats were subjected to elastase infusion of the infrarenal aorta and divided into two groups of 12 rats each. The immunosuppression group (group 1) was given subcutaneous injections of cyclosporine A (5 mg/kg per day), azathioprine (2 mg/kg per day), and methylprednisolone (2 mg/kg per day) from the operative day until postoperative day (POD) 6. An additional subcutaneus injection of cyclophosphamide 30 mg/kg was also given on the operative day. The control group (group 2) was given subcutaneous injections of saline. Relaparotomy was performed on POD 7. After measurement of the aortic diameter, aortography and ultrasonography were performed in three rats from each group, following which the aortas were excised for histologic examination. The aortic diameter was significantly smaller in group 1 (2.58 +/- 0.37 mm) than in group 2 (6.21 +/- 1.74 mm) (P < 0.01) and the aortic lumen was slightly dilated in group 1, whereas it was spherically dilated in group 2. Total loss of elastic tissue was seen in both groups. Inflammatory cell infiltration and collagen fiber fragmentation were noted in group 2, whereas very little inflammatory cell infiltration and well-preserved collagen fibers were seen in group 1. These findings showed that immunosuppression attenuates aortic dilatation, partly by preserving the collagen fibers, in this rat aneurysm model.

Feasibility of a tiny Gyro centrifugal pump as an implantable ventricular assist device.

The Gyro pumps were developed for long-term circulatory support. The first generation Gyro pump (C1E3) achieved 1 month paracorporeal circulatory support in chronic animal experiments; the second generation (PI702) implantable ventricular assist device (VAD) was successful for over 6 months. The objective of the next generation Gyro pump is for use as a long-term totally implantable VAD and for pediatric circulatory support. This tiny Gyro pump (KP101) was fabricated with the same design concept as the other Gyro pumps. The possibility of an implantable VAD was determined after performance and hemolysis test results were compared to those of the other Gyro pumps. The pump housing and impeller were fabricated from polycarbonate with an impeller diameter of 35 mm. The diameter and height of the pump housings are 52.3 mm and 29.9 mm, respectively. At this time, a DC brushless motor drives the KP101, which is the same as that for the C1E3. The pump performance was measured in 37% glycerin water at 37 degrees C. Hemolysis tests were performed utilizing a compact mock loop filled with fresh bovine blood in a left ventricular assist device (LVAD) condition at 37 degrees C. The KP101 achieved the LVAD conditions of 5 L/min and 100 mm Hg at 2,900 rpm; generated 10 L/min against 100 mm Hg at 3,200 rpm; 3 L/min against 90 mm Hg at 2,600 rpm; and 2 L/min against 80 mm Hg at 2,400 rpm. In addition, the pump efficiency during this experiment was 12.5%. The other Gyro pumps. that is, the C1E3, PI601, and PI701, in an LVAD condition require 1,600, 2,000, and 2,000 rpm, respectively. The KP101 produced a normalized index of hemolysis (NIH) value of 0.005 g/100 L. With regard to the NIH, the other Gyro pumps, namely the C1E3, PI601, and PI701 demonstrated 0.0007, 0.0028, and 0.004 g/100 L, respectively. The KP101 produced an acceptable pressure flow curve for a VAD. The NIH value was higher than that of other Gyro pumps, but is in an acceptable range.

A new control method that estimates the backflow in a centrifugal pump.

The rotary blood pump will be widely used in the near future as an implantable left ventricular assist device (LVAD). However, one obstacle for the centrifugal pump is a control method that can maintain an optimum flow rate in a physiological condition. Thus, the object of this study is to develop this optimum control system for the centrifugal pump. If the heart function and pump efficiency are stable, the ratio of the systole current to the the diastole current (S/D) will be a fixed value. However, if the heart function and pump efficiency are unstable, S/D will not be a fixed value. This control system was investigated with a calf that was subjected to an ex vivo LVAD study. The LVAD was a Gyro C1E3 centrifugal pump. The pump flow rate was changed to 1.5, 3.5, 5.2, and 6.2 L/min. According to the changes of the pump flow rates, the S/D values were 1.01 +/- 0.01, 1.06 +/- 0.05, 1.03 +/- 0.01, and 1.03 +/- 0.01, respectively. There was no statistical difference among the 3 groups. In a separate experiment, the backflow condition S/D was 1.88 +/- 0.6, and the normal condition S/D was 1.35 +/- 0.5. There was a statistical difference between the 2 groups. The results of this study suggest that S/D is not influenced by the pump flow rate. However, the S/D was changed when the pump was in a backflow condition. This method will be useful in controlling a centrifugal pump requiring only electrical current information.

Shortened elastase infusion time in the elastase-induced rat aneurysm model.

OBJECTIVE: This study was performed to determine whether it would be possible to shorten the elastase infusion time in the elastase-induced rat aneurysm model. METHODS: The abdominal aortas of 76 male Sprague-Dawley rats were dissected and infused for 10, 20, 30, 60, or 120 min with a solution containing 14 U of elastase or for 30 min with saline solution (control). After infusion, the rats were evaluated every day for calculation of the mortality rate. On day 7, the surviving rats underwent laparotomy. The diameter of the aorta was measured, and the aortic tissue was excised for histologic examination. RESULTS: There were no deaths among the rats infused for 10, 20, or 30 min. The mortality rate was 64% in the 60-min and 90% in 120-min groups. There were no significant differences in the diameters of the 30-min saline-infused aortas and the 10- or 20-min elastase-infused aortas. The diameter of the 30-min elastase-infused aortas (6.2 +/- 2.1 mm) was significantly larger (P < 0.01) than the diameters of the 10- and 20-min elastase-infused aortas. There were no significant differences between the 30-min and the 60- and 120-min elastase-infused aortas. Microscopically, there was total or subtotal loss of elastic tissue and marked inflammatory cell infiltration of the aortic wall in the 30-, 60-, and 120-min elastase-infused aortas. CONCLUSIONS: It is possible to shorten the elastase infusion time from 120 to 30 min in the elastase-induced rat aneurysm model. This shortening of infusion time reduces the experimental time and mortality rate.