AC Electromagnetic Field Controls the Biofilms on the Glass Surface by Escherichia coli & Staphylococcus epidermidis Inhibition Effect.

Biofilms, mainly comprised of bacteria, form on materials' surfaces due to bacterial activity. They are generally composed of water, extracellular polymeric substances (polysaccharides, proteins, nucleic acids, and lipids), and bacteria. Some bacteria that form biofilms cause periodontal disease, corrosion of the metal materials that make up drains, and slippage. Inside of a biofilm is an environment conducive to the growth and propagation of bacteria. Problems with biofilms include the inability of disinfectants and antibiotics to act on them. Therefore, we have investigated the potential application of alternating electromagnetic fields for biofilm control. We obtained exciting results using various materials' specimens and frequency conditions. Through these studies, we gradually understood that the combination of the type of bacteria, the kind of material, and the application of an electromagnetic field with various low frequencies (4 kHz-12 kHz) changes the circumstances of the onset of the biofilm suppression effect. In this study, relatively high frequencies (20 and 30 kHz) were applied to biofilms caused by Escherichia coli (E. coli) and Staphylococcus epidermidis (S. epidermidis), and quantitative evaluation was performed using staining methods. The sample surfaces were analyzed by Raman spectroscopy using a Laser Raman spectrometer to confirm the presence of biofilms on the surface.

Quantitative Analyses of Biofilm by Using Crystal Violet Staining and Optical Reflection.

Biofilms have caused many problems, not only in the industrial fields, but also in our daily lives. Therefore, it is important for us to control them by evaluating them properly. There are many instrumental analytical methods available for evaluating formed biofilm qualitatively. These methods include the use of Raman spectroscopy and various microscopes (optical microscopes, confocal laser microscopes, scanning electron microscopes, transmission electron microscopes, atomic force microscopes, etc.). On the other hand, there are some biological methods, such as staining, gene analyses, etc. From the practical viewpoint, staining methods seem to be the best due to various reasons. Therefore, we focused on the staining method that used a crystal violet solution. In the previous study, we devised an evaluation process for biofilms using a color meter to analyze the various staining situations. However, this method was complicated and expensive for practical engineers. For this experiment, we investigated the process of using regular photos that were quantified without any instruments except for digitized cameras. Digitized cameras were used to compare the results. As a result, we confirmed that the absolute values were different for both cases, respectively. However, the tendency of changes was the same. Therefore, we plan to utilize the changes before and after biofilm formation as indicators for the future.

Proposal for Some Affordable Laboratory Biofilm Reactors and Their Critical Evaluations from Practical Viewpoints.

Biofilms are a result of bacterial activities and are found everywhere. They often form on metal surfaces and on the surfaces of polymeric compounds. Biofilms are sticky and mostly consist of water. They have a strong resistance to antimicrobial agents and can cause serious problems for modern medicine and industry. Biofilms are composed of extracellular polymeric substances (EPS) such as polysaccharides produced from bacterial cells and are dominated by water at the initial stage. In a series of experiments, using Escherichia coli, we developed three types of laboratory biofilm reactors (LBR) to simulate biofilm formation. For the first trial, we used a rotary type of biofilm reactor for stirring. For the next trial, we tried another rotary type of reactor where the circular plate holding specimens was rotated. Finally, a circular laboratory biofilm reactor was used. Biofilms were evaluated by using a crystal violet staining method and by using Raman spectroscopy. Additionally, they were compared to each other from the practical (industrial) viewpoints. The third type was the best to form biofilms in a short period. However, the first and second were better from the viewpoint of "ease of use". All of these have their own advantages and disadvantages, respectively. Therefore, they should be properly selected and used for specific and appropriate purposes in the future.

Impedance Characteristics of Monolayer and Bilayer Graphene Films with Biofilm Formation and Growth.

Biofilms are the result of bacterial activity. When the number of bacteria (attached to materials' surfaces) reaches a certain threshold value, then the bacteria simultaneously excrete organic polymers (EPS: extracellular polymeric substances). These sticky polymers encase and protect the bacteria. They are called biofilms and contain about 80% water. Other components of biofilm include polymeric carbon compounds such as polysaccharides and bacteria. It is well-known that biofilms cause various medical and hygiene problems. Therefore, it is important to have a sensor that can detect biofilms to solve such problems. Graphene is a single-atom-thick sheet in which carbon atoms are connected in a hexagonal shape like a honeycomb. Carbon compounds generally bond easily to graphene. Therefore, it is highly possible that graphene could serve as a sensor to monitor biofilm formation and growth. In our previous study, monolayer graphene was prepared on a glass substrate by the chemical vapor deposition (CVD) method. Its biofilm forming ability was compared with that of graphite. As a result, the CVD graphene film had the higher sensitivity for biofilm formation. However, the monolayer graphene has a mechanical disadvantage when used as a biofilm sensor. Therefore, for this new research project, we prepared bilayer graphene with high mechanical strength by using the CVD process on copper substrates. For these specimens, we measured the capacitance component of the specimens' impedance. In addition, we have included a discussion about the possibility of applying them as future sensors for monitoring biofilm formation and growth.

Measurement of hemodynamic changes with the axial flow blood pump installed in descending aorta.

We have developed various axial flow blood pumps to realize the concept of the Valvo pump, and we have studied hemodynamic changes under cardiac assistance using an axial flow blood pump in series with the natural heart. In this study, we measured hemodynamic changes of not only systemic circulation but also cerebral circulation and coronary circulation under cardiac support using our latest axial flow blood pump placed in the descending aorta in an acute animal experiment. The axial flow blood pump was installed at the thoracic descending aorta through a left thoracotomy of a goat (43.8 kg, female). When the pump was on, the aortic pressure and aortic flow downstream of the pump increased with preservation of pulsatilities. The pressure drop upstream of the pump caused reduction of afterload pressure, and it may lead to reduction of left ventricular wall stress. However, cerebral blood flow and coronary blood flow were decreased when the pump was on. The axial flow blood pump enables more effective blood perfusion into systemic circulation, but it has the potential risk of blood perfusion disturbance into cerebral circulation and coronary circulation. The results indicate that the position before the coronary ostia might be suitable for implantation of the axial flow blood pump in series with the natural heart to avoid blood perfusion disturbances.

Sensorless cardiac phase detection for synchronized control of ventricular assist devices using nonlinear kernel regression model.

Recently, driving methods for synchronizing ventricular assist devices (VADs) with heart rhythm of patients suffering from severe heart failure have been receiving attention. Most of the conventional methods require implanting a sensor for measurement of a signal, such as electrocardiogram, to achieve synchronization. In general, implanting sensors into the cardiovascular system of the patients is undesirable in clinical situations. The objective of this study was to extract the heartbeat component without any additional sensors, and to synchronize the rotational speed of the VAD with this component. Although signals from the VAD such as the consumption current and the rotational speed are affected by heartbeat, these raw signals cannot be utilized directly in the heartbeat synchronization control methods because they are changed by not only the effect of heartbeat but also the change in the rotational speed itself. In this study, a nonlinear kernel regression model was adopted to estimate the instantaneous rotational speed from the raw signals. The heartbeat component was extracted by computing the estimation error of the model with parameters determined by using the signals when there was no effect of heartbeat. Validations were conducted on a mock circulatory system, and the heartbeat component was extracted well by the proposed method. Also, heartbeat synchronization control was achieved without any additional sensors in the test environment.

Hemodynamics of a functional centrifugal-flow total artificial heart with functional atrial contraction in goats.

Implantation of a total artificial heart (TAH) is one of the therapeutic options for the treatment of patients with end-stage biventricular heart failure. There is no report on the hemodynamics of the functional centrifugal-flow TAH with functional atrial contraction (fCFTAH). We evaluated the effects of pulsatile flow by atrial contraction in acute animal models. The goats received fCFTAH that we created from two centrifugal-flow ventricular assist devices. Some hemodynamic parameters maintained acceptable levels: heart rate 115.5 ± 26.3 bpm, aortic pressure 83.5 ± 10.1 mmHg, left atrial pressure 18.0 ± 5.9 mmHg, pulmonary pressure 28.5 ± 9.7 mmHg, right atrial pressure 13.6 ± 5.2 mmHg, pump flow 4.0 ± 1.1 L/min (left) 3.9 ± 1.1 L/min (right), and cardiac index 2.13 ± 0.14 L/min/m(2). fCFTAH with atrial contraction was able to maintain the TAH circulation by forming a pulsatile flow in acute animal experiments. Taking the left and right flow rate balance using the low internal pressure loss of the VAD pumps may be easier than by other pumps having considerable internal pressure loss. We showed that the remnant atrial contraction effected the flow rate change of the centrifugal pump, and the atrial contraction waves reflected the heart rate. These results indicate that remnant atria had the possibility to preserve autonomic function in fCFTAH. We may control fCFTAH by reflecting the autonomic function, which is estimated with the flow rate change of the centrifugal pump.

Engineering based assessment for a shape design of a pediatric ePTFE pulmonary conduit valve.

The authors examined the hemodynamic characteristics of expanded polytetrafluoroethylene (ePTFE) pulmonary valved conduits quantitatively by our originally developed pediatric pulmonary mechanical circulatory system, in order to suggest the optimal shape design. The system consisted of pneumatically driven right atrium and ventricle model, a pulmonary valve chamber, and elastic pulmonary compliance model with peripheral vascular resistance units, a venous reservoir. We employed two different types of ePTFE valve and evaluated the relationship between the leaflets motion and hemodynamic characteristics by using a high-speed video camera. As a result, we successfully reproduced hemodynamic simulations in our pediatric pulmonary mock system. We confirmed that the presence of bulging sinuses in the pulmonary valved conduit reduced the transvalvular energy loss and increased the valve opening area during systolic period. Our engineering-based in vitro analysis could be useful for proposing a shape design optimization of sophisticated pediatric ePTFE pulmonary valve.

Initial Acute Animal Experiment Using a New Miniature Axial Flow Pump in Series With the Natural Heart.

We have advocated an axial flow blood pump called "valvo pump" that is implanted at the aortic valve position, and we have developed axial flow blood pumps to realize the concept of the valvo pump. The latest model of the axial flow blood pump mainly consists of a stator, a directly driven impeller, and a hydrodynamic bearing. The axial flow blood pump has a diameter of 33 mm and length of 74 mm, and the length of anatomical occupation is 33 mm. The axial flow blood pump is anastomosed to the aorta with polytetrafluoroethylene (PTFE) cuffs worn on the inflow and outflow ports. Dp-Q curves of the axial flow blood pump are flatter than those of ordinary axial flow pumps, and pump outflow of 5 L/min was obtained against a pressure difference of 50 mm Hg at a rotational speed of 9000 rpm in vitro. The axial flow blood pump was installed in a goat by anastomosing with the thoracic descending aorta using PTFE cuffs, and it was rotated at a rotational speed of 8000 rpm. Unlike in case of the ventricular assistance in parallel with the natural heart, pulsatilities of aortic pressure and aortic flow were preserved even when the pump was on, and mean aortic flow was increased by 1.5 L/min with increase in mean aortic pressure of 30 mm Hg. In conclusion, circulatory assistance in series with the natural heart using the axial flow blood pump was able to improve hemodynamic pulsatility, and it would contribute to improvement of end-organ circulation. .

Development of a thermodynamic control system for the Fontan circulation pulsation device using shape memory alloy fibers.

The Fontan procedure is one of the common surgical treatments for circulatory reconstruction in pediatric patients with congenital heart disease. In Fontan circulation, low pulsatility may induce localized lung ischemia and may impair the development of pulmonary peripheral endothelial cells. To promote pulmonary circulation in Fontan circulation, we have been developing a pediatric pulmonary circulatory pulsation device using shape memory alloy fibers attached from the outside of total cavopulmonary connection. In this study, we developed a new thermal control system for the device and examined its functions. We mounted on the device 16 fibers connected in parallel around an ePTFE graft circumferentially. To provide optimized contraction, we designed the new thermal control system. The system consisted of a thermistor, a pressure sensor, and a regulator that was controlled by the adaptive thermodynamic transfer functions. We monitored the parameters and calculated heat transfer function as well as pressure distribution on the graft surface. Then we examined and compared the dynamic contractile pressure and changes in surface temperature. As a result, by the application of the control based on the new feedback system analysis, the circumferential contractile pressure increased by 35%. The adaptive thermodynamic regulation was useful for the selection of alternative thresholds of the surface temperature of the graft. The system could achieve effective contraction for the pulsatile flow generation by the device.

Fundamental analysis and development of the current and voltage control method by changing the driving frequency for the transcutaneous energy transmission system.

We have been developing transcutaneous energy transmission system (TETS) for a ventricular assist device, shape memory alloy (SMA) fibered artificial organs and so on, the system has high efficiency and a compact size. In this paper, we summarize the development, design method and characteristics of the TETS. New control methods for stabilizing output voltage or current of the TETS are proposed. These methods are primary side, are outside of the body, not depending on a communication system from the inside the body. Basically, the TETS operates at the fixed frequency with a suitable compensation capacitor so that the internal impedance is minimalized and a flat load characteristic is obtained. However, when the coil shifted from the optimal position, the coupling factor changes and the output is fluctuated. TETS has a resonant property; its output can be controlled by changing the driving frequency. The continuous current to continuous voltage driving method was implemented by changing driving frequency and setting of limitation of low side frequency. This method is useful for battery charging system for electrically driven artificial hearts and also useful for SMA fibered artificial organs which need intermittent high peak power comsumption. In this system, the internal storage capacitor is charged slowly while the fibers are turned off and discharge the energy when the fibers are turned on. We examined the effect of the system. It was found that the size and maximum output of the TETS would able to be reduced.

Discrimination ability and reproducibility of a new index reflecting autonomic nervous function based on pulsatile amplitude of photoplethysmography.

A new physiological index (µ(PA)) is proposed to represent the autonomic nervous system (ANS) function. The index µ(PA) is defined as the natural logarithm of the ratio between two different frequency components of the pulsatile amplitude of the photoplethysmogram (PPG) signal. The discrimination ability and the reproducibility of µ(PA) have been compared with other traditional ANS indices. In the experiment, the electrocardiogram, the PPG and continuous blood pressure were measured in 59 healthy young subjects (age 25.7 ± 6.3) and 86 healthy elderly subjects (age 70.2 ± 4.1) at rest. The discrimination ability and the reproducibility were evaluated by Cohen's d between young and elderly groups and by the interclass correlation coefficient, respectively. The results showed that the elderly subjects were significantly (p<;0.001) lower than young subjects in µ(PA) and a few traditional indices introduced to be compared with µ(PA). Therefore, it suggests that µ(PA) is associated with the decrease in the ANS function accompanied by aging. Moreover, it showed that the discrimination ability and the reproducibility of the proposed index are comparable or larger than those of traditional indices. The proposed index based on the PPG signal will be applied to tele-healthcare systems for monitoring people's health in daily life in combination with the ratio of the standard deviation of the R-R intervals to their average value (CVRR).

Hemodynamic effects of pressure-volume relation in the atrial contraction model on the total artificial heart using centrifugal blood pumps.

Hemodynamic effects of atrial contraction with centrifugal pump type total artificial heart is unknown. In this study, we simulated an atrial contraction in a mock model. By the driving condition with higher pressure in the mock atrial model, the load during atrial contraction increased. Based on these findings, we examined atrial contraction in the animal using adult goats. Prior to the measurement, we installed a centrifugal-type ventricular assist device (VADs), and then clamped both ventricles. We measured the hemodynamic data without ventricular contractile functions in order to obtain the effect of atrial contraction on hemodynamics under the condition of the total artificial heart (TAH) circulatory support model. We could estimate the heart rate by revolution number and voltage of pumps. There might be a possibility that we could regulate autonomic nervous response with the control of cardiac output.

Expansion capsules for diet control with artificial organ technology.

When we consider the medical economy, the Obesity is one of the leading preventable causes of death worldwide. However, a lot of previous scientific papers reported that 95% of obesity patients would not be able to control their weight by the diet. The surgical operation has been considered to the subjects with severe obesity. But, there is a possibility of complication or comorbidity in surgical operation. Tohoku University started to develop the expanding capsule with transcutaneous energy transmission system (TETS) having the same effect as the surgical operation. The capsule in the stomach will expand mechanically by energy transmission from outsides of the body, when the obesity patients will felt hungry. Small linear drive with folding umbrella type actuator would enable us the expansion of the diet capsules. Satisfactory characteristic of the energy transmission was obtained by the trial model of TETS during animal experiments. Animal experiments with healthy adult goats enabled us the evaluation of the inner stomach pressure time series changes, and feasibility study. Double blind test of the expanding capsule is now under planning. If the expanding capsule diet control system will be embodied, it becomes the gospel of the obese subject.

Transcutaneous communication system using the human body as conductive medium: influence of transmission data current on the heart.

We developed a new transcutaneous communication system (TCS) that uses the human body as a conductive medium for monitoring and controlling artificial hearts and other artificial organs in the body.In this study, the physiological effect of data current discharged into the body during data transmission was evaluated by an animal experiment using a goat. The external and internal units of the new TCS each mainly consist of a data transmitter and a data receiver. The data transmitter has an amplitude shift keying (ASK) modulator (carrier frequencies: 4 and 10 MHz) and an electrode.The internal unit of the TCS was fixed on the pericardium and the external unit was placed on the left ear, and each transmitter discharged an ASK-modulated current of 7 mA (RMS) into the conscious goat. The TCS was able to transmit data for 4 weeks under full duplex communication with a transmission rate of 115 kbps. On the 28th postoperative day, an electrocardiogram was measured during data transmission. Cardiac rhythm and waveform of the electrocardiogram were not changed before and during bidirectional data transmission. Also, no adverse effect on the heart was observed by autopsy.

A new transcutaneous bidirectional communication for monitoring implanted artificial heart using the human body as a conductive medium.

A transcutaneous communication system (TCS) is a key technology for monitoring and controlling artificial hearts and other artificial organs in the body. In this study, we developed a new TCS that uses the human body as a conductive medium. Direct data exchange provides a higher level of communication security compared to that of wireless methods without physical constraints such as an external wire. The external and internal units of the new TCS each consist mainly of a data transmitter and a data receiver. The data transmitter has an amplitude shift keying (ASK) modulator (carrier frequencies: 4 and 10 MHz) and an electrode. The ASK-modulated data current is led into the body through the electrode, and it flows back to the energy source through the body, the data receiver, and the earth ground that includes all conductors and dielectrics in the environment that are in close proximity to the patient. Performance of the TCS was evaluated by a communication test on the surface of the human body and in an animal experiment using a goat. The TCS was able to transmit data concurrently for 4 weeks between everywhere on the surface of the body and everywhere inside the body under full-duplex communication at a transmission rate of 115 kbps. The power consumption of each TCS unit was 125 mW with an ASK-modulated current of 7 mA (root-mean-square). While further study is required to secure its safety, the newly developed TCS has promise to be a next-generation transcutaneous communication device.

New implantable therapeutic device for the control of an atrial fibrillation attack using the Peltier element.

For the development of the new therapeutic device for the atrial fibrillation, implantable cooling device using Peltier element was developed in this study. An implantable cooling device had been consisted from Peltier element with transcutaneous energy transmission system (TETS). 1st coil can be contacted from outside of the body, when the patients will feel palpitation. Electrical current will be induced to the implanted 2nd coil. Peltier element will able to cool the surface of the atrium. For the confirmation of the effect of the cooling device, trial manufacture model was developed. Animal experiments using six healthy adult goats after animal ethical committee allowance was carried out. Fourth intercostals space had been opened after anesthesia inhalation, and various sensors had been inserted. AF was induced by the electrical current with battery. As the results, AF had been recovered to the normal sinus rhythm after cooling in all six goats. So, this cooling system for the control of AF showed evident effect in these experiments. Smaller size cooling device has been under development aiming at totally implantable type. Catheter type cooling device for the insertion by the use of fiber-scope type is now under planning for the clinical application. This new type device may be able to become good news for the patients with uncontrollable AF.

Use of in vivo insert molding to form a jellyfish valve leaflet.

We developed an in vivo insert molding technique to form tissue-derived biomaterials into the desired shape, and with sufficient strength and durability, for use in artificial organs. Molds of acrylic resin with inserted velour cloth were implanted under the skin of goats to form a circular leaflet for a jellyfish valve. The valve leaflets were successfully produced in the molds after 17-60 days. Dense connective tissue covered the velour cloth, and loose connective tissue was formed within it. Tissue was radially formed from the hole in the mold. The tissue was simultaneously formed and shrunk. It is necessary to increase the connected portion between the tissue inside and outside the mold so that the tissue can completely cover the inserted materials without shrinkage.

A nonpulsatile total artificial heart with 1/R control.

A total artificial heart (TAH) using continuous flow pumps is promising for size reduction of the device; however, the role of pulsatility in TAHs has been a subject of great debate. Additionally, it is unclear whether, in a nonpulsatile TAH, a physiological control method such as 1/R control can keep the experimental animal in good condition. To realize a nonpulsatile TAH with 1/R control, the artificial valves were removed from undulation pump total artificial hearts (UPTAHs), which can produce both pulsatile and nonpulsatile flows using a single device. The UPTAHs were implanted into 18 goats, and 4 goats survived for more than 1 month. Three weeks of long-term nonpulsatile TAH operation could be tested in the goat that survived for 72 days, and it was proved that 1/R control is possible not only with a pulsatile TAH but also with a nonpulsatile TAH. The general condition of the goat and its organ function did not change on the application of nonpulsatile mode. Cardiac output and arterial pressure changed with the condition of the goat in pulsatile and also in nonpulsatile modes, and the changes seemed almost identical. However, the sucking effect of the atria was very significant in nonpulsatile mode, resulting in hemolysis. Therefore, nonpulsatile TAHs under 1/R control are considered to be inadequate unless some pulsatility can be introduced to avoid fatal sucking effects and to ensure sufficient inflow. During nonpulsatile operation, regular fluctuations were sometimes found in the aortic pressure, and these were caused by the periodic sucking effect in the left atrium that was possibly influenced by respiratory changes.