Liquid-Liquid Phase Separation in the Presence of Macromolecular Crowding and State-dependent Kinetics.

Biomolecular condensates formed via liquid-liquid phase separation (LLPS) are increasingly being shown to play major roles in cellular self-organization dynamics in health and disease. It is well established that macromolecular crowding has a profound impact on protein interactions, particularly those that lead to LLPS. Although synthetic crowding agents are used during in vitro LLPS experiments, they are considerably different from the highly crowded nucleo-/cytoplasm and the effects of in vivo crowding remain poorly understood. In this work, we applied computational modeling to investigate the effects of macromolecular crowding on LLPS. To include biologically relevant LLPS dynamics, we extended the conventional Cahn-Hilliard model for phase separation by coupling it to experimentally derived macromolecular crowding dynamics and state-dependent reaction kinetics. Through extensive field-theoretic computer simulations, we show that the inclusion of macromolecular crowding results in late-stage coarsening and the stabilization of relatively smaller condensates. At a high crowding concentration, there is an accelerated growth and late-stage arrest of droplet formation, effectively resulting in anomalous labyrinthine morphologies akin to protein gelation observed in experiments. These results not only elucidate the crowder effects observed in experiments, but also highlight the importance of including state-dependent kinetics in LLPS models, and may help in designing further experiments to probe the intricate roles played by LLPS in self-organization dynamics of cells.

H2O2-Responsive Antioxidant Nanoparticle Attenuates Whole Body Ischemia/Reperfusion-Induced Multi-Organ Damages.

Mortality and morbidity after cardiac arrest remain high due to ischemia/reperfusion (I/R) injury causing multi-organ damages, even after successful return of spontaneous circulation. We previously generated H2O2-activatable antioxidant nanoparticles formulated with copolyoxalate containing vanillyl alcohol (PVAX) to prevent I/R injury. In this study, we examined whether PVAX could effectively reduce organ damages in a rat model of whole-body ischemia/reperfusion injury (WBIR). To induce a cardiac arrest, 70µl/100 g body weight of 1 mmol/l potassium chloride was administered via the jugular venous catheter. The animals in both the vehicle and PVAX-treated groups had similar baseline blood pressure. After 5.5 minutes of cardiac arrest, animals were resuscitated via intravenous epinephrine followed by chest compressions. PVAX or vehicle was injected after the spontaneous recovery of blood pressure was noted, followed by the same dose of second injection 10 minutes later. After 24 hours, multiple organs were harvested for pathological, biochemical, molecular analyses. No significant difference on the restoration of spontaneous circulation was observed between vehicle and PVAX groups. Analysis of organs harvested 24 hours post procedure showed that whole body I/R significantly increased reactive oxygen species (ROS) generation, inflammatory markers, and apoptosis in multiple organs (heart, brain, and kidney). PVAX treatment effectively blocked ROS generation, reduced the elevation of pro-inflammatory cytokines, and decreased apoptosis in these organs. Taken together, our results suggest that PVAX has potent protective effect against WBIR induced multi-organ injury, possibly by blocking ROS-mediated cell damage.

Assessment of Electrocardiogram Rhythms by GoogLeNet Deep Neural Network Architecture.

The aim of this study is to design GoogLeNet deep neural network architecture by expanding the kernel size of the inception layer and combining the convolution layers to classify the electrocardiogram (ECG) beats into a normal sinus rhythm, premature ventricular contraction, atrial premature contraction, and right/left bundle branch block arrhythmia. Based on testing MIT-BIH arrhythmia benchmark databases, the scope of training/test ECG data was configured by covering at least three and seven R-peak features, and the proposed extended-GoogLeNet architecture can classify five distinct heartbeats; normal sinus rhythm (NSR), premature ventricular contraction (PVC), atrial premature contraction (APC), right bundle branch block (RBBB), and left bundle brunch block(LBBB), with an accuracy of 95.94%, an error rate of 4.06%, a maximum sensitivity of 96.9%, and a maximum positive predictive value of 95.7% for judging a normal or an abnormal beat with considering three ECG segments; an accuracy of 98.31%, a sensitivity of 88.75%, a specificity of 99.4%, and a positive predictive value of 94.4% for classifying APC from NSR, PVC, APC beats, whereas the error rate for misclassifying APC beat was relative low at 6.32%, compared with previous research efforts.

Targeting Oxidative Stress Using Nanoparticles as a Theranostic Strategy for Cardiovascular Diseases.

SIGNIFICANCE: Nanomedicine is an application of nanotechnology that provides solutions to unmet medical challenges. The unique features of nanoparticles, such as their small size, modifiable components, and diverse functionality, make them attractive and suitable materials for novel diagnostic, therapeutic, or theranostic applications. Cardiovascular diseases (CVDs) are the major cause of noncommunicable illness in both developing and developed countries. Nanomedicine offers novel theranostic options for the treatment of CVDs. Recent Advances: Many innovative nanoparticles to target reactive oxygen species (ROS) have been developed. In this article, we review the characteristics of nanoparticles that are responsive to ROS, their limitations, and their potential clinical uses. Significant advances made in diagnosis of atherosclerosis and treatment of acute coronary syndrome using nanoparticles are discussed. CRITICAL ISSUES: Although there is a tremendous potential for the nanoparticle applications in medicine, their safety should be considered while using in humans. We discuss the challenges that may be encountered with some of the innovative nanoparticles used in CVDs. FUTURE DIRECTIONS: The unique properties of nanoparticles offer novel diagnostic tool and potential therapeutic strategies. However, nanomedicine is still in its infancy, and further in-depth studies are needed before wide clinical application is achieved.

Anticancer activity of biologically synthesized silver and gold nanoparticles on mouse myoblast cancer cells and their toxicity against embryonic zebrafish.

The aim of this study was to evaluate the anticancer activity of bioinspired silver nanoparticles (AgNPs) and gold nanoparticles (AuNPs) against mouse myoblast cancer cells (C2C12). Both AgNPs and AuNPs were biologically synthesized using Spinacia oleracea Linn., aqueous leaves extract. UV-Vis. spectrophotometer, high resolution-transmission electron microscopy (HR-TEM), field emission-scanning electron microscopy (FE-SEM) and X-ray diffraction (XRD) studies supported the successful synthesis of AgNPs and AuNPs. Both these NPs have shown cytotoxicity against C2C12 cells even at very low concentration (5µg/mL). Acridine orange/Ethidium bromide (AO/EB) dual staining confirmed the apoptotic morphological features. The levels of caspase enzymes (caspase-3 and caspase-7) were significantly up-regulated in NPs treated myoblast cells than the plant extract. Furthermore, in zebrafish embryo toxicity study, AgNPs showed 100% mortality at 3µg/mL concentration while AuNPs exhibited the same at much higher concentration (300mg/mL). Taken together, these results provide a preliminary guidance for the development of biomaterials based drugs to fight against the fatal diseases for example cancer.

Removal of baseline wandering in ECG signal by improved detrending method.

The very first step to process electrocardiogram (ECG) signal is to eliminate baseline wandering interference that is usually caused by electrode-skin impedance mismatch, motion artifacts due to a patient's body moment or respiratory breathing. A new method is thus suggested to remove baseline wandering in ECG by improving the detrending method that was originally proposed for eliminating slow non-stationary trends from heart rate variability (HRV). In our proposed method, a global trend is estimated in terms of baseline wandering by merging the local trend based on an ECG segment that represents a part of the ECG signal. The experimental results show that the improved detrending method can efficiently resolve baseline wandering without distorting any morphological characteristic embedded in the ECG signal in no time delay manner.

Enhancement of objects in photoacoustic tomography using selective filtering.

Here we developed a real-time photoacoustic tomography (PAT) imaging acquisition device based on the linear array transducer utilized on ultrasonic devices. Also, we produced a phantom including diverse contrast media and acquired PAT imaging as the light source wavelength was changing to see if the contrast media reacted. Indocyanine green showed the highest reaction around the 800-nm band, methylene blue demonstrated the same in the 750-nm band, and gold nanoparticle showed the same in the 700-nm band. However, in the case of superparamagnetic iron oxide, we observed not reaction within the wavelength bands used herein to obtain imaging. Moreover, we applied selective filtering to the acquired PAT imaging to remove noise from around and reinforce the object's area. Consequentially, we could see the object area in the imaging was effectively detected and the image noise was removed.

Real-time photoacoustic tomograpghy using linear array probe and detection of line structure using Hough transform.

A real-time photoacoustic tomography (PAT) system is developed using a linear array probe and phantom images are acquired with a pattern of line structure. Moreover, it is attempted to detect line structures from the acquired images by Hough transform. This effort leads to the measurement of a process of magenta passing through a tube and acquisition of images at a speed of about 2 frame/sec. Besides, it is confirmed that the Hough transform applied on the acquired PAT images has the detection rate of about 50% for delineating a line structure.

Reactive oxygen species-activated nanomaterials as theranostic agents.

Reactive oxygen species (ROS) are generated from the endogenous oxidative metabolism or from exogenous pro-oxidant exposure. Oxidative stress occurs when there is excessive production of ROS, outweighing the antioxidant defense mechanisms which may lead to disease states. Hydrogen peroxide (H2O2) is one of the most abundant and stable forms of ROS, implicated in inflammation, cellular dysfunction and apoptosis, which ultimately lead to tissue and organ damage. This review is an overview of the role of ROS in different diseases. We will also examine ROS-activated nanomaterials with emphasis on hydrogen peroxide, and their potential medical implications. Further development of the biocompatible, stimuli-activated agent responding to disease causing oxidative stress, may lead to a promising clinical use.

Patterned thin metal film for the lateral resolution measurement of photoacoustic tomography.

BACKGROUND: Image quality assessment method of photoacoustic tomography has not been completely standardized yet. Due to the combined nature of photonic signal generation and ultrasonic signal transmission in biological tissue, neither optical nor ultrasonic traditional methods can be used without modification. An optical resolution measurement technique was investigated for its feasibility for resolution measurement of photoacoustic tomography. METHODS: A patterned thin metal film deposited on silica glass provides high contrast in optical imaging due to high reflectivity from the metal film and high transmission from the glass. It provides high contrast when it is used for photoacoustic tomography because thin metal film can absorb pulsed laser energy. An US Air Force 1951 resolution target was used to generate patterned photoacoustic signal to measure the lateral resolution. Transducer with 2.25 MHz bandwidth and a sample submerged in water and gelatinous block were tested for lateral resolution measurement. RESULTS: Photoacoustic signal generated from a thin metal film deposited on a glass can propagate along the surface or through the surrounding medium. First, a series of experiments with tilted sample confirmed that the measured photoacoustic signal is what is propagating through the medium. Lateral resolution of the photoacoustic tomography system was successfully measured for water and gelatinous block as media: 0.33 mm and 0.35 mm in water and gelatinous material, respectively, when 2.25 MHz transducer was used. Chicken embryo was tested for biomedical applications. CONCLUSIONS: A patterned thin metal film sample was tested for its feasibility of measuring lateral resolution of a photoacoustic tomography system. Lateral resolutions in water and gelatinous material were successfully measured using the proposed method. Measured resolutions agreed well with theoretical values.

Availability of a newly devised ambulatory urodynamics monitoring system based on personal device assistance in patients with spinal cord injury.

Conventional urodynamics systems have been widely used for the assessment of bladder functions. However, they have some drawbacks due to the unfamiliar circumstances for the patient, restrictive position during the test, expense and immovability of the instrument as well as the unphysiological filling of the bladder. To mitigate these problems, we developed a fully ambulatory urodynamics monitoring system, which enables the abdominal pressure to be measured in a non-invasive manner, as well as the manual recording of various events such as the bladder sensations or leakage of urine. Conventional (CMG) and furosemide-stimulated filling cystometry (FCMG) were performed for 28 patients with neurogenic bladders caused by spinal cord injury (24 males and 4 females, age: 49.4 ± 13.9 years, BMI: 23.5 ± 2.4). There were high correlation coefficients (r=0.97 ± 0.02) between the clinical parameters measured by the conventional rectal catheter and those measured by our non-invasive algorithm in the FCMG studies. Also, 10 of the patients (36%) were diagnosed as having different reflexibility of the bladder between conventional CMG and FCMG (p<0.05). In the patients with detrusor overactivity, the average volume and detrusor pressure at bladder sensation in FCMG were lower than those in CMG, while the average compliance was higher (p<0.05). In the patients with areflexic bladders, the number of patients with detrusor overactivity was higher in FCMG and leakage was observed more frequently. These results showed that our system could be a useful additional tool in the clinical assessment of patients in which conventional cystometry failed to explain their symptoms.

An acoustical evaluation of knee sound for non-invasive screening and early detection of articular pathology.

Knee sound signals generated by knee movement are sometimes associated with degeneration of the knee joint surface and such sounds may be a useful index for early disease. In this study, we detected the acoustical parameters, such as the fundamental frequency (F0), mean amplitude of the pitches, and jitter and shimmer of knee sounds, and compared them according to the pathological conditions. Six normal subjects (4 males and 2 females, age: 28.3 ± 2.3 years) and 11 patients with knee problems were enrolled. The patients were divided into the 1st patient group (5 males and 1 female, age: 30.2 ± 10.3 years) with ruptured wounds of the meniscus and 2nd patient group (2 males and 3 females, age: 42.1 ± 16.2 years) with osteoarthritis. The mean values of F0, jitter and shimmer of the 2nd patient group were larger than those of the normal group, whereas those of the 1st patient group were smaller (p < 0.05). Also, the F0 and jitter in the standing position were larger than those in the sitting position in both the 1st and 2nd patient groups (p < 0.05). These results showed good potential for the non-invasive diagnosis and early detection of articular pathologies via an auscultation.

Non-invasive algorithm for bowel motility estimation using a back-propagation neural network model of bowel sounds.

BACKGROUND: Radiological scoring methods such as colon transit time (CTT) have been widely used for the assessment of bowel motility. However, these radiograph-based methods need cumbersome radiological instruments and their frequent exposure to radiation. Therefore, a non-invasive estimation algorithm of bowel motility, based on a back-propagation neural network (BPNN) model of bowel sounds (BS) obtained by an auscultation, was devised. METHODS: Twelve healthy males (age: 24.8 ± 2.7 years) and 6 patients with spinal cord injury (6 males, age: 55.3 ± 7.1 years) were examined. BS signals generated during the digestive process were recorded from 3 colonic segments (ascending, descending and sigmoid colon), and then, the acoustical features (jitter and shimmer) of the individual BS segment were obtained. Only 6 features (J1, 3, J3, 3, S1, 2, S2, 1, S2, 2, S3, 2), which are highly correlated to the CTTs measured by the conventional method, were used as the features of the input vector for the BPNN. RESULTS: As a results, both the jitters and shimmers of the normal subjects were relatively higher than those of the patients, whereas the CTTs of the normal subjects were relatively lower than those of the patients (p < 0.01). Also, through k-fold cross validation, the correlation coefficient and mean average error between the CTTs measured by a conventional radiograph and the values estimated by our algorithm were 0.89 and 10.6 hours, respectively. CONCLUSIONS: The jitter and shimmer of the BS signals generated during the peristalsis could be clinically useful for the discriminative parameters of bowel motility. Also, the devised algorithm showed good potential for the continuous monitoring and estimation of bowel motility, instead of conventional radiography, and thus, it could be used as a complementary tool for the non-invasive measurement of bowel motility.

Estimation algorithm of the bowel motility based on regression analysis of the jitter and shimmer of bowel sounds.

Bowel sound (BS) signals can be used clinically as useful indicators of bowel motility. In this study, we devised a modified iterative kurtosis-based detector algorithm, in order to enhance the de-noising performance of BS signals, and an estimation algorithm of bowel motility based on the regression modeling of the jitter and shimmer of BS signals obtained by auscultation. The correlation coefficient, coefficient of determination and errors between the colon transit times measured by a conventional radiograph and the corresponding values estimated by our method were 0.987, 0.974 and 3.5 ± 3.3h, respectively. These results demonstrated that our method could be used as a complementary tool for the non-invasive diagnosis and monitoring of bowel motility, instead of conventional radiography.

Axial-scanning low-coherence interferometer method for noncontact thickness measurement of biological samples.

We investigated a high-precision optical method for measuring the thickness of biological samples regardless of their transparency. The method is based on the precise measurement of optical path length difference of the end surfaces of objects, using a dual-arm axial-scanning low-coherence interferometer. This removes any consideration of the shape, thickness, or transparency of testing objects when performing the measurement. Scanning the reference simplifies the measurement setup, resulting in unambiguous measurement. Using a 1310 nm wavelength superluminescent diode, with a 65 nm bandwidth, the measurement accuracy was as high as 11.6 µm. We tested the method by measuring the thickness of both transparent samples and nontransparent soft biological tissues.

Polarization sensitive subcutaneous and muscular imaging based on common path optical coherence tomography using near infrared source.

In this paper, we describe a polarization sensitive (PS) subcutaneous and muscular imaging system based on common path optical coherence tomography (CP-OCT) using a near infrared source. The axial and lateral resolutions of the PS-OCT system are 9 and 6 µm, respectively. The main goal of this work is to build a high-resolution and minimally invasive optical imager for examining various kinds of cutaneous substructures with intrinsic or form birefringence. The internal structural information is extracted by the real-time signal analysis (Fourier Transform) of the modulated spectral intensity depending on the beam and tissue birefringence. The preliminary results using fresh beef longissimus muscle and in vivo Rattus norvegicus (rat) show that it is possible to visualize the birefringence effect of the tissue collagen fibers in the samples in order to achieve superior image contrast and sensitivity for the detection of hidden dermal structures. Compared to conventional CP-OCT, the proposed PS-OCT system provides depth-resolved images, which reflect the tissue birefringence.

Design of the computerized 3D endoscopic imaging system for delicate endoscopic surgery.

This paper describes a 3D endoscopic video system designed to improve visualization and enhance the ability of the surgeon to perform delicate endoscopic surgery. In a comparison of the polarized and conventional electric shutter-type stereo imaging systems, the former was found to be superior in terms of both accuracy and speed for suturing and for the loop pass test. Among the groups performing loop passing and suturing, there was no significant difference in the task performance between the 2D and 3D modes, however, suturing was performed 15% (p < 0.05) faster in 3D mode by both groups. The results of our experiments show that the proposed 3D endoscopic system has a sufficiently wide viewing angle and zone for multi-viewing.

Portable measurement system for the objective evaluation of the spasticity of hemiplegic patients based on the tonic stretch reflex threshold.

The clinical scales used for the evaluation of the spasticity have some drawbacks, in spite of their simplicity and ease of assessment, and their inter- and intra-rater reliability remains controversial. The aim of this study is to develop a portable system for the objective and reliable evaluation of the spasticity based on the K-means clustering of the tonic stretch reflex threshold (TSRT) and to compare the discrimination performance of the level of spasticity determined by our method with that by the conventional modified Ashworth scale (MAS). Fifteen hemiplegic patients (7 males and 8 females, age: 63.5±15.6) participated in the study. The average and standard deviation values of the TSRTs were 127.9±1.6, 121.8±1.5 and 117.9±1.3 in groups G1, G2 and G3, respectively, and there were significant differences between the TSRTs of each group (p<0.05). Also, our groups classified by the criteria of the TSRT had a strong negative correlation (r=-0.95, r(2)=0.90, p<0.05) between the level of spasticity and TSRTs. These results demonstrated that our system could be clinically more useful for the quantitative and reliable discrimination of the spasticity than the conventional MAS.

Evaluation of Elbow Stretch Reflex Using a Portable Hand-driven Isokinetic System in Normal Adults.

OBJECTIVE: To evaluate normal healthy persons without spasticity to observe normal findings of the elbow stretch reflex using a newly developed, portable, hand-driven spasticity-measuring system. METHOD: Thirty normal persons without any disease involving the central or peripheral nervous system were enrolled in this study. The portable hand-driven isokinetic system is able to measure the joint angle, angular velocity, electromyographic (EMG) signals, and torque during elbow passive extension-flexion. One set of 10 passive elbow extension and flexion movements was performed for data acquisition at each angular velocity, including 60, 90, 120, 150 and 180 degrees per second (°/sec). Electromyographic data were collected from the biceps brachii and the triceps brachii. Torque data were collected from sensors around the wrist. RESULTS: We were able to detect EMG activity and torque in all subjects by using the new portable hand-driven isokinetic system. EMG activity and torque increased with incremental increase of angular velocities. The joint angle of maximal EMG activity according to different angular velocities did not show any significant difference (116°-127° in elbow extension and 37°-66° in elbow flexion). The joint angles of maximal torque according to different angular velocities were not significantly different either. CONCLUSION: Using the portable hand-driven isokinetic system on the elbows of normal subjects, we were able to obtain expected results. By considering our normal findings of the elbow stretch reflex using this system, we propose that the various aspects of spasticity-related data can be measured successfully.

Implementation of a multi-functional ambulatory urodynamics monitoring system based on newly devised abdominal pressure measurement.

The measurement of the rectal pressure is considered to be the 'gold standard' for the assessment of the abdominal pressure. However, conventional rectal catheters can cause erroneous results and are not comfortable for the patients. To reduce these problems, we devised a non-invasive technique for the measurement of the abdominal pressure using the parametric curve fitting method, based on linear, polynomial, exponential or sine equation modeling, between the rectal pressure and electromyographic (sEMG) signals recorded simultaneously from the abdomen. The sEMG signals and rectal pressure were obtained simultaneously from 12 patients with neurogenic bladders due to spinal cord injury (age = 53.2 ± 11.9 years, BMI = 24.4 ± 2.7, ASIA classification: D). Using our algorithm, the correlation coefficient and root mean square error (RMSE) between the measured and estimated abdominal pressure obtained by the quartic polynomial modeling were 0.86 ± 0.05 and 4.70 ± 1.56, respectively. The results obtained herein suggest that the sEMG signals can be used reliably for the indirect measurement of the abdominal pressure in ambulatory urodynamics monitoring systems.