Characterization and in vivo study of decellularized aortic scaffolds using closed sonication system.

Extracellular matrix (ECM) based bioscaffolds prepared by decellularization has increasingly emerged in tissue engineering application because it has structural, biochemical, and biomechanical cues that have dramatic effects upon cell behaviors. Therefore, we developed a closed sonication decellularization system to prepare ideal bioscaffolds with minimal adverse effects on the ECM. The decellularization was achieved at 170 kHz of ultrasound frequency in 0.1% and 2% Sodium Dodecyl Sulphate (SDS) solution for 10 hours. The immersion treatment as control was performed to compare the decellularization efficiency with our system. Cell removal and ECM structure were determined by histological staining and biochemical assay. Biomechanical properties were investigated by the indentation testing to test the stiffness, a residual force and compression of bioscaffolds. Additionally, in vivo implantation was performed in rat to investigate host tissue response. Compared to native tissues, histological staining and biochemical assay confirm the absence of cellularity with preservation of ECM structure. Moreover, sonication treatment has not affected the stiffness [N/mm] and a residual force [N] of the aortic scaffolds except for compression [%] which 2% SDS significantly decreased compared to native tissues showing higher SDS has a detrimental effect on ECM structure. Finally, minimal inflammatory response was observed after 1 and 5 weeks of implantation. This study reported that the novelty of our developed closed sonication system to prepare ideal bioscaffolds for tissue engineering applications.

Development of decellularized meniscus using closed sonication treatment system: potential scaffolds for orthopedics tissue engineering applications.

PURPOSE: Meniscus is a fibrocartilagenous tissue that cannot effectively heal due to its complex structure and presence of avascular zone. Thus, tissue engineering and regenerative medicine offer an alternative for the regeneration of meniscus tissues using bioscaffolds as a replacement for the damaged one. The aim of this study was to prepare an ideal meniscus bioscaffold with minimal adverse effect on extracellular matrix components (ECMs) using a sonication treatment system. METHODS: The decellularization was achieved using a developed closed sonication treatment system for 10 hrs, and continued with a washing process for 5 days. For the control, a simple immersion treatment was set as a benchmark to compare the decellularization efficiency. Histological and biochemical assays were conducted to investigate the cell removal and retention of the vital extracellular matrix. Surface ultrastructure of the prepared scaffolds was evaluated using scanning electron microscope at 5,000× magnification viewed from cross and longitudinal sections. In addition, the biomechanical properties were investigated through ball indentation testing to study the stiffness, residual forces and compression characteristics. Statistical significance between the samples was determined with p-value =0.05. RESULTS: Histological and biochemical assays confirmed the elimination of antigenic cellular components with the retention of the vital extracellular matrix within the sonicated scaffolds. However, there was a significant removal of sulfated glycosaminoglycans. The surface histoarchitecture portrayed the preserved collagen fibril orientation and arrangement. However, there were minor disruptions on the structure, with few empty micropores formed which represented cell lacunae. The biomechanical properties of bioscaffolds showed the retention of viscoelastic behavior of the scaffolds which mimic native tissues. After immersion treatment, those scaffolds had poor results compared to the sonicated scaffolds due to the inefficiency of the treatment. CONCLUSION: In conclusion, this study reported that the closed sonication treatment system had high capabilities to prepare ideal bioscaffolds with excellent removal of cellular components, and retained extracellular matrix and biomechanical properties.

Influence of visceral fat and blood pressure on changes in blood flow velocity in non-obese individuals.

INTRODUCTION: The aim of this study was to evaluate the impact of different visceral fat (VF) and blood pressure (BP) levels on changes in blood flow velocity (BFV) among non-obese subjects, using a cross-sectional study approach. METHODS: A total of 110 putatively healthy and non-obese subjects were divided into three groups according to their level of VF and BP. Common carotid artery BFV was measured using a developed portable Doppler ultrasound measurement system. RESULTS: The most pronounced peak systolic velocity (S1) was lower (p < 0.05) in the hypertensive group and the peak diastolic velocity (D) was significantly lower in the pre-hypertensive group than in the normotensive group. There were differences in velocity reflection and resistive indices between the hypertensive and other two BP groups. The higher VF group had significantly lower S1 and D velocities and resistive and vascular elasticity indices. By contrast, the velocity reflection index was larger in the higher VF group. CONCLUSIONS: We confirmed that there were significant differences in the BFV among non-obese subjects who differed in level of VF and BP. This study confirms that a putatively increasing VF and BP level is associated with the development of hypertension.

Role of exosomes as a proinflammatory mediator in the development of EBV-associated lymphoma.

Epstein-Barr virus (EBV) causes various diseases in the elderly, including B-cell lymphoma such as Hodgkin's lymphoma and diffuse large B-cell lymphoma. Here, we show that EBV acts in trans on noninfected macrophages in the tumor through exosome secretion and augments the development of lymphomas. In a humanized mouse model, the different formation of lymphoproliferative disease (LPD) between 2 EBV strains (Akata and B95-8) was evident. Furthermore, injection of Akata-derived exosomes affected LPD severity, possibly through the regulation of macrophage phenotype in vivo. Exosomes collected from Akata-lymphoblastoid cell lines reportedly contain EBV-derived noncoding RNAs such as BamHI fragment A rightward transcript (BART) micro-RNAs (miRNAs) and EBV-encoded RNA. We focused on the exosome-mediated delivery of BART miRNAs. In vitro, BART miRNAs could induce the immune regulatory phenotype in macrophages characterized by the gene expressions of interleukin 10, tumor necrosis factor-α, and arginase 1, suggesting the immune regulatory role of BART miRNAs. The expression level of an EBV-encoded miRNA was strongly linked to the clinical outcomes in elderly patients with diffuse large B-cell lymphoma. These results implicate BART miRNAs as 1 of the factors regulating the severity of lymphoproliferative disease and as a diagnostic marker for EBV+ B-cell lymphoma.

Imbalanced expression of polycistronic miRNA in acute myeloid leukemia.

miR-1 and miR-133 are clustered on the same chromosomal loci and are transcribed together as a single transcript that is positively regulated by ecotropic virus integration site-1 (EVI1). Previously, we described how miR-133 has anti-tumorigenic potential through repression of EVI1 expression. It has also been reported that miR-1 is oncogenic in the case of acute myeloid leukemia (AML). Here, we show that expression of miR-1 and miR-133, which have distinct functions, is differentially regulated between AML cell lines. Interestingly, the expression of miR-1 and EVI1, which binds to the promoter of the miR-1/miR-133 cluster, is correlative. The expression levels of TDP-43, an RNA-binding protein that has been reported to increase the expression, but inhibits the activity, of miR-1, were not correlated with expression levels of miR-1 in AML cells. Taken together, our observations raise the possibility that the balance of polycistronic miRNAs is regulated post-transcriptionally in a hierarchical manner possibly involving EVI1, suggesting that the deregulation of this balance may play some role in AML cells with high EVI1 expression.

Fluorescence multispectral imaging-based diagnostic system for atherosclerosis.

BACKGROUND: Composition of atherosclerotic arterial walls is rich in lipids such as cholesterol, unlike normal arterial walls. In this study, we aimed to utilize this difference to diagnose atherosclerosis via multispectral fluorescence imaging, which allows for identification of fluorescence originating from the substance in the arterial wall. METHODS: The inner surface of extracted arteries (rabbit abdominal aorta, human coronary artery) was illuminated by 405 nm excitation light and multispectral fluorescence images were obtained. Pathological examination of human coronary artery samples were carried out and thickness of arteries were calculated by measuring combined media and intima thickness. RESULTS: The fluorescence spectra in atherosclerotic sites were different from those in normal sites. Multiple regions of interest (ROI) were selected within each sample and a ratio between two fluorescence intensity differences (where each intensity difference is calculated between an identifier wavelength and a base wavelength) from each ROI was determined, allowing for discrimination of atherosclerotic sites. Fluorescence intensity and thickness of artery were found to be significantly correlated. CONCLUSIONS: These results indicate that multispectral fluorescence imaging provides qualitative and quantitative evaluations of atherosclerosis and is therefore a viable method of diagnosing the disease.

Abundance of sulfur-degrading bacteria in a benthic bacterial community of shallow sea sediment in the off-Terengganu coast of the South China Sea.

This study for the first time provides insight into the bacterial community in the benthic region of the Off-Terengganu Coastline, which is considered to be anthropogenically polluted due to heavy fishing vessel commotion. Subsurface bacteria were randomly collected from two locations at different depths and were examined using the 16S rDNA V3-V4 marker gene on the Illumina™ Miseq platform. In addition, the physiochemical parameters of the sediment were also measured. Surprisingly, the results show a high diversity of sulfur-oxidizing bacteria in the surveyed area, where Sulfurovum sp. was identified to predominate the overall bacterial community. The physiochemical parameters reveal insufficient evidence of hydrothermal vents in the surveyed area. However, there are traces of hydrocarbon pollutants such as gasoline, diesel, and mineral oil in this area. It is assumed that sediment accumulation in the lee of breakwater plays an important role in trapping the runoff from the nearby harbor, which includes oil spills. Based on the common knowledge, Sulvurofum sp. is a native bacterium that exists in deep hydrothermal vents and volcanic territories. Although the reason for the abundance of Sulfurovum sp. in the surveyed area is still unclear, there is a possibility that metabolic adaptation plays an important role in regulating hydrocarbon pollutants for survival. The work presented in this paper therefore has profound implications for future studies on Sulfurovum sp. versatility. However, future research is needed to strengthen the findings of this study and to provide a better evidence regarding the metabolic response of this bacterium toward hydrocarbon pollutants.

Wireless blood velocity spectra measurement system for healthcare evaluation: reference data.

The aim of the present study was to determine the blood velocity and its indices in common carotid artery (CCA) as reference value for clinical and healthcare application using the constructed portable wireless Doppler blood flow velocimeter system. Evaluation of CCA blood velocity waveforms was performed in 202 healthy volunteers in the age range of 20 to 69 years. Multivariate analysis was used to determine the relationship between the fixed factor and the outcome hemodynamic variables after adjustment for the related covariates. Results show a general age-related decrease in flow velocities and change in the velocity waveform. There is also a gender difference in velocity indices, except for peak systolic blood velocity. Gender difference is also found in systolic and pulse blood pressures. The findings suggest that waveform indices provide a more reliable evaluation of effects of aging and gender on CCA flow. In conclusion, normal CCA blood velocity parameters are determined in a total of 202 healthy volunteers between the third and seventh age decade after adjustment for gender and exercise effects. Findings may contribute to improved means of healthcare monitoring and clinical evaluation.

Effect of gender on blood flow velocities and blood pressure: role of body weight and height.

Aging and gender alter arterial hemodynamic function. Gender-related differences in body size may affect the arterial hemodynamics such as systolic blood pressure, wave reflection and pressure wave propagation. To assess the possibility that the differences of carotid blood velocities and brachial blood pressure in women are related to body size, we investigated the arterial hemodynamics in 50 healthy young adults (30 men, 20 women) between the age ranges of 20 to 29 years old. Blood velocities are characterized to 5 components of velocity waveforms as peak systolic (S1), second systolic (S2), incisura between systole and diastole (I), peak diastolic (D) and end-diastolic velocity wave (d). As the most pronounced systolic blood pressure is lower in young women than men (P<0.05). Women have higher diastolic velocity components and second systolic velocity, but have lower systolic velocity compared to men. From that, women have a significant greater wave reflection, smaller vascular resistive and lower vascular elastic recoil than in men. Greater body weight is associated with higher blood pressures (P<0.05) and lower blood velocities (P<0.05). Shorter body height affects not only the increased velocity wave reflection index (r= -0.519, P>0.0001), but also the decrease of peripheral vascular resistive index (RI) and arterial vascular elastic recoil index (r= 0.463, P<0.0001; r= 0.481, P<0.0001, respectively). In conclusion, body size influences blood pressure and velocity regulations in the young population.

Effects of aging and exercise training on the common carotid blood velocities in healthy men.

An age-related alteration in the cardiovascular response to exercise training are evident. The purpose of the study was to investigate the effects of exercise and age on blood velocities in common carotid artery in 82 healthy men between the age ranges of 21 to 67 years old. Blood velocities are characterized to five components of velocity waveforms as peak systolic (S1), second systolic (S2), incisura between systole and diastole (I), peak diastolic (D) and end-diastolic velocity wave (d). Decrease of blood velocities in peak systolic (r= -0.711, P<0.0001) and in peak diastolic velocities (r= -0.521, P<0.0001) with aging are improved and partially restore in particularly older men. The velocity ratio of S2/S2-1 as a reflection index increase with age (r= 0.797, P<0.0001), however is smaller in exercise-trained older compared with sedentary peers. The ratio of 1-I/D as a vascular elastic recoil index decrease with aging (r= -0.640, P<.0001), but is relatively higher in exercise-trained men. Exercise training improves the age-related deterioration in blood velocities and its indices in healthy men. In the further investigations, the assessment of aerobic fitness and vascular aging has potential by using the criteria of peak systolic and peak diastolic, and its indices.

Monitoring Carotid Blood Flow and ECG for Cardiovascular Disease in Elder Subjects.

This report is to investigate the correlation between common carotid blood flow (CCBF) properties and the risk factors for cardiovascular disease. The measurement system was designed for simultaneously monitoring of CCBF and electrocardiogram (ECG) in order to get more valid information for detection and diagnosis of cardiovascular disease. In our study, blood flow velocimeter was developed to measure blood flow in carotid by using ultrasound Doppler technique. This measurement system is based on a continuous-wave (CW) Doppler ultrasound method with two semicircular piezoelectric (PZT) transducers, one continuously transmitting ultrasound, and the other continuously receiving the echoes. Eleven patients with cardiovascular disease underwent in the experiment. In the report, blood flow was compared with data from 25 healthy subjects which asymptomatic subjects putatively free of cardiovascular disease as controls. The maximum of CCBF velocity was 102.8 (SD 18.3) cm/s in the 25 healthy subjects. While, the maximum of CCBF velocities were 53.9 (SD 16.6) cm/s in the eleven cardiac patients. The data suggest that CCBF velocity decreased significantly in the cardiac patients and by simultaneously monitoring of CCBF and ECG were probably obtained more valid information to detect and diagnose cardiovascular disease at the early stage.

A change of blood flow during strenuous physical exercises using cycle ergometer.

Our purpose of the study is to measure the blood flow in the carotid artery during physical exercises. We have developed the blood flow velocimeter during physical exercises by using the non-invasive Doppler ultrasound method. We used it and measured blood flow in the carotid artery in which using a cycle ergometer and had two exercise tests in this report. The protocol exercise with transient response and ramp response were conducted. As a result, maximum blood flow velocities changed largely during physical exercises. In particular the maximum blood flow velocity was decreased during physical exercise in transient response. In addition, we could also detect arrhythmia that occurred in the subject immediately after exercise. For this reason, blood flow measurement during physical exercise maybe useful for the early detection of cardiovascular disease. Recently, the physical exercises are very popular for health maintenance.