Theoretical and experimental study of attenuation in cancellous bone.

Significance: Photoacoustic (PA) technology shows great potential for bone assessment. However, the PA signals in cancellous bone are complex due to its complex composition and porous structure, making such signals challenging to apply directly in bone analysis. Aim: We introduce a photoacoustic differential attenuation spectrum (PA-DAS) method to separate the contribution of the acoustic propagation path to the PA signal from that of the source, and theoretically and experimentally investigate the propagation attenuation characteristics of cancellous bone. Approach: We modified Biot's theory by accounting for the high frequency and viscosity. In parallel with the rabbit osteoporosis model, we build an experimental PA-DAS system featuring an eccentric excitation differential detection mechanism. Moreover, we extract a PA-DAS quantization parameter-slope-to quantify the attenuation of high- and low-frequency components. Results: The results show that the porosity of cancellous bone can be evaluated by fast longitude wave attenuation at different frequencies and the PA-DAS slope of the osteoporotic group is significantly lower compared with the normal group (**p<0.01). Conclusions: Findings demonstrate that PA-DAS effectively differentiates osteoporotic bone from healthy bone, facilitating quantitative assessment of bone mineral density, and osteoporosis diagnosis.

Loss of mouse Stmn2 function causes motor neuropathy.

Amyotrophic lateral sclerosis (ALS) is characterized by motor neuron degeneration accompanied by aberrant accumulation and loss of function of the RNA-binding protein TDP43. Thus far, it remains unresolved to what extent TDP43 loss of function directly contributes to motor system dysfunction. Here, we employed gene editing to find whether the mouse ortholog of the TDP43-regulated gene STMN2 has an important function in maintaining the motor system. Both mosaic founders and homozygous loss-of-function Stmn2 mice exhibited neuromuscular junction denervation and fragmentation, resulting in muscle atrophy and impaired motor behavior, accompanied by an imbalance in neuronal microtubule dynamics in the spinal cord. The introduction of human STMN2 through BAC transgenesis was sufficient to rescue the motor phenotypes observed in Stmn2 mutant mice. Collectively, our results demonstrate that disrupting the ortholog of a single TDP43-regulated RNA is sufficient to cause substantial motor dysfunction, indicating that disruption of TDP43 function is likely a contributor to ALS.

De novo DNA methyltransferases DNMT3A and DNMT3B are essential for XIST silencing for erosion of dosage compensation in pluripotent stem cells.

Human pluripotent stem cells (hPSCs) have proven to be valuable tools for both drug discovery and the development of cell-based therapies. However, the long non-coding RNA XIST, which is essential for the establishment and maintenance of X chromosome inactivation, is repressed during culture, thereby causing erosion of dosage compensation in female hPSCs. Here, we report that the de novo DNA methyltransferases DNMT3A/3B are necessary for XIST repression in female hPSCs. We found that the deletion of both genes, but not the individual genes, inhibited XIST silencing, maintained the heterochromatin mark of H3K27me3, and did not cause global overdosage in X-linked genes. Meanwhile, DNMT3A/3B deletion after XIST repression failed to restore X chromosome inactivation. Our findings revealed that de novo DNA methyltransferases are primary factors responsible for initiating erosion of dosage compensation in female hPSCs, and XIST silencing is stably maintained in a de novo DNA-methylation-independent manner.

C9orf72 suppresses systemic and neural inflammation induced by gut bacteria.

A hexanucleotide-repeat expansion in C9ORF72 is the most common genetic variant that contributes to amyotrophic lateral sclerosis and frontotemporal dementia1,2. The C9ORF72 mutation acts through gain- and loss-of-function mechanisms to induce pathways that are implicated in neural degeneration3-9. The expansion is transcribed into a long repetitive RNA, which negatively sequesters RNA-binding proteins5 before its non-canonical translation into neural-toxic dipeptide proteins3,4. The failure of RNA polymerase to read through the mutation also reduces the abundance of the endogenous C9ORF72 gene product, which functions in endolysosomal pathways and suppresses systemic and neural inflammation6-9. Notably, the effects of the repeat expansion act with incomplete penetrance in families with a high prevalence of amyotrophic lateral sclerosis or frontotemporal dementia, indicating that either genetic or environmental factors modify the risk of disease for each individual. Identifying disease modifiers is of considerable translational interest, as it could suggest strategies to diminish the risk of developing amyotrophic lateral sclerosis or frontotemporal dementia, or to slow progression. Here we report that an environment with reduced abundance of immune-stimulating bacteria10,11 protects C9orf72-mutant mice from premature mortality and significantly ameliorates their underlying systemic inflammation and autoimmunity. Consistent with C9orf72 functioning to prevent microbiota from inducing a pathological inflammatory response, we found that reducing the microbial burden in mutant mice with broad spectrum antibiotics-as well as transplanting gut microflora from a protective environment-attenuated inflammatory phenotypes, even after their onset. Our studies provide further evidence that the microbial composition of our gut has an important role in brain health and can interact in surprising ways with well-known genetic risk factors for disorders of the nervous system.

Quantitative calibration of sound pressure in ultrasonic standing waves using the Schlieren method.

We investigated the use of the Schlieren method to calibrate the sound pressure in an ultrasonic standing-wave field. Specifically, we derived an equation to calculate the light intensity of the diffraction fringe induced by the standing-wave field. The results indicated that the sound pressure in the standing-wave field relates to the light intensity of the diffraction fringe. Simulations and experiments were conducted to verify the theoretical calculation. We demonstrated that the ratio of the light intensity of different diffraction orders relates to the sound pressure amplitude, allowing the pressure amplitude to be calibrated with the Schlieren method. Therefore, this work presents a non-intrusive calibration method that is particularly suitable for calibrating high-frequency ultrasonic standing-wave fields.

Three-dimensional reconstruction of nonplanar ultrasound fields using Radon transform and the schlieren imaging method.

This paper introduces a numerical method of reconstructing three-dimensional (3D) ultrasound fields from their two-dimensional back-projections to understand the intensity distribution of nonplanar waves. The horizontal planes for line-focused ultrasound fields were roughly elliptical, whereas those for point-focused ultrasound fields were nearly circular. Experimental and simulated results indicated that the ultrasound intensity at the central axis can be calibrated from the light intensity by 3D reconstruction of the ultrasound field. The difference between the measured ultrasound intensity for nonplanar waves in schlieren imaging and the simulation results were discussed. From this work, the 3D ultrasound field becomes possible to be reconstructed.

A flat acoustic lens to generate a Bessel-like beam.

We report a flat acoustic lens with a periodical structure to transform a divergent beam into a Bessel-like beam. Using the Schlieren imaging technique, the propagation process of acoustic wave in the periodical structure was experimentally observed. The pressure distribution in the main lobe is in good agreement with the Bessel function and the positions of the side lobes are close to the peak of the Bessel function. To observe the directivity of the beam, simulations were performed using the finite-element method. The simulation results indicated that the transmitted acoustic intensity at the central axis was several times greater with the lens than without it. The applicability of the lens for detecting the location of an acoustic source was also investigated.

Quantitative measurement of acoustic pressure in the focal zone of acoustic lens-line focusing using the Schlieren method.

This paper proposes a theory and method for quantitative measurement of the acoustic lens-line focusing ultrasonic (ALLFU) field in its focal spot size and acoustic pressure using the Schlieren imaging technique. Using Fourier transformation, the relationship between the brightness of the Schlieren image and the acoustic pressure was introduced. The ALLFU field was simulated using finite element method and compared with the Schlieren acoustic field image. The measurement of the focal spot size was performed using the Schlieren method. The acoustic pressure in the focal zone of the ALLFU field and the transducer-transmitting voltage response were quantitatively determined by measuring the diffraction light fringe intensity. The results show that the brightness of the Schlieren image is a linear function of the acoustic intensity when the acousto-optic interaction length remains constant and the acoustic field is weak.

Assembly and molecular order of two-dimensional peptoid nanosheets through the oil-water interface.

Peptoid nanosheets are a recently discovered class of 2D nanomaterial that form from the self-assembly of a sequence-specific peptoid polymer at an air-water interface. Nanosheet formation occurs first through the assembly of a peptoid monolayer and subsequent compression into a bilayer structure. These bilayer materials span hundreds of micrometers in lateral dimensions and have the potential to be used in a variety of applications, such as in molecular sensors, artificial membranes, and as catalysts. This paper reports that the oil-water interface provides another opportunity for growth of these unique and highly ordered peptoid sheets. The monolayers formed at this interface are found through surface spectroscopic measurements to be highly ordered and electrostatic interactions between the charged moieties, namely carboxylate and ammonium residues, of the peptoid are essential in the ability of these peptoids to form ordered nanosheets at the oil-water interface. Expanding the mechanism of peptoid nanosheet formation to the oil-water interface and understanding the crucial role of electrostatic interactions between peptoid residues in nanosheet formation is essential for increasing the complexity and functionality of these nanomaterials.

Imaging and analyzing the elasticity of vascular smooth muscle cells by atomic force acoustic microscope.

Vascular smooth muscle cells (VSMCs) play an important role in the good performance of the vasculature. To study the surface, intracellular structure and elasticity of VSMCs, atomic force acoustic microscope (AFAM) was used for imaging VSMCs from A7r5 rat aorta arteries. The topography images of VSMCs were obtained in contact mode and the acoustic images were obtained by AFAM in sample vibration mode. Then, the force curve measurement derived using Young's modulus of the interested areas was used for evaluating elasticity properties. The acoustic images were found in higher resolution with more information than the topography images. The force curves showed the difference in Young's modulus of the different parts of VSMC. These findings demonstrate that AFAM is useful for displaying the surface, structure and elasticity property of VSMCs clearly, with short scanning time, negligible harm or damage to cell and nanometer-level resolution.

Cellular damage and apoptosis along with changes in NF-kappa B expression were induced with contrast agent enhanced ultrasound in gastric cancer cells and hepatoma cells.

BACKGROUND: The effect of cell injury and apoptosis induced by ultrasound with contrast agent has been verified. Contrast agent enhanced apoptosis and expression of genes that related to apoptosis and are responsive to ultrasound. This effect was associated with reactive oxygen species (ROS) production induced by the sonochemical reaction, as reported in previous studies. NF-kappa B may be one of the factors involved in oxidizing reactions or modulation during the process of ultrasound inducing apoptosis. RESULTS: Ultrasound irradiated gastric cancer cells (SGC7901 cell line) and hepatocellular carcinoma cells (SMMC-771 cell line) cultured in medium containing contrast agent. Significant cellular damage and apoptosis were observed in the bath cells incubated for 24 hours following 120 seconds ultrasonic irradiation. I kappa B alfa expression synchronously increased in the treatment groups of both the cell lines, and the down-regulated expression of NF-kappa B influenced its-regulated expression of genes that related to apoptosis. Production of intracellular ROS and elevation of NF-kappa B level occurred after incubation of the cells for 1 hour following ultrasonic treatment. CONCLUSIONS: Our result suggested that contrast agent enhanced the biological effect of ultrasound. Their reaction might stimulate the transitory expression of NF-kappaB, and subsequent elevation in IκBalfa expression could lead to the apoptosis of SGC7901 cells and SMMC-771 cells.

Acoustic field excited by a pulsed laser line source in a cylinder.

The excitation and propagation of the acoustic waves in an elastic cylinder are studied by laser ultrasonics both theoretically and experimentally. The theoretical analysis of the two-dimensional acoustic field excited by a pulsed laser line source impacting on the generatrix of an elastic cylinder is presented. The dispersive properties for both cylindrical Rayleigh wave and the higher modes--whispering gallery (WG) modes are analyzed in detail. The numerical transient displacement waveforms for a detecting point located another terminal of the cylinder diameter opposite the source are calculated. The experimental excitation and detection of the acoustic waves in an aluminum cylinder are carried out on a laser ultrasonic system, which mainly consists of a Q-switched Nd:YAG laser and a laser interferometer. The wave components of bulk waves and surface waves (cylindrical Rayleigh waves and WG modes) are analyzed by comparing the numerical and experimental waveforms. The results are in good agreement.