Validation of housekeeping genes as internal controls for gene expression studies on biofilm formation in Bacillus velezensis.

Bacillus velezensis is an important bacterium widely applied in agriculture and industry, and biofilms play critical roles in its environmental tolerance. The appropriate choice of reference genes is essential for key gene expression studies. Multiple internal control genes were selected and validated from the 21 housekeeping genes of B. velezensis by expression stability evaluation during biofilm formation and were used to study the expression of key genes involved in the process. The results showed that pyk, gyrA, recA, and gyrB were stably expressed, and the expression of pyk was the most stable during biofilm formation. A pair of two genes, pyk and gyrA, provided high-quality data when used as internal controls, and the combination of three genes, pyk, gyrA, and recA, was even better. The expression levels of pyk, gyrA, and recA approximated those of five key genes, abrB, epsD, kinC, sinR, and tasA, in biofilm formation, meeting the requirements of ideal internal control genes. The expression patterns of 5 key genes were studied with 16S, pyk, the pair of 2 genes, pyk and gyrA, and the combination of 3 genes, pyk, gyrA, and recA, as internal controls during the biofilm formation process. The results proved that pyk was a suitable internal control, as were the pair of 2 genes, pyk and gyrA, and the combination of 3 genes, pyk, gyrA, and recA. This study provided genes and gene combinations which were validated as suitable internal controls for gene expression studies, especially those on the mechanism of biofilm formation in B. velezensis or even other Bacillus spp. KEY POINTS: • Reference genes is necessary for gene expression study in biofilm formation of Bacillus velezensis • Pyk and 2 gene combinations were selected and validated from 21 common used genes • Expression of key genes in biofilm formation was normalized with the selected internal controls.

Modeling Focused-Ultrasound Response for Non-Invasive Treatment Using Machine Learning.

The interactions between body tissues and a focused ultrasound beam can be evaluated using various numerical models. Among these, the Rayleigh-Sommerfeld and angular spectrum methods are considered to be the most effective in terms of accuracy. However, they are computationally expensive, which is one of the underlying issues of most computational models. Typically, evaluations using these models require a significant amount of time (hours to days) if realistic scenarios such as tissue inhomogeneity or non-linearity are considered. This study aims to address this issue by developing a rapid estimation model for ultrasound therapy using a machine learning algorithm. Several machine learning models were trained on a very-large dataset (19,227 simulations), and the performance of these models were evaluated with metrics such as Root Mean Squared Error (RMSE), R-squared (R2), Akaike Information Criterion (AIC), and Bayesian Information Criterion (BIC). The resulted random forest provides superior accuracy with an R2 value of 0.997, an RMSE of 0.0123, an AIC of -82.56, and a BIC of -81.65 on an external test dataset. The results indicate the efficacy of the random forest-based model for the focused ultrasound response, and practical adoption of this approach will improve the therapeutic planning process by minimizing simulation time.

Design and characterization of a cough simulator.

Expiratory droplets from human coughing have always been considered as potential carriers of pathogens, responsible for respiratory infectious disease transmission. To study the transmission of disease by human coughing, a transient repeatable cough simulator has been designed and built. Cough droplets are generated by different mechanisms, such as the breaking of mucus, condensation and high-speed atomization from different depths of the respiratory tract. These mechanisms in coughing produce droplets of different sizes, represented by a bimodal distribution of 'fine' and 'coarse' droplets. A cough simulator is hence designed to generate transient sprays with such bimodal characteristics. It consists of a pressurized gas tank, a nebulizer and an ejector, connected in series, which are controlled by computerized solenoid valves. The bimodal droplet size distribution is characterized for the coarse droplets and fine droplets, by fibrous collection and laser diffraction, respectively. The measured size distributions of coarse and fine droplets are reasonably represented by the Rosin-Rammler and log-normal distributions in probability density function, which leads to a bimodal distribution. To assess the hydrodynamic consequences of coughing including droplet vaporization and polydispersion, a Lagrangian model of droplet trajectories is established, with its ambient flow field predetermined from a computational fluid dynamics simulation.

Deformations experienced in the human skin, adipose tissue, and fascia in osteopathic manipulative medicine.

CONTEXT: Osteopathic manipulative medicine techniques involve compressive and tangential forces to target the fascia. These forces are transmitted to the skin and adipose tissue before the fascia is encountered. Knowing the extent of deformation of these 2 tissue layers relative to the fascia will assist osteopathic physicians in evaluating techniques for manual therapies and adjusting these therapies to reduce patient discomfort and improve results. OBJECTIVE: To determine the magnitude of the forces transmitted to the skin, adipose tissue, and fascia, and to determine the magnitude of deformation produced in the skin and adipose tissue relative to the fascia using a mathematical model. METHODS: The large deformation theory of elasticity, valid for 3-dimensional deformations, was used to evaluate the forces that need to be applied such that a specified deformation is produced in any region of the skin, adipose tissue, or fascia layers. Similarly, if the forces are specified, then the deformation produced can be determined. RESULTS: The normal and tangential forces required to produce a deformation of 9% compression and 4% shear for the skin were 50 N and 11 N, respectively. Normal and tangential forces of about 100 N and 22 N were found for a similar deformation of fascia. For adipose tissue, these forces were 36 N and 8 N, respectively. In addition, the skin experienced more compression and shear-about 1.5 times as much as the fascia, and the adipose tissue experienced about 2.5 to 3.5 times the deformation of the fascia and 50% more than the skin when a given force was applied to the skin. CONCLUSION: The forces applied to the surface of the skin were transmitted through this layer and the adipose layer entirely to the fascia. Therefore, the skin and adipose tissue experienced the same magnitude of force as the fascia. However, the skin and adipose tissue experienced more compression and shear than the fascia.

Measurement of balance in computer posturography: Comparison of methods--A brief review.

Some symptoms related to disequilibrium may not be detected by a clinical exam. Therefore, objective study is important in assessing balance. In this paper, methods to measure balance in computer posturography are compared. Center of pressure (COP) displacement, equilibrium score (ES) and postural stability index (PSI), the main measures of assessing balance are described and their merits and disadvantages are discussed. Clinicians should apply that measure which suits the specific strategies in a specific situation. Measuring devices such as Force plate, Balance Master and Equitest are also discussed. Although the Balance Master and Equitest devices are more costly compared to the force plate only, they are more useful for assessing balance relevant to daily life activities that might result in falls.

Viscoelastic stresses on anisotropic annulus fibrosus of lumbar disk under compression, rotation and flexion in manual treatment.

BACKGROUND: Twisting (spinal rotation) and bending (flexion) are commonly reported as triggers for low back pain. This paper addresses whether the twisting stress on the annulus fibrosus of the lumbar disk is greater or less than the bending stress for the same angle of twist or bending. METHODS: Stress-strain relation for transversely isotropic material is applied to the transversely isotropic annulus fibrosus of the lumbar disk to analyze the viscoelastic stresses produced due to 6% compression, 10 degrees twist and 10 degrees bending. FINDINGS: The bending stress is 450 times greater than the twisting stress for the same angle of twist or bending of the annulus fibrosus. The twisting and bending moments increase two-fold in quick maneuvers lasting 0.1s (as in high velocity manipulations), compared to slow maneuvers lasting 60s. INTERPRETATION: From biomechanical perspective, in situations where both flexion and spinal rotation occur, the stress on the intervertebral disk is markedly higher with flexion compared to rotation. In patients with low back pain that has a disk mediated (discogenic) component, manipulation and mobilization therapies should avoid flexion to minimize stress on the disks. This is particularly relevant for high velocity manipulations where the stress on the disk is doubled for both flexion and rotation. The results in this paper can help guide manual therapists to adjust their treatments to minimize stress on the intervertebral disk.

Three-dimensional mathematical model for deformation of human fasciae in manual therapy.

CONTEXT: Although mathematical models have been developed for the bony movement occurring during chiropractic manipulation, such models are not available for soft tissue motion. OBJECTIVE: To develop a three-dimensional mathematical model for exploring the relationship between mechanical forces and deformation of human fasciae in manual therapy using a finite deformation theory. METHODS: The predicted stresses required to produce plastic deformation were evaluated for a volunteer subject's fascia lata, plantar fascia, and superficial nasal fascia. These stresses were then compared with previous experimental findings for plastic deformation in dense connective tissues. Using the three-dimensional mathematical model, the authors determined the changing amounts of compression and shear produced in fascial tissue during 20 seconds of manual therapy. RESULTS: The three-dimensional model's equations revealed that very large forces, outside the normal physiologic range, are required to produce even 1% compression and 1% shear in fascia lata and plantar fascia. Such large forces are not required to produce substantial compression and shear in superficial nasal fascia, however. CONCLUSION: The palpable sensations of tissue release that are often reported by osteopathic physicians and other manual therapists cannot be due to deformations produced in the firm tissues of plantar fascia and fascia lata. However, palpable tissue release could result from deformation in softer tissues, such as superficial nasal fascia.

Square wave anodic stripping voltammetric determination of Pb2+ using acetylene black paste electrode based on the inducing adsorption ability of I-.

Herein, a sensitive and simplified electrochemical method was proposed for the determination of trace levels of Pb2+ by anodic stripping voltammetry (ASV) based on the inducing adsorption ability of I- toward Pb2+. In the presence of low concentration of I-, Pb2+ was induced to accumulate onto the acetylene black (AB) paste electrode surface, and then reduced at -0.90 V. During the following square wave sweep from -0.90 to -0.30 V, the reduced Pb was oxidized, resulting in a sensitive and well-shaped stripping peak at -0.56 V. Further studies indicate that low concentration of I- significantly enhances the sensitivity of determination of Pb2+. After all the experimental parameters were optimized, a novel and sensitive method was developed for the electrochemical determination of Pb2+. The linear range is found to be from 2.0 x 10(-8) to 4.0 x 10(-6) mol L(-1), and the lowest detectable concentration is estimated to be 6.0 x 10(-9)mol L(-1). This newly proposed method was finally demonstrated with water samples. Otherwise, the anodic stripping responses of Pb2+ on AB paste electrode and graphite paste electrode were compared.

Postural stability index is a more valid measure of stability than equilibrium score.

Researchers, therapists, and physicians often use equilibrium score (ES) from the Sensory Organization Test, a key test in the NeuroCom EquiTest System (a dynamic posturography system) to assess stability. ES reflects the overall coordination of the visual, proprioceptive, and vestibular systems for maintaining standing posture. In our earlier article, we proposed a new measure of anterior-posterior (A-P) postural stability called the Postural Stability Index (PSI), which accounts for more biomechanical aspects than ES. This article showed that PSI provides a clinically important adjunct to ES. In the present article, we show that PSI can provide an acceptable index even if a person falls during the trial, whereas ES assigns a zero score for any fall. We also show that PSI decreases as ankle stiffness increases, which is intuitive, while ES exhibits the opposite behavior. Ankle stiffness is generally recognized as an indicator of postural stability. These results suggest that PSI is a more valid measure of A-P stability than ES.

Computational method to evaluate ankle postural stiffness with ground reaction forces.

We examined an existing method for evaluating postural sway based on force-plate technology. Through an improved mathematical model of postural dynamics, we propose a new method, which better evaluated postural sway and, in addition, computed ankle moment and ankle postural stiffness directly from the measured ground reaction forces. An example is detailed that demonstrates the utility of this approach. The proposed method does not involve filtering or numerical integration and considers the platform inclination. Results from normal subjects show a linear relation between the ankle moment and the sway angle during quiet standing.

Measures of postural stability.

Dynamic posturography has become an important tool for understanding standing balance in clinical settings. A key test in the NeuroCom International (Clackamas, Oregon) dynamic posturography system, the Sensory Organization Test (SOT), provides information about the integration of multiple components of balance. The SOT test leads to an outcome measure called the "equilibrium score" (ES), which reflects the overall coordination of the visual, proprioceptive, and vestibular systems for maintaining standing posture. Researchers, therapists, and physicians often use the ES from the SOT as a clinically relevant measure of standing balance. We discuss here the formula used for evaluating the ES and propose an additional measure of postural stability, called the Postural Stability Index (PSI), that accounts for shear force and individual anthropomorphic measures. We propose that this new measure provides a clinically important adjunct to the current SOT and can be calculated from data already collected by the NeuroCom forceplate during the SOT.