Mediators of mechanotransduction between bone cells

Mechanical forces are known to regulate the function of tissues in the body, including bone. Bone adapts to its mechanical environment by altering its shape and increasing its size in response to increases in mechanical load associated with exercise, and by decreasing its size in response to decreases in mechanical load associated with microgravity or prolonged bed rest. Changes in bone size and shape are produced by a cooperative action of two main types of the bone cells - osteoclasts that destroy bone and osteoblasts that build bone. These cell types come from different developmental origins, and vary greatly in their characteristics, such as size, shape, and expression of receptor subtypes, which potentially may affect their responses to mechanical stimuli. The objective of this study is to compare the responses of osteoclasts and osteoblasts to mechanical stimulation. This study has allowed us to conclude the following: 1. A mediator is released from a single source cell. 2. The response to the mediator changes with distance. 3. The value of the apparent diffusion coeficient increases with distance. 4. A plausible proposed mechanism is that ATP is released and degrades to ADP. 5. Future experiments are required to confim that ATP is the mediator as suggested

An integrated approach of GRA coupled with principal component analysis for multi-optimization of shielded metal arc welding (SMAW) process

Welding distortion is a critical issue as it leads to severe deterioration of structural integrity of welded work piece and dimensional precision. This study aims at studying the e ects of shielded metal arc welding (SMAW) parameters on the evolution of mechanical properties, including tensile strength, impact toughness, and hardness, along with angular distortion on a welded joint from SA 516 grade 70. Such parameters are analyzed and optimized by employing the Taguchi method and Grey relational analysis. SA 516 grade 70 is commercially used for fabrication of storage tanks, boilers and pressure vessels. SMAW is investigated with three levels of root gap, groove angle, electrode diameter, and pre-heat temperature, which were varied on a butt joint in flat (1 G) position to determine their e ects on response variables at room temperature. Nine experiments were designed using a Taguchi L9 orthogonal array, welded according to American Society of Mechanical Engineers (ASME) section IX, and samples were prepared and tested as per ASTM A 370. The Taguchi method and Grey relational analysis were employed to observe the most significant parameters and optimal levels that synergically yield improved responses. Results are validated by conducting confirmatory experiments that show good agreement with optimum results

Construction and evaluation of multisite recombinatorial (Gateway) cloning vectors for Gram-positive bacteria

<p>Abstract</p> <p>Background</p> <p>The Gateway recombinatorial cloning system allows easy and rapid joining of DNA fragments. Here we report the construction and evaluation of three different Gram-positive vectors that can be used with the Multisite Gateway cloning system to rapidly produce new gene arrangements in plasmid constructs for use in a variety of Gram-positive bacteria.</p> <p>Results</p> <p>Comparison of patterns of reporter gene expression with conventionally constructed clones show that the presence of residual recombination (att) sites does not have an effect on patterns of gene expression, although overall levels of gene expression may vary. Rapid construction of these new vectors allowed vector/gene combinations to be optimized following evaluation of plasmid constructs in different bacterial cells and demonstrates the benefits of plasmid construction using Gateway cloning.</p> <p>Conclusion</p> <p>The residual <it>att </it>sites present after Gateway cloning did not affect patterns of promoter induction in Gram-positive bacteria and there was no evidence of differences in mRNA stability of transcripts. However overall levels of gene expression may be reduced, possibly due to some post-transcriptional event. The new vectors described here allow faster, more efficient cloning in range of Gram-positive bacteria.</p

Blood and cardiovascular health parameters after supplementing with ketone salts for six weeks

Background: Exogenous ketone salts (KS) have been administered as treatment for various health conditions; however, the safety of chronic supplementation in a healthy population has yet to be explored. Aim: This study examined the safety of KS supplementation for 6 weeks in healthy, young adults and determined the effects of KS on blood ketone levels. Setting: Data collection occurred in a laboratory at Augusta University. Methods: Twenty-three men and women (aged 18–35 years old) supplemented with KS or a placebo (PLA) twice per day for 6 weeks in a randomised, double-blinded, PLA-controlled design. Baseline and post-intervention measures included body mass index (BMI), resting blood pressure and heart rate, questionnaires assessing mood and energy, urinalysis, and venous blood measures, including comprehensive metabolic panel (CMP), lipid panel, and complete blood count (CBC). In addition, the participants consumed the assigned supplement during the baseline and post-intervention visits; blood ketone levels were assessed immediately before and after 30 and 60 min post-supplementation. Results: Systolic blood pressure was significantly lower (p < 0.05) after supplementing with KS for 6 weeks but not PLA. All other health parameters remained unchanged by KS supplementation, including BMI, resting heart rate, urinalysis, CMP, lipid panel, and CBC. After acute administration of KS but not PLA, blood ketone levels were significantly elevated (p < 0.001) 30 and 60 min after supplementation at both baseline and post-intervention visits. Conclusion: Chronic KS supplementation seems safe, significantly elevates blood ketone levels 30 and 60 min after supplementation and may lower blood pressure. Future explorations should determine the success of KS supplementation as a strategy to combat hypertension

Reconstructing promoter activity from Lux bioluminescent reporters

The bacterial Lux system is used as a gene expression reporter. It is fast, sensitive and non-destructive, enabling high frequency measurements. Originally developed for bacterial cells, it has also been adapted for eukaryotic cells, and can be used for whole cell biosensors, or in real time with live animals without the need for euthanasia. However, correct interpretation of bioluminescent data is limited: the bioluminescence is different from gene expression because of nonlinear molecular and enzyme dynamics of the Lux system. We have developed a computational approach that, for the first time, allows users of Lux assays to infer gene transcription levels from the light output. This approach is based upon a new mathematical model for Lux activity, that includes the actions of LuxAB, LuxEC and Fre, with improved mechanisms for all reactions, as well as synthesis and turn-over of Lux proteins. The model is calibrated with new experimental data for the LuxAB and Fre reactions from Photorhabdus luminescens --- the source of modern Lux reporters --- while literature data has been used for LuxEC. Importantly, the data show clear evidence for previously unreported product inhibition for the LuxAB reaction. Model simulations show that predicted bioluminescent profiles can be very different from changes in gene expression, with transient peaks of light output, very similar to light output seen in some experimental data sets. By incorporating the calibrated model into a Bayesian inference scheme, we can reverse engineer promoter activity from the bioluminescence. We show examples where a decrease in bioluminescence would be better interpreted as a switching off of the promoter, or where an increase in bioluminescence would be better interpreted as a longer period of gene expression. This approach could benefit all users of Lux technology

Bound, virtual and resonance SS-matrix poles from the Schr\"odinger equation

A general method, which we call the potential $S$-matrix pole method, is developed for obtaining the $S$-matrix pole parameters for bound, virtual and resonant states based on numerical solutions of the Schr\"odinger equation. This method is well-known for bound states. In this work we generalize it for resonant and virtual states, although the corresponding solutions increase exponentially when $r\to\infty$. Concrete calculations are performed for the $1^+$ ground and the $0^+$ first excited states of $^{14}\rm{N}$, the resonance $^{15}\rm{F}$ states ($1/2^+$, $5/2^+$), low-lying states of $^{11}\rm{Be}$ and $^{11}\rm{N}$, and the subthreshold resonances in the proton-proton system. We also demonstrate that in the case the broad resonances their energy and width can be found from the fitting of the experimental phase shifts using the analytical expression for the elastic scattering $S$-matrix. We compare the $S$-matrix pole and the $R$-matrix for broad $s_{1/2}$ resonance in ${}^{15}{\rm F}$Comment: 14 pages, 5 figures (figures 3 and 4 consist of two figures each) and 4 table