Similar Biomechanical Behavior in Gait Analysis between Ceramic-on-Ceramic and Ceramic-on-XLPE Total Hip Arthroplasties

In vitro measurements are widely used to implement gait kinematic and kinetic parameters to predict THA wear rate. Clinical tests of materials and designs are crucial to prove the accuracy and validate such measurements. This research aimed to examine the effect of CoC and CoXLPE kinematics and kinetics on wear during gait, the essential functional activity of humans, by comparing in vivo data to in vitro results. Our study hypothesis was that both implants would present the same hip joint kinematics and kinetics during gait. In total, 127 unilateral primary cementless total hip arthroplasties were included in the research. There were no statistically significant differences observed at mean peak abduction, flexion, and extension moments and THA kinematics between the two groups. THA gait kinematics and kinetics are crucial biomechanical inputs associated with implant wear. In vitro studies report less wear in CoC than CoXLPE when tested in a matched gait kinematic protocol. Our findings confirm that both implants behave identically in terms of kinematics in a clinical environment, thus strengthening CoC advantage in in vitro results. Correlated to all other significant factors that affect THA wear, it could address in a complete prism the wear on CoC and CoXLPE.

Ultrasound Thermometry for HIFU-Therapy

High-Intensity Focused Ultrasound (HIFU) is an alternative tumour therapy with the ability for non-invasive thermal ablation of tissue. For a safe application, the heat deposition needs to be monitored over time, which is currently done with Magnetic Resonance Imaging. Ultrasound (US) based monitoring is a promising alternative, as it is less expensive and allows the use of a single device for both therapy and monitoring. In this work, a method for spatial and temporal US thermometry has been investigated based on simulation studies and in-vitro measurements. The chosen approach is based on the approximately linear dependence between temperature and speed of sound (SoS) in tissue for a given temperature range. By tracking the speckles of successive B-images, the possibility of detecting local changes in SoS and therefore in temperature is given. A speckle tracking algorithm was implemented for 2D and 3D US thermometry using a spatial compounding method to reduce artifacts. The algorithm was experimentally validated in an agar-based phantom and in porcine tissue for temperature rises up to △ 8°C. We used a focusing single element US transducer as therapeutic probe, a linear (/matrix array) transducer with 128 (/32∙32) elements for imaging and thermocouples for validation and calibration. In all experiments, both computational and in-vitro, we succeeded in monitoring the thermal induced SoS changes over time. The in-vitro measurements were in good agreement with the simulation results and the thermocouple measurements (rms temperature difference = 0.53 °C, rms correlation coefficient = 0. 96).

Interrogating RNA-small molecule interactions with structure probing and AI augmented-molecular simulations

While there is increasing interest in the study of RNA as a therapeutic target, efforts to understand RNA-ligand recognition at the molecular level lag far behind our understanding of protein-ligand recognition. This problem is complicated due to the more than ten orders of magnitude in timescales involved in RNA dynamics and ligand binding events, making it not straightforward to design experiments or simulations. Here we make use of artificial intelligence (AI)-augmented molecular dynamics simulations to directly observe ligand dissociation for cognate and synthetic ligands from a riboswitch system. The site-specific flexibility profiles from our simulations are in excellent agreement with in vitro measurements of flexibility using Selective 2' Hydroxyl Acylation analyzed by Primer Extension and Mutational Profiling (SHAPE-MaP). Our simulations reproduce known binding affinity profiles for the cognate and synthetic ligands, and pinpoint how both ligands make use of different aspects of riboswitch flexibility. On the basis of our dissociation trajectories, we also make and validate predictions of pairs of mutations for both the ligand systems that would show differing binding affinities. These mutations are distal to the binding site and could not have been predicted solely on the basis of structure. The methodology demonstrated here shows how molecular dynamics simulations with all-atom force-fields have now come of age in making predictions that complement existing experimental techniques and illuminate aspects of systems otherwise not trivial to understand.

Engineered bacteria detect tumor DNA in vivo

In vitro nucleic acid analysis has become a valuable diagnostic tool. However, in vitro measurements have many disadvantages when compared to in vivo techniques. Synthetic bacterial biosensors have been engineered to sense many target signals in vivo, but no biosensor exists to detect specific DNA sequences. Here, we engineered naturally competent Acinetobacter baylyi bacteria to detect engineered donor DNA inserted into the genomes of colorectal cancer (CRC) cells and organoids. The DNA biosensor concept was developed in vitro and then validated in vivo with sensor bacteria delivered orally or rectally to mice that had been injected with orthotopic donor CRC organoids. Horizontal gene transfer occurred from the donor tumor to the sensor bacteria in vivo, conferring antibiotic resistance to the sensor bacteria and allowing their detection in stool. The sensor bacteria differentiated mice with and without CRC. Life detecting life has many implications for future diagnosis, prevention, and treatment of disease. This approach may also be useful in any application that requires the detection of mutations or organisms within environments that are difficult to sample.

Evaluation of the Binding Kinetics of RHEB with mTORC1 by In-Cell and In Vitro Assays

The mammalian/mechanistic target of rapamycin complex 1 (mTORC1) is activated by the small G-protein, Ras homolog enriched in brain (RHEB–GTPase). On lysosome, RHEB activates mTORC1 by binding the domains of N-heat, M-heat, and the focal adhesion targeting (FAT) domain, which allosterically regulates ATP binding in the active site for further phosphorylation. The crucial role of RHEB in regulating growth and survival through mTORC1 makes it a targetable site for anti-cancer therapeutics. However, the binding kinetics of RHEB to mTORC1 is still unknown at the molecular level. Therefore, we studied the kinetics by in vitro and in-cell protein–protein interaction (PPI) assays. To this end, we used the split-luciferase system (NanoBiT®) for in-cell studies and prepared proteins for the in vitro measurements. Consequently, we demonstrated that RHEB binds to the whole mTOR both in the presence or absence of GTPγS, with five-fold weaker affinity in the presence of GTPγS. In addition, RHEB bound to the truncated mTOR fragments of N-heat domain (∆N, aa 60–167) or M-heat domain (∆M, aa 967–1023) with the same affinity in the absence of GTP. The reconstructed binding site of RHEB, ∆N-FAT-M, however, bound to RHEB with the same affinity as ∆N-M, indicating that the FAT domain (∆FAT, aa 1240–1360) is dispensable for RHEB binding. Furthermore, RHEB bound to the truncated kinase domain (∆ATP, aa 2148–2300) with higher affinity than to ∆N-FAT-M. In conclusion, RHEB engages two different binding sites of mTOR, ∆N-FAT-M and ∆ATP, with higher affinity for ∆ATP, which likely regulates the kinase activity of mTOR through multiple different biding modes.

Preparation, Characterization and Evaluation of Organogel-Based Lipstick Formulations: Application in Cosmetics

1,3:2,4-Dibenzylidene-D-sorbitol (DBS) and 12-hydroxystearic acid (12-HSA) are well-known as low-molecular-weight organogelators (LMOGs) capable of gelling an organic liquid phase. Considering their unique chemical and physical properties, we assessed their potential effects in new lipstick formulations by discrimination testing; in vitro measurements of the sun protection factor (SPF); and thermal, mechanical and texture analyzes. DBS and 12-HSA were used to formulate four types of lipsticks: L1 (1% DBS), L2 (10% 12-HSA), L3 (1.5% DBS) and L4 (control, no LMOGs). The lipsticks were tested for sensory perception with an untrained panel of 16 consumers. LMOG formulations exhibited higher UVA protection factor (UVA-PF) and in vitro SPF, particularly in the 12-HSA-based lipstick. Regarding thermal properties, the 12-HSA-based lipstick and those without LMOGs were more heat-amenable compared to thermoresistant DBS-based lipsticks. The results also showed the viscoelastic and thermally reversible properties of LMOGs and their effect of increasing pay-off values. In general, the texture analysis indicated that 12-HSA-based lipstick was significantly harder to bend compared to control, while the other formulations became softer and easier to bend throughout the stability study. This work suggests the potential use of LMOGs as a structuring agent for lipsticks, paving the way towards more photoprotective and sustainable alternatives.

Textile Antenna-Sensor for In Vitro Diagnostics of Diabetes

In this paper, a feasibility study of a microwave antenna-based sensor is proposed for in vitro experiments for monitoring blood glucose levels. The proposed device consists of a square-ring incorporated within a fully textile monopole antenna to absorb and sense different glucose concentrations, covering patients with different diabetic conditions. The designed antenna-sensor is optimized to operate at 2.4 GHz. The sensing principle is based on the resonance frequency shift of the reflection response of the antenna-based sensor under different glucose levels. The experiments were carried out with blood mimicking by means of aqueous solutions, using D(+)- glucose/water in different concentrations for various diabetic conditions of type-2 diabetes. The performance of the embroidered antenna-based sensor is characterized and validated using a convenient setup for in vitro measurements. The results demonstrated the ability of the proposed antenna-based sensor to cover all the glucose levels of the diabetes range, including hypoglycemia (10–70 mg/dL), normoglycemia (80–110 mg/dL) and hyperglycemia (130–190 mg/dL) with a sensitivity of 350 kHz/(mg/dL). Besides its ability to detect different glucose concentrations of various diabetic conditions, the proposed antenna-sensor presents diverse features such as a simplistic design, compact size, wearability and low cost. The proposed textile device demonstrates a proof of concept for efficient in vitro blood glucose level measurements and diagnostics of diabetes.

Switching an active site helix in dihydrofolate reductase reveals limits to sub-domain modularity

To what degree are individual structural elements within proteins modular such that similar structures from unrelated proteins can be interchanged? We study sub-domain modularity by creating 20 chimeras of an enzyme, E. coli dihydrofolate reductase (DHFR), in which a catalytically important, 10-residue α-helical sequence is replaced by α-helical sequences from a diverse set of proteins. The chimeras stably fold but have a range of diminished thermal stabilities and catalytic activities. Evolutionary coupling analysis indicates that the residues of this α-helix are under selection pressure to maintain catalytic activity in DHFR. We performed molecular dynamics simulations using replica exchange with solute-tempering. Chimeras with low catalytic activity exhibit non-helical conformations that block the binding site and disrupt the positioning of the catalytically essential residue D27. Simulation observables and in vitro measurements of thermal stability and substrate binding affinity are strongly correlated. Several E. coli strains with chromosomally integrated chimeric DHFRs can grow, with growth rates that follow predictions from a kinetic flux model that depends on the intracellular abundance and catalytic activity of DHFR. Our findings show that although α-helices are not universally substitutable, the molecular and fitness effects of modular segments can be predicted by the biophysical compatibility of the replacement segment.

Assessment of Vehicle Volatility and Deposition Layer Thickness in Skin Penetration Models

Systemic disposition of dermally applied chemicals is often formulation-dependent. Rapid evaporation of the vehicle can result in crystallization of active compounds, limiting their degree of skin penetration. In addition, the choice of vehicle can affect the permeant’s degree of penetration into the stratum corneum. The aim of this study is to build a predictive, mechanistic, dermal absorption model that accounts for vehicle-specific effects on the kinetics of permeant transport into skin. An existing skin penetration model is extended to explicitly include the effect of vehicle volatility over time. Using in vitro measurements of skin penetration by chemicals applied in both a saline and an ethanol solvent, the model is optimized to learn two vehicle-specific quantities: the solvent evaporation rate and the extent of permeant deposition into the upper stratum corneum immediately following application. The dermal disposition estimates of the trained model are subsequently compared against those of the original model using further in vitro measurements. The trained model showed a 1.5-fold improvement and a 19-fold improvement in overall goodness of fit among compounds tested in saline and ethanol solvents, respectively. The proposed model structure can thus form a basis for in vitro to in vivo extrapolations of dermal disposition for skin formulations containing volatile components.