Clinical study of probiotics combined with lactulose for minimal hepatic encephalopathy treatment.

OBJECTIVE: This study aimed to investigate the clinical effect of probiotics combined with lactulose for minimal hepatic encephalopathy (MHE) treatment. METHODS: A total of 88 patients with MHE were randomly divided into the control ( n  = 44) and combined groups ( n  = 44). The control group was treated with lactulose, while the combined group was treated with probiotics and lactulose. Serum ammonia, liver function [alanine aminotransferase (ALT) and aspartate transaminase (AST)], intestinal mucosal barrier markers [fatty acid-binding protein 2 (FABP2) and advanced glycation end-products (AGEs)] and number connection test A (NCT-A) and digit symbol test (DST) scores were tested and compared between the two groups. RESULTS: The post-treatment in the control and combined groups shows decreased serum ammonia levels, ALT and AST levels, FABP2 and AGEs levels and NCT-A score and increased DST score compared with pre-treatment, with a significant difference ( P  < 0.05). Post-treatment, the serum ammonia level, ALT and AST levels, FABP2 and AGEs levels, NCT-A score decreased and DST score increased in the combined group compared with the control group, with a significant difference ( P  < 0.05). CONCLUSION: Probiotics can promote lactulose in MHE treatment.

Development of a biotin-streptavidin-enhanced enzyme-linked immunosorbent assay (BA-ELISA) for high-throughput screening of KRASG12C inhibitors.

KRAS, the most frequently mutated oncogene in human cancers, was considered "undruggable" until the identification of small molecules that bind irreversibly to the mutant reactive cysteine at residue 12. Despite the encouraging anticancer activity of KRASG12C inhibitors in clinical trials, identification of more potent drugs is expected to achieve the maximal clinical benefit, which is hindered by the low sensitivity or throughput of current biochemical approaches. To overcome these limitations, a biotin-streptavidin-enhanced enzyme-linked immunosorbent assay (BA-ELISA) based on the competitive interaction of biotin-labeled probe and the test compound with KRASG12C was developed. Compared with reported assays, less protein was used in BA-ELISA, which significantly improves the resolution of inhibitor potency, thus contributing to the identification of highly potent inhibitors. Furthermore, BA-ELISA can also be expanded to determine the cellular potency of the inhibitors using KRASG12C mutant living cells. Using three previously disclosed compounds, ARS-1620, AMG 510, and MRTX849, we demonstrated that BA-ELISA is a highly sensitive, specific, and robust method for high-throughput screening of KRASG12C inhibitors.

A technique for delineating the unfolding requirements for substrate entry into retrotranslocons during endoplasmic reticulum-associated degradation.

The endoplasmic reticulum-associated degradation (ERAD) pathway mediates the endoplasmic reticulum-to-cytosol retrotranslocation of defective proteins through protein complexes called retrotranslocons. Defective proteins usually have complex conformations and topologies, and it is unclear how ERAD can thread these conformationally diverse protein substrates through the retrotranslocons. Here, we investigated the substrate conformation flexibility necessary for transport via retrotranslocons on the ERAD-L, ERAD-M, and HIV-encoded protein Vpu-hijacked ERAD branches. To this end, we appended various ERAD substrates with specific domains whose conformations were tunable in flexibility or tightness by binding to appropriate ligands. With this technique, we could define the capacity of specific retrotranslocons in disentangling very tight, less tight but well-folded, and unstructured conformations. The Hrd1 complex, the retrotranslocon on the ERAD-L branch, permitted the passage of substrates with a proteinase K-resistant tight conformation, whereas the E3 ligase gp78-mediated ERAD-M allowed passage only of nearly completely disordered but not well-folded substrates and thus may have the least unfoldase activity. Vpu-mediated ERAD, containing a potential retrotranslocon, could unfold well-folded substrates for successful retrotranslocation. However, substrate retrotranslocation in Vpu-mediated ERAD was blocked by enhanced conformational tightness of the substrate. On the basis of these findings, we propose a mechanism underlying polypeptide movement through the endoplasmic reticulum membrane. We anticipate that our biochemical system paves the way for identifying the factors necessary for the retrotranslocation of membrane proteins.

A Numerical Investigation into the Effects of Overweight and Obesity on Total Knee Arthroplasty.

Overweight and obesity increase risks of knee osteoarthritis, which is a major cause of disability. Severe knee osteoarthritis can be treated by knee arthroplasty. Total knee arthroplasty has been used in overweight and obese patients; however, clinical reports showed that the outcome of this group of patients was not good as normal-weight patients. Two computer models were created in this paper to simulate the effect of excess loads on the distal femoral bone and contact pressures in total knee arthroplasty during a gait cycle. The numerical results showed increased stress in periprosthetic distal femoral bones and higher contact pressure on tibial polyethylene insert during the stance phase. Based on the computer simulation results and published research work, cementless total knee arthroplasty with thicker tibial polyethylene insert may be a better option for overweight patients.

A Numerical Investigation into the Effects of Overweight and Obesity on Total Knee Arthroplasty

Overweight and obesity increase risks of knee osteoarthritis, which is a major cause of disability. Severe knee osteoarthritis can be treated by knee arthroplasty. Total knee arthroplasty has been used in overweight and obese patients; however, clinical reports showed that the outcome of this group of patients was not good as normal-weight patients. Two computer models were created in this paper to simulate the effect of excess loads on the distal femoral bone and contact pressures in total knee arthroplasty during a gait cycle. The numerical results showed increased stress in periprosthetic distal femoral bones and higher contact pressure on tibial polyethylene insert during the stance phase. Based on the computer simulation results and published research work, cementless total knee arthroplasty with thicker tibial polyethylene insert may be a better option for overweight patients.

Exploring inter-subject anatomic variability using a population of patient-specific femurs and a statistical shape and intensity model.

This paper is motivated by the need to accurately and efficiently measure key periosteal and endosteal parameters of the femur, known to critically influence hip biomechanics following arthroplasty. The proposed approach uses statistical shape and intensity models (SSIMs) to represent the variability across a wide range of patients, in terms of femoral shape and bone density. The approach feasibility is demonstrated by using a training dataset of computer tomography scans from British subjects aged 25-106 years (75 male and 34 female). For each gender, a thousand new virtual femur geometries were generated using a subset of principal components required to capture 95% of the variance in both female and male training datasets. Significant differences were found in basic anatomic parameters between females and males: anteversion, CCD angle, femur and neck lengths, head offsets and radius, cortical thickness, densities in both Gruen and neck zones. The measured anteversion for female subjects was found to be twice as high as that for male subjects: 13 ± 6.4° vs. 6.3 ± 7.8° using the training datasets compared to 12.96 ± 6.68 vs. 5.83 ± 9.2 using the thousand virtual femurs. No significant differences were found in canal flare indexes. The proposed methodology is a valuable tool for automatically generating a large specific population of femurs, targeting specific patients, supporting implant design and femoral reconstructive surgery.

Inter-subject variability effects on the primary stability of a short cementless femoral stem.

This paper is concerned with the primary stability of the Furlong Evolution(®) cementless short stem across a spectrum of patient morphology. A computational tool is developed that automatically selects and positions the most suitable stem from an implant system made of a total of 48 collarless stems to best match a 3D model based on a library of CT femur scans (75 males and 34 females). Finite Element contact models of reconstructed hips, subjected to physiologically-based boundary constraints and peak loads of walking mode, were simulated using a coefficient of friction of 0.4 and an interference-fit of 50 µm. Maximum and average implant micromotions across the subpopulation were predicted to be 100±7 µm and 7±5 µm with ranges [15 µm, 350 µm] and [1 µm, 25 µm], respectively. The computed percentage of implant area with micromotions greater than reported critical values of 50 µm, 100 µm and 150 µm never exceeded 14%, 8% and 7%, respectively. To explore the possible correlations between anatomy and implant performance, response surface models for micromotion metrics were constructed. Detailed morphological analyses were conducted and a clear nonlinear decreasing trend was observed between implant average micromotion and both the metaphyseal canal flare indices and average densities in Gruen zones. The present study demonstrates that the primary stability and tolerance of the short stem to variability in patient anatomy were high, reducing the need for patient stratification. In addition, the developed tool could be utilised to support implant design and planning of femoral reconstructive surgery.

Developing a musculoskeletal model of the primate skull: predicting muscle activations, bite force, and joint reaction forces using multibody dynamics analysis and advanced optimisation methods.

An accurate, dynamic, functional model of the skull that can be used to predict muscle forces, bite forces, and joint reaction forces would have many uses across a broad range of disciplines. One major issue however with musculoskeletal analyses is that of muscle activation pattern indeterminacy. A very large number of possible muscle force combinations will satisfy a particular functional task. This makes predicting physiological muscle recruitment patterns difficult. Here we describe in detail the process of development of a complex multibody computer model of a primate skull (Macaca fascicularis), that aims to predict muscle recruitment patterns during biting. Using optimisation criteria based on minimisation of muscle stress we predict working to balancing side muscle force ratios, peak bite forces, and joint reaction forces during unilateral biting. Validation of such models is problematic; however we have shown comparable working to balancing muscle activity and TMJ reaction ratios during biting to those observed in vivo and that peak predicted bite forces compare well to published experimental data. To our knowledge the complexity of the musculoskeletal model is greater than any previously reported for a primate. This complexity, when compared to more simple representations provides more nuanced insights into the functioning of masticatory muscles. Thus, we have shown muscle activity to vary throughout individual muscle groups, which enables them to function optimally during specific masticatory tasks. This model will be utilised in future studies into the functioning of the masticatory apparatus.

The application of muscle wrapping to voxel-based finite element models of skeletal structures.

Finite elements analysis (FEA) is now used routinely to interpret skeletal form in terms of function in both medical and biological applications. To produce accurate predictions from FEA models, it is essential that the loading due to muscle action is applied in a physiologically reasonable manner. However, it is common for muscle forces to be represented as simple force vectors applied at a few nodes on the model's surface. It is certainly rare for any wrapping of the muscles to be considered, and yet wrapping not only alters the directions of muscle forces but also applies an additional compressive load from the muscle belly directly to the underlying bone surface. This paper presents a method of applying muscle wrapping to high-resolution voxel-based finite element (FE) models. Such voxel-based models have a number of advantages over standard (geometry-based) FE models, but the increased resolution with which the load can be distributed over a model's surface is particularly advantageous, reflecting more closely how muscle fibre attachments are distributed. In this paper, the development, application and validation of a muscle wrapping method is illustrated using a simple cylinder. The algorithm: (1) calculates the shortest path over the surface of a bone given the points of origin and ultimate attachment of the muscle fibres; (2) fits a Non-Uniform Rational B-Spline (NURBS) curve from the shortest path and calculates its tangent, normal vectors and curvatures so that normal and tangential components of the muscle force can be calculated and applied along the fibre; and (3) automatically distributes the loads between adjacent fibres to cover the bone surface with a fully distributed muscle force, as is observed in vivo. Finally, we present a practical application of this approach to the wrapping of the temporalis muscle around the cranium of a macaque skull.

Functional relationship between skull form and feeding mechanics in Sphenodon, and implications for diapsid skull development.

The vertebrate skull evolved to protect the brain and sense organs, but with the appearance of jaws and associated forces there was a remarkable structural diversification. This suggests that the evolution of skull form may be linked to these forces, but an important area of debate is whether bone in the skull is minimised with respect to these forces, or whether skulls are mechanically "over-designed" and constrained by phylogeny and development. Mechanical analysis of diapsid reptile skulls could shed light on this longstanding debate. Compared to those of mammals, the skulls of many extant and extinct diapsids comprise an open framework of fenestrae (window-like openings) separated by bony struts (e.g., lizards, tuatara, dinosaurs and crocodiles), a cranial form thought to be strongly linked to feeding forces. We investigated this link by utilising the powerful engineering approach of multibody dynamics analysis to predict the physiological forces acting on the skull of the diapsid reptile Sphenodon. We then ran a series of structural finite element analyses to assess the correlation between bone strain and skull form. With comprehensive loading we found that the distribution of peak von Mises strains was particularly uniform throughout the skull, although specific regions were dominated by tensile strains while others were dominated by compressive strains. Our analyses suggest that the frame-like skulls of diapsid reptiles are probably optimally formed (mechanically ideal: sufficient strength with the minimal amount of bone) with respect to functional forces; they are efficient in terms of having minimal bone volume, minimal weight, and also minimal energy demands in maintenance.

Predicting muscle activation patterns from motion and anatomy: modelling the skull of Sphenodon (Diapsida: Rhynchocephalia).

The relationship between skull shape and the forces generated during feeding is currently under widespread scrutiny and increasingly involves the use of computer simulations such as finite element analysis. The computer models used to represent skulls are often based on computed tomography data and thus are structurally accurate; however, correctly representing muscular loading during food reduction remains a major problem. Here, we present a novel approach for predicting the forces and activation patterns of muscles and muscle groups based on their known anatomical orientation (line of action). The work was carried out for the lizard-like reptile Sphenodon (Rhynchocephalia) using a sophisticated computer-based model and multi-body dynamics analysis. The model suggests that specific muscle groups control specific motions, and that during certain times in the bite cycle some muscles are highly active whereas others are inactive. The predictions of muscle activity closely correspond to data previously recorded from live Sphenodon using electromyography. Apparent exceptions can be explained by variations in food resistance, food size, food position and lower jaw motions. This approach shows considerable promise in advancing detailed functional models of food acquisition and reduction, and for use in other musculoskeletal systems where no experimental determination of muscle activity is possible, such as in rare, endangered or extinct species.