Search

Hallux valgus (HV), the bunion of the first metatarsophalangeal joint (MTPJ), bothers many adults. No consensus has been reached about the causes of HV, be it a hereditary, or acquired, or multifactorial disease. Nor has agreement been reached using MTPJ angle to assess HV based on X-ray because in most cases the assessment of MTPJ is not reliable as it depends on the posture during scanning. In this study, we assume that HV is predominately acquired and that shoe wearing per se is an important player in HV pathogenesis. To verify our hypothesis, a CT-based finite element (FE) model of the first MTPJ of fossil remains of bear-footed Homo naledi was created and compared to that of five contemporary shoe-wearing wrestlers (10 models from two scans at an interval of about 18 months) because Homo naledi 's first MTPJ is an ideal model for non-shoe wearing with parallel sesamoid grooves. We developed the first MTPJ structure transformation method and created MTPJ joint capsule model for both Homo naledi and wrestlers. Constraint on the medial side of the first MTPJ capsule was set to simulate shoe-wearing conditions compared to the lack of medial constraint for barefooted conditions. Analysis of eight FE models of different angles for the first MTPJ of Homo naledi was performed by the first MTPJ transformation method and results showed that stress concentrated on the medial capsule of the first MTPJ in simulated shoe-wearing conditions, even at MTPJ angle of 0°. Increase in the first MTPJ angle further increased stress concentration on the medial side, and stress-growth relationship might reveal the causes of HV. We further developed a method to position the first MTPJ in wrestlers and created CT-based models at two time points. It was evident that the first MTPJ angle increased in all but one athlete, with a maximal increase of 4.03 degrees. This verifies our hypothesis that HV might be developed by wearing shoes. Further longitudinal studies with larger sample sizes are needed to additionally validate our results and determine the magnitude of the effects of shoe wearing on development and progression of HV., (Copyright © 2020 Yu, Fan, Fan, Li, Liu, Antonijevic, Milovanovic, Zhang, Li, Djuric and Fan.)

Electroencephalogram (EEG) analysis has been widely used in the diagnosis of stroke diseases for its low cost and noninvasive characteristics. In order to classify the EEG signals of stroke patients with cerebral infarction and cerebral hemorrhage, this paper proposes a novel EEG stroke signal classification method. This method has two highlights. The first is that a multi-feature fusion method is given by combining wavelet packet energy, fuzzy entropy and hierarchical theory. The second highlight is that a suitable classification model based on ensemble classifier is constructed for perfectly classification stroke signals. Entropy is an accessible way to measure information and uncertainty of time series. Many entropy-based methods have been developed these years. By comparing with the performances of permutation entropy, sample entropy, approximate entropy in measuring the characteristic of stroke patient's EEG signals, it can be found that fuzzy entropy has best performance in characterization stroke EEG signal. By combining hierarchical theory, wavelet packet energy and fuzzy entropy, a multi-feature fusion method is proposed. The method first calculates wavelet packet energy of EEG stroke signal, then extracts hierarchical fuzzy entropy feature by combining hierarchical theory and fuzzy entropy. The experimental results show that, compared with the fuzzy entropy feature, the classification accuracy based on the fusion feature of wavelet packet energy and hierarchical fuzzy entropy is much higher than benchmark methods. It means that the proposed multi-feature fusion method based on stroke EEG signal is an efficient measure in classifying ischemic and hemorrhagic stroke. Support vector machine (SVM), decision tree and random forest are further used as the stroke signal classification models for classifying ischemic stroke and hemorrhagic stroke. Experimental results show that, based on the proposed multi-feature fusion method, the ensemble method of random forest can get the best classification performance in accuracy among three models.

The detailed kinetics study of erythrocyte deformability is useful for the early diagnosis of blood diseases and for monitoring the blood rheology. Present solutions for a three-dimensional (3D) reconstruction of erythrocytes has a limited potential. This study aimed to use erythrocyte transmission electron images (ETIs) to evaluate the morphological relationship between adjacent ETIs and generate erythrocytes 3D model. First, ultrathin serial sections of skeletal muscle tissue were obtained using an ultramicrotome. Further, the set of ETIs in a capillary were captured by transmission electron microscopy. The images were aligned by translations and rotations using custom software to optimize the morphological relationship between adjacent ETIs. These coordinate transformations exploit the unique principal axis of inertia of each image to define the body coordinate system and hence provide the means to accurately reconnect the adjacent ETIs. The sum of the distances between the corresponding points on the boundary of adjacent ETIs was minimized and, further, was optimized by using physiological relationship between the adjacent ETIs. The analysis allowed to define precise virtual relationship between the adjacent erythrocytes. Finally, extracted erythrocytes' cross-section images allowed to generate 3D model of the erythrocytes.

Fed-batch culture is the most commonly used upstream process in industry today for recombinant monoclonal antibody production using Chinese hamster ovary (CHO) cells. Developing and optimizing this process in the lab is crucial for establishing process knowledge, which enables rapid and predictable tech-transfer to manufacturing scale. In this chapter, we describe stepwise how to carry out fed-batch CHO cell culture for lab-scale antibody production.

Cross-sectional imaging is considered the gold standard in diagnosing a range of diseases. However, despite its widespread use in clinical practice and research, no widely accepted method is available to reliably match cross-sectional planes in several consecutive scans. This deficiency can impede comparison between cross-sectional images and ultimately lead to misdiagnosis. Here, we propose and demonstrate a method for finding the same imaging plane in images obtained during separate scanning sessions. Our method is based on the reconstruction of a "virtual organ" from which arbitrary cross-sectional images can be extracted, independent of the axis orientation in the original scan or cut; the key is to establish unique body coordinates of the organ from its principal axes of inertia. To verify our method a series of tests were performed, and the same cross-sectional plane was successfully extracted. This new approach offers clinicians access, after just a single scanning session, to the morphology and structure of a lesion through cross-sectional images reconstructed along arbitrary axes. It also aids comparable detection of morphological and structural changes in the same imaging plane from scans of the same patient taken at different times-thus potentially reducing the misdiagnosis rate when cross-sectional images are interpreted.

Chinese hamster ovary (CHO) cells have become the primary expression system for the production of complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans. Control and optimization of protein N-glycosylation is crucial, as the structure of N-glycans can largely influence both biological and physicochemical properties of recombinant proteins. Protein N-glycosylation in CHO cell culture can be controlled and tuned by engineering medium, feed, culture process, as well as genetic elements of the cell. In this chapter, we will focus on how to carry out experiments for N-glycosylation modulation through medium and feed optimization. The workflow and typical methods involved in the experiment process will be presented.

Ensuring consistent glycosylation-associated quality of therapeutic monoclonal antibodies (mAbs) has become a priority in pharmaceutical bioprocessing given that the distribution and composition of the carbohydrates (glycans) bound to these molecules determines their therapeutic efficacy and immunogenicity. However, the interaction between bioprocess conditions, cellular metabolism and the intracellular process of glycosylation remains to be fully understood. To gain further insight into these interactions, we present a novel integrated modelling platform that links dynamic variations in mAb glycosylation with cellular secretory capacity. Two alternative mechanistic representations of how mAb specific productivity (qp ) influences glycosylation are compared. In the first, mAb glycosylation is modulated by the linear velocity with which secretory cargo traverses the Golgi apparatus. In the second, glycosylation is influenced by variations in Golgi volume. Within our modelling framework, both mechanisms accurately reproduce experimentally-observed dynamic changes in mAb glycosylation. In addition, an optimisation-based strategy has been developed to estimate the concentration of glycosylation enzymes required to minimise mAb glycoform variability. Our results suggest that the availability of glycosylation machinery relative to cellular secretory capacity may play a crucial role in mAb glycosylation. In the future, the modelling framework presented here may aid in selecting and engineering cell lines that ensure consistent mAb glycosylatio., (Copyright © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Therapeutic monoclonal antibodies (mAbs) are mainly produced by heterologous expression in Chinese hamster ovary (CHO) cells. The glycosylation profile of the mAbs has major impact on the efficacy and safety of the drug and is therefore an important parameter to control during production. In this study, the effect on IgG N-glycosylation from feeding CHO cells with eight glycosylation precursors during cultivation was investigated. The study was conducted in fed-batch mode in bioreactors with biological replicates to obtain highly controlled and comparable conditions. We assessed charge heterogeneity and glycosylation patterns of IgG. None of the eight feed additives caused statistically significant changes to cell growth or IgG productivity, compared to controls. However, the addition of 20 mM galactose did result in a reproducible increase of galactosylated IgG from 14% to 25%. On the other hand, addition of 20 mM N-acetyl-D-glucosamine (GlcNAc) reduced relative abundance of galactosylated IgG by 4%. Additionally, supplementation with 10 mM mannose slightly reduced GlcNAc occupancy of IgG. Overall, comparing the effects of IgG glycosylation, by supplementing the cell culture medium with glycosylation precursors during cultivation, revealed an application of these glycosylation precursors for modulating N-glycosylation of IgG., (© 2015 Wiley Periodicals, Inc.)

In this study, omics-based analysis tools were used to explore the effect of glucose starvation and culture duration on monoclonal antibody (mAb) production in fed-batch CHO cell culture to gain better insight into how these parameters can be controlled to ensure optimal mAb productivity and quality. Titer and N-glycosylation of mAbs, as well as proteomic signature and metabolic status of the production cells in the culture were assessed. We found that the impact of glucose starvation on the titer and N-glycosylation of mAbs was dependent on the degree of starvation during early stationary phase of the fed-batch culture. Higher degree of glucose starvation reduced intracellular concentrations of UDP-GlcNAc and UDP-GalNAc, but increased the levels of UDP-Glc and UDP-Gal. Increased GlcNAc and Gal occupancy correlated well with increased degree of glucose starvation, which can be attributed to the interplay between the dilution effect associated with change in specific productivity of mAbs and the changed nucleotide sugar metabolism. Herein, we also show and discuss that increased cell culture duration negatively affect the maturation of glycans. In addition, comparative proteomics analysis of cells was conducted to observe differences in protein abundance between early growth and early stationary phases. Generally higher expression of proteins involved in regulating cellular metabolism, extracellular matrix, apoptosis, protein secretion and glycosylation was found in early stationary phase. These analyses offered a systematic view of the intrinsic properties of these cells and allowed us to explore the root causes correlating culture duration with variations in the productivity and glycosylation quality of monoclonal antibodies produced with CHO cells., (© 2015 Wiley Periodicals, Inc.)

Fed-batch Chinese hamster ovary (CHO) cell culture is the most commonly used process for IgG production in the biopharmaceutical industry. Amino acid and glucose consumption, cell growth, metabolism, antibody titer, and N-glycosylation patterns are always the major concerns during upstream process optimization, especially media optimization. Gaining knowledge on their interrelations could provide insight for obtaining higher immunoglobulin G (IgG) titer and better controlling glycosylation-related product quality. In this work, different fed-batch processes with two chemically defined proprietary media and feeds were studied using two IgG-producing cell lines. Our results indicate that the balance of glucose and amino acid concentration in the culture is important for cell growth, IgG titer and N-glycosylation. Accordingly, the ideal fate of glucose and amino acids in the culture could be mainly towards energy and recombinant product, respectively. Accumulation of by-products such as NH4(+) and lactate as a consequence of unbalanced nutrient supply to cell activities inhibits cell growth. The levels of Leu and Arg in the culture, which relate to cell growth and IgG productivity, need to be well controlled. Amino acids with the highest consumption rates correlate with the most abundant amino acids present in the produced IgG, and thus require sufficient availability during culture. Case-by-case analysis is necessary for understanding the effect of media and process optimization on glycosylation. We found that in certain cases the presence of Man5 glycan can be linked to limitation of UDP-GlcNAc biosynthesis as a result of insufficient extracellular Gln. However, under different culture conditions, high Man5 levels can also result from low α-1,3-mannosyl-glycoprotein 2-β-N-acetylglucosaminyltransferase (GnTI) and UDP-GlcNAc transporter activities, which may be attributed to high level of NH4+ in the cell culture. Furthermore, galactosylation of the mAb Fc glycans was found to be limited by UDP-Gal biosynthesis, which was observed to be both cell line and cultivation condition-dependent. Extracellular glucose and glutamine concentrations and uptake rates were positively correlated with intracellular UDP-Gal availability. All these findings are important for optimization of fed-batch culture for improving IgG production and directing glycosylation quality., (© 2014 Wiley Periodicals, Inc.)