Global transcriptome profiles provide insights into muscle cell development and differentiation on microstructured marine biopolymer scaffolds for cultured meat production.

Biomaterial scaffolds play a pivotal role in the advancement of cultured meat technology, facilitating essential processes like cell attachment, growth, specialization, and alignment. Currently, there exists limited knowledge concerning the creation of consumable scaffolds tailored for cultured meat applications. This investigation aimed to produce edible scaffolds featuring both smooth and patterned surfaces, utilizing biomaterials such as salmon gelatin, alginate, agarose and glycerol, pertinent to cultured meat and adhering to food safety protocols. The primary objective of this research was to uncover variations in transcriptomes profiles between flat and microstructured edible scaffolds fabricated from marine-derived biopolymers, leveraging high-throughput sequencing techniques. Expression analysis revealed noteworthy disparities in transcriptome profiles when comparing the flat and microstructured scaffold configurations against a control condition. Employing gene functional enrichment analysis for the microstructured versus flat scaffold conditions yielded substantial enrichment ratios, highlighting pertinent gene modules linked to the development of skeletal muscle. Notable functional aspects included filament sliding, muscle contraction, and the organization of sarcomeres. By shedding light on these intricate processes, this study offers insights into the fundamental mechanisms underpinning the generation of muscle-specific cultured meat.

Systemic Inflammation but not Oxidative Stress Is Associated with Physical Performance in Moderate Chronic Obstructive Pulmonary Disease.

Chronic obstructive pulmonary disease (COPD) patients manifest muscle dysfunction and impaired muscle oxidative capacity, which result in reduced exercise capacity and poor health status. The aim of this study was to compare the physical performance, systemic inflammation, and oxidative stress of patients with moderate COPD, and to associate physical performance with inflammatory and oxidative stress plasma markers. Twenty CONTROL (n = 10) and moderate COPD (n = 10) patients participated in this study. Systematic inflammation and oxidative stress plasma markers, maximal aerobic capacity (VO2peak), and maximal isometric strength (MVIC) of the knee extensor (KE) muscles were measured. VO2peak was 31.3% greater in CONTROL compared to COPD (P = 0.006). The MVIC strength of the KE was 43.9% greater in CONTROL compared to COPD (P = 0.002). Tumor necrosis factor-alpha (TNF-α) was 79.6% greater in COPD compared to CONTROL (P < 0.001). Glutathione peroxidase activity (GPx) activity was 27.5% lesser in COPD compared to CONTROL (P = 0.05). TNF-α concentration was correlated with KE MVC strength (R = -0.48; P = 0.045) and VO2peak (R = -0.58; P = 0.01). Meanwhile, malondialdehyde (MDA) and GPx activity were not associated with KE strength or VO2peak (P = 0.74 and P = 0.14, respectively). COPD patients showed lesser muscle strength and aerobic capacity than healthy control individuals. Furthermore, patients with COPD showed greater systemic inflammation and lesser antioxidant capacity than healthy counterparts. A moderate association was evident between levels of systemic inflammation and physical performance variables.

Propagation of asymmetric optical vortex beams through turbulence and evolution of their OAM spectra.

In the realm of wave propagation through turbulent media, the spectrum of the orbital angular momentum of optical vortex beams is known to undergo symmetric broadening. However, the evolution of beams that are initially azimuthally asymmetric represents a distinct phenomenon. In this work, we have developed an analytical model describing the propagation of asymmetric OAM beams through the so-called Kolmogorov turbulence. Our results describe how the perturbation strength and the initial beam properties lead to a nonsymmetric spectrum of OAM modes. These findings lay the groundwork for further use of asymmetric fields that propagate in inhomogeneous media and their applications such as communications and sensing.

Micropatterned Nanofiber Scaffolds of Salmon Gelatin, Chitosan, and Poly(vinyl alcohol) for Muscle Tissue Engineering.

The development of scaffolds that mimic the aligned fibrous texture of the extracellular matrix has become an important requirement in muscle tissue engineering. Electrospinning is a widely used technique to fabricate biomimetic scaffolds. Therefore, a biopolymer blend composed of salmon gelatin (SG), chitosan (Ch), and poly(vinyl alcohol) (PVA) was developed by electrospinning onto a micropatterned (MP) collector, resulting in a biomimetic scaffold for seeding muscle cells. Rheology and surface tension studies were performed to determine the optimum solution concentration and viscosity for electrospinning. The scaffold microstructure was analyzed using SEM to determine the nanofiber's diameter and orientation. Blends of SG/Ch/PVA exhibited better electrospinnability and handling properties than pure PVA. The resulting scaffolds consist of a porous surface (∼46%), composed of a random fiber distribution, for a flat collector and scaffolds with regions of aligned nanofibers for the MP collector. The nanofiber diameters are 141 ± 2 and 151 ± 2 nm for the flat and MP collector, respectively. In vitro studies showed that myoblasts cultured on scaffold SG/Ch/PVA presented a high rate of cell growth. Furthermore, the aligned nanofibers on the SG/Ch/PVA scaffold provide a suitable platform for myoblast alignment.

Effect of Electrical Stimulation on PC12 Cells Cultured in Different Hydrogels: Basis for the Development of Biomaterials in Peripheral Nerve Tissue Engineering.

Extensive damage to peripheral nerves is a health problem with few therapeutic alternatives. In this context, the development of tissue engineering seeks to obtain materials that can help recreate environments conducive to cellular development and functional repair of peripheral nerves. Different hydrogels have been studied and presented as alternatives for future treatments to emulate the morphological characteristics of nerves. Along with this, other research proposes the need to incorporate electrical stimuli into treatments as agents that promote cell growth and differentiation; however, no precedent correlates the simultaneous effects of the types of hydrogel and electrical stimuli. This research evaluates the neural differentiation of PC12 cells, relating the effect of collagen, alginate, GelMA, and PEGDA hydrogels with electrical stimulation modulated in four different ways. Our results show significant correlations for different cultivation conditions. Electrical stimuli significantly increase neural differentiation for specific experimental conditions dependent on electrical frequency, not voltage. These backgrounds allow new material treatment schemes to be formulated through electrical stimulation in peripheral nerve tissue engineering.

Use of Chitosan from Southern King Crab to Develop Films Functionalized with RGD Peptides for Potential Tissue Engineering Applications.

Southern King Crab (SKC) represents an important fishery resource that has the potential to be a natural source of chitosan (CS) production. In tissue engineering, CS is very useful to generate biomaterials. However, CS has a lack of signaling molecules that facilitate cell-substrate interaction. Therefore, RGD (arginine-glycine-aspartic acid) peptides corresponding to the main integrin recognition site in extracellular matrix proteins have been used to improve the CS surface. The aim of this study was to evaluate in vitro cell adhesion and proliferation of CS films synthesized from SKC shell wastes functionalized with RGD peptides. The FTIR spectrum of CS isolated from SKC shells (SKC-CS) was comparable to commercial CS. Thermal properties of films showed similar endothermic peaks at 53.4 and 53.0 °C in commercial CS and SKC-CS, respectively. The purification and molecular masses of the synthesized RGD peptides were confirmed using HPLC and ESI-MS mass spectrometry, respectively. Mouse embryonic fibroblast cells showed higher adhesion on SKC-CS (1% w/v) film when it was functionalized with linear RGD peptides. In contrast, a cyclic RGD peptide showed similar adhesion to control peptide (RDG), but the highest cell proliferation was after 48 h of culture. This study shows that functionalization of SKC-CS films with linear or cyclic RGD peptides are useful to improve effects on cell adhesion or cell proliferation. Furthermore, our work contributes to knowledge of a new source of CS to synthesize constructs for tissue engineering applications.

First-order statistics of intensity and phase in Laguerre-Gauss speckles.

Laguerre-Gaussian (LG) beams are characterized by an azimuthal index or topological charge (m), associated with the orbital angular momentum, and by a radial index (p), which represents the number of the rings in the intensity distribution. We present a detailed, systematic study of the first-order phase statistics of the speckle fields created when LG beams of different order interact with random phase screens with different optical roughness. The phase properties of the LG speckle fields are studied in both the Fresnel and the Fraunhofer regimes using the equiprobability density ellipse formalism such that analytical expressions can be derived for the phase statistics.

Enhanced recombinant protein production in CHO cell continuous cultures under growth-inhibiting conditions is associated with an arrested cell cycle in G1/G0 phase.

Low temperature and sodium butyrate (NaBu) are two of the most used productivity-enhancing strategies in CHO cell cultures during biopharmaceutical manufacturing. While these two approaches alter the balance in the reciprocal relationship between cell growth and productivity, we do not fully understand their mechanisms of action beyond a gross cell growth inhibition. Here, we used continuous culture to evaluate the differential effect of low temperature and NaBu supplementation on CHO cell performance and gene expression profile. We found that an increase in cell-productivity under growth-inhibiting conditions was associated with the arrest of cells in the G1/G0 phase. A transcriptome analysis revealed that the molecular mechanisms by which low temperature and NaBu arrested cell cycle in G1/G0 differed from each other through the deregulation of different cell cycle checkpoints and regulators. The individual transcriptome changes in pattern observed in response to low temperature and NaBu were retained when these two strategies were combined, leading to an additive effect in arresting the cell cycle in G1/G0 phase. The findings presented here offer novel molecular insights about the cell cycle regulation during the CHO cell bioprocessing and its implications for increased recombinant protein production. This data provides a background for engineering productivity-enhanced CHO cell lines for continuous manufacturing.

Phase memory of optical vortex beams.

Optical vortex beams are under considerable scrutiny due to their demonstrated potential for applications ranging from quantum optics to optical communications and from material processing to particle trapping. However, upon interaction with inhomogeneous material systems, their deterministic properties are altered. The way these structured beams are affected by different levels of disturbances is critical for their uses. Here, for the first time, we quantify the degradation of perfect optical vortex beams after their interaction with localized random media. We developed an analytical model that (1) describes how the spatial correlation and the phase variance of disturbance affect the phase distribution across the vortex beams and (2) establishes the regimes of randomness for which the beams maintain the memory of their initial vorticity. Systematic numerical simulations and controlled experiments demonstrate the extent of this memory effect for beams with different vorticity indices.

Osteoconductive Effect of a Nanocomposite Membrane Treated with UV Radiation.

Modulation of the bio-regenerative characteristics of materials is an indispensable requirement in tissue engineering. Particularly, in bone tissue engineering, the promotion of the osteoconductive phenomenon determines the elemental property of a material be used therapeutically. In addition to the chemical qualities of the constituent materials, the three-dimensional surface structure plays a fundamental role that various methods are expected to modulate in a number of ways, one most promising of which is the use of different types of radiation. In the present manuscript, we demonstrate in a calvarial defect model, that treatment with ultraviolet irradiation allows modification of the osteoconductive characteristics in a biomaterial formed by gelatin and chitosan, together with the inclusion of hydroxyapatite and titanium oxide nanoparticles.

3D intensity correlations in random fields created by vortex structured beams.

We develop an analytical model for the 3D spatial coherence function of speckle fields generated by scattering of vortex and perfect optical vortex beams. The model is general and describes the spatial coherence along both the transversal and the longitudinal directions. We found that, on propagation, the 3D spatial coherence evolves differently for the different types of initially structured beams, which may affect their use in a variety of sensing applications.

Colloidal density control with Bessel-Gauss beams.

Optical manipulation of colloidal systems is of high interest for both fundamental studies and practical applications. It has been shown that optically induced thermophoresis and nonlinear interactions can significantly affect the properties of dense colloidal media. However, macroscopic scale phenomena can also be generated at thermal equilibrium. Here, we demonstrate that steady-state variations of particle density can be created over large, three-dimensional regions by appropriately structured external optical fields. We prove analytically and experimentally that an optical vortex beam can dynamically control the spatial density of microscopic particles along the direction of its propagation. We show that these artificial steady-states can be generated at will and can be maintained indefinitely, which can be beneficial for applications such as path clearing and mass transportation.

A New Insight for the Identification of Oncogenic Variants in Breast and Prostate Cancers in Diverse Human Populations, With a Focus on Latinos.

Background: Breast cancer (BRCA) and prostate cancer (PRCA) are the most commonly diagnosed cancer types in Latin American women and men, respectively. Although in recent years large-scale efforts from international consortia have focused on improving precision oncology, a better understanding of genomic features of BRCA and PRCA in developing regions and racial/ethnic minority populations is still required. Methods: To fill in this gap, we performed integrated in silico analyses to elucidate oncogenic variants from BRCA and PRCA driver genes; to calculate their deleteriousness scores and allele frequencies from seven human populations worldwide, including Latinos; and to propose the most effective therapeutic strategies based on precision oncology. Results: We analyzed 339,100 variants belonging to 99 BRCA and 82 PRCA driver genes and identified 18,512 and 15,648 known/predicted oncogenic variants, respectively. Regarding known oncogenic variants, we prioritized the most frequent and deleterious variants of BRCA (n = 230) and PRCA (n = 167) from Latino, African, Ashkenazi Jewish, East Asian, South Asian, European Finnish, and European non-Finnish populations, to incorporate them into pharmacogenomics testing. Lastly, we identified which oncogenic variants may shape the response to anti-cancer therapies, detailing the current status of pharmacogenomics guidelines and clinical trials involved in BRCA and PRCA cancer driver proteins. Conclusion: It is imperative to unify efforts where developing countries might invest in obtaining databases of genomic profiles of their populations, and developed countries might incorporate racial/ethnic minority populations in future clinical trials and cancer researches with the overall objective of fomenting pharmacogenomics in clinical practice and public health policies.

Anatase Incorporation to Bioactive Scaffolds Based on Salmon Gelatin and Its Effects on Muscle Cell Growth.

The development of new polymer scaffolds is essential for tissue engineering and for culturing cells. The use of non-mammalian bioactive components to formulate these materials is an emerging field. In our previous work, a scaffold based on salmon gelatin was developed and tested in animal models to regenerate tissues effectively and safely. Here, the incorporation of anatase nanoparticles into this scaffold was formulated, studying the new composite structure by scanning electron microscopy, differential scanning calorimetry and dynamic mechanical analysis. The incorporation of anatase nanoparticles modified the scaffold microstructure by increasing the pore size from 208 to 239 µm and significantly changing the pore shape. The glass transition temperature changed from 46.9 to 55.8 °C, and an increase in the elastic modulus from 79.5 to 537.8 kPa was observed. The biocompatibility of the scaffolds was tested using C2C12 myoblasts, modulating their attachment and growth. The anatase nanoparticles modified the stiffness of the material, making it possible to increase the growth of myoblasts cultured onto scaffolds, which envisions their use in muscle tissue engineering.

First-order statistics of the phase in optical vortex speckles.

We present the theoretical analysis of first-order statistics of the phase in a far-field speckle field, which originates from an optical vortex passing through a random phase screen. By using the concept of the equiprobability density ellipse, we show that the standard deviation of the phase in a speckle field varies non-monotonically in the radial direction and, more interestingly, it exhibits a minimum at a certain radial position determined by the topological charge. In the limit of zero topological charge, the phase statistics naturally converges to the expectation corresponding to the incident Gaussian beam.

A New Edible Film to Produce In Vitro Meat.

In vitro meat is a novel concept of food science and biotechnology. Methods to produce in vitro meat employ muscle cells cultivated on a scaffold in a serum-free medium using a bioreactor. The microstructure of the scaffold is a key factor, because muscle cells must be oriented to generate parallel alignments of fibers. This work aimed to develop a new scaffold (microstructured film) to grow muscle fibers. The microstructured edible films were made using micromolding technology. A micromold was tailor-made using a laser cutting machine to obtain parallel fibers with a diameter in the range of 70-90 µm. Edible films were made by means of solvent casting using non-mammalian biopolymers. Myoblasts were cultured on flat and microstructured films at three cell densities. Cells on the microstructured films grew with a muscle fiber morphology, but in the case of using the flat film, they only produced unorganized cell proliferation. Myogenic markers were assessed using quantitative polymerase chain reaction. After 14 days, the expression of desmin, myogenin, and myosin heavy chain were significantly higher in microstructured films compared to the flat films. The formation of fiber morphology and the high expression of myogenic markers indicated that a microstructured edible film can be used for the production of in vitro meat.

Cold-adaptation of a methacrylamide gelatin towards the expansion of the biomaterial toolbox for specialized functionalities in tissue engineering.

Tissue regeneration is witnessing a significant surge in advanced medicine. It requires the interaction of scaffolds with different cell types for efficient tissue formation post-implantation. The presence of tissue subtypes in more complex organs demands the co-existence of different biomaterials showing different hydrolysis rate for specialized cell-dependent remodeling. To expand the available toolbox of biomaterials with sufficient mechanical strength and variable rate of enzymatic degradation, a cold-adapted methacrylamide gelatin was developed from salmon skin. Compared with mammalian methacrylamide gelatin (GelMA), hydrogels derived from salmon GelMA displayed similar mechanical properties than the former. Nevertheless, salmon gelatin and salmon GelMA-derived hydrogels presented characteristics common of cold-adaptation, such as reduced activation energy for collagenase, increased enzymatic hydrolysis turnover of hydrogels, increased interconnected polypeptides molecular mobility and lower physical gelation capability. These properties resulted in increased cell-remodeling rate in vitro and in vivo, proving the potential and biological tolerance of this mechanically adequate cold-adapted biomaterial as alternative scaffold subtypes with improved cell invasion and tissue fusion capacity.

Spatial intensity correlations of a vortex beam and a perfect optical vortex beam.

We present a model based on the Fresnel diffraction scheme for the spatial coherence function of random fields created by scattering optical vortex and perfect vortex beams. By using the spatial coherence function we showed analytically, numerically, and experimentally the dependence and independence of the speckle size of an optical vortex and perfect optical vortex (POV) with a topological charge, respectively. We also showed in both cases the linear dependence of speckle size on the distance of propagation. Furthermore, we describe a regime in which the spatial coherence function is nonevolving for the optical vortex beam and the POV beam with the propagation distance.

The role of phase I and II genetic polymorphisms, smoking, alcohol and cancer family history, in the risk of developing testicular cancer.

BACKGROUND: Testicular cancer (TCa) is a malignant tumor with highest incidence and mortality rates in Chile. The genes coding for cytochrome P450, glutathione-S-transferases (GSTs), and UDP glucuronyl transferases (UGT) participate in the biotransformation and detoxification of xenobiotics. Mutations in these genes have been associated with a high incidence of various types of cancer and an increased risk of presenting adverse reactions to drugs. OBJECTIVE: The aim of this study was to relate the presence of genetic polymorphisms in cytochrome P450 1A1 (CYP1A1), CYP3A4, GSTM1, GSTP1, GSTT1, and UGT1A1 genes and nongenetic factors with the risk of developing TCa. METHODS: A total of 276 volunteers from the Chilean general population and 251 Chilean TCa patients were recruited for the study. Genotypic analyses were performed using qPCR and PCR-RFLP. RESULTS: Variant alleles found to increase the risk of developing TCa were CYP1A1*2C (rs1048943), GSTP1 (rs1695), and GSTT1null, especially when in conjunction with a cancer family history and/or a smoking habit. The results of the multivariate analysis showed that the presence of variant alleles of GSTP1 (rs1695) together with a smoking habit and a family history of cancer accounted for a 15.9% risk of developing TCa in the Chilean population. CYP1A1*2C, GSTM1null, GSTT1null, and GSTP1 (rs1695) are statistically related to the risk of appearance of TCa, alone or associated with nongenetic factors. CONCLUSION: Therefore, phase I and II variant alleles might be useful in evaluating susceptibility to TCa in the studied population.

Glass Transition in Crosslinked Nanocomposite Scaffolds of Gelatin/Chitosan/Hydroxyapatite.

The development of biopolymeric scaffolds crosslinked with nanoparticles is an emerging field. Gelatin/chitosan scaffolds are gaining interest in medical areas, e.g., bone tissue engineering, given their suitability for nano-hydroxyapatite incorporation. The glass transition temperature is a thermodynamic property of polymer scaffolds that changes with crosslinker or nanofiller concentration. Here, we report the experimental change in glass transition temperature of gelatin/chitosan scaffolds modified by hydroxyapatite nanoparticles and crosslinker concentration. Our results show synergic effects between nanoparticles and crosslinking, which leads to a non-linear behavior of the glass transition temperature. Furthermore, a theoretical model to predict glass transition is proposed. This model can be used as a mathematical tool for the design of future scaffolds used in bone tissue engineering.