3D observations provide striking findings in rubber elasticity.

The mechanical response of rubbers has been ubiquitously assumed to be only a function of the imposed strain. Using innovative X-ray measurements capturing the three-dimensional spatial volumetric strain fields, we demonstrate that rubbers and indeed many common engineering polymers undergo significant local volume changes. But remarkably, the overall specimen volume remains constant regardless of the imposed loading. This strange behavior which also leads to apparent negative local bulk moduli is due to the presence of a mobile phase within these materials. Combining X-ray tomographic observations with high-speed radiography to track the motion of the mobile phase, we have revised classical thermodynamic frameworks of rubber elasticity. The work opens broad avenues to understand not only the mechanical behavior of rubbers but a large class of widely used engineering polymers.

Exploiting the Biophysical Cues toward Dual Differentiation of hMSC's within Geometrical Patterns.

A wide variety of cues from the extracellular matrix (ECM) have been known to affect the differentiation of stem cells in vivo. In particular, the biophysical cues and cell shape have been known to affect the stem cell function, yet very little is known about the interplay between how these cues control differentiation. For the first time, by using photolithography to pattern poly(ethylene glycol) (PEG), patterns of square and triangular geometries were created, and the effect of these structures and the biophysical cues arising were utilized to differentiate cells into multiple lineages inside a same pattern without the use of any adhered protein or growth factors. The data from these studies showed that the cells present at the edges were well elongated, exhibit high aspect ratios, and differentiated into osteogenic lineage, whereas the cells present at the center exhibit lower aspect ratio and were primarily adipogenic lineage regardless of the geometry. This was correlated to the higher expression of focal adhesion proteins at the edges, the expression of which have been known to affect the osteogenic differentiation. By showing MSC lineage commitment relationships due to physical signals, this study highlights the importance of these cues and cell shape in further understanding stem cell behavior for tissue engineering applications.

Single-shot multispectral quantitative phase imaging of biological samples using deep learning.

Multispectral quantitative phase imaging (MS-QPI) is a high-contrast label-free technique for morphological imaging of the specimens. The aim of the present study is to extract spectral dependent quantitative information in single-shot using a highly spatially sensitive digital holographic microscope assisted by a deep neural network. There are three different wavelengths used in our method: λ=532, 633, and 808 nm. The first step is to get the interferometric data for each wavelength. The acquired datasets are used to train a generative adversarial network to generate multispectral (MS) quantitative phase maps from a single input interferogram. The network was trained and validated on two different samples: the optical waveguide and MG63 osteosarcoma cells. Validation of the present approach is performed by comparing the predicted MS phase maps with numerically reconstructed (F T+T I E) phase maps and quantifying with different image quality assessment metrices.

Actinomyces ruminis sp. nov., an obligately anaerobic bacterium isolated from the rumen of cattle.

An obligately anaerobic, rod-shaped, Gram-stain-positive, non-spore-forming, non-motile bacterial strain; designated as CtC72T was isolated from the rumen of cattle. The 16S rRNA gene sequence similarity of less than 98.65% revealed the strain as a member of the genus Actinomyces, nearest to but distinct from Actinomyces qiguomingii DSM 106201T, Actinomyces ruminicola DSM 27982T, Actinomyces procaprae JCM 33484T, Actinomyces succiniciruminis TISTR 2317, Actinomyces glycerinitolerans TISTR 2318. The low values of digital DNA-DNA hybridization (< 70%) and average nucleotide identity (< 95%) further highlighted the distinctive nature of strain CtC72T from its closest relatives. The strain CtC72T could grow at temperatures between 30 and 50 °C (optimum 40 °C), pH between 6.0 and 9.0 (optimum 7.5-8.0), and NaCl between 0 and 1.5% (optimum 0%). The strain hydrolysed cellulose and xylan and utilised a range of mono-, di-, and oligo-saccharides as a source of carbon and energy. Glucose fermentation resulted in acetic acid and formic acid as major metabolic products, while propionic acid, lactic acid, and ethanol as minor products along with CO2 production. The DNA G + C content of strain CtC72T was 68.40 (mol%, Tm) and 68.05 (%, digital). Major cellular fatty acids (> 10%) were C16:0, C18:1 ω9c, and C18:1 ω9c DMA. Based on these data, we propose that strain CtC72T be classified as a novel species, Actinomyces ruminis sp. nov., under the genus Actinomyces. The type strain is CtC72T (= KCTC 15726T = JCM 32641T = MCC 3500T).

Exploiting Substrate Cues for Co-Culturing Cells in a Micropattern.

Spatial distribution of cells and their interactions between neighboring cells in native microenvironments are of fundamental importance in determining cell fate decisions such as migration, growth, and differentiation. Controlling the spatial distribution of different cell types in defined geometries can replicate these native environments, which can be a useful model for several studies. While spatiotemporal control over multiple cell arrangements is required to achieve the complex tissue architecture, unfortunately, conventional cell patterning techniques usually allow only single patterning with a single cell type. In the present study, we introduce a simple lithographic method to pattern multiple cell types in a spatially controlled manner by utilizing the biophysical cues present at the corners of the patterned geometry. By fabricating micropatterns of different shapes, we demonstrate how the cell can be constrained to pattern along the corners of patterned geometries owing to the presence of topographical cues, leaving empty voids in the center that can be further utilized for patterning a second cell type. We also demonstrate that the cell alignment along the pattern is a dynamic process and the cells migrate from a more uniform cell-adhesive region toward the topographical cues. The cytoskeleton arrangement was geometry-dependent, which was confirmed through a series of in vitro evaluations, such as scanning electron microscopy and fluorescence microscopy. These findings have not only helped us in exploring the importance of these cues in guiding the cell fate but have also allowed us to develop a technique, which self-patterns the cells without any expensive exogenous cues and can be used as a model protocol to eventually organize cells into a specific pattern with micron-scale precision in vitro.

Cultivation of multiple genera of hydrogenotrophic methanogens from different environmental niches.

Six genera of hydrogenotrophic methanogens, namely Methanobrevibacter, Methanobacterium, Methanocorpusculum, Methanothermobacter, Methanoculleus, and Methanospirillum were cultivated from diverse environmental niches like rumen, feces, gut, and sediments using BY medium. We also report a putative novel genus and two novel species of methanogens isolated from termite, Indian star tortoise, and green iguana.

Genetically engineering milk.

It has been thirty years since the first genetically engineered animal with altered milk composition was reported. During the intervening years, the world population has increased from 5bn to 7bn people. An increasing demand for protein in the human diet has followed this population expansion, putting huge stress on the food supply chain. Many solutions to the grand challenge of food security for all have been proposed and are currently under investigation and study. Amongst these, genetics still has an important role to play, aiming to continually enable the selection of livestock with enhanced traits. Part of the geneticist's tool box is the technology of genetic engineering. In this Invited Review, we indicate that this technology has come a long way, we focus on the genetic engineering of dairy animals and we argue that the new strategies for precision breeding demand proper evaluation as to how they could contribute to the essential increases in agricultural productivity our society must achieve.

Desymmetric homologating annulation to access chiral pentafulvenes and their application in bioimaging.

The architectural design of polycyclic/multisubstituted pentafulvenes has demonstrated great potential for the development of electrochromic materials and biologically active motifs. Unfortunately, the enantioselective construction of such distinctive cores with all carbon quaternary chiral centers has remained untouched to date. Herein, we disclose an enantioselective homologating annulation of cyclopent-4-ene-dione with 3-cyano-4-methylcoumarins through L-tert-leucine derived thiourea catalysis, affording a wide range of enantioenriched polycyclic multisubstituted embedded aminopentafulvenes with excellent stereocontrol (up to 99:1 er) and chemical yields up to 87%. A detailed photophysical and cytotoxicity analysis of racemic and chiral homologated adducts unveils the exceptional behavior of chiral adducts over their racemic analogs, highlighting the importance of stereoselectivity of the developed scaffolds. A cellular uptake experiment in a mammalian fibroblast cell line confirmed the potential of developed polycyclic aminopentafulvene cores as a highly promising labeling dye that can be utilized for bioimaging without any adverse effects.

Sensory deficits of the paretic and non-paretic upper limbs relate with the motor recovery of the poststroke subjects.

BACKGROUND: Post stroke, motor paresis has usually been considered to be a crucial factor responsible for the disability; other impairments such as somatosensory deficits may also play a role. OBJECTIVE: To determine the relation between the sensory deficits (paretic and non-paretic upper limbs) and the motor recovery of the paretic upper limb and to predict the potential of motor recovery based on the sensory deficits among stroke subjects. METHODS: The study was a cross-sectional study conducted in a rehabilitation institute. Ninety-five poststroke hemiparetic subjects having sensory impairment in any of the modalities were considered for this study. Sensory deficits were assessed on both the upper limbs (paretic and non-paretic) primarily using Erasmus MC modification of the revised version of Nottingham Sensory Assessment (Em-NSA) and Nottingham Sensory Assessment (Stereognosis) (NSA-S). The motor recovery was assessed using the Fugl-Meyer assessment (FMA). RESULTS: The measures of sensory deficits exhibited weak but significant correlation [the paretic (Em-NSA and NSA; r = .38 to .58; p < .001) and the non-paretic (Em-NSA and NSA; r = .24 to .38; p = .03 to .001)] with the motor recovery of the paretic upper limb as measured by FMA. The potential of favorable recovery of the paretic upper limb may be predicted using the cutoff scores of Em-NSA (30, 21, and 24) and NSA-S (5, 8, and 5) of the paretic side. CONCLUSION: In stroke, sensory deficits relate weakly with the recovery of the paretic upper limb and can predict recovery potential of the paretic upper limb.

Percutaneous Intervention of LVAD Outflow Graft Obstruction and Thrombosis.

Left ventricular assist devices serve as a salvage therapy for patients with advanced heart failure. Complications such as thrombosis and obstruction can lead to acute device malfunction, posing significant clinical risks. A multidisciplinary approach is crucial for management. Few cases in the literature have demonstrated the safety and efficacy of percutaneous intervention, which holds significant value due to its less invasive nature and minimal risk of morbidity, especially in high-risk surgical patients.

Anterior STEMI in a 25-year-old with Cogan syndrome.

Cogan syndrome (CS) is a chronic inflammatory disorder primarily affecting young adults. It is characterized by interstitial keratitis, vestibuloauditory dysfunction, and, rarely, systemic vasculitis and aortitis. In this case report, we present a rare instance of a young male with anterior ST-elevation myocardial infarction in the context of aneurysmal coronary vessels and proximal occlusion of the left anterior descending artery. This case underscores the importance of recognizing cardiac risk factors in the setting of CS, as they can be life-threatening. Regular follow up is necessary, especially for patients showing signs of vasculitis or experiencing cardiac symptoms. A multidisciplinary approach to management can help optimize the prognosis for these complex patients. Learning objective: Comprehend Cogan syndrome (CS): Grasp the clinical manifestations and potential cardiovascular complications of CS. Diagnostic approach in CS: Differentiate cardiac symptoms in CS from typical coronary diseases, emphasizing the role of heart catheterization. Multidisciplinary management: Recognize the importance of integrating rheumatologists, cardiologists, and surgeons for optimal CS patient outcomes.

Comprehensive Genetic Profiling Reveals Frequent Alterations of Driver Genes on the X Chromosome in Extranodal NK/T-cell Lymphoma.

Extranodal NK/T-cell lymphoma (ENKTCL) is an Epstein-Barr virus (EBV)-related neoplasm with male dominance and a poor prognosis. A better understanding of the genetic alterations and their functional roles in ENKTCL could help improve patient stratification and treatments. In this study, we performed a comprehensive genetic analysis of 178 ENKTCL cases to delineate the landscape of mutations, copy number alterations (CNA), and structural variations, identifying 34 driver genes including six previously unappreciated ones, namely, HLA-B, HLA-C, ROBO1, CD58, POT1, and MAP2K1. Among them, CD274 (24%) was the most frequently altered, followed by TP53 (20%), CDKN2A (19%), ARID1A (15%), HLA-A (15%), BCOR (14%), and MSN (14%). Chromosome X losses were the most common arm-level CNAs in females (∼40%), and alterations of four X-linked driver genes (MSN, BCOR, DDX3X, and KDM6A) were more frequent in males and females harboring chromosome X losses. Among X-linked drivers, MSN was the most recurrently altered, and its expression was lost in approximately one-third of cases using immunohistochemical analysis. Functional studies of human cell lines showed that MSN disruption promoted cell proliferation and NF-κB activation. Moreover, MSN inactivation increased sensitivity to NF-κB inhibition in vitro and in vivo. In addition, recurrent deletions were observed at the origin of replication in the EBV genome (6%). Finally, by integrating the 34 drivers and 19 significant arm-level CNAs, nonnegative matrix factorization and consensus clustering identified two molecular groups with different genetic features and prognoses irrespective of clinical prognostic factors. Together, these findings could help improve diagnostic and therapeutic strategies in ENKTCL. Significance: Integrative genetic analyses and functional studies in extranodal NK/T-cell lymphoma identify frequent disruptions of X-linked drivers, reveal prognostic molecular subgroups, and uncover recurrent MSN alterations that confer sensitivity to NF-κB inhibition.

Generation of Patterned Cocultures in 2D and 3D: State of the Art.

Cells inside the body are embedded into a highly structured microenvironment that consists of cells that lie in direct or close contact with other cell types that regulate the overall tissue function. Therefore, coculture models are versatile tools that can generate tissue engineering constructs with improved mimicking of in vivo conditions. While there are many reviews that have focused on pattering a single cell type, very few reviews have been focused on techniques for coculturing multiple cell types on a single substrate with precise control. In this regard, this Review covers various technologies that have been utilized for the development of these patterned coculture models while mentioning the limitations associated with each of them. Further, the application of these models to various tissue engineering applications has been discussed.

Three-dimensional full-field velocity measurements in shock compression experiments using stereo digital image correlation.

Shock compression plate impact experiments conventionally rely on point-wise velocimetry measurements based on laser-based interferometric techniques. This study presents an experimental methodology to measure the free surface full-field particle velocity in shock compression experiments using high-speed imaging and three-dimensional (3D) digital image correlation (DIC). The experimental setup has a temporal resolution of 100 ns with a spatial resolution varying from 90 to 200 µm/pixel. Experiments were conducted under three different plate impact configurations to measure spatially resolved free surface velocity and validate the experimental technique. First, a normal impact experiment was conducted on polycarbonate to measure the macroscopic full-field normal free surface velocity. Second, an isentropic compression experiment on Y-cut quartz-tungsten carbide assembly is performed to measure the particle velocity for experiments involving ramp compression waves. To explore the capability of the technique in multiaxial loading conditions, a pressure shear plate impact experiment was conducted to measure both the normal and transverse free surface velocities under combined normal and shear loading. The velocities measured in the experiments using digital image correlation are validated against previous data obtained from laser interferometry. Numerical simulations were also performed using established material models to compare and validate the experimental velocity profiles for these different impact configurations. The novel ability of the employed experimental setup to measure full-field free surface velocities with high spatial resolutions in shock compression experiments is demonstrated for the first time in this work.

Light-Mediated 3D Printing of Micro-Pyramid-Decorated Tailorable Wound Dressings with Endogenous Growth Factor Sequestration for Improved Wound Healing.

Medical dressings play an important role in the field of tissue engineering owing to their ability to accelerate the process of wound healing. Great efforts have been made to fabricate wound dressings with distinctive features for promoting wound healing. However, most of the current synthesis methods either generate dressings of uniform size or involve complex fabrication techniques, thus limiting their commercialization for the personalized dressings. We report here a dressing, which presents a paradigm shift in the design of the dressing from uniform films to a micro-patterned film. The hypothesis driving the design is the ability of the 3D patterns to provide an efficient transient matrix filling the depth of the wound rather than just providing a barrier and slight re-epithelialization. We demonstrate the use of the digital light processing 3D printing technique to generate micro-pyramid-decorated wound healing dressings with individualized design and with bio-compatible gelatin methacryloyl to contact the wounded areas. In addition to providing better adhesion to the migratory cells, the micro-pyramids also enable covalent conjugation of heparin, providing capability to sequester endogenous growth factors (GFs). Based on these advantages, the developed dressing not only adheres strongly to the wound bed but also promotes the treatment of a rat wound model by utilizing the power of endogenous GFs for tissue regeneration. Thus, it is believed that the developed dressing can break through the limitation of traditional wound treatment and be an ideal candidate for wound healing.

Healing of Endosseous Implants Having Different Surface Characteristics in the Alveolar Bone: A Clinical Study.

Background Total treatment time in implant placement can be significantly reduced by placing immediate implants into the freshly extracted sockets. Also, immediate implant placement can act as a guide for proper and accurate implant placement. Additionally, in immediate implant placement, the resorption of bone associated with the healing of the extraction socket is also reduced. This clinical study aimed to clinically and radiographically assess the healing of endosseous implants having different surface characteristics in nongrafted and grafted bone. Methodology In 68 subjects, 198 implants were placed, including 102 oxidized (TiUnite, Göteborg, Sweden) and 96 turned surface implants (Nobel Biocare Mark III, Göteborg) were placed. Survival was considered with clinical stability and acceptable function with no discomfort and no radiographic or clinical signs of pathology/infection. Rest cases that showed no healing and implant no osseointegration were considered failures. Clinical and radiographic examination was done by two experts after two years of loading based on bleeding on probing (BOP) mesially and distally, radiographic marginal bone levels, and probing depth (mesial and distal). Results Five implants failed in total where four implants were with the turned surface (Nobel Biocare Mark III) and one was from the oxidized surface (TiUnite). The one oxidized implant was in a 62-year-old female and was placed in the region of mandibular premolar (44) of length 13 mm and was lost within five months of placement before functional loading. Mean probing depth had a nonsignificant difference between oxidized and turned surfaces with the mean values of 1.6 ± 1.2 and 1.5 ± 1.0 mm, respectively, with P = 0.5984; mean BOP in oxidized and turned surfaces was 0.3 ± 0.7 and 0.4 ± 0.6, respectively (P = 0.3727). Marginal bone levels, respectively, were 2.0 ± 0.8 and 1.8 ± 0.7 mm (P = 0.1231). In marginal bone levels related to implant loading, a nonsignificant difference was seen in early loading and one-stage loading with P-values of 0.06 and 0.09, respectively. However, in two-stage placement, significantly higher values were seen for oxidized surfaces (2.4 ± 0.8 mm) compared to turned surfaces (1.9 ± 0.8 mm), with P = 0.0004. Conclusions This study concludes that nonsignificantly higher survival rates are associated with oxidized surfaces compared to turned surfaces after two years of follow-up. Higher marginal bone levels were seen in oxidized surfaces for single implants and implants placed in two stages.

Natural cocktail of bioactive factors conjugated on nanofibrous dressing for improved wound healing.

Any interference in the timely and orderly progression through all the phases of healing process can turn a minor injury into a chronic wound. Most of the wound dressings available in the market are moderately effective and have not shown satisfactory improvement in healing. Along with the appropriate wound management, it is imperative for a dressing to facilitate the wound repair process too. In the present research, we hypothesize to improve the wound healing process by applying cost effective natural cocktail of various bioactive factors. Bovine colostrum contains high levels of immunoglobulins, lactoferrin, hormones and cytokines which play significant role in wound healing. Hence, multifunctional colostrum conjugated PCL-PEG based nanofibrous dressings were developed and analyzed for their physicochemical properties and cellular responses. The dressings were also evaluated for cell migration, antioxidant, anti-inflammatory and anti-bacterial properties. In-vivo wound healing ability was validated on a rat wound model. Numerous growth factors present in the colostrum showed their role in stimulation of skin repair and regeneration by direct action on genetic material. Significantly less inflammation in colostrum treated wounds was observed due to anti-inflammatory properties of lactoferrin. Thus obtained results confirmed the suitability of these multifunctional colostrum conjugated nanofibrous dressings for improved wound healing.

3D Bioprinted Alginate-Silk-Based Smart Cell-Instructive Scaffolds for Dual Differentiation of Human Mesenchymal Stem Cells.

Designing smart bioinks, which can provide multifunctionality and instructive cues to cells, is a current need of the tissue engineering field. Addressing these parameters, this work aims at developing a smart dual 3D bioprinted scaffold that is capable of differentiating human mesenchymal stem cells into two different lineages within the same construct without providing any exogenous cues. Here, biocompatible alginate- and silk-based bioinks were developed to print self-standing structures with the ability of spatially controlled differentiation of the encapsulated hMSCs. We present this proof of concept and have demonstrated a smart design where the incorporation of phosphate groups enhanced the osteogenic differentiation, whereas the addition of silk promoted the chondrogenic differentiation. Altogether, the present work suggests the potential of the developed bioinks for use in creating clinically viable osteochondral grafts.

Synergy-Based Motor Therapy Inducing Favorable Changes in Motor Function Components among Poststroke Subjects: A Single-Group Study.

Background Synergy is an outcome of multiple muscles acting in a synchronized pattern, controlled by the central nervous system. After brain insult, a set of deviated movement pattern emerges in the affected limb. The methods to train synchronization of muscles may diminish the deviated movement augmenting neuromotor control. The purpose of this investigation was to develop a synergy-based motor therapy (SBMT) protocol for the paretic upper limb in poststroke subjects. Further, the feasibility and effectiveness of the program was evaluated. . Methods The design was Pretest-posttest single-group assessor-blinded trial. Department of occupational therapy of a national institute for persons with physical disabilities was the study site. There were 40 study subjects (23 men, ranging from 40 to 60 years, 18 subjects with hemorrhagic cerebrovascular accident, and > 6 months after the accident) exhibiting motor paresis of half side of the body. SBMT is a stage-specific regime based on the linkage between the deviated and usual muscle action. SBMT items were selected considering the strength and magnitude of the deviated motor components. The movement linkages were utilized to dissociate strong coupled components; for instance, forearm pronation-supination with elbow 90-degree flexion. Fugl-Meyer Assessment (upper extremity) (FMA-UE), Wolf Motor Function Test (WMFT), and Barthel Index (BI) were applied to quantify the motor status, motor functional ability of the upper extremity, and self-care activities, respectively. Results All the enrolled subjects could perform their corresponding SBMT sessions. Posttreatment, FMA-UE improved significantly ( p < 0.001) from mean of 26.30 (standard deviation [SD] 15.02) to 35.20 (SD 17.64). Similarly, the WMFT both time (in seconds) and quality also positively improved significantly ( p < .001) from mean of 76.77 (SD 54.73) to 64.07 (SD 56.99) and 1.34 (SD 1.06) to 1.87 (SD 1.34), respectively. BI improved from 79.88 (SD 17.07) to 92.62 (SD 21.2) after the intervention ( p < 0.001). Conclusion SBMT protocol was a feasible and effective intervention to facilitate motor function components in chronic hemiparetic subjects. The regime could be considered as a potential intervention for stroke rehabilitation. Further trials and use of sophisticated measures are recommended to authenticate the outcome of this investigation. Clinical Trial Registration Clinical Trial Registry of India as CTRI/2017/10/010162 on October 23, 2017 (retrospectively).

Systematic development of extraction methods for quantitative microplastics analysis in soils using metal-doped plastics.

The inconsistency of available methods and the lack of harmonization in current microplastics (MPs) analysis in soils demand approaches for extraction and quantification which can be utilized across a wide variety of soil types. To enable robust and accurate assessment of extraction workflows, PET MPs with an inorganic tracer (Indium, 0.2% wt) were spiked into individual soil subgroups and standard soils with varying compositions. Due to the selectivity of the metal tracer, MPs recovery rates could be quickly and quantitatively assessed using ICP-MS. The evaluation of different methods specifically adapted to the soil properties were assessed by isolating MPs from complex soil matrices by systematically investigating specific subgroups (sand, silt, clay, non-lignified and lignified organic matter) before applying the workflow to standard soils. Removal of recalcitrant organic matter is one of the major hurdles in isolating MPs for further size and chemical characterization, requiring novel approaches to remove lignocellulosic structures. Therefore, a new biotechnological method (3-F-Ultra) was developed which mimics natural degradation processes occurring in aerobic (Fenton) and anaerobic fungi (CAZymes). Finally, a Nile Red staining protocol was developed to evaluate the suitability of the workflow for non-metal-doped MPs, which requires a filter with minimal background residues for further chemical identification, e.g. by µFTIR spectroscopy. Image analysis was performed using a Deep Learning tool, allowing for discrimination between the number of residues in bright-field and MPs counted in fluorescence mode to calculate a Filter Clearness Index (FCI). To validate the workflow, three well-characterized standard soils were analyzed applying the final method, with recoveries of 88% for MPs fragments and 74% for MPs fibers with an average FCI of 0.75. Collectively, this workflow improves our current understanding of how to adapt extraction protocols according to the target soil composition, allowing for improved MPs analysis in environmental sampling campaigns.