Immobilization Protocol Research Articles

Effects of Skin Microclimate Changes on Skin Condition in Healthy Adults.

The purpose of this study was to examine the effects of moisture, as a skin microclimate variable, at the skin-support surface interface on repeated measures of skin erythema, stratum corneum hydration and skin temperature at pressure-prone areas of healthy adults. Quasi-experimental repeated measures study. Forty-eight healthy adults participated in a single center-controlled study conducted in a simulated hospital setting in an urban setting (Melbourne, Australia). This research project was conducted from November 2021 to June 2022. The effects of moisture and intermittent pressure-loading on skin overlying the sacrum, heels, and elbows were investigated. Participants followed a standardized immobilization protocol of 60minutes, with 10-minute brief pressure off-loading, in the semi-recumbent position on a standard hospital bed/mattress. The intervention comprised laying with the sacrum in contact with an absorbent pad saturated with normal saline; control skin sites (elbows and heels) were not exposed to the moistened absorbent pad. Skin measures were obtained consecutively at each anatomical testing site upon brief off-loading. Linear mixed models (LMMs) were used to compare skin parameters over time and between conditions. Differences in stratum corneum hydration scores between control and intervention conditions at the sacrum were significant (P =.001). Differences in erythema and skin temperature scores between control and intervention conditions at the sacrum did not significantly differ; however, significant changes in skin temperature (P =.025) at the sacrum were observed at the 10-minute measurement intervals over 1 hour in the sample, suggesting change in skin temperature over time. No participant displayed non-blanching erythema as a subjective indicator of skin injury in any of the testing conditions. Skin moisture/wetness, over a period of 1 hour, does not appear to impact erythema and skin temperature at the sacrum. In contrast, the introduction of moisture at the sacrum significantly increases sacral stratum corneum hydration. Study findings suggest that more frequent monitoring cycles of wet sacral skin may be required to identify moisture-associated skin changes.

Fabrication of a high-sensitivity electrochemical immunosensor by the oriented immobilization of engineered nanobody on nanofibrous membrane.

Anew type of label-free electrochemical immunosensor for the high-sensitivity determination of parathion was developed based on the oriented immobilization of nanobody (VHH9) on agold nanoparticle-loaded polyvinyl alcohol/citric acid nanofiber membrane-modified electrode. The morphology characterization and assembly process of the modified materials were investigated using scanning electron microscopy (SEM) and electrochemical impedance spectroscopy (EIS). Under the optimum conditions, the label-free electrochemical immunosensor for parathion exhibited a linear range of 0.0015-6400ng/mL and a low detection limit of 0.48pg/mL, the signal response of which was 10 times higher than that of the randomly immobilized VHH9. The immunosensor possessed high selectivity, good repeatability and reusability (keeping above 90% of its initial activity after repeating 8 times), and stability (remaining 90% after 9weeks of storage). Finally, the average recoveries of parathion from food samples were 93.76-105.73% with the coefficient of variation being 2.65-6.85%, showing good correlation with UPLC (R2 = 0.9950). Therefore, our nanobody immobilization protocol is simple and effective and proves the potential to be utilized as a promising candidate for sensing platform.

Removal of raffinose and stachyose from soy-based beverages using soluble and immobilized MelA α-D-galactosidase from Lactiplantibacillus plantarum WCFS1

The hydrolysis of α-galactose-carbohydrates in soy-based beverages using the enzyme MelA α-D-galactosidase (Lp_3485) from Lactiplantibacillus plantarum WCFS1 is presented. MelA is a highly unstable trimeric enzyme whose activity and stability are strongly dependent on the dilution and polarity of the reaction medium. Considering this, the enzyme was immobilized and stabilized using different immobilization protocols. Post-immobilization techniques consisting of cross-linking with different polyfunctional polymers were used. The optimal derivative was used to hydrolyse different oligosaccharides of the raffinose family (raffinose and stachyose) present in soy drink preparations. Remarkably, MelA showed specific activity and high hydrolytic efficiency towards RFOs in soy-based beverages at a relatively low temperature (20 °C). Therefore, these results suggest that this enzyme could be preferentially used in the early stage of soybean processing to remove RFOs from soy-based beverages and soy-derived products, thereby improving their nutritional value.

Does postoperative immobilization affect final alignment of pediatric femur fractures treated with flexible intramedullary nailing?

Flexible intramedullary nail (FIN) fixation of pediatric femur fractures is a popular method of fixation in children. Typical immobilization options include spica casting, long leg cast, knee immobilizers, or no immobilization and the decision to use each is usually left to surgeon preference. Our primary aim is to evaluate whether different postoperative immobilization status influences outcomes, namely radiographic alignment at the time of healing. A retrospective chart review was conducted of all patients with femur fractures treated with FIN fixation at a pediatric hospital from April 2018 through July 2022. Postoperative immobilization protocols were recorded and separated into two groups, patients who were immobilized (IMM) and patients who were not immobilized (NoIMM). Radiographs were evaluated for fracture alignment immediately following surgery and at the time of healing. Patients demographic, fracture, and postoperative clinical characteristics were compared between the two groups (NoIMM vs. IMM). A total of 41 patients were treated for diaphyseal femur fractures with FINs at our institution during the study period. No significant difference was observed in alignment at healing. Our results suggest that either immobilization or no immobilization after flexible intramedullary nailing of pediatric femoral shaft fractures are viable options when postoperative immobilization status is left to the surgeon's discretion.

Microtiter Plate Immobilization Screening for Prototyping Heterogeneous Enzyme Cascades.

Immobilization is a key enabling technology in applied biocatalysis that facilitates the separation, recovery, and reuse of heterogeneous biocatalysts. However, finding a consensus immobilization protocol for several enzymes forming a multi-enzyme system is extremely difficult and relies on a combinatorial trial-and-error approach. Herein, we describe a protocol in which 17 different carriers functionalized with different reactive groups are tested in a 96-well microtiter plate to screen up to 21 immobilization protocols for up to 18 enzymes. This screening includes an activity and stability assay to select the optimal immobilization chemistry to achieve the most active and stable heterogeneous biocatalysts. The information retrieved from the screening can be rationalized using a Python-based application CapiPy. Finally, through scoring the screening results, we find the consensus immobilization protocol to assemble an immobilized four-enzyme system to transform vinyl acetate into (S)-3-hydroxybutyric acid. This methodology opens a path to speed up the prototyping of immobilized multi-enzyme pathways for chemical manufacturing.

Microtiter Plate Immobilization Screening for Prototyping Heterogeneous Enzyme Cascades

AbstractImmobilization is a key enabling technology in applied biocatalysis that facilitates the separation, recovery, and reuse of heterogeneous biocatalysts. However, finding a consensus immobilization protocol for several enzymes forming a multi‐enzyme system is extremely difficult and relies on a combinatorial trial‐and‐error approach. Herein, we describe a protocol in which 17 different carriers functionalized with different reactive groups are tested in a 96‐well microtiter plate to screen up to 21 immobilization protocols for up to 18 enzymes. This screening includes an activity and stability assay to select the optimal immobilization chemistry to achieve the most active and stable heterogeneous biocatalysts. The information retrieved from the screening can be rationalized using a Python‐based application CapiPy. Finally, through scoring the screening results, we find the consensus immobilization protocol to assemble an immobilized four‐enzyme system to transform vinyl acetate into (S)‐3‐hydroxybutyric acid. This methodology opens a path to speed up the prototyping of immobilized multi‐enzyme pathways for chemical manufacturing.

Enzyme-Immobilized Porous Crystals for Environmental Applications.

Developing efficient technologies to eliminate or degrade contaminants is paramount for environmental protection. Biocatalytic decontamination offers distinct advantages in terms of selectivity and efficiency; however, it still remains challenging when applied in complex environmental matrices. The main challenge originates from the instability and difficult-to-separate attributes of fragile enzymes, which also results in issues of compromised activity, poor reusability, low cost-effectiveness, etc. One viable solution to harness biocatalysis in complex environments is known as enzyme immobilization, where a flexible enzyme is tightly fixed in a solid carrier. In the case where a reticular crystal is utilized as the support, it is feasible to engineer next-generation biohybrid catalysts functional in complicated environmental media. This can be interpreted by three aspects: (1) the highly crystalline skeleton can shield the immobilized enzyme against external stressors. (2) The porous network ensures the high accessibility of the interior enzyme for catalytic decontamination. And (3) the adjustable and unambiguous structure of the reticular framework favors in-depth understanding of the interfacial interaction between the framework and enzyme, which can in turn guide us in designing highly active biocomposites. This Review aims to introduce this emerging biocatalysis technology for environmental decontamination involving pollutant degradation and greenhouse gas (carbon dioxide) conversion, with emphasis on the enzyme immobilization protocols and diverse catalysis principles including single enzyme catalysis, catalysis involving enzyme cascades, and photoenzyme-coupled catalysis. Additionally, the remaining challenges and forward-looking directions in this field are discussed. We believe that this Review may offer a useful biocatalytic technology to contribute to environmental decontamination in a green and sustainable manner and will inspire more researchers at the intersection of the environment science, biochemistry, and materials science communities to co-solve environmental problems.

Development of Spiropyran Immobilization and Characterization Protocols for Reversible Photopatterning of SiO2 Surfaces.

Spiropyran is a dynamic organic compound that is distinguished by its reversible conversion between two forms: the colorless closed spiropyran (SP) form and the purple open merocyanine (MC) form. Typically triggered by UV light and reversed by visible light, spiropyran-functionalized surfaces offer reversible conversion in properties including color, polarity, reactivity, and fluorescence, making them applicable to diverse applications in chemical sensors, biosensors, drug delivery, and heavy metal extraction. While spiropyran has been successfully incorporated into various material platforms with SiO2 surfaces, its application on flat surfaces has been limited due to surface area constraints and a lack of standardized evaluation methods, which largely depend on the integration approach and substrate type used. In this study, we systematically review the existing literature and categorize integration methods and substrate types first and then report on our experimental work, in which we developed a streamlined three-step immobilization protocol, which includes surface activation, amination with (3-aminopropyl) triethoxysilane (APTES), and subsequent functionalization with carboxylic spiropyran (SP-COOH). Using SiO2 surfaces as a demonstration, we have also established a robust characterization protocol, consisting of contact angle measurements, X-ray photoelectron spectroscopy (XPS), ellipsometry, and fluorometric analysis. Our results evaluate the newly developed immobilization protocol, demonstrating effective activation and optimal amination using a 2% APTES solution, achieved in 5 min at room temperature. Fluorescence imaging provided clear contrast between the SP and the MC forms. Furthermore, we discuss limitations in the surface density of functional groups and steric hindrance and propose future improvements. Our work not only underscores the versatility of spiropyran in surface patterning but also provides optimized protocols for its immobilization and characterization on SiO2 surfaces, which may be adapted for use on other substrates. These advancements lay the groundwork for on-chip sensing technologies and other applications.

A universal method for surface-based binding assays by preparing immobilized β2-adrenergic receptor stationary phase using solid binding peptide as a fusion tag

Protein functionalized surface has the potential to develop new assays for determining the drug-like properties of potential compounds and discovering specific partners of G protein-coupled receptors (GPCRs). However, a universal method for purifying and immobilizing functional GPCRs has remained elusive. To this end, we developed a general and rapid way to purify and immobilize β2-adrenergic receptor (β2AR) by silicon-specific peptide. We screened CotB1p as a tag from six silica-binding peptides (minTBP-1, CotB1p, SB7, Car9, and Si4–1) by examining their affinity to macroporous silica gel. We investigated the adsorption and desorption of CotB1p-tagged β2-adrenoceptor (β2AR-CotB1p) under diverse conditions to propose a protocol for receptor purification and immobilization. Under optimized conditions, β2AR immobilization were achieved by directly immersing cell lysates harboring the receptor with silica gel, and the elution of the receptor without demonstratable contaminants was realized by including l-arginine/L-lysine in the elutes. This allows purification of the receptor from Escherichia coli (E.coli) lysates with a purity of 95 %. The immobilized receptor was utilized as a stationary phase to reveal the tag impact on ligand-binding outputs by comparing the CotB1p-strategy with a typical covalent method. The KAs of salbutamol, chlorprenaline, tulobuterol, and terbutaline on β2AR-CotB1p column were 1.26 × 106, 6.59 × 106, 7.90 × 106, and 8.97 × 105 M−1 respectively, which were two orders of magnitude higher than those on the Halo-β2AR column. The whole immobilization was accomplished within 30 min without the requirement of any special treatment, resulting in enhanced accuracy for determining receptor-ligand binding parameters. Taken together, CotB1p-mediated strategy is simple, rapid, and universal for purification or immobilization of unstable biomolecules like GPCRs for analytical and biological applications.

Evaluation of Proteolytic Digestion Efficiency in Hydrogen Exchange-Mass Spectrometry Experiments Using the Digestible Peptide Score.

In a hydrogen exchange-mass spectrometry (HX-MS) experiment, the enzymatic proteolysis of the deuterated protein is an essential step. Often the differences in the performance between different digestion protocols or between immobilized protease columns can be challenging to evaluate. To compare differences in the performance of immobilized protease columns, a new digestion efficiency metric known as digestible peptide scoring (DPS) was developed and is presented in this work. The measured response fraction of substance P peptide is used to assign a value between 0% and 100% based on the fraction of substance P digested by the enzyme, using angiotensin II as an undigested internal standard. In this work, the DPS approach was tested using multiple immobilized pepsin batches prepared using different protocols. The results demonstrate the repeatability of DPS values for batches prepared using the same conditions and the ability of the DPS evaluations to provide unique values when the immobilization conditions were altered. Protein digestions obtained with a higher scoring column were better than digestions obtained using a lower scoring column. The DPS evaluation is simple and quickly provides an unambiguous assessment which can be used to evaluate an immobilized enzyme column's suitability prior to performing an experiment, to track performance over a column's lifetime, to optimize protease immobilization protocols specifically for the quench conditions of a particular experiment, and to optimize the digestion conditions.

Neuromuscular Adaptations in Response to Disuse and Weight-Training Rehabilitation are Enhanced in Resistance Trained Individuals

INTRODUCTION: Lower-limb disuse from immobilization results in a decrease in muscle strength, volume, and function. However, muscle atrophy from disuse does not entirely account for the observed strength and functional declines. Additionally, prior engagement in resistance training (RT) has been linked to the preservation of muscular strength and neuromuscular function. Therefore, the purpose of this study was to assess the influence of a single-leg immobilization protocol, followed by structured RT rehabilitation protocol, on muscle strength, volume, and neural activation in healthy young adults with and without previous strength training experience. Methods: Participants (10 Trained (T): 8 men, 2 women; 11 Untrained (UT): 8 men, 3 women) underwent 2-weeks of left-knee immobilization at 90° knee-flexion with a hinged immobilizer knee-brace. Participants were given a pair of axillary crutches for ambulation during this time. Following immobilization, all participants underwent an 8-week structured RT rehabilitation protocol. Participants visited the laboratory for three testing sessions (baseline test on Day 0; post-immobilization at the end of week 2; and post-RT rehab at the end of week 10). Maximal knee extensor isometric strength at 45°was quantified using an isokinetic dynamometer (Biodex). Voluntary muscle activation was quantified as the root mean square (RMS) of the interference electromyography (EMG) signals derived from two indwelling fine-wire electrodes in the vastus-lateralis muscle. Additionally, a continuous wavelet transform (CWT) was performed on the EMG to determine amplitude versus frequency characteristics of the signals and how this amplitude varied with time in the 13-30 (beta) and 30-60 (piper) Hz bands. Mid-thigh muscle cross-sectional area (mCSA) and density were also quantified using peripheral quantitative computed tomography (pQCT) during all testing sessions. Results: The T group was significantly stronger (p=0.019), had greater muscle density (p=0.005) and mid-thigh mCSA (p<0.001) across all sessions when compared with the UT group. However, there were no significant time or interaction effects for strength or muscle density losses (p>0.05). While both groups demonstrated increased muscle activation across sessions (RMS: p=0.04; CWT: p=0.021), the T group demonstrated a greater increase in the neural drive from baseline to week 10 when compared with the UT group (RMS: p= 0.002; CWT: p=0.027). CONCLUSION: In younger adults with or without prior strength training experience, we find that losses in knee extensor isometric strength, muscle density, and mid-thigh mCSA are minimal following a 2-week immobilization protocol. However, certain neuromuscular adaptations during immobilization and following weight-training rehabilitation are enhanced to a greater degree in those with prior training, which is seemingly indicative of greater synchrony in descending cortical drive and motor unit recruitment. These latter findings warrant continued investigation using additional innovative methodologies (e.g., fMRI, TMS, etc.) for assessing the mechanisms underlying the effects of prehab protocols on neuromuscular adaptations. PGRF and Nix family foundation scholarship to Marina Meyer. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Living emission abolish filters (LEAFs) for methane mitigation: design and operation

As one of the most potent greenhouse gases, methane is a critical target for the near-term mitigation of global warming. Efficient, scalable, easy-to-implement, and robust mitigation technologies are urgently needed to assist in reaching methane abolishment. The goal of this research was to test the applicability of active, extremophilic methanotrophic cells as a baseline concept for engineered systems aiming at methane capturing. The system, named living emission abolish filters (LEAFs), represents an array of immobilized biomaterials capable of capturing methane directly from vent streams. The biomaterials were made using cells of Methylotuvimicrobium alcaliphilum 20ZR, a robust halophilic methanotrophic bacterium with the ability to consume methane gas at low concentrations. Several critical parameters were tested, including (i) the composition of the matrix and optimal immobilization to increase catalyst load, (ii) the stability of methanotrophic cells, and (iii) the toxicity of trace gases (i.e. CO). We found that hydrogels coated with 2.3 mg cell dry weight/cm3 methanotrophic cells represent the best-performing biomaterials. The methane reduction potential of LEAFs fluctuated from 20% to 95% and depended on the methane concentration in the gas stream and the stream flow rates. The potential for commercial-scale deployment and emissions reductions was also evaluated. Total greenhouse gas emissions (combined using the global warming potential GWP100) from an example using a ventilation air methane source over a one-year period was shown to be reduced in two LEAF scenarios by 51% and 75%. Over longer time horizons, more significant reductions are possible as consistent methane consumption can be sustained. The study highlights the overall potential of the liquid-free bio-based composite methane mitigation system. Further improvements essential for system assembly and implementations should include (a) optimization of the cell immobilization protocols to improve cell load and the shelf-life of the system and (b) implementation of matrix moldings for cell immobilization to achieve optimal gas flow and increase the cell-gas interface.

Proteases: Importance, Immobilization Protocols, Potential of Activated Carbon as Support, and the Importance of Modifying Supports for Immobilization.

Although enzymes have been used for thousands of years, their application in industrial processes has gained importance since the 20th century due to technological and scientific advances in several areas, including biochemistry [...].

Development and testing of a sedation protocol for Neocaridina davidi

Neocaridina davidi, a small freshwater shrimp native to Asia, specifically China, Japan, Korea, and Vietnam, possesses remarkable resistance to poor water quality and offers various advantages over other invertebrate species to examine crucial issues in neuroscience and other related areas. These advantages include robustness, ease of maintenance, and transparency, making them useful for in vivo studies with optical imaging techniques. Despite its suitability for research purposes, particularly in the fields of imaging and fluorescent techniques, the lack of attention given to this species has resulted in the absence of a robust and replicable sedation protocol for immobilization and safe manipulation. Consequently, researchers face challenges in performing experimental procedures while minimizing harm to this specimen. In this study, we have developed and evaluated a simple sedation protocol specifically designed for Neocaridina davidi, assessing its effectiveness using light microscopy and image processing.

Clinical outcomes after primary repair for thumb ulnar collateral ligament ruptures: a systematic review and meta-analysis.

There is no consensus on the best surgical technique for treating thumb ulnar collateral ligament (UCL) ruptures. This systematic review and meta-analysis investigates which primary repair technique and postoperative immobilization protocol result in the best clinical outcomes. A literature search was conducted in Embase, Medline ALL Ovid, Web of Science Core Selection and Cochrane Central Register of Controlled Trials. Pain, stability, tip pinch strength, key pinch strength, grip strength, return to work and metacarpophalangeal joint range of motion were collected as postoperative outcomes. In total, 29 studies were included. All surgical techniques resulted in satisfactory clinical outcomes, with no significant differences between bone anchor reinsertion, suture fixation, K-wire fixation and a combination of techniques. K-wire immobilization resulted in worse postoperative pain, but similar stability compared to immobilization without a K-wire. Clinical outcomes after thumb UCL repair are excellent, with no differences in clinical outcomes noted among surgical techniques.

Improving the stability of an unstable lipase by applying different immobilization strategies for the selective hydrolysis of fish oil

AbstractRhizopus oryzae lipase (ROL) is known to present high selectivity in chemical reactions. However, the poor stability of ROL effectively limits its industrial applications. In this study, several immobilization protocols, such as hydrophobic adsorption, covalent immobilization, multi‐point covalent attachment, ionic adsorption/cross‐linking, and ionic interaction, were applied to improve the stability of ROL. Heterogeneous modification of aspartic and glutamic acid residues on the surface of ROL was carried out by 1‐ethyl‐3‐(3‐dimethylaminopropyl) carbodiimide (EDC) to introduce new amine groups with lower pKb. The highest immobilization yield of 89% was achieved for octyl‐agarose, producing specific activity of 45 U/mg, which is 15 folds higher than the specific activity of the soluble enzyme. Improved stability of ROL was observed, in particular for those derivatives obtained by multi‐point covalent attachment of ROL on glyoxyl‐agarose (Gx‐ROL) and aminated ROL on glyoxyl‐agarose (Gx‐NH2‐ROL) by retaining 28%–36% of their initial activities after 24 h incubation at 60°C. Immobilization also altered the co‐solvent stability profile of the immobilized derivatives producing biocatalysts with varied co‐solvent stabilities. Furthermore, utilization of the immobilized preparations in fish oil hydrolysis revealed the selective release of cis‐5,8,11,14,17‐eicosapentaenoic acid (EPA) and cis‐4,7,10,13,16,19‐docosahexaenoic acid (DHA) in favor of EPA. The highest EPA/DHA selectivity of 33 was observed for the hydrophobically immobilized ROL on octyl‐sepharose.

Development and Evaluation of an Expedited System for Creation of Single Walled Carbon Nanotube Platforms.

Single-walled carbon nanotubes (SWNT) have a strong and stable near-infrared (nIR) fluorescence that can be used to selectively detect target analytes, even at the single molecule level, through changes in either their fluorescence intensity or emission peak wavelength. SWNTs have been employed as NIR optical sensors for detecting a variety of analytes. However, high costs, long fabrication times, and poor distributions limit the current methods for immobilizing SWNT sensors on solid substrates. Recently, our group reported a protocol for SWNT immobilization with high fluorescence yield, longevity, fluorescence distribution, and sensor response, unfortunately this process takes 5 days to complete. Herein we report an improved method to immobilize SWNT sensors that only takes 2 days and results in higher fluorescence intensity while maintaining a high level of SWNT distribution. We performed surface morphology and chemical composition tests on the original and new synthesis methods and compared the sensor response rates. The development of this new method of attaching SWNT sensors to a platform allows for creation of a sensing system in just 2 days without sacrificing the advantageous characteristics of the original, 5-day platforms.

Advancements in enzyme immobilization on magnetic nanomaterials: toward sustainable industrial applications.

Enzymes are widely used in biofuels, food, and pharmaceuticals. The immobilization of enzymes on solid supports, particularly magnetic nanomaterials, enhances their stability and catalytic activity. Magnetic nanomaterials are chosen for their versatility, large surface area, and superparamagnetic properties, which allow for easy separation and reuse in industrial processes. Researchers focus on the synthesis of appropriate nanomaterials tailored for specific purposes. Immobilization protocols are predefined and adapted to both enzymes and support requirements for optimal efficiency. This review provides a detailed exploration of the application of magnetic nanomaterials in enzyme immobilization protocols. It covers methods, challenges, advantages, and future perspectives, starting with general aspects of magnetic nanomaterials, their synthesis, and applications as matrices for solid enzyme stabilization. The discussion then delves into existing enzymatic immobilization methods on magnetic nanomaterials, highlighting advantages, challenges, and potential applications. Further sections explore the industrial use of various enzymes immobilized on these materials, the development of enzyme-based bioreactors, and prospects for these biocatalysts. In summary, this review provides a concise comparison of the use of magnetic nanomaterials for enzyme stabilization, highlighting potential industrial applications and contributing to manufacturing optimization.

An Expedited Sling Immobilization Protocol After Isolated Biceps Tenodesis Results in Clinical and Patient-Reported Postoperative Outcomes Equivalent to a Standard Rehabilitation Protocol

PurposeTo characterize clinical and patient-reported outcomes for patients after isolated biceps tenodesis (BT) who underwent either standard or expedited sling immobilization protocols following surgery. MethodsThis retrospective cohort study comparing patients who were assigned to use a sling for either four- to six-weeks (standard) or zero- to two-weeks (expedited) following an isolated BT. Primary endpoint included rate of re-rupture, surgical revision, loss of fixation, and Popeye deformity. Secondary endpoints included shoulder range of motion (ROM) as well as pre- and post-operative patient reported outcomes (PROs) of pain and function. Missing data was managed via multiple imputation with chained equations. Complication prevalence 95% confidence intervals were calculated using the Clopper Pearson method and a series of hierarchical mixed effects linear regressions were performed to assess differences between sling interventions in PROs and ROM. ResultsThe average age of the standard cohort (n=66) was 49 years ( ± 14) and the average age of the expedited cohort (n=69) was 47 years (± 14). The expedited and standardized cohorts demonstrated 0.4 and 0.3 complications per 10,000 exposure days, respectively, with no significant difference between groups [1.4 (95% CI: 0.2, 10.0), p = 0.727]. There was no demonstrated difference in forward flexion, abduction, or external range of motion. The expedited group had less improvement in Visual Analogue Scale (VAS) for pain scores that was not clinically significant and there were no differences in PROs of function. ConclusionsNo statistically significant difference in the rate of re-rupture, surgical revision, loss-of-fixation, or Popeye deformity was noted between protocols after isolated BT. Furthermore, there were no clinically significant differences in ROM or PROs identified between protocols after isolated BT. This study suggests that patients who have undergone isolated BT may safely discontinue sling use within two weeks after surgery.

Network‐based systematic dissection of exercise‐induced inhibition of myosteatosis in older individuals

AbstractAccumulated fat in skeletal muscle (i.e. myosteatosis), common in sedentary older individuals, compromises skeletal muscle health and function. A mechanistic understanding of how physical activity levels dictate fat accumulation represents a critical step towards establishment of therapies that promote healthy ageing. Using a network medicine paradigm that characterized the transcriptomic response of aged muscle to exercise versus immobilization protocols, this study explored the shared molecular cascade that regulates the fate of fibro‐adipogenic progenitors (FAPs), the cell population primarily responsible for fat accumulation. Specifically, gene set enrichment analyses with network propagation revealed Pgc‐1α as a functional hub of a large gene regulatory network underlying the regulation of FAPs by physical activity in aged muscle, but not in young counterparts. Integrated in silico and in situ approaches to induce Pgc‐1α overexpression in aged muscle promoted mitochondrial fatty acid oxidation and inhibited FAP adipogenesis. These findings suggest that the Pgc‐1α–mitochondrial fatty acid oxidation axis is a shared mechanism by which physical activity regulates age‐related myosteatosis. The network medicine paradigm introduced provides mechanistic insight into exercise adaptation in elderly skeletal muscle and offers translational opportunities to advance exercise prescription for older populations. imageKey points Fat accumulation is a quintessential feature of aged skeletal muscle. While increasing physical activity levels has been proposed as an effective strategy to reduce the fat in skeletal muscle (i.e. myosteatosis), the molecular cascade underlying these benefits has been poorly defined. This study implemented a series of network medicine approaches and uncovered Pgc‐1α as a mechanistic driver of the regulation of fibro‐adipogenic progenitors (FAPs) by physical activity. Integrated in silico and in situ approaches to induce Pgc‐1α overexpression promoted mitochondrial fatty acid oxidation and inhibited FAP adipogenesis. Together, the findings of the current study suggest a novel hypothesis that physical activity reduces myosteatosis via upregulation of Pgc‐1α‐mediated mitochondrial fatty acid oxidation and subsequent inhibition of FAP adipogenesis.