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•Lyoprotectant combinations enhance lyophilized extract viability under 50 °C storage.•Lyoprotected single-pot cell-free protein synthesis (CFPS) retains >45% activity after 90-day ...ambient storage.•Lyoprotected single-pot CFPS systems maintain viability for 46 days at 50 °C.•Active, endotoxin-free crisantaspase is produced from stored CFRxlyo systems.
Cell-free protein synthesis has emerged as a promising platform for the production of therapeutic proteins due to its inherently open reaction environment, flexible reaction conditions and rapid protein synthesis capabilities. In recent years, lyophilized cell-free systems have widened the application space of cell-free technology by improving reagent stability outside of cold-chain storage. Current embodiments of the system, however, demonstrate poor stability at elevated temperatures. Lyoprotectants have long been recognized for the ability to preserve the activity of biological molecules during drying processes, but the application of this technology to lyophilized cell-free systems has been limited and has failed to address the negative effects that such lyoprotectants may have on cell-free systems. Here, several lyoprotected, lyophilized cell-free protein synthesis systems are demonstrated using antiplasticized sugar glasses as lyoprotectants, showing significant improvement over standard lyophilized systems or trehalose-preserved systems. Furthermore, we demonstrate for the first time, preservation and therapeutic expression, specifically of FDA-approved crisantaspase, from a truly single-pot lyophilized, endotoxin-free, cell-free protein synthesis system, exemplifying the potential for on-site therapeutic synthesis.
Automatic representation learning of key entities in electronic health record (EHR) data is a critical step for healthcare data mining that turns heterogeneous medical records into structured and ...actionable information. Here we propose ME2Vec, an algorithmic framework for learning continuous low-dimensional embedding vectors of the most common entities in EHR: medical services, doctors, and patients. ME2Vec features a hierarchical structure that encapsulates different node embedding schemes to cater for the unique characteristic of each medical entity. To embed medical services, we employ a biased-random-walk-based node embedding that leverages the irregular time intervals of medical services in EHR to embody their relative importance. To embed doctors and patients, we adhere to the principle "it's what you do that defines you" and derive their embeddings based on their interactions with other types of entities through graph neural network and proximity-preserving network embedding, respectively. Using real-world clinical data, we demonstrate the efficacy of ME2Vec over competitive baselines on diagnosis prediction, readmission prediction, as well as recommending doctors to patients based on their medical conditions. In addition, medical service embeddings pretrained using ME2Vec can substantially improve the performance of sequential models in predicting patients clinical outcomes. Overall, ME2Vec can serve as a general-purpose representation learning algorithm for EHR data and benefit various downstream tasks in terms of both performance and interpretability.
The flexible, electropositive cavity of linear 1,4-diaryl-1,2,3-triazole oligomers provides a suitable host for complexation of various anions. The binding affinities for various combinations of ...oligomer and anion were determined by (1)H NMR titrations. Effective ionic radius is found to be a primary determinant of the relative binding interactions of various guests, with small but measurable deviations in the case of nonspherical anions. Solvent effects are significant, and the strength of the binding interaction is found to depend directly on the donor ability of the solvent. A picture emerges in which anion binding can be effectively interpreted in terms of a competition between two solvation spheres: one provided by the solvent and a second dominated by a folded cavity lined with electropositive 1,2,3-triazole CH protons. Implications for rigid macrocycles and other multivalent hosts are discussed.
Objective: Exercise is beneficial for both the body and the mind, and it has been associated with protective neurocognitive effects, such as increased levels of brain-derived neurotrophic factor and ...neurogenesis. These effects are linked to the attenuation of age-related mental decline and the preservation of mental capacities in older, physically active adults. This study evaluated whether masters athletes, a highly active population, have better cognitive function compared to age-matched non-athletes based on the Immediate Post-concussion Assessment and Cognitive Testing (ImPACT) tool. Methods: Masters athletes and sedentary controls were recruited and screened for eligibility. All subjects were excluded if they had preexisting neurological diseases, psychiatric disorders, substance abuse disorders, learning disorders, and/or a history of traumatic brain injury, and in addition, control subjects were excluded if they performed >1 h/week of aerobic exercise. All participants completed a health and activity survey which includes the SF-12 and the ImPACT neurocognitive test which measures verbal and visual memory as well as reaction time. Differences between masters athletes and the control population were determined by ImPACT score composites. Results: 51 pairs of athletes and non-athletes were analyzed. Athletes had significantly higher verbal memory scores (85.9 ± 7.7 vs 79.9 ± 13.9, p = 0.01) and faster reaction times (0.71 ± 0.12 vs 0.76 ± 0.15 s, p = 0.04) on the ImPACT test. Athletes also scored significantly higher on the physical components summary score of the SF-12 (55.0 ± 3.3 vs 51.8 ± 6.7, p = 0.004). Conclusion: Masters athletes performed better on verbal memory and reaction time test, as well as on physical function as evaluated by the SF-12, compared to non-athlete controls. Chronic physical activity may preserve neurocognitive processes and increase physical health, which are protective factors for the negative effects of the aging process.
Estrogen, a naturally occurring female steroid growth hormone, has been implicated as a major risk factor for the development of breast cancer. Recent research into this disease has also correlated ...Annexin-1 (ANXA1), a glucocorticoid-inducible protein, with the development of breast tumorigenesis. ANXA1 is lost in many cancers, including breast cancer, and this may result in a functional promotion of tumor growth. In this study, we investigated the expression of ANXA1 in MCF-7 cells treated with estrogen and the regulation of estrogen functions by ANXA1. Exposure of MCF-7 breast cancer cells to high physiologic levels (up to 100 nmol/L) of estrogen leads to an up-regulation of ANXA1 expression partially through the activation of cyclic AMP-responsive element binding protein and dependency on activation of the estrogen receptor. In addition, treatment of MCF-7 cells with physiologic levels of estrogen (1 nmol/L) induced proliferation, whereas high pregnancy levels of estrogen (100 nmol/L) induced a growth arrest of MCF-7 cells, associated with constitutive activation of extracellular signal-regulated kinase 1/2 and up-regulation of cell cycle arrest proteins such as p21(waf/cip). Silencing of ANXA1 with specific small interfering RNA reverses the estrogen-dependent proliferation as well as growth arrest and concomitantly modulates extracellular signal-regulated kinase 1/2 phosphorylation. We confirm that ANXA1 is lost in clinical breast cancer, indicating that the antiproliferative protective function of ANXA1 against high levels of estrogen may be lost. Finally, we show that ANXA1-deficient mice exhibit faster carcinogen-induced tumor growth. Our data suggest that ANXA1 may act as a tumor suppressor gene and modulate the proliferative functions of estrogens.
Purpose of review
The use of biologics in orthopedics is becoming increasingly popular as an adjuvant in healing musculoskeletal injuries. Though many biologics involved in the management of foot and ...ankle injuries are used based on physician preference, reports of improved outcomes when combined with standard operative treatment has led to further clinical interest especially in foot and ankle trauma.
Recent findings
The most recent studies have shown benefits for biologic use in patients predisposed to poor bone and soft tissue healing. Biologics have shown benefit in treating soft tissue injuries such as Achilles ruptures as well as the complications of trauma such as non-unions and osteoarthritis.
Summary
Biologics have shown some benefit in improving functional and pain scores, as well as reducing time to heal in foot and ankle traumatic injuries, with particular success shown with patients that have risk factors for poor healing. As the use of biologics continues to increase, there is a need for high-level studies to confirm early findings of lower level reports.
...2016 marked the fifth consecutive year in which investment in renewables outstripped investment in fossil fuel technologies. The Trump administration, in contrast, has opened the Arctic National ...Wildlife Reserve (ANWR) and offshore waters for drilling, shrunk national monuments, proposed to repeal the current Clean Power Plan, explored the possibility of reopening considerations for fuel economy standards, considered new tariffs that would increase the cost of solar panels in the United States, dropped climate threats from the National Security Strategy, and, most infamously, announced its intention to withdraw from the Paris Climate Agreement. A 2015 review by the Obama Administration found that the fossil fuel industry received US$4.6 billion per year in specialized subsidies, without which a stunning 45 percent of oil production would not be profitable. Despite the fad of the government telling individuals how to combat climate change, impactful individual consumption is not possible within legislative, economic, and physical systems still based on fossil fuel.
An “endotoxin-free” E. coli-based cell-free protein synthesis system has been reported to produce therapeutic proteins rapidly and on-demand. However, preparation of the most complex CFPS reagent – ...the cell extract – remains time-consuming and labor-intensive because of the relatively slow growth kinetics of the endotoxin-free ClearColiTMBL21(DE3) strain. Here we report a streamlined procedure for preparing E. coli cell extract from ClearColi™ using auto-induction media. In this work, the term auto-induction describes cell culture media which eliminates the need for manual induction of protein expression. Culturing Clearcoli™ cells in autoinduction media significantly reduces the hands-on time required during extract preparation, and the resulting “endotoxin-free” cell extract maintained the same cell-free protein synthesis capability as extract produced with traditional induction as demonstrated by the high-yield expression of crisantaspase, an FDA approved leukemia therapeutic. It is anticipated that this work will lower the barrier for researchers to enter the field and use this technology as the method to produce endotoxin-free E. coli-based extract for CFPS.
Objective: To investigate the impact of loneliness on all-cause mortality in the oldest old population over a 10-year follow-up.
Method: Participants were from the third wave of the Cambridge City ...over-75s Cohort (CC75C) study, a population-based longitudinal study of older people aged 75 or over. Loneliness was measured two further times. At each wave, participants were asked how often they felt lonely and the answers were divided into three levels: not lonely, slightly lonely and lonely. The relationship between loneliness and all-cause mortality was examined using Cox regression with loneliness as a time-varying predictor. The association was adjusted for socio-demographic factors, number of chronic diseases, functional ability and depression.
Results: Seven hundred thirteen participants were seen at wave 3 (out of 2166 at baseline), of whom 665 had data on loneliness. The prevalence of feeling slightly lonely and lonely was 16% and 25%, respectively. Vital status was followed for a further 10 years. A total of 562 participants died during the follow-up. After adjusting for age, sex and other socio-demographic factors, loneliness was associated with a 20% increased risk of mortality (HR: 1.2, 95% CI: 1.0-1.6). The association was disappeared after further adjusting for health conditions and depression (HR: 1.0, 95% CI: 0.8-1.4). Individuals who reported being slightly lonely were not at risk of mortality.
Conclusions: The association between loneliness and mortality was fully explained by health conditions, suggesting that in the very old age, health problem is the proximal risk factor for mortality.
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