Craigfagan9477

Media and feed optimization have fueled many-fold improvements in mammalian biopharmaceutical production, but genome editing offers an emerging avenue for further enhancing cell metabolism and bioproduction. However, the complexity of metabolism, involving thousands of genes, makes it unclear which engineering strategies will result in desired traits. Here we present a comprehensive pooled CRISPR screen for CHO cell metabolism, including ~16,000 gRNAs against ~2500 metabolic enzymes and regulators. Using this screen, we identified a glutamine response network in CHO cells. BMS-232632 Glutamine is particularly important since it is often over-fed to drive increased TCA cycle flux, but toxic ammonia may accumulate. With the screen we found one orphan glutamine-responsive gene with no clear connection to our network. Knockout of this novel and poorly characterized lipase, Abhd11, substantially increased growth in glutamine-free media by altering the regulation of the TCA cycle. Thus, the screen provides an invaluable targeted platform to comprehensively study genes involved in any metabolic trait, and elucidate novel regulators of metabolism.In cell culture processes cell growth and metabolism drive changes in the chemical environment of the culture. These environmental changes elicit reactor control actions, cell growth response, and are sensed by cell signaling pathways that influence metabolism. link2 The interplay of these forces shapes the culture dynamics through different stages of cell cultivation and the outcome greatly affects process productivity, product quality, and robustness. Developing a systems model that describes the interactions of those major players in the cell culture system can lead to better process understanding and enhance process robustness. Here we report the construction of a hybrid mechanistic-empirical bioprocess model which integrates a mechanistic metabolic model with subcomponent models for cell growth, signaling regulation, and the bioreactor environment for in silico exploration of process scenarios. Model parameters were optimized by fitting to a dataset of cell culture manufacturing process which exhibits variabilctor scaling.

We present the Clinical Trial Knowledge Base, a regularly updated knowledge base of discrete clinical trial eligibility criteria equipped with a web-based user interface for querying and aggregate analysis of common eligibility criteria.

We used a natural language processing (NLP) tool named Criteria2Query (Yuan et al., 2019) to transform free text clinical trial eligibility criteria from ClinicalTrials.gov into discrete criteria concepts and attributes encoded using the widely adopted Observational Medical Outcomes Partnership (OMOP) Common Data Model (CDM) and stored in a relational SQL database. A web application accessible via RESTful APIs was implemented to enable queries and visual aggregate analyses. We demonstrate CTKB's potential role in EHR phenotype knowledge engineering using ten validated phenotyping algorithms.

At the time of writing, CTKB contained 87,504 distinctive OMOP CDM standard concepts, including Condition (47.82%), Drug (23.01%), Procedure (13.73%), Measurement (24.70%) and Observation (5.28%), with 34.78% for inclusion criteria and 65.22% for exclusion criteria, extracted from 352,110 clinical trials. The average hit rate of criteria concepts in eMERGE phenotype algorithms is 77.56%.

CTKB is a novel comprehensive knowledge base of discrete eligibility criteria concepts with the potential to enable knowledge engineering for clinical trial cohort definition, clinical trial population representativeness assessment, electronical phenotyping, and data gap analyses for using electronic health records to support clinical trial recruitment.

CTKB is a novel comprehensive knowledge base of discrete eligibility criteria concepts with the potential to enable knowledge engineering for clinical trial cohort definition, clinical trial population representativeness assessment, electronical phenotyping, and data gap analyses for using electronic health records to support clinical trial recruitment.Health information exchange (HIE) has mostly emerged as centralized data hubs that can pass data requests from one subscribing healthcare institution to another. Using traditional health information systems (HISs) with different technologies in hospitals leads to usability and incompatibility issues because of islands of information. This paper discusses shifting from HIE into an integrated universal health information infrastructure. Migration to such integrated universal electronic health records architecture could support real-time HIE and advanced modern big data analytics. However, there are various standards and technologies to facilitate HIS integration, a significant amount of efforts is still needed.

The long-term outcomes of patients following Gram-negative bacteraemia (GNB) are poorly understood. Here we describe a cohort of patients with GNB over a 2-year period and determine factors associated with late mortality (death between Days 31 and 365 after detection of bacteraemia).

This was a single-centre, retrospective, observational cohort study of 789 patients with confirmed Escherichia coli, Klebsiella spp. or Pseudomonas aeruginosa bacteraemia with a follow-up of 1 year. Multivariable survival analysis was used to determine risk factors for late mortality in patients who survived the initial 30-day period of infection.

Overall, 1-year all-cause mortality was 36.2%, with 18.1% of patients dying within 30 days and 18.1% of patients suffering late mortality. An adverse antimicrobial resistance profile [hazard ratio (HR)=1.095 per any additional antimicrobial category, 95% confidence interval (CI) 1.018-1.178; P=0.014] and infection with P. aeruginosa (HR=2.08, 95% CI 1.11-3.88; P=0.022) were independent predictors of late mortality. Other significant factors included Charlson comorbidity index and length of hospitalisation after the index blood culture.

Patients with GNB have a poor long-term prognosis. Risk factors for greater mortality at 1 year include co-morbidity, length of hospitalisation, and infecting organism and its resistance profile.

Patients with GNB have a poor long-term prognosis. Risk factors for greater mortality at 1 year include co-morbidity, length of hospitalisation, and infecting organism and its resistance profile.As the continuous opening-up and communication have been seen between China and other countries, traditional Chinese medical science and Chinese medicine now have gain popularity in China as well as the surrounding countries. As a window that China's opening-up through, Macau has been serving as a link between China and Portuguese language countries. The Forum for Economic and Trade Co-operation between China and Portuguese-speaking Countries (Macau), also known as Forum Macau, since it was established in 2003, has further enhanced the mutual exchange of commerce, culture and technologies. Promoting the industry of Chinese medical science among Portuguese-speaking countries has become one of the priorities of the Forum Macau. With multiple years of collaboration, among the Portuguese-speaking countries, Chinese medical science has gained promotion to some extent, as well as recognition from the government and people. The current study introduced the development of Chinese medical science among Portuguese-speaking countries from aspects such as legislation, cooperation and promotion, medical assistance, and clinical usage.Abnormal glycolytic metabolism contributes to angiogenic sprouting involved in atherogenesis. We investigated the potential anti-angiogenic properties of specific 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase-3 (PFKFB3) inhibitors in endothelial cells (ECs). ECs were treated with PFKFB3 inhibitors (named PA-1 and PA-2) and their effects on metabolic and functional characteristics of ECs were investigated. The anti-glycolytic compound 3-(pyridinyl)- 1-(4-pyridinyl)- 2-propen-1-one (3PO) was used as reference compound. PFKFB3 expression and activity (IC50 about 3-21 nM) was inhibited upon treatment with both compounds. Glucose uptake and lactate export were measured using commercial assays and showed a partial reduction up to 40%. PFKFB3 inhibition increased intracellular lactate accumulation, and reduced expression of monocarboxylate transporters-1 (MCT1) and MCT4. Furthermore, endothelial cell migration and proliferation assays demonstrated significant reduction upon treatment with both compounds. Matrix- metalloproteinase (MMP) activity, measured by gelatin zymography, and expression was significantly reduced (up to 25%). In addition, PA compounds downregulated the expression of VCAM-1, VE-cadherin, VEGFa, VEGFR2, TGF-β, and IL-1β, in inflamed ECs. Finally, PA-1 and PA-2 treatment impaired the formation of angiogenic sprouts measured by both morphogenesis and spheroid-based angiogenesis assays. Our data demonstrate that the anti-glycolytic PA compounds may affect several steps involved in angiogenesis. Targeting the key glycolytic enzyme PFKFB3 might represent an attractive therapeutic strategy to improve the efficacy of cancer treatments, or to be applied in other pathologies where angiogenesis is a detrimental factor.Metabolism of the essential amino acid tryptophan is a key metabolic pathway that restricts antitumor immunity and is a drug development target for cancer immunotherapy. Tryptophan metabolism is active in brain tumors including gliomas and promotes a malignant phenotype and contributes to the immunosuppressive tumor microenvironment. In recent years, improved understanding of the regulation and downstream function of tryptophan metabolism has been significantly expanded beyond the initial in vitro observation that the enzyme indoleamine-2,3-dioxygenase 1 (IDO1) promotes the depletion of intracellular tryptophan. Here, we revisit the specific roles of tryptophan metabolites in regulating brain functioning and neuronal integrity as well as in the context of brain tumors. This review summarizes recent developments in identifying key regulators, as well as the cellular and molecular effects of tryptophan metabolism with a particular focus on potential therapeutic targets in glioma.As the use of endovascular approaches to treat aneurysm repair continues to increase, more and more patients have been identified with endoleaks. Five types of endoleaks have been defined. Endotension, or type V endoleak, remains controversial owing to its variable definition across studies and the range of proposed treatments. link3 Thus, we performed a review of the reported studies to summarize the diagnosis and treatment of this rare complication after endovascular aneurysm repair to determine what we do and do not know about this rare form of endoleak. The presence of an endoleak places patients at an increased risk of aneurysm sac enlargement and potential rupture. Although additional research is essential and yet difficult to perform, we sought to provide a guide for the management of this perplexing endoleak known as endotension.

The Society for Vascular Surgery Wound, Ischemia, and foot Infection (WIfI) classification system was developed to stratify the risk of 1-year major amputation. Recently, the WIfI scores were used to define the estimated revascularization benefit quartiles ranging from high benefit (Q1) to questionable benefit (Q4). The aim of our study was to evaluate the revascularization benefit quartiles in a cohort of diabetic patients presenting with chronic limb-threatening ischemia (CLTI).

All diabetic patients presenting to our multidisciplinary diabetic foot and wound clinic (June 2012 to May 2020) who underwent lower extremity revascularization for CLTI were included. The affected limbs were graded using the WIfI system and assigned to an estimated benefit of revascularization quartile as previously published. One-year major amputation, complete foot healing, secondary patency, and amputation-free survival were calculated among the quartiles using Kaplan-Meier curve analyses and compared using Cox proportional hazards models.