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Promoting the production of challenging proteins via induced expression in CHO cells and modified cell-free lysates harboring T7 RNA polymerase and mutant eIF2α

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Synthetic and Systems Biotechnology 2024年第3期9卷 416-424页

作者： Jeffrey L.Schloßhauer Lena Tholen Alexander Korner Stefan Kubick Sofia Chatzopoulou Anja Honow anne Zemella Fraunhofer Project Group PZ-Syn of the Fraunhofer Institute for Cell Therapy and Immunology-IZI Branch Bioanalytics and Bioprocesses-IZI-BBAm MühlenbergPotsdamGermany Fraunhofer Institute for Cell Therapy and Immunology-IZI Branch Bioanalytics and Bioprocesses-IZI-BBAm MühlenbergPotsdamGermany Faculty of Health Sciences Joint Faculty of the Brandenburg University of Technology Cottbus-SenftenbergThe Brandenburg Medical School Theodor FontaneUniversity of PotsdamPotsdamGermany Institute for Chemistry and Biochemistry Laboratory of Protein BiochemistryFreie Universitat BerlinThielallee 6314195BerlinGermany Institute of Biotechnology Technische Universit¨at BerlinStraße des 17.Juni 13510623BerlinGermany New/era/mabs GmbH August-Bebel-Str.8914482PotsdamGermany

Chinese hamster ovary(CHO)cells are crucial in biopharmaceutical production due to their scalability and capacity for human-like post-translational ***,toxic proteins and membrane proteins are often difficult-to-express in living ***,cell-free protein synthesis can be *** study explores innovative strategies for enhancing the production of challenging proteins through the modification of CHO cells by investigating both,cell-based and cell-free approaches.A major result in our study involves the integration of a mutant eIF2 translation initiation factor and T7 RNA polymerase into CHO cell lysates for cell-free protein *** resulted in elevated yields,while eliminating the necessity for exogenous additions during cell-free production,thereby substantially enhancing ***,we explore the potential of the Rosa26 genomic site for the integration of T7 RNA polymerase and cell-based tetracycline-controlled protein *** findings provide promising advancements in bioproduction technologies,offering flexibility to switch between cell-free and cell-based protein production as needed.

关键词： Inducible expression CHO cells Cell-free protein synthesis CRISPR T7 RNA polymerase eIF2 Rosa26

exploring the effects of dietary inulin in rainbow trout fed a high‑starch,100%plant‑based diet

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Journal of Animal Science and Biotechnology 2024年第2期15卷 713-732页

作者： Raphaël Defaix Jep Lokesh Laura Frohn Mickael Le Bechec Thierry Pigot Vincent Veron anne Surget Sandra Biasutti Frederic Terrier Sandrine Skiba-Cassy Jerome Roy Stephane Panserat Karine Ricaud Universite de Pau et Des Pays de L’Adour E2S UPPAINRAENUMEASaint‑Pee‑Sur‑NivelleFrance Universite de Pau et Des Pays de L’Adour E2S UPPACNRSIMT Mines AlesIPREMPauFrance Universite de Pau et Des Pays de L’Adour E2S UPPAIUT des Pays de l’AdourDepartement Genie BiologiqueMont de MarsanFrance.

Background High dietary carbohydrates can spare protein in rainbow trout(Oncorhynchus mykiss)but may affect growth and ***,a prebiotic,could have nutritional and metabolic effects,along with anti-inflammatory properties in teleosts,improving growth and *** tested this hypothesis in rainbow trout by feeding them a 100%plant-based diet,which is a viable alternative to fishmeal and fish oil in aquaculture *** a two-factor design,we examined the impact of inulin(2%)as well as the variation in the carbohydrates(CHO)/plant protein ratio on rainbow *** assessed the influence of these factors on zootechnical parameters,plasma metabolites,gut microbiota,production of short-chain fatty acids and lactic acid,as well as the expression of free-fatty acid receptor genes in the mid-intestine,intermediary liver metabolism,and immune markers in a 12-week feeding *** The use of 2%inulin did not significantly change the fish intestinal microbiota,but interestingly,the high CHO/protein ratio group showed a change in intestinal microbiota and in particular the beta diversity,with 21 bacterial genera affected,including Ralstonia,Bacillus,and 11 lactic-acid producing *** were higher levels of butyric,and valeric acid in groups fed with high CHO/protein diet but not with *** high CHO/protein group showed a decrease in the expression of pro-inflammatory cytokines(il1b,il8,and tnfa)in liver and a lower expression of the genes coding for tight-junction proteins in mid-intestine(tjp1a and tjp3).However,the 2%inulin did not modify the expression of plasma immune ***,inulin induced a negative effect on rainbow trout growth performance irrespective of the dietary *** With a 100%plant-based diet,inclusion of high levels of carbohydrates could be a promising way for fish nutrition in aquaculture through a protein sparing effect whereas the supplementation of 2%inulin does not appear to improve the use of CHO wh

关键词： Aquaculture Fish nutrition Gut microbiota Immune markers Intermediary metabolism Inulin Prebiotic Rainbow trout Short-chain fatty acids

Antecedents and sex/gender differences in youth suicidal behavior

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World Journal of Psychiatry 2014年第4期4卷 120-132页

作者： anne e rhodes Michael H Boyle Jeffrey A Bridge Mark Sinyor Paul S Links Lil Tonmyr Robin Skinner Jennifer M Bethell Corine Carlisle Sarah Goodday Travis Salway Hottes Amanda Newton Kathryn Bennett Purnima Sundar Amy H Cheung Peter Szatmari Institute for Clinical evaluative Sciences Toronto M4N 3M5 Ontario Canada Department of Psychiatry University of Toronto Toronto M5T 1R8 Ontario Canada Suicide Studies Unit St. Michael’s Hospital Toronto M5B 1W8 Ontario Canada Dalla Lana School of Public Health University of Toronto Toronto M5T 3M7 Ontario Canada Department of Psychiatry and Behavioural Neurosciences Mc Master University Hamilton L8N 3K7 Ontario Canada The Offord Centre for Child Studies Hamilton L8S 4K1 Ontario Canada The Research Institute at Nationwide Children’s Hospital Columbus OH 43205 United States The Ohio State University College of Medicine Columbus OH 43210 United States Sunnybrook Health Sciences Centre Toronto M4N 3M5 Ontario Canada Department of Psychiatry University of Western Ontario London N6A 5W9 Ontario Canada The Injury and Child Maltreatment Section Public Health Agency of Canada Ottawa K1A 0K9 Ontario Canada The Centre for Addiction and Mental Health Toronto M6J 1H4 Ontario Canada Department of Pediatrics University of Alberta Edmonton Alberta T6G 1C9 Canada The Department of Clinical epidemiology and Biostatistics Mc Master University Hamilton L8S 4K1 Ontario Canada The Ontario Centre of excellence for Child and Youth Mental Health Ottawa K1G 0Z1 Ontario Canada The Hospital for Sick Children Toronto M5G 1X8 Ontario Canada

Suicide is the second leading cause of death in youth globally; however, there is uncertainty about how best to intervene. Suicide rates are typically higher in males than females, while the converse is true for suicide attempts. We review this "gender paradox" in youth, and in particular, the age-dependency of these sex/gender differences and the developmental mechanisms that may explain them. epidemiologic, genetic,neurodevelopmental and psychopathological research have identified suicidal behaviour risks arising from genetic vulnerabilities and sex/gender differences in early adverse environments, neurodevelopment, mental disorder and their complex interconnections. Further, evolving sex-/gender-defined social expectations and norms have been thought to influence suicide risk. In particular, how youth perceive and cope with threats and losses(including conforming to others' or one's own expectations of sex/gender identity) and adapt to pain(through substance use and helpseeking behaviours). Taken together, considering brain plasticity over the lifespan, these proposed antecedents to youth suicide highlight the importance of interventions that alter early environment(s)(e.g., childhood maltreatment) and/or one's ability to adapt to them. Further, such interventions may have more enduring protective effects, for the individual and for future generations, if implemented in youth.

关键词： Suicide Attempted suicide Sex Gender Child Adolescent Review

Associations of accelerometry-measured and self-reported physical activity and sedentary behavior with skeletal muscle energetics:The Study of Muscle,Mobility and Aging(SOMMA)

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Journal of Sport and Health Science 2024年第5期13卷 621-630页

作者： Yujia(Susanna)Qiao Terri L.Blackwell Peggy M.Cawthon Paul M.Coen Steven R.Cummings Giovanna Distefano Samaneh Farsijani Daniel e.Forman Bret H.Goodpaster Stephen B.Kritchevsky Theresa Mau Frederico G.S.Toledo anne B.Newman Nancy W.Glynn Department of epidemiology School of Public HealthUniversity of PittsburghPittsburghPA 15261USA San Francisco Coordinating Center California Pacific Medical Center Research InstituteSan FranciscoCA 94143USA Department of epidemiology and Biostatistics University of CaliforniaSan FranciscoCA 94143USA Translational Research Institute AdventHealthOrlandoFL 32804USA Department of Medicine(Geriatrics and Cardiology) University of Pittsburghand GeriatricsResearchEducationand Clinical Center(GRECC)VA Pittsburgh Healthcare SystemPittsburghPA 15261USA Department of Internal Medicine Section on Gerontology and Geriatric MedicineWake Forest School of MedicineWinston-SalemNC 27109USA Division of endocrinology and Metabolism Department of MedicineUniversity of PittsburghPittsburghPA 15261USA

Background:Skeletal muscle energetics decline with age,and physical activity(PA)has been shown to offset these declines in older ***,many studies reporting these effects were based on self-reported PA or structured exercise ***,we examined the associations of accelerometry-measured and self-reported PA and sedentary behavior(SB)with skeletal muscle energetics and explored the extent to which PA and sedentary behavior would attenuate the associations of age with muscle ***:As part of the Study of Muscle,Mobility and Aging,enrolled older adults(n=879),810(age=76.4±5.0 years old,mean±SD;58%women)had maximal muscle oxidative capacity measured ex vivo via high-resolution re spirometry of permeabilized myofibers(maximal oxidative phosphorylation(maxOXPHOS))and in vivo by ^(31)phosphorus magnetic resonance spectroscopy(maximal adenosine triphosphate(ATP_(max))).Accelerometry-measured sedentary behavior,light activity,and moderate-to-vigorous PA(MVPA)were assessed using a wrist-worn ActiGraph GT9X over 7 ***-reported sedentary behavior,MVPA,and all PA were assessed with the Community Healthy Activities Model Program for Seniors(CHAMPS)*** regression models with progressive covariate adjustments evaluated the associations of sedentary behavior and PA with muscle energetics,as well as the attenuation of the age/muscle energetics association by MVP A and sedentary *** a sensitivity analysis,we also examined activPAL-measured daily step count and time spent in sedentary behavior and their associations with muscle ***:every 30 min/day more of ActiGraph-measured MVPA was associated with 0.65 pmol/(s×mg)higher maxOXPHOS and 0.012 mM/s higher ATP_(max)after adjusting for age,site/technician,and sex(ped with maxOXPHOS or ATP_(max).Meanwhile,every 30 min/day spent in ActiGraph-measured sedentary behavior was associated with 0.39 pmol/s×mg lower maxOXPHOS and0.

关键词： Aging exercise Mitochondria

evaluation of the Heat,entropy,and Rotational Changes Produced by Gravitational Segregation during Core Formation

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Journal of earth Science 2015年第1期26卷 124-133页

作者： anne M.Hofmeister Robert e.Criss Department of earth and Planetary Sciences Washington University

Core formation by gravitational segregation allegedly released sufficient interior heat to melt the earth. Analysis of the energetics, which compare gravitational potential energy （Ug） of a fictitious, homogeneous reference state to earth＇s current layered configuration, needs updating to correct errors and omissions, and to accommodate recent findings：（1） An erroneous positive sign was used for Ug while maintaining the reference value of 0 at infinity, which results in an incorrect sign fur AUg, which is crucial in determining whether a process is endothermic or exothermic. （2） The value of Ug for earth＇s initial state is uncertain. （3） Recent meteorite evidence indicates that core formation began before the earth was full-sized, which severely limits AUg. （4） Inhomogeneous accretion additionally reduced AUg. （5） The potentially large effect of differential rotation between the core and the mantle was not accounted for. （6） entropy changes associated with creating order were neglected. Accordingly, we revise values of Ug, evaluate uncertainties, and show that AUg was converted substantially to eonfigurational energy （TAS）. These considerations limit the large sources of primordial heat to impacts and radioactivity. Although these processes may play a role in core formation, their energies are independent of gravitational segregation, which produces order and rotational energy, not internal heat. Instead, gravitational segregation promotes planetary cooling mainly because it segregates lithophilic radioactive elements upward, increasing surface heat flux while shortening the distance over which radiogenic heat diffuses outwards.

关键词： core formation primordial heat configurational energy thermodynamics rotational energy.

Gradients in predation risk in a tropical river system

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Current Zoology 2018年第2期64卷 213-221页

作者： Amy e. DeACON Faith A. M. JONeS anne e. MAGURRAN Department of Life Sciences The University of the West Indies St Augustine Trinidad and Tobago School of Biology University of St Andrews Fife KY16 9TH Scotland UK

The importance of predation risk as a key driver of evolutionary change is exemplified by the Northern Range in Trinidad, where research on guppies living in multiple parallel streams has pro- vided invaluable insights into the process of evolution by natural selection. Although Trinidadian guppies are now a textbook example of evolution in action, studies have generally categorized predation as a dichotomous variable, representing high or low risk. Yet, ecologists appreciate that community structure and the attendant predation risk vary substantially over space and time. Here, we use data from a longitudinal study of fish assemblages at 16 different sites in the Northern Range to quantify temporal and spatial variation in predation risk. Specifically we ask： 1） Is there evidence for a gradient in predation risk？ 2） Does the ranking of sites （by risk） change with the defi- nition of the predator community （in terms of species composition and abundance currency）, and 3） Are site rankings consistent over time？ We find compelling evidence that sites lie along a contin- uum of risk. However, site rankings along this gradient depend on how predation is quantified in terms of the species considered to be predators and the abundance currency is used. Nonetheless, for a given categorization and currency, rankings are relatively consistent over time. Our study sug- gests that consideration of predation gradients will lead to a more nuanced understanding of the role of predation risk in behavioral and evolutionary ecology. It also emphasizes the need to justify and report the definition of predation risk being used.

关键词： abundance currency gradients Poecilia reticulata predation risk Trinidad Trinidadian guppy.

epidemiological, Clinical Progress Aspects of Congenital Heart Disease with Neonatal Revelation at the Mother-Child Hospital of Bingerville (HMe) Concerning 98 Cases from January 2021 to December 2022 (Côte d’Ivoire)

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Open Journal of Pediatrics 2024年第1期14卷 89-100页

作者： Adja evelyne Akaffou-Gbery Richard Azagoh-Kouadio anne-Lise Laetitia Mobio Jean-Jacques Yao Atteby Serenah Marie Janice N’vrah Anoh Kouassi Raoul Yao Department of Paediatrics and Neonatology H&#244pital Mère-Enfant de Bingerville (HME) Bingerville C&#244te d’Ivoire Department of Medical Pediatrics and Specialties CHU de Angré Abidjan C&#244te d’Ivoire

Introduction The distribution of congenital heart disease (CHD) in sub-Saharan Africa is highly imprecise and varies from one region to another due to the inequality of diagnostic facilities. The aim of this study was to determine the in-hospital prevalence of congenital heart disease in children at the Mother-Child hospital of Bingerville (HMe) by specifying the diagnostic, therapeutic and evolutionary aspects. Materials and methods We conducted a retrospective, descriptive, cross-sectional study at HMe of Bingerville from January 2021 to December 2022. All newborns with congenital heart disease confirmed by echocardiography were included in the study. Results Of 656 admissions to the neonatology department over the study period, congenital heart disease accounted for 14.9% (98/656) of cases. In our series, 76.7% were diagnosed before the 1st week of life, with a mean chronological age of 5.18 days and extremes of 0 and 46 days. There were as many male patients (50%) as female (50%), i.e. a sex ratio of 1. These newborns were premature in 60.2% of cases, with a mean and median gestational age of 34 weeks’ amenorrhea. Most were left-right shunts (90.8%). Persistent ductus arteriosus (PDA) (48.9%) predominated, followed by atrial septal defect (38.7%), ventricular septal defect (13.3%), common trunk artery (CTA) (3.1%) and open septal pulmonary atresia (OSPA) (1%) as the primary cyanogenic heart disease. Pulmonary arterial hypertension (PAH) (50%) was primary in 38.8% and secondary (61.2%). The mortality rate was 30.6%, and all CTA patients died (100%), with a significant statistical relationship (p = 0.027). Progression under treatment was marked by clinical stabilization (68/98) in 69.4% of cases. Conclusion: Congenital heart disease is relatively common at the Bingerville HMe. Access to echocardiography should be facilitated in neonatology departments for rapid diagnosis and optimal management of congenital heart disease in newborns.

关键词： Newborn Congenital Heart Disease Côte d’Ivoire

Can Modern Science Answer the Great Questions?

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Journal of earth Science 2022年第5期33卷 1330-1332页

作者： Robert e.Criss anne M.Hofmeister Dept.earth and Planetary Science Washington UniversitySt.LouisMO63130USA

0 INTRODUCTION Crucial issues facing us are whether current practices prohibit today’s scientists from answering great questions,and how to sufficiently educate the greater population so that scientific methods and f... 详细信息

关键词： Great finding facing

Identification of a novel population of human cord blood cells with hema-topoietic and chondrocytic potential

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Cell Research 2004年第4期14卷 268-282页

作者： Karen e JAY,anne ROULeAU,T Michael UNDeRHILL,Mickie BHATIA Stem Cell Biology and Regenerative Medicine The John P. Robarts Research Institute 100 Perth Drive London Ontario N6A 5K8 Canada Department of Microbiology and Immunology The University of Western Ontario London Ontario Canada Stem Cell Biology and Regenerative Medicine The John P. Robarts Research Institute 100 Perth Drive London Ontario N6A 5K8 Canada Department of Physiology The University of Western Ontario London Ontario Canada Stem Cell Biology and Regenerative Medicine The John P. Robarts Research Institute 100 Perth Drive London Ontario N6A 5K8 Canada Department of Microbiology and Immunology The University of Western Ontario London Ontario Canada. Department of Physiology The University of Western Ontario London Ontario Canada

With the exception of mature erythrocytes, cells within the human hematopoietic system are characterized by the cell surface expression of the pan-leukocyte receptor CD45. Here, we identify a novel subset among mononuclear cord blood cells depleted of lineage commitment markers (Lin) that are devoid of CD45 expression. Surprisingly, functional examination of Lin"CD45" cells also lacking cell surface CD34 revealed they were capable of multipotential hematopoietic progenitor capacity. Co-culture with mouse embryonic limb bud cells demonstrated that Lin-CD45-CD34- cells were capable of contributing to cartilage nodules and differentiating into human chondrocytes. BMP-4, a mesodermal factor known to promote chondrogenesis, significantly augmented Lin-CD45-CD34- differentiation into chondrocytes. Moreover, unlike CD34+ human hematopoietic stem cells, Lin-CD45-CD34- cells were unable to proliferate or survive in liquid cultures, whereas single Lin-CD45-CD34- cells were able to chimerize the inner cell mass (IC

关键词： hematopoietic chondrocyte CD45 embryonic.