Miracle-dz/UniFAR-FTUs huggingface dataset:
Identifying bottlenecks in transient and stable production of recombinant monoclonal‐antibody sequence variants in chinese hamster ovary cells

The increasing demand for antibody‐based therapeutics has emphasized the need for technologies to improve recombinant antibody titers from mammalian cell lines. Moreover, as antibody therapeutics address an increasing spectrum of indications, interest has increased in antibody engineering to improve affinity and biological activity. However, the cellular mechanisms that dictate expression and the relationships between antibody sequence and expression level remain poorly understood. Fundamental understanding of how mammalian cells handle high levels of transgene expression and of the relationship between sequence and expression are vital to the development of new antibodies and for increasing recombinant antibody titers. In this work, we analyzed a pair of mutants that vary by a single amino acid at Kabat position 49 (heavy‐chain framework), resulting in differential transient and stable titers with no apparent loss of antigen affinity. Through analysis of mRNA, gene copy number, intracellular antibody content, and secreted antibody, we found that while translational/post‐translational mechanisms are limiting in transient systems, it appears that the amount of available transgenic mRNA becomes the limiting event on stable integration of the recombinant genes. We also show that amino acid substitution at residue 49 results in production of a non‐secreted HC variant and postulate that stable antibody expression is maintained at a level which prevents toxic accumulation of this HC‐related protein. This study highlights the need for proper sequence engineering strategies when developing therapeutic antibodies and alludes to the early analysis of transient expression systems to identify the potential for aberrant stable expression behavior. © 2012 American Institute of Chemical Engineers Biotechnol. Prog. , 2012
Cross‐linked protein complex exhibiting asymmetric oxidation activities in the absence of added cofactor

A protein complex (PC) suspension exhibits asymmetric biooxidation activities in the absence of any added cofactor such as NAD(P)+ or FAD. It can be extracted from pea protein (PP)‐gel (PP encapsulated with Ca2+ alginate gel and aerated in air for several hours) using hot water by rotary shaking and powdered by the following three steps: (1) forming precipitates from the suspension using 30% (w/v) aqueous (NH4)2SO4, (2) crosslinking the precipitates with 0.25% (v/v) GA, and (3) preparing the cross‐linked powder by freeze‐drying. The cross‐linked PC (CLPC) performed asymmetric oxidation of the toward (R)‐isomers of rac‐1 and rac‐2 in 50 mM glycine–NaOH (pH 9.0) buffer/DMSO cosolvent [2.07% (v/v)] with high enantioselectivity; thus, the (S)‐isomers can be obtained in greater than 99% ee from the corresponding rac‐p‐substituted naphthyl methyl carbinol (rac‐1 and rac‐2). The CLPC activity was not only competitively inhibited by addition of either 1.0 mM ZnCl2 or a chelating agent such as 1.0 mM EDTA but also denatured by pretreatments: autoclaving at 121°C (20 min) or using 6.0 M guanidine–HCl containing 50 mM DTT. These results indicated that the PC catalytic process may utilize an electron transfer system incorporating a redox cation (e.g., Fe2+ ⇄ Fe3+ or Zn). Therefore, the newly introduced CLPC can asymmetrically oxidize the substrates without the addition of any cofactor resulting in a low‐cost organic method. Overall, our results show that the CLPC is an easily prepared, low‐cost reagent that can function under mild conditions and afford stereoselectivity, regioselectivity, and substrate specificity. © 2012 American Institute of Chemical Engineers Biotechnol. Prog. , 28: 953–961, 2012
The affective response to health-related information and its relationship to health anxiety: An ambulatory approach

Affective reactions to health-related information play a central role in health anxiety. Therefore, using ambulatory assessment, we analysed the time course of negative affect in a control group (CG, n = 60) which only rated their negative affect and an experimental group (EG, n = 97) which also rated the presence of somatic symptoms (e.g., back pain). By means of mixed regression models, we observed a decline of negative affect following the symptom self-ratings in the EG and a stable affect in the CG. The decline of negative affect was not moderated by the degree of health anxiety. Our findings might indicate that evaluating one's health status leads to a general reduction of negative affect in healthy individuals. The results of the study are in line with a bidirectional symptom perception model and underline the crucial role of affect regulation in the processing of health-related information.
Chemotherapy in metastatic renal cell carcinoma today? A systematic review

The prognosis of patients affected by metastatic renal cell carcinoma (mRCC) has improved markedly with targeted therapies. Unfortunately, 20–25% of the patients are refractory to treatment at the first response assessment and most patients will acquire drug resistance during the treatment. Moreover, current data on the clinical activity of targeted agents in poor risk or non-clear-cell mRCC patients are inconclusive because of the absence of prospective trials. Therefore, there are still several patients in need of new therapeutic approaches to improve clinical outcomes. Kidney cancer is historically considered resistant to chemotherapy on the basis that the results of phase II trials have not always been promising. We carried out a systematic review of both monochemotherapy and polychemotherapy alone or combined with immunotherapy or targeted agents in mRCC to define the state of the art and to evaluate further clinical research fields. All retrospectives, phase I/dose finding, phase II and phase III studies on chemotherapy in mRCC, published in the literature from January 2003 to November 2012, with at least 20 patients enrolled, were evaluated. Although the results of clinical trials have often been disappointing, in selected cases of mRCC, chemotherapy may have a promising antitumor activity, particularly when there are sarcomatoid differentiation features, or in highly progressive disease where the combination of doxorubicine plus gemcitabine or capecitabine has yielded interesting results. Chemotherapy may play a role in mRCC, whereas targeted agents and immunotherapy have not yielded durable and satisfactory results; further studies are needed.
Implications of Occupational Therapy Job Advertisement Trends for Occupational Therapy Education

Introduction: The aim of this study was to consider trends in occupational therapy workplace demands in Queensland between December 2009 and November 2011, as demonstrated by advertised job requirements. These trends are considered in the context of both undergraduate curriculum design and continuing professional development. Method: A mixed-methods approach was used. The content of occupational therapy job advertisements from four job databases over a 2-year period were examined using quantitative and qualitative analysis to identify trends, concepts, and themes. Results: Six lessons regarding occupational therapy education were drawn from the data. These included lessons relating to balancing graduate demand and supply, intake and exit timing, high-demand areas of practice, non-traditional occupational therapy roles, the roles of continuing professional development providers, and discrepancies between existing new graduate competencies and required workplace competencies. Conclusion: Occupational therapy educators have an obligation to produce graduates who meet the profession's graduate competencies, as well as those competencies required by workplaces, if those graduates are to be considered work-ready and fit for practice. Similarly, education at a continuing professional development level should reflect market trends and offer graduates and practitioners the opportunity to maintain employment currency.
Optimization of a two‐step process comprising lipase catalysis and thermal cyclization improves the efficiency of synthesis of six‐membered cyclic carbonate from trimethylolpropane and dimethylcarbonate

Six‐membered cyclic carbonates are potential monomers for phosgene and/or isocyanate free polycarbonates and polyurethanes via ring‐opening polymerization. A two‐step process for their synthesis comprising lipase‐catalyzed transesterification of a polyol, trimethylolpropane (TMP) with dimethylcarbonate (DMC) in a solvent‐free system followed by thermal cyclization was optimized to improve process efficiency and selectivity. Using full factorial designed experiments and partial least squares (PLS) modeling for the reaction catalyzed by Novozym®435 (N435; immobilized Candida antarctica lipase B), the optimum conditions for obtaining either high proportion of monocarbonated TMP and TMP‐cyclic‐carbonate (3 and 4), or dicarbonated TMP and monocarbonated TMP‐cyclic‐carbonate (5 and 6) were found. The PLS model predicted that the reactions using 15%–20% (w/w) N435 at DMC:TMP molar ratio of 10–30 can reach about 65% total yield of 3 and 4 within 10 h, and 65%–70% total yield of 5 and 6 within 32–37 h, respectively. High consistency between the predicted results and empirical data was shown with 66.1% yield of 3 and 4 at 7 h and 67.4% yield of 5 and 6 at 35 h, using 18% (w/w) biocatalyst and DMC:TMP molar ratio of 20. Thermal cyclization of the product from 7 h reaction, at 110°C in the presence of acetonitrile increased the overall yield of cyclic carbonate 4 from about 2% to more than 75% within 24 h. N435 was reused for five consecutive batches, 10 h each, to give 3+4 with a yield of about 65% in each run. © 2012 American Institute of Chemical Engineers Biotechnol. Prog. , 2013.
Trials of biomarker-guided antimicrobial therapy in sepsis: improvements in trial design require better control groups.

e172 www.ccmjournal. org February 2014 • Volume 42 • Number 2 Dr. Palomar lectured for Pfizer and 3M. Dr. Lerma received support for travel from Ministerio Sanidad; consulted for Pfizer, Astellas, Novartis, and Gilead; received grant support from MSD; lectured for Pfizer, MSD, and Astellas; and received support for the development of educational presentations from Astellas; and received support for travel from Pfizer. His institution received grant support from Pfizer, Astellas, and Novartis. Dr. Agra disclosed that she does not have any potential conflicts of interest.
Mastering moral misery: Emotional and coping responses to intragroup morality (vs. competence) evaluations

In social groups, individuals are often confronted with evaluations of their behaviour by other group members and are motivated to adapt their own behaviour accordingly. In two studies we examine emotional responses towards, and perceived coping abilities with, morality vs. competence evaluations individuals receive from other in-group members. In Study 1, we show that evaluations of one's immoral behaviour primarily induce guilt, whereas evaluations of incompetent behaviour raise anger. In Study 2, we elaborate on the psychological process associated with these emotional responses, and demonstrate that evaluations of immorality, compared to incompetence, diminish group members’ perceived coping abilities, which in turn intensifies feelings of guilt. However, when anticipating an opportunity to restore one's self-image as a moral group member, perceived coping abilities are increased and the experience of guilt is alleviated. Together these studies demonstrate how group members can overcome their moral misery when restoring their self-image.
CD34 mediates intestinal inflammation in Salmonella‐infected mice

CD34 is a highly glycosylated sialomucin expressed on a variety of cells, ranging from vascular endothelial cells to haematopoietic stem cells. Depending on its glycosylation state, CD34 has been shown to promote or inhibit cell adhesion and migration; however, a functional role for CD34 in the gut has not been determined. Using a model of Salmonella‐induced gastroenteritis, we investigated the role of CD34 in the context of infection. Upon oral infection, the number of CD34+ cells detected in the submucosa, vascular endothelium and lamina propria significantly increased in S. Typhimurium‐infected C57Bl/6 mice. The pathology of S. Typhimurium‐infected C57Bl/6 mice was characterized by recruitment of neutrophils to the site of inflammation, submucosal oedema and crypt destruction. In contrast, Cd34−/− mice showed a delayed pathology, a defect in inflammatory cell migration into the intestinal tissue and enhanced survival. Importantly, this was not due to a lack of chemotactic signals in Cd34−/− mice as these mice had either similar or significantly higher levels of pro‐inflammatory cytokines and chemokines post infection when compared with infected C57/Bl6 control mice. In summary, we demonstrate a novel role for CD34 in enhancing migration of inflammatory cells and thereby exacerbating host‐mediated immunopathology in the intestine of S. Typhimurium‐infected mice.
Enzymatic digestion of alkaline‐sulfite pretreated sugar cane bagasse and its correlation with the chemical and structural changes occurring during the pretreatment step

Sugar cane bagasse is recalcitrant to enzymatic digestion, which hinders the efficient conversion of its polysaccharides into fermentable sugars. Alkaline‐sulfite pretreatment was used to overcome the sugar cane bagasse recalcitrance. Chemical and structural changes that occurred during the pretreatment were correlated with the efficiency of the enzymatic digestion of the polysaccharides. The first 30 min of pretreatment, which removed approximately half of the initial lignin and 30% of hemicellulose seemed responsible for a significant enhancement of the cellulose conversion level, which reached 64%. After the first 30 min of pretreatment, delignification increased slightly, and hemicellulose removal was not enhanced; however, acid groups continued to be introduced into the residual lignin. Water retention values were 145% to the untreated bagasse and 210% to the bagasse pretreated for 120 min and fiber widths increased from 10.4 to 30 μm, respectively. These changes were responsible for an additional increase in the efficiency of enzymatic hydrolysis of the cellulose, which reached 92% with the 120 min pretreated sample. © 2013 American Institute of Chemical Engineers Biotechnol. Prog. , 29:890–895, 2013
Entry of Bacillus anthracis spores into epithelial cells is mediated by the spore surface protein BclA, integrin α2β1 and complement component C1q

Inhalational anthrax is initiated by pulmonary exposure to Bacillus anthracis spores. Spore entry into lung epithelial cells is observed both in vitro and in vivo and evidence suggests it is important for bacterial dissemination and virulence. However the specific host receptor and spore factor that mediate the entry process were unknown. Here, we report that integrin α2β1 is a major receptor for spore entry. This is supported by results from blocking antibodies, siRNA knock‐down, colocalization, and comparison of spore entry into cells that do or do not express α2. BclA, a major spore surface protein, is found to be essential for entry and α2β1‐mediated entry is dependent on BclA. However, BclA does not appear to bind directly to α2. Furthermore, spore entry into α2‐expressing cells is dramatically reduced in the absence of serum, suggesting that additional factors are involved. Finally, complement component C1q, also an α2β1 ligand, appears to act as a bridging molecule or a cofactor for BclA/α2β1‐mediated spore entry and BclA binds to C1q in a dose‐dependent and saturable manner. These findings suggest a novel mechanism for pathogen entry into host cells as well as a new function for C1q–integrin interactions. The implications of these findings are discussed.
Starvation and rapamycin differentially regulate host cell lysosome exocytosis and invasion by Trypanosoma cruzi metacyclic forms

The molecular mechanisms of host cell invasion by T. cruzi metacyclic trypomastigotes (MT), the developmental forms that initiate infection in the mammalian host, are only partially understood. Here we aimed at further identifying the target cell components involved in signalling cascades leading to MT internalization, and demonstrate for the first time the participation of mammalian target of rapamycin (mTOR). Treatment of human epithelial HeLa cells with mTOR inhibitor rapamycin reduced lysosomal exocytosis and MT invasion. Downregulation of phosphatidylinositol 3‐kinase and protein kinase C also impaired exocytosis and MT internalization. The recombinant protein based on gp82, the MT surface molecule that mediates cell adhesion/invasion, induced exocytosis in HeLa cells. Such an effect has not previously been attributed to any T. cruzi surface molecule. Rapamycin treatment diminished gp82 binding as well. Cell invasion assays under conditions that promoted lysosome exocytosis, such as 1 h incubation in starvation medium PBS++, increased MT invasion, whereas pre‐starvation of cells for 1–2 h had an opposite effect. In contrast to MT, invasion of tissue culture trypomastigotes (TCT) increased upon host cell pre‐starvation or treatment with rapamycin, a novel finding that discloses quite distinctive features of the two infective forms in a key process for infection.
Altered developmental expression of polymorphic membrane proteins in penicillin‐stressed Chlamydia trachomatis

Late Chlamydia trachomatis inclusions express each member of the surface‐exposed polymorphic membrane protein family (Pmp subtypes A through I) with a reproducible distribution of fully‐on, fully‐off and intermediate phenotypes. This observation is consistent with observed variable Pmp antibody profiles in C. trachomatis‐infected patients and has led to the hypothesis that the pmp gene family forms the basis of a phase variation‐like mechanism of antigenic variation. Here we investigate and compare the developmental expression of each of the nine pmp genes under conditions of optimal in vitro growth with that under conditions that promote prolonged survival of chlamydiae when exposed to penicillin‐induced stress. We demonstrate that the pmp gene family includes distinct transcriptional units that are differentially expressed along development and differentially responsive to stress. In particular, our results indicate that expression of pmpA, pmpD and pmpI is uniquely unaffected by stress, suggesting that the PmpA, PmpD and PmpI proteins play a critical role in the pathogenesis of C. trachomatis.
Effects of Ethnicity on Deceased Organ Donation in a Minority-Majority State*

Objective:To define how ethnicity affects donation rates in New Mexico when compared with the United States. We hypothesized that deceased donation rates in New Mexico would reflect the ethnic rates of the population. Design:We performed a retrospective review of the Organ Procurement Database for New Mexico from 2009 to June 2012. Methods:Rates for donors and transplant candidates were calculated relative to 2010 census population estimates by ethnicity for non-Hispanic Whites, Hispanics, and American Indians. Poisson regression analyses were used to test whether United States and New Mexico rates differed. Rates were scaled to 100,000 patient-years for reporting. Setting:State of New Mexico population compared to United States population. Subjects:Reported deaths to New Mexico Donor Services and United Network for Organ Sharing from 2009 to 2012. Interventions:None. Measurements and Main Results:Non-Hispanic White age-adjusted donor rates per 100,000 patient-years were 2.58 in New Mexico versus 2.60 in the United States, Hispanic donor rates were 1.98 in New Mexico versus 2.03 nationwide, and American Indian donor rates in New Mexico were 0.26 versus 1.23 nationwide (rate ratio = 0.21; 95% CI, 0.05–0.86). American Indians have significantly lower donor rates in New Mexico compared to non-Hispanic Whites (rate ratio = 0.11) and Hispanics (rate ratio = 0.13) and nationally (non-Hispanic Whites: rate ratio = 0.32 and Hispanics: rate ratio = 0.43). Hispanics and non-Hispanic Whites had similar donor rates regardless of geographic strata (Hispanics vs non-Hispanic Whites, New Mexico: 0.83; United States: 0.75). In New Mexico, Hispanic patients were 1.43 times more likely to be listed as transplant candidates than non-Hispanic Whites and American Indians were 3.32 times more likely to be listed than non-Hispanic Whites. In the United States, Hispanic patients were 1.90 times more likely to be listed as transplant candidates than non-Hispanic Whites and American Indians were 1.55 times more likely to be listed than non-Hispanic Whites. Conclusions:Donor and transplant candidate rates did not show strong differences by geographic strata. These findings suggest that further work is needed to elucidate the causes for ethnic differences in rates of consent and donation, particularly in the American Indian population.
Cardiovascular Magnetic Resonance in Myocarditis: A JACC White Paper

2009;53;1475-1487 J. Am. Coll. Cardiol. Consensus Group on Cardiovascular Magnetic Resonance in Myocarditis Filipchuk, Andreas Kumar, Matthias Pauschinger, Peter Liu, for the International Gutberlet, Sanjay Prasad, Anthony Aletras, Jean-Pierre Laissy, Ian Paterson, Neil G. Pauline Alakija, Leslie T. Cooper, James A. White, Hassan Abdel-Aty, Matthias Matthias G. Friedrich, Udo Sechtem, Jeanette Schulz-Menger, Godtfred Holmvang, Cardiovascular Magnetic Resonance in Myocarditis: A JACC White Paper This information is current as of April 21, 2009 http://content.onlinejacc. org/cgi/content/full/53/17/1475 located on the World Wide Web at: The online version of this article, along with updated information and services, is
Attentional capture by irrelevant emotional distractor faces is contingent on implicit attentional settings

ABSTRACT Although expressions of facial emotion hold a special status in attention relative to other complex objects, whether they summon our attention automatically and against our intentions remains a debated issue. Studies supporting the strong view that attentional capture by facial expressions of emotion is entirely automatic reported that a unique (singleton) emotional face distractor interfered with search for a target that was also unique on a different dimension. Participants could therefore search for the odd-one out face to locate the target and attentional capture by irrelevant emotional faces might be contingent on the adoption of an implicit set for singletons. Here, confirming this hypothesis, an irrelevant emotional face captured attention when the target was the unique face with a discrepant orientation, both when this orientation was unpredictable and when it remained constant. By contrast, no such capture was observed when the target could not be found by monitoring displays for a discrepant face and participants had to search for a face with a specific orientation. Our findings show that attentional capture by emotional faces is not purely stimulus driven and thereby resolve the apparent inconsistency that prevails in the literature on the automaticity of attentional capture by emotional faces.