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Good teaching requires pedagogical dynamism: a willingness to vary one's teaching approach relative to the context (and cohort) at hand, and to any new challenges that may arise from that context. This requires that teachers obtain a broad knowledge of teaching strategies and tactics. Given the demands of contemporary higher education, finding the time to obtain this knowledge can pose a challenge to full‐time academics. This is exacerbated in the case of part‐time and practitioner‐teachers who often work in varying degrees of isolation from the communities of learning to which they contribute. An analysis of the teaching practices of eight high‐performing practitioner‐teachers, alongside data obtained from 74 students, within a master's‐level architectural course at a large Australian university, has been used to delineate five strategies for optimising student learning within the architecture studio. These include: setting up a challenge that clearly articulates the role of the student within it; early and frequent assessment tasks to assist students in rapid skill development and reflective practice; communicating clear expectations around performance; facilitating a strong peer culture; and enhancing motivation via high expectations. This article details the tactics by which these strategies can be achieved. This article delivers directly applicable advice for architectural educators, but it also provides practical insight into the specifics of architectural teaching that may assist educators working in other disciplines. [ABSTRACT FROM AUTHOR]

Microbiota derived metabolites act as chemical messengers that elicit a profound impact on host physiology. Vitamin D receptor (VDR) is a key genetic factor for shaping the host microbiome. However, it remains unclear how microbial metabolites are altered in the absence of VDR. We investigated metabolites from mice with tissue-specific deletion of VDR in intestinal epithelial cells or myeloid cells. Conditional VDR deletion severely changed metabolites specifically produced from carbohydrate, protein, lipid, and bile acid metabolism. Eighty-four out of 765 biochemicals were significantly altered due to the Vdr status, and 530 significant changes were due to the high-fat diet intervention. The impact of diet was more prominent due to loss of VDR as indicated by the differences in metabolites generated from energy expenditure, tri-carboxylic acid cycle, tocopherol, polyamine metabolism, and bile acids. The effect of HFD was more pronounced in female mice after VDR deletion. Interestingly, the expression levels of farnesoid X receptor in liver and intestine were significantly increased after intestinal epithelial VDR deletion and were further increased by the high-fat diet. Our study highlights the gender differences, tissue specificity, and potential gut-liver-microbiome axis mediated by VDR that might trigger downstream metabolic disorders. [ABSTRACT FROM AUTHOR]

Universal school enrolment of children aged 6–14 years has been a commitment enshrined in the Indian constitution since 1950. To attain this goal, the central government has set up a range of programs, such as incentive programs, teacher grants and infrastructure grants, while state governments share responsibility in implementing those educational policy initiatives. This paper reviews the progress of India’s primary schools over the period 2005–2011 as public educational undertaking shifted more from the centre to the states. Using two waves of the Indian Human Development Survey conducted in 2005 and 2011, we track the progress of primary schools in terms of infrastructure upgrade, teachers’ quality, the costs of education, enrolment and learning outcomes. We find that while progress has been made in terms of infrastructure provision and enrolment, other aspects have either seen no progress or have worsened over time, and numerical and reading skills of children 8–11 years old worsening over time. Better performing states are generally in the South and in the extreme North. However, in terms of learning outcomes, the abysmal results are nationwide. [ABSTRACT FROM AUTHOR]

The nursing role is rapidly evolving as nurses are tasked with an even wider range of healthcare responsibilities. With an increase in complexity and demand for better healthcare system, the roles of nurses have become more challenging. A poor mental health and quality of sleep of nurses can have an impact on their day to day functioning which in turn can pose a threat, especially on the patients of critical wards as compared to other wards, because they require complex assessment and high risk treatments. Moreover, nurses are often scheduled to do rotating shifts (i.e., day, evening, night) and also have to work overtime to provide quality care to their patients around the clock. Abrupt deviations from normal timing of work often disrupt one's internal biological clock. The present study aims to trace and compare the dimensions of positive mental health status and quality of sleep among nurses of critical and general ward of private hospitals with an age of 21 to 31yrs. To collect data, quantitative and survey research was conducted. The result showed no significant difference between the nurses of two wards with regards to their mental health status. But on the other hand, the critical ward nurses showed poor quality of sleep which is more related to 'sleep duration' and 'sleep latency'. The study can be important for the administrative staffs of private hospitals, so that they can recommend and make arrangements for nurses and their duties accordingly. [ABSTRACT FROM AUTHOR]

Infection with the protozoan parasite Cryptosporidium parvum (CP) causes cryptosporidiosis, a widespread diarrhoeal disease. Impaired intestinal epithelial barrier function and increased permeability are most commonly associated with diarrhoeal diseases caused by enteric infections. However, studies on barrier disruption and underlying mechanisms in cryptosporidiosis are extremely limited. Epithelial tight junctions (TJs) and adherens junctions (AJs) are important in maintaining barrier integrity. Therefore, we examined the effects of CP infection on paracellular permeability and on the expression of the major TJ and AJ proteins utilising in vitro, ex vivo, and in vivo models. CP infection (0.5 × 106 oocysts/well in Transwell inserts, 24 hr) increased paracellular permeability (FITC-dextran flux) in Caco-2 cell monolayers and substantially decreased the protein levels of occludin, claudin 4, and E-cadherin. Claudin 3, zonula occludens-1 (ZO1) and α-catenin were also significantly decreased, whereas claudins 1 and 2 and β-catenin were not altered. Substantial downregulation of occludin, claudin 4, and E-cadherin was also observed in response to CP infection ex vivo in mouse enteroid-derived monolayers and in vivo in the ileal and jejunal mocosa of C57BL/6 mice. The mRNA levels of these proteins were also significantly decreased in CP-infected mouse ileum and jejunum but were unaltered in Caco-2 cells. Further, bafilomycin-A, an inhibitor of lysosomal proton pump, partially abrogated CP effects on occludin expression in Caco-2 cells, suggesting a potential role of posttranslational mechanisms, such as induction of protein degradation pathways, in mediating the effects of the parasite. Our studies suggest that disruption of barrier function via downregulation of specific key components of TJ and AJ could be a major mechanism underlying CP infection-induced diarrhoea. [ABSTRACT FROM AUTHOR]

In India, millions of female foetuses have been aborted since the 1980s alongside an abnormally high infant girl mortality rate; this has generated a vast literature exploring the root causes of son preference. The literature is sparse, however, on how the decisions to abort or neglect girls are made. This paper examines mothers’ and fathers’ respective roles behind those decisions. Using the third National Family and Health Survey (NFHS-3) data, we show that sex-selective abortions are most commonly used if both spouses or if only the fathers prefer sons, while sex-selective neglect is used if only the mothers prefer sons. [ABSTRACT FROM PUBLISHER]

In the present study, we have examined the impact of Age, Gender and Socio-economic status on Morality and Valued living of300 individuals belonging to four zones (North, South, East, & West) of Kolkata City. The results of the study revealed that (a) age of the individuals seems to have a significant impact on their morality and valued living, (b) gender of the individuals seems to have a significant impact on their morality and valued living and (c) socio-economic status of the individuals seems to have a significant impact on their morality but not on their valued living. The present study appears to be quite relevant in the present scenario of our society. The implication of the study lies in taking proper care for the inculcation of moral values among the youth as they are the future of our society. [ABSTRACT FROM AUTHOR]

SLC26A3 [downregulated in adenoma (DRA)] plays a key role in mammalian intestinal NaCl absorption, in that it mediates apical membrane Cl-/HCO3- exchange. DRA function and expression are significantly decreased in diarrhea associated with inflammatory bowel disease. DRA is also considered to be a marker of cellular differentiation and is predominantly expressed in differentiated epithelial cells. Caudal-type homeobox protein-2 (CDX2) is known to regulate genes involved in intestinal epithelial differentiation and proliferation. Reduced expression of both DRA and CDX2 in intestinal inflammation prompted us to study whether the DRA gene is directly regulated by CDX2. Our initial studies utilizing CDX2 knockout (CDX2fV/fV;Cre+) mice showed a marked reduction in DRA mRNA and protein levels in proximal and distal colon. In silico analysis of the DRA promoter showed two consensus sites for CDX2 binding. Therefore, we utilized Caco-2 cells as an in vitro model to examine if DRA is a direct target of CDX2 regulation. siRNAmediated silencing of CDX2 in Caco-2 cells resulted in a marked (~50%) decrease in DRA mRNA and protein levels, whereas ectopic overexpression of CDX2 upregulated DRA expression and also stimulated DRA promoter activity, suggesting transcriptional regulation. Electrophoretic mobility shift and chromatin immunoprecipitation assays demonstrated direct binding of CDX2 to one of the two putative CDX2 binding sites in the DRA promoter (+645/+663). In summary, our studies, for the first time, demonstrate transcriptional regulation of DRA expression by CDX2, implying that reduced expression of DRA in inflammatory bowel disease-associated diarrhea may, in part, be due to downregulation of CDX2 in the inflamed mucosa. [ABSTRACT FROM AUTHOR]

Aims Lipid phosphate phosphatase-3 (LPP3) is expressed at high levels in endothelial cells (ECs). Although LPP3 is known to hydrolyse the phosphate group fromlysolipids such as spingosine-1-phosphate and its structural homologues, the function of Lpp3 in ECs is not completely understood. In this study, we investigated how tyrosine-protein kinase receptor (TEK or Tie2) promoter-dependent deletion of Lpp3 alters EC activities. Methods and results Lpp3fl/fl micewere crossedwith the tg.Tie2Cre transgenic line. Vasculogenesis occurred normally in embryos with Tie2Cre mediated deletion of Lpp3 (called Lpp3ECKO), but embryonic lethality occurred in two waves, the first wave between E8.5 and E10.5, while the second between E11.5 and E13.5. Lethality in Lpp3ECKO embryos after E11.5 was accompanied by vascular leakage and haemorrhage, which likely resulted in insufficient cardiovascular development. Analyses of haematoxylin- and eosin-stained heart sections from E11.5 Lpp3ECKO embryos showed insufficient heart growth associated with decreased trabeculation, reduced growth of the compact wall, and absence of cardiac cushions. Staining followed by microscopic analyses of Lpp3ECKO embryos revealed the presence of apoptotic ECs. Furthermore, Lpp3-deficient ECs showed decreased gene expression and protein levels of Cyclin-D1, VE-cadherin, Fibronectin, Klf2, and Klf4. To determine the underlying mechanisms of vascular leakage and barrier disruption, we performed knockdown and rescue experiments in cultured ECs. LPP3 knockdown decreased transendothelial electrical resistance and increased permeability. Re-expression of β-catenin cDNA in LPP3-knockdown ECs partially restored the effect of the LPP3 loss, whereas re-expression of p120ctn cDNA did not. Conclusion These findings demonstrate the essential roles of LPP3 in the maturation of EC barrier integrity and normal cardiovascular development. [ABSTRACT FROM AUTHOR]

This study was designed to investigate the relation between Materialistic Value, Spiritual Oneness Belief and Physical Oneness Belief with Happiness. Positive psychology define happiness as nothing else than an emotion, a long-term sense of emotional well-being and contentment - a broad "feeling" that one is happy. The sample consisted of 100 (50 males and 50 females) young professionals (public and private sectors), selected randomly from various organizations of Kolkata and Howrah districts. 3 adapted scales namely- Materialistic Value Scale by Richin (2004), Oneness Belief Scale by Garfield, Drwecki, Moore, Kortenkamp and Gracz (2012) and Oxford Happiness Questionnaire by Michael Argyle and Peter Hills (1998) were administered to the selected respondents. The results revealed that Materialistic Value was negatively and significantly correlated with Happiness and Physical Oneness Belief was positively and significantly correlated with Happiness. Spiritual Oneness Belief showed a mild positive correlation with Happiness. Linear regression revealed Materialistic Value to be a negative predictor of Happiness and it could predict only 11.5% of the variance. The study has applications in the field of positive psychology. [ABSTRACT FROM AUTHOR]

Background: The acquisition of proliferative and invasive phenotypes is considered a hallmark of neoplastic transformation; however, the underlying mechanisms are less well known. Lipid phosphate phosphatase-3 (LPP3) not only catalyzes the dephosphorylation of the bioactive lipid sphingosine-1-phosphate (S1P) to generate sphingosine but also may regulate embryonic development and angiogenesis via the Wnt pathway. The goal of this study was to determine the role of LPP3 in tumor cells. Results: We observed increased expression of LPP3 in glioblastoma primary tumors and in U87 and U118 glioblastoma cell lines. We demonstrate that LPP3-knockdown inhibited both U87 and U118 glioblastoma cell proliferation in culture and tumor growth in xenograft assays. Biochemical experiments provided evidence that LPP3-knockdown reduced β-catenin, CYCLIN-D1, and CD133 expression, with a concomitant increase in phosphorylated β-catenin. In a converse experiment, the forced expression of LPP3 in human colon tumor (SW480) cells potentiated tumor growth via increased β-catenin stability and CYCLIN-D1 synthesis. In contrast, elevated expression of LPP3 had no tumorigenic effects on primary cells. Conclusions: These results demonstrate for the first time an unexpected role of LPP3 in regulating glioblastoma progression by amplifying β-catenin and CYCLIN-D1 activities. [ABSTRACT FROM AUTHOR]

As governments around the globe navigate the effects of COVID‐19 crisis, the urban poor endure the unevenly distributed socio‐economic impacts of the pandemic. This burden is more pronounced in Global South megacities, where millions of people engage in precarious informal employment. We examine how the urban poor in Delhi (India), Dhaka (Bangladesh), and Manila (Philippines) have been disproportionately affected by the crisis. A cross‐case analysis was undertaken to determine how the realities and relations of one context can enrich our understanding of the other. We argue that the current COVID‐19 pandemic has exposed the unequal urban citizenship in these three metropolises. Drawing on research reports, news articles, and interviews with urban poor leaders, we explain how limited government assistance has forced some to retreat to their rural hometowns or rely on self‐help and mutual aid practices to survive. We consider the patterns both in governments' treatment of impoverished citizens and in the unjust effects of lockdown enforcement on marginalised populations. We also discuss the roles that women and non‐government organisations (NGOs) have played in facilitating solidarity‐based practices to help urban poor communities cope with COVID‐19 vulnerabilities. [ABSTRACT FROM AUTHOR]

To claim welfare entitlements, the residents challenge state enumeration procedures and carry out a multitude of documentary and inscriptive struggles in the city. Such "numerical strength" is also evident in Old Fadama, Accra, Ghana, where resident population numbers became highly contentious between residents and state officials, with community-led enumeration counting 2.5 times official figures (Farouk & Owusu, [8]). If the state agencies defer in issuing proper documents, the poor interrogate the politics of deferral and the dilatory tactics of the state by filing applications under the ambit of the Right to Information policy. The diverse research sites offer the opportunity to explore some of the similarities and differences among the various tactics that residents adopt to navigate their precarious living conditions and engage with state authorities. [Extracted from the article]

TGF-β1 is an important multifunctional cytokine with numerous protective effects on intestinal mucosa. The influence of TGF-β1 on serotonin transporter (SERT) activity, the critical mechanism regulating the extracellular availability of serotonin (5-HT), is not known. Current studies were designed to examine acute effects of TGF-β1 on SERT. Model human intestinal Caco-2 cells grown as monolayer’s or as cysts in 3D culture and ex vivo mouse model were utilized. Treatment of Caco-2 cells with TGF-β1 (10 ng/ml, 60 min) stimulated SERT activity (~2 fold, P<0.005). This stimulation of SERT function was dependent upon activation of TGF-β1 receptor (TGFRI) as SB-431542, a specific TGF-βRI inhibitor blocked the SERT stimulation. SERT activation in response to TGF-β1 was attenuated by inhibition of PI3K and occurred via enhanced recruitment of SERT-GFP to apical surface in a PI3K dependent manner. The exocytosis inhibitor brefeldin A (2.5 μM) attenuated the TGF-β1-mediated increase in SERT function. TGF-β1 increased the association of SERT with the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) syntaxin 3 (STX3) and promoted exocytosis of SERT. Caco-2 cells grown as cysts in 3D culture recapitulated the effects of TGF-β1 showing increased luminal staining of SERT. Ussing chamber studies revealed increase in 3H-5-HT uptake in mouse ileum treated ex vivo with TGF-β1 (10 ng/ml, 1h). These data demonstrate a novel mechanism rapidly regulating intestinal SERT via PI3K and STX3. Since decreased SERT is implicated in various gastro-intestinal disorders e.g IBD, IBS and diarrhea, understanding mechanisms stimulating SERT function by TGF-β1 offers a novel therapeutic strategy to treat GI disorders. [ABSTRACT FROM AUTHOR]

Abstract: In this review, we discuss the role of focal adhesion kinase (FAK), an intracellular tyrosine kinase, in endothelial cells in relation to neovascularization. Genetic and in vitro studies have identified critical factors, receptor systems, and their intracellular signaling components that regulate the neovasculogenic phenotypes of endothelial cells. Among these factors, FAK appears to regulate several aspects of endothelial cellular behavior, including migration, survival, cytoskeletal organization, as well as cell proliferation. Upon adhesion of endothelial cells to extracellular matrix (ECM) ligands, integrins cluster on the plane of plasma-membrane, while cytoplasmic domains of integrins interact with cytoskeletal proteins and signaling molecules including FAK. However, FAK not only serves as a critical component of integrin signaling, but is also a downstream element of the VEGF/VEGF–receptor and other ligand–receptor systems that regulate neovascularization. A complete understanding of FAK-mediated neovascularization, therefore, should address the molecular and cellular mechanisms that regulate the biology of FAK. Continued research on FAK may, therefore, yield novel therapies to improve treatment modalities for the pathological neovascularization associated with diseases. [Copyright &y& Elsevier]

Everybody knows that the phrase 'informal settlement' is a widely used euphemism for 'slum', avoiding a word that can be seen to cast a stigma on neighbourhoods of the urban poor. Yet we have also long known that 'informal settlement' is a verb – identifying self-organised modes of production through which the urban poor produce affordable housing and urban infrastructure. Here we argue four key points. First, the distinction between slums and informal settlement is evident in UN-Habitat data that suggests that informal settlement has rapidly expanded while slums have not. Second, informality is a paradoxical condition that can be identified with the illegality, insecurity and inferiority of the slum, but also with the incremental self-organisation of urban commoning and the right to the city. Third, while the language of the 'slum' will inevitably persist, the UN-Habitat definition is both too broad and too narrow – conflating material conditions with overcrowding and tenure insecurity while excluding overdevelopment. Finally, if we want to understand 'informal settlement' as a dominant mode of urban production then we need to stop using it interchangeably with the term 'slum' and engage with how it works as the design and planning of housing and urban infrastructure. [ABSTRACT FROM AUTHOR]

Informal settlement is the most pervasive mode of urbanization throughout cities of the global South, yet little is known of how informal urban design works as a production of urban space. This paper maps the emergence and transformation of street/laneway networks and buildings in a broad range of settlements. While such data cannot reveal the social, economic and political complexity of individual cases, it demonstrates the range of informal morphologies and self-organizational practices that produce them. Building types, plots, blocks, streets and lanes are analyzed to extract the informal rules or logics that are embodied within such morphologies. This is a nascent study that begins to reveal the ways in which informal settlement works to produce habitable land, affordable housing and public space. The prospect is a better knowledge base for understanding how such practices may or may not produce 'slums', how upgrading practices may be improved and how informal production may be anticipated and harnessed. [ABSTRACT FROM AUTHOR]

Natural products represent a rich source of biologically active compounds and an example of molecular diversity with recognized potential in drug discovery. Nowadays, medicinal mushrooms used in folk medicine are being increasingly studied for their successful application in pharmaceutical industries. In this context, Pleurotus species are one such potential mushroom with worldwide acceptance as a delicious food and famed for remarkable medicinal properties. However, till date no reports regarding the bioactivity of the most compatible herbal formulations of this mushroom (infusion and decoction) have been documented. Therefore, the present study has been undertaken to investigate the phytochemical composition and antioxidant properties of three cultivated species of this group namely, P. eous, P. florida and P. ostreatus. Phytochemical composition revealed that the infusion fraction of P. eous is the most enriched fraction having high amount of phenols (13.03 μggallic acid equivalent/mg of extract), flavonoids (3.57 μg quercetin equivalent/mg of extract) and ascorbic acid (16.66 μg/mg of extract). Moreover, free radical scavenging potentials of the corresponding fractions of these mushrooms were tested by different in-vitro systems. Results demonstrated infusion had noticeable scavenging activity in concentration dependent manner while decoction exhibited moderate activity for all three species. Decoction did not show any activity in hydroxyl radical scavenging assay. Together, our findings provide a detailed data explaining beneficial therapeutic properties of aqueous fraction of these Pleurotus species. Additionally, it also can be concluded that in comparison with other studied mushroom, P. eous infusion is superior in terms of antioxidant efficacy. [ABSTRACT FROM AUTHOR]

The protozoan parasite Cryptosporidium parvum (CP) causes cryptosporidiosis, a diarrheal disease worldwide. Infection in immunocompetent hosts typically results in acute, self-limiting, or recurrent diarrhea. However, in immunocompromised individuals infection can cause fulminant diarrhea, extraintestinal manifestations, and death. To date, the mechanisms underlying CP-induced diarrheal pathogenesis are poorly understood. Diarrheal diseases most commonly involve increased secretion and/or decreased absorption of fluid and electrolytes. We and others have previously shown impaired chloride absorption in infectious diarrhea due to dysregulation of SLC26A3 [downregulated in adenoma (DRA)], the human intestinal apical membrane Cl–/HCO3– exchanger protein. However, there are no studies on the effects of CP infection on DRA activity. Therefore, we examined the expression and function of DRA in intestinal epithelial cells in response to CP infection in vitro and in vivo. CP infection (0.5 X 106 oocysts/well in 24-well plates, 24 h) of Caco-2 cell monolayers significantly decreased Cl–/HCO3– exchange activity (measured as DIDS-sensitive 125I uptake) as well as DRA mRNA and protein levels. Substantial downregulation of DRA mRNA and protein was also observed following CP infection ex vivo in mouse enteroid-derived monolayers and in vivo in the ileal and jejunal mucosa of C57BL/6 mice for 24 h. However, at 48 h after infection in vivo, the effects on DRA mRNA and protein were attenuated and at 5 days after infection DRA returned to normal levels. Our results suggest that impaired chloride absorption due to downregulation of DRA could be one of the contributing factors to CP-induced acute, self-limiting diarrhea in immunocompetent hosts. [ABSTRACT FROM AUTHOR]

Impaired absorption of electrolytes is a hallmark of diarrhea associated with inflammation or enteric infections. Intestinal epithelial luminal membrane NHE3 (Na+/H+ exchanger 3) and DRA (Down-Regulated in Adenoma; Cl-/HCO3- exchanger) play key roles in mediating electroneutral NaCl absorption. We have previously shown decreased NHE3 and DRA function in response to short-term infection with enteropathogenic E. coli (EPEC), a diarrheal pathogen. Recent studies have also shown substantial downregulation of DRA expression in a diarrheal model of infection with Citrobacter rodentium, the mouse counterpart of EPEC. Since our previous studies showed that the probiotic Lactobacillus acidophilus (LA) increased DRA and NHE3 function and expression and conferred protective effects in experimental colitis, we sought to evaluate the efficacy of LA in counteracting NHE3 and DRA inhibition and ameliorating diarrhea in a model of C. rodentium infection. FVB/N mice challenged with C. rodentium [1 x 109 colony-forming units (CFU)] with or without administration of live LA (3 x 109 CFU) were assessed for NHE3 and DRA mRNA and protein expression, mRNA levels of carbonic anhydrase, diarrheal phenotype (assessed by colonic weight-to-length ratio), myeloperoxidase activity, and proinflammatory cytokines. LA counteracted C. rodentium-induced inhibition of colonic DRA, NHE3, and carbonic anhydrase I and IV expression and attenuated diarrheal phenotype and MPO activity. Furthermore, LA completely blocked C. rodentium induction of IL-1β, IFN-γ, and CXCL1 mRNA and C. rodentiuminduced STAT3 phosphorylation. In conclusion, our data provide mechanistic insights into antidiarrheal effects of LA in a model of infectious diarrhea and colitis. [ABSTRACT FROM AUTHOR]

We recently demonstrated that capturing human monoclonal antibodies (hmAbs) using high affinity anti-human Fc (AHC) antibodies allows reliable characterization of antibody-antigen interactions. Here, we characterized six human Fc specific mouse monoclonal antibodies (mAbs) and compared their binding profiles with three previously characterized goat AHC polyclonal antibodies (pAbs), exhibiting properties of a good capture reagent. All six mouse AHC mAbs specifically bound with high affinity to the Fc region of hIgG1, hIgG2, hIgG4 and to 43 different hIgG variants, containing substitutions and/or mutations in the hinge and/or Fc region, that have been reported to exhibit modified antibody effector function and/or pharmacokinetics. Biacore sensor surfaces individually derivatized with mouse AHC mAbs exhibited >2.5-fold higher hIgG binding capacity compared to the three goat AHC pAb surfaces and reproducibly captured hIgG over 300 capture-regeneration cycles. The results of the capture kinetic analyses performed on 31 antibody-antigen interactions using surfaces derivatized with either of the two highest affinity AHC mAbs (REGN7942 or REGN7943) were in concordance with those performed using goat AHC pAb surfaces. Our data demonstrate that AHC mAbs such as REGN7942 and REGN7943 that have properties superior than the three goat AHC pAbs are highly valuable research reagents, especially to perform capture kinetic analyses of antibody-antigen interactions on optical biosensors. [Display omitted] • All six mouse AHC mAb immobilized surfaces reproducibly captured hIgG even after 300 capture-regeneration cycles • Unlike the three goat AHC pAbs, the two highest affinity AHC mAbs (REGN7942 or REGN7943) stably captured even 500RU capture density of hIgG once immobilized on a C1 chip, exhibiting far superior properties as an AHC capture reagent • Capture kinetic analyses performed on 31 antibody-antigen interactions using surfaces derivatized with either REGN7942 or REGN7943 yielded results that were in concordance with those performed using the goat AHC pAb surfaces. [ABSTRACT FROM AUTHOR]

Rationale:Induced pluripotent stem (iPS) cells have emerged as a source of potentially unlimited supply of autologous endothelial cells (ECs) for vascularization. However, the regenerative function of these cells relative to adult ECs and ECs derived from embryonic stem (ES) cells is unknown. The objective was to define the differentiation characteristics and vascularization potential of Fetal liver kinase (Flk)1+ and Vascular Endothelial (VE)-cadherin+ ECs derived identically from mouse (m)ES and miPS cells. Methods and Results:Naive mES and miPS cells cultured in type IV collagen (IV Col) in defined media for 5 days induced the formation of adherent cell populations, which demonstrated similar expression of Flk1 and VE-cadherin and the emergence of EC progenies. FACS purification resulted in 100% Flk1+ VE-cadherin+ cells from both mES and miPS cells. Emergence of Flk1+VE-cadherin+ cells entailed expression of the vascular developmental transcription factor Er71, which bound identically to Flk1, VE-cadherin, and CD31 promoters in both populations. Immunostaining with anti-VE-cadherin and anti-CD31 antibodies and microscopy demonstrated the endothelial nature of these cells. Each cell population (unlike mature ECs) organized into well-developed vascular structures invitro and incorporated into CD31+ neovessels in matrigel plugs implanted in nude mice invivo. Conclusion:Thus, iPS cell-derived Flk1+VE-cadherin+ cells expressing the Er71 are as angiogenic as mES cell-derived cells and incorporate into CD31+ neovessels. Their vessel forming capacity highlights the potential of autologous iPS cells-derived EC progeny for therapeutic angiogenesis. [ABSTRACT FROM AUTHOR]