6 resultados para Labels.
em Duke University
Resumo:
Luminescent semiconductor nanocrystals, also known as quantum dots (QDs), have advanced the fields of molecular diagnostics and nanotherapeutics. Much of the initial progress for QDs in biology and medicine has focused on developing new biosensing formats to push the limit of detection sensitivity. Nevertheless, QDs can be more than passive bio-probes or labels for biological imaging and cellular studies. The high surface-to-volume ratio of QDs enables the construction of a "smart" multifunctional nanoplatform, where the QDs serve not only as an imaging agent but also a nanoscaffold catering for therapeutic and diagnostic (theranostic) modalities. This mini review highlights the emerging applications of functionalized QDs as fluorescence contrast agents for imaging or as nanoscale vehicles for delivery of therapeutics, with special attention paid to the promise and challenges towards QD-based theranostics.
Resumo:
BACKGROUND: Mechanical and in particular tactile allodynia is a hallmark of chronic pain in which innocuous touch becomes painful. Previous cholera toxin B (CTB)-based neural tracing experiments and electrophysiology studies had suggested that aberrant axon sprouting from touch sensory afferents into pain-processing laminae after injury is a possible anatomical substrate underlying mechanical allodynia. This hypothesis was later challenged by experiments using intra-axonal labeling of A-fiber neurons, as well as single-neuron labeling of electrophysiologically identified sensory neurons. However, no studies have used genetically labeled neurons to examine this issue, and most studies were performed on spinal but not trigeminal sensory neurons which are the relevant neurons for orofacial pain, where allodynia oftentimes plays a dominant clinical role. FINDINGS: We recently discovered that parvalbumin::Cre (Pv::Cre) labels two types of Aβ touch neurons in trigeminal ganglion. Using a Pv::CreER driver and a Cre-dependent reporter mouse, we specifically labeled these Aβ trigeminal touch afferents by timed taxomifen injection prior to inflammation or infraorbital nerve injury (ION transection). We then examined the peripheral and central projections of labeled axons into the brainstem caudalis nucleus after injuries vs controls. We found no evidence for ectopic sprouting of Pv::CreER labeled trigeminal Aβ axons into the superficial trigeminal noci-receptive laminae. Furthermore, there was also no evidence for peripheral sprouting. CONCLUSIONS: CreER-based labeling prior to injury precluded the issue of phenotypic changes of neurons after injury. Our results suggest that touch allodynia in chronic orofacial pain is unlikely caused by ectopic sprouting of Aβ trigeminal afferents.
Resumo:
*Designated as an exemplary master's project for 2015-16*
The American approach to disparities in educational achievement is deficit focused and based on false assumptions of equal educational opportunity and social mobility. The labels attached to children served by compensatory early childhood education programs have evolved, e.g., from “culturally deprived” into “at-risk” for school failure, yet remain rooted in deficit discourses and ideology. Drawing on multiple bodies of literature, this thesis analyzes the rhetoric of compensatory education as viewed through the conceptual lens of the deficit thinking paradigm, in which school failure is attributed to perceived genetic, cultural, or environmental deficiencies, rather than institutional and societal inequalities. With a focus on the evolution of deficit thinking, the thesis begins with late 19th century U.S. early childhood education as it set the stage for more than a century of compensatory education responses to the needs of children, inadequacies of immigrant and minority families, and threats to national security. Key educational research and publications on genetic-, cultural-, and environmental-deficits are aligned with trends in achievement gaps and compensatory education initiatives, beginning mid-20th century following the Brown vs Board declaration of 1954 and continuing to the present. This analysis then highlights patterns in the oppression, segregation, and disenfranchisement experienced by low-income and minority students, largely ignored within the mainstream compensatory education discourse. This thesis concludes with a heterodox analysis of how the deficit thinking paradigm is dependent on assumptions of equal educational opportunity and social mobility, which helps perpetuate the cycle of school failure amid larger social injustices.
Resumo:
Fibronectin (FN) is a large extracellular matrix (ECM) protein that is made up of
type I (FNI), type II (FNII), & type III (FNIII) domains. It assembles into an insoluble
supra-‐‑molecular structure: the fibrillar FN matrix. FN fibrillogenesis is a cell‐‑mediated process, which is initiated when FN binds to integrins on the cell surface. The FN matrix plays an important role in cell migration, proliferation, signaling & adhesion. Despite decades of research, the FN matrix is one of the least understood supra-‐‑molecular protein assemblies. There have been several attempts to elucidate the exact mechanism of matrix assembly resulting in significant progress in the field but it is still unclear as to what are FN-‐‑FN interactions, the nature of these interactions and the domains of FN that
are in contact with each other. FN matrix fibrils are elastic in nature. Two models have been proposed to explain the elasticity of the fibrils. The first model: the ‘domain unfolding’ model postulates that the unraveling of FNIII domains under tension explains fibril elasticity.
The second model relies on the conformational change of FN from compact to extended to explain fibril elasticity. FN contain 15 FNIII domains, each a 7-‐‑strand beta sandwich. Earlier work from our lab used the technique of labeling a buried Cys to study the ‘domain unfolding’ model. They used mutant FNs containing a buried Cys in a single FNIII domain and found that 6 of the 15 FNIII domains label in matrix fibrils. Domain unfolding due to tension, matrix associated conformational changes or spontaneous folding and unfolding are all possible explanation for labeling of the buried Cys. The present study also uses the technique of labeling a buried Cys to address whether it is spontaneous folding and unfolding that labels FNIII domains in cell culture. We used thiol reactive DTNB to measure the kinetics of labeling of buried Cys in eleven FN III domains over a wide range of urea concentrations (0-‐‑9M). The kinetics data were globally fit using Mathematica. The results are equivalent to those of H-‐‑D exchange, and
provide a comprehensive analysis of stability and unfolding/folding kinetics of each
domain. For two of the six domains spontaneous folding and unfolding is possibly the reason for labeling in cell culture. For the rest of the four domains it is probably matrix associated conformational changes or tension induced unfolding.
A long-‐‑standing debate in the protein-‐‑folding field is whether unfolding rate
constants or folding rate constants correlate to the stability of a protein. FNIII domains all have the same ß sandwich structure but very different stabilities and amino acid sequences. Our study analyzed the kinetics of unfolding and folding and stabilities of eleven FNIII domains and our results show that folding rate constants for FNIII domains are relatively similar and the unfolding rates vary widely and correlate to stability. FN forms a fibrillar matrix and the FN-‐‑FN interactions during matrix fibril formation are not known. FNI 1-‐‑9 or the N-‐‑ terminal region is indispensible for matrix formation and its major binding partner has been shown to be FNIII 2. Earlier work from our lab, using FRET analysis showed that the interaction of FNI 1-‐‑9 with a destabilized FNIII 2 (missing the G strand, FNIII 2ΔG) reduces the FRET efficiency. This efficiency is restored in the presence of FUD (bacterial adhesion from S. pyogenes) that has been known to interact with FNI 1-‐‑9 via a tandem ß zipper. In the present study we
use FRET analysis and a series of deletion mutants of FNIII 2ΔG to study the shortest fragment of FNIII 2ΔG that is required to bind FNI 1-‐‑9. Our results presented here are qualitative and show that FNIII 2ΔC’EFG is the shortest fragment required to bind FNI 1-‐‑9. Deletion of one more strand abolishes the interaction with FNI 1-‐‑9.
Resumo:
What role does socialization play in the origins of prosocial behavior? We examined one potential socialization mechanism, parents' discourse about others' emotions with very young children in whom prosocial behavior is still nascent. Two studies are reported, one of sharing in 18- and 24-month-olds (n = 29), and one of instrumental and empathy-based helping in 18- and 30-month-olds (n = 62). In both studies, parents read age-appropriate picture books to their children and the content and structure of their emotion-related and internal state discourse were coded. Results showed that children who helped and shared more quickly and more often, especially in tasks that required more complex emotion understanding, had parents who more often asked them to label and explain the emotions depicted in the books. Moreover, it was parents' elicitation of children's talk about emotions rather than parents' own production of emotion labels and explanations that explained children's prosocial behavior, even after controlling for age. Thus, it is the quality, not the quantity, of parents' talk about emotions with their toddlers that matters for early prosocial behavior.
