Elucidating the role of BCL6 in helper T cell activation, proliferation, and differentiation

Indiana University-Purdue University Indianapolis (IUPUI)The transcriptional repressor BCL6 has been shown to be essential for the differentiation of germinal center (GC) B cells and follicular T helper (TFH) cells. The interaction of TFH and GC B cells is necessary for the development of high affinity antibodies specific for an invading pathogen. Germline BCL6-deficient mouse models limit our ability to study BCL6 function in T cells due to the strong inflammatory responses seen in these mice. To overcome this, our lab has developed a new BCL6 conditional knockout (cKO) mouse using the cre/lox system, wherein the zinc finger region of the BCL6 gene is flanked by loxP sites. Mating to a CD4-Cre mouse allowed us to study the effects of BCL6 loss specifically in T cells, without the confounding effects seen in germline knockout models. Using this cKO model, we have reaffirmed the necessity of BCL6 for TFH differentiation, including its role in sustained CXCR5 surface expression, a signature marker for TFH cells. This model also allowed us to recognize the role of BCL6 in promoting the expression of PD-1, another key surface marker for TFH cells. Without BCL6, CD4+ T cells cannot express PD-1 at the high levels seen on TFH cells. Our discovery of DNMT3b as a target for BCL6 suggests BCL6-deficient T cells have increased DNA methyltransferase activity at the PD-1 promoter. This data establishes a novel pathway for explaining how BCL6, a transcriptional repressor, can activate genes. Experiments with the BCL6 cKO model have also established a role for BCL6 in naïve CD4+ T cell activation. Furthermore, we did not observe increased differentiation of other helper T cell subsets, in contrast to what has been reported elsewhere with germline BCL6-deficient models. Unexpectedly, we found decreased T helper type 2 (Th2) cells, whereas mouse models with a germline mutation of BCL6 have increased Th2 cells. These results indicate that BCL6 activity in non-T cells is critical for controlling T cell differentiation. Finally, using an HIV-1 gp120 immunization model, we have, for the first time, shown BCL6-dependent GCs to be limiting for antibody development and affinity maturation in a prime-boost vaccine scheme

Exploration of factors behind the poor performance of low income men in Project Pioneer, a job training demonstration program in Maine

Thesis (M.C.P.)--Massachusetts Institute of Technology, Dept. of Urban Studies and Planning, 1998.Includes bibliographical references (p. 64).by Matissa N. Hollister.M.C.P

What do children who stutter and their parents expect from therapy and are their hopes aligned?

Currently there are few published reports exploring what it is that children and their parents wish to see change or happen as part of therapy. A process of eliciting hopes or expectations for therapy using Solution Focused Brief Therapy (de Shazer, 1985) is routinely used at the Michael Palin Centre (MPC) with children who stutter and their parents. This qualitative study aimed to explore what children who stutter and their parents expect from therapy and whether their hopes are aligned

The pore size of polycaprolactone scaffolds has limited influence on bone regeneration in an in vivo model

Bone tissue engineering scaffolds should be designed to optimize mass transport, cell migration, and mechanical integrity to facilitate and enhance new bone growth. Although many scaffold parameters could be modified to fulfill these requirements, pore size is an important scaffold characteristic that can be rigorously controlled with indirect solid freeform fabrication. We explored the effect of pore size on bone regeneration and scaffold mechanical properties using polycaprolactone (PCL) scaffolds designed with interconnected, cylindrical orthogonal pores. Three scaffold designs with unique microarchitectures were fabricated, having pore sizes of 350, 550, or 800 Μm. Bone morphogenetic protein-7 transduced human gingival fibroblasts were suspended in fibrin gel, seeded into scaffolds, and implanted subcutaneously in immuno-compromised mice for 4 or 8 weeks. We found that (1) modulus and peak stress of the scaffold/bone constructs depended on pore size and porosity at 4 weeks but not at 8 weeks, (2) bone growth inside pores depended on pore size at 4 weeks but not at 8 weeks, and (3) the length of implantation time had a limited effect on scaffold/bone construct properties. In conclusion, pore sizes between 350 and 800 Μm play a limited role in bone regeneration in this tissue engineering model. Therefore, it may be advantageous to explore the effects of other scaffold structural properties, such as pore shape, pore interconnectivity, or scaffold permeability, on bone regeneration when designing PCL scaffolds for bone tissue engineering. © 2009 Wiley Periodicals, Inc. J Biomed Mater Res, 2010Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/64538/1/32381_ftp.pd

Microwave Kinetic Inductance Detector (MKID) Camera Testing for Submillimeter Astronomy

Developing kilopixel focal planes for incoherent submm- and mm-wave detectors remains challenging due to either the large hardware overhead or the complexity of multiplexing standard detectors. Microwave kinetic inductance detectors (MKIDs) provide a efficient means to produce fully lithographic background-limited kilopixel focal planes. We are constructing an MKID-based camera for the Caltech Submillimeter Observatory with 576 spatial pixels each simultaneously sensitive in 4 bands at 230, 300, 350, and 400 GHz. The novelty of MKIDs has required us to develop new techniques for detector characterization. We have measured quasiparticle lifetimes and resonator Qs for detector bath temperatures between 200 mK and 400 mK. Equivalent lifetime measurements were made by coupling energy into the resonators either optically or by driving the third harmonic of the resonator. To determine optical loading, we use both lifetime and internal Q measurements, which range between 15,000 and 30,000 for our resonators. Spectral bandpass measurements confirm the placement of the 230 and 350 GHz bands. Additionally, beam maps measurements conform to expectations. The same device design has been characterized on both sapphire and silicon substrates, and for different detector geometries. We also report on the incorporation of new shielding to reduce detector sensitivity to local magnetic fields

Regulatory Considerations in the Design and Manufacturing of Implantable 3D‐Printed Medical Devices

Three‐dimensional (3D) printing, or additive manufacturing, technology has rapidly penetrated the medical device industry over the past several years, and innovative groups have harnessed it to create devices with unique composition, structure, and customizability. These distinctive capabilities afforded by 3D printing have introduced new regulatory challenges. The customizability of 3D‐printed devices introduces new complexities when drafting a design control model for FDA consideration of market approval. The customizability and unique build processes of 3D‐printed medical devices pose unique challenges in meeting regulatory standards related to the manufacturing quality assurance. Consistent material powder properties and optimal printing parameters such as build orientation and laser power must be addressed and communicated to the FDA to ensure a quality build. Postprinting considerations unique to 3D‐printed devices, such as cleaning, finishing and sterilization are also discussed. In this manuscript we illustrate how such regulatory hurdles can be navigated by discussing our experience with our group's 3D‐printed bioresorbable implantable device.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/115990/1/cts12315.pd

Habitat Characteristics of Northern Bobwhite Quail-Hunting Party Encounters: A Landscape Perspective

Landcover data and bobwhite hunting records were used to assess both hunter habitat preferences and the frequency of northern bobwhite encounters by hunting parties in relation to habitat composition during the 1994-1995 and 1995-1996 hunting seasons at the Joseph W. Jones Ecological Research Center in southern Georgia. Patterns of habitat use by hunters, and the frequency of bobwhite encounters varied within and between years, depending on habitat quality, food availability, and other factors. Landscape-scale analyses of standardized bobwhite covey densities (based on coveys pointed in the field) and habitat composition and configuration for the 1994-1995 hunting season revealed that bobwhite densities were: (1) positively associated with the overall percentage agriculture and food plot habitat (reaching a maximum at 30-35% agriculture); and (2) positively associated with edge complexity, and positively associated with agricultural mean patch size [reaching a maximum at 2-3 hectares (5-6 acres)]. Consequently, larger food plots may be more important for increasing bobwhite encounter rates than numerous very small food plots [ \u3c 0.1 hectares (0.25 acres)]. Results of this, and related ongoing studies, have important implications for both landscape design and multiple use resource management. activities in the context of northern bobwhite habitat management in southern upland pine forest ecosystems

Personalized scaffolding technologies for alveolar bone regenerative medicine

Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/149271/1/ocr12275.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/149271/2/ocr12275_am.pd

Modulating signaling networks by CRISPR/Cas9-mediated transposable element insertion

In a recent past, transposable elements (TEs) were referred to as selfish genetic components only capable of copying themselves with the aim of increasing the odds of being inherited. Nonetheless, TEs have been initially proposed as positive control elements acting in synergy with the host. Nowadays, it is well known that TE movement into host genome comprises an important evolutionary mechanism capable of increasing the adaptive fitness. As insights into TE functioning are increasing day to day, the manipulation of transposition has raised an interesting possibility of setting the host functions, although the lack of appropriate genome engineering tools has unpaved it. Fortunately, the emergence of genome editing technologies based on programmable nucleases, and especially the arrival of a multipurpose RNA-guided Cas9 endonuclease system, has made it possible to reconsider this challenge. For such purpose, a particular type of transposons referred to as miniature inverted-repeat transposable elements (MITEs) has shown a series of interesting characteristics for designing functional drivers. Here, recent insights into MITE elements and versatile RNA-guided CRISPR/Cas9 genome engineering system are given to understand how to deploy the potential of TEs for control of the host transcriptional activity.Fil: Vaschetto, Luis Maria Benjamin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Cátedra de Diversidad Animal I; Argentin