Neutrophil extracellular trap formation and citrullination in bronchiectasis

PhD ThesisBronchiectasis (BR) patients show risk for developing rheumatoid arthritis (BROS). As citrullination is implicated in rheumatoid arthritis pathogenesis it is possible that neutrophil extracellular trap (NET) formation, which is associated with PAD enzyme activity (i.e. citrullination), may be the mechanism connecting the two diseases. This body of work took 4 separate approaches to study the process of NETosis/PAD activity in the context of BR. Firstly, healthy peripheral blood neutrophils were assayed for changes in NETosis/PAD following stimulation with BR relevant molecules. Both NETosis and PAD activity increased following incubation, suggesting BR related stimuli promote NETosis and citrullination. Secondly, LPS signalling was assessed to determine how inhibition of the two arms of the signalling pathway (TRIF and MyD88) impacted NETosis/PAD activity. Pre-treatment with either inhibitor downregulated both NETosis rates in vitro, suggesting molecular signalling underpinning NETosis is broader and more complex than predicted. Thirdly, the impact of Cl-amidine (a PAD inhibitor) on NETosis, PAD activity and neutrophil function was assessed in vitro. Cl-amidine was shown to significantly reduce NETosis and PAD activity in response to BR stimuli. Superoxide production and phagocytosis was also shown to be inhibited by Cl-amidine, suggesting PAD plays some role these aspects of neutrophil function. Finally, NETosis, PAD activity and neutrophil function was assessed in healthy, BR and BROS neutrophils. Several differences in these results were observed between the groups, however limited sample size and lack of age matched healthy comparators complicate results interpretation. These results imply that the molecular mechanisms underpinning NETosis/PAD activation is likely more complicated than previously suggested, with the results also indicating that PAD activity may play a role in neutrophil function. BR relevant stimuli appear to promote both NETosis and citrullination, with preliminary data suggesting there may be some difference in these processes between healthy, BR and BROS patients

ASSESSMENT OF WIC COST-CONTAINMENT PRACTICES; EXECUTIVE SUMMARY

The Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) provides both nutrition education and supplemental foods containing nutrients determined by nutritional research to be lacking in the diets of pregnant, breastfeeding, and postpartum women, infants, and children. State WIC agencies have implemented practices designed to reduce the cost of food packages containing these prescribed foods. For instance, one of the WIC program's primary cost-saving practices is negotiating rebate contracts with manufacturers of infant formula. Additional practices include limiting authorized vendors to stores with lower food prices; limiting approved brands, package sizes, forms, or prices; and negotiating rebates with food manufacturers or suppliers. There is concern that these practices may inadvertently counter the program's goal of providing supplemental foods and nutrition education. Based on a review of cost-containment practices in six States, including interviews with the various stakeholders and analysis of WIC administrative files, the study draws three major conclusions: (1) costcontainment practices reduced average food package costs by 0.2 to 21.4 percent, depending on practices implemented and local conditions; (2) the cost-containment practices had few adverse outcomes for WIC participants; and (3) administrative costs of the practices were low, averaging about 1.5 percent of food package savings. The full report, Assessment of WIC Cost-Containment Practices: Final Report, is available online at http://www.ers.usda.gov/publications/efan03005.WIC Program, cost-containment, food-item restrictions, vendor restrictions, manufacturers' rebates, food package costs, Food Consumption/Nutrition/Food Safety, Food Security and Poverty,

Effects of hunting and non-hunting mortality on seasonal bobwhite densities in Tennessee

This thesis delineates some of the effects of hunting and non-hunting mortality on bobwhite populations on 8 study areas in middle and eastern Tennessee. Bobwhite population levels were measured via direct counts prior to (fall) and following (spring) annual hunting seasons from 1987 through 1991. The objectives of the study were to measure selected seasonal population levels and annual changes, determine effects of hunting and non-hunting mortality on measured population parameters, and assess hunting effort exerted on an intensively hunted area. The principal hypothesis investigated was that hunting and non-hunting mortality affected seasonal population levels and annual changes differently. Fall densities on the 8 study areas ranged from 16 to 126 bobwhites per 100 ha, and spring densities varied from 0 to 46 per 100 ha. Spring densities were positively related (P \u3c 0.05) to fall densities. Fall densities directly affected (P \u3c 0.05) succeeding fall densities. Fall densities significantly (P \u3c 0.05) influenced losses over winter, but had no effect on both hunting and non-hunting mortality when these two types of mortality were partitioned and compared separately. Declines in fall populations over winter varied from 0.0% to 100% on exploited areas. Populations unexploited by hunting had over-winter reductions ranging from 37.3% to 93.2%; 1 nonhunted population increased over winter. Hunting mortality ranged from 3.4% to 93.9% of fall densities. Non-hunting mortality for exploited populations reduced fall densities 3.3% to 96.6%, while unexploited populations recorded non-hunting losses of 37.5% to 93.2%. Hunting and non-hunting mortality rates for exploited populations were not correlated. From 1988-89 through 1990-91, hunting mortality operated as a compensatory form of reduction, while in 1987-88, hunting was at least partially additive on one study area. Summer gains were inversely related to spring densities (P \u3c 0.05). Spring populations increased 5.3% to 441.5% over summer. Hunted populations increased over summer throughout the study. Non-hunted populations increased some years on some areas and decreased on others. No statistical relationship existed between hunting mortality and over-winter losses, summer gains, and fall-fall changes. Proportions removed by causes unrelated to hunting were highly correlated with spring densities, over-winter losses, and summer gains, but not correlated with fall-fall changes. On the intensively hunted Jones study area, total harvest, hunter success, and hunting pressure varied appreciably from year to year, but the average per trip afield for these variables differed little among years

The Role of DNMT3A in Acute Myeloid Leukemia Pathogenesis

Loss of function mutations in the DNA methyltransferase DNMT3A are highly recurrent in acute myeloid leukemia (AML). DNMT3A and the highly homologous gene DNMT3B encode the two methyltransferases that are primarily responsible for mediating de novo methylation of specific DNA sequences during cellular differentiation. DNMT3A mutations are mutually exclusive of several translocations that create oncogenic fusion genes (PML-RARA, RUNX1-RUNX1T1, CBFB-MYH11, and MLL-X), suggesting that these fusions may require functional DNMT3A to initiate leukemogenesis. Using bone marrow cells from a constitutive Dnmt3a null mouse, we show that loss of Dnmt3a caused a striking loss of DNA methylation throughout the genome of bone marrow cells, and a complete loss of methylation at hundreds of specific loci, suggesting that these regions are entirely dependent on Dnmt3a for maintaining normal methylation patterns. Using both retroviral vectors and a transgenic model, we demonstrated that the methyltransferase activity of Dnmt3a but not Dnmt3b is required for aberrant self-renewal ex vivo that is driven by PML-RARA (but not RUNX1-RUNX1T1 or MLL-AF9); further, the PML-RARA competitive transplant advantage and leukemia generation both required Dnmt3a. In contrast, Dnmt3a was not required for leukemia generation caused by MLL-AF9, which is known to have a requirement for Dnmt1 activity. Together, these findings demonstrate that PML-RARAs specifically dependent on Dnmt3a to drive leukemogenesis, and may explain why DNMT3A mutations are mutually exclusive of PML-RARA in AML patients. While the most common mutation in DNMT3A in AML patients is the missense mutation R882H, other heterozygous mutations produce frameshifts, premature stop codons, or deletions of the entire coding sequence of the gene, strongly suggesting that these mutations lead to simple haploinsufficiency for DNMT3A. To test the hypothesis that Dnmt3a haploinsuffiency may initiate AML, we performed a long-term tumor watch comparing wild-type mice (Dnmt3a+/+) to mice carrying one wild-type Dnmt3a allele and one targeted allele that contains a neomycin-resistance cassette inserted into the sequence coding for the catalytic domain of the protein, producing a true null allele (Dnmt3a+/- mice). At 6 weeks of age, Dnmt3a+/- mice have normal hematopoiesis, with no detected differences from wild-type littermates in myeloid, lymphoid, erythroid, or stem/progenitor populations in the bone marrow or spleen. However, after 1.5 years of age, 15/43 Dnmt3a+/- mice (35%) became moribund and were euthanized for pathologic evaluation, and at conclusion of the tumor watch at 2 years similar pathologic findings were observed in an additional 9 Dnmt3a+/- mice, for an overall disease penetrance of 24/43 (56%). In contrast, 0/20 WT littermate control animals developed myeloid malignancies over the same time period. Based on flow cytometric and morphologic findings, we classified 16 splenic tumors according to the Bethesda criteria: 11/16 had myeloid proliferative disease/MPD, 2/16 had myeloid leukemia with maturation, 2/16 had MPD-like myeloid leukemia, and 1 case had myeloid sarcoma. Six tumors out of 18 tested were able to successfully engraft and lead to lethal disease in sublethally irradiated wild-type recipients, providing further evidence that these tumors represent transplantable, cell-autonomous myeloid malignancies. Exome sequencing of engrafted tumors revealed mutations in the Ras/MAPK pathway, including the canonical gain-of-function mutation Kras G12C, a Ptpn11 E76K mutation, and a missense mutation in the tumor suppressor Neurofibromatosis 1 (Nf1). Importantly, 9/51 AML samples with DNMT3A mutations in the TCGA AML cohort also contained activating NRAS or KRAS mutations. Examination of the Dnmt3a locus in 4 sequenced samples revealed no evidence for mutations in or deletions of the residual wild-type Dnmt3a allele. These data strongly suggest that Ras/MAPK pathway mutations can cooperate with Dnmt3a haploinsufficiency to induce AML in C57Bl/6 mice and in humans.

Conceptualizing Positive versus Negative Awe: Do Connection and Self-Significance Interact to Determine Awe’s Valence?

In this dissertation, I adopted an appraisal theory approach to differentiating positive and negative experiences of awe. In addition to assessing traditional appraisal dimensions, I focused on self-diminishment and connectedness as the appraisals hypothesized to best differentiate awe by valence. I predicted that self-diminishment and connectedness would interact to determine whether awe is experienced as positive or negative, arguing that feeling “small” can be positive if paired with feeling connected but that feeling small can be negative if paired with feeling isolated. An exploratory study (n = 742) induced participants to feel an emotion (positive awe, negative awe, joy, or fear) before rating the descriptiveness of 24 appraisals of the emotion-inducing experience (including self-diminishment and connectedness). Compared to positive awe, negative awe was associated with greater appraisals of need for accommodation, self-diminishment, and isolation, and lower appraisals of connectedness and certainty. Five pilot studies were conducted to validate a video awe induction of awe and essay-based manipulations of self-diminishment and connectedness manipulations. Finally, Experiment 2 (n = 309) tested the theorized model that self-significance and connectedness interact to differentiate positive and negative awe; Experiment 2 also included an exploratory eye-tracking sample (n = 62). Although the predicted interaction was not significant, connectedness was associated with experiencing awe as more positive and less negative. Additional analyses suggested that whether one feels connected or isolated may change whether feelings of significance are experienced positively or negatively. Together, the studies are a further step in using appraisal theory to understand awe’s variants

Modelling the effect of sensory dynamics on a driver’s control of a nonlinear vehicle

In previous work a linear model of driver steering control was developed which takes account of human sensory dynamics and limitations. In this paper various approaches to modelling a driver’s control of a nonlinear vehicle are compared. In contrast to research focussed on modelling the optimal driver, the aim of this work is to develop a realistic model of driver steering behaviour. Simulations were run to compare various nonlinear state estimators and controllers. In general a trade-off was found between simulation time, which could also represent mental load, and controller performance. Experiments are planned to compare the results of these simulations against measured steering behaviour from human drivers

Intervention in the Tax Court and the Appellate Review of Tax Court Procedural Decisions

The Tax Court is an Article I court. It resolves more than 95% of all tax-related litigation—actually nearly 97% of the total federal tax docket in 2012. Despite this substantial role in federal litigation, scholars and courts have generally put aside the issue of what standard is appropriate when a U.S. federal court of appeals reviews Tax Court procedural questions. Section 7482 of the Internal Revenue Code (I.R.C.) grants jurisdiction to the courts of appeals to review Tax Court decisions “in the same manner and to the same extent as decisions of the district courts in civil actions tried without a jury.”Unlike district courts, which must follow the Federal Rules of Civil Procedure (FRCP), the Tax Court operates under its own separate set of procedural rules—the Tax Court Rules of Practice and Procedure (Tax Court Rules). Congress explicitly authorized the Tax Court to choose its own procedural rules under I.R.C. § 7453. Where there is no applicable Tax Court procedural rule, Tax Court Rule 1(b) provides that the Tax Court “may prescribe the procedure, giving particular weight to the Federal Rules of Civil Procedure to the extent that they are suitably adaptable to govern the matter at hand.”Currently, no judicial decision analyzes the appropriate standard for appellate review of a Tax Court’s decision to apply (or not apply) a federal procedural rule where the Tax Court lacks its own applicable procedure. The appropriate standard of appellate review for Tax Court procedural decisions garnered attention when the U.S. Virgin Islands (USVI) recently sought to intervene in several Tax Court proceedings under FRCP 24(a)(2). The Tax Court Rules lack an intervention procedure and the Tax Court, acting pursuant to its discretion under Rule 1(b), has never granted a third party non-taxpayer’s motion to intervene under FRCP 24(a)(2).Given the outcome-determinative potential of the standard of review,these recent USVI cases—discussed individually below— provide an important opportunity to determine whether the abuse of discretion or de novo standard should govern appellate review of Tax Court procedural decisions. In 2014, the U.S. Court of Appeals for the Eleventh Circuit became the third federal appellate court to overturn a Tax Court decision denying a motion by the USVI to intervene in a dispute between the Internal Revenue Service (IRS) and a taxpayer. In Huff v. Commissioner,10 the Eleventh Circuit joined the U.S. Courts of Appeals for the Third and Eighth Circuits in allowing the USVI to intervene in a Tax Court proceeding. However, not all federal appellate courts have ruled this way. The U.S. Court of Appeals for the Fourth Circuit created a circuit split when it affirmed the Tax Court in denying intervention to the USVI. The Huff decision deepened the current circuit split and complicatedthe matter further. The Eleventh Circuit in Huff—unlike the other three circuits that have addressed this issue—allowed intervention as a matter of right under FRCP 24(a)(2). This approach is novel, as the Tax Court has never allowed a third party non-taxpayer to intervene under this provision—although it has the discretion to do so under Tax Court Rule 1(b). The Huff decision presents two issues worthy of discussion in this Comment. The first is whether de novo review is appropriate when a court of appeals reviews Tax Court procedural decisions. Generally, under I.R.C. § 7482, the appeals courts review Tax Court decisions “in the same manner and to the same extent” as district court decisions. Yet, given the Tax Court’s general power to prescribe its own procedural rules under I.R.C. § 7453,16 the Huff court erred in applying the de novo standard of review to the Tax Court’s decision to not apply FRCP 24(a)(2). The second issue the Huff opinion presents—regarding the substance of the case—is whether the USVI has a right to intervene under FRCP 24(a)(2). This issue turns on whether a Tax Court proceeding is the proper forum in which to confront the weighty concerns of fair implementation and coordination of two separate but interrelated taxing agencies—the IRS and the USVI’s Bureau of Internal Revenue (BIR). Part I of this Comment briefly summarizes the pertinent facts of Huff. Part II surveys the current circuit split on intervention in Tax Court proceedings. Part III analyzes the proper standard for review of the USVI’s motion to intervene under FRCP 24(a)(2), as well as the proper standard regarding all appellate review of Tax Court procedural decisions—a discussion that is largely absent from both scholarly works and court opinions. This Comment concludes in Part III with a critique of the Huff court’s FRCP 24(a)(2) analysis

Intervention in the Tax Court and the Appellate Review of Tax Court Procedural Decisions

The Tax Court is an Article I court. It resolves more than 95% of all tax-related litigation—actually nearly 97% of the total federal tax docket in 2012. Despite this substantial role in federal litigation, scholars and courts have generally put aside the issue of what standard is appropriate when a U.S. federal court of appeals reviews Tax Court procedural questions. Section 7482 of the Internal Revenue Code (I.R.C.) grants jurisdiction to the courts of appeals to review Tax Court decisions “in the same manner and to the same extent as decisions of the district courts in civil actions tried without a jury.”Unlike district courts, which must follow the Federal Rules of Civil Procedure (FRCP), the Tax Court operates under its own separate set of procedural rules—the Tax Court Rules of Practice and Procedure (Tax Court Rules). Congress explicitly authorized the Tax Court to choose its own procedural rules under I.R.C. § 7453. Where there is no applicable Tax Court procedural rule, Tax Court Rule 1(b) provides that the Tax Court “may prescribe the procedure, giving particular weight to the Federal Rules of Civil Procedure to the extent that they are suitably adaptable to govern the matter at hand.”Currently, no judicial decision analyzes the appropriate standard for appellate review of a Tax Court’s decision to apply (or not apply) a federal procedural rule where the Tax Court lacks its own applicable procedure. The appropriate standard of appellate review for Tax Court procedural decisions garnered attention when the U.S. Virgin Islands (USVI) recently sought to intervene in several Tax Court proceedings under FRCP 24(a)(2). The Tax Court Rules lack an intervention procedure and the Tax Court, acting pursuant to its discretion under Rule 1(b), has never granted a third party non-taxpayer’s motion to intervene under FRCP 24(a)(2).Given the outcome-determinative potential of the standard of review,these recent USVI cases—discussed individually below— provide an important opportunity to determine whether the abuse of discretion or de novo standard should govern appellate review of Tax Court procedural decisions. In 2014, the U.S. Court of Appeals for the Eleventh Circuit became the third federal appellate court to overturn a Tax Court decision denying a motion by the USVI to intervene in a dispute between the Internal Revenue Service (IRS) and a taxpayer. In Huff v. Commissioner,10 the Eleventh Circuit joined the U.S. Courts of Appeals for the Third and Eighth Circuits in allowing the USVI to intervene in a Tax Court proceeding. However, not all federal appellate courts have ruled this way. The U.S. Court of Appeals for the Fourth Circuit created a circuit split when it affirmed the Tax Court in denying intervention to the USVI. The Huff decision deepened the current circuit split and complicatedthe matter further. The Eleventh Circuit in Huff—unlike the other three circuits that have addressed this issue—allowed intervention as a matter of right under FRCP 24(a)(2). This approach is novel, as the Tax Court has never allowed a third party non-taxpayer to intervene under this provision—although it has the discretion to do so under Tax Court Rule 1(b). The Huff decision presents two issues worthy of discussion in this Comment. The first is whether de novo review is appropriate when a court of appeals reviews Tax Court procedural decisions. Generally, under I.R.C. § 7482, the appeals courts review Tax Court decisions “in the same manner and to the same extent” as district court decisions. Yet, given the Tax Court’s general power to prescribe its own procedural rules under I.R.C. § 7453,16 the Huff court erred in applying the de novo standard of review to the Tax Court’s decision to not apply FRCP 24(a)(2). The second issue the Huff opinion presents—regarding the substance of the case—is whether the USVI has a right to intervene under FRCP 24(a)(2). This issue turns on whether a Tax Court proceeding is the proper forum in which to confront the weighty concerns of fair implementation and coordination of two separate but interrelated taxing agencies—the IRS and the USVI’s Bureau of Internal Revenue (BIR). Part I of this Comment briefly summarizes the pertinent facts of Huff. Part II surveys the current circuit split on intervention in Tax Court proceedings. Part III analyzes the proper standard for review of the USVI’s motion to intervene under FRCP 24(a)(2), as well as the proper standard regarding all appellate review of Tax Court procedural decisions—a discussion that is largely absent from both scholarly works and court opinions. This Comment concludes in Part III with a critique of the Huff court’s FRCP 24(a)(2) analysis