Adolescent Perceptions of Family Connectedness and School Belonging: Links with Self-Concept and Depressive Symptoms among Gifted African American and Hispanic Youth

Gifted minority students, particularly those who are African-American or Hispanic, often face significant barriers to their optimal psychosocial functioning and academic achievement. Lack of access to appropriate educational resources, reduced teacher expectations, under-identification and underrepresentation in gifted and talented programs, and outright discrimination all contribute to increased risk and reduced psychosocial and academic functioning among these youth. It is clear from the present results that sources of attachment are important for healthy functioning in gifted African American and Hispanic youth. Given the lack of presence of these students in gifted and talented programs nationwide, and the difficulty in retaining these students once enrolled, researchers and practitioners face increasing need to continue to strengthen the “family-school-community” link in order to enhance resiliency and reduce risk in gifted African American and Hispanic youth

the role of firms

Der Standortwechsel von Firmen in Länder mit niedrigen Sozialstandards wird in der Regel als treibende Kraft des abwärtsgerichteten regulativen Wettbewerbs zwischen Staaten angesehen. Es gibt allerdings eine Vielzahl von Fällen in denen genau das Gegenteil passiert: Firmen halten sich freiwillig an Sozialstandards (CSR) und üben sogar Druck auf Regierungen aus, um striktere Regulierung zu erwirken. In diesem Arbeitspapier zeigen wir, unter welchen Bedingungen Firmen zur Verwirklichung anspruchsvollerer Sozialstandards in Ländern mit geringer regulativer Kapazität beitragen. Zunächst stellen wir Hypothesen aus der bereits existierenden Literatur vor und arbeiten ihre Erklärungskraft für die hier diskutierte Problematik heraus. Das Arbeitspapier untersucht die Reaktion der südafrikanischen Textil- und Automobilindustrie auf die HIV Pandemie. Die südafrikanische Regierung hat nur begrenzte Kapazitäten aufgebracht, um sich gegen die Verbreitung des Virus zur Wehr zu setzen. Unter welchen Bedingungen versuchen Firmen den Staat im Kampf gegen HIV/AIDS zu unterstützen?Firms relocating production to countries with lower social standards are regarded as driving force behind the regulatory ‘race to the bottom’. However, there are numerous instances in which the behavior of firms reveals just the opposite: They adhere to self-regulatory standards (CSR) and even pressure governments to issue stricter public regulations. We intend to identify the conditions under which firms contribute to higher regulatory standards in states with weak regulatory capacities, thereby following a ‘race to the top’ rather than a ‘race to the bottom’- logic. Theoretically, we set out to test in how far the existing literature can be utilized to answer this question. Empirically, the assessment concentrates on the textile and automotive industries in South Africa and HIV/AIDS abatement. Only limited state capacities have been involved in fighting HIV/AIDS in South Africa. Under which conditions do firms try to foster state capacities for the fight against the disease

Codon Optimization for Alpha 1-Antitrypsin Disease

Alpha 1-antitrypsin deficiency is a genetic disorder caused by defective production of alpha 1-antitrypsin (AAT). Gene therapy approaches have been conducted in patients with AAT deficiency with successful AAT expression, but not to the therapeutic levels required to reduce the risk of emphysema. Codon optimization, a somewhat new and evolving technique, is used by many scientists to maximize protein expression in living organisms by altering translational and transcriptional efficiency as well as protein refolding. The purpose of this study was to develop single stranded and double stranded AAT gene constructs, test their protein expression in vitro, and compare with those levels expressed by the AAT construct that is currently in clinical trials. Three constructs were to be developed, yet only one construct was successfully cloned. This clone, optimized ds-CB-AAT, illustrated increased AAT protein expression as the transfection time increased. However, protein levels were appreciably lower in the optimized construct compared to the single stranded (long intron) AAT construct that is currently being administered in clinical trials. The data did not suggest that the optimized AAT construct does in fact express more AAT protein in vitro as expected. In order to achieve data that can be reproduced, the 2 remaining constructs need to be cloned and all of the isolated plasmid DNA should be prepared on the same scale to minimize any additional confounding variables

Hepatic Changes Associated with Chronic Alcohol Exposure in an Alpha-1 Antitrypsin PiZ Mouse Model

The PiZ mutation in the alpha-1 antitrypsin (AAT) gene causes the PiZ mutant protein to be sequestered in the endoplasmic reticulum of hepatocytes, causing significant liver pathology in ~10% of PiZZ homozygous AAT disease patients. Current transgenic mouse models of the disease include the liver-specific over-expression of mutant PiZ protein. However, these animal models do not efficiently recapitulate the liver damage found in PiZZ homozygous patients. Since only a small percentage of patients develop liver disease and it is not reproducible in animal models of AATD, it suggests that there are other factors that participate in disease pathogenesis. Here, we propose that in the presence of alcohol, liver injury will be initiated and that the intensity of the disease will be exacerbated by the presence of accumulated PiZ mutant protein. To test this hypothesis, we have administered alcohol via the Lieber-DeCarli diet regimen to PiZ transgenic and control C57Bl/6 mice for 12 weeks. We found no difference in alcohol and non-alcohol fed mice in terms of elevations in liver enzymes (AST and ALT). We did find a difference in the degree of steatosis and inflammation in the livers of alcohol fed PiZ mice over those of control alcohol fed mice. These findings are consistent with a chronic low-level hepatic insult seen in chronic alcohol consumption. The difference between PiZ and control mice will allow us to test gene therapies that prevent the accumulation of PiZ aggregates within hepatocytes to determine if they will prevent the exacerbation of alcoholic liver disease

Muscle-Directed Delivery of an AAV1 Vector Leads to Capsid-Specific T Cell Exhaustion in Nonhuman Primates and Humans

With the US Food and Drug Administration (FDA) and European Medicines Agency (EMA) approvals for Zolgensma, Luxturna, and Glybera, recombinant adeno-associated viruses (rAAVs) are considered efficient tools for gene transfer. However, studies in animals and humans demonstrate that intramuscular (IM) AAV delivery can trigger immune responses to AAV capsids and/or transgenes. IM delivery of rAAV1 in humans has also been described to induce tolerance to rAAV characterized by the presence of capsid-specific regulatory T cells (Tregs) in periphery. To understand mechanisms responsible for tolerance and parameters involved, we tested 3 muscle-directed administration routes in rhesus monkeys: IM delivery, venous limb perfusion, and the intra-arterial push and dwell method. These 3 methods were well tolerated and led to transgene expression. Interestingly, gene transfer in muscle led to Tregs and exhausted T cell infiltrates in situ at both day 21 and day 60 post-injection. In human samples, an in-depth analysis of the functionality of these cells demonstrates that capsid-specific exhausted T cells are detected after at least 5 years post-vector delivery and that the exhaustion can be reversed by blocking the checkpoint pathway. Overall, our study shows that persisting transgene expression after gene transfer in muscle is mediated by Tregs and exhausted T cells

rAAV9 airway delivery results in effective knockdown of mutant alpha 1-antitrypsin in the liver while upregulating wildtype alpha 1-antitrypsin in the lung

Alpha 1-Antitrypsin (AAT) deficiency is a human genetic disease resulting in the production of mutant AAT, a hepatocyte produced serine protease inhibitor that functions to prevent alveolar epithelial damage by inhibiting neutrophil elastase. Patients with AAT deficiency have increased lung disease, due to decreased proteolytic protection, as well as sporadic severe liver disease secondary to accumulation of mutant AAT, especially a common mutant form termed PiZ, within hepatocytes. We previously showed, in a PiZ mutant mouse model, simultaneous knock-down of mutant PiZ-AAT and augmentation of wild-type AAT production through intravenous delivery of a recombinant adeno-associated viral (rAAV) vector encoding both a miRNA targeting PiZ-AAT and a miRNA-resistant wild-type AAT gene. In this study we tested the hypothesis that rAAV2/9 vector administered intra-nasally or intra-tracheally can deliver a gene of interest to both the airways and liver. Initially C57Bl/6 mice were administered intra-nasally 1011 genome copies (GC) of rAAV2/9 vector expressing a firefly luciferase, which resulted in increased luminescence in the nasal passages, liver, and lung 21 days post delivery. Next, 1012 GC of rAAV2/9 vector expressing GFP and miRNAs targeting PiZ-AAT were delivered via oro-tracheal intubation to PiZ mice. This resulted in decreased serum AAT levels in the PiZ mice and GFP expression in both the liver and lungs. Finally, 1012 GC of rAAV2/9 vector encoding miRNA resistant wild-type AAT and miRNAs targeting PiZ-AAT were delivered via oro-tracheal intubation. This resulted in both systemic and local (liver and lung) elevations in wild-type AAT as well as decreased PiZ-AAT levels. In conclusion, tracheal delivery of rAAV2/9 resulted in expression of AAT in the liver and lung of treated animals, with sufficient targeting of the liver to mediate knock-down of mutant AAT to a similar degree as intravenous delivery, representing a potential non-invasive delivery route for gene therapy in AAT deficient patients

Thermal regime of the Costa Rican convergent margin: 1. Along-strike variations in heat flow from probe measurements and estimated from bottom-simulating reflectors

21 pages, 9 figures, 1 tableThe thermal structure of convergent margins provides information related to the tectonics, geodynamics, metamorphism, and fluid flow of active plate boundaries. We report 176 heat flow measurements made with a violin bow style probe across the Costa Rican margin at the Middle America Trench. The probe measurements are collocated with seismic reflection lines. These seismic reflection lines show widespread distribution of bottom-simulating reflectors (BSRs). To extend the spatial coverage of heat flow measurements we estimate heat flow from the depth of BSRs. Comparisons between probe measurements and BSR-derived estimates of heat flow are generally within 10% and improve with distance landward of the deformation front. Together, these determinations provide new information on the thermal regime of this margin. Consistent with previous studies, the margin associated with the northern Nicoya Peninsula is remarkably cool. We define better the southern boundary of the cool region. The northern extent of the cool region remains poorly determined. A regional trend of decreasing heat flow landward of the deformation front is apparent, consistent with the downward advection of heat by the subducting Cocos Plate. High wave number variability at a scale of 5–10 km is significantly greater than the measurement uncertainty and is greater south of the northern Nicoya Peninsula. These heat flow anomalies vary between approximately 20 and 60 mW m−2 and are most likely due to localized fluid flow through mounds and faults on the margin. Simple one-dimensional models show that these anomalies are consistent with flow rates of 7–15 mm yr−1. Across the margin toe variability is significant and likely due to fluid flow through deformation structures associated with the frontal sedimentary prismThis research was support by an NSF award (OCE‐0637120) to R.N.H. We thank R. von Huene, P. Fulton, and G. Spinelli for helpful comments. Heat flow data acquisition was funded by the German Science Foundation (DFG) through grant Vi 133/7‐1 to H.V. and I.G. and the SFB 574 “Volatiles and fluids in subduction zones” at Christan‐Albrechts University, Kiel. This is a contribution of the Barcelona Center for Subsurface Imaging (Barcelona‐CSI) supported by the Kaleidoscope project of REPSOLPeer Reviewe

Serum Levels of Alpha-1 Antitrypsin following Vascular Limb or Intra-Muscular Delivery of AAV1 or AAV8 Gene Therapy Vectors in Rhesus Macaques

Alpha-one antitrypsin (AAT) deficiency is a genetic disease that results in both lung disease and potentially liver failure in affected patients. In un-affected people AAT is produced in the liver and secreted to act as an anti-protease (primarily counteracting the effects of neutrophil elastase) in the lung. On-going human clinical trials have focused on intra-muscular delivery of adeno-associated virus (AAV1) to patients. The goal of delivery to the muscle is to have the myocytes serve as bio-factories to produce normal AAT protein and secrete it into the blood where it can exert its normal function in the lung. In the last Phase II trial patients in the highest dose cohort were given 100 intra-muscular (IM) injections with serum AAT levels still below therapeutic thresholds. Previous work has shown that delivering AAV vector to the musculature of the limb via the vasculature, while blood flow is obstructed using a tourniquet, leads to wide-spread gene expression in myocytes. We hypothesize that local delivery via IM injection results in saturated AAT expression within the myocytes surrounding the injection sight and that a more widespread delivery would result in an overall increase in serum AAT levels with the same dose of AAV gene therapy vector due to production by a larger overall number of myocytes. We have been able to show that we can attain similar or slightly higher (573.0 ng/ml versus 562.5 ng/nl) serum AAT levels using a vascular delivery method in rhesus macaques when compared to IM delivery. These results have been obtained using AAV1. Animals receiving either AAV1 or AAV8 show a decrease in muscle immune cell infiltrates following intra-vascular delivery versus IM delivery, which may improve long-term expression. Serum AAT data from animals dosed using AAV8, a serotype shown to better target muscle following vascular delivery, are currently being processed