Publications

The human metallothionein (MT) IB gene (hMT-IB) is located in a region of human DNA containing at least four tandemly arranged MT genes. As deduced from its sequence, hMT-IB is likely to encode a functional protein. However, the predicted amino acid sequence differed from the hMT-I amino acid sequence in four positions. Most remarkable was the presence of an additional cysteine. Like other MT genes, hMT-IB has at least two copies of the metal-responsive element upstream from the transcription initiation site. These elements probably are responsible for the metal responsiveness of the hMT-IB promoter, leading to inducible expression of fused heterologous genes. Unlike the hMT-IIA and hMT-IA genes described previously, which are expressed in many different cell types, a high level of expression of the endogenous hMT-IB gene could be detected only in human hepatoma and renal carcinoma cell lines. Therefore, this is the first MT gene described which exhibits tissue specificity of expression. This specificity is controlled by a cis-acting mechanism involving methylation, since incubation of nonexpressing cells with an inhibitor of DNA methylation led to activation of the hMT-IB gene. In support of this notion, we found that the 5' flanking region of the hMT-IB gene was highly methylated in HeLa cells, a nonexpressing cell type, but it was not methylated in a hepatoma (expressing) cell line.

The 5' region of the acetylcholinesterase gene from the electric ray Torpedo californica has been cloned and its cap site identified. The 5' untranslated region is divided into two exons where a small exon extending between bp -22 to -60 is alternatively spliced. Cap sites are defined at two positions, bp -138 and -143. Twenty-one base pairs 5' of the -143 cap site a repeating TATA sequence is found. Further upstream in the gene consensus sequences for Sp1, AP1, and AP2 factors are evident. The promoter region of the acetylcholinesterase gene enhances transcription of a luciferase reporter gene transfected into C2 myoblasts. However, increased transcription was not evident after C2 myoblasts were induced to form myotubes. Cotransfection of this construct with c-Jun (AP1) and AP2 expression vectors shows marked increases of transcription rates in HepG2 and C2 cells. Protein kinase A elicited regulation of expression is also evident in quail fibroblasts. In gel retardation experiments both recombinant c-Jun (AP1) and AP2 proteins bind to the appropriate Torpedo sequences. Cellular extracts from the Torpedo electric organ exhibit AP2 binding activity. Thus, although all facets of specific regulation expected upon differentiation of mammalian muscle cells were not evident, the 5'-flanking region from the Torpedo AChE gene contains consensus sequences and functional promoter elements typical of mammalian nerve and muscle systems.

Valid and reliable measures are needed to better understand the relationship between physical activity and crime. This paper provides a comprehensive psychometric evaluation of measures developed in the Safe and Fit Environments (SAFE) Study to assess a conceptual framework. In addition to assessing the basic psychometric properties of each measure (e.g., variable distributions [item/scale level], internal consistency reliability), this study formally examined the measurement validity and invariance of measures across four age groups using confirmatory factor analysis. The sample (n=2,173) included 336 Adolescents (aged 12-17), 532 Young adults (aged 18-39), 838 Middle Adults, and 467 Older Adults (aged 66+). The psychometric evaluation of (sub)scales showed consistent factorial validity and internal consistency reliability across the majority of the measures and across the four age groups. Specifically, 14 of the 17 measures subjected to a test of factorial validity displayed statistically and practically significant factor loadings and internal consistency values in the overall sample and across the age groups. The pattern of correlations for each (sub)scale with other (sub)scales/indexes largely did not exhibit redundancy across measures. The findings expanded upon the test-retest reliability evaluation reported in Patch et al. (2019) and suggest these measures can be used in predictive models.

Abstract Major urinary protein-urokinase type plasminogen activator (MUP-uPA) transgenic mice develop liver disease resembling that described for albumin promoter-uPA transgenic mice. We have developed the GFP-MUP-uPA mice, by crossing MUP-uPA mouse with transgenic C57/BL6 mouse expressing GFP. Hepatocyte fluorescence in this model is relatively weak. In contrast, GFP fluorescence of non-parenchymal cells such as stellate cells, Kupper cells and endotherial cells is very bright green more than parenchymal cells enabling the portal vein and sinusoid structure to be clearly visualized Huh7 liver cancer cells expressing red fluorescent protein (RFP) were injected in the spleen of MUP-uPA transgenic mice. By day 7, the Huh7-RFP celsl were visualized in the liver with the Olympus FV1000 conforcal microscope. The hepatoma cells grew around the sinusoids of the host liver. The liver-metastatic Huh7-RFP cells recruited bright GFP-expressing non-parenchymal cells. This model enables imaging the effects of uPA on liver metastasis and stromal recruitment enabling the metastasis to progress. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5533. doi:1538-7445.AM2012-5533

<h3>Objectives</h3> To describe the objectively measured physical activity characteristics of a diverse sample of sixth-grade girls, to examine influences on physical activity, and to report compliance with physical activity guidelines. <h3>Design</h3> Cross-sectional study. <h3>Setting</h3> Six locations across the United States. <h3>Participants</h3> A total of 1578 sixth-grade girls. Accelerometers were worn for 7 days, and data for 6 days were included in the analyses. <h3>Main Exposures</h3> Race/ethnicity, free or reduced-price lunch, and geographic region. <h3>Main Outcome Measures</h3> Six operational definitions of adequate activity (60 or 30 minutes of daily moderate to vigorous physical activity at or above 4.6, 3.8, or 3.0 metabolic equivalents) were used to examine whether girls met physical activity guidelines. <h3>Results</h3> Average times spent in sedentary, light, moderate, and vigorous activities were 460, 342, 18, and 6 min/d, respectively. White girls were more active than girls in other race/ethnic groups, and girls who did not receive free or reduced-price lunch were more active than girls who did. Girls in western states were most active. Percentages of girls in compliance with the 6 thresholds for adequate activity varied widely and ranged from 0.6% to 100.0%. <h3>Conclusions</h3> When physical activity is measured objectively and a 4.6–metabolic equivalents cut point for moderate to vigorous physical activity is used, most sixth-grade girls do not meet guidelines for adequate physical activity. One notable finding was the effect of different accelerometer scoring protocols on estimates of compliance. Conceptual and empirical work is needed to define appropriate physical activity for youth using objective physical activity measures.

Toll-like Receptor 3 (TLR3) is a pathogen pattern recognition receptor that plays a key role in innate immunity. TLR3 signalling has numerous functions in liver, both in health and disease. Here we report that TLR3 is expressed by quiescent hepatic stellate cells (HSC) where it functions to induce transcription and secretion of functional interferons as well as a number of other cytokines and chemokines. Upon transdifferentiation into myofibroblasts, HSCs rapidly loose the ability to produce interferon gamma (IFNγ). Mechanistically, this gene silencing may be due to Polycomb complex mediated repression via methylation of histone H3 lysine 27. In contrast to wild type, quiescent HSC isolated from tlr3 knockout mice do not produce IFNγ in response to Poly(I∶C) treatment. Therefore, quiescent HSC may contribute to induction of the hepatic innate immune system in response to injury or infection.

Background U.S. adolescents and young adults are using indoor tanning at high rates, even though it has been linked to both melanoma and squamous cell cancer. Because the availability of commercial indoor tanning facilities may influence use, data are needed on the number and density of such facilities. Methods In March 2006, commercial indoor tanning facilities in 116 large U.S. cities were identified, and the number and density (per 100,000 population) were computed for each city. Bivariate and multivariate analyses conducted in 2008 tested the association between tanning-facility density and selected geographic, climatologic, demographic, and legislative variables. Results Mean facility number and density across cities were 41.8 (SD=30.8) and 11.8 (SD=6.0), respectively. In multivariate analysis, cities with higher percentages of whites and lower ultraviolet (UV)index scores had significantly higher facility densities than those with lower percentages of whites and higher UV index scores. Conclusions These data indicate that commercial indoor tanning is widely available in the urban U.S., and this availability may help explain the high usage of indoor tanning.

ABSTRACT Pediatric non-alcoholic fatty liver disease (NAFLD) is escalating in the United States, with a limited mechanistic understanding. Triclosan (TCS) is a high-volume antimicrobial additive that has been detected in human breastmilk and shown in adult mice to cause hepatosteatosis. To examine the effect of TCS presented to neonatal mice through lactation, we exposed pregnant females to TCS in their diet and evaluated its impact on nursing neonates. TCS is efficiently transferred by lactation to newborn mice, causing significant fatty liver (FL) during the suckling period. Lactational delivery stimulated hepatosteatosis, triglyceride accumulation, endoplasmic reticulum (ER) stress, inflammation, and liver fibrosis. These events were mirrored by inhibition of key metabolic regulators, FGF21 and AMPK. De novo lipogenesis (DNL) induced by lactational TCS exposure was blocked in mice deficient in hepatic ATF4 . In primary hepatocytes, siRNA specific inhibition of PERK, an ATF4 upstream activator and initiator of ER stress, blocked TCS induced DNL. Also, in the absence of PPARα, which targets regulation of ATF4, TCS induced triglyceride accumulation and the induction of DNL was blocked. The administration of obeticholic acid (OCA), a potent FXR agonist, as well as activation of intestinal mucosal-regenerative gp130 signaling, led to reduced liver ATF4 expression, PPARα signaling, and DNL when neonates were exposed to TCS. In summary, TCS exposure via lactation leads to early indicators of NAFLD development accompanied by hepatosteatosis that were mediated in a PERK-eIF2α-ATF4-PPARα cascade. These studies indicate that mother to child transmission of environmental toxicants such as TCS may underlie the recent increases in pediatric NAFLD.

Parkinson's disease is characterized by accumulation of α-synuclein (αSyn). Release of oligomeric/fibrillar αSyn from damaged neurons may potentiate neuronal death in part via microglial activation. Heretofore, it remained unknown if oligomeric/fibrillar αSyn could activate the nucleotide-binding oligomerization domain (NOD)-like receptor (NLR) family pyrin domain-containing 3 (NLRP3) inflammasome in human microglia and whether anti-αSyn antibodies could prevent this effect. Here, we show that αSyn activates the NLRP3 inflammasome in human induced pluripotent stem cell (hiPSC)-derived microglia (hiMG) via dual stimulation involving Toll-like receptor 2 (TLR2) engagement and mitochondrial damage. In vitro, hiMG can be activated by mutant (A53T) αSyn secreted from hiPSC-derived A9-dopaminergic neurons. Surprisingly, αSyn-antibody complexes enhanced rather than suppressed inflammasome-mediated interleukin-1β (IL-1β) secretion, indicating these complexes are neuroinflammatory in a human context. A further increase in inflammation was observed with addition of oligomerized amyloid-β peptide (Aβ) and its cognate antibody. In vivo, engraftment of hiMG with αSyn in humanized mouse brain resulted in caspase-1 activation and neurotoxicity, which was exacerbated by αSyn antibody. These findings may have important implications for antibody therapies aimed at depleting misfolded/aggregated proteins from the human brain, as they may paradoxically trigger inflammation in human microglia.

Urban liveability is a global priority for creating healthy, sustainable cities. Measurement of policy-relevant spatial indicators of the built and natural environment supports city planning at all levels of government. Analysis of their spatial distribution within cities, and impacts on individuals and communities, is crucial to ensure planning decisions are effective and equitable. This paper outlines challenges and lessons from a 5-year collaborative research program, scaling up a software workflow for calculating a composite indicator of urban liveability for residential address points across Melbourne, to Australia's 21 largest cities, and further extension to 25 global cities in diverse contexts.