bluuebunny/crossref_metadata_2025 huggingface dataset:
Typological review of environmental performance metrics (with illustrative examples for oil spill response)
Authors: Seager Thomas P , Satterstrom F Kyle , Linkov Igor , Tuler Seth P , Kay Rebecca
Published: March 2010

An intensification of interest in environmental assessment during the last 2 decades has driven corporate efforts to better document environmental goals, improve environmental management systems, and increase awareness of the environmental and ecological effects of business operations. This trend has been motivated partly by regulatory requirements (such as the Toxics Release Inventory in the United States) and partly by the inclination of some large manufacturing firms to embrace a broader social and environmental mission characterized as “sustainability” or “ecoefficiency.” Moreover, the importance of measurable objectives in the US government has been recognized at least since the Government Performance Results Act of 1993, which was intended to both improve the efficiency of government and the confidence of the American public in government managers. However, in management of environmental crises—such as catastrophic oil or chemical spills—development of measurable performance standards has lagged. Consequently, government spill managers are unable to define success in terms that are easily communicated to public and other stakeholder groups, and they could be disadvantaged in their efforts to deploy response resources with maximum efficiency. In this paper, we present a typological review of environmental assessment measures and summarize some of the current practices and strategic goals among federal agencies with regard to oil and chemical spills. A general approach to organizing metrics for oil spill response, restoration, and recovery is also presented. The results could improve planning efforts and communication among different federal, state, and local agencies and public or stakeholder groups involved in spill management.
An integrated health risk assessment approach to the study of mining sites contaminated with arsenic and lead
Authors: Jasso-Pineda Yolanda , Espinosa-Reyes Guillermo , González-Mille Donají , Razo-Soto Israel , Carrizales Leticia , Torres-Dosal Arturo , Mejía-Saavedra Jesús , Monroy Marcos , Ize Ana Irina , Yarto Mario , Díaz-Barriga Fernando
Published: March 2010

In order to test the value of an integrated approach for the analysis of health risks at contaminated sites, an integrated health risk assessment in a mining area was performed following 3 steps: 1) Environmental monitoring of surface soil, 2) assessment of exposure to metals in children and native rodents, and 3) DNA damage evaluation (comet assay) in children and rodents. These aspects also were studied in less exposed populations. Our results in humans showed that children living in the most polluted area (Villa de la Paz, Mexico) had higher lead blood concentrations (geometric mean of 13.8 μg/dL) and urinary arsenic levels (geometric mean of 52.1 μg/g creatinine) compared to children living in a control area (Matehuala, Mexico; blood lead of 7.3 μg/dL; urinary arsenic of 16.8 μg/g creatinine). Furthermore, the exposed children also had increased DNA damage (tail moment mean in Villa de la Paz of 4.8 vs 3.9 in Matehuala; p < 0.05). Results in rodents were identical. Animals captured in the polluted area had higher levels of arsenic (geometric mean of 1.3 μg/g in liver and 1.8 μg/g in kidney), lead (0.2 μg/g in liver and 0.9 μg/g in kidney), and cadmium (0.8 μg/g in liver and 2.2 μg/g in kidney), and increased DNA damage (tail moment mean of 18.2) when compared to control animals (arsenic in liver of 0.08 μg/g and kidney of 0.1 μg/g; lead in liver of 0.06 μg/g and kidney of 0.3 μg/g; cadmium in liver of 0.06 μg/g and kidney of 0.6 μg/g; and tail moment of 14.2). With the data in children and rodents, the weight-of-evidence for health risks (in this case DNA damage) associated with metal exposure in Villa de la Paz was strengthened. Therefore, a remediation program was easier to justify, and a feasibility study at this site is under way.
Chronic exposure to polynuclear aromatic hydrocarbons in natal habitats leads to decreased equilibrium size, growth, and stability of pink salmon populations
Authors: Heintz Ron A
Published: March 2010

The immediate and delayed effects of embryonic exposure to low levels of polynuclear aromatic hydrocarbons (PAHs) have been shown to reduce survival to maturity by 50% in exposed pink salmon populations. This suggests that chronically exposed populations could be extirpated over relatively few generations, but the effect of density dependence on extirpation rate is unknown. This study examines the interaction of PAH effects and randomly varying density dependence on a simulated population. The simulation derives from 70 years of observations made on a single pink salmon population and toxicity studies involving a hatchery population in the same watershed. Results from simulations involving exposure of 100% of the population to effects consistent with an aqueous PAH concentration of 18 nL/L indicate an 80% decrease in population productivity and an 11% probability of extinction after 35 generations. In contrast, population growth rate declined by only 5%. Further decreases in survival relative to that of observed PAH effects rapidly increase the probability of extinction. Data from these simulations demonstrate that, at low levels of exposure, density dependence can compensate for reduced population size and buffer the population against extinction. However, if equilibrium size is depressed sufficiently, random environmental variation overcomes the buffering effect of density dependence and extinction probability increases. These data demonstrate that extinction probability and population size are more sensitive measures of population effects than growth rate for wild populations regulated by density dependence.
An ecological risk assessment for insecticides used in adult mosquito management
Authors: Davis Ryan S , Peterson Robert KD , Macedo Paula A
Published: March 2010

West Nile virus (WNV) has been a concern for people across the United States since the disease was initially observed in the summer of 1999. Since 1999, WNV has caused the largest arboviral encephalitis epidemic in US history. Vector control management programs have been intensively implemented to control mosquitoes that carry WNV. Our deterministic ecological risk assessment focused on 6 common mosquito adulticides used in vector control, including 3 pyrethroids (d-phenothrin, resmethrin, and permethrin), pyrethrins, and 2 organophosphates (malathion and naled). Piperonyl butoxide, a synergist for the pyrethroids, was also assessed. Both aquatic and terrestrial nontarget organisms were considered for acute and chronic exposures to the adulticides. Tier I exposure estimates were derived from ISCST3 and AERMOD for deposition and air concentrations affecting terrestrial organisms and PRZM-EXAMS for standard pond concentrations affecting aquatic organisms. Nontargets exposed to adulticides included small mammals, birds, as well as aquatic vertebrates and invertebrates in a pond subject to receiving the chemical via drift and runoff. Risk quotients were obtained by comparing exposures to toxic endpoints. All risk quotients were low indicating that risks to ecological receptors most likely were small.
Screening procedure to assess the impact of urban stormwater temperature to populations of brown trout in receiving water
Authors: Rossi Luca , Hari Renata E
Published: March 2010

The discharge of urban stormwater may cause a sudden temperature increase in receiving waters that may be harmful to fish and other aquatic organisms. A screening procedure is proposed with temperature thresholds for the runoff from roofs and roads as well as for the receiving water system to protect brown trout from thermal damage. The stormwater temperature is calculated on the basis of a simple thermodynamic estimate for different latitudes. Only receiving waters with maximum daily mean temperatures of 22 °C (T1) are considered potential habitats for brown trout. The maximum temperature for a 1-h exposure time with a safety margin for 100% survival is 25 °C (T2), the sudden temperature change at the beginning of a rain event must not exceed 7 °C (T3), and fish-egg development requires the daily maximum temperature in winter to be below 12 °C (T4). Examples of stormwater runoff from roof or road surfaces from Switzerland validate our approach within ±0.5 °C. Effects of runoff into receiving waters without detailed data can be predicted within ±0.8 °C. With the restriction by T1, T2 seems not to be an acute problem at Swiss latitudes. T3 could play a role, especially if a large amount of runoff is discharged in small and rather cool rivers and streams. Finally, T4 deserves more attention than hitherto given. The proposed procedure may be a useful tool for assessing the influence of urban stormwater on the temperature of the receiving waters, particularly with regard to predicting the thermal impacts of urban or suburban runoff to populations of brown trout.
The sediments of the venice lagoon (Italy) evaluated in a screening risk assessment approach: Part I—application of international sediment quality guidelines
Authors: Apitz Sabine E , Barbanti Andrea , Bocci Martina , Carlin Anna , Montobbio Laura , Bernstein Alberto Giulio
Published: March 2010

Although it generally is agreed on in the scientific community that an evaluation of sediment quality solely based on total contaminant levels should be regarded only as a 1st tier or 1 LOE in a WOE framework, not all regulatory frameworks follow this approach. As will be discussed later, dredged material disposal in Venice Lagoon currently is regulated based upon the use of regionally developed SQGs as strict pass/fail criteria. The implications of this policy upon the management of the lagoon are under investigation, but studies on contaminant mobility and bioavailability and on sediment-related effects on biota and exposure (bioassays, biomarkers, bioaccumulation, biomagnification) having the characteristics (duration, number of sites and samples, frequency, number of chemicals and endpoints, etc.) to properly support (e.g., preliminary and/or detailed quantitative risk assessment) decisions on a basin scale have been carried out only recently in the Venice Lagoon (Thetis 2003; Losso et al. 2004; Carrer et al. 2005; Thetis 2005c).
The sediments of the venice lagoon (Italy) evaluated in a screening risk assessment approach: Part II—lagoon sediment quality compared to hot spots, regional, and international case studies
Authors: Apitz Sabine E , Barbanti Andrea , Bocci Martina , Carlin Anna , Montobbio Laura , Bernstein Alberto Giulio
Published: March 2010

The objective of this study was to carry out a critical comparison of data on the screening quality of surface sediments in Venice Lagoon (VL; main lagoon and its subbasins, Porto Marghera [PM], and Venice City Canals) and in other transitional and coastal ecosystems with various levels of human impact (urbanization and industrialization). To put VL in terms of reference and industrialized sites in the region, case studies were selected from the North Adriatic Region; to gain insight into how VL sediments compared to transitional areas throughout the world, case studies also were selected from a number of regions internationally. In order to compare regional levels of contamination, statistically processed sediment contaminant levels within a region (minimum, maximum, mean, and median), not individual sample values, are compared. The screening quality (relative to a variety of sediment quality guidelines) and the drivers of screening risk (based upon contaminant mixtures) of the VL sediments and other coastal and transitional sites are compared and discussed. The VL sediments have hazard quotients on the low end of the range typical of moderately urbanized and industrialized sites and higher than background conditions among the case studies reviewed. The Hg levels in the VL were generally higher than at other sites, and other contaminants were either equivalent or lower. Although sediments have somewhat higher levels of some contaminants and lower levels of other contaminants in PM and Venice City (VC) canals, levels for most contaminants are comparable to case studies with high levels of anthropogenic impact. For many contaminants of interest, PM (and for some, VC) sediments have some of the highest levels of any case study reviewed. How PM and VC rank when compared to other highly industrialized sites depends upon how data are synthesized and how ranges are taken into account. Actual risk must be evaluated using a weight-of-evidence approach, because natural background levels and site-specific bioavailability will differ both regionally and internationally.
The Exxon Valdez oil spill revisited and the dangers of normative science
Authors: Landis Wayne G
Published: March 2010

In the July 2006 issue of Integrated Environmental Assessment and Management, a paper by Harwell and Gentile was published assessing the present ecological significance of the impacts from the Exxon Valdez oil spill (EVOS). First, this paper compares the major conclusions of Harwell and Gentile and a paper reviewing the current impacts of EVOS by Peterson et al. as published by Science in 2003. Stark differences exist between the conclusions of the 2 papers regarding continuing impacts. Part of the difference appears to be the infusion of different social values or policy goals into each. Normative science is the use or interpretation of data in support of specific values or policies. Examples of values or policies intertwined with science are constructs such as ecosystem health, ecosystem integrity, ecological significance, and recovery. Examination of the environmental risk assessment and toxicology literature reveals that the symptoms of normative science are common and the implications widespread. Separation of science from policy or at a minimum a transparent acknowledgment of the science–policy interaction is clearly necessary in order to obtain a clear picture of the ecological system under investigation.
Separating the wheat from the chaff: The effective use of mathematical models as decision tools
Authors: Glaser David , Bridges Todd S
Published: March 2010

The purpose of this paper is to discuss the effective use of quantitative modeling in environmental decision making, with a particular focus on problems of contaminated sediment and surface water. The intended audience includes both model developers and model users. Our goal is to facilitate more effective communication among model developers and those using the information produced by models to aid decision making. We provide a series of observations or conclusions we have reached in our experience that are as follows. A model is a tool for evaluating alternate hypotheses; a model itself is not a hypothesis. All decisions are actually based upon models, either explicitly or implicitly. Models are used to address diagnostic and prognostic questions. Models can provide value added when applied throughout the lifetime of a project. Uncertainty, and therefore the need for models, is greater in systems near background. Models can provide useful information even when based on relatively small data sets. The utility of a model depends on the strength of the constraints placed upon it. The calibration process can be only partially specified a priori. Model calibration and evaluation require multiple lines of evidence. Uncertainty analysis is both qualitative and quantitative. Validation is provided by the application of the model under a wide range of conditions. Communication of the strength of model constraints is critical to model acceptance. We conclude that while models are often used in the evaluation of contaminated sediment problems, distrust in the use of models remains strong. The assessment of uncertainty is the factor most limiting acceptability.
Population-scale assessment endpoints in ecological risk assessment part II: Selection of assessment endpoint attributes
Authors: Landis Wayne G , Kaminski Laurel A
Published: March 2010

Because ecological services often are tied to specific species, the risk to populations is a critical endpoint and important feature of ecological risk assessments. In Part 1 of this series it was demonstrated that population scale assessment endpoints are important expressions of the valued components of ecological structures. This commentary reviews several of the characteristics of populations that can be evaluated and used in population scale risk assessments. Two attributes are evaluated as promising. The 1st attribute is the change in potential productivity of the population over a specified time period. The 2nd attribute is the change in the age structure of a population, expressed graphically or as a normalized effects vector (NEV). The NEV is a description of the change in age structure due to a toxicant or other stressor and appears to be characteristic of specific stressor effects.
Prepubertal exposure to cow's milk reduces susceptibility to carcinogen‐induced mammary tumorigenesis in rats
Authors: Nielsen Tina S. , Khan Galam , Davis Jennifer , Michels Karin B. , Hilakivi‐Clarke Leena
Published: March 2010

Cow's milk contains high levels of estrogens, progesterone and insulin‐like growth factor 1 (IGF‐1), all of which are associated with breast cancer. We investigated whether prepubertal milk exposure affects mammary gland development and carcinogenesis in rats. Sprague–Dawley rats were given either whole milk or tap water to drink from postnatal day (PND) 14 to PND 35, and thereafter normal tap water. Mammary tumorigenesis was induced by administering 7,12‐dimethylbenz[a]anthracene on PND 50. Milk exposure increased circulating E2 levels on PND 25 by 10‐fold (p < 0.001) and accelerated vaginal opening, which marks puberty onset, by 2.5 days (p < 0.001). However, rats exposed to milk before puberty exhibited reduced carcinogen‐induced mammary carcinogenesis; that is, their tumor latency was longer (p < 0.03) and incidence was lower (p < 0.05) than in the controls. On PND 25 and 50, mammary glands of the milk‐exposed rats had significantly less terminal end buds (TEBs) than the tap water‐exposed controls (p < 0.019). ER‐α protein levels were elevated in the TEBs and lobules of milk rats, compared to rats given tap water (p < 0.019), but no changes in cyclin D1 expression, cell proliferation or apoptosis were seen. IGF‐1 mRNA levels were reduced on PND 50 in the mammary glands of rats exposed to milk at puberty. Our results suggest that drinking milk before puberty reduces later risk of developing mammary cancer in rats. This might be mediated by a reduction in the number of TEBs and lower expression of IGF‐1 mRNA in the mammary glands of milk‐exposed animals.
Menopausal hormone therapy and breast cancer risk: Impact of different treatments. The European Prospective Investigation into Cancer and Nutrition
Authors: Bakken Kjersti , Fournier Agnès , Lund Eiliv , Waaseth Marit , Dumeaux Vanessa , Clavel‐Chapelon Françoise , Fabre Alban , Hémon Bertrand , Rinaldi Sabina , Chajes Véronique , Slimani Nadia , Allen Naomi E. , Reeves Gillian K. , Bingham Sheila , Khaw Kay‐Tee , Olsen Anja , Tjønneland Anne , Rodriguez Laudina , Sánchez Maria‐José , Etxezarreta Pilar Amiano , Ardanaz Eva , Tormo Maria‐José , Peeters Petra H. , van Gils Carla H. , Steffen Annika , Schulz Mandy , Chang‐Claude Jenny , Kaaks Rudolf , Tumino Rosario , Gallo Valentina , Norat Teresa , Riboli Elio , Panico Salvatore , Masala Giovanna , González Carlos A. , Berrino Franco
Published: March 2010

Menopausal hormone therapy (MHT) is characterized by use of different constituents, regimens and routes of administration. We investigated the association between the use of different types of MHT and breast cancer risk in the EPIC cohort study. The analysis is based on data from 133,744 postmenopausal women. Approximately 133,744 postmenopausal women contributed to this analysis. Information on MHT was derived from country‐specific self‐administered questionnaires with a single baseline assessment. Incident breast cancers were identified through population cancer registries or by active follow‐up (mean: 8.6 yr). Overall relative risks (RR) and 95% confidence interval (CI) were derived from country‐specific Cox proportional hazard models estimates. A total of 4312 primary breast cancers were diagnosed during 1,153,747 person‐years of follow‐up. Compared with MHT never users, breast cancer risk was higher among current users of estrogen only (RR: 1.42, 95% CI 1.23–1.64) and higher still among current users of combined MHT (RR: 1.77, 95% CI 1.40–2.24; p = 0.02 for combined vs. estrogen‐only). Continuous combined regimens conferred a 43% (95% CI: 19–72%) greater risk compared with sequential regimens. There was no significant difference between progesterone and testosterone derivatives in sequential regimens. There was no significant variation in risk linked to the estrogenic component of MHT, neither for oral vs. cutaneous administration nor for estradiol compounds vs. conjugated equine estrogens. Estrogen‐only and combined MHT uses were associated with increased breast cancer risk. Continuous combined preparations were associated with the highest risk. Further studies are needed to disentangle the effects of the regimen and the progestin component.
First evidence that the antimalarial drug artesunate inhibits invasion and in vivo metastasis in lung cancer by targeting essential extracellular proteases
Authors: Rasheed Suhail Ahmed Kabeer , Efferth Thomas , Asangani Irfan Ahmed , Allgayer Heike
Published: March 2010

Despite progress in treatment, progressive non–small cell lung cancer (NSCLC) still limits survival dramatically, and novel therapeutic compounds are needed. Initial investigations suggest that artesunate (ART), an antimalarial drug, has antiproliferative capacities. However, antiinvasive and antimetastatic properties of ART in cancer have never been explored. Therefore, this first study was performed to (i) investigate if ART is able to inhibit invasion and metastasis in NSCLC and (ii) to identify first molecular targets and mechanisms mediating this ability. ART significantly impaired matrigel invasion of 6 NSCLC cell lines and inhibited urokinase‐type plasminogen activator (u‐PA) activity, ‐protein and ‐mRNA expression. Furthermore, in a PCR‐metastasis array, ART inhibited the expression of several matrix metalloproteinases (MMPs), especially MMP‐2 and MMP‐7 mRNA/protein. In luciferase reporter assays, ART downregulated MMP‐2‐, MMP‐7‐ and u‐PA‐promoter/‐enhancer activity, in parallel to AP‐1‐ and NF‐kB‐transactivation. Si‐RNA knockdown of u‐PA, MMP‐2 and MMP‐7 abolished ART's ability to inhibit invasion, confirming their role as essential mediators. In vivo, ART significantly impaired primary tumor growth and metastasis in the chicken embryo metastasis (CAM) model. In conclusion, this is the first study to show that ART considerably suppresses invasion and metastasis in NSCLC, specifically targeting transcription of u‐PA, MMP‐2 and MMP‐7, prompting immediate studies on ART as a novel therapeutic in NSCLC.
Isolation and characterization of tumorigenic extrahepatic cholangiocarcinoma cells with stem cell‐like properties
Authors: Wang Min , Xiao Juan , Shen Min , Yahong Yu , Tian Rui , Zhu Feng , Jiang Jianxin , Du Zhiyong , Hu Jun , Liu Wensong , Qin Renyi
Published: March 2010

Recent studies suggest that the ability to form and grow tumors specifically resides in a small cell population called cancer stem cells (CSCs). These studies were conducted mainly on various human cancers; however, isolation and characterization of stem cells from cholangiocarcinoma have not been attempted. The molecular markers CD24, CD44, CD34, and epithelial cell adhesion molecule (EpCAM) are widely used, individually or in combination, to characterize some types of CSCs. In this study, we used these markers to identify a subpopulation of cells in extrahepatic cholangiocarcinoma (ECC) with cancer stem/progenitor cell‐like properties. We found that CD24+CD44+EpCAMhigh cells (0.39–2.27%) were present in human ECC tissues. The expression of a CD24+CD44+EpCAMhigh subpopulation was consistent with primary cancers and could be duplicated during serial in vivo passaging in NOD/SCID mice. CD24+CD44+EpCAMhigh cells isolated from 3 cholangiocarcinoma xenografts showed high tumorigenic potential compared with CD24−CD44−EpCAMlow/− cells. These tumorigenic ECC cells exhibited the stem cell properties of self‐renewal and ability to produce heterogeneous progeny. We report the identification of a CSC population in ECC characterized by CD24, CD44 and EpCAM phenotypes. Our findings could provide new insight into the tumorigenesis of cholangiocarcinoma and offer a potential target for anti‐cancer therapy.
Linoleic acid and butyrate synergize to increase Bcl‐2 levels in colonocytes
Authors: Turk Harmony F. , Kolar Satya S. , Fan Yang‐Yi , Cozby Caitlin A. , Lupton Joanne R. , Chapkin Robert S.
Published: March 2010

The biological properties of polyunsaturated fatty acid (PUFA) classes have been the source of much contention. For example, n‐3 PUFA are chemoprotective, whereas n‐6 PUFA may promote tumor development. Since dietary components can have combinatorial effects, we further examined the apoptotic properties of n‐3 or n‐6 fatty acids when combined with different fiber sources. Mice were fed diets supplemented with either fish oil (FO; enriched in n‐3 PUFA) or corn oil (CO; enriched in n‐6 PUFA) and nonfermentable (cellulose) or fermentable (pectin) fiber sources. In complementary experiments, immortalized young adult mouse colonic (YAMC) cells were treated with docosahexaenoic acid (DHA; 22:6n‐3) or linoleic acid (LA; 18:2n‐6) with or without butyrate. Mice fed a FO and pectin diet had significantly (p < 0.05) increased levels of apoptosis in colonocytes compared to all other diets. Similarly, apoptosis was highly induced in DHA and butyrate cotreated YAMC cells. In contrast, in both YAMC and mouse models, LA/CO with butyrate/pectin treatment reduced apoptosis and enhanced expression of bcl‐2. The LA and butyrate induced antiapoptotic phenotype was reversed by knocking down bcl‐2 using targeted siRNA. In comparison, overexpression of bcl‐2 blocked the proapoptotic effect of DHA and butyrate. These data provide new mechanistic insights into the regulation of apoptosis by dietary PUFA and fiber.
Gene expression changes induced by the human carcinogen aristolochic acid I in renal and hepatic tissue of mice
Authors: Arlt Volker M. , Zuo Jie , Trenz Kristina , Roufosse Candice A. , Lord Graham M. , Nortier Joelle L. , Schmeiser Heinz H. , Hollstein Monica , Phillips David H.
Published: March 2010

Aristolochic acid (AA) is the causative agent of urothelial tumors associated with AA nephropathy and is also implicated in the development of Balkan endemic nephropathy‐associated urothelial tumors. These tumors contain AA‐characteristic TP53 mutations. We examined gene expression changes in Hupki (human TP53 knock‐in) mice after treatment with aristolochic acid I (AAI) by gavage (5 mg/kg body weight). After 3, 12 and 21 days of treatment gene expression profiles were investigated using Agilent Whole Mouse 44K Genome Oligo Array. Expression profiles were significantly altered by AAI treatment in both target (kidney) and nontarget (liver) tissue. Renal pathology and DNA adduct analysis confirmed kidney as the target tissue of AAI‐induced toxicity. Gene ontology for functional analysis revealed that processes related to apoptosis, cell cycle, stress response, immune system, inflammatory response and kidney development were altered in kidney. Canonical pathway analysis indicated Nfκb, aryl hydrocarbon receptor, Tp53 and cell cycle signaling as the most important pathways modulated in kidney. Expression of Nfκb1 and other Nfκb‐target genes was confirmed by quantitative real‐time PCR (qRT‐PCR) and was consistent with the induction of Nfκb1 protein. Myc oncogene, frequently overexpressed in urothelial tumors, was upregulated by AAI on the microarrays and confirmed by qRT‐PCR and protein induction. Collectively we found that microarray gene expression analysis is a useful tool to define tissue‐specific responses in AAI‐induced toxicity. Several genes identified such as TP53, Rb1, Mdm2, Cdkn2a and Myc are frequently affected in human urothelial cancer, and may be valuable prognostic markers in future clinical studies.
No effect of meat, meat cooking preferences, meat mutagens or heme iron on lung cancer risk in the prostate, lung, colorectal and ovarian cancer screening trial
Authors: Tasevska Nataša , Cross Amanda J. , Dodd Kevin W. , Ziegler Regina G. , Caporaso Neil E. , Sinha Rashmi
Published: March 2010

Recent epidemiological studies have suggested that red and processed meat may increase the risk of lung cancer. Possible underlying mechanisms include mutagens produced during high‐temperature cooking or preservation, or formed endogenously from heme iron in meat. We used data from 99,579 participants of both screened and nonscreened arms of the Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial, aged 55–74 years, to investigate whether meat type, cooking method, doneness level, intake of specific meat mutagens 2‐amino‐3,8‐dimethylimidazo[4,5‐f]quinoxaline (MeIQx), 2‐amino‐3,4,8‐trimethylimidazo[4,5‐f]quinoxaline] (DiMeIQx), 2‐amino‐1‐methyl‐6‐phenylimidazo[4,5‐b]pyridine (PhIP) and benzo(a)pyrene (B(a)P)] and heme iron are associated with lung cancer. Participants' diet was assessed prospectively using a 124‐item food frequency questionnaire and an additional meat‐cooking module. Dietary data were used in conjunction with a database to estimate intake of MeIQx, DiMeIQx, PhIP, B(a)P and heme iron. After up to 8 years of follow‐up, 782 incident lung cancer cases were ascertained. Lung cancer risk was not associated with the consumption of either red (men: HRQ5vs.Q1= 1.11, 95% CI = 0.79–1.56,Ptrend= 0.42; women: HRQ5vs.Q1= 1.30, 95% CI = 0.87–1.95,Ptrend= 0.65) or processed meat (men: HRQ5vs.Q1= 1.12, 95% CI = 0.83–1.53,Ptrend= 0.22; women: HRQ5vs.Q1= 0.98, 95% CI = 0.68–1.41,Ptrend= 0.32) in multivariable models. High‐temperature cooking methods, level of meat doneness, meat mutagens and heme iron had no effect on lung cancer risk. In this population, we found no association between meat type, cooking method, doneness level or intake of specific meat mutagens or heme iron and lung cancer risk.
Transcriptional regulation of Rex1 (zfp42) in normal prostate epithelial cells and prostate cancer cells
Authors: Lee Mi‐Young , Lu Ailan , Gudas Lorraine J.
Published: March 2010

Rex1 (zfp42) was identified by our laboratory because of its reduced expression in F9 teratocarcinoma stem cells after retinoic acid (RA) treatment. The Rex1 (Zfp42) gene is currently widely used as a marker of embryonic stem cells. We compared the transcriptional regulation of the human Rex1 gene in NTera‐2 (NT‐2) human teratocarcinoma, normal human prostate epithelial cells (PrEC), and prostate cancer cells (PC‐3) by promoter/luciferase analyses. Oct4, Sox2, Nanog, and Dax1 transcripts are expressed at higher levels in NT‐2 and PrEC cells than in PC‐3 cells. Co‐transfection analyses showed that YY1 and Rex1 are positive regulators of hRex1 transcription in NT‐2 and PrEC cells, whereas Nanog is not. Serial deletion constructs of the hRex1 promoter were created and analyzed, by which we identified a potential negative regulatory site that is located between −1 and −0.4 kb of the hRex1 promoter. We also delineated regions of the hRex1 promoter between −0.4 kb and the TSS that, when mutated, reduced transcriptional activation; these are putative Rex1 binding sites. Mutation of a putative Rex1 binding site in electrophoretic mobility shift assays (EMSA) resulted in reduced protein binding. Taken together, our results indicate that hRex1 binds to the hRex1 promoter region at −298 bp and positively regulates hRex1 transcription, but that this regulation is lost in PC‐3 human prostate cancer cells. This lack of positive transcriptional regulation by the hRex1 protein may be responsible for the lack of Rex1 expression in PC‐3 prostate cancer cells. J. Cell. Physiol. 224:17–27, 2010 © 2010 Wiley‐Liss, Inc.
Stromal cell‐derived factor 1α reduces senescence of endothelial progenitor subpopulation in lectin‐binding and DiLDL‐uptaking cell through telomerase activation and telomere elongation
Authors: Zheng Hao , Shen Cheng‐Ji , Qiu Fu‐Yu , Zhao Yan‐Bo , Fu Guo‐Sheng
Published: March 2010

Recent studies have suggested that reduced endothelial progenitor subpopulation in lectin‐binding and DiLDL‐uptaking cell (EPC subpopulation) number and activity was associated with EPC subpopulation senescence that involved telomerase activity and telomere length. Stromal cell‐derived factor‐1α (SDF‐1α) has been shown to augment a variety of cellular functions of EPC subpopulation and subsequently contribute to ischemic neovascularization. Therefore, we investigated whether SDF‐1α might be able to prevent senescence of EPC subpopulation and also investigated the effects of SDF‐1α on the telomerase activity and telomere length. EPC subpopulation were isolated from peripheral blood and characterized. After ex vivo prolonged cultivation, EPC subpopulation became senescent as determined by acidic β‐galactosidase staining. SDF‐1α dose‐dependently inhibited the onset of EPC subpopulation senescence. Moreover, SDF‐1α increased proliferation and colony‐forming activity of EPC subpopulation. SDF‐1α also increased telomerase activity and telomere length, which was accompanied with upregulation of the catalytic subunit, telomerase reverse transcriptase (TERT). Whereas these effects of SDF‐1α on telomerase activity and expression of hTERT mRNA were significantly attenuated by CXCR4‐specific peptide antagonist (AMD3100) and phosphoinositide 3‐kinase (PI3K) inhibitor (LY294002). In conclusions, SDF‐1α delays the onset of EPC subpopulation senescence, which may be related to the activation of telomerase and elongation of telomere length. The inhibition of EPC subpopulation senescence and induction of EPC subpopulation proliferation by SDF‐1α in vitro may importantly improve the functional activity of EPC subpopulation for potential cell therapy. J. Cell. Physiol. 223:757–763, 2010. © 2010 Wiley‐Liss, Inc.
Characterization of BAG3 cleavage during apoptosis of pancreatic cancer cells
Authors: Wang Hua‐Qin , Meng Xin , Gao Yan‐Yan , Liu Bao‐Qin , Niu Xiao‐Fang , Zhang Hai‐Yan , Du Zhen‐Xian
Published: March 2010

Caspases are a conserved family of cell death proteases that cleave intracellular substrates at Asp residues to modify their function and promote apoptosis. In this report, we identify BAG3 as a novel caspases substrate. Here, we show that one of these BAG proteins, BAG3, is cleaved during apoptosis. BAG3 cleavage is inhibited by several different caspase inhibitors. The analysis of BAG3 cleavage by recombinant caspase proteins shows that BAG3 is efficiently cleaved by caspase‐3, to a smaller extent by caspases‐1 and ‐8, and relatively inefficient by caspase‐9. Cleavage of the BAG3 protein occurs in the C‐terminal part of the protein majorly at Asp347 (KEVD347↓S) in vitro and in pancreatic cancer SW1990 and PANC‐1 cells undergoing apoptosis. We also demonstrate that unlike cleavage of Bcl‐2 and Bcl‐XL, cleaved form of BAG3 does not result in pro‐apoptotic fragments, however, cleavage of BAG3 lead to loss its per se anti‐apoptotic property. This novel regulation of BAG3 may have important implications for its role in apoptosis. J. Cell. Physiol. 224:94–100, 2010 © 2010 Wiley‐Liss, Inc.
Growth differentiation factor 15 acts anti‐apoptotic and pro‐hypertrophic in adult cardiomyocytes
Authors: Heger J. , Schiegnitz E. , von Waldthausen D. , Anwar M.M. , Piper H.M. , Euler G.
Published: March 2010

Growth differentiation factor 15 (GDF15) is induced during heart failure development, and may influence different processes in cardiac remodeling. While its anti‐apoptotic action under conditions of ischemia–reperfusion have been shown, it remained unclear if this is a broadly protective effect applicable to other apoptotic stimuli. Furthermore, effects on cardiac hypertrophy remained obscure. Therefore, we investigated the effects of GDF15 on induction of hypertrophy and apoptosis in ventricular cardiomyocytes. GDF15 (3 ng/ml) enhanced hypertrophic growth of cardiomyocytes as determined by an increase in cell size by 27 ± 5% and rate of protein synthesis by 47 ± 15%. In addition, a time and dose‐dependent increase in SMAD‐binding affinity was found, as well as enhanced phosphorylation of R‐SMAD1. Inhibition of SMADs by transformation of cardiomyocytes with SMAD‐decoy oligonucleotides abolished the hypertrophic growth effect. Specific inhibitors of PI3K (10 µM LY290042 or 10 nM wortmannin) or ERK (10 µM PD98059) also blocked GDF15‐induced hypertrophy and SMAD activation. Apoptosis induction by three different agents, 100 nM angiotensin II, 1 ng/ml TGFβ1, or the NO‐donor SNAP (100 µM) was blocked by addition of GDF15 (3 ng/ml). Scavenging of SMADs by transformation of cardiomyocytes with SMAD‐decoy oligonucleotides abolished the anti‐apoptotic effect of GDF15. In conclusion, GDF15 protects ventricular cardiomyocytes against different apoptotic stimuli and enhances hypertrophic growth. Hypertrophic signaling is thereby mediated via the kinases PI3K and ERK and the transcription factor R‐SMAD1. Thus, GDF15 may influence cardiac remodeling via two different mechanisms, apoptosis protection and induction of hypertrophy. J. Cell. Physiol. 224:120–126, 2010 © 2010 Wiley‐Liss, Inc.
Group V secreted phospholipase A2contributes to LPS‐induced leukocyte recruitment
Authors: Lapointe Stéphanie , Brkovic Alexandre , Cloutier Isabelle , Tanguay Jean‐François , Arm Jonathan P. , Sirois Martin G.
Published: March 2010

Secreted phospholipases A2(sPLA2s) are well known for their contribution in the biosynthesis of inflammatory eicosanoids. These enzymes also participate in the inflammatory process by regulating chemokine production and protein expression of adhesion molecules. The majority of sPLA2isoforms are up‐regulated by proinflammatory stimuli such as bacterial lipopolysaccharide (LPS), which predominantly increases the expression of group V sPLA2(sPLA2‐V). Furthermore, it has recently been shown that sPLA2‐V is a critical messenger in the regulation of cell migration during allergic airway responsiveness. Herein, we investigated the effect of sPLA2‐V on LPS‐mediated leukocyte recruitment and its capacity to modulate adhesion molecule expression. We conducted our study in the murine air pouch model, using sPLA2‐V null mice (sPLA2‐V−/−) and control wild‐type (WT) littermates. We observed that LPS (1 µg/ml)‐mediated leukocyte emigration in sPLA2‐V−/−was attenuated by 52% and 86% upon 6 and 12 h of treatment respectively, as compared to WT mice. In WT mice, treatment with the cell‐permeable sPLA2inhibitor (12‐epi‐scalaradial; SLD) reduced LPS‐mediated leukocyte recruitment by 67%, but had no additional inhibitory effect in sPLA2‐V−/−mice. Protein analyses from the air pouch skin were carried out upon LPS‐challenge, and the expression of intercellular adhesion molecule (ICAM)‐1 and vascular cell adhesion molecule (VCAM)‐1 were both significantly reduced in sPLA2‐V−/−mice as compared to control WT mice. Together, our data demonstrate the role of sPLA2‐V in LPS‐induced ICAM‐1 and VCAM‐1 protein overexpression and leukocyte recruitment, supporting the contribution of sPLA2‐V in the development of inflammatory innate immune responses. J. Cell. Physiol. 224:127–134, 2010 © 2010 Wiley‐Liss, Inc.
TGFβ mediated transition of corneal fibroblasts from a proinflammatory state to a profibrotic state through modulation of histone acetylation
Authors: Zhou Qingjun , Yang Lingling , Wang Yao , Qu Mingli , Chen Peng , Wang Ye , Xie Lixin , Zhao Jing , Wang Yiqiang
Published: March 2010

Corneal fibroblasts exhibit different phenotypes in different phases of corneal wound healing. In the inflammatory phase, the cells assume a proinflammatory phenotype and produce large amounts of cytokines and chemokines, but in the proliferative and remodeling phases, they adapt a profibrotic state, differentiate into myofibroblasts and increase extracellular matrix protein synthesis, secretion, and deposition. In the present study, the molecular mechanisms regulating the transition of corneal fibroblasts from the proinflammatory state to the profibrotic state were investigated. Corneal fibroblasts were treated with TGFβ, a known profibrotic and anti‐inflammatory factor in wound healing, in the absence or presence of trichostatin A (TSA), a histone deacetylase (HDAC) inhibitor. The results revealed that TGFβ induced the profibrotic transition of corneal fibroblasts, including increased extracellular matrix synthesis, morphological changes, and assembly of actin filaments. Meanwhile, proinflammatory gene expressions of corneal fibroblasts were down‐regulated with the treatment of TGFβ, as confirmed by cDNA microarray, real time PCR and ELISA. Moreover, TSA reversed the TGFβ‐mediated transition of corneal fibroblasts from the proinflammatory state to the profibrotic state, as accompanied by histone hyperacetylations. In conclusion, TGFβ suppressed the production of proinflammatory factors and enhanced the expression of matrix remodeling genes of corneal fibroblasts in the transition from the proinflammatory state to the profibrotic state, and the dual roles of TGFβ on the phenotype regulations of corneal fibroblasts were mediated by altered histone acetylation. J. Cell. Physiol. 224:135–143, 2010 © 2010 Wiley‐Liss, Inc.
miR‐20b modulates VEGF expression by targeting HIF‐1α and STAT3 in MCF‐7 breast cancer cells
Authors: Cascio Sandra , D'Andrea Aleco , Ferla Rita , Surmacz Eva , Gulotta Eliana , Amodeo Valeria , Bazan Viviana , Gebbia Nicola , Russo Antonio
Published: March 2010

MicroRNAs (miRNAs) are small non‐coding RNAs that regulate the expression of different genes, including genes involved in cancer progression. A functional link between hypoxia, a key feature of the tumor microenvironment, and miRNA expression has been documented. We investigated whether and how miR‐20b can regulate the expression of vascular endothelial growth factor (VEGF) in MCF‐7 breast cancer cells under normoxic and hypoxia‐mimicking conditions (CoCl2 exposure). Using immunoblotting, ELISA, and quantitative real‐time PCR, we demonstrated that miR‐20b decreased VEGF protein levels at 4 and 24 h following CoCl2 treatment, and VEGF mRNA at 4 h of treatment. In addition, miR‐20b reduced VEGF protein expression in untreated cells. Next, we investigated the molecular mechanism by which pre‐miR‐20b can affect VEGF transcription, focusing on hypoxia inducible factor 1 (HIF‐1) and signal transducer and activator of transcription 3 (STAT3), transcriptional inducers of VEGF and putative targets of miR‐20b. Downregulation of VEGF mRNA by miR‐20b under a 4 h of CoCl2 treatment was associated with reduced levels of nuclear HIF‐1α subunit and STAT3. Chromatin immunoprecipitation (ChIP) assays revealed that HIF‐1α, but not STAT3, was recruited to the VEGF promoter following the 4 h of CoCl2 treatment. This effect was inhibited by transfection of cells with pre‐miR‐20b. In addition, using siRNA knockdown, we demonstrated that the presence of STAT3 is necessary for CoCl2‐mediated HIF‐1α nuclear accumulation and recruitment on VEGF promoter. In summary, this report demonstrates, for the first time, that the VEGF expression in breast cancer cells is mediated by HIF‐1 and STAT3 in a miR‐20b‐dependent manner. J. Cell. Physiol. 224:242–249, 2010 © 2010 Wiley‐Liss, Inc.
3D reconstruction of histological sections: Application to mammary gland tissue
Authors: Arganda‐Carreras Ignacio , Fernández‐González Rodrigo , Muñoz‐Barrutia Arrate , Ortiz‐De‐Solorzano Carlos
Published: March 2010

In this article, we present a novel method for the automatic 3D reconstruction of thick tissue blocks from 2D histological sections. The algorithm completes a high‐content (multiscale, multifeature) imaging system for simultaneous morphological and molecular analysis of thick tissue samples. This computer‐based system integrates image acquisition, annotation, registration, and three‐dimensional reconstruction. We present an experimental validation of this tool using both synthetic and real data. In particular, we present the 3D reconstruction of an entire mouse mammary gland and demonstrate the integration of high‐resolution molecular data. Microsc. Res. Tech. 73:1019–1029, 2010. © 2010 Wiley‐Liss, Inc.
Structural and functional consequences of poly(ethylene glycol) inclusion on DNA condensation for gene delivery
Authors: Millili Peter G. , Selekman Joshua A. , Blocker Kory M. , Johnson David A. , Naik Ulhas P. , Sullivan Millicent O.
Published: March 2010

Polycationic polymers have been used to condense therapeutic DNA into submicron particles, offering protection from shear‐induced or enzymatic degradation. However, the spontaneous nature of this self‐assembly process gives rise to the formation of multimolecular aggregates, resulting in significant polyplex heterogeneity. Additionally, cytotoxicity issues and serum instability have limited the in vivo efficacy of such systems. One way these issues can be addressed is through the inclusion of poly(ethylene glycol) (PEG). PEG has known steric effects that inhibit polyplex self‐aggregation. A variety of PEGylated gene delivery formulations have been previously pursued in an effort to take advantage of this material's benefits. Because of such interest, our aim was to further explore the consequences of PEG inclusion on the structure and activity of gene delivery vehicle formulations. We explored the complexation of plasmid DNA with varying ratios of a PEGylated trilysine peptide (PEG‐K3) and 25‐kDa polyethylenimine (PEI). Atomic force and scanning electron microscopy were utilized to assess the polyplex size and shape and revealed that a critical threshold of PEG was necessary to promote the formation of homogeneous polyplexes. Flow cytometry and fluorescence microscopy analyses suggested that the presence of PEG inhibited transfection efficiency as a consequence of changes in intracellular trafficking and promoted an increased reliance on energy‐independent mechanisms of cellular uptake. These studies provide new information on the role of PEG in delivery vehicle design and lay the foundation for future work aimed at elucidating the details of the intracellular transport of PEGylated polyplexes. Microsc. Res. Tech. 73:866–877, 2010. © 2010 Wiley‐Liss, Inc.
Two and three‐dimensional gene transfer from enzymatically degradable hydrogel scaffolds
Authors: Lei Yuguo , Ng Quinn K.T. , Segura Tatiana
Published: March 2010

The ability to genetically modify mesenchymal stem cells (MSCs) seeded inside synthetic hydrogel scaffolds would offer an alternative approach to guide MSC differentiation. In this report, we explored gene transfer to MSCs seeded on top or inside matrix metalloproteinase (MMP) degradable hydrogels that were loaded with DNA/poly(ethylene imine) (PEI) polyplexes. DNA/PEI polyplexes were encapsulated inside poly(ethylene glycol) (PEG) hydrogels crosslinked with MMP degradable peptides via Michael Addition chemistry. Gene transfer was visualized and quantified through using a vector encoding for green fluorescent protein and luciferase. We found that gene transfer to MSCs was possible for cells seeded both in two and three dimensions. The amount of luciferase expression was similar for cells seeded in two and three dimensions even though the number of cells in three dimensions is significantly higher, indicating that gene transfer to cells seeded in two dimensions is more efficient than for cells seeded in three dimensions. The use of hydrogel scaffolds that allow cellular infiltration to deliver DNA may result in long‐lasting signals in vivo, which are essential for the regeneration of functional tissues. Microsc. Res. Tech. 73:910–917, 2010. © 2010 Wiley‐Liss, Inc.