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Background The obesity pandemic is associated with multiple major health concerns. In addition to diet and lifestyle, there is increasing evidence that environmental exposures to chemicals known as obesogens also may promote obesity. Objectives We investigated the massive environmental contamination resulting from the Deepwater Horizon (DWH) oil spill, including the use of the oil dispersant COREXIT in remediation efforts, to determine whether obesogens were released into the environment during this incident. We also sought to improve the sensitivity of obesogen detection methods in order to guide post-toxicological chemical assessments. Methods Peroxisome proliferator–activated receptor gamma (PPARγ) transactivation assays were used to identify putative obesogens. Solid-phase extraction (SPE) was used to sub-fractionate the water-accommodated fraction generated by mixing COREXIT, cell culture media, and DWH oil (CWAF). Liquid chromatography–mass spectrometry (LC-MS) was used to identify components of fractionated CWAF. PPAR response element (PPRE) activity was measured in PPRE-luciferase transgenic mice. Ligand-binding assays were used to quantitate ligand affinity. Murine 3T3-L1 preadipocytes were used to assess adipogenic induction. Results Serum-free conditions greatly enhanced the sensitivity of PPARγ transactivation assays. CWAF and COREXIT had significant dose-dependent PPARγ transactivation activities. From SPE, the 50:50 water:ethanol volume fraction of CWAF contained this activity, and LC-MS indicated that major components of COREXIT contribute to PPARγ transactivation in the CWAF. Molecular modeling predicted several components of COREXIT might be PPARγ ligands. We classified dioctyl sodium sulfosuccinate (DOSS), a major component of COREXIT, as a probable obesogen by PPARγ transactivation assays, PPAR-driven luciferase induction in vivo, PPARγ binding assays (affinity comparable to pioglitazone and arachidonic acid), and in vitro murine adipocyte differentiation. Conclusions We conclude that DOSS is a putative obesogen worthy of further study, including epidemiological and clinical investigations into laxative prescriptions consisting of DOSS. Citation Temkin AM, Bowers RR, Magaletta ME, Holshouser S, Maggi A, Ciana P, Guillette LJ, Bowden JA, Kucklick JR, Baatz JE, Spyropoulos DD. 2016. Effects of crude oil/dispersant mixture and dispersant components on PPARγ activity in vitro and in vivo: identification of dioctyl sodium sulfosuccinate (DOSS; CAS #577-11-7) as a probable obesogen. Environ Health Perspect 124:112–119; http://dx.doi.org/10.1289/ehp.1409672

Estrogen deprivation is associated with delayed healing, while estrogen replacement therapy (ERT) accelerates acute wound healing and protects against development of chronic wounds. However, current estrogenic molecules have undesired systemic effects, thus the aim of our studies is to generate new molecules for topic administration that are devoid of systemic effects. Following a preliminary study, the new 17β-estradiol derivatives 1 were synthesized. The estrogenic activity of these novel compounds was evaluated in vitro using the cell line ERE-Luc B17 stably transfected with an ERE-Luc reporter. Among the 17β-estradiol derivatives synthesized, compounds 1e and 1f showed the highest transactivation potency and were therefore selected for the study of their systemic estrogenic activity. The study of these compounds in the ERE-Luc mouse model demonstrated that both compounds lack systemic effects when administered in the wound area. Furthermore, wound-healing experiments showed that 1e displays a significant regenerative and anti-inflammatory activity. It is therefore confirmed that this class of compounds are suitable for topical administration and have a clear beneficial effect on wound healing.

In vitro polarization responses of microglia to IL-4 and CSF-1. A) The expression of M2 polarization genes (Arg1 and Mrc1) was analyzed in primary cultures of microglia following the treatment with vehicle (veh; open boxes), 20 ng/ml IL-4 (closed boxes) or 20 ng/ml CSF-1 (gray boxes). B) The expression of IL-4Rα was analyzed either in microglia cells obtained by immunosorting from the brain of vehicle or IL-4 icv-injected mice (ex vivo) or in primary cultures of microglia (in vitro) as specified above. Bars represent the mean ± SEM of 3 independent experiments, each performed in triplicate. Data sets for each gene were calculated using the 2-ddCt method with respect to the mean value of the vehicle group. Bars represent mean values ± SEM (n = 3). Student’s unpaired t-test, *p

In vitro proliferative and polarization responses of macrophages to IL-4, CSF-1 and E2. A) The expression of proliferation (Ki67, Cdk1 and Ccnb2) and M2 polarization (Arg1, Fizz1, Mrc1) genes was analyzed in primary cultures of peritoneal macrophages following treatment with 20 ng/ml IL-4 (black bars) or 20 ng/ml CSF-1 (gray bars). B) The expression of proliferation (Ki67, Cdk1 and Ccnb2) and M2 polarization (Arg1, Vegfα) genes was analyzed in primary cultures of peritoneal macrophages following vehicle (0.01% EtOH) or 10−5 M E2 treatments. Bars represent the mean ± SEM of 3 independent experiments, each performed in triplicate Data sets for each gene were calculated using the 2-ddCt method with respect to the mean value of the vehicle group. Bars represent mean values ± SEM (n = 3). Student’s unpaired t-test, *p

Cellular localization of Ki67 immunostaining in brain astrocytes. Brain sections were analyzed by immunohistochemistry for the expression of Ki67 using antibodies against Ki67 (green labeling) and GFAP (red staining) following the icv administration of 250Â ng IL-4. Ki67-positive cells co-localize with GFAP-positive cells (white arrows). Scale bar, 10Â Îźm. (PDF 105 kb)

Deciphering the etiology of complex pathologies at molecular level requires longitudinal studies encompassing multiple biochemical pathways (apoptosis, proliferation, inflammation, oxidative stress). In vivo imaging of current reporter animals enabled the spatio-temporal analysis of specific molecular events, however, the lack of a multiplicity of loci for the generalized and regulated expression of the integrated transgenes hampers the creation of systems for the simultaneous analysis of more than a biochemical pathways at the time. We here developed and tested an in vivo-based methodology for the identification of multiple insertional loci suitable for the generation of reliable reporter mice. The validity of the methodology was tested with the generation of novel mice useful to report on inflammation and oxidative stress.

Some fatty acids, such as CLA (conjugated linoleic acid) and n-3 fatty acids modulate immune and inflammatory response in ruminants and monogastrics; their supplementation alters fatty acids profile of meat and milk, enhancing their nutritional quality. However, it is still unclear if their addition causes oxidative damage to animals. Nuclear factor E2-related factor 2 (Nrf2) plays an important role in cellular defenses against oxidative stress, indeed it produces a rapid induction of its target genes involved in antioxidant response. The aim of the project is to investigate the activation of Nrf2 in luciferase reporter mice fed different amount of n-3 PUFA in the diet (7,5% lard, 7,5% tuna oil, 20 % lard and 20%tuna oil). Forty-eight reporter mice are divided into three groups: male, intact female and ovariectomized female. Each group is split in four subgroups fed different diets. Oxidative status will be studied monitoring Nrf2’s activation with in vivo bioluminescent imaging. The inflammatory and immune response will be assessed using calprotectin and lactoferrin levels in faecal samples that are non-invasive techniques. The trial is still in progress: on the 62nd day, animals will be sacrificed after a challenge in order to measure the different effects of diets and +/- oestrogen on stress response. Finally, the post mortem analysis will be carried on extract organs. Data obtained will be analysed using statistical procedures and results will improve the knowledge about interaction between omega-3 fatty acids and animals’ oxidative status.

Inflammatory activation of microglia is a hallmark of several disorders of the CNS. In addition to protecting the brain against inflammatory insults, microglia are neuroprotective and play a significant role in maintaining neuronal connectivity; therefore, the prolongation and inflammatory status may limit the beneficial functions of these immune cells. The findings that estrogen receptors are present in monocyte-derived cells and that estrogens prevent and control the inflammatory response raise the question of the role that this sex hormone plays in the manifestation and progression of pathologies that have a clear sex difference in prevalence, such as multiple sclerosis, Parkinson's disease, and Alzheimer's disease. The present review aims to provide a critical review of the current literature on the actions of estrogen in microglia and on the involvement of estrogen receptors in the manifestation of selected neurological disorders. This current understanding highlights a research area that should be expanded to identify appropriate replacement therapies to slow the progression of such diseases.

In female animals, energy metabolism and fertility are tightly connected, and reciprocally regulated. However, the relative contributions of metabolic and reproductive pathways have changed over the course of evolution. In oviparous animals, metabolic factors take precedence over fertility, enabling egg production to be inhibited in a nutritionally poor environment. By contrast, in placental mammals, the opposite occurs: the need to feed a developing embryo and neonate forces metabolic pathways to adapt to these reproductive needs. This physiological necessity explains why in female mammals alterations of gonadal activity, including age-dependent cessation of ovarian functions, are associated with a disruption of metabolic homeostasis and consequent inflammatory reactions that trigger the onset of metabolic, cardiovascular, skeletal and neural pathologies. This Review discusses how metabolic homeostasis and reproductive functions interact to optimize female fertility and explains the pathogenic mechanisms underlying the disordered energy metabolism associated with human ovarian dysfunction owing to menopause, polycystic ovary syndrome and Turner syndrome. Finally, this article highlights how hormone replacement therapy might aid the restoration of metabolic homeostasis in women with ovarian dysfunction.

La intervención sobre el tejido urbano antiguo de Ciutat Vella es una de las obras menos conocidas del estudio de Miralles recogida en el PERI BA 189-2 de 1997, pero de un gran valor. Analizaré la intervención urbana de Miralles en relación a intervenciones anteriores, para mostrar la importancia del cambio que se produjo en la manera de intervenir en el centro histórico de Barcelona.