Determinants of increasing serum POPs in a population at high risk for cardiovascular disease. Results from the PREDIMED-CANARIAS study.

Persistent organic pollutants (POPs) are well-known ubiquitous environmental chemicals which have been related to adverse health outcomes, including cardiovascular disease (CVD). The purpose of this study was to evaluate POPs burden, and its determinants, in a population at high risk of suffering CVD enrolled in the PREDIMED Study (Spanish acronym for PREvention by means of MEDiterranean Diet). This cohort was formed by 343 participants (55-80 y.o.), which were selected for a preventive nutritional intervention for CVD based on the Mediterranean Diet. Relevant information on demographic, behavioral, dietary, and socioeconomic characteristics was obtained from each participant through a specific questionnaire, and their anthropometric and clinical measurements were recorded. In addition, the levels of 35 POPs were determined in serum samples taken before the beginning of the nutritional intervention. All the samples showed detectable levels of, at least, one POP, being DDT-derivatives and marker-PCBs the most frequently detected compounds. Our results showed that people at high risk for CVD showed a higher level of contamination by POPs as compared to other studies done in cohorts of Western people at no special risk of CVD. Although educational level seems to be a relevant determinant for POPs burden in our population, the main determining factor seems to be the diet. Thus, while the intake of food of animal origin was significantly associated with levels of PCBs, especially in men, the intake of vegetal-origin food was positively related to levels of organochlorine pesticides, indicating a different dietary source for these two groups of chemicals. Our results showing that subjects at high risk for cardiovascular disease present elevated POPs burden might have a relevant public health impact given the generalized and difficult to avoid exposure to POPs and the elevated worldwide frequency of the CVD.

Relationship of polychlorinated biphenyls (PCBs) with parasitism, iron homeostasis, and other health outcomes: Results from a cross-sectional study on recently arrived African immigrants.

Polychlorinated biphenyls (PCBs) are toxic and persistent chemicals produced between 1930s and 1980s, which accumulate in humans and wildlife. Although a decreasing trend of PCB levels in humans has been described in developed countries, mainly as a consequence of strict regulations and remediation plans, an inverse trend has been recently reported in people from developing countries. We had the opportunity of sampling a series of African immigrants recently arrived to the Spanish archipelago of the Canary Islands, in which high levels of PCBs have been described, and we studied the relationships between their level of contamination and health status. A total of 570 subjects who underwent a complete medical examination and a face-to-face interview were recruited for this study. Hematological and biochemical parameters (blood and urine) were determined in all participants. Serology for the diagnostic of infectious diseases was also performed, and direct identification of parasites was performed in feces, urine or blood samples when appropriate. It is remarkable that up to 26.0% of the population had intestinal parasites, and we found an inverse relationship between PCB levels and parasitism and parasitic diseases: median values of PCBs were lower in parasitized subjects than in subjects without parasites in stool (237.6ng/g fat vs. 154.4ng/g fat for marker PCBs, p=0.015) and median values of dioxin-like PCBs were lower in subjects carrying pathogen parasites than among subjects showing non-pathogen parasites in stool (0.0 ng/g fat vs. 13.1ng/g fat, respectively; p=0.001). Although this inverse association had been described in some vertebrates this is the first study reporting such an association in humans. Furthermore, it has been also recently described that PCBs may disrupt iron metabolism, and we found a direct relationship between serum iron and total PCBs burden (r=0.231, p=0.025), suggesting that PCBs, although at subclinical level, could play a role on iron homeostasis. Although the role of PCBs in parasitism and in the iron metabolism needs future research, our findings may help to understand the adverse health outcomes associated to environmental exposure to PCBs and they might be used in exposed populations as indicators of subtle effects due to environmental insult.

Results of Total Maxillary Edentulism Rehabilitation Protocol with Implant-Supported Prostheses in the Pterygomaxillary-Pyramidal Region as Posterior Anchorage Loci.

PURPOSE: To present functional and esthetic success of a well-defined protocol for rehabilitation with implant-supported complete-arch prostheses in the treatment of patients with maxillary edentulism. MATERIALS AND METHODS: Medical records of patients treated from 1990 to September 2014 were reviewed. This clinic history should include clinical, anatomicalfunctional, and esthetic data for at least the first 5 years of follow-up. RESULTS: Two hundred eighty-two implants were inserted into the pterygomaxillary-pyramidal region in 141 edentulous patients using cylindrical osteotomes to form bone sites and thereby minimize the use of drills in bone preparation. Also, 580 implants were inserted in the premaxilla using lyophilized bone of bovine origin as a graft material and platelet-rich plasma for repairing small bone defects around implants and as a volume-generating material to support perioral structures. Of the 282 implants placed in the pterygomaxillary-pyramidal region, 272 survived (96.1% survival rate). The 10 failures occurred after being subjected to functional load. Of the 580 implants placed in the premaxilla, 19 implants failed; thus, the survival rate was 96.7%. Moreover, from the results obtained in the study sample, it was observed that the retention and stability obtained with the implant-supported prosthesis helps improve the ability to speak, swallow, and chew, which improves patient comfort and satisfaction. CONCLUSION: The rehabilitation of total maxillary edentulism with an implant-supported complete-arch prosthesis achieves a high level of functional and esthetic success, which is reflected directly in oral health and patient satisfaction.

Long-term results in placement of screw-type implants in the pterygomaxillary-pyramidal region.

PURPOSE: To present the long-term results for screw-type implants consecutively placed in the pterygomaxillary-pyramidal region in the treatment of patients with posterior maxillary edentulism. MATERIALS AND METHODS: One hundred fifty-two implants were inserted in 92 partially edentulous patients using cylindric osteotomes as bone-site formers, thus minimizing the use of drills in the bone preparation. RESULTS: The 152 implants placed were loaded for an average of 89.7+/-30.7 months. There were a total of 8 failures; 6 took place between implant placement and prosthesis delivery (early failures), and 2 failed in following functional loading. The overall survival rate was 94.7%. DISCUSSION: Cylindric osteotomes allow the surgeon to accurately place implants. The use of surgical drilling was minimized, resulting in lowered surgical risk and diminished bone loss. CONCLUSION: Implant placement in the posterior pterygomaxillary region using cylindric osteotomes for osteotomy preparation resulted in an implant survival rate of 94.7%.

ROCK and nuclear factor-kappaB-dependent activation of cyclooxygenase-2 by Rho GTPases: effects on tumor growth and therapeutic consequences.

Rho GTPases are overexpressed in a variety of human tumors contributing to both tumor proliferation and metastasis. Recently, several studies demonstrate an essential role of transcriptional regulation in Rho GTPases-induced oncogenesis. Herein, we demonstrate that RhoA, Rac1, and Cdc42 promote the expression of cyclooxygenase-2 (COX-2) at the transcriptional level by a mechanism that is dependent on the transcription factor nuclear factor-kappaB (NF-kappaB), but not Stat3, a transcription factor required for RhoA-induced tumorigenesis. With respect to RhoA, this effect is dependent on ROCK, but not PKN. Treatment of RhoA-, Rac1-, and Cdc42-transformed epithelial cells with Sulindac and NS-398, two well-characterized nonsteroid antiinflammatory drugs (NSAIDs), results in growth inhibition as determined by cell proliferation assays. Accordingly, tumor growth of RhoA-expressing epithelial cells in syngeneic mice is strongly inhibited by NS-398 treatment. The effect of NSAIDs over RhoA-induced tumor growth is not exclusively dependent on COX-2 because DNA-binding of NF-kappaB is also abolished upon NSAIDs treatment, resulting in complete loss of COX-2 expression. Finally, treatment of RhoA-transformed cells with Bay11-7083, a specific NF-kappaB inhibitor, leads to inhibition of cell proliferation. We suggest that treatment of human tumors that overexpress Rho GTPases with NSAIDs and drugs that target NF-kappaB could constitute a valid antitumoral strategy.

STAT5a activation mediates the epithelial to mesenchymal transition induced by oncogenic RhoA.

The involvement of Rho GTPases in signal transduction pathways leading to transcription activation is one of the major roles of this family of GTPases. Thus, the identification of transcription factors regulated by Rho GTPases and the understanding of the mechanisms of their activation and its biological outcome are of great interest. Here, we provide evidence that Rho GTPases modulate Stat5a, a transcription factor of the family of signal transducers and activators of transcription. RhoA triggers tyrosine phosphorylation (Y696) of Stat5a via a JAK2-dependent mechanism and promotes DNA-binding activity of Stat5a. Tyrosine phosphorylation of Stat5a is also stimulated physiologically by lysophosphatidic acid (LPA) in a Rho-dependent manner. Simultaneously, RhoA reduces serine phosphorylation of Stat5a at both serine residues S726 and S780, resulting in a further increase of activity as defined by mutagenesis experiments. Furthermore, serine dephosphorylation of Stat5a by RhoA does not take place by down-modulation of either JNK1, MEK1, or p38 MAP kinases, as determined by transfection experiments or chemical inhibition of both MEK1, p38, and JNK serine kinases. Thus, RhoA regulates Stat5a via tyrosine phosphorylation and via a yet to be determined novel down-modulating pathway that involves serine dephosphorylation. Finally, we provide evidence for a role of Stat5a in RhoA-induced epithelial-to-mesenchymal transition with concomitant increase in vimentin expression, E-cadherin down-regulation, and cell motility.

Consumption of organic meat does not diminish the carcinogenic potential associated with the intake of persistent organic pollutants (POPs).

Numerous studies have shown an epidemiological link between meat consumption and the incidence of cancer, and it has been suggested that this relationship may be motivated by the presence of carcinogenic contaminants on it. Among the most frequently detected contaminants in meat are several types of persistent organic pollutants (POPs), and it is well known that many of them are carcinogenic. On the other hand, an increasing number of consumers choose to feed on what are perceived as healthier foods. Thus, the number of consumers of organic food is growing. However, environmental contamination by POPs is ubiquitous, and it is therefore unlikely that the practices of organic food production are able to prevent this contamination. To test this hypothesis, we acquired 76 samples of meat (beef, chicken, and lamb) of two modes of production (organic and conventional) and quantified their levels of 33 carcinogenic POPs. On this basis, we determined the human meat-related daily dietary exposure to these carcinogens using as a model a population with a high consumption of meat, such as the Spanish population. The maximum allowable meat consumption for this population and the carcinogenic risk quotients associated with the current pattern of consumption were calculated. As expected, no sample was completely free of carcinogenic contaminants, and the differences between organically and conventionally produced meats were minimal. According to these results, the current pattern of meat consumption exceeded the maximum limits, which are set according to the levels of contaminations, and this is associated with a relevant carcinogenic risk. Strikingly, the consumption of organically produced meat does not diminish this carcinogenic risk, but on the contrary, it seems to be even higher, especially that associated with lamb consumption.

Persistent organic pollutants and risk of diabetes and obesity on healthy adults: Results from a cross-sectional study in Spain.

Environmental exposure to persistent organic pollutants (POPs) has been reported to be relevant in the population of the Canary Islands (Spain), especially that of organochlorine pesticides. On the other hand, the population of this archipelago presents a high prevalence of type 2 diabetes (T2D), and it has been recently reported that environmental chemical contamination could play a role in the development of this disease. We performed a cross-sectional study in a representative sample from this archipelago to evaluate whether serum levels of selected POPs could be considered as risk factors for diabetes in this population. Serum levels of 30 POPs were determined in 429 adults (9.3% with T2D). We found that serum levels of p,p'-DDE (DDE), PCB-153 and PCB-118 were significantly higher among subjects having diabetes than in non-diabetic subjects (p=0.001, p=0.046, and p<0.0001, respectively). We observed a positive correlation between serum p,p'-DDE and glucose levels. Serum p,p'-DDE was identified as a risk factor for diabetes in univariate analysis in the whole series, and it remained as an independent risk factor for diabetes in subjects with serum glucose <126mg/dL (multivariate analysis, Exp(B)=1.283, CI 95% (1.023-1.611), p=0.031). Those normoglycemic subjects that are most exposed to p,p'-DDE (95th percentile: serum p,p'-DDE>5µg/L) seem to be those people at higher risk. Our results showed that p,p'-DDE levels were significantly higher among subjects having diabetes. These findings should be considered by public health Authorities to implement measures devoted to minimize human exposure to pollutants that could be harmful to the population.

Comparative study of the intake of toxic persistent and semi persistent pollutants through the consumption of fish and seafood from two modes of production (wild-caught and farmed).

Adverse effects of chemical contaminants associated with seafood counteract the undoubted benefits for the health of its valuable nutrients. So much so that many dietary guidelines recommend no more than one serving a week of fish and seafood. Although is estimated that aquaculture provides more than 50% of the fish and seafood consumed globally, few research studies have focused in the assessment of the intake of pollutants through aquaculture products. In this study we determined the levels of organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs), and toxic elements (Pb, Cd, Ni, Al, As, and Hg) in a large sample of farmed and wild-caught seafood, and we estimated the intake of these contaminants in two hypothetical models of consumers: those consuming only farmed fish, and those consuming only wild fish. Measured levels of most organic and many inorganic pollutants were higher in aquaculture products, and consequently intake levels if only such products were consumed would be also significantly higher. Thus, the intake of ∑PAHs in adults consuming aquaculture seafood would be 3.30ng/kg-bw/day, and consuming seafood from extractive fishing 2.41ng/kg-bw/day (p<0.05); ∑OCPs, 3.36 vs. 1.85ng/kg-bw/day, respectively (p<0.05); ∑PCBs, 2.35 vs. 2.11ng/kgbw/day, respectively; and the intake of Pb, Ni, As, and Al would be also significantly higher consuming farmed seafood. For children the estimations were very similar, but the difference of intake of PCBs reached statistical significance. The implementation of several decontamination practices in aquaculture would allow not only match the levels of pollution from wild-caught seafood, but also could provide products with much lower levels of pollutants than those, which in turn would allow to increase consumption over the "one serving per week", and so benefit the consumer of the enormous positive health effects of the valuable nutrients of seafood.

Comparative analysis of selected semi-persistent and emerging pollutants in wild-caught fish and aquaculture associated fish using Bogue (Boops boops) as sentinel species.

The marine environment acts as a sink for diverse anthropogenic pollutants, although the environmental contamination may be non-uniformly distributed. In recent decades, the aquaculture sector has experienced a steady growth postulating as a good alternative for seafood production. However, a social debate exits about the differential level of pollutants in wild and farmed species. This study was designed to evaluate the level of pollutants in a sentinel species: Bogue (Boops boops) associated and non-associated to fish-farm cages. A total of 82 chemical substances were determined by gas chromatography-mass spectrometry, including persistent (polychlorobiphenyls (PCBs) and organochlorine pesticides (OCPs)), semi-persistent (bromodiphenyl ethers (BDEs) and polycyclic aromatic hydrocarbons (PAHs)), and emerging pollutants (such as organophosphate flame retardants (OPFRs) and UV-filters). In general, aquaculture-associated bogues showed lower levels of semi-persistent and emerging pollutants than wild-caught fish, especially when sums were considered. Thus, sum of BDEs was significantly lower in the aquaculture group (p=0.01). A similar trend was also observed for benzo(a)anthracene, the UV-filter 2-ethylhexyl-p-methoxycinnamate and some OPFRs. In the case of persistent pollutants, the sum of dioxin-like PCBs and sum of DDTs were lower in the group of wild-caught bogues (p=0.034 and p=0.003, respectively) than in aquaculture-associated bogues, as previously described for some aquaculture species. Fish feed appear as an important factor in the uptake of such substances suggesting a diet intervention to reduce their levels in the aquaculture products. Another interesting result is that for almost all chemical substances analyzed, bogues captured near sewage outfalls showed the highest levels of pollutants, pointing out the need of stringent measures for wastewater treatment units discharging in coastal areas. On the light of these results, further research in specific farmed and wild fish species in relation to their dietary value and pollutant's levels seems to be mandatory.

The heartworm (Dirofilaria immitis) seems to be able to metabolize organochlorine pesticides and polychlorinated biphenyls: A case-control study in dogs.

It has been described that the co-existence of parasite infection and chemical exposure has various effects on the accumulation of persistent organochlorine pollutants (POPs) in the host. Certain parasites are not only able to accumulate POPs but also seem to have the ability to metabolize certain compounds. We have designed a case-control study aimed to disclose the role of Dirofilaria immitis in the bioavailability of POPs in dogs trying to know whether these parasites store or metabolize the POPs. A total of 40 common POPs (18 polychlorinated biphenyls congeners (PCBs) and 22 organochlorine pesticides were quantified in dog serum. The study included three groups of dogs prospectively recruited in the island of Gran Canaria (Canary Islands, Spain): a) control animals, non-parasitized (serologically tested negative, n=24); b) a group constituted by dogs tested positive for heartworm disease (D. immitis) and negative for other parasites (n=25); and c) the same group of parasitized dogs after the treatment against the parasite (n=25). The presence of D. immitis was strongly associated with lower serum levels of a wide range of pollutant in their hosts (PCB congeners 28, 105, 118, 123, 138, 153, 167 and 180; hexachlorobenzene, lindane, aldrin, dieldrin, and methoxychlor). The serum levels of these substances remained at very low levels after the treatment against the parasite, suggesting that D. immitis do not simply store such compounds, but they probably have some ability to metabolize these pollutants. We encourage the use of the parasite infestation status as a cofactor that needs to be taken into account in studies aimed to evaluate the serum levels of POPs.

Differential gene expression pattern in human mammary epithelial cells induced by realistic organochlorine mixtures described in healthy women and in women diagnosed with breast cancer.

Organochlorine pesticides (OCs) have been associated with breast cancer development and progression, but the mechanisms underlying this phenomenon are not well known. In this work, we evaluated the effects exerted on normal human mammary epithelial cells (HMEC) by the OC mixtures most frequently detected in healthy women (H-mixture) and in women diagnosed with breast cancer (BC-mixture), as identified in a previous case-control study developed in Spain. Cytotoxicity and gene expression profile of human kinases (n=68) and non-kinases (n=26) were tested at concentrations similar to those described in the serum of those cases and controls. Although both mixtures caused a down-regulation of genes involved in the ATP binding process, our results clearly indicate that both mixtures may exert a very different effect on the gene expression profile of HMEC. Thus, while BC-mixture up-regulated the expression of oncogenes associated to breast cancer (GFRA1 and BHLHB8), the H-mixture down-regulated the expression of tumor suppressor genes (EPHA4 and EPHB2). Our results indicate that the composition of the OC mixture could play a role in the initiation processes of breast cancer. In addition, the present results suggest that subtle changes in the composition and levels of pollutants involved in environmentally relevant mixtures might induce very different biological effects, which explain, at least partially, why some mixtures seem to be more carcinogenic than others. Nonetheless, our findings confirm that environmentally relevant pollutants may modulate the expression of genes closely related to carcinogenic processes in the breast, reinforcing the role exerted by environment in the regulation of genes involved in breast carcinogenesis.

Assessment of human health hazards associated with the dietary exposure to organic and inorganic contaminants through the consumption of fishery products in Spain.

In this work we have evaluated the potential carcinogenic and acutely toxic risks associated to the exposure to highly prevalent organic and inorganic contaminants through the consumption of fishery products by the Spanish population. The concentrations of 8 organochlorine pesticides (OCPs), 18 polychlorinated biphenils (PCBs), 7 polycyclic aromatic hydrocarbons (expressed as benzo[a]pyrene toxic equivalents (B[a]Peq)), and three inorganic toxic elements [arsenic (As), cadmium (Cd), and mercury (Hg)] were determined in 93 samples of the most consumed species of white fish, blue fish, cephalopods and seafood species, which were acquired directly in markets and supermarkets in the Canary Islands, Spain. The chemical concentration data were combined with the pattern of consumption of these foodstuffs in order to calculate the daily intake of these contaminants, and on this basis the risk quotients for carcinogenicity and acute toxicity were determined for Spanish adults and children. Our results showed that the daily intake of OCPs, PCBs and B[a]Peq, which is associated to blue fish consumption was the highest within the fish group. The estimated intake of pollutants can be considered low or very low for the individual contaminants, when compared to reference values, except in the case of HCB and As. All the estimated intakes were below the reported Tolerable Daily Intakes. Considering the additive effects of multiple contaminants, the risk of acute toxic effects can also be considered as low or very low. However, our results reflect that the current consumption of white fish in adults and children, and also the blue fish in the case of adults, poses a moderate carcinogenic risk to Spanish consumers, mainly related to their concentrations of As. The conclusions of this research may be useful for the design of appropriate risk communication campaigns.

Different pattern of contamination by legacy POPs in two populations from the same geographical area but with completely different lifestyles: Canary Islands (Spain) vs. Morocco.

The archipelago of the Canary Islands is one of the so-called ultra-peripheral territories of the European Union due to its geographical location away from the continent. Although the level of socioeconomic development and lifestyle of this region is comparable to that of any other of the European Union, it is just 100 km off the coast of Morocco, in the African continent. The population of the Canaries has been extensively studied with respect to their levels of POPs, and it has been described that their levels are relatively high compared to other European regions. It has been speculated with that the proximity to Africa may be associated with this level of contamination, but so far this theory has not been verified. This paper describes for the first time the levels of organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) in a sample of the population of Morocco (n = 131), which were compared with those of a similar sample of the population of permanent residents in the Canary Islands (n = 100) in order to check this hypothesis. Our results showed that Moroccans have higher median values of OCPs than the residents in the Canaries (∑ OCP = 150.2 ng/g lw vs. 83.4 ng/g lw, p = 0.0001). Regarding the PCBs, although recent studies have reported that new environmental sources of PCBs exist in several African countries (including Morocco), the plasma levels of most congeners were significantly higher in Canarians than in Moroccans, especially for the dioxin-like PCBs (median = 7.3 ng/g lw vs. 0.0 ng/g lw, p = 0.0001). The detailed analysis of our results suggests that the levels of these pollutants in the Canarian people are more influenced by their lifestyle and the previous use of these chemicals in the archipelago than by its geographical vicinity with Morocco.

Influence of parasitism in dogs on their serum levels of persistent organochlorine compounds and polycyclic aromatic hydrocarbons.

Persistent organochlorine pollutants (POPs) are toxic chemicals, which accumulate in humans and animals, as only few species have the capability of eliminating them. However, some authors have pointed to the possibility that certain species of invertebrates (i.e. nematodes) could metabolize this type of compounds. As certain species of nematodes act as parasites of vertebrates, this research was designed to explore the influence of some of the most common parasites of the dogs in their serum levels of 56 common POPs. The study included three groups of dogs (n=64), which were prospectively recruited in the island of Gran Canaria (Canary Islands, Spain): a) control animals, non-parasitized (serologically tested negative, n=24); b) dogs tested positive for intestinal parasites and negative for other parasites (n=24); and c) dogs tested positive for heartworm disease (Dirofilaria immitis) and negative for other parasites (n=16). The presence of Dirofilaria immitis was strongly associated with lower serum levels of a wide range of pollutant in their hosts (PCB congeners 28, 52, 118, 138, 153, and 180; hexachlorobenzene, lindane, aldrin, dieldrin, anthracene and pyrene). We also found an inverse association between the hosts' serum levels of PCBs and intestinal parasites. We did not find any association with DDT or its metabolites, but this might be explained by the recently suggested ability of dogs for the efficient metabolization of these compounds. According to the results of this study certain forms of parasitism would reduce the bioavailability of the major classes of POPs in dogs. However, further studies are needed to elucidate whether this phenomenon is due to a competence between parasites and hosts or could respond to a possible capability of parasitic nematodes for the metabolization of these POPs.

Rho GTPases in human cancer: an unresolved link to upstream and downstream transcriptional regulation.

The high incidence of overexpression of some members of the Rho family of GTPases in human tumors suggests that (1) these proteins are involved in cancer onset, and (2) they are potential candidates for a therapeutic intervention. In recent years, the characterization of downstream effectors to Rho GTPases has provided crucial clues on the general cellular effects that permit aberrant proliferation and adhesiveness of tumor cells. The activation of many of these effector proteins in turn results in the modulation of the activity of several transcription factors that play an important role at various levels of Rho signaling. The precise mechanisms by which Rho GTPases participate in carcinogenesis are still not fully understood. However, it is becoming more evident that the specific role of Rho overexpression in tumor initiation, progression and metastasis, as well as the nature and cause of such overexpression in specific human tumors (i.e., transient or stable; tumor environment-regulated; genetic or epigenetic) may be linked to the activation of specific signaling pathways that result in transcriptional regulation. In this review, we summarize the functions of Rho proteins in the regulation of several transcription factors and their relationship to tumor biology.

In vitro evaluation of oestrogenic/androgenic activity of the serum organochlorine pesticide mixtures previously described in a breast cancer case-control study.

Some organochlorine pesticides (OCs) have been individually linked to breast cancer (BC) because they exert oestrogenic effects on mammary cells. However, humans are environmentally exposed to more or less complex mixtures of these organochlorines, and the biological effects of these mixtures must be elucidated. In this work we evaluated the in vitro effects exerted on human BC cells by the OC mixtures that were most frequently detected in two groups of women who participated in a BC case-control study developed in Spain: healthy women and women diagnosed with BC. The cytotoxicity, oestrogenicity, and androgenicity of the most prevalent OC mixtures found in healthy women (H-mixture) and in BC patients (BC-mixture) were tested at concentrations that resembled those found in the serum of the evaluated women. Our results showed that both OC mixtures presented a similar oestrogenic activity and effect on cell viability, but BC-mixture showed an additional anti-androgenic effect. These results indicate that although the proliferative effect exerted by these mixtures on human breast cells seems to depend mainly on their oestrogenic action, the BC-mixture might additionally induce cell proliferation due to its anti-androgenic activity, therefore increasing the carcinogenic potential of this mixture. The findings of this study demonstrate that subtle variations in the composition of a mixture may induce relevant changes in its biological action.

The assessment of daily dietary intake reveals the existence of a different pattern of bioaccumulation of chlorinated pollutants between domestic dogs and cats.

Pet dogs and cats have been proposed as sentinel species to assess environmental contamination and human exposure to a variety of pollutants, including POPs. However, some authors have reported that dogs but not cats exhibit intriguingly low levels of some of the most commonly detected POPs, such as DDT and its metabolites. This research was designed to explore these differences between dogs and cats. Thus, we first determined the concentrations of 53 persistent and semi-persistent pollutants (16 polycyclic aromatic hydrocarbons (PAHs), 18 polychlorinated biphenyls (PCBs) and 19 organochlorine pesticides (OCPs)) in samples of the most consumed brands of commercial feed for dogs and cats, and we calculated the daily dietary intake of these pollutants in both species. Higher levels of pollutants were found in dog food and our results showed that the median values of intake were about twice higher in dogs than in cats for all the three groups of pollutants (ΣPAHs: 274.8 vs. 141.8; ΣOCPs: 233.1 vs. 83; ΣPCBs: 101.8 vs. 43.8 (ng/kg bw/day); respectively). Additionally, we determined the plasma levels of the same pollutants in 42 and 35 pet dogs and cats, respectively. All these animals lived indoors and were fed on the commercial brands of feed analyzed. As expected (considering the intake), the plasma levels of PAHs were higher in dogs than in cats. However, for organochlorines (OCPs and PCBs) the plasma levels were much higher in cats than in dogs (as much as 23 times higher for DDTs), in spite of the higher intake in dogs. This reveals a lower capacity of bioaccumulation of some pollutants in dogs, which is probably related with higher metabolizing capabilities in this species.

An estimation of the carcinogenic risk associated with the intake of multiple relevant carcinogens found in meat and charcuterie products.

Numerous epidemiological studies have demonstrated a link between excessive meat consumption and the incidence of various cancers, especially colorectal cancer, and it has been suggested that environmental carcinogens present in meat might be related to the increased risk of cancer associated with this food. However, there are no studies evaluating the carcinogenic potential of meat in relation to its content of carcinogens. Our purpose was to emphasize the relevance of environmental carcinogens existing in meat as a determinant of the association between cancer and meat consumption. Because within Europe, Spain shows high consumption of meat and charcuterie, we performed this study focusing on Spanish population. Based on the preferences of consumers we acquired 100 samples of meat and charcuterie that reflect the variety available in the European market. We quantified in these samples the concentration of 33 chemicals with calculated carcinogenic potential (PAHs, organochlorine pesticides, and dioxin-like PCBs). The carcinogenic risk of these contaminants was assessed for each food using a risk ratio based on the current consumption of meat and charcuterie and the maximum tolerable intake of these foods depending on the level of contamination by the carcinogens they contain. Our results indicate that the current consumption of beef, pork, lamb, chicken, and "chorizo", represents a relevant carcinogenic risk for consumers (carcinogenic risk quotient between 1.33 and 13.98). In order to reduce carcinogenic risk, the study population should halve the monthly consumption of these foods, and also not to surpass the number of 5 servings of beef/pork/chicken (considered together).

Methodology for the identification of 117 pesticides commonly involved in the poisoning of wildlife using GC-MS-MS and LC-MS-MS.

Poisoning with agricultural chemicals is a major threat for wildlife all over the world. We have developed and validated an analytical scheme aimed to the identification and quantification of a wide range of pesticides in fresh liver from wildlife specimens that are routinely delivered to the forensic laboratories for toxicological investigation. The proposed method is comprised of a general solid-liquid extraction followed by purification steps and three complementary liquid or gas chromatographic analyses with triple quadrupole mass spectrometry detection. The developed methodology allows for the determination of 117 highly toxic pesticides in a variety of samples from wildlife poisoning incidents. The validity of the method has been demonstrated in samples from 98 real cases submitted to our laboratory between 2010 and 2012. This method allowed the identification and quantification of poison in 78 of 94 fresh liver samples from wild animals and was successfully used for the identification of pesticides in 35 of 46 non-liver samples. Therefore, the extraction and cleanup method with minor modifications and the potency of triple quadrupole mass spectrometry allow this method to be used to simultaneously detect and quantify or semi-quantify a majority of the most toxic pesticides in a variety of complex and degraded matrices.