Biomedical subjects

Cinta Porte

Publications and source records attributed to Cinta Porte.

30 records · Page 2Linked to original sources

PubMed2004

Role of B-esterases in assessing toxicity of organophosphorus (chlorpyrifos, malathion) and carbamate (carbofuran) pesticides to Daphnia magna.

In this study, the cladoceran Daphnia magna was exposed to two model organophosphorous and one carbamate pesticides including malathion, chlorpyrifos and carbofuran to assess acetylcholinesterase (AChE) and carboxylesterase (CbE) inhibition and recovery patterns and relate those responses with individual level effects. Our results revealed differences in enzyme inhibition and recovery patterns among the studied esterase enzymes and pesticides. CbE was more sensitive to organophosphorous than AChE, whereas both CbE and AChE showed equivalent sensitivities to the carbamate carbofuran. Recovery patterns of AChE and CbE activities following exposure to the studied pesticides were similar with 80-100% recoveries taking place 12 and 96 h after exposure to organophosphorous and carbamates pesticides, respectively. The physiological role of AChE and CbE inhibition patterns in Daphnia was examined by using organophosphorous and carbamate compounds alone and with specific inhibitors of CbE. Under exposure to organophosphorous pesticides, survival of Daphnia juveniles was impaired at AChE inhibition levels higher than 50% whereas under exposure to the carbamate carbofuran low levels of AChE inhibition affected mortality. Inhibition of CbE by 80-90% increased toxicity to organophosphorous and carbamate pesticides by up to two- and four-fold, respectively. Our results suggest that both AChE and CbE enzymes are involved in determining toxicity of Daphnia to the studied chemicals and that AChE inhibition levels higher than 50% can be considered of environmental concern to Daphnia species.

Animals↗

PubMed2004

Esterification of vertebrate-type steroids in the Eastern oyster (Crassostrea virginica).

Characteristics of acyl-coenzyme A (acyl-CoA):steroid acyltransferase from the digestive gland of the oyster Crassostrea virginica were determined by using estradiol (E2) and dehydroepiandrosterone (DHEA) as substrates. The apparent Km and Vmax values for esterification of E2 with the six fatty acid acyl-CoAs tested (C20:4, C18:2, C18:1, C16:1, C18:0, and C16:0) were in the range of 9-17 microM E2 and 35-74 pmol/min/mg protein, respectively. Kinetic parameters for esterification of DHEA (Km: 45-120 microM; Vmax: 30-182 pmol/min/mg protein) showed a lower affinity of the enzyme for this steroid. Formation of endogenous fatty acid esters of steroids by microsomes of digestive gland and gonads incubated in the presence of ATP and CoA was assessed, and at least seven E2 fatty acid esters and five DHEA fatty acid esters were observed. Some peaks eluted at the same retention times as palmitoleoyl-, linoleoyl-, oleoyl/palmitoyl-, and stearoyl-E2; and palmitoleoyl-, oleoyl/palmitoyl-, and stearoyl-DHEA. The same endogenous esters, although in different proportions, were produced by gonadal microsomes. The kinetic parameters for both E2 (Km: 10 microM; Vmax: 38 pmol/min/mg protein) and DHEA (Km: 61 microM; Vmax: 60 pmol/min/mg protein) were similar to those obtained in the digestive gland. Kinetic parameters obtained are similar to those observed in mammals; thus, fatty acid esterification of sex steroids appears to be a well-conserved conjugation pathway during evolution.

Acyltransferases↗

PubMed2004

Experimental designs to assess endocrine disrupting effects in invertebrates. A review.

In order to gain basic understanding of the ecological effects of vertebrate Endocrine Disrupting Chemicals (EDCs), many research groups are currently testing these chemicals using aquatic invertebrates. Small crustaceans, such as cladocerans and copepods, are of particular interest since they are ecologically important and their short life cycles allow obtaining information on demographic parameters. Despite the existence of diverse literature on the development, growth and reproductive effects of EDCs on these crustaceans, only a few studies have unambiguously assessed a truly endocrine disrupting effect. This review discusses new experimental designs to differentiate between endocrine disruption and other causes of reproductive and developmental impairment. Our findings clearly illustrate that many studies may have falsely concluded that chemicals have endocrine disrupting modes of action when in fact a much simpler explanation was not previously ruled out (e.g., egg mortality, feeding inhibition). This means that there is an urgent need for integration of toxic effects on energy intake to toxicity assessments. Such an approach would permit different ectotoxicological models of action, including endocrine disrupting effects, to be distinguished and their relative roles in the overall toxic response to be clarified.

Animals↗

PubMed2004

Interaction of tributyltin with hepatic cytochrome P450 and uridine diphosphate-glucuronosyl transferase systems of fish: in vitro studies.

Hepatic microsomes of red mullet (Mullus barbatus) and flounder (Platichthys flesus) were preincubated in the presence of a concentration range of the antifouling agent tributyltin (TBT) chloride, and the interactions of TBT with cytochrome P450 and uridine diphosphate-glucuronyl transferase systems were investigated. The enzyme systems were examined in terms of cytochrome P4501A (CYP1A)-catalyzed 7-ethoxyresorufin O-deethylase (EROD) activity and benzo[a]pyrene (BaP) metabolism and in terms of glucuronidation of testosterone and 17beta-estradiol, respectively. Ethoxyresorufin O-deethylase and BaP hydroxylase (BPH) activities of both fish species were progressively inhibited by increasing concentrations of TBT, and the effects were more pronounced for EROD than for BPH (maximal inhibition at 100 microM TBT for EROD and 250-500 microM TBT for BPH). Hydroxylated metabolites of BaP (3-hydroxy-, 7,8-dihydrodiol, and 9,10-dihydrodiol), representing 95% of the total metabolites formed, were reduced up to 75% in the presence of 100 to 500 microM TBT, whereas the formation of other metabolites was less affected. This may alter BaP toxicity and carcinogenicity. Overall, the results were consistent with a specific inhibitory effect of TBT on CYP1A in the two fish species. Additionally, the conjugation of testosterone was significantly inhibited (20%) at low TBT doses (5 microM), with no effect on the glucuronidation of estradiol.

Animals↗

PubMed2003

Long-term exposure effects in vitellogenin, sex hormones, and biotransformation enzymes in female carp in relation to a sewage treatment works.

In the present study, the estrogenic response in the reproductive cycle of female carp, Cyprinus carpio, in relation to a sewage treatment works (STWs) was measured as: alterations in plasmatic and hepatic vitellogenin (VTG) content, plasma levels of testosterone (T) and estradiol (E2), and effects on key enzymes of phases I and II biotransformation, ethoxyresorufin O-deethylase (EROD) and glutathione S-transferase (GST). Low plasma VTG content in females collected 4 km downstream of an effluent discharge suggested possible estrogenic disturbances; however, hepatic VTG content and sex hormone levels did not mirror this. In fact, E2 and T largely fluctuated among fish, with individual variations even greater than among sampling stations or periods. This suggests that at moderately polluted sites, biological variations of female carp and genetic particularities are prevalent. Hepatic biotransformation enzymes, such as EROD, increased with increasing water temperature while, in contrast, GST was maximal at the lowest water temperature.

Animals↗

PubMed2002

Seasonal variation of plasmatic and hepatic vitellogenin and EROD activity in carp, Cyprinus carpio, in relation to sewage treatment plants.

Concern about the health of aquatic fauna living in waters containing biologically active levels of estrogenic compounds is particularly focused on the effects on their reproductive success. To that end, carp, Cyprinus carpio, a feral fish living in warm waters of Southern Europe (NE Spain), were selected for signs of estrogenicity. The study area covered two tributaries (the Anoia and the Cardener) of the Llobregat River both known to be polluted by estrogenic compounds. The estrogenicity in the carp was measured as vitellogenin (VTG) presence in males and alterations in VTG levels in females, over a 6-month period, embracing both the pre- and post-spawning seasons. VTG content was measured in both the plasma and liver, the latter being the organ that synthesizes it. Also, hepatic 7-ethoxyresorufin O-deethylase (EROD) activity was recorded, as interactions of xenoestrogens and oestradiol have been reported to affect this enzymatic activity. The estrogenicity of these rivers was more evident in the Anoia at the location downstream from the sewage treatment plant (STP), by elevated levels of VTG in males and by the presence of some intersex individuals. In the Cardener, no intersex fish were found and male plasmatic VTG was not so highly elevated. However presence of hepatic VTG, in up to 54% of the male fish analyzed, proved exposure to xenoestrogens. In females, VTG fluctuated according to the biological cycle with a plasmatic peak in May and an earlier maximal in the liver. However, this pattern was altered in the locations with higher xenoestrogens presence. EROD activity showed differences between sexes, with higher activity in males than females, as well as site-related differences (up to one order of magnitude) in the same river. These differences were even greater than those detected between rivers. A seasonal trend was also seen in EROD activity with higher induction towards the summer in both males and females.

Animals↗

PubMed2002

Organotin pollution in deep-sea fish from the northwestern Mediterranean.

Aquatic pollution resulting from extensive usage of organotin compounds has been of great concern due to their deleterious effects in nontarget organisms. However, organotin contamination in deep-sea ecosystems has been rarely studied. The presentwork attempted to determine butyltin and phenyltin compounds in deep-sea fish collected between 1000 and 1800 m depth in the NW Mediterranean. The concentration of tributyltin (TBT) and its degradation products, mono- (MBT) and dibutyltin (DBT), aswell astriphenyltin (TPT),and mono- anddiphenyltin (MPT, DPT) was determined in different tissues (liver, gills, digestive tube, and muscle) of several fish species. Total butyltin residues were up to 175 ng/g wet wt, and they were comparable to levels found in coastal fish collected along the Catalan coast. In contrast, deep-sea fish contained much higher levels of phenyltins (up to 1700 ng/g wet wt), and particularly TPT (up to 1430 ng/g wet wt), than previously reported concentrations in shallow-water organisms. The obtained results confirm the long-range transport of organotin compounds to the deep-sea environment, and the subsequent exposure of fish inhabiting nonpoint source areas. The use of TPT in agriculture or as an antifouling agent, its transport to the deep-sea environment associated to particulate matter, and its nonbiodegradable nature in the food chain may account for the high residue levels detected in deep-sea organisms.

Agriculture↗

PubMed2002

The combined use of chemical and biochemical markers to assess water quality in two low-stream rivers (NE Spain).

Carps (Cyprinus carpio) and red swamp crayfish (Procambarus clarkii) were sampled from two lowstream Mediterranean rivers (Anoia and Cardener) receiving extensive urban and industrial waste water discharges. Tissue residues of selected pollutants (organochlorinated compounds, polycyclic aromatic hydrocarbons (PAHs)) were determined in conjunction with different biochemical responses (cytochrome P450, phase II enzymes) with the aim of investigating whether resident organisms were responsive to changes in water quality. Biota inhabiting those rivers were highly exposed to complex mixtures of polychlorobiphenyls and dichlorodiphenyltrichloroethanes (up to 19 ng/g w.w.) and PAHs (up to 6097 ng/g of hydroxylated PAHs in bile), the highest residues being observed in carps from Cardener. This has a reflection on 7-ethoxyresorufin O-deethylase (EROD) activity; that in carps from Cardener ranged between 350 and 550 pmol/min/mg protein, whereas in carps from Anoia ranged between was 90 and 250 pmol/min/mg protein. The highest activity recorded was downstream of the sewage treatment plants in both rivers. Phase II enzymes were less sensitive to polutant exposure than EROD; nonetheless, both glutathione S-transferase and UDP-glucuronyl transferase were higher in carps from Cardener. The results support the usefulness of the combined use of chemical and biochemical responses to assess and diagnose pollution in highly stressed ecosystems.

Animals↗

PubMed2002

Assessment of organotin pollution along the Polish coast (Baltic Sea) by using mussels and fish as sentinel organisms.

Levels of tributyltin (TBT) and its degradation products, mono- (MBT) and dibutyltin (DBT), as well as triphenyltin (TPT), were monitored in 10 stations along the Polish coast (Baltic Sea). Mussel-Mytilus edulis-and fish-Platichthys flesus-were used as sentinel organisms. The bioaccumulation patterns of butyltin and phenyltin compounds varied substantially. Butyltins were detected in mussel tissue from all the sampled stations. Among them, organisms from the Gulf of Gdansk showed the highest residues (68 ng/g w.w. as Sn) in conjunction with elevated TBT/DBT ratios, which suggest recent inputs of TBT in the area. Additionally, flatfish were sampled in the Gulf of Gdansk, and different tissues (liver, digestive tube and gills) were analyzed separately. TPT, although undetected in mussels, was always present in fish. The highest organotin concentration was observed in the liver (369 ng/g w.w. as Sn) of fish caught near Gdansk port. Relatively high concentrations were observed in digestive tube, which points out the ingestion of organotin contaminated food as an important uptake route of those compounds in P. flesus.

Animals↗

PubMed2002

Antioxidant enzymes and peroxisome proliferation in relation to contaminant body burdens of PAHs and PCBs in bivalve molluscs, crabs and fish from the Urdaibai and Plentzia estuaries (Bay of Biscay).

With the aim of studying levels of antioxidant and peroxisomal enzymes and the structure of peroxisomes in relation to body burdens of polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs), mussels Mytilus galloprovincialis, oysters Crassostrea sp., crabs Carcinus maenas and mullets Mugil cephalus were sampled in two Basque estuaries (Bay of Biscay): Urdaibai (Laida, Txatxarramendi, Arteaga, and downstream a sewage treatment plant-STP) and Plentzia. In general, animals showed higher concentrations of contaminants in winter than in summer and no relevant differences were detected among locations. Conversely, antioxidant enzyme activities were higher in summer. Enzyme expression was studied in mullets using immunochemical methods. By immunoblotting season-dependent differences were detected for Mn-superoxide dismutase (Mn-SOD). As for the immunohistochemical staining, mullets sampled in summer in Plentzia showed significantly higher optical densities for acyl-CoA oxidase and lower for both Cu,Zn-SOD and Mn-SOD than those collected downstream a STP as well as higher catalase immunostaining than those collected in winter. Peroxisomal volume density (V(vp)) of mussels sampled in Laida and Txatxarramendi did not show seasonal variations, while for oysters collected in Laida and Arteaga V(vp) was higher in summer. Crab and mullet V(vp) were also higher in summer. In conclusion, the estuaries of Urdaibai and Plentzia can be considered as low to moderately polluted areas and levels of PAHs and PCBs do not show marked variations apart from seasonal variations. Animals can be adapted to low pollution conditions and, under these circumstances, seasonal factors might affect biomarker responses to a greater extent than pollution variations.

Adaptation, Physiological↗

PubMedDate not supplied

Effects of 17beta-estradiol exposure in the mussel Mytilus galloprovincialis.

Mussels Mytilus galloprovincialis were exposed to different concentrations of estradiol (20, 200, and 2000 ng/l) in a semi-static regime (1-day dosing intervals) for up to 7 days in an attempt to see how mussels dealt with exogenous estrogenic compounds. Sex hormone levels were determined in whole tissue. Free-estradiol was only significantly elevated at the highest exposure dose (up to 10-fold). Most of the estradiol was in the tissues as fatty acid esters (> 78%), which sharply increased in a dose-dependent manner (from 4 ng/g in controls to 258 ng/g at the high exposure group). In contrast, neither free nor esterified testosterone levels showed significant differences between control and exposure groups. The results suggest the existence of mechanisms that allow mussels to maintain their hormonal status, and the important role that fatty acid esterification may play within those mechanisms. Synthesis and conjugation rates of estradiol were further investigated by measuring the activity of P450 aromatase, and palmitoyl-CoA:estradiol acyltransferase, in digestive gland microsomal fractions. Overall, the study contributes to the better knowledge of molluscan endocrinology, and defines new mechanisms of regulation of free steroid-levels in mussels.

Acyl Coenzyme A↗

PubMedDate not supplied

Esterification of vertebrate-like steroids in the eastern oyster (Crassostrea virginica).

The esterification of two model vertebrate steroid hormones - estradiol (E2) and dehidroepiandrosterone (DHEA) - was studied in the oyster Crassostrea virginica. The activity of acyl-CoA:steroid acyltransferase was characterized in microsomal fractions isolated from oyster digestive glands. The apparent Km and Vmax values changed with the fatty acid acyl-CoA used (C20:4, C18:2, C18:1, C16:1, C18:0 or C16:0), and were in the range of 9-17 microM, and 35-74 pmol/min/mg protein for E2, and in the range of 45-120 microM, and 30-182 pmol/min/mg protein for DHEA. Kinetic parameters were also assessed in gonadal tissue. The enzyme saturated at similar concentrations, although conjugation rates were lower than in digestive gland. Preliminary data shows that tributyltin (TBT) in the low microM range (1-50) strongly inhibits E2 and DHEA esterification, the esterification of E2 being more sensitive to inhibition than that of DHEA. Overall, results indicate that apolar conjugation occurs in oysters, in both digestive gland and gonads, at a very similar rate to mammals, suggesting that this is a well conserved conjugation pathway during evolution. Esterification, together with other mechanisms, can modulate endogenous steroid levels in C. virginica, and might be a target for endocrine disrupters, such as TBT.

Animals↗