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Biomedical subjects

J M Monserrat

Publications and source records attributed to J M Monserrat.

At least 19 recordsLinked to original sources

Estradiol valerate and tibolone: effects on memory.

This study investigated the effects of estradiol valerate (EV) and tibolone (TB) treatments on some memory parameters of ovariectomized young (2 months), adult (8 months) and old (20 months) female rats. A Sham-operated group was used as control and the animals were treated daily, by oral gavage, with saline (Sham and placebo NR group), EV (0.3 mg/kg) or TB (0.5 or 1 mg/kg, TB1 and TB2, respectively). In step-down inhibitory avoidance task, the latency of old TB2-treated females in the short-term test was significantly inferior (p<0.05), compared to TB2 adults. In the elevated plus maze, adult NR females spent significantly less time (p<0.05) in the open arms as compared with EV and TB2-treated animals. Additionally, adult TB2-treated females spent significantly less time in the closed arms compared to Sham, NR and TB1 groups. Finally, in the water maze retention test, young TB1-treated animals performed worse when compared to Sham, EV and TB2 females. In the old animals, EV treatment hampered subject performance as compared to all other treatments. Taken together, these results indicate that ovarian hormones differently affect female memory in an age-dependent manner.

Aging↗

Importance of cholinesterase kinetic parameters in environmental monitoring using estuarine fish.

The aim of the present study was to determine the kinetic parameters and cholinesterase (ChE) activity in two teleost fish, the white mouth croaker Micropogonias furnieri (Scianidae) and the Madamango sea catfish Cathorops spixii (Ariidae), to verify their suitability as sentinel species of aquatic pollution by anticholinergic compounds. Individuals of each species were captured in one reference and one polluted site in the Southern Brazilian coast. Brain tissue was used as enzyme source. Inhibition kinetic parameters indicated that ChE from C. spixii collected at the reference site showed more affinity (Ka) for eserine that from those collected at the polluted site. The opposite was observed for the carbamylation constants (Kc). Thus, similar inhibition constants (Ki) were observed. M. furnieri brain showed an extremely low sensitivity to in vitro inhibition by eserine, indicating that it is not a suitable biomarker to be employed in environmental monitoring of anticholinergic compounds. Results from the present study also point to the need for considering kinetic analysis when cholinesterase activity is employed as a biomarker.

Animals↗

Effects of microcystins over short- and long-term memory and oxidative stress generation in hippocampus of rats.

Microcystins produced by cyanobacteria are potent inhibitors of some protein phosphatases, but recent evidence also indicates its potential to generate oxidative stress. In the present study, the effects of microcystin raw extracts (Mic; 0.01 and 20microg/L) and purified okadaic acid (OA; 0.01 and 10microg/L) on short- and long-term memory alteration and antioxidant and oxidative damage were investigated in hippocampus of rats. The results showed an amnesic effect with 0.01 and 20microg/L Mic on retrieval and only with 0.01microg/L Mic on spatial learning. Parallel to these effects oxidative damage was observed as evidenced by augmented levels of lipid peroxides and DNA damage and the absence of antioxidant responses in terms of total oxyradical scavenging capacity. Phase II reactions catalyzed by glutathione-S-transferase were not modified after microcystins exposure. Overall this study showed physiological events (retrieval and spatial learning) that can be related to the classical toxic effects of microcystins (i.e., phosphatase inhibition). In addition, evidence of alternative toxicity mechanisms via oxidative stress generation was also obtained. The fact that organic anion transporter polypeptides (OATP) involved in microcystins uptake are expressed not only in liver but also in brain points to the environmental relevance of the observed effects.

Animals↗

Biochemical and physiological responses after exposure to microcystins in the crab Chasmagnathus granulatus (Decapoda, Brachyura).

Microcystins are usually the predominant cyanotoxins present in both drinking and recreational waters after cyanobacterial blooms. Their classic toxic effect is hepatotoxicity through inhibition of serine/threonine phosphatases. However, recent studies also reported oxidative stress generation and disruption of ion regulation in aquatic organisms after microcystins exposure. In the present study, aqueous extracts of Microcystis aeruginosa were administered to the estuarine crab Chasmagnathus granulatus (Decapoda, Brachyura) by gavage in variable doses (from 34 to 860 microg kg(-1)) and exposure times (6, 12, and 72 h). A control group was exposed to saline solution. Analyzed variables included oxygen consumption, lipid peroxidation (LPO), enzyme activities (glutathione S-transferases or GST; alanine aminotransferase or ALT; aspartate aminotransferase or AST; and lactate dehydrogenase or LDH), glycogen, and microcystins content. Oxygen consumption increased in organisms exposed for 12h to 860 microg kg(-1) of microcystins and a similar result was observed after 72 h at doses equal to or higher than 34 microg kg(-1). LPO levels increased in doses equal to or higher than 34 microg kg(-1) after 72 h. GST and LDH activities increased after 12 h (at a dose of 860 microg kg(-1)), but ALT and AST activities remained unaltered in all experimental conditions. Glycogen content decreased after 72 h exposure at doses equal to or higher than 172 microg kg(-1). After 12h of exposure to 860 microg kg(-1) of microcystins, the concentration found in the hepatopancreas of C. granulatus was 13.17+/-0.56 microg kg(-1). In crabs exposed to doses higher than 172 microg kg(-1) during 72 h this value raised to 32.14+/-4.12 microg kg(-1). The obtained results indicated that microcystins exposure led the tissue to an oxidative stress condition (high LPO levels), at least in part favored by the augment of oxygen consumption, altering the glycogen metabolism. GST responses were only observed in the short-term experiment (12 h) and no effect on classical markers of vertebrate liver damage (ALT and AST) was observed. Although the hepatopancreas from C. granulatus accumulated a relatively low concentration of toxins, it was enough to induce physiological and biochemical disturbances.

Animals↗

Antioxidant responses after microcystin exposure in gills of an estuarine crab species pre-treated with vitamin E.

Microcystins are hepatotoxins suspected to generate oxidative stress. This mechanism was evaluated in gills of the estuarine crab Chasmagnathus granulatus (Decapoda, Brachyura). Adult male crabs were fed ground beef with or without vitamin E (600 mg/kg). Microcystin (1.21 microg/kg) was daily administered through forced ingestion, for 7 days. After exposure, catalase activity was reduced in posterior gills of crabs supplemented with vitamin E. A lower increment in glutathione S-transferase activity (GST) was observed in organisms pretreated with vitamin E and then exposed to microcystin with respect to those exposed to the toxin but not pretreated with the vitamin. Pretreatment with vitamin E also increased nonproteic sulfhyrdil groups and this effect was not observed after microcystin exposure. The fact that supplementation with antioxidants such as vitamin E modulates GST activity indicates the direct or indirect involvement of microcystin in oxidative stress generation.

Animals↗

Antioxidant responses and oxidative stress after microcystin exposure in the hepatopancreas of an estuarine crab species.

Antioxidant responses and oxidative stress were evaluated in the hepatopancreas of the estuarine crab Chasmagnathus granulatus (Decapoda, Brachyura) after oral microcystin administration. Responses were evaluated through antioxidant enzyme activities (catalase-(CAT), superoxide dismutase, glutathione-S-transferase- (GST)). Nonproteic sulfhydril (NP-SH) groups, oxygen consumption, lipid peroxides (LPO), and oxidized proteins were also measured. Microcystin administration increased the oxygen consumption. GST activity and NP-SH concentration showed transient increases and CAT activity showed a peak and then a reduction. Oxidative damage was evidenced with regard to LPO content and suggested by the inhibition of CAT activity at the end of the experiment, indicating that the antioxidant response induced by the toxin was insufficient. A lowering in the number of hepatopancreatic B cells should be related to microcystin elimination.

Animals↗

Oxidative stress responses in two populations of Laeonereis acuta (Polychaeta, Nereididae) after acute and chronic exposure to copper.

Acute (4 days) and chronic (14 days) effects of copper were evaluated on the antioxidant defenses of Laeonereis acuta (Polychaeta) collected in unpolluted (UP) and polluted (P) sites. In the acute assay (125 and 250 micro g Cu/l) superoxide dismutase (SOD) and glutathione S-transferase (GST) activities did not change, whereas catalase (CAT) increased in worms from both the sites. Lipid peroxidation was higher in copper exposed worms from the P site. In the chronic assay (62.5 micro g Cu/l) polychaetes from the P site showed enhanced activities of SOD, GST and CAT and higher contents of metallothionein-like proteins and sulfhydrils compared to worms from UP. Differences in responses between polychaetes from UP and P sites suggest that organisms from the polluted site, P, are more susceptible to oxidative stress conditions.

Analysis of Variance↗

Determination of lipid peroxides in invertebrates tissues using the Fe(III) xylenol orange complex formation.

Reactive oxygen species (ROS) are subproducts of the oxidative metabolism known to initiate chain reactions with polyunsaturated fatty acids that generate lipid peroxides (LPO). The objective of this work was to adapt the ferrous oxidation/xylenol orange (FOX) assay to measure LPO in invertebrate tissues i.e.: from polychaeta (Laeonereis acuta) and crab (Chasmagnathus granulata) species. Whole polychaetes were homogenized in methanol 100%, being determined the optimal sample volume and the time required for color development. It was tested five sample volumes (8-30 microl), following color development up to 215 min. Absorbance stabilization was observed after 90 min, being linearly related with sample volume. A similar procedure was adopted for crab tissues (anterior gills, posterior gills, and hepatopancreas). Differences between species and between organs of the same species were observed when analyzed nonspecific absorbance increments after adding the standard cumene hydroperoxide (CHP). In polychaeta and crab anterior gills tissue, absorbance increments were lower (21-25%) than samples without tissue extracts (blanks) that received CHP. In crab posterior gills and hepatopancreas, the nonspecific increment was almost negligible. Correction formulae are given to account for these differences and simplified protocols for each tissue and species are also included. Great differences in the lipid peroxides content was detected between worms (127.05 +/- 19.32 nmoles CHP/g of wet tissue) respect to anterior gills, posterior gills, and hepatopancreas from the crab species (52.65 +/- 3.59, 30.54 +/- 4.73, and 48.51 +/- 8.78 nmoles CHP/g of wet tissue, respectively).

Animals↗

Effect of microcystin on ion regulation and antioxidant system in gills of the estuarine crab Chasmagnathus granulatus (Decapoda, Grapsidae).

The objective of this work was to evaluate mechanisms of microcystin toxicity on crustacean species. Adult male crabs of Chasmagnathus granulatus (13.97+/-0.35 g) acclimated to low salinity (2 per thousand ) were injected with saline (control) or Microcystis aeruginosa aqueous extract (39.2 microg/l) at 24 h intervals for 48 h. After the exposure period, the anterior and posterior gills were dissected, measuring Na(+),K(+)-ATPase and glutathione-S-transferase (GST) activity. Total oxyradical scavenging capacity (TOSC) and lipid peroxides (LPO) content were also determined. Na(+),K(+)-ATPase activity in anterior gills was significantly lower in crabs injected with toxin than in control crabs, while no significant difference in the enzyme activity was detected in posterior gills. Both sodium and chloride concentration in the hemolymph were not affected by toxin exposure. Significant changes in GST activity were detected in posterior gills, with higher values being observed in the toxin-injected crabs. Crabs exposed to microcystin also showed a significant increase in the TOSC value against peroxyl radicals, for both anterior and posterior gills. Lipid peroxides level did not change in both gill types after exposure to the toxin. The increased levels of TOSC suggest the occurrence of a crab response against oxidative stress induced by toxin injection, which prevents lipid peroxidation.

Animals↗

Toxic effects of microcystins in the hepatopancreas of the estuarine crab Chasmagnathus granulatus (Decapoda, Grapsidae).

Microcystins are toxins produced by cyanobacteria, being toxic to aquatic fauna. It was evaluated alternative mechanisms of microcystins toxicity, including oxidative stress and histopathology in the hepatopancreas of the estuarine crab Chasmagnathus granulatus (Decapoda, Grapsidae). Microcystins was administered to crabs (MIC group) over 1 week, whereas the control (CTR group) received the saline from cyanobacteria culture medium. At day 7, catalase activity was higher in the MIC than in the CTR group, although a decrease of activity was verified in both groups with respect to time 0. Glutathione-S-transferase activity augmented in MIC with respect to CTR, suggesting a higher conjugation rate of the toxins with glutathione. No differences were detected in the superoxide dismutase activity. Lipid peroxidation remained stable in both groups. Histopathological analyses showed that the number of B cells decreased significantly in the CTR as a possible effect of starvation, while no significant change was observed in the MIC group. The hepatopancreas from the MIC group exhibited some necrotic tubules and melanin-like deposits. Overall, results showed that some enzymes of the antioxidant defense system were activated after microcystins exposure, this response being able to maintain lipid peroxidation levels, but insufficient to completely prevent histological damage.

Animals↗

Acute and chronic effects of cadmium on blood homeostasis of an estuarine crab, Chasmagnathus granulata, and the modifying effect of salinity.

Whole body oxygen consumption and some hemolymph parameters such as pH, partial pressure of gases, level of ions and lactate were measured in the estuarine crab Chasmagnathus granulata after both acute (96 h) and chronic (2 weeks) exposure to cadmium at concentrations ranging from 0.4 to 6.3 mg/l. In all instances, the crabs developed hemolymph acidosis, but no respiratory (increased PCO2) or lactate increases were evident. Hemolymph levels of sodium and calcium were always increased by cadmium exposure. The chronic toxicity of cadmium was enhanced at 12 per mil salinity, even causing a significantly higher mortality in comparison with the higher salinity (30 per mil ) used. A general metabolic arrest took place at 12 per mil salinity in the crabs chronically exposed to cadmium, as indicated by decreases of oxygen consumption and PCO2, an increase of PO2, along with no changes in lactate levels. These imbalances were associated with severe necrosis and telangiectasia in the respiratory gills, probably leading to respiratory impairment and finally histotoxic hypoxia and death of the animals.

Animals↗

Effects of Anabaena spiroides (Cyanobacteria) aqueous extracts on the acetylcholinesterase activity of aquatic species.

The effects of aqueous extracts from a cyanobacteria species, Anabaena spiroides, on fish (Odontesthes argentinensis), crab (Callinectes sapidus), and purified eel acetylcholinesterase (AChE) activity were studied. In vitro concentrations of A. spiroides aqueous extract that inhibited 50% of enzyme activity (IC50) were 23.0, 17.2, and 45.0 mg/L of lyophilized cyanobacteria for eel, fish, and crab AChE, respectively. Eel AChE inhibition follows pseudo-first-order kinetics, the same expected for organophosphorus pesticides. Inhibition of purified eel AChE using mixtures of bioxidized malathion and aqueous extract of A. spiroides showed a competitive feature (p < 0.05), suggesting that the toxin(s) could be structurally similar to an organophosphorus pesticide and that toxins present in the aqueous extract inhibit the active site of the enzyme. The inhibition recovery assays using 2-PAM (0.3 mM) showed that (1) bioxidized malathion inhibited 27.0 +/- 1.1% of crab and 36.5 +/- 0.1% of eel AChE activities; (2) with bioxidized malathion + 2-PAM the registered inhibition was 13.2 +/- 2.1% and 3.7 +/- 0.5% in crab and eel AChE, respectively; (3) the aqueous extract from A. spiroides inhibited 17.4 +/- 2.2% and 59.9 +/- 0.5% of crab and eel AChE activity, respectively; and (4) aqueous extract + 2-PAM inhibited 22.3 +/- 2.6 and 61.5 +/- 0.2% of crab and eel AChEs. The absence of enzyme activity recovery after 2-PAM exposure could imply that the enzyme aging process was extremely quick.

Acetylcholinesterase↗

Inhibitory effects of cadmium on carbonic anhydrase activity and ionic regulation of the estuarine crab Chasmagnathus granulata (Decapoda, Grapsidae).

This work was aimed at evaluating the gill carbonic anhydrase (CA) activity of the estuarine crab Chasmagnathus granulata exposed in vivo to cadmium, at different salinities. The in vivo effect of the specific inhibitor acetazolamide (AZ) was also assayed. Besides, the inhibition of CA activity by different heavy metals (cadmium, copper, zinc) and AZ were evaluated under in vitro conditions. For the in vivo assays, adult males were acclimated to salinities of 2.5 or 30/1000. The corresponding 96-h LC50 of cadmium was 2.69 mg l-1 at 2.5/1000, and > 50 mg l-1 at 30/1000. Cadmium only caused a significant lower CA activity than control at 2.5/1000. EC50 for CA inhibition was estimated to be 1.59 mg l-1 at 2.5/1000. Statistical differences in Na+ hemolymphatic levels (P < 0.05) were only detected at 2.5/1000, between 0 and 1.25 mg l-1 of cadmium, but no statistical differences were observed for Cl- levels at any assayed salinity. As CA inhibition registered at 2.5/1000 was followed by only changes in Na+ concentration, it is likely that cadmium exposure could differentially affect ions permeability, among others factors. The concentrations that inhibited in vitro 50% of enzymatic activity (IC50) were 2.15 x 10(-5), 1.62 x 10(-5), 3.75 x 10(-6) and 4.4 x 10(-10) M for cadmium, copper, zinc and AZ, respectively. The comparison with IC50 values of other aquatic species, indicates a higher CA sensitivity for C. granulata to pollutants.

Acclimatization↗

Some kinetic and toxicological characteristics of thoracic ganglia cholinesterase of Chasmagnathus granulata (Decapoda, Grapsidae).

In this study, some kinetic and toxicological parameters of the thoracic ganglia cholinesterase from the estuarine crab Chasmagnathus granulata were determined. Effects of the type and concentration of substrate, pH (6.80-8.50), incubation temperature (5-35 degrees C) and eserine on the enzyme activity were studied. Enzymatic activity was higher at pH 7.40 and 8.00 and significantly reduced at lower temperatures (5-10 degrees C). Employing acetylthiocholine iodide (ATCh) as substrate, the K(m) and Vmax were estimated as 0.28 mM and 1.75 mumol.mg protein-1.min-1, respectively. A 14.11 and 24.51% decrease in enzyme activity were registered at 4.62 and 9.24 mM of ATCh, respectively. Using propionylthiocholine iodide as substrate, the K(m) and Vmax were estimated as 0.16 mM and 0.91 mumol.mg protein-1.min-1, respectively. The IC50 for eserine was estimated as 5.3 x 10(-4) mM. The Ki estimated for eserine (8.10 mM-1.min-1) indicates that the thoracic ganglia cholinesterase from C. granulata showed a low ability to generate an irreversible enzyme-inhibitor complex. The higher enzymatic activity registered with ATCh and the enzyme inhibition observed at high concentration of this substrate, suggest that thoracic ganglia cholinesterase from C. granulata is an acetylcholinesterase.

Acetylthiocholine↗

Chronic toxicity of ethyl parathion and isobutoxyethanol ester of 2,4-dichlorophenoxyacetic acid to estuarine juvenile and adult crabs.

The chronic toxicity of ethyl parathion and isobutoxyethanol ester of 2,4-dichlorophenoxyacetic acid was determined for the crabs Uca uruguayensis and Chasmagnathus granulata collected in Argentina in 1989. Parathion toxicity was more time-dependent than 2,4-D. Four-week LC50 values for parathion were size and species independent, as expected under chronic exposure conditions. As shown in previous studies, the opposite was noted when crabs were exposed to acute exposure. Parathion was much more toxic than 2,4-D, in all cases. Also, parathion exerted a very much lower chronic lethal effect than other pesticides at a fraction of the respective acute LC50. A different susceptibility between sexes was found in C. granulata exposed to 2,4-D.

2,4-Dichlorophenoxyacetic Acid↗

Acute and chronic effects of parathion and 2,4 D on the oxygen consumption of Chasmagnathus granulata (Decapoda, Brachyura).

The effect of two pesticides widely used in Argentina on the oxygen consumption of the estuarine crab Chasmagnathus granulata was studied. Constant pressure respirometers were employed to estimate the rate of oxygen consumption per weight unit of animals treated previously with each pesticide, both acute (96 h) and chronically (15 and 30 days). Crabs exposed to parathion -an organophosphorate insecticide that causes the inhibition of acetylcholinesterase- show an increase of oxygen consumption at 0.5 ppm under acute exposure, and at 10 ppb under a chronic one. On the other hand, crabs exposed to 2,4 D (an herbicide) did not show changes in their consumption after an acute exposure, but those exposed chronically did show an increase at low concentration (5 ppm) followed by a relative decrease at the highest concentration (50 ppm). The results obtained for parathion are in accordance with the abnormal cholinergic excitation that it may exert on crustacean nervous system. The effect of 2,4 D was consistent with its uncoupler action at respiratory chain level, at low concentrations, while a possible Krebs cycle enzymes inhibition might be occurring at higher concentrations of that pesticide, as in other crustacean species. The faster action of parathion, respect to 2,4 D, is explained by its neurotoxic nature.

2,4-Dichlorophenoxyacetic Acid↗

[Contribution to the diagnosis of Dubin-Johnson syndrome].

Eight patients with Dubin-Johnson's syndrome have been reported. It appoints the greatest casuistry about the theme in our country. The coincidence covers all the aspects of the disease except by the fact that we emphasize, as a matter of interest, that we were able to visualize the gallblader and the biliary tract through oral cholecystography visualize the gallblader and the biliary tract through oral cholecystography or through intravenous cholangiography, in all the patients who have been studied. It doesn't agree with the class descriptions in which these exams have negative results. So we point out the possibility of this disease's existence in spite of the radiographic visulaization of biliary tract, particularly in those patients with moderated hepatic pigment-deposit. Refering to pathologic anathomy we remark the importance in the search of the pigment charactiristic of the Dublin-Johnson's syndrome in all the liver histopathologic exams. Besides, we consider it valuable to make the Fontana-Masson coloration in those patients who present negative pigment reaction to iron. The systematic examinations of the cytoplasmatic pigments will allow us to diagnose the disease even in the cases with minimum and latent characteristics.

Adolescent↗