Assessment of polycyclic aromatic hydrocarbons (PAHs) in mediterranean top marine predators stranded in SE Spain.

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental pollutants. Although they are not bioaccumulated in vertebrates, chronic exposures might still derive on serious toxic effects. We studied concentrations of 16 reference PAHs on blubber of two dolphin species (striped dolphin, n = 34; and bottlenose dolphin, n = 8) and one marine turtle (loggerhead turtle, n = 23) from the Mediterranean waters of SE Spain, an important or potential breeding area for these and other related species. Σ16 PAHs concentrations were relatively similar between the three species, but they were in the lower range in comparison to worldwide data. Of the six PAHs detected, fluoranthene was the only high molecular weight (HMW) PAH, so low molecular weight (LMW) PAHs predominated. Naphthalene and phenanthrene were invariably those PAHs with higher detection rates as well as those with higher concentrations. In accordance with the literature, sex and length did not have significant influence on PAHs concentrations, probably due to high metabolization rates which prevent for observation of such patterns. Despite LMW PAHs are considered less toxic, we cannot dismiss toxic effects. This is the first work assessing PAHs concentrations in cetaceans and sea turtles from the SE Spain, which could serve as the baseline for future research.

POPs concentrations in cetaceans stranded along the agricultural coastline of SE Spain show lower burdens of industrial pollutants in comparison to other Mediterranean cetaceans.

Despite the Mediterranean Sea being one of the world's marine biodiversity hotspots, it is a hotspot of various environmental pollutants. This sea holds eight cetacean with resident populations whose numbers are considered to decline in the last decades and which are particularly susceptible to POPs bioaccumulation due to their peculiar characteristics. In this work, we studied blubber concentration of various OCPs and several PCBs and PBDEs congeners in cetaceans stranded in the northern coast of the Gulf of Vera (Region of Murcia, SE Spain) between 2011 and 2018. Most compounds and congeners were above the limit of detection in most samples, although some pesticides like endosulfan stereoisomers or endrin were never detected. DDT and its metabolites, PCBs and metoxychlor appear as the dominant compounds while PBDEs shows concentrations of lower magnitude. Striped dolphin was the species accounting for higher concentrations of most pollutants. There were differences in concentrations and profiles between species which could be partially explained by differences on diet and feeding behavior. We also observed differences based on life history parameters suggesting maternal transfer for most POPs, in accordance with other works. DDE/ ΣDDT ratio suggest no recent exposure to these pesticides. Despite showing lower concentrations than some previous works, PCB concentrations accounted for higher total TEQ than many studies. According to toxicity thresholds in the literature, we cannot guarantee the absence of health consequences on populations studied, especially for those caused by PCBs. These findings are of major importance considering the relevance of the study area in the conservation of Mediterranean cetaceans.

Human biomonitoring of persistent organic pollutants in elderly people from the Canary Islands (Spain): A temporal trend analysis from the PREDIMED and PREDIMED-Plus cohorts.

The population of the Canary Islands has been exposed to high levels of persistent organic pollutants (POPs). Biomonitoring studies are essential to know the temporal trend in residue levels, particularly of substances banned decades ago. The purpose of this study was to analyze the distribution of plasma concentrations of 59 POPs in 175 participants from the PREDIMED-Plus trial (2014-2016), and to compare them with the distribution of these POPs in 343 participants in the PREDIMED trial (2006-2009). All participants had metabolic syndrome. No difference in the distribution of age, gender or BMI was observed between trials. POPs were determined by gas chromatography-mass spectrometry. Density plots -POP Geoffrey Rose curves- were used to represent the full population distribution of each compound. Three out of 59 POPs were detected and quantified in ≥95% of the samples (p,p'-DDE, median = 694.7 ng/g lipid; HCB, median = 57.0 ng/g lipid; and ß-HCH, median = 75.7 ng/g lipid). PCB congeners 138, 153 and 180 were detected in 64.6, 40.0 and 88.0% of the samples. Females showed highest concentrations of organochlorine pesticides and those subjects who lost ˃ 5 kg showed significant higher plasma concentrations of POPs. In a range of 6-14 years, plasma concentrations of POPs decreased 3.3-21.6 fold, being notable the decrease of 28.7-fold observed for HCB among women. Despite this sharp decline, levels of POPs are still higher than those reported in other regions, since one third of the subjects included in the present report had high concentration of more than three pollutants. Continuous biomonitoring studies are required to know the evolution of the levels of residues and to evaluate the effectiveness of environmental policies.

Biomonitoring of 45 inorganic elements measured in plasma from Spanish subjects: A cross-sectional study in Andalusian population.

Heavy metals and other toxic elements are frequently detected in humans. Rare earth elements (REE) have arisen as a novel group of substances considered as emerging pollutants due to its dependence for high tech industry. We designed a study aimed to conduct the biomonitoring a total of 45 inorganic elements in the population of Andalusia (Spain). A total of 419 participants were recruited and their plasma samples analyzed. Concentration of elements, including elements in the ATSDR's priority pollutant list and REE were measured by ICP-MS in the blood plasma of participants. Arsenic, copper, lead, selenium, antimony, strontium, and bismuth were detected in ˃98% of subjects. Median values of arsenic, mercury and lead were 1.49, 1.46, and 5.86 ng/mL, respectively. These concentrations did not exceed reference values published by international agencies. We observed a positive correlation between age and plasma concentrations of arsenic, mercury, antimony and strontium. Sum of elements was lower in the group of subjects younger than 45 years old (P = 0.002). Positive correlations were observed between body mass index (BMI) and plasma concentrations of barium, cerium, osmium, tin, and ytterbium. 7 out of 26 REEs showed a percentage of detection ≥ 90%. Bismuth, yttrium, and cerium were quantified at the highest concentrations (median value = 7.7, 0.19, and 0.16 ng/mL, respectively). We found that plasma levels of 6 REEs were higher among males, and a positive correlation between REEs and age was detected. The present results suggest a potential interaction with the human physiology that deserves additional research. Given the high persistence of these elements in the environment, and the significant technological dependence on them, future studies are needed to elucidate the potential sources of exposure and possible adverse effects on health, especially in the most vulnerable populations.

Ethanol levels in legally autopsied subjects (2016-2017): Update of data and epidemiological implications in relation to violent deaths in Canary Islands (Spain).

The aim of this study was to analyze the prevalence of ethanol in individuals (277) subjected to a mandatory medico-legal autopsy in the Institute of Legal Medicine of Las Palmas (Canary Islands, Spain) during 2016-2017, comparing the results with data published in 2015. Blood and/or vitreous humor samples were analyzed by gas chromatography. 31.8% of the individuals were positive to ethanol. We observed a decrease in the prevalence of ethanol among males (p = 0.002). While the prevalence of ethanol was reduced, the concentration among those positive subjects has increased. 11.9% of the series died in a traffic accident and the percentage of positive to ethanol decreased in relation to 2015 (64.3, 25.0, and 35.3%). The number of suicides remains constant, although the age of the suiciders has been significantly reduced (p = 0.022). The results of this study indicate that ethanol is still heavily involved in non-natural deaths.

Evaluation of the applicability of BI-RADS® MRI for the interpretation of contrast-enhanced digital mammography. / Evaluación de la aplicabilidad del léxico BI-RADS® de la resonancia magnética para la interpretación de la mamografía digital con contraste.

OBJECTIVE: To determine the usefulness of BI-RADS® MRI for the morphological description and categorization of images obtained with contrast-enhanced digital mammography in comparison with the final diagnosis. MATERIAL AND METHODS: We included patients who had palpable breast lesions, those who needed clarification regarding abnormalities identified with another imaging technique, and those which needed a complementary evaluation of heterogeneously dense breast parenchyma. Three radiologists working independently used a template with the BI-RADS® MRI qualitative descriptors to evaluate the breast lesions studied with contrast-enhanced digital mammography. In a second phase, two other experienced radiologists reached a consensus about discrepant interpretations. Readers also classified each lesion (both benign and malignant lesions) on the BI-RADS® scale (1 - 5). All the results were compared with the real state of disease (determined by the appropriate gold standard for each type of lesion), and the statistical significance was assessed with the chi-square test. RESULTS: A total of 218 benign lesions and 426 malignant lesions were included. The interobserver agreement among the three radiologists was high (Fleiss-Cohen kappa=0.805; 95% CI: 0.728-0.837). Similarly to what has been reported about breast MRI, on contrast-enhanced digital mammography, malignant lesions with mass effect tended to have an irregular shape with spiculated or ill-defined margins and a pattern of intense and heterogeneous enhancement (p <0.001). Nevertheless, unlike on breast MRI, ring enhancement was not an independent criterion of malignancy on contrast-enhanced digital mammography. For lesions without a mass effect, the only significant descriptor was the intensity of contrast material uptake (p <0.05). Applying the BI-RADS® MRI material to contrast-enhanced digital mammography images enabled the correct classification of 85% of lesions in the benign categories (BI-RADS 1 and BI-RADS 2 and 3 lesions) and of 93% of the lesions in the malignant categories (BI-RADS 4-5); these values are similar to those reported for breast MRI. CONCLUSIONS: The morphological descriptors used in BI-RADS® MRI can be applied to the morphological analysis of breast lesions studied with contrast-enhanced digital mammography. The partial discrepancies in the interpretation did not influence the final BI-RADS® score, and the score enabled good differentiation between benign and malignant lesions.

Factors affecting the precision of lesion sizing with contrast-enhanced spectral mammography.

AIM: To evaluate the precision of the pre-surgical measurement of the size of breast cancer by contrast-enhanced spectral mammography (CESM). MATERIAL AND METHODS: This was a retrospective study of 204 breast cancers. Variables related to tumour biology and anthropometric variables were recorded and considered when evaluating the efficacy of CESM in predicting tumour size. Microscopic measurement of the largest diameter of the tumour at pathology was chosen as the reference standard. RESULTS: The mean size of tumours at pathology was 20.7±15.8 mm, while at CESM it was 23.6±16.7 mm (Bland-Altman 2.9 mm overestimation, 2.9 mm; 95% confidence interval [CI]: -10.3-16.2 mm). Spearman's correlation coefficient was 0.83 (p<0.0001). The concordance analysis indicated that 37.8% of the measurements were concordant, 47% were overestimated, and 15.2% were underestimated. Tumour size, nodal involvement, breast density, and breast size significantly modified the sizing accuracy. CONCLUSION: Quality of tumour size prediction with CESM is good, and this appears to be a promising imaging technique in the surgical planning of breast cancer. Biological tumour features, and anthropological characteristics of the patients do, however, affect the diagnostic precision and should be taken into account.

The impact of red and processed meat consumption on cancer and other health outcomes: Epidemiological evidences.

Meat is one of the staples of the human diet, which provides high-quality nutrients, but that also constitutes a relevant source of cholesterol and saturated fatty acids. Epidemiologic studies have linked consumption of red or processed meat with obesity, type 2 diabetes, cardiovascular diseases, and cancers. Most epidemiological studies suggest that a high intake of meat, especially processed meat, is associated with increased colorectal cancer risk. Potential reasons for the association between high meat intake and colorectal cancer risk include some chemicals naturally contained in meat, or generated by the processing and cooking. From the literature it can be concluded that there is enough epidemiological evidence linking processed meat intake and colorectal cancer risk, but there is limited evidence regarding unprocessed red meat intake and the disease. On the contrary, there is only limited evidence linking meat intake with other diseases such as cardiovascular disease, type 2 diabetes or other cancers. Nevertheless, the literature suggest that dietary intervention may be a promising approach for prevention of cancers of the colon, esophagus, liver, stomach and bladder, and type 2 diabetes and cardiovascular disease which still need to be confirmed by further well designed prospective studies and experimental research.

Organochlorine Pesticides Exposure and Bladder Cancer: Evaluation from a Gene-Environment Perspective in a Hospital-Based Case-Control Study in the Canary Islands (Spain).

The incidence of bladder cancer has increased significantly since the 1950s. Pesticide exposure has been linked with increasing bladder cancer incidence, although the evidence is inconclusive. However, most epidemiological studies did not evaluate the potential role played by the organochlorine pesticides, the most widely used pesticides in Western countries from the 1940s to the 1970s. Organochlorine pesticides were banned in the late 1970s because of their persistence in the environment and their carcinogenic and mutagenic effects. Organochlorine pesticides were employed in huge amounts in the Spanish archipelago of the Canary Islands; the authors, therefore, evaluated the role played by organochlorine pesticides exposure on bladder cancer. Serum levels of the most prevalent organochlorine pesticides used in the agriculture of these Islands (dichlorodiphenyltrichloroethane [p,p'-DDT], and its metabolites dichlorodiphenyldichloroethylene [p,p'-DDE] and dichlorodiphenyldichloroethane [p,p'-DDD], hexachlorobenzene, hexachlorocyclohexane isomers, aldrin, dieldrin, endrin, heptachlor, cis-chlordane, trans-chlordane, α- and ß-endosulfan, endosulfan sulfate, methoxychlor, and mirex) were measured in 140 bladder cancer cases and 206 controls. GST-M1 and GST-T1 gene polymorphisms were genotyped by polymerase chain reaction (PCR)-based methods. These results showed that serum levels of organochlorine pesticides did not increase bladder cancer risk. On the contrary, total burden of hexachlorocyclohexanes was found to be negatively associated to bladder cancer (odds ratio [OR] = 0.929, 95% confidence interval [CI]: 0.865-0.997; P = .041). This effect disappeared when the distribution of the gluthathione S-transferase polymorphisms was introduced in the statistical model. These results indicate that organochlorine pesticides are not a risk factor for bladder cancer. However, these findings provide additional evidence of gene-environment interactions for organochlorine pesticides and bladder cancer and reinforce the relevance of genes encoding xenobiotic-metabolizing enzymes in bladder cancer.

Mamografía con realce de contraste mediante técnica de energía dual / Dual-energy contrast-enhanced mammography

Es bien conocido que el grado de vascularización de las lesiones mamarias se relaciona con su malignidad. Por ello, las técnicas de diagnóstico por imagen que estudian funcionalmente las lesiones han cobrado mucha relevancia en los últimos años. La mamografía con medio de contraste y energía dual es una técnica de reciente aparición, aparentemente prometedora en el cáncer de mama, que informa del grado de vascularización de la lesión junto con la información morfológica habitual. El propósito de este artículo es presentar el estado actual de esta nueva técnica de imagen. Basándonos en una experiencia de 15 meses, ilustramos esta revisión con casos clínicos que nos permiten presentar también sus ventajas y limitaciones (AU)

The degree of vascularization in breast lesions is related to their malignancy. For this reason, functional diagnostic imaging techniques have become important in recent years. Dual-energy contrast-enhanced mammography is a new, apparently promising technique in breast cancer that provides information about the degree of vascularization of the lesion in addition to the morphological information provided by conventional mammography. This article describes the state of the art for dual-energy contrast-enhanced mammography. Based on 15 months’ clinical experience, we illustrate this review with clinical cases that allow us to discuss the advantages and limitations of this technique (AU)

Dual-energy contrast-enhanced mammography.

The degree of vascularization in breast lesions is related to their malignancy. For this reason, functional diagnostic imaging techniques have become important in recent years. Dual-energy contrast-enhanced mammography is a new, apparently promising technique in breast cancer that provides information about the degree of vascularization of the lesion in addition to the morphological information provided by conventional mammography. This article describes the state of the art for dual-energy contrast-enhanced mammography. Based on 15 months' clinical experience, we illustrate this review with clinical cases that allow us to discuss the advantages and limitations of this technique.

Potential adverse effects of inorganic pollutants on clinical parameters of loggerhead sea turtles (Caretta caretta): results from a nesting colony from Cape Verde, West Africa.

A large number of nesting loggerhead sea turtles (n = 201) were sampled to establish the blood levels of 11 elements (Cu, Mn, Pb, Zn, Cd, Ni, Cr, As, Al, Hg, and Se). Almost all of the samples showed detectable levels of these 11 elements, and Zn and Se exhibited the highest concentrations (median values as high as 6.05 and 2.28 µg/g, respectively). The median concentrations of the most toxic compounds, As, Cd, Pb, and Hg, were relatively low (0.38, 0.24, 0.06, and 0.03 µg/g, respectively). We also determined the haematological and biochemical parameters in a subsample of 50 turtles to evaluate the potential effects of these contaminants on clinical parameters and found several associations. Our study reinforces the usefulness of blood for the monitoring of the levels of contaminating elements and their adverse effects on blood parameters in sea turtles.

Pansteatitis associated with high levels of polychlorinated biphenyls in a wild loggerhead sea turtle Caretta caretta.

We describe the morphologic and toxicological findings in a case of pansteatitis in a stranded loggerhead sea turtle Caretta caretta. At necropsy, a large amount of adipose tissue in the celomic cavity showing very firm, yellow to orange irregular formations was observed. Histological lesions ranged from the infiltration of necrotic fat by scarce multinucleated giant cells and numerous macrophages containing xylene-insoluble lipopigment inclusions to the presence of several granulomas characterized by an irregular central necrotic area consisting of lipopigment surrounded by numerous multinucleated giant cells. Microbiological cultures were negative. Celomic fat was analyzed for PCBs and DDTs, resulting in very high levels of PCB 138, 153, 180 209 (3170, 2830, 980 and 1190 ng g-1, respectively). Although a nutritional cause cannot be ruled out, the high levels of PCBs detected in the celomic fat could have induced lipid peroxidation in adipocytes, resulting in cell damage, deposition of ceroid pigment and inflammatory response. This is the first report of pansteatitis in a wild sea turtle.

Polychlorobiphenyls and organochlorine pesticides in conventional and organic brands of milk: occurrence and dietary intake in the population of the Canary Islands (Spain).

The population of the Spanish archipelago of the Canary Islands has been studied in depth regarding its levels of contamination by organochlorine pesticides (OCs) and polychlorinated biphenyls (PCBs). Foodstuffs of animal origin, such as milk, are prominent contributors to the body burden of these contaminants. As this population presents one of the highest milk-intake in Spain and Europe, we evaluated the level of OCs and PCBs in 26 commercially available brands of milk (16 conventional and 10 organic brands) present in the market of these Islands, in order to estimate the relevance of milk as a source of these chemicals for the Canary population. Our findings showed that hexachlorobenzene, trans-chlordane, and PCB153 were present in almost all the samples with independence of the type of milk. For both types of milks, the concentration of OCs was very low, showing organic milks lower levels than conventional ones. As a consequence, the estimated daily intake for OCs was lower than the tolerable daily intake (TDI) established by International Agencies. The levels of PCBs in milk were also found to be very low, but, in this case, the situation was the opposite: there were higher levels of PCBs in organic than in conventional brands of milk. Unexpectedly, levels of dioxin-like PCBs (DL-PCBs) reached values higher than 25 pg WHO-TEQ g(-1) fat in percentile 75 for both types of milk indicating the existence of a number of brands of milk highly contaminated by these toxicants. Consequently, the population who consume the most contaminated milk brands could have estimated daily intakes well above the recommended TDI (2 pg WHO-TEQ kg(-1)b.w.d(-1)) established by European Union Authorities. These results are of concern if we consider the well known adverse health effects exerted by dioxin-like compounds.

Multigenerational study of the hepatic effects exerted by the consumption of nonylphenol- and 4-octylphenol-contaminated drinking water in Sprague-Dawley rats.

Our multigenerational study evaluates the hepatic effects of the xenoestrogens nonylphenol (NP), and 4-octylphenol (4OP) on male and female rats when they are exposed uninterruptedly, from conception to adult age, to tap water containing 25ppm of NP or 4OP. Our results showed that these compounds did not induce any change in liver/body weight ratio (relative liver weight, RLW). In the morphological analysis we did not find evident signs of cytotoxicity. The most relevant findings were the presence of both an increase in the apoptotic index and in the percentage of binuclear hepatocytes in livers from exposed animals. Additionally, our study revealed the presence of hepatocellular glycogenosis (mainly in 4OP-exposed rats): the type of glycogen accumulated was in aggregates (gamma-glycogen), a non-functional form of glycogen. This study demonstrates that, at levels close to those described in the environment, NP and 4OP are capable of inducing a number of hepatic effects, potentially related with adaptive, and/or metabolic alterations of liver tissue.

Increasing serum levels of non-DDT-derivative organochlorine pesticides in the younger population of the Canary Islands (Spain).

Organochlorine pesticides are a lipophilic class of chemicals that persist in the environment and tend to accumulate in human tissues for years. They came into widespread use in the late 1940s. Because of their capacity to bioaccumulate and biomagnify in food chains and their toxic effects, most of them were banned in industrialized countries, among them Spain, in the late 1970s and 1980s. In 1998 organochlorine pesticides were determined in a representative sample of a Spanish population (around 690 serum samples from people 6 to 75years old from the Canary Islands). Serum levels of lindane aldrin, dieldrin and endrin, were determined. Our results showed that a high percentage of samples presented detectable levels of some of the organochlorines measured, endrin being the most frequently detected (72%) and at highest concentration (mean 136.7ng/g fat). Mean concentrations of the main cyclodiene evaluated, dieldrin, was lower to those found in other Western populations. However, serum levels of lindane were higher than those described in North European populations. Influence of geographical and sociodemographic factors was evaluated. Urban populations showed the highest levels of dieldrin, while non-urban population showed the highest serum values of lindane, aldrin and endrin. Unexpectedly, serum values of lindane, aldrin and dieldrin were higher in younger than in older people. Subjects under 18years showed almost twice as high serum levels of lindane, aldrin and dieldrin than subjects of 65-75years. These results may well suggest that people living in the Canary Islands have been and are currently exposed to non-DDT-organochlorine pesticides. The type and source of exposure could vary between islands and type of habitat. Contaminated food and/or the environment could be related with this situation.

Photoaffinity labeling identification of a specific binding protein for the anabolic steroids stanozolol and danazol: an oligomeric protein regulated by age, pituitary hormones, and ethinyl estradiol.

We have demonstrated previously that both rat and human liver microsomes contain a highly specific binding protein for the anabolic steroids stanozolol (ST) and danazol (DA). In this study we solubilized the male rat liver ST-binding protein (STBP) and investigated the following parameters: 1) pharmacological properties, 2) hydrodynamic properties, 3) peptidic composition, 4) the effects of age and hypophysectomy, and 5) inducibility by 17alpha-ethinyl estradiol. We found that STBP is an integral protein bound to the endoplasmic reticulum. 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) provided its optimal solubilization without changes in its pharmacological properties, i.e. high specificity for ST and danazol, between natural steroids and ligands of low affinity glucocorticoid-binding sites or of progesterone-binding sites. Hydrodynamic properties of the STBP showed that it has a molecular mass of at least 118 kDa. SDS-PAGE of covalently labeled STBP under nonreducing conditions showed that [3H]ST binds to a 110-kDa protein. The STBP was resolved under reducing conditions into three peptides of 55, 31, and 22 kDa, respectively. STBP increased from immature to adult rats, and it dramatically decreased after hypophysectomy. Unlike the 22-kDa peptide, both the 55- and 31-kDa peptides drastically decreased in both immature and hypophysectomized rats. 17alpha-Ethinyl estradiol administration to immature or hypophysectomized rats induced the 55- and 31-kDa [3H]STBP to a greater extent than the 22-kDa peptide. Thus, STBP appears as an oligomeric protein composed of hormone-regulated peptides. The availability of solubilized STBP and the fact that it can be induced in vivo represent major steps toward the purification and functional significance of this unique steroid-binding protein.

Evaluation of acute and chronic hepatotoxic effects exerted by anabolic-androgenic steroid stanozolol in adult male rats.

Stanozolol (ST) is a 17alpha-alkyl anabolic-androgenic steroid (17alpha-AAS) often misused by athletes and bodybuilders. The use of anabolic-steroids by sportsmen and teenagers has increased dramatically, thus raising the question about their hepatotoxicity, specially those such as ST which are orally administered. Previously, we have reported diverse in vivo effects exerted by this steroid and published the existence of a highly specific ST-binding site in male rat liver microsomes. The existence of this binding site, the reported hepatic effects exerted in humans, and the very limited information about its potential hepatotoxicity led us to treat adult male rats acutely and chronically with ST and study different parameters that could indicate liver damage: serum levels of transaminases, concentration of monooxygenase enzymes in liver, liver membrane lipid peroxidation products, liver histopathology, and cell cycle/ploidy status of liver cells. In our study, no changes in serum transaminases or lipid peroxidation levels were obtained. However, acute stanozolol treatment significantly decreased the levels of cytochrome P450 (Cyt. P450) and cytochrome b5 (Cyt. b5) during the first 48 h of treatment, while subsequently, at 72 and 96 h, these microsomal enzymes underwent a significant increase in their levels. In sharp contrast with this response to acute treatment, the content of these two enzymes during chronic treatment showed an important decrease. Interestingly, acutely and chronically ST-treated livers showed slight to moderate inflammatory or degenerative lesions in centrilobular hepatocytes. Flow cytometric analysis demonstrated that both acute and chronic ST treatment were capable of increasing the percentage of S-phase fraction (%SPF) of liver cells. These findings taken together clearly show that this steroid is capable of altering the liver capacity for metabolizing xenobiotics and indicate that high doses of ST could exert a proliferative effect on liver cells. Such data should be considered in risk evaluations for this compound.

Identification of a specific binding site for the anabolic steroid stanozolol in male rat liver microsomes.

Male rat liver microsomes contain a [3H]dexamethasone binding site, capable of binding glucocorticoids and progesterone. We have shown previously that the 17 alpha-alkylated androgen, stanozolol, can inhibit the [3H]dexamethasone binding to microsomes through a negative allosteric mechanism, which gives rise to the possibility of its interaction with a different binding site. In this study, the existence of a single-saturating binding site, capable of binding the radioactive steroid with a maximum number of the specific binding site of 49 +/- 2 pmol/mg of protein and a Kd of 37 +/- 1.3 nM was demonstrated by using [3H]stanozolol. In competition experiments, only stanozolol and danazol were able to compete with [3H]stanozolol for its binding to microsomes, among more than 60 steroids and other compounds tested. The binding of [3H]stanozolol was depressed after protease treatment of the microsomes, or after the administration of cycloheximide to adult male rats for 24 hr, which suggest its proteic nature. The [3H]stanozolol binding site was detected in many tissues of the rat, with the highest concentrations being found in the liver. It was detected from birth, increasing afterward in concentration and reaching a peak at 2 to 3 months of age. This is the first experimental verification of the existence in liver microsomes of a specific binding site for some 17 alpha-alkylate androgens, such as stanozolol and danazol, different from the androgen receptor or the [3H]dexamethasone binding site.

[3H]dexamethasone binding activity in liver microsomes is modulated differently by 17 alpha-alkylated androgens and testosterone in vivo.

Rat liver microsomes contain a single class of steroid binding sites, capable of binding various glucocorticoids and progesterone. In a previous article, we have described the in vitro interaction of several androgens with this binding site. Unlike natural androgens, the 17 alpha-alkyl derivatives stanozolol and danazol were capable of interacting with this binding site through a negative allosteric pattern. Now, the effects these steroids exert on the microsomal [3H]dexamethasone binding site have been studied in vivo. The administration of a single dose of stanozolol to rats provoked a significant reduction in the microsomal [3H]dexamethasone binding capacity. This effect was maximal two hr after stanozolol administration and persisted for six hr. The restoration of the [3H]dexamethasone binding level after stanozolol administration was dependent on protein synthesis, since it was blocked by the concomitant administration of cycloheximide. None of the other androgens tested (danazol, methyltestosterone, fluoxymesterone, and testosterone propionate) was capable of provoking a similar effect when administered 2 or 24 hr prior to sacrifice. In rats treated for seven days with a daily dose of diverse androgens and sacrificed 24 hr after the last treatment, none of the 17 alpha-alkyl androgens assayed provoked significant changes in the microsomal [3H]dexamethasone binding level, although stanozolol, danazol, and methyltestosterone provoked a significant increase in glucocorticoid receptor concentration. In contrast, the administration of testosterone propionate provoked a 50% reduction in the [3H]dexamethasone binding level without causing changes in the glucocorticoid receptor concentration. These results provide new evidence on the existence of different effects on the liver of 17 alpha-alkyl androgens, compared to the effects produced by natural androgens.