Listeria monocytogenes is an important foodborne pathogen, markedly persistent even in harsh environments and responsible for high hospitalization and mortality rates. The aim of the present study was to detect the strains circulating in Sicily over a five-year period and characterize their antimicrobial resistance profiles. The key element of this study was the sharing of data among various entities involved in food control and clinical surveillance of listeriosis in order to develop an integrated approach for this pathogen. A total of 128 isolates were analyzed, including 87 food-source strains and 41 clinical specimens. Whole-genome sequencing (WGS) was performed for sequence type (ST) and clonal complex (CC) identification through multilocus sequence typing (MLST) analysis. Antimicrobial resistance was assessed using the Kirby–Bauer method. The majority of strains belonged to serotype IVb (34/41 and 53/87 of clinical and food-source isolates, respectively) and were subtyped as CC2-ST2 (28/34 and 41/53 of clinical and food-source isolates respectively). Most of the isolates were susceptible to the main antimicrobials recommended for treatment of listeriosis. Resistance (R) and intermediate resistance (I) percentages worthy of attention were found against oxacillin (R: 85.9%) and clindamycin (I: 34.6%) in the food-source isolates and trimethoprim/sulfamethoxazole (R: 29.23%) in the clinical isolates. Also, 7.7% of the food-source isolates were multidrug resistant. Our results highlight how the punctual comparison between food and clinical strains is an essential tool for effectively tracking and preventing foodborne outbreaks.
Bivalves can concentrate biological and chemical pollutants, causing foodborne outbreaks whose occurrence is increasing, due to climatic and anthropic factors that are difficult to reverse, hence the need for improved surveillance. This study aimed to evaluate the hygienic qualities of bivalves sampled along the production and distribution chain in Sicily and collect useful data for consumer safety. Bacteriological and molecular analyses were performed on 254 samples of bivalves for the detection of enteropathogenic Vibrio, Arcobacter spp., Aeromonas spp., Salmonella spp., and beta-glucuronidase-positive Escherichia coli. A total of 96 out of 254 samples, collected in the production areas, were processed for algal biotoxins and heavy metals detection. Bacterial and algal contaminations were also assessed for 21 samples of water from aquaculture implants. Vibrio spp., Arcobacter spp., Aeromonas hydrophila, Salmonella spp., and Escherichia coli were detected in 106/254, 79/254, 12/254, 16/254, and 95/254 molluscs, respectively. A total of 10/96 bivalves tested positive for algal biotoxins, and metals were under the legal limit. V. alginolyticus, A. butzleri, and E. coli were detected in 5, 3, and 3 water samples, respectively. Alexandrium minutum, Dinophysis acuminata, Lingulodinium polyedra, and Pseudonitzschia spp. were detected in water samples collected with the biotoxin-containing molluscs. Traces of yessotoxins were detected in molluscs from water samples containing the corresponding producing algae. Despite the strict regulation by the European Commission over shellfish supply chain monitoring, our analyses highlighted the need for efficiency improvement.
Notification of foodborne outbreaks has been mandatory in Europe since 2005, and surveillance is carried out along the entire food chain. Here we report the results obtained from laboratory investigations about four cases of foodborne outbreaks that occurred in Sicily between 2009 and 2016, deemed to be related to staphylococcal enterotoxins (SEs) and coagulase-positive Staphylococci (CPS) by the Local Public Health Authority. Primosale cheese samples were processed by culture methods for enumeration of CPS and immunoenzymatic assays for detection and differentiation of the SEs possibly contained in food samples. In all cases, the mistrusted foods were found to be contaminated by CPS at bacterial loads between 5 and 8 log CFU/g and contained SE type C (SEC). The reported data confirm the risk of staphylococcal food poisoning associated with the consumption of raw milk cheese. The SEC is the most commonly occurring SE in goat milk and dairy products and the most represented enterotoxin in Sicilian dairy products. Our results highlighted the need for improving the current monitoring efficiency and implementing the available laboratory methods to collect more faithful epidemiological data on the current prevalence of staphylococcal toxins in the food chain, including SEs currently not detectable by validated analytical methods.
Antioxidants are the most popular active ingredients in anti-aging cosmetics as they can restore the physiological radical balance and counteract the photoaging process. Instead of adding pure compounds into the formulations, some “precious” vegetable oils could be used due to their content of tocopherols, phenols, vitamins, etc., constituting a powerful antioxidant unsaponifiable fraction. Here, electron paramagnetic resonance (EPR) spectroscopy coupled with spin trapping was proven to provide a valid method for evaluating the antioxidant properties and the oxidative resistance of vegetable oils which, following UV irradiation, produce highly reactive radical species although hardly detectable. Extra virgin olive oil, sweet almond oil, apricot kernel oil, and jojoba oil were then evaluated by using N-t-butyl-α-phenylnitrone as a spin trapper and testing different UV irradiation times followed by incubation for 5 to 180 min at 70 °C. The EPR spectra were manipulated to obtain quantitative information useful for comparing the different tested samples. As a result, the knowledge acquired via the EPR analyses demonstrated jojoba oil as the best of the four considered oils in terms of both starting antioxidant ability and oxidative stability overtime. The obtained results confirmed the usefulness of the EPR spin trapping technique for the main proposed purpose.
Seafood can vehiculate foodborne illnesses from water to humans. Climate changes, increasing water contamination and coastlines anthropization, favor the global spread of Vibrio spp. and the occurrence of antibiotic-resistant isolates. The aim of this study was to evaluate the spread of potentially pathogenic Vibrio spp. in fishery products collected in Sicily and to assess their antibiotic resistance. Bacteriological and molecular methods were applied to 603 seafood samples to detect V. parahaemolyticus, V. cholerae, V. vulnificus, and Vibrio alginolyticus in order to assess their pathogenicity and antimicrobial resistance. About 30% of bivalves and 20% of other fishery products were contaminated by Vibrio spp.; V. parahaemolyticus accounted for 43/165 isolates, 3 of which were carrying either tdh or trh; V. cholerae accounted for 12/165 isolates, all of them non-O1 non-O139 and none carrying virulence genes; and V. vulnificus accounted for 5/165 isolates. The highest rates of resistance were observed for ampicillin, but we also detected strains resistant to antibiotics currently included among the most efficient against Vibrio spp. In spite of their current low incidence, their rise might pose further issues in treating infections; hence, these results stress the need for a continuous monitoring of antimicrobial resistance among fishery products and an effective risk assessment.
The spread of multidrug resistant (MDR) Salmonella strains, along the poultry supply chain, can represent a relevant threat to human health. This study aimed to evaluate the prevalence and antimicrobial resistance of Salmonella spp. isolated from poultry meat for human consumption. Between 2019 and 2021, 145 samples were analyzed according to ISO 6579-1:2017. The strains isolated were identified by using biochemical-enzymatic assays and serotyping, according to the Kauffmann-White-Le Minor scheme. The antibiotic susceptibility tests were determined using the Kirby-Bauer method. Forty Salmonella spp. strains were isolated and serotyping showed Salmonella Infantis to be predominant. 80% of the isolated strains were MDR and identified as S. Infantis. This study confirms the circulation of MDR Salmonella isolated from poultry meat and highlights the predominance of the S. Infantis serovar, which represents an emerging risk factor under the One Health holistic approach.
This preliminary study aimed to detect biological and chemical contaminants in vegetables sold in Sicily for human consumption, assess the spread of antimicrobial-resistant (AMR) strains in these foods, and characterize their antimicrobial-resistance genes. A total of 29 fresh and ready-to-eat samples were analyzed. Microbiological analyses were performed for the detection of Salmonella spp. and the enumeration of Enterococci, Enterobacteriaceae, and Escherichia coli. Antimicrobial resistance was assessed by the Kirby-Bauer method, according to the Clinical and Laboratory Standards Institute guidelines. Pesticides were detected by high-performance liquid chromatography and gas chromatography coupled with mass spectrometry. No samples were contaminated by Salmonella spp., E. coli was detected in 1 sample of fresh lettuce at a low bacterial count (2 log cfu/g). 17.24% of vegetables were contaminated by Enterococci and 65.5% by Enterobacteriaceae (bacterial counts between 1.56 log cfu/g and 5.93 log cfu/g and between 1.6 log cfu/g and 5.48 log cfu/g respectively). From 86.2% of vegetables, 53 AMR strains were isolated, and 10/53 isolates were multidrug resistant. Molecular analysis showed that the blaTEM gene was detected in 12/38 β-lactam-resistant/intermediate-resistant isolates. Genes conferring tetracycline resistance (tetA, tetB, tetC, tetD, tetW) were detected in 7/10 isolates. The qnrS gene was detected in 1/5 quinolone-resistant isolates, the sulI gene was detected in 1/4 sulfonamide- resistant/intermediate-resistant isolates and the sulIII gene was never detected. Pesticides were detected in 27.3% of samples, all of which were leafy vegetables. Despite the satisfactory hygienic status of samples, the high percentage of AMR bacteria detected stresses the need for an effective monitoring of these foods as well as adequate strategies to counteract the spread of AMR bacteria along the agricultural chain. Also, the chemical contamination of vegetables should not be underestimated, especially considering that leafy vegetables are commonly consumed raw and that no official guidelines about maximum residue limits of pesticides in ready-to-eat vegetables are available.
Salmonella spp. are among the most frequent causes of foodborne diseases, and the increasing occurrence of MDR strains is an additional cause for concern. In the three-year period 2019–2021, we collected Salmonella spp. strains isolated from different food categories analysed in the context of Regulation (EC) No 2073/2005 in order to assess their antibiotic susceptibility profiles and ESBL production. To determine the susceptibility profiles and identify MDR strains, we used the Kirby–Bauer method to test 17 antibiotics. Double-disc and PCR testing then allowed us to assess the production of ESBLs and the presence of beta-lactamase resistance genes. Phenotypic tests showed that 36 out of 67 strains were MDR and 52.7% of these were ESBL producers. Finally, molecular investigations conducted on ESBL-producing strains revealed the presence of blaSHV, blaCTX-M and blaTEM genes. Our results confirmed the prevalence of S. Infantis, an MDR strain and ESBL producer, in chicken meat. This suggests that further research on the prevalence of antibiotic resistance genes (ARGs) in foodborne strains is needed, especially from a One Health perspective.
The traditional cultural method (PCR and Real-Time PCR) for Salmonella spp. detection and identification is laborious and time-consuming. A qualitative LAMP method detecting Salmonella spp. was validated in compliance with ISO 16140:2016. The results show a relative accuracy, sensitivity, and specificity of 100% in comparison with the reference method ISO 6579-1:2017; the LOD50 was set as 0.4 CFU/g. Additionally, a field study was carried out comparing the LAMP kit, a commercially available Real-Time PCR kit (FoodProof Salmonella, Biotecon Diagnostics), and the reference cultural method. The Salmonella spp. LAMP kit was suitable for reliable detection of Salmonella spp., simplifying and reducing the extent and the steps of the analytical process. A total of 105 samples of raw poultry meat were screened for the presence of Salmonella spp. according to three methods: the LAMP kit Salmonella spp. (Enbiotech), the Real-Time PCR kit FoodProof Salmonella (Biotecon), and the reference cultural method. Using these three methods, only one sample out of the 105 (0.95%) tested was positive for Salmonella spp. This sample was further investigated using the reference method described in ISO 6579-3:2014, in order to characterise the Salmonella strain. Following this further biochemical identification and serological typing, the isolate was characterised as Salmonella Infantis.
The aim of the study was to evaluate the occurrence of Arcobacter spp. in food samples collected from Sicilia region. A total of 91 food products of animal origin (41 meat, 17 fresh milk, 18 shellfish) and 15 samples of fresh vegetables, were examined by cultural method and confirmed by biochemical analysis and PCR methods. The detection of Arcobacter spp. was performed, after selective enrichment, on two selective agar plates: Arcobacter agar and mCCD (modified charcoal cefoperazone deoxycholate) agar supplemented with CAT (Cefoperazone, Amphotericin B and Teicoplanin). Arcobacter species were isolated using the membrane filtration technique. In 13 (14.3%) out of the 91 tested samples, the presence of Arcobacter spp. was found: the isolates were confirmed by multiplex PCR and identified as belonging to the species A. butzleri and A. cryaerophilus. The highest prevalence rate was observed in chicken meat (8.8%) followed by shellfish (3.3%). Negative results have been obtained for raw milks and vegetables samples. The preliminary study highlights the importance of this emerging pathogen and the need for further studies on its prevalence and distribution in different types of food for human consumption.
Extra-intestinal E. coli are emerging as a global threat due to their diffusion as opportunistic pathogens and, above all, to their wide set of antibiotic resistance determinants. There are still many gaps in our knowledge of their origin and spread pathways, although food animals have been adjudicated vehicles for passing mult-drug resistant bacteria to humans. This study analyzed 46 samples of meat purchased from retail stores in Palermo in order to obtain quinolone-resistant E. coli isolates. Strains were screened for their phylogenetic groups, ST131-associated single nucleotide polymorphisms (SNPs), and then typed by ERIC-PCR. Their set of virulence factors, namely, kpsMII, papA, sfaS, focG, iutA, papC, hlyD, and afa genes, were investigated and their fluoroquinolone-resistance determinants evaluated. The data obtained show a dramatically high prevalence of multidrug resistance patterns in the Palermo area, with 28% of the isolates having virulence factor genes typical of ExPEC strains. No B2 group or ST131 strains were detected. Moreover, 20% of our isolates showed positivity to all the plasmid-mediated quinolone resistance (PMQR) determinants, showing a potential to transfer these genes among other bacteria. Therefore, these data underline the possibility that food animals and, specifically, poultry in particular may be a significant source of resistant bacterial strains, posing a potential zoonotic risk.
Fresh produce occupies an increasingly important place in the human food supply because of its health-promoting nutritional properties. Most fresh produce is eaten raw or after minimal processing and, consequently, pathogen contamination can represent a serious health risk. There has been an increase in foodborne outbreaks and cases associated with fresh produce, but literature data about the prevalence of pathogen contamination are inconsistent. This study was undertaken to assess the hygienic quality and the prevalence of the most common bacterial pathogens in fresh produce sold in retail markets in Sicily. A total of 125 samples of different types of vegetables were examined by standardized microbiological methods.
A case of staphylococcal food poisoning was observed in two individuals of the same family after consumption of primosale, a semiripened sheep cheese produced in Sicily. Staphylococcus aureus isolated from the cheese produced enterotoxin C (SEC) and carried both the enterotoxin C (sec) and the toxic shock syndrome toxin (tsst-1) gene. Following this case, an extensive survey was conducted on 971 food samples (raw milk, cheese, meat, and food preparations). S. aureus was detected in 102 of 971 food samples, from all types of food with the exception of ricotta cheese. The tsst-1 gene was present in 42% of the strains, either alone or in combination with other toxin genes. The enterotoxin C gene was the most represented enterotoxin, but it was only found in dairy products. Six S. aureus isolates carried the sea gene alone, two isolates carried both sea and seb, and one isolate carried both sea and sec. A significant percentage (46%) of all isolates carried a toxin gene, creating significant concern that virulent S. aureus can be transmitted through food in Sicily.
Annals of the New York Academy of SciencesVolume 784, Issue 1 p. 70-84 Multiple Estrogen Function in Human Prostate Cancer Cellsa GIUSEPPE CARRUBA, GIUSEPPE CARRUBA Hormone Biochemistry Laboratories School of Medicine University of Palermo Palermo, ItalySearch for more papers by this authorMARIA D. MICELI, MARIA D. MICELI Hormone Biochemistry Laboratories School of Medicine University of Palermo Palermo, ItalySearch for more papers by this authorLOREDANA COMITO, LOREDANA COMITO Experimental Oncology and Molecular Endocrinology Units Palermo Branch of the National Cancer Institute of Genoa “M. Ascoli”. Cancer Hospital Center Palermo, ItalySearch for more papers by this authorROSARIA FARRUGGIO, ROSARIA FARRUGGIO Hormone Biochemistry Laboratories School of Medicine University of Palermo Palermo, ItalySearch for more papers by this authorCARMELA M. G. SORCI, CARMELA M. G. SORCI Hormone Biochemistry Laboratories School of Medicine University of Palermo Palermo, ItalySearch for more papers by this authorGIUSEPPA OLIVERI, GIUSEPPA OLIVERI Hormone Biochemistry Laboratories School of Medicine University of Palermo Palermo, ItalySearch for more papers by this authorROSALBA AMODIO, ROSALBA AMODIO Hormone Biochemistry Laboratories School of Medicine University of Palermo Palermo, ItalySearch for more papers by this authorMARCO DI FALCO, MARCO DI FALCO Hormone Biochemistry Laboratories School of Medicine University of Palermo Palermo, ItalySearch for more papers by this authorDOMENICO D'AMICO, DOMENICO D'AMICO Hormone Biochemistry Laboratories School of Medicine University of Palermo Palermo, ItalySearch for more papers by this authorLUIGI A. M. CASTAGNETTA, LUIGI A. M. CASTAGNETTA Hormone Biochemistry Laboratories School of Medicine University of Palermo Palermo, Italy Experimental Oncology and Molecular Endocrinology Units Palermo Branch of the National Cancer Institute of Genoa “M. Ascoli”. Cancer Hospital Center Palermo, ItalySearch for more papers by this author GIUSEPPE CARRUBA, GIUSEPPE CARRUBA Hormone Biochemistry Laboratories School of Medicine University of Palermo Palermo, ItalySearch for more papers by this authorMARIA D. MICELI, MARIA D. MICELI Hormone Biochemistry Laboratories School of Medicine University of Palermo Palermo, ItalySearch for more papers by this authorLOREDANA COMITO, LOREDANA COMITO Experimental Oncology and Molecular Endocrinology Units Palermo Branch of the National Cancer Institute of Genoa “M. Ascoli”. Cancer Hospital Center Palermo, ItalySearch for more papers by this authorROSARIA FARRUGGIO, ROSARIA FARRUGGIO Hormone Biochemistry Laboratories School of Medicine University of Palermo Palermo, ItalySearch for more papers by this authorCARMELA M. G. SORCI, CARMELA M. G. SORCI Hormone Biochemistry Laboratories School of Medicine University of Palermo Palermo, ItalySearch for more papers by this authorGIUSEPPA OLIVERI, GIUSEPPA OLIVERI Hormone Biochemistry Laboratories School of Medicine University of Palermo Palermo, ItalySearch for more papers by this authorROSALBA AMODIO, ROSALBA AMODIO Hormone Biochemistry Laboratories School of Medicine University of Palermo Palermo, ItalySearch for more papers by this authorMARCO DI FALCO, MARCO DI FALCO Hormone Biochemistry Laboratories School of Medicine University of Palermo Palermo, ItalySearch for more papers by this authorDOMENICO D'AMICO, DOMENICO D'AMICO Hormone Biochemistry Laboratories School of Medicine University of Palermo Palermo, ItalySearch for more papers by this authorLUIGI A. M. CASTAGNETTA, LUIGI A. M. CASTAGNETTA Hormone Biochemistry Laboratories School of Medicine University of Palermo Palermo, Italy Experimental Oncology and Molecular Endocrinology Units Palermo Branch of the National Cancer Institute of Genoa “M. Ascoli”. Cancer Hospital Center Palermo, ItalySearch for more papers by this author First published: April 1996 https://doi.org/10.1111/j.1749-6632.1996.tb16229.xCitations: 19 a This study was partially funded by the Italian Association for Cancer Research (AIRC) and by the National Research Council (CNR) Special Project “Aging” (Grant No. 95.01017.PF40). R. Farruggio and L. Comito hold grants from, respectively, the National Cancer Institute (IST) of Genoa and the Italian Association GROCTA. AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Citing Literature Volume784, Issue1Basis for Cancer ManagementApril 1996Pages 70-84 RelatedInformation
This paper summarizes our most recent results of steroid enzyme studies on cultured breast and endometrial cancer cells. It deals mainly with estrogen 17 beta-hydroxysteroid oxidoreductase (17 beta HSOR) activity, which presides over estradiol (E2) and estrone (E1) interconversion, a major metabolic pathway of estrogens. Assessment of either the oxidative or reductive component of 17 beta HSOR was carried out on intact cells by means of an original approach based on reverse phase-high performance liquid chromatography and radioactive detection on line. This system allows the continuous monitoring of both precursor degradation and formation of several radiometabolites to assess rates and direction of steroid metabolism. Overall, hormone-responsive, estrogen receptor (ER)-positive cells, regardless of whether they were derived from breast (MCF7) or endometrial (Ishikawa) tumor tissues, showed a prevalence for reductive metabolism (E1-->E2), whilst oxidative pathways (E2-->E1) were largely dominant in non-responsive, ER-poor mammary (MDA-MB231) and endometrial (HEC-1A) cells. The above estimates of 17 beta HSOR activity were at variance with those obtained using the classical enzymology approach, not only in quantitative terms (being markedly lower using intact cell analysis), but also because the prevalent direction of estrogen metabolism was often reversed. Although striking methodological differences may well account for this discrepancy, intact cell analysis is undoubtedly more similar to the in vivo state than the artificial requirements of classical enzymology procedures.
We report that growth of LNCaP human prostate cancer cells is significantly stimulated (up to 120% above control) by physiological estradiol (E(2)) concentrations. This growth increase appears to be comparable to that induced by either testosterone or dihydrotestosterone, as also reported by others. This paper presents novel illustrative evidence for estrogen-binding proteins and messenger RNA transcripts in LNCaP cells. In fact, 1) the reverse transcriptase-polymerase chain reaction system documented normal messenger RNA for estrogen receptors (ER); 2) the radioligand binding assay allowed the detection of high affinity, reduced capacity binding sites in both soluble and nuclear cell fractions; and 3) the immunocytochemical analysis showed a consistently intensive staining for both ER and progesterone receptors. Compared to other human estrogen-responsive mammary cancer cells, MCF7 and ZR75-1, ER expression in LNCaP cells was not significantly lower, as shown by levels of the ER transcripts, number of sites per cell, or femtomoles per mg DNA as well as the percentage and intensity of immunocytochemical staining. A relative estimate of ER expression obtained by matching LNCaP with another human prostate cancer cell line, PC3, always displayed significantly and consistently higher levels in LNCaP cells. The detection of relatively high type I ER content in either cell compartment of LNCaP cells was paralleled by a highly intensive staining for progesterone receptors. In addition, evidence that the synthetic androgen R1881 did not compete for type I binding of E(2) and that any E(2)-induced growth was completely reversed by the pure antiestrogen ICI-182,780, but unaffected by the antiandrogen Casodex, clearly suggests that the biological response of LNCaP cells to E(2) is mediated via its own receptor.
In order to measure the formation and degradation rates of estradiol by human breast cancer cells, after assessing the biochemical basis of hormone responsiveness and growth response to estrogens, we considered both responsive, estrogen receptor (ER) positive, and non-responsive, ER-negative, breast cancer cell lines, i.e. MCF7, ZR75-1 and MDA-MB231. To this end, we employed a novel "intact cell" approach which allows us, after 24 h incubation, to analyze several enzyme activities in sequence, concurrently with the monitoring of labeled precursor degradation. Our investigations led to the following evidence: (a) the reductive activity of the 17 beta-hydroxysteroid oxoreductase (17 beta-HSOR) appears to be higher than the oxidative only in responsive, ER-rich MCF7 and ZR75-1 cells, as also previously observed by others; (b) this activity is, on the contrary, much lower in MDA-MB231 cells and other unresponsive, ER-poor breast cancer cell lines; (c) conversely, the oxidative activity shows an opposite pattern, being limited in MCF7 and ZR75-1 cells and much higher in MDA-MB231 cells. Overall, a 17 beta-HSOR reductive pathway prevails in both MCF7 and ZR75-1 cells, whilst the oxidative pathway is prevalent in MDA-MB231 cells, leading to a large formation of estrone that is no further metabolized, at least in the experimental conditions used. Our results may provide a likely explanation of previous data on the different estrogen content of breast tumor tissues.
In this paper we report that two human long-term endometrial cancer cell lines, Ishikawa and HEC-1A, exhibit quite different abilities in metabolizing estrogens. As a matter of fact, incubation of Ishikawa cells with close-to-physiological concentrations of estradiol (E(2)) as precursor resulted in: (1) elevated formation (up to 90%) of E(2)-sulphate (E(2)-S), using lower precursor concentrations; (2) very limited conversion to estrone (E(1)) (<10% at 24 h incubation), as either free or sulphate; and (3) low but consistent production of other estrogen derivatives, such as 2-hydroxy-estrogens and estriol. Conversely, scant amounts (if any) of E(2)-S were found in HEC-1A cells, while no detectable formation of other estrogen metabolites could be observed after 24h. On the other hand, E(1) production was significantly greater (nearly 60% at 24 h) than in Ishikawa cells, a large proportion of E(1) (over 50% of the total) being formed after only 6 h incubation using time-course experiments. The hypothesis that E(2) metabolism could be minor in Ishikawa cells as a consequence of the high rate of E(2)-S formation encountered is contradicted by the evidence that conversion to E(1) also remains limited in the presence of much lower E(2)-S amounts, seen using higher molar concentrations of precursor. Overall, we observe that 17 beta-hydroxysteroid dehydrogenase (17 beta-HSD) activity diverges significantly in intact Ishikawa and HEC-1A endometrial cancer cells. This difference could not merely be accounted for by the diverse amounts of substrate (E(2)) available to the cells, nor may it be imputed to different levels of endogenous estrogens. It should rather be sought in different mechanisms controlling 17 beta-HSD activity or, alternatively, in the presence of distinct isoenzymes in the two different cell types.