Recombinant human erythropoietin prevents etoposide- and methotrexate-induced toxicity in kidney and liver tissues via the regulation of oxidative damage and genotoxicity in Wistar rats.

Etoposide (ETO) and methotrexate (MTX) are two effective chemotherapeutic drugs. However, the clinical use of these drugs is limited by its toxicity in normal tissues, especially in kidney and in liver tissues. Recombinant human erythropoietin (rhEPO), erythropoietin hormone, has also been shown to exert tissue protective effects. The purpose of this study was to explore the protective effect of rhEPO against oxidative stress and genotoxicity induced by ETO and MTX in vivo. Adult male Wistar rats were divided into 10 groups (6 animals each): control group, rhEPO alone group, ETO alone group, MTX alone group and rhEPO + ETO/MTX groups. In rhEPO + ETO/MTX groups, three doses of pretreatment with rhEPO were performed: 1000, 3000 and 6000 IU/kg. Our results showed that rhEPO pretreatment protects liver and kidney tissues against oxidative stress induced by the anticancer drugs. The glycoprotein decreased malondialdehyde (MDA) levels, reduced catalase activity and ameliorated glutathione depletion. Furthermore, we showed that rhEPO administration prevented drug-induced DNA damage accessed by comet test. Altogether, our results suggested a protective role of rhEPO, especially at 3000 IU/kg, against ETO- and MTX-induced oxidative stress and genotoxicity in vivo.

Complete necrosis of hepatocellular carcinoma after preoperative portal vein embolization: a case report.

BACKGROUND: Hepatocellular carcinoma has a poor prognosis; few patients can undergo surgical curative treatment according to Barcelona Clinic Liver Cancer guidelines. Progress in surgical techniques has led to operations for more patients outside these guidelines. Our case shows a patient with intermediate stage hepatocellular carcinoma presenting a good outcome after curative treatment. CASE PRESENTATION: We report the case of an 80-year-old Moroccan man, who was positive for hepatitis c virus, presenting an intermediate stage hepatocellular carcinoma (three lesions between 20 and 60 mm). He presented a complete tumor necrosis after portal vein embolization and achieved 24-month disease-free survival after surgery. CONCLUSIONS: Perioperative care in liver surgery and multidisciplinary discussion can help to extend indications for liver resection for hepatocellular carcinoma outside European Association for the Study of the Liver/American Association for the Study of Liver Diseases recommendations and offer a curative approach to selected patients with intermediate and advanced stage hepatocellular carcinoma.

Effect of recombinant human erythropoietin on mitomycin C-induced oxidative stress and genotoxicity in rat kidney and heart tissues.

Mitomycin C (MMC) is an antineoplastic agent used for the treatment of several human malignancies. Nevertheless, the prolonged use of the drug may result in a serious heart and kidney injuries. Recombinant human erythropoietin (rhEPO) has recently been shown to exert an important cytoprotective effect in experimental brain injury and ischemic acute renal failure. The aim of the present work is to investigate the cardioprotective and renoprotective effects of rhEPO against MMC-induced oxidative damage and genotoxicity. Our results showed that MMC induced oxidative stress and DNA damage. rhEPO administration in any treatment conditions decreased oxidative damage induced by MMC. It reduced malondialdehyde and protein carbonyl levels. rhEPO ameliorated reduced glutathione plus oxidized glutathione modulation and the increased catalase activity after MMC treatment. Furthermore, rhEPO restored DNA damage caused by MMC. We concluded that rhEPO administration especially in pretreatment condition protected rats against MMC-induced heart and renal oxidative stress and genotoxicity.

Zearalenone-induced changes in biochemical parameters, oxidative stress and apoptosis in cardiac tissue: Protective role of crocin.

Zearalenone (ZEN) is a mycotoxin from Fusarium species commonly found in food commodities and is known to cause reproductive disorders. Several in vivo studies have shown that ZEN is haematotoxic and hepatotoxic and causes several alterations of immunological parameters. Meantime, the available information on the cardiotoxic effects of ZEN is very much limited. In the present study, we investigated the toxic effects of ZEN in heart tissues of Balb/c mice. We demonstrated that ZEN (40 mg kg(-1) body weight (b.w.)) increased creatine phosphokinase, lactate dehydrogenase, aspartate transaminase, alanine transaminase, total cholesterol and triglyceride levels and induced oxidative stress as monitored by measuring the malondialdehyde level, the generation of protein carbonyls, the catalase and superoxide dismutase activity and the expression of the heat shock proteins (Hsp 70). We also demonstrated that acute administration of ZEN triggers apoptosis in cardiac tissue. Furthermore, we aimed to evaluate the safety and efficacy of crocin (CRO), a natural carotenoid, to prevent ZEN-induced cardiotoxicity in mice. In fact, combined treatment of ZEN with different doses of CRO (50, 100, and 250 mg kg(-1) b.w.) showed a significant reduction of ZEN-induced toxicity for all tested markers in a dose-dependent manner. It could be concluded that CRO was effective in the protection against ZEN-induced toxicity in cardiac tissue.

Tacrolimus and mycophenolate mofetil associations: Induction of oxidative stress or antioxidant effect?

Gastrointestinal risk factors after organ transplantation are prevalent, due to the chronic use of immunosuppressant. The immunosuppressive drugs such as tacrolimus/mycophenolate mofetil (TAC/MMF) association are the most commonly used therapy. TAC and MMF have been implicated in gastrotoxicity, but their direct effects, alone and combined, on intestinal cells are not completely elucidated. This study investigated the effect of TAC and MMF alone and combined on human colon carcinoma cells. Our results demonstrated that TAC and MMF individually inhibit clearly cells proliferation, enhanced free radicals, lipid peroxidation production, induced DNA lesions and reduced mitochondrial membrane potential. In this study, we also showed that the two molecules TAC and MMF combined at high concentrations amplified the cell damage. Furthermore, the TAC (5 µM) prevented cell death induced by MMF (half maximal inhibitory concentration (IC(50))). Also, MMF (50 µM) induced cytoprotection in HCT116 cells against TAC (IC(50)) toxicity. Our findings provide additional evidence that oxidative damage is the major contribution of TAC and MMF combined toxicities. In fact, MMF and TAC exert a gastroprotective effect by modulating reactive oxygen species production. These data underscore the pleiotropic effect of TAC and MMF on HCT116 cells that play a preventive and critical role on intestinal function.

Role of recombinant human erythropoietin against mitomycin C-induced cardiac, hepatic and renal dysfunction in Wistar rats.

Mitomycin C (MMC) is one of the most effective chemotherapeutic drugs. However, the dose of MMC is greatly limited by its toxicity in normal tissues. Recombinant human erythropoietin (rhEPO), an erythropoietic hormone, has also been shown to exert tissue protective effects. The purpose of this study was to explore the protective effect of rhEPO against MMC-induced heart, liver, and renal dysfunction. Adult male Wistar rats were divided into six groups (with six animals each), namely control, rhEPO alone group, MMC alone group, and rhEPO + MMC group (pre-, co-, and posttreatment conditions). The results showed that MMC induced a marked cardiac, renal, and liver failure characterized by a significant decrease in body weight, organs weight, and organs ratio and a significant increase in creatinine, blood urea nitrogen, alanine aminotransferase, aspartate aminotransferase, γ-glutamyl transferase, and conjugated and total bilirubin levels in serum. Histological examination showed that MMC caused liver alterations. rhEPO treatment restored body weight, organs weight, and organs ratio as well as serum biochemical parameters and histological damage caused by MMC exposure.

The protective effect of recombinant human erythropoietin against cisplatin-induced renal and hepatic dysfunctions in Wistar rats.

Cisplatin (Cisp) is one of the most effective chemotherapeutic drugs. However, the dose of Cisp is greatly limited by its toxicity. Recombinant human erythropoietin (rhEPO), a hormone that regulates hematopoiesis, has also been shown to exert tissue-protective effects. The purpose of this study was to explore the protective effect of rhEPO against Cisp-induced renal and liver dysfunctions. Adult male Wistar rats were divided into six groups of six each: control, rhEPO-alone group, Cisp-alone group and rhEPO + Cisp group (pretreatment, cotreatment and posttreatment conditions). Our results showed that Cisp-induced a marked renal and liver failure characterized by a significant decrease in body weight, organ weight and organ ratio and a significant increase in creatinine, blood urea nitrogen, alanine aminotransferase, aspartate aminotransferase, G-glutamyl transferase, alkaline phosphatase, bilirubin conjugated and bilirubin total levels in serum. Histological examination showed that Cisp caused kidney alterations. rhEPO treatments restored body weight, organ weight and organ ratio as well as serum biochemical parameters changed due to Cisp exposure.

Protective effect of aqueous extract of Allium sativum against zearalenone toxicity mediated by oxidative stress.

Zearalenone (ZEN) is a non-steroidal estrogenic mycotoxin produced by several species of Fusarium in cereals and agricultural products. It has been implicated in several mycotoxicosis in farm animals and in humans. Several reports suggest that oxidative damage seems to be a key determinant of ZEN induced toxicity in vitro and in vivo. The aim of the current study was to evaluate the protective effects of aqueous extract of Allium sativum (AEA), against ZEN-induced cytotoxicity, reactive oxygen species (ROS) generation and DNA fragmentation in cultured Vero cells. Indeed, cytotoxicity effects were studied using MTT viability assay, ROS generation measurement and catalase activity induction. To check whether the oxidative stress induction was associated to DNA lesions, we looked for DNA fragmentation using Comet test. Our results indicated that ZEN induced several toxic effects and significant alterations mediated by oxidative stress mechanism. Treatment by ZEN combined to the lowest dose of AEA (250 µg/ml) showed a significant reduction of ZEN induced damages for all tested markers and a noticeable reduction of DNA fragmentation. It could be concluded that A. sativum aqueous extracts is effective in the protection against ZEN hazards. This could be relevant, particularly with the emergent demand for natural products which may counteract the detrimental toxic effects mediated by oxidative stress process and therefore prevents multiple human diseases.

Leishmania (Viannia) braziliensis identification by PCR in the state of Para, Brazil.

The incidence of cutaneous leishmaniasis (CL) is increasing and there is limited surveillance of Leishmania species throughout the world. We identified the species associated with CL in a region of Amazonia, an area recognized for its Leishmania species variability. Clinical findings were analyzed and correlated with the species identified in 93 patients. PCR assays were based on small subunit ribosomal DNA (SSU-rDNA) and G6PD, and were performed in a laboratory located 3,500km away. Leishmania (V.) braziliensis was identified in 53 patients (57%). The other 40 patients (43%) carried a different species (including six cases of L. (L.) amazonensis). Molecular methods can be employed, using special media, to allow transport to distant laboratories. L. (V.) braziliensis is the most common species in the area of Para. The location of ulcers can suggest CL species.

Deoxynivalenol contamination in Tunisian barley in the 2009 harvest.

In Tunisia, barley is commonly used in human consumption in a variety of food forms. In this regard, a high quality of this agricultural product is always demanded by consumers. A survey of the natural occurrence of deoxynivalenol (DON), the most common Fusarium mycotoxin in small grain cereals, in barley harvested in the main cropping regions in Northern Tunisia in the 2009 harvest was conducted. A total of 72 samples were analysed for DON using high-performance liquid chromatography (HPLC) with a UV visible detector set at 220 nm. Between 36% and 100% of the samples were positive for DON with averages ranging from 1.2 to 2.4 mg kg(-1). A positive correlation between DON levels and temperature was seen; on the other side no correlation between DON contents and rainfall was observed. In this study we notably showed the effect of regions on DON contamination.

Cytotoxicity and related effects of T-2 toxin on cultured Vero cells.

T-2 toxin belongs to a group of mycotoxins synthesized by Fusarium fungi that are widely encountered as natural contaminants of certain important agricultural commodities particularly, cereals. Upon exposure, T-2 toxin causes severe human and animal diseases. It is considered to be a major causative agent in fatal alimentary toxic aleukia (ATA) in humans. In this study, cytotoxicity and apotosis induction by T-2 toxin was investigated in vitro on Vero cell line using the MTT and the neutral red viability assays, the induction of lipid peroxidation, the decrease of macromolecule levels (protein, DNA and RNA), DNA fragmentation and caspase-3-dependent apoptosis induction. Our results showed that T-2 toxin reduced cell viability correlated to an impairment of macromolecule levels. It also increased MDA formation, induced DNA fragmentation showed by DNA laddering patterns on agarose gel electrophoresis. This fragmentation is in relation with apoptosis induction which was confirmed by activation of caspase-3, and depletion of the mitochondrial membrane potential reflecting a mitochondrial dysfunction.

Zearalenone induces chromosome aberrations in mouse bone marrow: preventive effect of 17beta-estradiol, progesterone and Vitamin E.

The cytogenetic effect of zearalenone (ZEN), a non-steroidal estrogenic mycotoxin, was evaluated in vivo, in mouse bone marrow cells, by assessing the percentage of cells bearing different chromosome aberrations. The studies included different conditions for animal treatment, as follows: (1) single intraperitoneal (ip) injection, (2) repeated ip injections, (3) pre-treatment for 24h with Vitamin E (Vit E), and (4) pre-treatment for 4h with 17beta-estradiol (17beta-Est) or progesterone (Prog). ZEN induced different types of chromosome aberrations, which was concentration-dependent (2-20 mg/kg bw). These doses corresponded to 0.4-4% of the LD50 in the mouse. Interestingly, when the dose of ZEN (40 mg/kg) was fractionated into four equivalent doses (4 x 10 mg/kg bw), into three doses (15 + 10 + 15 mg/kg bw), or into two equivalent doses (2 x 20 mg/kg bw), given every 24 h, the percentage of chromosome aberrations increased significantly. This finding suggests that ZEN proceeds by reversible binding on receptors that could become saturated, and that it damages the chromosomes in a 'hit and go' manner. Furthermore, pre-treatment of animals with 17beta-estradiol or progesterone significantly decreased the percentage of chromosome aberrations, suggesting that (i) these hormones bind to the same cytoplasmic receptors transported into the nucleus to elicit DNA damage, (ii) they may play a role in preventing chromosome aberrations induced by ZEN. Similarly, Vit E prevented these chromosome aberrations indicating that Vit E, previously reported to prevent most of the toxic effects induced by ZEN, may also bind to the same receptors.

Cytotoxicity, inhibition of DNA and protein syntheses and oxidative damage in cultured cells exposed to zearalenone.

Mycotoxins are toxic metabolites of various fungi commonly found in feed and foodstuff and can cause very serious health problems in animals as well as in humans. Zearalenone (ZEN), a mycotoxin produced by various Fusarium species has several adverse effects. Indeed, ZEN has strong estrogenic activity associated with hyperestrogenism and several physiological alterations of the reproductive tract. Moreover, ZEN was shown to be hepatotoxic, haematotoxic, immunotoxic and genotoxic. The exact mechanism of ZEN toxicity is not completely established. The observed strong estrogenic effect of ZEN resulting from its competition with 17beta-estradiol in the binding to estrogen receptors is generally considered to underline most toxic effects of ZEN, but estrogenic activity alone cannot explain the diverse and apparent adverse effects. The objective of the present study was to determine the involvement of other possible mechanisms in ZEN induced toxicity. Cytotoxicity, cell cycle perturbation, inhibition of protein and DNA synthesis as well as the presumed later marker of oxidative stress, malondialdehyde, were monitored in Vero and Caco-2 cells exposed to ZEN. Our results showed that ZEN reduces cell viability correlated to cell cycle perturbation, inhibits protein and DNA syntheses and increases MDA formation in both cell lines in concentration-dependant manner. We assumed that cytotoxicity and oxidative damage are additional mechanisms of ZEN mediated toxicity.

Karyomegaly of tubular cells as early stage marker of the nephrotoxicity induced by ochratoxin A in rats.

Cases of karyomegaly were described by Sclare and by Mihatch in patients affected with tubular-interstitial nephropathy. The Karyomegalic cells showed enlarged nuclei with accumulation of genetic material. No aetiology was suggested. Our study of rats experimentally intoxicated by ochratoxin A, a well-known nephrotoxic compound, indicates the presence of karyomegaly with alteration of the tubular tissue. In control animals no karyomegalic cells were detected. These observations suggest that karyomegaly with megacytosis may be caused by the nephrotoxic ochratoxin A in the kidney. In addition abnormal mitosis together with karyomegalic cells were observed at an earlier stage of the intoxication (30 days) suggesting possible regeneration if the OTA insults are stopped. After 90 days of treatment, the degeneration increased and only karyomegalic and apoptotic-like cells were observed indicating that the regeneration no longer occurs and that the degeneration becomes irreversible.

[Concurrent leishmaniasis and human immunodeficiency virus (HIV) infection: a study of four cases]. / Concomitância de leishmanioses e infecção pelo vírus da imunodeficiência humana (HIV): estudo de quatro casos.

Few cases of concurrent leishmaniasis and HIV infection have been reported in Brazil, despite both infections being in expansion. Two cases of visceral leishmaniasis and two cases of mucocutaneous leishmaniasis are discussed. Disseminated skin and oral lesions were found in the patients with the cutaneous form of the disease. Prolonged fever, hepatosplenomegaly and pancytopenia were the main manifestations of the visceral form. The CD4 T lymphocyte count was low in all cases. Direct examination of bone marrow aspirate for leishmania and biopsy of cutaneous lesions are the techniques of choice to confirm diagnosis. Pentavalent antimonials and amphotericin B are preferred drugs for the treatment of leishmaniasis, including patients with AIDS. The authors recommend the inclusion of this parasitosis in the differential diagnosis of opportunistic diseases in patients with AIDS.

Induction of a SOS repair system in lysogenic bacteria by zearalenone and its prevention by vitamin E.

Zearalenone (Zen) is an oestrogenic mycotoxin produced by several Fusarium species in cereals. It induces modifications of haematological parameters in rats with cytotoxicity and inhibition of macromolecular synthesis (nucleic acids and protein). Zen and its metabolites have oestrogenic and anabolic activities and interact with human oestrogen receptors. Zen and its metabolites showed a positive DNA damaging effect in recombination tests with Bacillus subtilis. It induces sister chromatid exchange and chromosomal aberration in CHO cells. Zen was found to be capable of inducing DNA-adduct formation in mouse liver. The genotoxicity of Zen was questionable until the last decade when increasing data tended to show this toxin to be genotoxic in vivo. However the mechanism of its genotoxicity and mutagenicity has not been completely clarified. The present investigations were designed to show whether Zen induces an SOS-DNA repair response in lysogenic bacteria which have an integrated lambda-bacteriophage in their genome. Zen was found to be genotoxic in the bacterial systems from a concentration of 1.50 mM and it was also bactericidal (IC50 = 1.45 mM). In addition vitamin E (6.0-12.0 mM) added 1 h prior to the toxin proved to prevent both the genotoxic and bactericidal effects of Zen. This vitamin could be active both as an antioxidant and as a radical scavenger. The specificity of this prevention is probably due to the similarity of structure between vitamin E and Zen.

Retinol, ascorbic acid and alpha-tocopherol prevent DNA adduct formation in mice treated with the mycotoxins ochratoxin A and zearalenone.

Ochratoxin A (OTA), and zearalenone (ZEN), two mycotoxins, have been implicated in numerous mycotoxicoses in farm animals and are genotoxic. Several adducts were detected in mouse and rat kidney after a single administration of OTA and in mice organs after zearalenone treatment which induces hepatocellular adenomas. The effects of some vitamins such as retinol (A), ascorbic acid (C) and alpha-tocopherol (E), which are known to act as superoxide anion scavengers, were tested on OTA genotoxicity. Pretreatment of mice by vitamin E decreased DNA adducts by 80% in kidney. Vitamin A decreased DNA adduct levels by 70% and Vitamin C by 90% in kidney. In the same way, pretreatment of female mice with alpha-tocopherol before administration of zearalenone inhibited significantly DNA adduct formation in liver and in kidney. The total DNA adduct level after E treatment was decreased by 45% and 58% in liver and kidney respectively.

Paracoccidioidomycosis in a boy infected with HIV.

Paracoccidioidomycosis has been rarely reported among HIV patients, despite being an endemic mycosis in Latin America. The present report illustrates a case of PM in a teenager infected with HIV. Clinicians must be aware that this mycosis may occur in young HIV infected patients who live in endemic areas.

Zearalenone induces modifications of haematological and biochemical parameters in rats.

Zearalenone produced by the fungus Fusarium roseum causes important perturbations in the gestation cycle of the rat with hormonal disorders and infertility. In order to find out other eventual toxic effects, female rats were given intraperitoneally (i.p.) (1.5, 3 and 5 mg/kg) zearalenone in sterile olive oil. Forty-eight hours later, some blood parameters changed (hematocrit, MCV, the number of platelets and WBC) as well as some biochemical markers such as aminotransferases (ALT, AST), alkaline phosphatase (ALP), serum creatinine, bilirubin, indicating liver toxicity, and likely impairment of blood coagulation process.

Genotoxicity of zearalenone, an estrogenic mycotoxin: DNA adduct formation in female mouse tissues.

Zearalenone is a non-steroidal estrogenic mycotoxin produced by several species of Fusarium which colonize maize, barley, oats, wheat and sorghum and have been implicated in numerous mycotoxicoses in farm animals, especially pigs. In a NTP mouse study a dose-related incidence of hepatocellular adenomas was seen in female mice. A limited number of genotoxicity assays have been conducted with zearalenone. Zearalenone was found to be negative in the Salmonella typhimurium assay. It was also negative in a eukaryotic cell mutation assay with Saccharomyces cerevisae. However, zearalenone and its estrogenic metabolites showed a positive DNA damaging effect in recombination tests with Bacillus subtilis. In this study DNA adducts in female mice and rat treated i.p or orally with zearalenone were determined using a 32P-postlabeling method. Several DNA adducts (12-15) were found in the kidney and liver of female mice treated with a single dose of zearalenone (2 mg/kg i.p. or orally). The total DNA adduct levels reached 114 +/- 37 and 1393 +/- 324 adducts/10(9) nucleotides respectively in kidney and liver after i.p. treatment and 548 +/- 50 adducts/10(9) nucleotides in liver after oral treatment. In mice ovary DNA adducts appeared only after repeated doses (1 mg/kg body wt on days 1, 5, 7, 9 and 10). The total DNA adducts after 10 days in this organ were 17 +/- 5 adducts/10(9) nucleotides. Some adducts were common to all organs. Others were specific to an organ. In contrast, no DNA adducts could be detected in rat organs after i.p. treatment. These results confirm the genotoxicity of zearalenone and its ability to induce hepatocellular adenomas, rather than tumours of genital organs, in mice.