Renal myxoma: an unexpected differential diagnosis.

Myxomas are rare mesenchymal tumors that can appear in many anatomical locations, although they are mainly seen in heart and skin. To date, only twelve cases of pure renal myxomas have been reported in the literature. We describe a case of a young Cuban woman with an asymptomatic irregular cyst lesion in her left kidney which was eventually diagnosed as renal myxoma. We also provide radiological and pathological studies.

Are fish and standardized FETAX assays protective enough for amphibians? A case study on Xenopus laevis larvae assay with biologically active substances present in livestock wastes.

Biologically active substances could reach the aquatic compartment when livestock wastes are considered for recycling. Recently, the standardized FETAX assay has been questioned, and some researchers have considered that the risk assessment performed on fish could not be protective enough to cover amphibians. In the present study a Xenopus laevis acute assay was developed in order to compare the sensitivity of larvae relative to fish or FETAX assays; veterinary medicines (ivermectin, oxytetracycline, tetracycline, sulfamethoxazole, and trimethoprim) and essential metals (zinc, copper, manganese, and selenium) that may be found in livestock wastes were used for the larvae exposure. Lethal (LC(50)) and sublethal effects were estimated. Available data in both, fish and FETAX studies, were in general more protective than values found out in the current study, but not in all cases. Moreover, the presence of nonlethal effects, caused by ivermectin, zinc, and copper, suggested that several physiological mechanisms could be affected. Thus, this kind of effects should be deeply investigated. The results obtained in the present study could expand the information about micropollutants from livestock wastes on amphibians.

Identity, virulence genes, and clonal relatedness of Aeromonas isolates from patients with diarrhea and drinking water.

Among 800 stool specimens from patients with diarrhea submitted by Primary Care Centers for routine analysis to the Hospital of León (NW Spain) Microbiology and Parasitology Service, 32 (4%) were tested positive for Aeromonas spp. Mixed infections with other enteric pathogens occurred in 12 patients. A. caviae was isolated from 23 clinical specimens. There were also patients infected with A. media, A. hydrophila, A. bestiarum, and A. veronii biovar veronii. All but three isolates carried one or more of the virulence genes. The incidence of the alt, hlyA, aerA, ast, and laf genes was 71.9, 28.1, 25.0, 18.8, and 9.4%, respectively. The alt(+)/ast(+) combination was detected in four isolates and the aerA(+)/hlyA(+) combination was detected in the two A. hydrophila isolates. None of the strains harbored the TTSS, stx1, or stx2 genes and nine bore plasmids. Thirty clinical isolates and a collection of 12 A. caviae and A. media strains obtained from León municipal drinking water over the study period were typed by pulsed-field gel electrophoresis (PFGE). PFGE patterns revealed genetic relatedness and persistence over time among water isolates and some clinical isolates. Interestingly, one A. caviae (aerA(-)/hlyA(-)/alt(+)/ast(-)/laf(+)) human isolate and two A. caviae (aerA(-)/hlyA(-)/alt(+)/ast(-)/laf(+)) drinking water isolates had indistinguishable PFGE patterns, suggesting waterborne infection.

Effect assessment of reclaimed water and carbamazepine exposure on the thyroid axis of X. laevis: Apical and histological effects.

There is increasing environmental concern about the constant presence of pharmaceuticals and personal care products (PPCPs) in surface water, generally attributed to water discharge from wastewater treatment plants (WWTPs) that are unable to completely remove these compounds. The slight, but continuous, presence of these contaminants in reclaimed water (RW) poses a risk of chronic and sublethal toxicity, and the thyroid axis can likely be a target of many of these PPCPs. In this work, we addressed the effects of RW on the Xenopus laevis thyroid system. The Amphibian Metamorphosis Assay (AMA test) was used with modifications by exposing X. laevis tadpoles to RW samples, and to RW spiked with carbamazepine (CBZ) at 100 and 1000 higher than the average levels environmentally relevant (RW 100× and RW 1000×, respectively). Carbamazepine was selected because it is considered a marker of anthropogenic pollution and could have a potential effect on the thyroid axis. The morphological endpoints and histological alterations to the thyroid gland were evaluated. The results suggested the stimulation of the thyroid gland from exposures to the RW samples, supported by tadpoles' accelerated development and by the histological alterations observed in the thyroid gland. Developmental acceleration was also seen in the tadpoles exposed to the RW-100× and -1000× samples at comparable levels to those seen in exposures to RW samples alone. Hence CBZ did not seem to increase the effects of RW on the thyroid axis. Overall, our results suggested endocrine effects of these RW samples regardless of the CBZ concentration.

Sublethal and chronic effects of reclaimed water on aquatic organisms. Looking for relationships between physico-chemical characterisation and toxic effects.

The use of reclaimed water for irrigation and aquaculture purposes is generally considered a reliable alternative for sustainable water management in regions with water scarcity. Many organic compounds, generally called compounds of emerging concern (CECs), have been detected in reclaimed water, which implies continuous exposure for aquatic organisms. To date no quality criteria have been proposed for this group of compounds. This work aims to assess the acute, sublethal and chronic effects of reclaimed water using two representative organisms of the aquatic compartment; the green alga Chlorella vulgaris and the microcrustacean Daphnia magna. The study comprises the 72 h-algal growth inhibition test, the D. magna feeding bioassay and the D. magna reproduction test. The results highlighted, for the selected characterised compounds, no differences in the concentrations between the different tertiary WWTP treatments, except for the particular case of carbamazepine. Considering seasonality, no differences were observed between the two different sample collection campaigns. The sublethal and chronic effects observed for these samples could not be explained by the lower concentrations found in the chemical characterisation. However, in the majority of cases, dilution of raw reclaimed water reduced the toxic effects of these samples. Several interactions among compounds can affect the mixture's toxicity. Canonical correlation analyses (CCA) were included to explore the potential relationships between the physico-chemical characterisation of reclaimed water and effects on aquatic organisms. The results corroborated the toxic effect of some pharmaceuticals, in particular beta-blockers and antibiotics, on the growth and yield of green algae, as well as inhibition of daphnia reproduction. Thus the CCA methods could help to elucidate the potential relationships between the physico-chemical characterisation and toxic effects by considering all the potential interactions.

Cryopreservation of captive roe deer (Capreolus capreolus) semen.

To address the need to preserve current genetic diversity before it is lost forever; further studies to adapt assisted reproductive technologies to various endangered species are needed, among other things. Roe deer (Capreolus capreolus), an over abundant wild deer, can serve as model species to develop or improve sperm cryopreservation of threatened or endangered deer species. The aim of this study was to compare the ability of three diluents (Berliner Cryomedium [BC]; Tris, citric acid, glucose [TCG]; TES, Tris, glucose) to support chilling, cryopreservation (with 5% glycerol; G) and postthaw incubation (at 22 °C and 37 °C) of roe deer spermatozoa collected by electroejaculation. Berliner Cryomedium was the diluent that better preserved roe deer spermatozoa during refrigeration, able to maintain motility for at least 14 days, longer than the other extenders. BC + G was the extender of choice for cryopreservation, showing higher viability compared with TCG + G (66.7 ± 3.4 vs. 54.5 ± 6.5; P < 0.05) and higher level of acrosome integrity compared with TES, Tris, glucose + G (79.4 ± 3.4 vs. 67.9 ± 5.0; P < 0.05). Maintaining the samples at 22 °C after thawing presented higher values in various parameters compared with 37 °C. The knowledge gained through this study can potentially act as a preliminary step toward development of new protocols to help increase the reproductive success of biologically similar, yet endangered, wild species.

Lipopolysaccharide-induced DNA damage is greatly reduced in rats treated with the pineal hormone melatonin.

The ability of melatonin to influence lipopolysaccharide (LPS)-induced genotoxicity was tested using micronuclei as an index in both bone marrow and peripheral blood cells of rats. LPS was given as a single dose of 10 mg/kg. Melatonin (5 mg/kg) was injected prior to LPS administration and thereafter at 6 h intervals to the conclusion of the study (72 h). The number of micronucleated polychromatic erythrocytes increased significantly after LPS administration both in cells from peripheral blood and bone marrow. Melatonin administration to LPS-treated rats highly significantly reduced micronuclei formation in both peripheral blood and bone marrow cells beginning at 24 h after LPS administration and continuing to the end of the study. In blood the increase in micronuclei formation was time-dependent in LPS-treated rats with peak values being reached at 36-48 h. The ability of melatonin to reduce LPS-related genotoxicity is likely related to its antioxidant activity.

Postoperative contamination after using vancomycin and gentamicin during phacoemulsification.

OBJECTIVE: To investigate if the addition of vancomycin and gentamicin to the irrigating solutions during phacoemulsification with lens implantation reduces the incidence of positive postoperative intraocular cultures. DESIGN: Two-part double-masked, placebo-controlled, randomized clinical trial. PATIENTS: In the preliminary study, intracameral antibiotic concentrations were measured immediately after surgery (in 10 eyes) and 2 hours after surgery (in 10 eyes) in patients treated with antibiotics. In the primary study, 120 eyes underwent uncomplicated surgery. The treatment group and the placebo group were composed of 60 eyes each. INTERVENTION: The treatment group received vancomycin, 20 micrograms/mL, and gentamicin, 8 micrograms/mL, in the irrigating fluid. The placebo group received only irrigating fluid. All patients in the primary study underwent anterior chamber aspiration following surgery, and culturing was performed 2 hours later. MAIN OUTCOME MEASURE: Identification and quantification of positive cultures in thioglycolate broth and chocolate agar. RESULTS: In the preliminary study, the half-life of both intraocular antibiotics was less than 2 hours. In the primary study, intraocular aspirates yielded positive cultures in 3 specimens (5.0%) in the antibiotic-treated group and in 7 specimens (12.0%) in the placebo group. CONCLUSIONS: Although we found a higher rate of positive postoperative cultures in the placebo group (odds ratio = 2.51), 2 hours of contact between the antibiotic solution and bacteria did not produce results that reached statistical significance (P =.18) to support adding vancomycin and gentamicin to the irrigating solutions during phacoemulsification. Further studies are needed to evaluate the clinical implications of using antibiotics in irrigating solutions.

Nocturnal decreases in nitric oxide and cyclic GMP contents in the chick brain and their prevention by light.

The diurnal variations in the contents of nitric oxide (NO) and cyclic GMP were studied in the chick brain. NO and cyclic GMP contents in the chick brain were lower at night than during the day and were inversely correlated with high night-time tissue melatonin levels. Furthermore, when animals were kept in light at night, tissue melatonin levels remained at low diurnal values, whereas NO and cyclic GMP contents remained high. Since we have previously shown that physiological concentrations of melatonin inhibit nitric oxide synthase (NOS) activity in different brain areas, the nocturnal decrease in brain NO and cyclic GMP contents may be, in part, a consequence of the nocturnal inhibitory effect of melatonin on NOS activity.

Melatonin stimulates brain glutathione peroxidase activity.

Exogenously administered melatonin causes a 2-fold rise in glutathione peroxidase activity within 30 min in the brain of the rat. Furthermore, brain glutathione peroxidase activity is higher at night than during the day and is correlated with high night-time tissue melatonin levels. Glutathione peroxidase is thought to be the principal enzyme eliminating peroxides in the brain. This antioxidative enzyme reduces the formation of hydroxyl radicals formed via iron-catalyzed Fenton-type reactions from hydrogen peroxide by reducing this oxidant to water. Since the hydroxyl radical is the most noxious oxygen radical known, induction of brain glutathione peroxidase might be an important mechanism by which melatonin exerts its potent neuroprotective effects.

Rhythms of glutathione peroxidase and glutathione reductase in brain of chick and their inhibition by light.

Melatonin was recently shown to be a component of the antioxidative defense system of organisms due to its free radical scavenging and antioxidant activities. Pharmacologically, melatonin stimulates the activity of the peroxide detoxifying enzyme glutathione peroxidase in rat brain and in several tissues of chicks. In this report, we studied the endogenous rhythm of two antioxidant enzymes, glutathione peroxidase and glutathione reductase, in five regions (hippocampus, hypothalamus, striatum, cortex and cerebellum) of chick brain and correlated them with physiological blood melatonin concentrations. Glutathione peroxidase exhibited a marked 24 h rhythm with peak activity in each brain region which had acrophases about 8 h after lights off and about 4 h after the serum melatonin peak was detected. Glutathione reductase activity exhibited similar robust rhythms with the peaks occurring roughly 2 h after those of glutathione peroxidase. We suggest that neural glutathione peroxidase increases due to the rise of nocturnal melatonin levels while glutathione reductase activity rises slightly later possibly due to an increase of its substrate, oxidized glutathione. The exposure of chicks to constant light for 6 days eliminated the melatonin rhythm as well as the peaks in both glutathione peroxidase and glutathione reductase activities. These findings suggest that the melatonin rhythm may be related to the nighttime increases in the enzyme activities, although other explanations cannot be excluded.

Acutely administered melatonin reduces oxidative damage in lung and brain induced by hyperbaric oxygen.

Hyperbaric oxygen exposure rapidly induces lipid peroxidation and cellular damage in a variety of organs. In this study, we demonstrate that the exposure of rats to 4 atmospheres of 100% oxygen for 90 min is associated with increased levels of lipid peroxidation products [malonaldehyde (MDA) and 4-hydroxyalkenals (4-HDA)] and with changes in the activities of two antioxidative enzymes [glutathione peroxidase (GPX) and glutathione reductase (GR)], as well as in the glutathione status in the lungs and in the brain. Products of lipid peroxidation increased after hyperbaric hyperoxia, both GPX and GR activities were decreased, and levels of total glutathione (reduced+oxidized) and glutathione disulfide (oxidized glutathione) increased in both lung and brain areas (cerebral cortex, hippocampus, hypothalamus, striatum, and cerebellum) but not in liver. When animals were injected with melatonin (10 mg/kg) immediately before the 90-min hyperbaric oxygen exposure, all measurements of oxidative damage were prevented and were similar to those in untreated control animals. Melatonin's actions may be related to a variety of mechanisms, some of which remain to be identified, including its ability to directly scavenge free radicals and its induction of antioxidative enzymes via specific melatonin receptors.

Influence of lithium salts on chick pineal gland melatonin secretion.

Pineal melatonin secretion exhibits a rhythm which is identified as a circadian cycle with a period of between 20 and 28 h in most animals. The melatonin rhythm has an acrophase of about -30 degrees. The aim of this study was determine the effect of lithium on the melatonin rhythm since lithium is usually used as a therapeutic agent in manic-depressive disease. Lithium induced a delay of the melatonin acrophase which reached a value of about -45 degrees; lithium also produced another peak which had an acrophase at about +60 degrees. We speculate that lithium influences either inositol phosphate turnover or it merely changes the profile of the melatonin rhythm.

Effect of calcium on melatonin secretion in chick pineal gland I.

Melatonin is the neurohormone which is synthesized by the pineal gland and secreted rhythmically. The role of calcium in the activation of melatonin production remains unknown. In this study, we demonstrated that calcium input participates in the regulation of chick pineal gland. Pineal glands from Gallus domesticus were perifuse with Krebs medium (controls) or with Krebs medium plus drugs (ethylene glycol tetraacetic acid (EGTA) or calcium ionophore A23187). When EGTA was added to the perifusion medium, free extracellular calcium concentrations were dramatically decreased and melatonin synthesis was decreased. On the other hand, when the calcium ionophore A23187 was added to the perifusion medium, chick pineal glands exhibited a marked increase in secretion of melatonin. No effects were observed when chick pineal glands were treated with drugs during or after the time of the natural peak levels. We propose that calcium input from extracellular medium and output from intracellular calcium reserves are primary mechanisms in the activation of melatonin synthesis in the chick pineal gland.

Effects of ethylene glycol tetraacetic acid, A23187 and calmodulin, calcium activated neutral proteinase antagonists on melatonin secretion in perifused chick pineal gland.

We have recently described, using perifused pineal glands, that calcium influx participates in the activation of chick pineal gland. This study shows that the loss of perifused chick pineal gland activity is a complex process which seems to involve the release of calcium from intracellular stores, calmodulin and calcium-activated neutral protease (CANP). Pineal glands were perifused with Krebs medium (controls) or with Krebs medium plus the drugs ethylene glycol tetraacetic acid (EGTA; calcium chelator), A23187 (calcium ionophore), EGTA plus A23187 (extra-intra cellular calcium chelation), trifluoperazine and CGS9343B (calmodulin inhibitors), and E-64 (CANP inhibitor) at the time of the natural peak of melatonin release. When EGTA or A23187 were added to the perifusion medium, no effects were observed. On the other hand, when the calcium chelator EGTA plus A23187 (free extra and intracellular calcium levels were dramatically decreased), trifluoperazine, CGS 9343B or E-64 were added to the perifusion medium melatonin synthesis increased significantly and was sustained for 8 h. We propose a prominent role for calcium output from intracellular stores in regulating melatonin production primarily by acting on Ca-calmodulin and calcium-activated neutral protease.

Effect of iron and estrogen on melatonin secretion by the chicken pineal gland.

Estrogen effects on pineal secretion of melatonin are controversial. Some feel that estrogen inhibits melatonin output in vitro but not in vivo. Melatonin levels vary with the age in chickens where circulating estrogen levels also vary. Laying hens have minimal melatonin levels and maximal serum iron concentrations. Thus, we reasoned that iron released by estrogen may inhibit melatonin secretion from the chick pineal gland. The present study shows that perifusion of estrogen-treated chick pineal glands with several concentrations of iron greatly inhibited melatonin secretion.

Melatonin reduces paraquat-induced genotoxicity in mice.

The protection afforded by melatonin against paraquat-induced genotoxicity in both bone marrow and peripheral blood cells of mice was tested using micronuclei as an index of induced chromosomal damage. Melatonin (2 mg/kg) or an equal volume of saline was injected i.p. into mice 30 min prior to the i.p. administration of paraquat (two injections of 15 mg/kg; the paraquat injections were given with a 24 h interval) and thereafter at 6 h intervals to the conclusion of the study (72 h). Using fluorescence microscopy, the number of micronuclei in polychromatic erythrocytes (MN-PCE) per 2000 PCE (1000 PCE/slide) per mouse was counted both in blood and bone marrow, and the ratio of PCE to normochromatic erythrocytes (NCE) (PCE/NCE) was calculated. Paraquat treatment increased the number of MN-PCE at 24, 48, and 72 h, both in peripheral blood and bone marrow cells, while no differences were observed in the PCE/NCE ratio. Melatonin inhibited the paraquat-induced increase in MN-PCE by more than 50% at 48 and 72h. Paraquat toxicity is believed to be due to free radical generation. Since melatonin is known to be an efficient free radical scavenger, it is concluded that melatonin's protection against paraquat-induced genotoxicity is mediated, at least in part, by its free radical scavenging activity.

Genotoxicity of paraquat: micronuclei induced in bone marrow and peripheral blood are inhibited by melatonin.

The ability of melatonin to influence paraquat-induced genotoxicity was tested using micronucleated polychromatic erythrocytes as an index of damage in both bone marrow and peripheral blood cells of mice. Melatonin (10 mg/kg) or an equal volume of saline were administered intraperitoneally (ip) to mice 30 min prior to an ip injection of paraquat (20 mg/kgx2), and thereafter at 6-h intervals until the conclusion of the study (72 h). The number of the micronucleated polychromatic erythrocytes increased after paraquat administration both in peripheral blood and bone marrow cells. Melatonin administration to paraquat-treated mice significantly reduced micronuclei formation in both peripheral blood and bone marrow cells; these differences were apparent at 24, 48 and 72 h after paraquat administration. The induction of micronuclei was time-dependent with peak values occurring at 24 and 48 h. The reduction in paraquat-related genotoxicity by melatonin is likely due in part to the antioxidant activity of the indole. We did not observe effects of melatonin over paraquat in paraquat+melatonin groups incubated at 0, 60 and 120 min. Mitomycin C, which was used as a positive control, also caused the expected large rises in micronuclei in both bone marrow and peripheral blood cells at 24, 48 and 72 h after its administration.

Value of limited necropsy in HIV-positive patients.

We performed limited autopsy with histological examination of tissue cores obtained percutaneously using the Tru-Cut needle and the Jamshidi trocar in 150 adult HIV-positive patients. Data were compared retrospectively with the antemortem clinical diagnosis. Eighty-one percent of the patients were male, and 78% were intravenous drug users. Specimens were obtained from the brain, liver, lung, bone marrow, and kidney of most patients. The main findings included liver cirrhosis in 22 cases (associated with HCV infection in 81%), Pneumocystis carinii pneumonia in 21, Cytomegalovirus (CMV) infection in 19, Mycobacterium avium-intracellulaire (MAI) infection in 17, bacterial pneumonia in 14, tuberculosis in 12, and lymphoma in 13 cases. Forty-six (30.6%) patients had at least one clinical diagnosis that was confirmed by autopsy, i.e., there was 40.6% agreement between pre- and postmortem findings. Forty-six (30.6%) patients had at least one clinical diagnosis that was not confirmed at autopsy, whereas 41 (27.3%) had at least one AIDS-related or unrelated disease that was not suspected clinically. The results obtained by limited autopsy are principally comparable to those achieved by full necropsy, with the advantages of decreasing the contagious risk, saving cost and time, including a rapid final diagnosis, and easily obtaining the consent for postmortem examination so that necropsy studies may be performed on a larger number of patients, thus contributing to a better understanding of the spectrum of HIV infection in our environment.

A system dynamic model for the assessment of different exposure routes in aquatic ecosystems.

With the exception of a limited number of highly persistent lipophilic pollutants, waterborne exposure is considered the most relevant exposure route for aquatic organisms, and even the only relevant exposure route in lower tier assessment, while the potential for bioaccumulation is only evaluated as secondary poisoning for birds and mammals. In addition, some evaluations consider that only the dissolved fraction is bioavailable for pelagic organisms. The lack of easy and cost-effective assessment methods is probably the main reason to disregard the contribution of other routes. This paper presents a system dynamics model that estimates the time-dependent accumulation of toxic chemicals through the food chain, and has also the potential to include the exposure from sediment. The generic ecosystem includes a primary producer and three levels of consumers, represented by unicellular algae, cladoceran invertebrates, cladoceran-eating fish and fish-eating fish, respectively. Nevertheless, the model, built using Microsoft Excel software, allows any number of levels and ecological-exposure relationships. The results obtained for four illustrative pollutants demonstrate that factors such as the binding to algae and other edible particles or food chain exposure for lipophilic non-persistent chemicals may constitute significant exposure routes. The current low tier European ecological risk assessment guidelines do not cover these aspects, and therefore may under estimate the real risk for both aquatic organisms and their predators. The model includes a simplified worst-case alternative that normally can be calculated using the existing information. Under a tiered approach, the simplified alternative is proposed to trigger the need of the toxicokinetics assays required to perform the complete time-dependent calculation.