Rat model of depending prostaglandin renal state: Effect of ketoprofen

Introduction .The renal prostaglandins (PGs), vasodilators, preserve kidney function during increased activity of the renin-angiotensin system or renal sympathetic nerves (renal PG-dependent state [RPGD]). Ketoprofen (Ket) inhibits cyclooxygenase and, therefore, the synthesis of PGs. The aim of this study was to determine, in the rat, the action of Ket in the renal histology and function in a RPGD state (stress of anesthesia and hemorrhage). Material and Methods . Twenty male Wistar rats, anesthetized with sodium pentobarbital, were randomly divided into two groups: G1-control ( n = 10) and G2-Ket ( n = 10) submitted to arterial hemorrhage of 30% of volemia (estimated as 6% of body weight) three times (10% each 10 min), 65 min after anesthesia. G2 animals received Ket, 1.5 mg. kg -1 , venously, 5 min after anesthesia and 60 min before the first hemorrhage moment (first moment of the study [M1]). Medium arterial pressure (MAP), rectal temperature (T), and heart rate were monitored. G1 and G2 received para-aminohippurate sodium (PAH) and iothalamate sodium (IOT) solutions during the entire experimental time in order to determine clearance of PAH (effective renal plasma flow [ERPF]) and clearance of IOT (glomerular filtration rate [GFR]) without urine collection (determination of blood concentrations of PAH and IOT through the high-performance liquid chromatography), filtration fraction (FF), and renal vascular resistance (RVR). The animals were sacrificed in M3, 30 min after the third hemorrhage (M2) moment, and the kidneys and blood collected during the hemorrhage periods were utilized for histological study and determinations of hematocrit (Ht), serum creatinine (S Cr ), ERPF, GFR, FF, and RVR, respectively. Results . There were significant reductions of MAP, T, and Ht and a significant increase of S Cr . During the experiment, ERPF and GFR did not change, but ERPF was always higher in G1 than in G2. Ket did not alter FF, which increased in G1 over the duration of experiment. The Ket group had significantly higher RVR than the control group. The histology verified that both G1 and G2 were similar for tubular dilation and necrosis, but they were significantly different for tubular degeneration: G1 > G2. Conclusion . The changes observed in kidney histology probably were determined by hemorrhage and hypotension. Ket inhibited the synthesis of PGs and diminished tubular degeneration.

Absence of carcinogenic and anticarcinogenic effects of annatto in the rat liver medium-term assay

Annatto (Bixa orellana L.) is a natural food colorant extensively used in many processed foods, especially dairy products. The lower cost of production and the low toxicity, make annatto a very attractive and convenient pigment in substitution to the many synthetic colorants. In the present study we investigate the carcinogenic and anticarcinogenic effects of dietary annatto in Wistar rat liver using the preneoplastic glutathione S-transferase (GST-P) foci and DNA damage biomarkers. Annatto, containing 5% bixin, was administered in the diet at concentrations of 20, 200, and 1000 ppm (0.07; 0.80 and 4.23 bixin/kg body wt/day, respectively), continuously during 2 weeks before, or 8 weeks after DEN treatment (200 mg/kg body wt, i.p.), to evaluate its effect on the liver-carcinogenesis medium-term bioassay. The comet assay was used to investigate the modifying potential of annatto on DEN (20 mg/kg body wt)-induced DNA damage. The results showed that annatto was neither genotoxic nor carcinogenic at the highest concentration tested (1000 ppm). No protective effects were also observed in both GST-P foci development and comet assays. In conclusion, in such experimental conditions, annatto shows no hepatocarcinogenic effect or modifying potential against DEN-induced DNA damage and preneoplastic foci in the rat liver. (C) 2004 Elsevier Ltd. All rights reserved.

Effect of Progesterone and Ionomycin on Domestic Cat Sperm Motility Patterns and Acrosome Reaction

This study was aimed at assessing the changes in sperm motion patterns and the percentage of acrosome reaction (AR) in domestic cat semen after treatment with either ionomycin or progesterone (P(4)). Ten ejaculates were collected from five tomcats using an artificial vagina, and were diluted, centrifuged and resuspended in a capacitation medium. Samples were evaluated and divided into seven equal aliquots and, after 2 h at 25 degrees C, were incubated for 30 min at 38 degrees C in 5% CO(2) and then analyzed. Computer-assisted sperm analysis and a combination of three fluorescent probes were used to assess sperm plasma, acrosomal membrane integrity and mitochondrial transmembrane potential. Thirty minutes after the start of incubation, P(4) was added (10 mu g/ml) to the P1 group. Groups P2 and P3 were supplemented with P(4) (10 and 20 mu g/ml, respectively) only after 2 h of incubation, and groups I1 and I2 were supplemented with ionomycin (4 and 8 mu M, respectively) 2 h after incubation. Group E was supplemented with ethanol (0.6%) at 2 h after incubation and group C received no supplementation. Ionomycin and P(4) treatments led to a hyperactivation-like sperm motion and an increase (p < 0.05) in the percentage of AR. Although a higher (p < 0.05) percentage of AR was obtained in group I2 when compared with all P(4) groups, a decrease (p < 0.05) in total and progressive motility was observed in I2 group. As I1 group was similar to I2 to induce AR without diminishing sperm motility, we can conclude that ionomycin at 4 mu M seems to be more suitable to trigger AR in domestic cat sperm.

Effect of melatonin on DNA damage of bovine cumulus cells during in vitro maturation (IVM) and on in vitro embryo development

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of prolonged erosive pH cycling on different restorative materials

This in vitro study evaluated the effect of a prolonged erosive pH cycling on the superficial microhardness change (SMHC) and the erosive wear of different restorative materials. Eighty enamel specimens with prepared cavities of 1.5 x 1.5 mm were randomly divided into eight groups according to the restorative materials used for the fillings (RMGI - resin-modified glass-ionomer, CGI - conventional glass-ionomer, CR- composite resin, A - amalgam) and immersion media used (ERO - erosive medium or SAL - artificial saliva). During 35 days, half of the specimens were immersed in a cola drink (ERO), for 5 min, three times a day, and they remained in SAL between the erosive cycles. The other half of the specimens was immersed in SAL only, for the entire experimental period (control). Data were tested for significant differences by anova and Tukey's tests (P < 0.05). Scanning electron microscopy images were made to illustrate the enamel erosive wear and restorative materials alterations. The mean SMHC (%) and mean erosive wear (mu m) of the materials were: RMGI-ERO (30/0.5); CGI-ERO (37/0.5); CR-ERO (-0.3/0.3); A-ERO (-4/0.3); RMGI-SAL (4/0.4); CGI-SAL (-6/0.4); CR-SAL (-3/0.2) and A-SAL (2/0.4). Scanning electron microscopy images showed pronounced enamel erosive wear on groups submitted to erosive pH cycling when compared with groups maintained in saliva. In conclusion, the prolonged pH cycling promoted significantly higher alterations (SMHC and erosive wear) on the glass-ionomer cements than the CR and amalgam.

In vitro effect of low-level laser on odontoblast-like cells

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of water storage and heat treatment on the cytotoxicity of soft liners

Objective: To evaluate the effect of water storage time on the cytotoxicity of soft liners.Methods: Sample discs of soft liners Dentusoft, Dentuflex, Trusoft, Ufi-Gel-P and denture base acrylic resin Lucitone-550 were prepared and divided into four groups: GN: No treatment, G24: Stored in water at 37 degrees C for 24 h; G48: Stored in water at 37 degrees C for 48 h, GHW: Immersed in water at 55 degrees C for 10 min. To analyse the cytotoxic effect, three samples of each group were placed in tubes with Dubelcco's Modified Eagle Mediums and incubated at 37 degrees C for 24 h. During this period, the toxic substances were leached to the culture medium. The cytotoxicity was analysed quantitatively by the incorporation of radioactivity H-3-thymidine checking the number of viable cells (synthesis of DNA). The data were statistically analysed using two-way ANOVA and Tukey's honestly significant difference tests (alpha = 0.05).Results: Treatments did not reduce the cytotoxicity effect of the soft liners (p > 0.05). It was found that Ufi-Gel-P had a non-cytotoxic effect, Trusoft had a slightly cytotoxic effect, Dentuflex had a moderated cytotoxic effect, Dentusoft alternated between slightly and non-cytotoxic effect, and Lucitone-550 had non-cytotoxic effect when stored in water for 48 h.Conclusion: The effect of water storage and the heat treatment did not reduce the cytotoxicity of the soft liners.

Effect of light curing sources on microhardness of different composite resins

This study evaluated the influence of light-curing units (LCUs) on Knoop microhardness (KHN) of different composite resins formulations. Four LCUs, one Quartz-Tungsten-Halogen (QTH) for 20 s, one Argon-Ion-Laser (AL) for 10 s, one Plasma-Arc-Curing (PAC) for 9 s, and one Light-Emitting-Diode (LED) for 20 s, and three composite resins, nanofill and easy cure (Filtek (TM) Supreme), microhybrid and medium cure (Herculite XRV), and microfill and difficult cure (Heliomolar) were used. Discs (4 x 2 mm(2)) of each composite resin were divided in 12 Groups and KHN was measured at the top (T) and bottom (B) surfaces. Data were analyzed using two-way ANOVA and Tukey's test (p < 0.05). Top presented significantly higher KHN than bottom surface for all composite resins and LCUs tested. Statistical significant differences were observed among the LCUs. At the bottom surface QTH and LED presented higher KHN than PAC and LA. However, at the top surface PAC and LA presented similar results than QTH for nanofill and microhybrid composite resins. Different LCUs play an important effect on Knoop microhardness and the composite resin formulations were significant factor on the photosensitivity.

Seasonal effect on Brazilian propolis antibacterial activity

The behavior of microorganisms towards the antibiotic action of propolis has been widely investigated. Since reports dealing with seasonal effect on propolis activity are not available, this assay was carried out aiming to observe the in vitro antimicrobial activity of propolis, collected during the four seasons, on bacterial strains isolated from human infections. Dilution of ethanolic extract of propolis (EEP) in agar was the method performed, with serial concentrations ranging from 0.4 to 14.0% (% v/v). The behavior of some bacteria was analysed according to the incubation period in medium plus propolis, and the survival curve was plotted. It was verified that the growth of Gram-positive bacteria is inhibited by low propolis concentrations (0.4%) whereas Gram-negative bacteria were less susceptible to this substance, the minimal inhibitory concentration ranging from 4.5 to 8.0%. There was no significant difference with regards to the seasonal effect on the survival curve of Staphylococcus aureus and Escherichia coli; after incubation with propolis, there was an efficient antimicrobial action, mainly towards Gram-positive bacteria. (C) 2000 Elsevier B.V. Ireland Ltd. All rights reserved.

Inhibitory effect of essential oils against Lactobacillus rhamnosus and starter culture in fermented milk during its shelf-life period

The use of essential oils in foods has attracted great interest, due to their antagonistic action against pathogenic microorganisms. However, this action is undesirable for probiotic foods, as products containing Lactobacillus rhamnosus. The aim of the present study was to measure the sensitivity profile of L. rhamnosus and a yogurt starter culture in fermented milk, upon addition of increasing concentrations of cinnamon, clove and mint essential oils. Essential oils were prepared by steam distillation, and chemically characterised by gas chromatography-mass spectrometry (GC-MS) and determination of density. Survival curves were obtained from counts of L. rhamnosus and the starter culture (alone and in combination), upon addition of 0.04% essential oils. In parallel, titratable acidity was monitored over 28 experimental days. Minimum inhibitory concentration values, obtained using the microdilution method in Brain Heart Infusion medium, were 0.025, 0.2 and 0.4% for cinnamon, clove and mint essential oils, respectively. Cinnamon essential oil had the highest antimicrobial activity, especially against the starter culture, interfering with lactic acid production. Although viable cell counts of L. rhamnosus were lower following treatment with all 3 essential oils, relative to controls, these results were not statistically significant; in addition, cell counts remained greater than the minimum count of 10(8)CFU/mL required for a product to be considered a probiotic. Thus, although use of cinnamon essential oil in yogurt makes starter culture fermentation unfeasible, it does not prevent the application of L. rhamnosus to probiotic fermented milk. Furthermore, clove and mint essential oil caused sublethal stress to L. rhamnosus.

Antiepileptic effect of acylpolyaminetoxin JSTX-3 on rat hippocampal CA1 neurons in vitro

The Joro spider toxin (JSTX-3), derived from Nephila clavata, has been found to block glutamate excitatory activity. Epilepsy has been studied in vitro, mostly on rat hippocampus, through brain slices techniques. The aim of this study is to verify the effect of the JSTX-3 on the epileptiform activity induced by magnesium-free medium in rat CA1 hippocampal neurons. Experiments were performed on hippocampus slices of control and pilocarpine-treated Wistar rats, prepared and maintained in vitro. Epileptiform activity was induced through omission of magnesium from the artificial cerebrospinal fluid (0-Mg2+ ACSF) superfusate and iontophoretic application of N-methyl-D-aspartate (NMDA). Intracellular recordings were obtained from CA] pyramidal neurons both of control and epileptic rats. Passive membrane properties were analyzed before and after perfusion with the 0-Mg2+ ACSF and the application of toxin JSTX-3. During the ictal-like activity, the toxin JSTX-3 was applied by pressure ejection, abolishing this activity. This effect was completely reversed during the washout period 2. when the slices were formerly perfused with artificial cerebrospinal fluid (ACSF) and again with 0-Mg2+ ACSF. Our results suggest that the toxin JSTX-3 is a potent blocker of induced epileptiform activity. (c) 2005 Elsevier B.V. All rights reserved.

Medium Chain Fatty Acids Are Selective Peroxisome Proliferator Activated Receptor (PPAR) gamma Activators and Pan-PPAR Partial Agonists

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of culture conditions on the biomass determination by ergosterol of Lentinus crinitus and Psilocybe castanella

Estimating fungal growth is important in processes of soil bioremediation. It has been demonstrated that ergosterol is a good indicator of fungal biomass in solid substrata. In the present study were evaluated the effects upon the ergosterol rate of Lentinus crinitus Berk. and Psilocybe castanella Peck through the culture conditions of these fungi, which are evaluated for the bioremediation of soils contaminated by organochlorates. A good correlation between fungal biomass and ergosterol was observed for both species. The culture conditions did not influence the ergosterol rate of L. crinitus. Yet the ergosterol rate of P. castanella was influenced from 35 days of culture and when grown in the presence of 15.00 g hexachlorobenzene l(-1) of culture medium. So it is possible to estimate growth of both species using ergosterol as indicator in processes of soil bioremediation since the influences observed in the ergosterol rate of P. castanella are considered.

The Use of Response Surface Methodology in Optimization of Lactic Acid Production: Focus on Medium Supplementation, Temperature and pH Control

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The effect of changes in the bulk dielectric constant on the DNA torsional properties

The effect of changes in the bulk dielectric constant on the DNA torsional properties was evaluated from plasmid circularization reactions. In these reactions, pUC18 previously linearized by EcoRI digestion was recircularized with T4 DNA ligase. The bulk dielectric constant of the reaction medium was decreased by the addition of different concentrations of neutral solutes: ethylene glycol, glycerol, sorbitol, and sucrose, or increased by the addition of glycine. The topoisomers generated by the ligase reaction were resolved by agarose-gel electrophoresis. The DNA twist energy parameter (K), which is an apparent torsional constant, was determined by linearization of the Gaussian topoisomers' distribution. It was observed that the twist energy parameter for the given solutes is almost linearly dependent on the bulk dielectric constant. In the reaction buffer, the twist energy parameter was determined to be 1100 +/- 100. By decreasing the dielectric constant to 74 with the addition of sorbitol, the value of the parameter reaches K = 900 +/- 100, whereas the addition of ethylene glycol leads to kappa = 400 +/- 50. Upon addition of glycine, which resulted in a dielectric constant equal to 91, the value of the twist energy parameter increased to K 1750 +/- 100. (c) 2007 Wiley Periodicals.