Adenilson de Souza da Fonseca

Low-level infrared laser effect on plasmid DNA

Low-level laser therapy is used in the treatment of many diseases based on its biostimulative effect. However, the photobiological basis for its mechanism of action and adverse effects are not well understood. The aim of this study, using experimental models, was to evaluate the effects of laser on bacterial plasmids in alkaline agarose gel electrophoresis and Escherichia coli cultures. The electrophoretic profile of bacterial plasmids in alkaline agarose gels were used for studying lesions in DNA exposed to infrared laser. Transformation efficiency and survival of Escherichia coli AB1157 (wild-type), BH20 (fpg/mutM-), BW9091 (xth-), and DH5αF’Iq (recA-) cells harboring pBSK plasmids were used as experimental models to assess the effect of laser on plasmid DNA outside and inside of cells. Data indicate low-level laser: (1) altered the electrophoretic profile of plasmids in alkaline gels at 2,500-Hz pulsed-emission mode but did not alter at continuous wave, 2.5- and 250-Hz pulsed-emission mode; (2) altered the transformation efficiency of plasmids in wild-type and fpg/mutM-E. coli cells; (3) altered the survival fpg/mutM-, xthA- and recA-E. coli cultures harboring pBSK plasmids. Low-level infrared laser with therapeutic fluencies at high frequency in pulsed-emission modes have effects on bacterial plasmids. Infrared laser action can differently affect the survival of plasmids in E. coli cells proficient and deficient in DNA repair mechanisms, therefore, laser therapy protocol should take into account fluencies, frequencies and wavelength of laser, as well as tissue conditions and genetic characteristics of cells before beginning treatment.

Lung morphometry and MMP-12 expression in rats treated with intraperitoneal nicotine.

Nicotine, a toxic tobacco component, plays an important role in the development of cardiovascular and lung diseases in smokers. Our objective was to investigate the effects of the intraperitoneal (i.p.) nicotine treatment in lung morphology. Wistar male rats (3-4 months old) were divided in five groups, a control one, and other groups treated with nicotine (1 mg/kg/day) for 8 days and sacrificed after 24, 48, 96, and 192 h. Morphometry was used to estimate the lung alveolar parenchyme and septal elastic fibers changes, and immunohistochemistry was performed to detect macrophage metalloelastase (MMP-12) and quantify vessels by immunolabelling with alpha-smooth muscle cells. Thickening of the alveolar septa was present in all nicotine groups, and associated with mononuclear cell infiltration, angiogenesis, and irregular areas of collapse. After 96 h, rat lungs showed macrophage, expressing MMP-12, that was also present after 192 h of recovery. Pleural and parenchyma inflammation, fibrosis and macrophage were also seen after 192 h. Intraperitoneal nicotine treated rats exhibited an increase of the volume fraction of alveolar parenchyme, a reduction of volume and surface fraction septal elastic fibers, and an increase of the numerical fraction of microvasculature vessels compared to control ones. MMP-12 was detected in groups of macrophages Wistar rats lung exhibited a progressive morphological damage after 192 hours of recovery, after 8 daily doses of 1 mg/kg body weight on i.p. nicotine.

Low-intensity infrared laser increases plasma proteins and induces oxidative stress in vitro.

Low-intensity laser therapy is based on the excitation of endogenous chromophores in biotissues and free-radical generation could be involved in its biological effects. In this work, the effects of the low-intensity infrared laser on plasma protein content and oxidative stress in blood from Wistar rats were studied. Blood samples from Wistar rats were exposed to low-intensity infrared laser in continuous wave and pulsed-emission modes at different fluencies. Plasma protein content and two oxidative stress markers (thiobarbituric acid-reactive species formation and myeloperoxidase activity) were carried out to assess the effects of laser irradiation on blood samples. Low-intensity infrared laser exposure increases plasma protein content, induces lipid peroxidation, and increases myeloperoxidase activity in a dose- and frequency-dependent way in blood samples. The low-intensity infrared laser increases plasma protein content and oxidative stress in blood samples, suggesting that laser therapy protocols should take into account fluencies, frequencies, and wavelengths of the laser before beginning treatment.

Low intensity infrared laser induces filamentation in Escherichia coli cells

Low intensity continuous wave and pulsed emission modes laser is used in treating many diseases and the resulting biostimulative effect on tissues has been described, yet the photobiological basis is not well understood. The aim of this wok was to evaluate, using bacterial filamentation assay, effects of laser on Escherichia coli cultures in exponential and stationary growth phase. E. coli cultures, proficient and deficient on DNA repair, in exponential and stationary growth phase, were exposed to low intensity infrared laser, aliquots were spread onto microscopic slides, stained by Gram method, visualized by optical microscopy, photographed and percentage of bacterial filamentation were determined. Low intensity infrared laser with therapeutic fluencies and different emission modes can induce bacterial filamentation in cultures of E. coli wild type, fpg/mutM, endonuclease III and exonuclease III mutants in exponential and stationary growth phase. This study showed induction of bacterial, filamentation in E. coli cultures expose to low intensity infrared laser and attention to laser therapy protocols, which should take into account fluencies, wavelengths, tissue conditions, and genetic characteristics of cells before beginning treatment.

Effects of Passiflora edulis flavicarpa on the radiolabeling of blood constituents, morphology of red blood cells and on the biodistribution of sodium pertechnetate in rats

The aim of this study was to evaluate possible effects of Passiflora edulis flavicarpa (P. flavicarpa) extract on the labeling of blood constituents with (99m)Tc, on the morphology of red blood cells, and on the biodistribution of sodium pertechnetate (sodium (99m)Tc). Male Wistar rats were treated with either P. flavicarpa extract or 0.9% NaCl. After that, radiolabeling of blood constituents, morphological analysis of red blood cells and biodistribution of sodium (99m)Tc was evaluated. Radiolabeling of blood constituents and shape of red blood cells were not modified, but a significant (p<0.05) alteration of the biodistribution of sodium (99m)Tc was observed after treatment with P. flavicarpa extract. Although our results were obtained with animals, they could contribute to reduce the risk of misdiagnosis and/or repetition of the examinations in nuclear medicine.

Assessment of Effects of a Cordia salicifolia Extract on the Radiolabeling of Blood Constituents and on the Morphology of Red Blood Cells

Effects of a Cordia salicifolia (porangaba) extract on the labeling of blood cells (BCs) with technetium-99m ((99m)Tc) and on the morphology of red BCs were evaluated. Labeling of cellular and molecular structures with (99m)Tc depends on a reducing agent. Some physical characteristics, as visible absorbance spectrum, electric conductivity, and refractive index of this porangaba extract, were also determined. Blood samples from Wistar rats were incubated with porangaba extract or with 0.9% NaCl (control). Labeling of blood constituents with (99m)Tc was performed. Plasma (P) and BCs, both soluble (SF-P and SF-BC) and insoluble (IF-P and IF-BC) fractions, were separated. The radioactivity in each fraction was counted, and the percentage of radioactivity incorporated (%ATI) was calculated. Blood smears were prepared, fixed, and stained, and the morphology of the red BCs was evaluated. Data showed an absorbance peak at 480 nm and electric conductibility and refractive index concentration-dependent. Porangaba extract decreased significantly (P < .05) the BC, IF-P, and IF-BC %ATI, and no modifications were verified on the shape of red BCs. Analysis of the results reveals that some physical parameters could be useful to aid in characterizing the extract studied. Moreover, it is possible that chemical compounds of this extract could have chelating/redox actions or be capable of binding to plasma and/or cellular proteins.

Comet assay to determine DNA damage induced by food deprivation in rats

The aim of this work was to evaluate, by comet assay, the possible inducing of DNA lesions in peripheral blood mononuclear cells of rats subjected to acute or chronic food deprivation. Wistar male rats were subjected to 72 h of partial (50%), or total acute food deprivation, and then allowed to recover for different time periods (24, 48 and 72 h). In other experiments, comet scores were determined in peripheral blood mononuclear cells of rats subjected to chronic food deprivation (25% and 50%) for 50 days. Blood aliquots were obtained before, during and after food deprivation. Comet assay was carried out, the comet units photographed and scored (class 0 up to 3). Acute and chronic food-deprived rats presented peripheral blood mononuclear cells with DNA lesions (comet classes 1, 2 and 3) and a significant increase (p<0.05) in the number of comet units compared with its basal level. The increase was proportional to acute food deprivation time, but after being taken off, it progressively returned to basal level after 48 h (partial group) or 72 h (total group). Chronic food-deprived rats presented a progressive increase of comet score up to 5 days, and a decrease thereafter to reach a basal level. Possible mechanisms of DNA lesions are discussed.

A Chrysobalanus icaco extract alters the plasmid topology and the effects of stannous chloride on the DNA of plasmids

Folhas de Chrysobalanus icaco (C. icaco) sao usadas na medicina popular (conhecido como Abajeru no Brasil) para controlar a glicemia em pacientes diabeticos. Cloreto estanoso (SnCl2) e um agente redutor potente usado para diferentes propostas e apresenta efeitos citotoxico e genotoxico. O objetivo deste trabalho foi investigar os efeitos de um extrato aquoso de C. icaco na topologia de DNA plasmidial e nos efeitos do cloreto estanoso sobre o DNA plasmidial. Plasmidios pBSK foram incubados com um extrato de C. icaco na presenca ou ausencia do SnCl2 (200 mg/mL), em seguida, o procedimento de eletroforese em gel de agarose foi realizado. Plasmidios incubados somente com SnCl2 foram usados como controle positivo e, como controle negativo, plasmidios incubados com tampao Tris. Os geis foram corados com brometo de etidio e as bandas de DNA foram semiquantificadas por densitometria. Os dados mostraram que o extrato de C. icaco altera o perfil eletroforetico e diminui significativamente (p < 0,05) os efeitos do SnCl2 sobre DNA plasmidial. Os resultados obtidos neste trabalho indicam uma acao protetora dependente da dose e um efeito genotoxico de extrato de C. icaco sobre o DNA plasmidial.

Validation of a new computerized system for recording and analysing drug-induced tremor in rats.

INTRODUCTION Certain drugs can induce tremor in small animals and can be used as Parkinsons disease or essential experimental tremor models. However, the use of arbitrary scales for evaluating tremor in experimental models is limited by observer subjectivity. Progress in electronics and computer science has allowed a more precise quantification of tremor. The objective of the present study was to validate a newly developed low-cost method of spectral registration and analysis of tremor in free-moving rats. METHODS Male Wistar rats, 3-4 months of age, previously placed for 5 min inside a sensor cage, were administered with different doses of eserine (0.25-1.5 mg/kg), oxotremorine (0.25-1.5 mg/kg) or harmaline (7.5-60 mg/kg). Drug-induced tremor was recorded during 10 min using a computerized system composed of force transducers, a signal conditioning circuit, a digitizing interface and a microcomputer. The signal transmitted to the computer was quantified, stored and analyzed for its amplitude and frequency by means of specific programs. RESULTS Tremor was induced with an amplitude that was dose-dependent for all drugs used. Tremor frequency was dose-dependent for oxotremorine and eserine, but not for harmaline. The performance of the system was compared with that of other systems described in behavioral instrumentation literature. DISCUSSION The present data indicate that the new system is capable of detecting the tremor induced by drugs, and that the programs used for spectral analysis allow the quantification of the amplitude and the frequency of the tremor in free-moving rats.

Sucralose sweetener in vivo effects on blood constituents radiolabeling, red blood cell morphology and radiopharmaceutical biodistribution in rats.

Effects of sucralose sweetener on blood constituents labelled with technetium-99m ((99m)Tc) on red blood cell (RBC) morphology, sodium pertechnetate (Na(99m)TcO(4)) and diethylenetriaminepentaacetic acid labeled with (99m)Tc ((99m)Tc-DTPA) biodistribution in rats were evaluated. Radiolabeling on blood constituents from Wistar rats was undertaken for determining the activity percentage (%ATI) on blood constituents. RBC morphology was also evaluated. Na(99m)TcO(4) and (99m)Tc-DTPA biodistribution was used to determine %ATI/g in organs. There was no alteration on RBC blood constituents and morphology %ATI. Sucralose sweetener was capable of altering %ATI/g of the radiopharmaceuticals in different organs. These findings are associated to the sucralose sweetener in specific organs.