Efeito de um extrato de Artemísia Vulgaris L. marcação in vitro de constituintes sanguíneos com tecnécio-99m em ratos wistar

Artemisia vulgaris L..is used in folk medicine and in Traditional Chinese Medicine (TCM). This medicinal plant has been utilized as anticonvulsive, analgesic, antispasmodic effect, rheumatic pains, menstrual dyspepsia, asthenia, epilepsy, hepatitis, fevers, anemia and to expel parasites. In nuclear medicine, blood constituents are labeled with technetium-99m (99mTc) and used as radiopharmaceuticals (radiobiocomplexes). Authors have been described that synthetic and/or natural drugs could modify the labeling of blood constituents with 99mTc. The aim of this work was to evaluate the effects of an aqueous extract of Artemisia vulgaris L. on the labeling of blood constituents with 99mTc. Blood samples withdrawn of Wistar rats were incubated with Artemisia vulgaris L, stannous chloride and 99mTc, as pertechnetate ion. Aliquots of plasma (P) and blood cells (BC) were isolated. Aliquots of P and BC were also precipitated with trichloroacetic acid and soluble (SF) and insoluble (IF) fractions were separated. The radioactivity in each fraction was counted and the percentages of radioactivity (%ATI) were calculated. Artemisia vulgaris L. extract decreased significantly (p<0.05) the %ATI on BC and on IF-BC. The analysis of the results indicates that the extract could have substances that could interfere on the transport of stannous through the erythrocyte membrane altering the labeling of blood cells with 99mTc. Working in this study was a multidisciplinary group, with Phisical therapists, Biomedicals, Physicals, Pharmacists, Biologists, Statistics and Physicians.

Avaliação de efeitos DW extratos de sambucus australis E sambucus nigra na marcação de constituintes sanguineos com tecnécio-99M, na morfologia de Hemácias de ratos wistar, na topologia plasmidial e na ação do cloreto estanoso no DNA Plasmidial

Radiobiocomplexes are used in nuclear medicine to obtain images and to treat diseases. Blood constituents have been used as radiobiocomplexes. Natural or synthetic products can influence on the labeling of blood constituents with technetium-99m (99mTc), the morphology of red blood cells and on the stannous chloride (SnCl2) action on plasmid DNA. Sambucus australis and Sambucus nigra are used in popular culture for treating diseases. The aim of this work was to evaluate the effects of the extracts of Sambucus australis and Sambucus nigra on the labeling of blood constituents with 99mTc, on morphology of red blood cells of Wistar rats, on the topology of plasmids DNA and the action against the SnCl2 effects on the DNA of plasmids pBSK. On the labeling of blood constituents with 99mTc it was verified that both extracts were capable to decrease significantly the radioactivity in the cellular compartment and in the insoluble fraction of plasma. Sambucus australis also decreased the labeling of insoluble fraction of blood cells with 99mTc. Both extracts did not alter the morphology of red blood cells. Moreover, it was verified that Sambucus nigra did not alter the electrophoretic profile of plasmid DNA, but decreased the effect of SnCl2 on plasmid DNA. These last results sugest a genotoxic effect and a protective action of Sambucus nigra extract against the SnCl2 action on plasmid DNA. This work was developed with the contribution of several Departments of biomedical area of the Hospital Universitário Pedro Ernesto, of the UERJ, characterizing a multidisciplinary experimental research

Efeitos de um extrato aquoso de porangaba (Cordia salicifolia) na marcação de constituintes sanguíneos com tecnécio-99m e na morfologia de hemácias

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

Efeito de um extrato de Ganoderma lucidum (Reishi) na marcação de constituintes sanguíneos com tecnécio-99M e na sobrevivência de Escherichia coli

Clinical evaluations have been made possible with radiobiocomplexes marked with tecnecium-99m (99mTc). Natural or synthetic drugs are able to interfere in the marking of blood structures with 99m Tc. Also, the toxicity of several natural products has been described. The aim of this study was evaluating the effect of an extract of Ganoderma lucidum (Reishi) in the marking of blood constituents with 98m Tc and in the survival of Escherichia coli. Blood samples from Wistar rats were treated with reishi extract. Radiomarking procedure was performed. Samples of plasma (P), blood cells (CS), and insoluble (FI) and soluble (FS) fractions of P and CS were separated and the radioactivity was counted to determine radioactivity percentages (%ATI). Escherichia coli AB1157 cultures were treated with stannous chloride in the presence and absence of the reishi extract. Blood samples and bacterial cultures treated with NaCl 0.9% were used as controls. Data indicated that the reishi extract has significantly altered (p<0,05) the %ATI of P, CS, FI-P, FS-P, FI-CS e FS-CS, as well as it has increased survival of bacterial cultures treated with stannous chloride. Our results suggest that the Reishi extract would be able to present a redox/ chelant action by altering blood constituent marking with 99mTc and by protecting bacterial cultures against stannous chloride-induced oxydating lesions. The study had a multidisciplinary character, with the participation of the following areas of knowledge: Biophysics, Radiobiology, Botanics, Phytotherapy, and Hematology

Avaliação de efeitos biológicos de antiinflamatórios derivados do ácido propiônico através de modelos experimentais em nível molecular e celular

Derivatives of propionic acid NSAIDs are irreversible inhibitors of cyclooxygenase enzyme widely used. The aim of this study was to evaluate, through different experimental models, biological effects of derivatives of propionic acid (fenoprofen, naproxen, ibuprofen and ketoprofen) in cellular and molecular level. The labeling of blood constituents with technetium-99m (99mTc) and morphological analysis of erythrocytes of blood of rats, as well as growth, survival of cultures of Escherichia coli (E. coli) and the assessment of bacterial plasmid electrophoretic profiles were models used for experimental evaluation of possible biological effects of antiinflammatory drugs. The results show that, in general, anti-inflammatory drugs evaluated were not able to alter the labeling of blood constituents with 99mTc, the morphology of red blood cells from blood of rats, as well as the growth of cultures of E. coli and the electrophoretic profile of plasmid DNA. However, naproxen appears to cytotoxic effect on bacterial cultures, plasmids and genotoxic effects in reducing the action of stannous chloride in cultures of E. coli. The use of experimental fast performance and low cost was important for assessment of biological effects, contributing to a better understanding of the properties of propionic acid derivatives studied. anti-inflammatory, blood constituents, technetium-99m, stannous chloride, Escherichia coli; DNA

Análise in vitro da atividade antifúngica e de toxicidade da anfotericina B pré-aquecida

The aim of this work was to evaluate how an aqueous micellar system containing Amphotericin B (AmB) and sodium deoxycholate (DOC) can be rebuilt after heating treatment. Also a review of the literature about the new physicochemical and biological properties of this new system was carried out. Afterwards, heated (AmB-DOC-H) and unheated (AmB-DOC) micelles were subsequently diluted at four different concentrations (50mg.L-1, 5mg.L-1, 0.5mg.L-1 and 0.05mg.L-1) to perform the physicochemical study and, then, the pharmacotoxicity assay, in which two cell models were used for the in vitro experiments, Red Blood Cells (RBC) from human donors and Candida parapisilosis (Cp). While potassium (K+) and hemoglobin leakage from RBC were the used parameters to evaluate the acute and chronic toxicity, respectively, the efficacy of AmB-DOC and AmB-DOC-H were assessed by K+ leakage and cell survival rate from Cp. The spectral study revealed a slight change on the aggregate peak from 327nm to 323nm for AmB-DOC-H compared to AmB-DOC. Concerning the toxicity, although AmB-DOC and AmB-DOC-H presented different behavior for hemoglobin leakage, AmB-DOC produced higher leakage than AmB-DOC-H at high concentrations (from 5mg.L-1) with values tending to zero. However, concerning K+ leakage, both AmB-DOC and AmB-DOC-H, showed similar profile for both cell models, RBC and Cp (p<0,05). AmB-DOC-H and AmB-DOC also revealed similar profile of activity against Cp with equivalent survival rate. In short, the AmB-DOC-H showed much less toxicity than AmB-DOC, but remained as active as the late one against fungal cell. Therefore, the results highlight the importance of this new procedure as a simple, inexpensive and safe alternative to produce a new kind of micelle system for treatment of systemic fungal infections

Análise da presença de agentes mutagênicos nas águas do Rio Pitimbú/RN

The Pitimbu River is located at the oriental portion of the State of Rio Grande do Norte, including three importants cities named Macaíba, Parnamirim e Natal. Although its high importance as a water source, which supplys great part of the South Zone of the Natal city, this river receives a large quantity of domestic and industrial waste water without treatment. The Pitimbu River headhas its river-head located in the city of Macaiba, goes through Parnamirim, then it flowing into at the Jiqui Lake in Natal. The aim of this study was to evaluate, qualitatively and quantitatively, the environmental quality of the Pitimbu River by genotoxicity bio-assays, which are important tools for genetics toxicological evaluation. In this work, five samples sites, distributed along the river, were used to collect water samples. Another point site, located near Jiqui lake, was used to collect drinkable water, which was treated by CAERN, the water treatment entreprise of Rio Grande do Norte. The following assays where used to evaluate the quality of these samples: Allium cepa assay; Comet assay; Micronuclei (MN) assay; and Ames test. For the Allium cepa assay, sixteen specimes where used for each water sample from the sample sites. In this assay both microscopic, like cytogenetic damage, and macroscopic aspects, as morphological variation were evaluated. Red blood cells from periferical blood of the Crenicichla menezesi native specie were used not only for the MN assay, but also for the Comet assay. These fishes were collected at different points on the Pitimbu River and the negative control was developed using fishes of the same species that were bring to the laboratory and maintained for 100 days in the optimal experimental conditions. For the Ames test, TA100, YG1042, TA98 and YG1041 strains were used in the directed method without metabolic activation. The results found by the Allium cepa assay showed that two water sample sites induced increase of mitotic index (IM). Additionally, compared to the control, all the water samples increased the chromossomal aberrations frequency and/or micronucleus. Among the sample sites, two also showed an abnormal growth rate in its root and two samples induced morphological alteration. With the MN test in red blood cells, a high frequence of MN was observed in tree sample. By comparing all the results obtained on the water sample points and with the negative control, a significant variation on the MN frequency was observed. Positive results were also observed for the same sample to water test by the Comet assay. These results allow concluding that the proposed specie Crenicichla menezesi has a good profile as a bio-indicator for the evaluation of environmental water quality and the MN and comet test can be usuful for in situ evaluation. By the Ames test, it was possible to detect the mutagenic activity on the waters from the Pitimbu River in different levels of mutagenicity. This result suggests that this river has several substances that induced changes directly to the DNA. The mechanisms involved to this phenomenon could be by both processes, by changing of the reading frame and by nucleotide substitution. These data set indicate the presence of mutagenic agents, which can represent in risk to biot and human beens

Consequências da colectomia associada à hepatectomia no metabolismo hepático e na forma e função de hemácias em ratos

This study investigated the influence of partial colectomy associated with hepatectomy on the biodistribution of the 99mTc-phytate, on metabolic parameters, as well as labeling and morphology of red blood cells. Wistar rats were distributed into three groups (each with 6), nominated as colectomy, colectomy+hepatectomy and sham. In the 30th postoperative day all rats were injected with 99mTc-phytate 0.1mL i.v. (radioactivity 0.66 MBq). After 15 minutes, liver sample was harvested and weighed. Percentage radioactivity per gram of tissue (%ATI/g) was determined using an automatic gamma-counter. Serum AST, ALT, alkaline phosphatase and red blood cells labeling were determined. The liver %ATI/g and red blood cells labeling were lower in colectomy and colectomy+hepatectomy rats than in sham rats (p <0.05), and no difference was detected comparing the colectomy and colectomy+hepatectomy groups. Red blood cells morphology did not differ among groups. Serum levels of AST, ALT and alkaline fosfatase were significantly higher in colectomy+hepatectomy than in colectomy rats (p<0.001). Hepatectomy associated with colectomy lowered the uptake of radiopharmaceutical in liver and in red blood cells in rats, coinciding with changes in liver enzymatic activity

Influência da laparoscopia e da laparotomia na gasometria, leucometria diferencial e citocinas em modelo de sepse abdominal em ratos

Laparoscopic surgery is associated with reduced surgical trauma, and less acute phase response, as compared with open surgery. Cytokines are important regulators of the biological response to surgical and anesthetic stress. The aim of this study was to determine if CO2 pneumoperitoneum would change cytokine expression, gas parameters and leukocyte count in septic rats. Methods: Wistar rats were randomly assigned to five groups: control (anesthesia only), laparotomy, CO2 pneumoperitoneum, cecum ligation and puncture by laparotomy, and laparoscopic cecum ligation and puncture. After 30 min of the procedures, arterial blood samples were obtained to determine leukocytes subpopulations by hemocytometer. TNFα, IL-1β, IL-6 were determined in intraperitoneal fluid (by ELISA). Gas parameters were measured on arterial blood, intraperitoneal and subperitoneal exsudates. Results: Peritoneal TNFα, IL-1β and IL-6 concentrations were lower in pneumoperitoneum rats than in all other groups (p<0.05). TNFα, IL-1β and IL-6 expression was lower in the laparoscopic than in laparotomic sepsis (p<0.05). Rats from laparoscopic cecum ligation and puncture group developed significant hypercarbic acidosis in blood and subperitoneal fluid when compared to open procedure group. Total white blood cells and lymphocytes were significantly lower in laparoscopic cecum ligation and puncture rats than in the laparotomic (p<0.01). Nevertheless, the laparotomic cecum ligation rats had a significant increase in blood neutrophils and eosinophils when compared with controls (p<0.05). Conclusions: This study demonstrates that the CO2 pneumoperitoneum reduced the inflammatory and immune response in an animal model of peritonitis with respect to intraperitoneal cytokines, white blood cell count and clinical correlates of sepsis. The pneumoperitoneum produced hypercarbic acidosis in septic animals

Avaliação da atividade anti-Helicobacter pylori e citotóxica in vitro de extratos orgânicos obtidos das folhas de Encholirium spectabile e Syzygium cumini

Helicobacter pylori is a spiral, Gram negative, mobile, and microaerophilic bacteria recognized as a major cause of gastritis, ulcer, gastric cancer, and gastric low grade, B cell, mucosa – associated lymphoid tissue (MALT) lymphoma, constituting an important microorganism in medical microbiology. Its importance comes from the difficulty of treatment because the requirement of multiple drugs use, besides the increasing emergence of resistant and multiresistant strains to antibiotics used in th e clinic. In order to expand safe and effective therapeutic options , chemical studies on medicinal plants by obtaining extracts, fractions, isolated compounds or essential oils with some biological activity has been intensified . Given the above, the objective was to evaluate the inhi bitory activity of organic extracts derived from Syzygium cumini and Encholirium spectabile, with antiulcer history, and the essential oil, obtained from S. cumini, against H. pylori (ATCC 43504) by the disk diffusion method, for qualitative evaluation, an d determination of minimum inhibitory concentration (MIC) using the broth microdilution method, for quantitative analysis. Also was evaluated the extracts in vitro toxicity by a hemolytic assay using sheep red blood cells, and VERO and HeLa cells using the MTT assay to analyze cell viability. The extracts of both plant used in antimicrobial assays did not inhibit bacterial growth, however the essential oil of S. cumini (SCFO) proved effective, showing MIC value of 205 μg/mL (0.024 % dilution of the original oil). In the hemolytic assay, the same oil shows moderate toxicity, by promote 25% hemolysis at 1000 μg/mL. Regarding the cytotoxicity in cell culture, the SCFO, at 260 μg/mL, affected the cell viability around 80% of HeLa and 50% of VERO cells. So the oi l obtained from S. cumini leaves has antimicrobial activity against H. pylori and cytotoxicity potential, suggesting a source of new molecule drug candidates, since new stages of toxicity in vitro and in vivo, as well, chemical characterization be evaluate d. Moreover, the development of a prospective drug delivery system can result in a prototype to be used in preclinical tests.

Sistema de contagem com morfologia matemática fuzzy

Mathematical Morphology presents a systematic approach to extract geometric features of binary images, using morphological operators that transform the original image into another by means of a third image called structuring element and came out in 1960 by researchers Jean Serra and George Matheron. Fuzzy mathematical morphology extends the operators towards grayscale and color images and was initially proposed by Goetherian using fuzzy logic. Using this approach it is possible to make a study of fuzzy connectives, which allows some scope for analysis for the construction of morphological operators and their applicability in image processing. In this paper, we propose the development of morphological operators fuzzy using the R-implications for aid and improve image processing, and then to build a system with these operators to count the spores mycorrhizal fungi and red blood cells. It was used as the hypothetical-deductive methodologies for the part formal and incremental-iterative for the experimental part. These operators were applied in digital and microscopic images. The conjunctions and implications of fuzzy morphology mathematical reasoning will be used in order to choose the best adjunction to be applied depending on the problem being approached, i.e., we will use automorphisms on the implications and observe their influence on segmenting images and then on their processing. In order to validate the developed system, it was applied to counting problems in microscopic images, extending to pathological images. It was noted that for the computation of spores the best operator was the erosion of Gödel. It developed three groups of morphological operators fuzzy, Lukasiewicz, And Godel Goguen that can have a variety applications

Moléculas coestimulatórias na leishmaniose visceral

Visceral leishmaniasis (VL) is endemic in many countries, including Brazil. The protozoan Leishmania infantum, is the etiological agent of VL, and is transmitted by the bite of female sandflies during the blood meal. The majority of subjects when exposed to the parasite do not develop the disease, because of development of Th1 cellular responses. Those who have develop signs of VL such as fever, weight loss, hepatosplenomegaly, have impairment of the cellular immune response, specific to the Leishmania antigens. We evaluated whether the specififc anergy during symptomatic VL, may be associated with changes in T cells costimulatory molecules or their ligands in CD14+ monocytes. There is an increase in CTLA-4 porcentage on CD4+ T lymphocytes (p=0.001) and ICOS on CD4+ and CD8+ T cells (p=0.002 to CD4+ and p=0.003 to CD8+), after stimulation by soluble Leishmania antigen (SLA) during active visceral leishmaniasis, and that there is a higher percentage of these molecules ex vivo, when comparing symptomatic to recovered individuals (p=0.04 to CTLA-4 in CD4+, and p=0.001 to ICOS in CD4+ and p=0.026 to CD8+). Moreover, we found a high gene expression of CTLA-4, OX-40 and ICOS during active VL. CD40, CD80, CD86, HLA-DR and ICOSL molecules do not suffer changes during disease. There is IFN-γ production by the peripheral blood cells, after SLA stimulation, by peripheral blood cells in symptomatic subjects; however, there is a decrease of the ratio IFN-γ/IL-10, which is reversed after clinical recovery. The impairment of some costimulatory molecules pathways during symptomatic VL could inhibit the ability of phagocytes to kill Leishmania and could facilitate their survival and the proliferation inside macrophages.

The ileum as a determinant organ of the functional liver cell mass in rats

PURPOSE: To evaluate if the ileum resection changes the functioning liver cell mass, the hepatic metabolism and the biodistribution of radiopharmaceutical in rats. METHODS: Twelve Wistar rats weighing 285g±34g were randomly divided into the ileum resection group (n = 6) and sham group rats (n = 6). After 30 days, they were anesthetized and 0.1mL of 99m-Tc-phytate(0.66MBq) was injected via femoral vein. After 30 minutes, blood samples were collected for red blood cells radioactive labeling and serum ALT, AST and gammaGT. Liver samples were used for 99m-Tc-phytatepercentage of radioactivity/gram of tissue and histopathology. Student’s t test was used with significance 0.05. RESULTS: There was a higher uptake of 99m-Tc-phytate in the liver of sham rats, compared to the ileum resection group (p<0.05). GammaGT, ALT and AST were increased in ileum resection rats compared to sham (p<0.05). The he patocytes count was significantly lower in ileum resection group than in sham (p<0.05). Liver: body mass ratio was lower in experimental animals than in sham group (p<0.05). CONCLUSION: These data support that the ileum has important role in liver function and liver mass regulation, and they have potential clinical implications regarding the pathogenesis of liver injury following lower bowel resection.

Metabolic and hematologic consequences of colectomy associated to hepatectomy in rats

To investigate the influence of partial colectomy associated with hepatectomy on the biodistribution of the 99mTc-phytate, on metabolic parameters, as well as labeling and morphology of red blood cells. METHODS: Wistar rats were distributed into three groups (each with six), nominated as colectomy, colectomy+hepatectomy and sham. In the 30th postoperative day all rats were injected with 99mTc-phytate 0.1mL i.v. (radioactivity 0.66 MBq). After 15 minutes, liver sample was harvested and weighed. Percentage radioactivity per gram of tissue (%ATI/g) was determined using an automatic gammacounter. Serum AST, ALT, alkaline phosphatase and red blood cells labeling were determined. RESULTS: The liver %ATI/g and red blood cells labeling were lower in colectomy and colectomy+hepatectomy rats than in sham rats (p <0.05), and no difference was detected comparing the colectomy and colectomy+hepatectomy groups. Red blood cells morphology did not differ among groups. Serum levels of AST, ALT and alkaline fosfatase were significantly higher in colectomy+hepatectomy than in colectomy rats (p<0.001). CONCLUSION: Hepatectomy associated with colectomy lowered the uptake of radiopharmaceutical in liver and in red blood cells in rats, coinciding with changes in liver enzymatic activity.

Influence of laparoscopy and laparotomy on gasometry, leukocytes and cytokines in a rat abdominal sepsis model

Laparoscopic surgery is associated with reduced surgical trauma, and less acute phase response, as compared with open surgery. Cytokines are important regulators of the biological response to surgical and anesthetic stress. The aim of this study was to determine if CO2 pneumoperitoneum would change cytokine expression, gas parameters and leukocyte count in septic rats. Methods: Wistar rats were randomly assigned to five groups: control (anesthesia only), laparotomy, CO2 pneumoperitoneum, cecum ligation and puncture by laparotomy, and laparoscopic cecum ligation and puncture. After 30 min of the procedures, arterial blood samples were obtained to determine leukocytes subpopulations by hemocytometer. TNFα, IL-1β, IL-6 were determined in intraperitoneal fluid (by ELISA). Gas parameters were measured on arterial blood, intraperitoneal and subperitoneal exsudates. Results: Peritoneal TNFα, IL-1β and IL-6 concentrations were lower in pneumoperitoneum rats than in all other groups (p<0.05). TNFα, IL-1β and IL-6 expression was lower in the laparoscopic than in laparotomic sepsis (p<0.05). Rats from laparoscopic cecum ligation and puncture group developed significant hypercarbic acidosis in blood and subperitoneal fluid when compared to open procedure group. Total white blood cells and lymphocytes were significantly lower in laparoscopic cecum ligation and puncture rats than in the laparotomic (p<0.01). Nevertheless, the laparotomic cecum ligation rats had a significant increase in blood neutrophils and eosinophils when compared with controls (p<0.05). Conclusions: This study demonstrates that the CO2 pneumoperitoneum reduced the inflammatory response in an animal model of peritonitis with respect to intraperitoneal cytokines, white blood cell count and clinical correlates of sepsis. The pneumoperitoneum produced hypercarbic acidosis in septic animals