Prevention of bacterial translocation using β-(1-3)-D-glucan in small bowel ischemia and reperfusion in rats

To investigate the role of β-(1-3)-D-glucan on 99mTc labelled Escherichia coli translocation and cytokines secretion in rats submitted to small bowel ischemia/reperfusion injury. Methods: Five groups (n=10 each) of Wistar rats were subjected to control(C), sham(S), group IR subjected to 45 min of bowel ischemia/60 min of reperfusion(I/R), and group I/R+glucan subjected to 45 min of bowel ischemia/60 min of reperfusion(I/R) and injected with 2mg/Kg intramuscular. Translocation of labelled bacteria to mesenteric lymph nodes, liver, spleen, lung and serum was determined using radioactivity/count and colony forming units/g(CFU/g). Serum TNFα, IL-1β, IL-6, IL-10 were measured by ELISA. Results: CFU/g and radioactivity/count were higher in I/R than in I/R+glucan rats. In C, S and S+glucan groups, bacteria and radioactivity/count were rarely detected. The I/R+glucan rats had enhancement of IL-10 and suppressed production of serum TNFα, IL-1β and, IL-6, compared to I/R untreated animals. Conclusion: The β-(1-3)-D-glucan modulated the production of pro-inflammatory and anti-inflammatory cytokines during bowel ischemia/reperfusion, and attenuated translocation of labelled bacteria

Effects of simvastatin in abdominal sepsis in rats

Statins are widely recognized as hypolipemic drugs, but some studies have observed anti-inflammatory and immunomodulatory effects, known as pleiotropic. The aims of this work was to study possible anti-inflammatory effects of simvastatin in abdominal sepsis. Serum pro-inflammatory cytokines and leukocytes count were determined in an experimental model of abdominal sepsis, using cecal ligation and puncture (CLP) in rats. Methods: Twenty eigth Wistar rats weighing 285±12g were randomly divided in: CLP/Sinvastatin rats (n=7), treated with 10 mg/Kg of oral simvastatin 18 and 2 hs berofe CLP; CLP/Saline group rats (n=7), treated with oral saline; group Sham/Simvastatin (n=7), treated with simvastatin, and group Sham/Saline (n=7), treated with saline. Serum TNF-α, IL-1β and IL-6 by ELISA and total leukocytes, neutrophils, lymphocytes, and eosinophils were determined 24 hs after CLP. ANOVA and Tukey test were used considering significant p<0.05. Results: It was demonstrated that serum TNF-α, IL-1β and IL-6 were respectively 364,8±42pg/mL; 46,3±18pg/mL and 28,4±13pg/mL in CLP/Sinvastatin rats, significantly lower (p<0.05) than in group CLP/Saline (778,5±86pg/ml; 176,9±46pg/ ml; 133,6±21 pg/ml, respectively). The same results were observed in total leukocytes and neutrophils counts. Conclusion: These results clearly demonstrate that simvastatin is an effective agent that reduces cytokines levels and leukocyte count in sepsis, independently of its well-known lipid-lowering effects. Thus, HMG-CoA reductase inhibitors like simvastatin have important anti-inflammatory effects in abdominal sepsis in rats

Total gastrectomy with substitution of stomach by jejunal pouch with and without duodenal passage. Study in rats

A comparison was done between the F. Paulino jejunal pouch (FP) and a jejunal pouch (JP) as esophagusduodenum interpositional graft, for replacing the stomach after total gastrectomy. It was investigated the effect of the two procedures on esophagus histology, nutritional state and serum gastrin in rats. Methods: Male Wistar rats weighing 282±17g were randomly submitted to sham operation (S), FP and JP after total gastrectomy. After eight weeks the rats were killed with overdose of anesthetic and tissue was taken from the distal esophagus for histology. Serum levels of total proteins, albumin, iron, transferring, folate, cobalamine, calcium, as well as serum gastrin were determined. Survival was considered. Results: Fourty six rats were operated and thirty survived for eight weeks. Five (33.3%) died after FP and 11 (52.3%) after JP (p<0.05). Postoperative esophagitis occurred in 6 JP rats. At 8th week, no difference was observed on body weight when compared FP and JP rats (p>0.05). The JP rats had a significant decrease in serum albumin, glucose, transferrin, iron, folate and calcium, compared to sham (p<0.05). Serum gastrin, iron and calcium were significantly higher in JP rats than in FP rats (p<0.05). In FP rats, transferrin and cobalamine showed significant decrease comparing the preoperative with 8th week levels (p<0.05). Conclusion: F. Paulino pouch in rats had lower mortality than JP, and esophagitis was not detected in it. JP rats had serum gastrin, iron and calcium unaffected, possibly because of preservation of duodenal passage

Influence of Splenectomy on the Biodistribution of Technetium-99m Dimercaptosuccinic Acid (99mTc-DMSA) in Rats

This study aimed to evaluate if the splenectomy alters the biodistribution of 99mTc-DMSA and renal function in Wistar rats. The animals were separated in the groups: splenectomy (n = 6) and control (n = 6). After splenectomy (15 days), the administration of 0.1ml of 99mTc-DMSA IV (0.48 MBq) was carried out. Thirty minutes later, kidney, heart, lung, thyroid, stomach, bladder and femur and samples of blood were isolated. The organs were weighed, counted and the percentage of radioactivity -g (%ATI-g) determined. Serum urea and creatinine, hematocrit, leukocytes and platelets were measured. Statistics by t test (p<0.05) was done. There was a significant reduction in %ATI-g in kidney and blood (p<0.05) of splenectomized animals, a significant increase (p<0.05) of urea (88.8 ± 18.6 mg-dL) and creatinine (0.56 ± 0.08 mg-dL), compared to the controls (51.5±1.6, 0.37±0.02mg-dL, respectively), as well as increase in platelets and leucocytes, and hematocrit reduction. The analysis of the results indicates that in rats, splenectomy seems to alter the renal function and the uptake of 99mTc-DMSA

Biodistribution of the radiophamarceutical sodium pertechnetate (Na99mTcO4) after massive small bowel resection in rats

To evaluate the biodistribution of sodium pertecnetate (Na99mTcO4) in organs and tissues, the morphometry of remnant intestinal mucosa and ponderal evolution in rats subjected to massive resection of the small intestine. Methods: Twenty-one Wistar rats were randomly divided into three groups of 7 animals each. The short bowel (SB) group was subjected to massive resection of the small intestine; the control group (C) rats were not operated on, and soft intestinal handling was performed in sham rats. The animals were weighed weekly. On the 30th postoperative day, 0.l mL of Na99mTcO4, with mean activity of 0.66 MBq was injected intravenously into the orbital plexus. After 30 minutes, the rats were killed with an overdose of anesthetic, and fragments of the liver, spleen, pancreas, stomach, duodenum, small intestine, thyroid, lung, heart, kidney, bladder, muscle, femur and brain were harvested. The biopsies were washed with 0.9% NaCl.,The radioactivity was counted using Gama Counter WizardTM 1470, PerkinElmer. The percentage of radioactivity per gram of tissue (%ATI-g) was calculated. Biopsies of the remaining jejunum were analysed by HE staining to obtain mucosal thickness. Analysis of variance (ANOVA) and the Tukey test for multiple comparisons were used, considering p<0.05 as signifi cant. Results: There were no signifi cant differences in %ATI-g of the Na99mTcO4 in the organs of the groups studied (p>0.05). An increase in the weight of the SB rats was observed after the second postoperative week. The jejunal mucosal thickness of the SB rats was signifi cantly greater than that of C and sham rats (p<0.05). Conclusion: In rats with experimentally-produced short bowel syndrome, an adaptive response by the intestinal mucosa reduced weight loss. The biodistribution of Na99mTcO4 was not affected by massive intestinal resection, suggesting that short bowel syndrome is not the cause of misleading interpretation, if an examination using this radiopharmaceutical is indicated

Prevention of bacterial translocation using β-(1-3)-D-glucan in small bowel ischemia and reperfusion in rats

To investigate the role of β-(1-3)-D-glucan on 99mTc labelled Escherichia coli translocation and cytokines secretion in rats submitted to small bowel ischemia/reperfusion injury. Methods: Five groups (n=10 each) of Wistar rats were subjected to control(C), sham(S), group IR subjected to 45 min of bowel ischemia/60 min of reperfusion(I/R), and group I/R+glucan subjected to 45 min of bowel ischemia/60 min of reperfusion(I/R) and injected with 2mg/Kg intramuscular. Translocation of labelled bacteria to mesenteric lymph nodes, liver, spleen, lung and serum was determined using radioactivity/count and colony forming units/g(CFU/g). Serum TNFα, IL-1β, IL-6, IL-10 were measured by ELISA. Results: CFU/g and radioactivity/count were higher in I/R than in I/R+glucan rats. In C, S and S+glucan groups, bacteria and radioactivity/count were rarely detected. The I/R+glucan rats had enhancement of IL-10 and suppressed production of serum TNFα, IL-1β and, IL-6, compared to I/R untreated animals. Conclusion: The β-(1-3)-D-glucan modulated the production of pro-inflammatory and anti-inflammatory cytokines during bowel ischemia/reperfusion, and attenuated translocation of labelled bacteria

Efeito do consumo de farinha de linhaça (Linum usitatissimum) no crescimento de ratos Wistar e relação com a digestibilidade de globulinas e fatores antinutricionais protéicos

Linseed is an important oilseed consumed raw as nutritional supplement, that although represents a rich source of nutrients, its nutritional value could be impaired due to the presence of antinutritional factors. In this study, protein fractions from raw linseed flour were extracted and isolated being obtained 12% of albumins, 82% of globulins, 5% of glutelins and 1% of prolamins. These proteins were visualized by SDS-PAGE and albumins showed low molecular mass protein bands around 21 kDa and minor bands, similar to that of trypsin inhibitor; Globulins presented protein bands with high molecular masses, which possibly are constituents of multimeric proteins, such as legumins. After determination of the centesimal composition of raw linseed, it was used as exclusive protein source for young rats to evaluate its effect on animal growth. The results showed negative effects on rat growth (weight gain 73% less than the control group) and reduction of intestinal villus (35%), that could be related with in vitro and in vivo globulin digestibility and proteinaceous antinutritional factors (mammalian digestive enzymes inhibitors and lectins) in albumin fraction. Native globulins showed, by SDS-PAGE, low susceptibility in vitro to trypsin and chymotrypsin, however presented high degradation by pancreatin. Thermal treatment of globulins for 5 and 15 minutes at 100ºC improved considerably its digestibility by trypsin and pancreatin. Globulins presented 93.2% in vivo digestibility, similar to the control protein. Albumin fraction had high trypsin inhibition activity (100%) and chymotrypsin inhibition of 28.3%; haemagglutinating activity was not detected. The results of this study indicate the negative action of trypsin inhibitors on animal growth, but can not be discarded its combined action with other antinutritional factors, which could compromise the raw linseed utilization as an alternative food

Avaliação bioquímica e hostológica de fígado de ratas wistar diabéticas e tratadas com tamoxifeno

Tamoxifen (TX), a drug used in the treatment of breast cancer, may cause hepatic changes in some patients. The consequences of its use on the liver tissues of rats with or without diabetes mellitus (DM) have not been fully explored. The purpose of this multidisciplinary study was to evaluate the correlation between plasma hepatic enzyme levels and the presence of iron overload in the hepatic tissue of female Wistar rats with or without streptozotocin-induced DM and using TX. Female rats were studied in control groups: C-0 (non-drug users), C-V (sorbitol vehicle only) and C-TX (using TX). DM (diabetic non-drug users) and DM-TX (diabetics using TX) were the test groups. Sixty days after induced DM, blood samples were collected for glucose, alanine aminotransferase (ALT), aspartate aminotransferase (AST) alkaline phosphatase (ALP) and bilirubin measures. Hepatic fragments were processed and stained with hematoxylin and eosin (H&E), Masson s trichrome, Perls. The hepatic iron content was quantified by atomic absorption spectrometry. AST, ALT and ALP levels were significantly elevated in the DM and DM-TX groups, with unchanged bilirubin levels. Liver iron overload using Perls stain and atomic absorption spectrometry were observed exclusively in groups C-TX and DM-TX. There was positive correlation between AST, ALT and ALP levels and microscopic hepatic siderosis intensity in group DM-TX. In conclusion, TX administration is associated with liver siderosis in diabetic and non-diabetic rats. In addition, TX induced liver iron overload with unaltered hepatic function in 2 non-diabetic rats and may be a useful tool for investigating the biological control of iron metabolism

Efeito de preparações de produtos naturais (cravo da índia e tomate) e de um produto sintético na marcação de constituintes sangüineos com tecnécio-99m e na morfologia de hemácias isoladas de sangue de ratos wistar

There are strong interests in the evaluation of the biological effects of natural and synthetic products. Blood constituents labeled with technetium-99m (99mTc) are used in nuclear medicine. The aim of this work was to study the effects of Clove (Caryophyllus aromaticas L.) and OZE (preparation used in the Health Sciences) on the labeling blood constituents with 99mTc and on the morphologic red blood cells (RBC) and the action of an extract of tomato (TO) on the labeling of blood constituents Blood samples were incubated with clove or OZE or TO, stannous chloride and 99mTc. Plasma (P), blood cells (BC), insoluble fractions (IF) of plasma and blood cells were separated. The radioactivity was counted and percentage of radioactivity (%ATI) to each blood fraction was calculated. The shape and morphometric parameter (perimeter/area ratio) were evaluated in the studies with clove and OZE. Clove extract and OZE altered significantly (p<0.05) the %ATI of blood constituents and the shape of red blood cells. However, clove extract not altered the red blood cells perimeter/area ratio. The tomato extract used at the highest concentrations reduced significantly (p<0.05) the %ATI in IF-P, although this extract did not modify the radiolabeling on BC, neither the radioactivity fixation on IFBC. The results indicate that these chemical compounds would have oxidative/chelating actions

Efeitos de um extrato aquoso de buzhong yi qi wan (fórmula magistral chinesa) na marcação de constituintes sanguíneos com tecnécio-99m, na morfologia e na fragilidade osmótica de hemácias de ratos Wistar

Technetium-99m (99mTc) has been used to obtain several radiobiocomplexes utilized to aid in the diagnosis of diseases. Blood constituents, as red blood cells (RBC) and plasma proteins, have been labeled with 99mTc. Natural and synthetic drugs can alter the labeling of these constituents. The aim of this work was to investigate the possibility of a Buzhong YiQi Wan extract to alter (i) the labeling of blood constituents with 99mTc, (ii) the RBC morphology, and (iii) osmotic fragility of RBC withdrawn from Wistar rats. The data showed that the BYQW extract (i) could affect labeling of blood constituintes with 99mTc, (ii) could affect the membrane integrity decreasing the osmotic resistance and (iii) could not alter the shape of RBC. Probably, these findings would be associated with properties of the substances present in the aqueous extract of BYQW. This study has multiple disciplinary aspects in knowledge areas: Radiobiology, Botanic, Phytotherapy and Haematology

Avaliação de efeitos de um extrato aquoso de cinnamomum zeylanicum L. na marcação de constituintes sanguíneos com tecnécio-99m e na morfologia das hemácias de ratos wistar

Radiobiocomplexes are used to obtain images in nuclear medicine and employed in basic research. Blood constituents labeled with technetium-99m (99mTc) have also been employed as radiobiocomplexes and used also experimental model for evaluation of the biological effects of natural or synthetic drugs. The analysis of the morphology and the morphometrics parameters (perimeter/área ratio) can be used to evaluate the effects of drugs upon the structure of the membrane of red blood cells. Cinnamomum zeylanicum (cinnamon) is a spice used as herbal medicine to treat diseases. The aim of this study was to evaluate the effect of in vitro and in vivo treatment with an aqueous cinnamon extract on the labeling of blood constituents with 99mTc and on the morphology of red blood cells from Wistar rats. In the in vitro treatment, isolated blood sample from animals were incubated with cinnamon extract. In the in vivo treatment, blood samples were also withdrawn from animals treated with cinnamon extract. In both cases, the radiolabeling of blood constituents was done. The morphological analysis of red blood cells was also done. As control, blood or animals treated with NaCl 0.9%. The data obtained on the labeling of blood constituents with 99mTc experiments indicated that the in vitro treatment with cinnamon extract was capable to decrease signiicantly (p<0.05) the percentage of radioactivity in cellular compartments and on the fixation of cellular and plasma proteins. These effects were not observed on the in vivo treatment. The results obtained for the morphology of red blood cells suggest that the in vitro and in vivo treatments did not alter the morphology and the perimeter/area ratio. The in vitro treatment with aqueous cinnamon extract could affect the membrane structures related with the oxidation status of the stannous ion pertechnetate ion, altering the labeling of blood constituentes with 99mTc. This study was a multidisciplinary experimental research. It was developed with the contribution of the different Departments and Services of the Hospital Universitário Pedro Ernesto of the Universidade do Estado do Rio de Janeiro

Efeitos de um extrato aquoso de lantana camara (Cambará de espinho) na marcação de constituintes sanguíneos com técnecio-99m e na morfologia de hemácias de ratos-wistar

Medicinal plants have been studied and used in the world. Lantana camara has medicinal properties and it has been used in folk medicine. The aim was to verify the effect of a lantana extract on the labeling of blood constituents with 99mTc, and to evaluate the effect of an aqueous extract of Lantana camara on the morphology of RBC withdrawn from Wistar rats. The results showed that lantana extract has decreased the fixation of radioactivity on the IF-P. This effect was not observed in the BC compartment and in IF-BC. The BC-%ATI was decreased in all concentrations tested when the BC was washed. The osmotic fragility assay and morphological analysis were carried out. In presence of the extract, the data obtained indicated that (i) an increase of the hemolysis and (ii) modifications on the morphology of RBC. These effects of the Lantana camara could be associated with some pharmacological properties of the chemical compounds of this studied extract

Estudo de efeitos de um extrato de Arctium lappa (Bardana) na marcação dos constituintes sangüíneos com Tecnécio-99m, na biodisponibilidade do radiofármaco pertecnatato de sódio e na morfologia de hemácias de ratos Wistar

Radionuclides have been used in Nuclear Medicine for diagnostic and treatment. In basic research, cellular and molecular structures are labeled with technetium-99m (99mTc) and used as radiobiocomplexes. Some natural or synthetic drugs are capable to alter the labeling of blood constituents with 99mTc, as well as in the biodistribution of radiobiocomplexes. Arctium lappa (Bardana) has been used to treat inflammatory processes. The aim of this work was to evaluate the effects of an extract of Bardana on the labeling of blood constituents with 99mTc, on the morphology of red blood cells, on the perimeter/area ratio of red blood cells and on the biodistribution of radiophamaceutical sodium pertechnetate in Wistar rats. Extract of Bardana was capable to alter the labeling of cellular compartment with 99mTc. Plasma and cellular proteins did not present alteration on the percentage of radioactivity (%ATI). Extract of Bardana was also capable to alter the morphology and the perimeter/area ratio of red blood cells. On the biodistribution of sodium pertechnetate in animals treated with the extract of Bardana, it was observed a small and significant uptake in liver, tooth and tongue, and a high and a significant uptake in stomach, lung and testis (p<0.05). In conclusion, these findings could be justified due to the effects of some chemical compounds in the Bardana extract. This study was a multidisciplinary experimental research. It was developed with the contribution of the different Departments and Services of the Hospital Universitário Pedro Ernesto of the Universidade Estadual do Rio de Janeiro

Estudo de efeitos de um extrato aquoso de farinha de casca de maracujá (Passiflora edulis f. flavicarpa) na marcação de constituintes sangüíneos com Tecnécio 99m, na morfometria das hemácias e na biodisponibilidade do radiofármaco Pertecnetato de Sódio em ratos Wistar

Blood constituents labelled with technetium-99m (99mTc) has been used with radiobiocomplexes in several procedures in nuclear medicine. Some natural and sintetic drugs are capable to interfere on the labeling of blood constituents with 99mTc, on the morphology of red blood cells (RBC) and on the biodistribution of radiobiocomplexes. The aim of this study was evaluate the effect of an extract of Passiflora edulis f. flavicarpa on the labeling of blood constituints with 99mTc, on the morphology of RBC and on the biodistribution of the radiopharmaceutical sodium perthecnetate in Wistar rats. On the in vitro studies the Passiflora edulis f. flavicarpa decreased significantly (p<0.05) the %ATI on plasma proteins and on the in vitro morhology of RBC, the passion fruit peel flour altered the shape and the perimeter/área ratio. On the in vivo estudies the extract did not altered the %ATI in blood constituents, and did not altered the shape of RBC. Although, on the biodistribution of the radiobiocomplex sodium perthecnetate (Na99mTcO4) this extract decreased significantly (p<0.05) the uptake in duodenum, spleen, pâncreas and blood, and increased the uptake in stomach. It can be suggested that the effects presented by this extract could be a result of some substances contained in this extract that could alter the binding of 99mTc to plasma proteins, the morphology of RBC and the biodistribution of the radiobiocomplex sodium perthecnetate. This study was multidisciplinary experimental research. It was developed with the contribution of different Departments and Services of the Hospital Universitário Pedro Ernesto of the Universidade Estadual do Rio de Janeiro

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.