Efecto antiiflamatorio de la fracción flavonoide de Lepechinia meyenii (Walp.) Epling (salvia) sobre leucocitos de pacientes con artritis reumatoide / Anti-Inflammatory Effect of The Flavonoid Fraction of Lepechinia meyenii (Walp.) Epling (SAGE) on Leukocytes of Patients With Rheumatoid Arthritis

RESUMEN Objetivos . Evaluar el efecto antiinflamatorio de la fracción flavonoide de Lepechinia meyenii (Walp.) Epling sobre leucocitos de pacientes con artritis reumatoide (AR). Materiales y métodos. Se recolectaron plantas de la especie Lepechinia meyenii (Walp.) Epling extrayendo diferentes fracciones flavonoides por cromatografía de columna y de capa fina. Se evaluó la producción de anión superóxido mediante la técnica de ensayo reducción nitroblue tetrazolium, en neutrófilos obtenidos de sangre de pacientes con AR, separados en tres grupos: control negativo, que consistió de neutrófilos (5x105 células), control positivo, formado por neutrófilos activados con PMA (phorbol myristate acetate) (150 ng/mL) y los tratamientos, formados por neutrófilos activados y tratados con diferentes concentraciones de la fracción flavonoide LM8 (60, 120 y 180 ug/mL). La expresión de genes proinflamatorios se estudió por RTqPCR, en leucocitos mononucleares obtenidos de pacientes con AR separados en tres grupos: control negativo, que consistió de leucocitos mononucleares (5x105 células), control positivo formado por leucocitos mononucleares activados con fitohemaglutinina (PHA) (150 ug/mL) y el tratamiento formado por leucocitos mononucleares activados y tratados con la fracción flavonoide LM8 (120 ug/mL). Resultados . Se purificaron varias fracciones flavonoides, resultando la fracción LM8 con el mejor efecto inmunomodulador. Dicha fracción disminuyó la producción de anión superóxido en una manera dependiente de la concentración. Por otro lado, disminuyó la expresión de TNFα, IL8 e IL17 en leucocitos mononucleares. Conclusiones. Estos resultados son alentadores respecto al efecto inmunomodulador de esta planta medicinal peruana y justifican continuar su estudio para una posible aplicación clínica.

ABSTRACT Objectives. to assess the anti-inflammatory effect of the flavonoid fraction of Lepechinia meyenii (Walp.) Epling on leukocytes of patients with rheumatoid arthritis (RA). Materials and Methods. Plants of the species Lepechinia meyenii (Walp.) Epling were collected and then different flavonoid fractions were extracted by column and thin layer chromatography. The superoxide anion production was evaluated by means of the reduction of nitroblue tetrazolium assay technique in neutrophils obtained from the blood of patients with RA, divided into three groups: negative control, which consisted of neutrophils (5x105 cells); positive control, made up of PMA (phorbol myristate acetate)-activated neutrophils (150 ng/mL), and the treatments, comprised of neutrophils activated and treated with different concentrations of the flavonoid fraction LM8 (60, 120, and 180 ug/mL). The expression of pro- inflammatory genes was studied by RTqPCR in mononuclear leukocytes obtained from patients with RA, divided into three groups: negative control, which consisted of mononuclear leukocytes (5x105 cells); positive control, made up of phytohaemagglutinin (PHA) (150 ug/ml)-activated mononuclear leukocytes, and the treatment, comprised of mononuclear leukocytes activated and treated with the flavonoid fraction LM8 (120 ug/mL). Results. Several flavonoid fractions were purified, with fraction LM8 showing the best immunomodulating effect. Said fraction diminished the superoxide anion production dependent on concentration. On the other hand, it diminished the expression of TNFα, IL8, and IL17 in mononuclear leukocytes. Conclusions. These results are encouraging in terms of the immunomodulating effect of this Peruvian medicinal plant and justify the continuation of their study for a potential clinical application.

Effects of ethyl pyruvate on leukocyte-endothelial interactions in the mesenteric microcirculation during early sepsis treatment

OBJECTIVES: Experimental studies on sepsis have demonstrated that ethyl pyruvate is endowed with antioxidant and anti-inflammatory properties. This study aimed to investigate the effects of ethyl pyruvate on leukocyte-endothelial interactions in the mesenteric microcirculation in a live Escherichia coli-induced sepsis model in rats. METHODS: Male Wistar rats were administered an intravenous suspension of E. coli bacteria or were subjected to a sham procedure. Three hours after bacterial infusion, the rats were randomized into the following groups: a control group without treatment, a group treated with lactated Ringer’s solution (4 mL/kg, i.v.), and a group treated with lactated Ringer’s solution (4 mL/kg, i.v.) plus ethyl pyruvate (50 mg/kg). At 24 h after bacterial infusion, leukocyte-endothelial interactions were investigated using intravital microscopy, and the expression of P-selectin and intercellular adhesion molecule-1 was evaluated via immunohistochemistry. White blood cell and platelet counts were also determined at baseline and 3 h and 24 h after E. coli inoculation. RESULTS: The non-treated and lactated Ringer’s solution-treated groups exhibited increases in the numbers of rolling leukocytes (∼2.5-fold increase), adherent cells (∼3.0-fold), and migrated cells (∼3.5-fold) compared with the sham group. In contrast, treatment with Ringer’s ethyl pyruvate solution reduced the numbers of rolling, adherent and migrated leukocytes to the levels observed in the sham group. Additionally, the expression of P-selectin and intercellular adhesion molecule-1 was significantly increased on mesenteric microvessels in the non-treated group compared with the sham group (p<0.001). The expression of both adhesion molecules was reduced in the other groups, with ethyl pyruvate being more effective than lactated Ringer’s solution. Infusion of bacteria caused significant leukopenia (3 h), followed ...

Antifungal activity against Cryptococcus neoformans strains and genotoxicity assessment in human leukocyte cells of Euphorbia tirucalli L.

In the last times, focus on plant research has increased all over the world. Euphorbia tirucalli L., a plant known popularly as Aveloz, and originally used in Africa, has been drawing attention for its use in the United States and Latin America, both for use as an ornamental plant and as a medicinal plant. E. tirucalli L. is a member of the family Euphorbiaceae and contains many diterpenoids and triterpenoids, in particular phorbol esters, apparently the main constituent of this plant, which are assumed to be responsible for their activities in vivo and in vitro. The in vitro antifungal activities of Euphorbia tirucalli (L.) against opportunistic yeasts were studied using microbroth dilution assay. The results showed that aqueous extract and latex preparation were effective against ten clinical strains of Cryptococcus neoformans in vitro (Latex and extract MIC range of 3.2 - > 411 µg/mL). Aiming the safe use in humans, the genotoxic effects of E. tirucalli were evaluated in human leukocytes cells. Our data show that both aqueous extract and latex preparation have no genotoxic effect in human leukocytes cells in vitro. Although the results cannot be extrapolated by itself for use in vivo, they suggest a good perspective for a therapeutic application in future. In conclusion, our results show that the aqueous extract and latex preparation from E. tirucalli L. are antifungal agents effectives against several strains of C. neoformans and do not provoke DNA damage in human leukocyte cells, considering the concentrations tested.

Alprostadil attenuates inflammatory aspects and leucocytes adhesion on renal ischemia and reperfusion injury in rats

PURPOSE: To evaluate the effects of alprostadil in an experimental model of ischemia and reperfusion injury (IRI) in rat renal tissue. METHODS: Adult male Wistar rats were randomized into three groups Vehicle-treated group(Veh), Alprostadil-treated(Al), and sham(Sh) group. Veh and Al groups had suprarenal aorta occluded for 30 minutes and reperfused for 60 minutes. Saline or 20 µg/kg of Alprostadil was intravenously infused immediately before declamping. Sh group animals underwent similar procedure without aortic occlusion. Left nephrectomy and blood sampling were performed after 60 minutes of reperfusion. Renal ICAM-1 expression and histological analysis were performed to estimate inflammatory response and tissue disarrangement. Serum biochemical markers for IRI were also measured. Kruskal-Wallis test was used to assess differences between the groups. RESULTS: There was lower expression of ICAM-1 in groups Veh and Sh. On histologically evaluation, inflammation and necrosis in the Veh group was significantly higher (grades III/IV) than Al group (Veh>Al=Sh; p = 0.025), as well as CPK levels (Veh>Al=Sh; p = 0.03). CONCLUSION: Alprostadil attenuates the immunohistochemical and histological repercussions in the renal tissue of rats submitted to a post-ischemic reperfusion with supra-renal aortic clamping. .

Philadelphia chromosome detection in chronic myeloid leukemia: Utility of phytohemagglutinin-stimulated peripheral blood culture.

Background: The conventional cytogenetic approach to demonstrate Philadelphia (Ph) chromosome at times does not yield enough number of metaphases or are of suboptimal quality. Further, the rapid molecular tests have completely pushed this simple technique into disrepute. Aims: This study aimed to evaluate usefulness of phytohemagglutinin (PHA)-stimulated peripheral blood culture for detection of Ph chromosome in chronic myeloid leukemia (CML) patients. Materials and Methods: Fifty-six patients, including 11 newly diagnosed cases of CML and 45 patients of CML on imatinib therapy showing the presence of Ph chromosome in unstimulated samples, were included in the study. Cytogenetic analysis was done on unstimulated samples, i.e. bone marrow aspirate, 24- and 48-h peripheral blood culture, and compared with PHA-stimulated 72-h peripheral blood culture. Results: The preparations from PHA-stimulated peripheral blood culture samples in all 56 patients yielded high number of good-quality metaphases. All the 11 (100%) newly diagnosed patients and 39/45 (87%) of the patients on imatinib therapy showed the presence of Ph chromosome in PHA-stimulated samples. Addition of PHA-stimulated 72-h peripheral blood culture preparation can be of use for increasing the diagnostic yield in cases of CML with suboptimal results on conventional cytogenetics from bone marrow aspirate sample.

Effect of vitamin C administration on leukocyte vitamin C level and severity of bronchial asthma

Oxidative stress mediated by reactive oxygen species is known to contribute to the inflammatory process of bronchial asthma. Reactive oxygen species are released into the bronchial tree by activated inflammatory cells. In this study, we aimed to determine the effect of vitamin C administration on leukocyte vitamin C level as well as severity of asthma. In this double blind clinical trial study we evaluated 60 patients with chronic stable asthma. The patients were divided into two groups [A and B] including 30 patients in each group. Patients in these groups were matched according to their age, weight, height, gender, BMI and drug consumption. In addition to standard asthma treatment [according to stepwise therapy in 4[th] step of bronchial asthma] in which the patients were controlled appropriately, group A received 1000 mg vitamin C daily and group B received placebo. At the baseline and after one month treatment, non-fasting blood samples were drawn for laboratory evaluations. Asthmatic patient's clinical condition was evaluated through standard pulmonary function test [PFT]. The mean [ +/- SD] leukocyte vitamin C level in group A at the baseline and after one month treatment with 1000 mg/day vitamin C, were 0.0903 [ +/- 0.0787] microg/10[8] leukocytes and 0.1400 [ +/- 0.0953] microg/10[8] leukocytes respectively [P<0.05]. The mean [ +/- SD] leukocyte vitamin C level in group B at the baseline and after one month administration of placebo, were 0.0867 [ +/- 0.0629] microg/10[8] leukocytes and 0.0805[ +/- 0.0736] microg/10[8] leukocytes respectively. The leukocyte vitamin C level in group A was higher than those of group B after one month treatment with vitamin C and placebo and the difference was statistically significant [P<0.05]. Comparing PFT [FEV[1], FVC and FEV1/FVC] in group B during the study period showed a significant increase in FEV[1] [P<0.05], while the other two parameters remained unchanged. In group A, who received 1000 mg/day vitamin C, none of the spirometry parameters changed after one month treatment, indicating no effect of vitamin C treatment in the spirometry parameters

Polymorphisms in genes involved in innate immunity and susceptibility to benzene-induced hematotoxicity

Benzene, a recognized hematotoxicant and carcinogen, can damage the human immune system. We studied the association between single nucleotide polymorphisms (SNPs) in genes involved in innate immunity and benzene hematotoxicity in a cross-sectional study of workers exposed to benzene (250 workers and 140 controls). A total of 1,236 tag SNPs in 149 gene regions of six pathways were included in the analysis. Six gene regions were significant for their association with white blood cell (WBC) counts (MBP, VCAM1, ALOX5, MPO, RAC2, and CRP) based on gene-region (P < 0.05) and SNP analyses (FDR < 0.05). VCAM1 rs3176867, ALOX5 rs7099684, and MPO rs2071409 were the three most significant SNPs. They showed similar effects on WBC subtypes, especially granulocytes, lymphocytes, and monocytes. A 3-SNP block in ALOXE3 (rs7215658, rs9892383, and rs3027208) showed a global association (omnibus P = 0.0008) with WBCs even though the three SNPs were not significant individually. Our study suggests that polymorphisms in innate immunity genes may play a role in benzene-induced hematotoxicity; however, independent replication is necessary.

Beneficial effect of Berberis buxifolia Lam, Ziziphus mistol Griseb and Prosopis alba extracts on oxidative stress induced by chloramphenicol / Efecto benéfico de extractos de Berberis buxifolia Lam, Zizyphus mistol Griseb y Prosopis alba sobre el estrés oxidativo inducido por cloramfenicol

The chemiluminescence of luminol, a measure of oxidative stress, increased immediately as a consequence of reactive oxygen species (ROS) stimulated by this antibiotic. The effect of Ch was dose dependent with maximum stimulus at 8 mg/ml (Vmax); above this concentration the cells began to reduce the production of ROS. The oxidative injury of Ch was counteracted by water extracts of Berberis buxifolia lam, Zizyphus mistol Griseb and Prosopis alba, indigenous fruits from Argentina. The relatively light units (RLU) emitted decreased immediately as a consequence of a protective effect exerted by the extracts of these fruit extracts on blood cells. The three indigenous fruit extracts reduced to a different extent the oxidative injury caused by Ch. B.buxifolia lam exhibited the highest antioxidant capacity followed by Z.mistol Griseb. Water extracts of both fruit extracts were the most effective against the oxidative stress, while P.alba presented better antioxidant capacity in the ethanolic fraction obtained. Hexane extracts showed low protective action on blood cells, with little reduction of area under curve (AUC) of RLU plotted versus time. Leukocytes remained viable in blood samples incubated for 3h with Ch and water extracts of B. buxifolia lam or Z. mistol Griseb (97.1% and 92.5% viability by Trypan blue exclusion, respectively); whereas with Ch only the cells were stressed and viability decreased to 30%. The three fruit extracts protected the viability of leukocytes in parallel with the decrease of ROS. Erythrocytes were not lysed in the presence of Ch.

Se estudió el efecto antioxidante de tres extractos de frutas autóctonas, Berberis buxifolia lam (michay), Zizyphus mistol Griseb (mistol) and Prosopis alba (algarrobo). Las células sanguíneas humanas sufrieron estrés oxidativo por acción de cloramfenicol (Ch), con un aumento inmediato de especies reactivas del oxígeno (ERO), que fue determinado por quimioluminiscencia con luminol. La respuesta fue dependiente de la dosis, con un máximo a 8 mg/ml. Los extractos de frutas autóctonas de la Argentina fueron capaces de contrarrestar el estrés generado por el antibiótico. El michay y el mistol resultaron más efectivos en la fase acuosa, y el algarrobo fue más antioxidante en extractos etílicos, mientras que las fracciones obtenidas con hexano no fueron activas. La viabilidad de los leucocitos se mantuvo elevada con Ch en presencia de extractos, entre 92.5 y 97.1%, cayendo hasta un 30% con Ch solo. Tanto los eritrocitos como los leucocitos fueron protegidos del efecto estresante por la capacidad antioxidantes de los extractos de las tres frutas investigadas, lo que podría ser importante a considerar en la dieta de niños, y pacientes en general, sometidos a Ch u otras terapias causantes de estrés oxidativo.

Melatonin treatment prevents modulation of cell-mediated immune response induced by propoxur in rats.

The effect of melatonin, a major secretory product of the pineal gland, in attenuation of propoxur (2-isopropoxy phenyl N-methyl carbamate)-induced modulation of cell-mediated immune (CMI) response was studied in rats. Male Wistar albino rats were exposed to propoxur (a widely used pesticide) orally (10 mg/kg) and/or melatonin (10 mg/kg) orally for 4 weeks. CMI was measured by delayed-type hypersensitivity (DTH), leucocyte and macrophage migration inhibition (LMI and MMI) responses and estimation of cytokines TNF-alpha and IFN-gamma levels. Rats exposed to propoxur for 4 weeks showed significant decrease in DTH, LMI and MMI responses. Propoxur also suppressed TNF-alpha and IFN-gamma production significantly. Administration of melatonin alone caused a significant increase in DTH response. Although there were no changes in the LMI and MMI response, the cytokine levels were significantly increased, as compared to control. Co-administration of melatonin along with propoxur significantly nullified the effect of the pesticide on the CMI response, except DTH and reversed levels of cytokines to near control/normal values. Thus, melatonin treatment considerably attenuated immunomodulation caused by sub-chronic treatment of propoxur in experimental animals.

Histological analysis of the association between formocresol and endotoxin in the subcutaneous tissue of mice

This study performed a histological analysis of the effect of formocresol associated to endotoxin (LPS) in the subcutaneous connective tissue of mice. Ninety mice were randomly assigned to 3 groups (n=30). Each animal received one plastic tube implant containing endotoxin solution (10 mg/mL), formocresol (original formula) or a mixture of endotoxin and formocresol. The endotoxin and formocresol groups served as controls. The periods of analysis were 7, 15 and 30 days. At each experimental period, tissue samples were collected and submitted to routine processing for histological analysis. Endotoxin and formocresol produced necrosis and chronic inflammation at 7 and 15 days. At 30 days, the endotoxin group showed no necrosis, while in the formocresol group necrosis persisted. The formocresol-endotoxin association produced necrosis and chronic inflammation in the same way as observed with formocresol at all experimental periods. In conclusion, formocresol seems not to be able to inactive the toxic effects of endotoxin in connective tissues.

O objetivo deste estudo foi avaliar histologicamente o efeito da associação do formocresol com endotoxina (LPS) em tecido conjuntivo de camundongos. Noventa camundongos foram divididos em três grupos de 30 camundongos cada. Cada camundongo recebeu um implante subcutâneo de tubo plástico contendo solução de endotoxina (10 mg/ml), formocresol (fórmula original), ou uma mistura de formocresol com endotoxina. Os grupos da endotoxina e formocresol foram considerados grupos controle. Os períodos de análise foram 7, 15 e 30 dias. Após os períodos experimentais, os tecidos foram removidos e submetidos a processamento histológico. Os resultados obtidos indicam que a endotoxina e o formocresol produzem necrose e inflamação tecidual crônica aos 7 e 15 dias e aos 30 dias o grupo da endotoxina não mostrava necrose e no grupo do formocresol a necrose persistiu. A combinação formocresol e endotoxina mostrou necrose e inflamação crônica com resultados semelhantes ao do grupo formocresol para todos os períodos experimentais. Pode-se concluir que o formocresol parece não ser capaz de inativar os efeitos tóxicos da endotoxina.

Anticoagulant induced leukoagglutination

Low leukocyte count secondary to leukocyte aggregation caused by an ethylene diamine tetra acetic acid [EDTA] occur in both benign and malignant disorders. We report a 71-year-old male patient who was admitted to the hospital with acute chest infection. Complete blood count [CBC] collected in EDTA tube and analyzed by sysmex instrument SE/9500 revealed low hemoglobin level of 9.4g/dl, white blood cell [WBC] count of 8.2x109/L and neutrophils of 4.8x109/L. Peripheral blood smear review shows multiple leukocytes aggregation one clump in each field. When we ask for another blood sample in citrate anticoagulant, the CBC showed WBC count of 11.8x109/L and neutrophils of 6.26 x 109/L. This is a case of low leukocyte count secondary to leukocyte aggregation induced by EDTA

Effect of pretreatment with chromium picolinate on haematological parameters during dengue virus infection in mice.

BACKGROUND & OBJECTIVE: Dengue virus (DV) has caused severe epidemics of dengue fever (DF) and dengue haemorrhagic fever (DHF) and is endemic all over India. We have earlier reported that exposure of mice to hexavalent chromium [Cr(VI)] compounds increased the severity of dengue virus infection. Trivalent chromium picolinate (CrP) is used worldwide as micronutrient and nutritional supplement. The present study was therefore, carried out to investigate the effects of CrP on various haematological parameters during DV infection of mice. METHODS: The Swiss Albino smice were inoculated with dengue virus (1000 LD50, intracerebrally) and fed with chromium picolinate (CrP) in drinking water (100 and 250 mg/l) for 24 wk. Peripheral blood leucocytes and other haematological parameters, and spleens were studied on days 4 and 8 after virus inoculations and the findings were compared with those given only CrP and the normal control age matched mice. RESULTS: CrP in drinking water for 24 wk had no significant effects on peripheral blood cells of mice. On the other hand, there was significant decrease in different haematological parameters following inoculation of normal mice with DV. In CrP fed mice the effects of DV infection were abolished on most of the haematological parameters. INTERPRETATION & CONCLUSION: The findings of present study showed that the adverse effects of DV infection, specially on platelets and leucocytes, were abrogated by pretreatment of mice with CrP. The therapeutic utility of CrP in viral infections including dengue needs to be studied in depth.

Immunotoxicity of carbaryl in chicken.

Effect of methyl carbonate pesticide, carbonyl, was studied on macrophage functions, lymphocyte proliferation and delayed type hypersensitivity response. Sixteen adult chicken, vaccinated against Newcaslte disease, were procured and randomly divided in two experimental groups. Chicken of group I served as control, while group II birds were given carbaryl at 20 ppm (No observable effect level, NOEL) in feed for 3 months. To measure the functional activity of phagocytic cells, nitroblue tetrazolium (NBT) reduction test was performed on peripheral blood leucocytes. Concanvalin A (Con-A) and lipopolysaccharide stimulated proliferation of T and B lymphocytes was assessed using MTT dye method. At the end of experiment, the phagocytic capacities of macrophages were significantly reduced in carbaryl treated group. Lymphocyte proliferation responses to Con-A and LPS were (23 and 28%, respectively lower) in chicken fed with carbaryl. Delayed hypersensitivity reaction to tuberculin was reduced to 77% of control values indicating inhibition of cell mediated immune response. The present study suggested of immunosuppressive effect of (NOEL dose carbaryl) in chicken.

Effects of Saussurea lapp a roots extract in ethanol on leukocyte phagocytic activity, lymphocyte proliferation and interferon - gamma [IFN-gamma]

Effects of Saussurea lappa root extracts prepared in ethanol according to the homeopathic principles were ssessed on leukocyte phagocytic activity, lymphocyte transformation and mitogen-induced interferon-gamma [INF-gamma] in the cultures of peripheral blood mononuclear cells of goats [PBMC] in vitro. Leukocyte phagocytic activity was measured by flow cytometry, lymphocyte proliferation by MTT and IFN-gamma level in cell culture supernatants was determined by ELISA. The results obtained demonstrated that all tests dilutions [D4, D6, D8] of Saussurea lappa in ethanol have exerted a stimulating effect on leukocyte phagocytic activity in dose-dependent manner. A 10 micro liters dose of Saussurea lappa of each dilution markedly enhanced phagocytic activity, while other does tested made only a feeble stimulating effect. The increase with 10 micro liter dose were found significantly [p<0.01] different between each dilution, maximal stimulationwas observed by D8 dilution. Different does [10 micro liters, 2 micro liters, 0.5 micro liters] of all test dilutions [D4, D6, D8] of Saussurea lappa in sterile 0.9% NaCl solution inhibited lymphocyte proliferation. Maximal inhibitory effect was observed with the 2 micro liter dose. Similarly, Saussurea lappa suppressed the secretion of IFN-gamma by mitoren-activated [PHA; 2.5 micro g/ml] of peripheral mononuclear cells in dose-dependent manner. In conclusion these findings suggest that enhanced leukocyte phagocytic activity may be helpful to clear the soluble immune complexes produced during a sustained immune response against self antigens which causes chronic inflammatory injury of tissue. On the other hand, inhibition of lymphocyte proliferation and IFN-gamma by Saussurea lappa may contribute to suppress immune-mediated inflammatory reactions possibly through a cell-mediated cytokine pathway. Thus it is conceivable that ethanolic extracts of Saussurea lappa roots in homeopathetic dilutions may be considered as a potential candidate for therapeutic support in autoimmune and chronic inflammatory disorders

Phytotherapeutic effects of Echinacea purpurea in gamma-irradiated mice

Echinacea (E.) purpurea herb is commonly known as the purple coneflower, red sunflower and rudbeckia. In this paper, we report the curative efficacy of an Echinacea extract in gamma-irradiated mice. E. purpurea was given to male mice that were divided into five groups (control, treated, irradiated, treated before irradiation & treated after irradiation) at a dose of 30 mg/kg body weight for 2 weeks before and after irradiation with 3 Gy of gamma-rays. The results reflected the detrimental reduction effects of gamma-rays on peripheral blood hemoglobin and the levels of red blood cells, differential white blood cells, and bone marrow cells. The thiobarbituric acid-reactive substances (TBARs) level, Superoxide dismutase (SOD) and glutathione peroxidase (GSPx) activities and DNA fragmentation were also investigated. FT-Raman spectroscopy was used to explore the structural changes in liver tissues. Significant changes were observed in the microenvironment of the major constituents, including tyrosine and protein secondary structures. E. purpurea administration significantly ameliorated all estimated parameters. The radio-protection effectiveness was similar to the radio-recovery curativeness in comparison to the control group in most of the tested parameters. The radio-protection efficiency was greater than the radio-recovery in hemoglobin level during the first two weeks, in lymphoid cell count and TBARs level at the fourth week and in SOD activity during the first two weeks, as compared to the levels of these parameters in the control group.

Effects of simvastatin in abdominal sepsis in rats

PURPOSE: 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-alpha, IL-1beta 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-alpha, IL-1beta 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.

OBJETIVO: As estatinas são agentes reconhecidamente hipolipemiantes. Vários estudos têm revelado que eles têm ações pleiotrópicas, como antiinflamatória e imunomoduladora. Tentando-se entender o papel antiinflamatório da sinvastatina na sepse, foram analisados os níveis de citocinas pró-inflamatórias e contagem de leucócitos em modelo de sepse abdominal por ligadura e punção do ceco (LPC) em ratos. MÉTODOS: Foram utilizados 28 ratos Wistar pesando 285±12g, assim divididos: grupo sepse (n=14), submetidos a LPC e grupo sham (n=14), submetidos a laparotomia e manipulação suave do ceco. No grupo LPC/sinvastatina (n=7) os ratos receberam 10mg/kg de sinvastatina via oral 18 e 2 horas antes da LPC e no grupo LPC/salina (n=7) os ratos receberam injeção oral de solução salina 0,9 por cento. Os animais dos grupos sham/sinvastatina (n=7) e sham/salina (n=7) receberam o mesmo tratamento. Dosagem de TNF-alfa, IL-1beta e IL-6 por ELISA e contagem de leucócitos totais, neutrófilos, linfócitos e eosinófilos foram realizadas em todos os animais. Análise estatística foi feita pelo ANOVA e teste de Tukey, com significância p<0,05. RESULTADOS: Ficou demonstrado que as dosagens de TNF-alfa, IL-1beta e IL-6 atingiram valores de 364,8±42pg/ml; 46,3±18pg/ml e 28,4±13pg/ml no grupo submetido à sepse e tratados com sinvastatina, significantemente mais baixos do que no grupo sepse não tratados (778,5±86pg/ml; 176,9±46pg/ml; 133,6±21 pg/ml, respectivamente). O mesmo ocorreu na contagem de leucócitos totais e neutrófilos. CONCLUSÃO: A sinvastatina mostrou ação anti-inflamatória em ratos Wistar, diminuiu níveis de citocinas e leucócitos, sugerindo uso potencial na prevenção ou atenuação dos efeitos da sepse abdominal.

Indole ring oxidation by activated leukocytes prevents the production of hypochlorous acid

Hypochlorous acid (HOCl) released by activated leukocytes has been implicated in the tissue damage that characterizes chronic inflammatory diseases. In this investigation, 14 indole derivatives, including metabolites such as melatonin, tryptophan and indole-3-acetic acid, were screened for their ability to inhibit the generation of this endogenous oxidant by stimulated leukocytes. The release of HOCl was measured by the production of taurine-chloramine when the leukocytes (2 x 10(6) cells/mL) were incubated at 37°C in 10 mM phosphate-buffered saline, pH 7.4, for 30 min with 5 mM taurine and stimulated with 100 nM phorbol-12-myristate acetate. Irrespective of the group substituted in the indole ring, all the compounds tested including indole, 2-methylindole, 3-methylindole, 2,3-dimethylindole, 2,5-dimethylindole, 2-phenylindole, 5-methoxyindole, 6-methoxyindole, 5-methoxy-2-methylindole, melatonin, tryptophan, indole-3-acetic acid, 5-methoxy-2-methyl-3-indole-acetic acid, and indomethacin (10 æM) inhibited the chlorinating activity of myeloperoxidase (MPO) in the 23-72 percent range. The compounds 3-methylindole and indole-3-acetic acid were chosen as representative of indole derivatives in a dose-response study using purified MPO. The IC50 obtained were 0.10 ± 0.03 and 5.0 ± 1.0 æM (N = 13), respectively. These compounds did not affect the peroxidation activity of MPO or the production of superoxide anion by stimulated leukocytes. By following the spectral change of MPO during the enzyme turnover, the inhibition of HOCl production can be explained on the basis of the accumulation of the redox form compound-II (MPO-II), which is an inactive chlorinating species. These results show that indole derivatives are effective and selective inhibitors of MPO-chlorinating activity.

Evaluation of mercury toxicity by some cytological indices in leucocyte cultures.

The genotoxicity induced by different levels of inorganic mercury was evaluated by chromosome metaphase analysis in human leucocytes, treated in vitro for 72 hr. Mitotic index gradually decreased with an increase in concentration of mercury but the reverse phenomenon was observed with respect to chromosomal aberration due to its probable interaction with protein and DNA.

Effects of dengue virus infection on peripheral blood cells of mice exposed to hexavalent chromium with drinking water.

BACKGROUND & OBJECTIVE: The occupational and non-occupational exposure to hexavalent chromium Cr (VI) is common. The effect of chromium compromises the immune response of the host. Dengue virus (DV) infection causes various changes in the peripheral blood cells. It is, therefore, possible that the chromium toxicity may affect the disease process during DV infection. The present study aims to study the effects of dengue virus infection on peripheral blood cells of mice fed Cr (VI) with drinking water. METHODS: One group of mice was given ad libitum drinking water containing Cr (VI) and the other group used as the normal control mice was given plain water to drink. At the 3, 6 and 9 wk of Cr (VI) drinking, a set of mice from each group was inoculated intracerebrally (ic) with DV and studied at the 4th and 8th day post inoculation. RESULTS: It was observed that Cr (VI) drinking led to reduction in lymphocytes, haemoglobin and the haematocrit values while the granulocyte, monocyte and platelet counts were increased. On the other hand, most of the parameters were decreased following inoculation of normal mice with DV. In Cr (VI)-fed mice the effects of DV infection were minimal. The most significant finding of these experiments was that the reduction in platelet counts following inoculation with DV was markedly less in Cr (VI)-fed mice than that in DV-inoculated normal control mice. INTERPRETATION & CONCLUSION: Cr(VI) compounds have been declared as a potent occupational carcinogen. On the contrary, Cr(III) salts such as chromium polynicotinate, chromium chloride and chromium picolinate, are used as micronutrients and nutritional supplements, and have been shown to exhibit health benefits in animals and humans. Whether therapeutic doses of chromium (III) compounds may be able to prevent the DV-induced fall in platelet counts, needs to be investigated.

An overview of the effects of annexin 1 on cells involved in the inflammatory process

The concept of anti-inflammation is currently evolving with the definition of several endogenous inhibitory circuits that are important in the control of the host inflammatory response. Here we focus on one of these pathways, the annexin 1 (ANXA1) system. Originally identified as a 37 kDa glucocorticoid-inducible protein, ANXA1 has emerged over the last decade as an important endogenous modulator of inflammation. We review the pharmacological effects of ANXA1 on cell types involved in inflammation, from blood-borne leukocytes to resident cells. This review reveals that there is scope for more research, since most of the studies have so far focused on the effects of the protein and its peptido-mimetics on neutrophil recruitment and activation. However, many other cells central to inflammation, e.g. endothelial cells or mast cells, also express ANXA1: it is foreseen that a better definition of the role(s) of the endogenous protein in these cells will open the way to further pharmacological studies. We propose that a more systematic analysis of ANXA1 physio-pharmacology in cells involved in the host inflammatory reaction could aid in the design of novel anti-inflammatory therapeutics based on this endogenous mediator.