Comparative evaluation of plant extract effects on peritoneal, medullary and J774 cells. G8 macrophages.

The use of medicinal plants as raw material for extracts production and pure substances isolation and subsequence development of new drugs represents a constantly growing area. However, some stages are indispensable before pharmacologically evaluating natural products such as medicines. Toxicity tests in mammalian cells are essential to initiate new drugs development or verify the substance's biocompatibility. Thus, we verified the toxicity of crude extracts and fractions with different polarities obtained from the leaves and stems of eight plant species. The toxic effect was evaluated on macrophages obtained from the bone marrow and peritoneal cavity of a Swiss webster mouse and J774 macrophages. G8 cell lineage. These macrophages were cultured in a 96-well plate, and the compounds were added at a concentration of 100 µg/mL for 24 hours. After this time, the supernatant was removed. The toxicity was evaluated for lactate dehydrogenase (LDH) release assay and the resazurin assay, which uses an indicator dye to measure oxidation-reduction reactions. The results showed a difference in the percentage of toxicity when comparing the same extract in different types of macrophages. This outcome indicates that these cells from different origins may exhibit different responses when exposed to the same natural compounds.

Comparative evaluation of plant extract effects on peritoneal, medullary and J774 cells G8 macrophages / Avaliação comparativa do efeito de extratos de plantas em macrófagos peritoneais, medulares e da linhagem J774. G8

The use of medicinal plants as raw material for extracts production and pure substances isolation and subsequence development of new drugs represents a constantly growing area. However, some stages are indispensable before pharmacologically evaluating natural products such as medicines. Toxicity tests in mammalian cells are essential to initiate new drugs development or verify the substance's biocompatibility. Thus, we verified the toxicity of crude extracts and fractions with different polarities obtained from the leaves and stems of eight plant species. The toxic effect was evaluated on macrophages obtained from the bone marrow and peritoneal cavity of a Swiss webster mouse and J774 macrophages. G8 cell lineage. These macrophages were cultured in a 96-well plate, and the compounds were added at a concentration of 100 µg/mL for 24 hours. After this time, the supernatant was removed. The toxicity was evaluated for lactate dehydrogenase (LDH) release assay and the resazurin assay, which uses an indicator dye to measure oxidation-reduction reactions. The results showed a difference in the percentage of toxicity when comparing the same extract in different types of macrophages. This outcome indicates that these cells from different origins may exhibit different responses when exposed to the same natural compounds.

A utilização de plantas medicinais como matéria-prima para a produção de extratos e isolamento de substâncias puras para o desenvolvimento de novos fármacos representa uma área em constante crescimento. No entanto, existem processos a serem realizados antes de avaliar farmacologicamente produtos naturais como medicamentos. Os testes de toxicidade em células de mamíferos são fundamentais para iniciar o desenvolvimento de novas drogas ou verificar a biocompatibilidade de substâncias. Assim, verificamos a toxicidade de extratos brutos e frações com diferentes polaridades obtidos de folhas e caule de oito espécies de planta. Para comparar o efeito tóxico, os testes foram realizados em macrófagos obtidos da medula óssea e cavidade do peritônio de camundongo Swiss webster, bem como no macrófago da linhagem celular J774.G8. Esses macrófagos foram cultivados em placa de 96 poços e os compostos adicionados na concentração de 100 µg/mL por 24 horas. Após esse período o sobrenadante foi removido. A toxicidade foi avaliada pelos ensaios de detecção da enzima lactato-desidrogenase (LDH) e pelo ensaio de resazurina, que usa um corante indicador para medir as reações de oxidação-redução. Os resultados mostraram uma diferença na porcentagem de toxicidade quando comparamos o mesmo extrato em diferentes tipos de macrófagos. Este resultado indica que essas células de várias origens podem exibir respostas distintas quando expostas aos mesmos compostos naturais.

Effects of ractopamine administration and castration method on the response to preslaughter stress and carcass and meat quality in pigs of two Piétrain genotypes.

The objective of this study was to evaluate the effects of ractopamine supplementation, castration method, and their interaction on the behavioral and physiological response to preslaughter stress and carcass and meat quality of 2 Piétrain genotypes. A total of 1,488 male pigs (115 ± 5 kg BW) were distributed according to a 2 × 2 × 2 factorial arrangement of treatments. The first factor was ractopamine supplementation with 2 groups of pigs (376 and 380 pigs each) receiving 7.5 mg/kg of ractopamine (RAC) or not (NRAC) in their diet during the last 28 d of the finishing period. The second factor was castration method, with 744 surgical castrates (SC) and 744 immunized males (IM), and the third factor was the genotype with 2 crossbreeds containing 50% (genotype A, GA; n = 744) or 25% (genotype B, GB; n = 744) Piétrain genetics. Surgical castration took place at 2 d of age, whereas immunization against gonadotropin-releasing factor (GnRF) was performed through 2 subcutaneous injections of GnRF analog (Improvest, 2 mL) at 10 and 4 wk before slaughter. At loading more vocal stimulation was needed by the handler to drive GB pigs forward through the farm alley (P = 0.01) and RAC-fed GB pigs through the ramp (P = 0.02). Feeding RAC to IM increased the number of fights in lairage compared with SC (P = 0.03). Feeding RAC shortened fighting bouts compared with NRAC pigs (P = 0.05). The SC-GA pigs showed a greater gastrointestinal tract temperature during unloading (P = 0.05) and lairage time (P = 0.03). Blood creatine kinase (CK) concentrations were greater (P = 0.04) in SC compared with IM, and no difference was found in the concentrations of stress hormones in urine collected postmortem. Dressing yield was greater (P = 0.01) in RAC and SC-GB pigs. Carcasses from RAC pigs and IM were leaner than those from NRAC and SC pigs (P < 0.001 and P = 0.002, respectively). Feeding RAC to IM increased drip loss in the LM (P = 0.05). Warner-Bratzler shear force values were slightly greater in the LM from RAC-GB pigs and from IM compared with SC (P = 0.01 and P < 0.001, respectively) and in the semimembranosus muscle of RAC pigs (P = 0.006). In conclusion, immunization against GnRF more than the use of Piétrain genotypes appears to be a viable alternative to the use of ractopamine, as it seems to promote production of lean carcasses without compromising animal welfare and pork quality.

Comparative phenotypic profile of subpopulations of peripheral blood leukocytes in European (Bos taurus taurus) and Zebu cattle (Bos taurus indicus).

Differences in cellular and humoral immunity in Zebu (Bos taurus indicus) and European (B. taurus taurus) cattle breeds, which may be related to differences in resistance or susceptibility to infectious or parasitic diseases, are largely unknown. This study aimed to perform a comparative analysis of innate and adaptive immunity of European (including Holstein, Brown Swiss, and Hereford) and Zebu (including Gir, Nelore, and Guzera) breeds, by assessing their peripheral blood leukocyte profiles (i.e., monocytes, eosinophils, and lymphocytes, including CD4(+) and CD8(+) T cells, and CD21(+) B cells). Higher frequencies of cells involved in innate immunity were observed in Zebu breeds, particularly monocytes and non-T and non-B cells (13.37 ± 0.9058 and 37.67 ± 1.55, respectively). This finding may contribute to the increased resistance of B. taurus indicus to certain infectious and parasitic diseases. Considering other leukocyte populations in the peripheral blood, among-breed variation was greater than differences between the two subspecies. This study will serve as a basis for further investigations regarding comparative immunology and resistance to infectious and parasitic diseases of cattle.

Chemical composition and antibacterial activity of Brazilian propolis essential oil

The present study aimed at investigating the chemical composition of essential oil extracted from Brazilian propolis and the susceptibility of Staphylococcus aureus, Staphylococcus epidermides, Streptococcus pyogenes and Escherichia coli to this substance. The essential oil was obtained by steam distillation of propolis and examined by gas chromatography/mass spectrometry (GC/MS). In addition, the agar diffusion method using filter paper disks was employed. Antibacterial activity was measured as equivalent diameters of inhibition zones (in millimeters) after incubation at 37ºC for 24 hours. From the 26 identified constituents, Beta-caryophyllene (12.7 percent), acetophenone (12.3 percent) and Beta-farnesene (9.2 percent) were found to be major components. New components, namely linalool, methyl hydrocinnamate, ethyl hydrocinnamate, alfa-ylangene, gama-elemene and valencene, are reported for the first time to be present in propolis essential oil. This oil also exhibited antibacterial activity.

Chemical composition and antibacterial activity of Brazilian propolis essential oil

The present study aimed at investigating the chemical composition of essential oil extracted from Brazilian propolis and the susceptibility of Staphylococcus aureus, Staphylococcus epidermides, Streptococcus pyogenes and Escherichia coli to this substance. The essential oil was obtained by steam distillation of propolis and examined by gas chromatography/mass spectrometry (GC/MS). In addition, the agar diffusion method using filter paper disks was employed. Antibacterial activity was measured as equivalent diameters of inhibition zones (in millimeters) after incubation at 37ºC for 24 hours. From the 26 identified constituents, Beta-caryophyllene (12.7 percent), acetophenone (12.3 percent) and Beta-farnesene (9.2 percent) were found to be major components. New components, namely linalool, methyl hydrocinnamate, ethyl hydrocinnamate, alfa-ylangene, gama-elemene and valencene, are reported for the first time to be present in propolis essential oil. This oil also exhibited antibacterial activity.(AU)

Results of en bloc renal transplants of pediatric deceased donors into adult recipients.

UNLABELLED: Various strategies have evolved to expand the donor pool due to the extreme shortage of organs. Herein we reviewed our experience with en bloc pediatric kidney transplantation since 1998. METHODS: From January 1998 to December 2004, nine adult patients underwent kidney transplantation using en bloc kidneys from donors <5 years old (range, 1 to 4). The mean age of the recipients was 45.1 years (range, 34 to 57). RESULTS: In recipients of en bloc pediatric transplantation, cold ischemia time ranged from 14 to 26.2 hours (mean, 21.3 hours). Mean serum creatinine at 3, 6, and 12 months after transplantation was 1.53 +/- 0.57, 1.27 +/- 0.27, and 1.15 +/- 0.26 mg/dL compared with 1.93 +/- 1.35, 1.81 +/- 1.17, and 1.73 +/- 0.85 (P = .08) in recipients of single kidneys from ideal cadaveric donors (UNOS criteria, n = 368). Patient and graft survival at 1 year were 88.8% compared with 91.2% and 85% with ideal donors (P = NS), respectively. Three cases required additional surgery. There was one death due to a cerebral vascular accident. CONCLUSION: The present study confirmed the excellent results achieved with transplantation using en bloc kidneys from young donors.

Effects of homeopathic treatment on pruritus of haemodialysis patients: a randomised placebo-controlled double-blind trial.

Pruritus is a frequent and difficult to treat problem in haemodialysis. This double-blind placebo-controlled randomised clinical trial assessed the role of homeopathic treatment in this situation. The code was held by the pharmacist who dispensed the medications. Pruritus was evaluated using a previously published scale. Only patients with initial values above 25% of maximum pruritus score were entered. Data were analysed after partial code break, separating the two groups of patients, but with no awareness of which one received verum or placebo. Patients were classified as responders if they had >50% reduction of pruritus score. Twenty-eight patients (16M/12F, 51 +/- 11 years of age) were entered and 20 (12M/8F, 52 +/- 8 years of age) remained for final analysis: 11 in the verum group, 9 in placebo. At entry, the mean pruritus score was 65 +/- 25% for the treated patients and 70 +/- 27% for placebo. After 15, 30, 45, and 60 days of follow-up, pruritus score were respectively: 46 +/- 29, 41 +/- 30, 42 +/- 29, and 38 +/- 33 for the treated patients and 61 +/- 29, 67 +/- 31, 64 +/- 35, and 57 +/- 39 for placebo. Reduction was statistically significant (P<0.05) at every point of observation. According to the patients' own assessment, at the end of the study period, the homeopathic treatment reduced the pruritus score by approximately 49%. Responders were more frequent in the treated group with statistical significance at 30 days (0% vs 45%, P=0.038). Homeopathic treatment may represent a worthwhile alternative to relieve pruritus in patients undergoing haemodialysis.

Effects of homeopathic treatment on pruritus of haemodialysis patients: a randomised placebo-controlled double-blid trial

Pruritus is a frequent and difficult to treat problem in haemodialysis. This double-blid placebo-controlled randomised clinical trial assessed the role of homeopathic treatment in this situation. The code was held by the pharmacist... (AU)

Safety and bactericidal activity of rifalazil in patients with pulmonary tuberculosis.

Rifalazil, also known as KRM-1648 or benzoxazinorifamycin, is a new semisynthetic rifamycin with a long half-life of approximately 60 h. Rifalazil has potent bactericidal activity against Mycobacterium tuberculosis in vitro and in animal models of tuberculosis (TB). Prior studies in healthy volunteers showed that once-weekly doses of 25 to 50 mg of rifalazil were well tolerated. In this randomized, open-label, active-controlled phase II clinical trial, 65 subjects with sputum smear-positive pulmonary TB received one of the following regimens for the first 2 weeks of therapy: 16 subjects received isoniazid (INH) (5 mg/kg of body weight) daily; 16 received INH (5 mg/kg) and rifampin (10 mg/kg) daily; 17 received INH (5 mg/kg) daily plus 10 mg of rifalazil once weekly; and 16 received INH (5 mg/kg) daily and 25 mg of rifalazil once weekly. All subjects were then put on 6 months of standard TB therapy. Pretreatment and day 15 sputum CFU of M. tuberculosis were measured to assess the bactericidal activity of each regimen. The number of drug-related adverse experiences was low and not significantly different among treatment arms. A transient decrease in absolute neutrophil count to less than 2,000 cells/mm(3) was detected in 10 to 20% of patients in the rifalazil- and rifampin-containing treatment arms without clinical consequences. Decreases in CFU counts were comparable among the four treatment arms; however, the CFU results were statistically inconclusive due to the variability in the control arms. Acquired drug resistance did not occur in any patient. Studies focused on determining a maximum tolerated dose will help elucidate the full anti-TB effect of rifalazil.

Infection and disease among household contacts of patients with multidrug-resistant tuberculosis.

SETTING: Urban public teaching and referral hospital in Espirito Santo, Brazil. OBJECTIVE: To assess whether rates of infection and progression to active tuberculosis (TB) differed between household contacts of patients with multidrug-resistant (MDR) and drug susceptible (DS) pulmonary tuberculosis. DESIGN: Household contacts were assessed for evidence of TB infection and disease by purified protein derivative (PPD) skin testing, physical examination, chest X-ray, and sputum smear and culture. RESULTS: Among 133 close contacts of patients with MDR-TB, 44% were PPD-positive (> or =10 mm) compared to 37% of 231 contacts of the DS-TB cases (P = 0.18, chi2 test, OR 1.2, 95%CI 0.8-2). In a multivariate logistic regression analysis, after allowance for between-household variation in PPD responses, PPD positivity among household contacts of patients with MDR-TB remained comparable to PPD positivity in contacts of patients with DS-TB (OR 2.1, 95%CI 0.7-6.5). Respectively six (4%) and 11 (4%) contacts of the MDR- and DS-TB cases were found to have active TB at the time of initial evaluation or during follow-up (P = 0.78, chi2 test). Five of six contacts of MDR-TB cases and nine of nine contacts of DS-TB cases who developed TB, and for whom drug susceptibility test results were available, had the same bacterial susceptibility profiles as their index cases. DNA fingerprinting analysis of Mycobacterium tuberculosis isolates was identical between household contacts with active TB and the index MDR or DS-TB case for all 14 pairs compared. CONCLUSION: Our data suggest that the prevalence of tuberculous infection and progression to active TB among household contacts exposed to DS and MDR-TB cases is comparable, despite a longer duration of exposure of contacts to the index case in patients with MDR-TB.