Does the tumour microenvironment alter tumorigenesis and clinical response in transmissible venereal tumour in dogs?

The canine transmissible venereal tumour (CTVT) is a transmissible cancer that is spread naturally between dogs, with the ability to develop and evade the immune system, despite strict immune surveillance of the host. Furthermore, molecular signalling between cells of the immune system and the tumour microenvironment appear to influence the behaviour and development of the tumour. Thus, this study aimed to quantify the expression of genes related to the immune system such as IL‐6, IFN‐γ, and TGF‐β, as well as angiogenic factors (VEGF, CXCR4), in CTVT cells in vivo and in vitro (primary culture), correlating with the clinical response of the animals treated with vincristine. As expected, the most prevalent subtype was plasmacytoid cells, although lymphocytic cells were also found, indicating the possibility of polyclonality. When we compared the gene expressions of IFN‐γ and IL‐6, we mostly found low expression, concluding that MHC expression was probably not occurring in tumour cells, and no activation of immune cells to eliminate the tumour. The TGF‐β gene was normal in the majority of animals but demonstrated decreased expression in vincristine resistant animals, leading to the hypothesis that the concentration of tumour‐derived TGF‐β was affecting and even suppressing the real TGF‐β expression, favouring tumour proliferation and progression in these cases. VEGF expression was extremely high, demonstrating its angiogenic role in tumour growth, while CXCR4 was decreased, possibly because of CTVT’s low metastatic potential. Thus, we concluded that the tumour microenvironment, together with the immune system of the host, influences CTVT, presumably altering its tumorigenesis and the animal’s clinical response to treatment.     

Haematological and histopathological findings after ovariectomy in Trypanosoma cruzi infected mice

The aim of this study was to investigate the influences of ovariectomy on histopathological and hematological parameters during the course of Trypanosoma cruzi infection. Hematological and immunological homeostasis is influenced by gonadal steroid hormones. Ovariectomy exerts profound influences on parasitic diseases including T. cruzi infection through modulation of the host's immune response. Three groups of female Mus musculus were infected with 4000 blood trypomastigotes of the Y strain of T. cruzi. One group was subjected to ovariectomy, another to simulated surgery before the infection, and a third group of unoperated animals were used as controls. Marked differences were detected in the responses of blood and tissue parasites. On day 9, post-infection parasitism was significantly higher in ovariectomized animals (P<0.05). These results were confirmed by histopathological studies, in which ovariectomized animals displayed hearts with higher number of amastigote burdens, increased inflammatory infiltrate, enhanced tissue fibers disorganization and decreased lytic antibody percentage, when compared to their counterparts. On day 9 the hematological changes were more apparent, with a decrease in erythrocytes, platelets and leucocytes for ovariectomized infected animals. Simulated surgery, as a stressful agent, did not cause any imbalance in parasitism or in the hemogram profile. The results confirm the importance of the female steroids in resistance against T. cruzi infection.     

Evaluation of transformation growth factor beta1, interleukin-10, and interferon-gamma in male symptomatic and asymptomatic dogs naturally infected by Leishmania (Leishmania) chagasi

The aims of this study were to evaluate the immunomodulatory role of TGF-β₁, IL-10, and INF-γ in spleen and liver extracts and supernatant cultures of white spleen cells from male symptomatic and asymptomatic dogs, naturally infected by Leishmania (Leishmania) chagasi. Thirty dogs from Araçatuba, São Paulo, Brazil, an endemic leishmaniosis area, were selected by positive ELISA serological reaction for Leishmania sp. and divided into two groups: asymptomatic (n = 15) and symptomatic (n = 15) consisting of animals with at least three characteristic signs (fever, dermatitis, lymphoadenopathy, onychogryphosis, weight loss, cachexia, locomotion problems, conjunctivitis, epistaxis, hepatosplenomegaly, edema, and apathy). After euthanasia, spleen and liver fragments were collected for ex vivo quantification of TGF-β₁, IL-10, and INF-γ. Naturally active in vitro produced TGF-β₁ was also evaluated in spleen cell culture supernatant. Spleen and liver extract of asymptomatic dogs had higher mean TGF-β₁ levels than symptomatic dogs. High concentrations of IL-10 were found in spleen, and mainly in liver extract of both groups. Higher INF-γ concentrations were found in spleen extracts of symptomatic dogs, and in liver extracts of asymptomatic dogs. Extract of this cytokine was lower in spleen extract. Although INF-γ is being produced in canine infection, mean levels of TGF-β₁ and IL-10 from spleen and liver extracts were quantitatively much higher; suggesting that immune response in both asymptomatic and symptomatic dogs was predominantly type Th2.     

An investigation of Leishmania spp. in Didelphis spp. from urban and peri-urban areas in Bauru (São Paulo, Brazil)

Blood and bone marrow samples were taken from 112 Didelphis spp., collected between March 2005 and February 2006, from urban and peri-urban areas of Bauru, São Paulo State, Brazil, to evaluate the hypothesis that these animals might constitute a reservoir of Leishmania spp. Anti-Leishmania ssp. antibodies were screened in the serum samples using an enzyme-linked immuno-sorbent assay (ELISA) and the polymerase chain reaction (PCR). PCR was performed on fragments of DNA samples from Leishmania spp. using primers 13A and 13B, and showed a positive outcome in 91.6% of the 112 samples tested. Of the 107 samples analyzed by ELISA, 71% were positive. Evidence of epidemiological risk factors such as a circulating parasite and freely moving vectors suggests that Didelphis spp. may participate in the transmission cycle of Leishmania spp. in Bauru.     

A dose-response study relating the concentration of carotenoid pigments in blood and reflectance spectrum characteristics of fat to carotenoid intake level in sheep

This study was conducted to describe the dose-response curve relating the concentration of carotenoid pigments in plasma and reflectance spectrum characteristics of fat to the carotenoid intake level in sheep, and to investigate the extent to which incorporation of dehydrated alfalfa in the diet affects the reliability of the discrimination between concentrate-fed and pasture-fed lambs based on these measurements. In Exp. 1, 6 treatments were compared in individually penned lambs: feeding 0, 250, 500, 750, 1,000, or 1,250 g/d of dehydrated alfalfa for 60 d before slaughter. Each treatment (T0 to T1,250) consisted of 8 male Romanov x Berrichon lambs with an initial average BW of 24.8 kg (SD 2.6). All lambs received straw for ad libitum intake and T0 to T1,000 lambs received a concentrate free of green vegetative matter in amounts to produce similar ADG in all treatments. In Exp. 2, 33 male Romanov x Berrichon lambs grazed a natural pasture maintained in a leafy green vegetative stage for at least 59 d before slaughter. Initial BW when turning out to pasture was 14.2 kg (SD 2.3). Plasma carotenoid concentration was measured at slaughter by spectrophotometry. Reflectance spectrum, lightness, redness, and yellowness were measured after 24 h of shrinkage in subcutaneous caudal and perirenal fat. The spectra were translated to 0 reflectance at 510 nm, and the integral of the translated spectrum was calculated between 450 and 510 nm (i.e., the range of light absorption by carotenoids). Reflectance measurement was replicated 5 times, from which we calculated the absolute value of the mean integral (AVMI). In Exp. 1, plasma carotenoid concentration at slaughter (PCCS) increased linearly with mean daily carotenoid intake (P < 0.01). Both subcutaneous caudal and perirenal fat AVMI increased linearly (P < 0.01) with mean daily carotenoid intake and PCCS, the slopes of the regressions being greater for perirenal than for subcutaneous caudal fat. The mean PCCS was greater for lambs of Exp. 2 than for lambs on any treatment of Exp. 1 (P < 0.01). We established the dose-response curves relating PCCS and AVMI of subcutaneous and perirenal fat to carotenoid intake level. The combined use of PCCS and of perirenal fat AVMI enabled discrimination of pasture-fed lambs of Exp. 2 from the lambs of Exp. 1 that received up to 500 g/d of dehydrated alfalfa.     

Immobilization of the pesticide 2,4‐dichlorophenoxyacetic acid on a silica gel surface

A route for the immobilization of 2,4‐dichlorophenoxyacetic acid (2,4‐D) on silica gel has been developed. In the first step the precursor was prepared by reacting the silylating agent 3‐(trimethoxysilyl)propylamine with silica gel. Nitrogen analysis of this anchored compound showed the presence of 1.16 mmol of amine groups per gram of support. The herbicide was covalently bonded to the amine groups previously anchored onto silica gel. Infrared, [¹³C] and [²⁹Si] NMR spectra supported the reaction between the nitrogen of the amine group of the anchored silica with the carbon on the para‐position of the aromatic ring of 2,4‐D. The reaction yield, 90.3%, was confirmed through elemental analysis. © 2000 Society of Chemical Industry     

Trypanocidal Action of (?)-Elatol Involves an Oxidative Stress Triggered by Mitochondria Dysfunction

Natural compounds have shown good potential for the discovery of new chemotherapeutics for the treatment of Chagas’ disease. Recently, our group reported the effective trypanocidal activity of (−)-elatol, extracted from the red macroalgae Laurencia dendroidea present in the Brazilian coast against Trypanosoma cruzi. However, the mechanism of action of this compound has remained unclear. There are only hypotheses concerning its action on mitochondrial function. Here, we further investigated the mechanisms of action of (−)-elatol on trypomastigotes of T. cruzi. For this, we evaluated some biochemical alterations in trypomastigotes treated with (−)-elatol. Our results show that (−)-elatol induced depolarization of the mitochondrial membrane, an increase in the formation of mitochondrial superoxide anion and loss of cell membrane and DNA integrity. Additionally, (−)-elatol induced formation of autophagic vacuoles and a decrease in cell volume. All together, these results suggest that the trypanocidal action of (−)-elatol involves multiple events and mitochondria might be the initial target organelle. Our hypothesis is that the mitochondrial dysfunction leads to an increase of ROS production through the electron transport chain, which affects cell membrane and DNA integrity leading to different types of parasite death.     

Using Limestone to Improve Soil Fertility and Growth of Mango (Mangifera Indica L.)

Soil acidity is one of the most important factors limiting crop production. The objective of this work was to evaluate the effects of limestone application on the soil chemical properties, nutrition and yield of mango plants in an orchard under implementation. The design was randomized blocks, with five limestone doses (0; 2; 4; 6 and 8 t ha^?1) and four replications. Soil chemical analyses were performed (at 12, 24, 36 and 48 months after the experiment implementation) in the layers 0–20; 20–40 and 40–60 cm deep. Nutrition status and yield were assessed during the first and second crop seasons. The highest fruit yield was associated with the application of 3.9 and 3.8 t ha^?1 of limestone in the 2008/09 and 2009/10 seasons, respectively, that is, the dose recommended by the literature for raising base saturation to 80%, as a function of the fertility conditions of the soil initially obtained.