Comparison of the chemical composition of extracts from Scutellaria lateriflora using accelerated solvent extraction and supercritical fluid extraction versus standard hot water or 70% ethanol extraction

The aqueous extract of American skullcap (Scutellaria lateriflora L. (S. lateriflora), Lamiaceae) has been traditionally used by North American Indians as a nerve tonic and for its sedative and diuretic properties. Recent reports stated that flavonoids and possibly amino acids are responsible for the anxiolytic activity. As a part of our search for environmentally friendly solvents to extract the active components from medicinal plants, we used S. lateriflora in a comparison of accelerated solvent extraction (ASE) using water, and supercritical fluid extraction (SFE) using CO2 and 10% EtOH as modifier, at different temperatures. Flavonoids and amino acids were quantified by HPLC-UV and HPLC-MS, respectively. The flavonoid content was compared with conventional extraction methods (hot water extraction and 70% ethanol). The use of ASE at 85 degrees C with water as solvent gave the best results for flavonoid glycosides and amino acids, whereas SFE gave higher yields of flavonoid aglycones. However, the results obtained for total flavonoids were not significatively superior to hot water extraction or 70% aqueous EtOH extract.     

Determination of anthocyanins from camu-camu (Myrciaria dubia) by HPLC-PDA, HPLC-MS, and NMR

Camu-camu [Myrciaria dubia (HBK) McVaugh] is a small fruit native to the Amazonian rain forest. Its anthocyanin profile has now been investigated for the first time. Fruits from two different regions of the S?o Paulo state, Brazil, were analyzed. The major anthocyanins were isolated by high-speed countercurrent chromatography. HPLC-PDA, HPLC-MS/MS, and 1H NMR were used to confirm the identity of the main anthocyanins of camu-camu. Cyanidin-3-glucoside was identified as the major pigment in the fruits from both regions, representing 89.5% in the fruits produced in Iguape and 88.0% in those from Mirandópolis, followed by the delphinidin-3-glucoside, ranging between 4.2 and 5.1%, respectively. Higher total anthocyanin contents were detected in the fruits from Iguape (54.0 +/- 25.9 mg/100 g) compared to those from Mirandópolis (30.3 +/- 6.8 mg/100 g), most likely because of the lower temperatures in the Iguape region.     

Molecular modeling of syringyl and p-hydroxyphenyl beta-O-4 dimers. Comparative study of the computed and experimental conformational properties of lignin beta-O-4 model compounds

As a new approach for the study of the ultrastructure of lignin, the conformational preferences of lignin beta-O-4 model compounds have been investigated by molecular modeling. The computed results have been compared with the experimental data (X-ray crystal structures and (3)J(H)(alpha)(H)(beta) NMR coupling constant values) reported in the literature. This comparison has led to an improved understanding of the influence of the structure, stereochemistry, and intramolecular H-bonding upon the conformational properties of the beta-O-4 dimers. A large number of low-energy conformations have been predicted for the structures. It has also appeared that the conformational features are predominantly governed by local steric interactions rather than by differences in the H-bonding interactions. The threo and erythro forms differ significantly in their conformational features, with a preferential extended overall shape for the threo form in which the bulky aromatic groups are distant from each other.     

Tritrichomonas foetus: a scanning electron microscopy study of erythrocyte adhesion associated with hemolytic activity

The in vitro hemolytic activity of Tritrichomonas foetus was investigated. The parasite was tested against human erythrocytes of groups A, B, AB, and O, and against erythrocytes of nine adult animals of different species (the rabbit, rat, chicken, cat, dog, swine, horse, bovine, and sheep). The results showed that T. foetus strains (ATCC KV1, K, PAL, 5022, RJ, 90) did not present any hemolytic activity against any human erythrocyte group nor against rabbit, rat, chicken, cat, dog and swine erythrocytes. T. foetus strains, however, lysed horse, bovine, and sheep erythrocytes. No hemolysin released by the parasites could be identified. Hemolysis did not occur with trichomonad culture supernatants, with sonicated extracts of T. foetus, nor with killed organisms. Scanning electron microscopy (SEM) showed that human erythrocytes did not adhere to the trophozoites, in contrast horse erythrocytes adhered to the surface of the parasites and were phagocytosed for up to 90 min. The parasites are able to exert their cytopathic effects through: (a) physical contact established between the two cell surfaces, (b) toxins released from parasites into the interaction media, or (c) the association of both mechanisms. Further studies are necessary to clarify the importance of the hemolytic activity in the biology of T. foetus.     

Voluntary intake and milk production in F1 Holstein × zebu cows in confinement

The objective of this study was to evaluate the nutrient intake and milk production in Holstein × zebu (F1) cows in feedlot. Eighteen F1 cows were used, divided into three treatments; six were Holstein × Gir (HGI), six were Holstein × Guzerat (HGU), and another six were Holstein × Nelore (HNE), which had recently calved, distributed into simple, random samples, under the same feeding conditions of corn silage and concentrate with 20% crude protein. The three-marker method was used (LIPE, titanium dioxide, and iADF) to estimate the individual intake and digestibility of the nutrients for the cows in group. The mathematical model used to establish the lactation curves was: Y = at(b)e(-ct) by Wood (Nature 216:164-165, 1967). The statistical analyses for the nutrient intake and digestibility, as well as parameters of metabolic efficiency, were performed using multiple linear regression (α = 5%). No effect (P > 0.05) of genetic group was observed for any of the variables studied. The intake and digestibility of the nutrients and the microbial nitrogen presented quadratic curves as a function of the lactation period (P < 0.05). The HGU cows exhibited an accumulated milk production of 4,946.81 kg at 305 days, whereas the HGI cows produced 4,821.78 kg. The HNE cows displayed inferior performance, with a production of 3,674.98 kg. It was concluded that, in confinement, F1 cows from different genetic groups do not exhibit different intake, digestibility, or metabolic efficiency. The HGU and HGI cows have greater cumulative production at 305 days.     

Update on the role of innate immune receptors during Brucella abortus infection

The innate immune system constitutes an efficient defense mechanism against invading microbial pathogens. Recent studies have revealed the intracellular signaling cascades involved in the TLR-initiated immune response to Brucella spp. infection. However, there is a piece of the puzzle missing that is the role of non-TLR receptors in innate immunity. The involvement of TLR receptors in brucellosis has been investigated by different research groups. It was demonstrated that TLR2 clearly does not play any role in controlling Brucella abortus infection in vivo, whereas TLR9 has been shown to be required for clearance of this bacterium in infected mice. The participation of adaptor molecules, such as MyD88 and TRIF has also been discussed. Recently, we and others have reported the critical role of MyD88- and not TRIF-mediated signaling in dendritic cell maturation and in vivo resistance during B. abortus infection. However, the relationship between specific Brucella molecules and non-TLR receptors and signal transduction pathways needs to be better understood. It is now clear that the interaction between TLRs and recently identified cytosolic innate immune sensors is crucial for mounting effective immune responses. Finally, this review discusses the mechanisms used by Brucella to escape detection by the host innate immune system.     

Mammary invasive micropapillary carcinoma in a male cat: Immunohistochemical description and clinical follow-up

We describe the case of a 14-year-old Domestic Short Hair male cat that presented with a single mammary tumour one centimetre in diameter, classified as invasive micropapillary carcinoma. Histology and immunohistochemistry revealed a high mitotic index, a KI-67 index of 10%, a lack of reactivity to myoepithelial markers, and a dense infiltration with T and B lymphocytes at the periphery of the tumour. Micrometastases were detected in the regional lymph node at the time of surgery. Overall survival time was 10 months with a disease-free interval of 7.5 months. Distant metastases in the sublumbar lymph nodes confirmed the aggressiveness of this tumour, which has recently been subtyped in female cats. This is the first case reported in male cats with a complete follow-up, highlighting the importance of prompt and aggressive treatment in the presence of mammary tumours in male cats.