Selective Cytotoxicity of 1,3,4-Thiadiazolium Mesoionic Derivatives on Hepatocarcinoma Cells (HepG2)

In this work, we evaluated the cytotoxicity of mesoionic 4-phenyl-5-(2-Y, 4-X or 4-X-cinnamoyl)-1,3,4-thiadiazolium-2-phenylamine chloride derivatives (MI-J: X=OH, Y=H; MI-D: X=NO₂, Y=H; MI-4F: X=F, Y=H; MI-2,4diF: X=Y=F) on human hepatocellular carcinoma (HepG2), and non-tumor cells (rat hepatocytes) for comparison. MI-J, M-4F and MI-2,4diF reduced HepG2 viability by ~ 50% at 25 μM after 24-h treatment, whereas MI-D required a 50 μM concentration, as shown by 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyltetrazolium bromide assays. The cytotoxicity was confirmed with lactate dehydrogenase assay, of which activity was increased by 55, 24 and 16% for MI-J, MI-4F and MI-2,4diF respectively (at 25 μM after 24 h). To identify the death pathway related to cytotoxicity, the HepG2 cells treated by mesoionic compounds were labeled with both annexin V and PI, and analyzed by flow cytometry. All compounds increased the number of doubly-stained cells at 25 μM after 24 h: by 76% for MI-J, 25% for MI-4F and MI-2,4diF, and 11% for MI-D. It was also verified that increased DNA fragmentation occurred upon MI-J, MI-4F and MI-2,4diF treatments (by 12%, 9% and 8%, respectively, at 25 μM after 24 h). These compounds were only weakly, or not at all, transported by the main multidrug transporters, P-glycoprotein, ABCG2 and MRP1, and were able to slightly inhibit their drug-transport activity. It may be concluded that 1,3,4-thiadiazolium compounds, especially the hydroxy derivative MI-J, constitute promising candidates for future investigations on in-vivo treatment of hepatocellular carcinoma.     

Evolutionary trends in the chromosome numbers of swarm-founding social wasps

Chromosome number and morphology are relevant aspects of genomic organization of eukaryotes and are considered key components to comprehend evolutionary mechanisms and karyotypic differentiation. Social wasps belonging to the tribe Epiponini show complex social characteristics that make these insects interesting models for genetic and evolutionary studies. However, there is a paucity of genetic information in social wasps as a whole. Aiming to investigate the process of chromosomal evolution of the social Epiponini wasps, chromosomes of ten species of this group were analyzed using Giemsa and fluorochrome staining techniques and fluorescence in situ hybridization in two species. In this study a high variation in the chromosome number and morphology was found and the previous range of chromosome number of Epiponini was broadened from n = 8–32 to n = 5–33. We also suggest that chromosomal segments with high GC content must have had a key role in the karyotype diversification of these wasps. Moreover, based on a phylogenetic background we find evidence of a main role of chromosomal fusion in the occurrence of gradual decrease of chromosome number in Epiponini during its chromosomal evolutionary history.     

Saccharomyces cerevisiae strain UFMG 905 protects against bacterial translocation, preserves gut barrier integrity and stimulates the immune system in a murine intestinal obstruction model

Probiotic is a preparation containing microorganisms that confers beneficial effect to the host. This work assessed whether oral treatment with viable or heat-killed yeast Saccharomyces cerevisiae strain UFMG 905 prevents bacterial translocation (BT), intestinal barrier integrity, and stimulates the immunity, in a murine intestinal obstruction (IO) model. Four groups of mice were used: mice undergoing only laparotomy (CTL), undergoing intestinal obstruction (IO) and undergoing intestinal obstruction after previous treatment with viable or heat-killed yeast. BT, determined as uptake of ^99mTc-E. coli in blood, mesenteric lymph nodes, liver, spleen and lungs, was significantly higher in IO group than in CTL group. Treatments with both yeasts reduced BT in blood and all organs investigated. The treatment with both yeasts also reduced intestinal permeability as determined by blood uptake of ^99mTc-DTPA. Immunological data demonstrated that both treatments were able to significantly increase IL-10 levels, but only viable yeast had the same effect on sIgA levels. Intestinal lesions were more severe in IO group when compared to CTL and yeasts groups. Concluding, both viable and heat-killed cells of yeast prevent BT, probably by immunomodulation and by maintaining gut barrier integrity. Only the stimulation of IgA production seems to depend on the yeast viability.     

Urinary Excretion of Zinc and Metabolic Control of Patients with Diabetes Type 2

The objective of this study was to assess urinary excretion of zinc and evaluation parameters of metabolic control in type 2 diabetic patients. Thirty-one type 2 diabetic patients, of both genders, with 5.8 ± 5.6 years average time of the disease, age range 20–60 years, were selected. Evaluation of the nutritional status was performed using anthropometric measurements. To evaluate food consumption, the 3-day alimentary log method was used, and its analysis was performed using a software. Determination of urinary zinc was by atomic absorption spectrophotometry. From the obtained results, it was concluded that 51.6% of the patients were overweight. The mean of found waist circumference was 100.4 and 92.2 cm for men and women, respectively. Blood glucose and glycated hemoglobin values were higher than reference values, and plasma albumin concentration was adequate. The median of found urinary zinc excretion was 474.9 μg/24 h, within normal standards (300–600 μg/day). Regarding diet composition, calorie and protein concentration were above recommendation, while mean zinc concentration was adequate. This data allow the conclusion that the evaluated patients presented adequate urinary zinc excretion in comparison with reference values.     

Gas exchange and water relations of three <Emphasis Type="Italic">Vitis vinifera</Emphasis> L. cultivars growing under Mediterranean climate

Optical characteristics, contents of photosynthetic pigments, total soluble sugars, and starch, rates of gas exchange, chlorophyll (Chl) a fluorescence, and leaf water relations were analysed in three Vitis vinifera L. cultivars, Tinto Cão (TC), Touriga Nacional (TN), and Tinta Roriz (TR), grown in Mediterranean climate. Chl content was significantly lower in TC than in TN and TR leaves, while the Chl a/b ratio was higher. TR had the lowest net photosynthetic rate, stomatal conductance, and contents of soluble sugars and starch than TN and TC. In spite of low Chl content, TC showed the lowest photon absorbance and the highest photochemical efficiency of photosystem 2. TC had the lowest predawn and midday leaf water potential. The capability for osmotic adjustment was similar among cultivars and the calculated modulus of elasticity was higher in TC leaves. The typical lighter green leaves of TC seemed to be an adaptive strategy to high irradiance and air temperature associated to water stress.     

Western scrub-jays anticipate future needs independently of their current motivational state

Planning for the future has been considered to be a uniquely human trait [1-3]. However, recent studies challenge this hypothesis by showing that food-caching Western scrub-jays (Aphelocoma californica) can relate their previous experience as thieves to the possibility of future cache theft by another bird [4], are sensitive to the state of their caches at recovery ([5] and S. De Kort, S.P.C.C., D. Alexis, A.D., and N.S.C., unpublished data), and can plan for tomorrow's breakfast [6]. Although these results suggest that scrub-jays are capable of future planning, the degree to which these birds act independently of their current motivational state is a matter of contention. The Bischof-K?hler hypothesis [1] holds that nonhuman animals cannot anticipate and act toward the satisfaction of a future need not currently experienced or cued by their present motivational state. Using specific satiety to control for the jays' current and future motivational states, here we specifically test this hypothesis by dissociating current and future motivational states. We report that Western scrub-jays anticipate the recovery of their caches, as well as their own future needs, by acting independently of their current motivational state and immediate needs. The fact that the birds act in favor of a future need as opposed to the current one challenges the hypothesis that this ability is unique to humans.     

Liver response of the Antarctic fish (<Emphasis Type="Italic">Notothenia coriiceps</Emphasis> Richardson, 1844) to hepatotrophic factors

The Notothenia coriiceps liver is commonly infected with parasites, reducing the hepatic mass and inducing the regeneration. In order to better understand the effect of nutrient influx on hepatic regeneration at 0°C, a usual mammal hepatotrophic factor (HF) solution was injected into ten fish (HF group), while ten fish were injected with saline solution (control), twice a day, for 15 days. The liver and carcass weight were measured, and samples were obtained for histological studies. The HF group presented a higher liver/carcass weight (62.5%) than control group. This increase in liver mass was due mainly to hepatocytes hypertrophy, including nuclear size increase and cytoplasmic inclusions of glycogen. Hyperplasia is also observed, although to a lesser extent. The hepatic reaction to HF in Antarctic fish was here demonstrated for the first time, helping to understand the liver response to seasonal nutrient.     

Seed bank dynamics of two obligate seeders, <Emphasis Type="Italic">Cistus monspeliensis</Emphasis> and <Emphasis Type="Italic">Rosmarinus officinalis</Emphasis>, in relation to time since fire

Many species in Mediterranean-type ecosystems regenerate after fire by seed germination from soil seed banks. Seed bank dynamics of two of those obligate seeders, Cistus monspeliensis and Rosmarinus officinalis, were investigated in relation to stand age since fire in southwestern Portugal. Soil seed density, annual seed input, annual seed losses through germination and seed persistence were compared between species at stands differing in age since fire (5, 10 and 35 years). Soil seed density and seed input increased over the first decade after fire and were lowest at 35-year-old stands for C. monspeliensis. In R. officinalis, few seeds were produced and found in the soil at early stages, and maximum seed input and soil seed density were attained at 35-year-old stands. Soil seed density was mostly driven by seed production in both species, which is largely dependent on plant traits and population dynamics related to fire. Overall, stand age since fire had a negligible effect on seed germination, seed persistence and viability. Ten to 39% of buried seeds were not recovered after 1 year, and viability of seeds recovered was 97–100% for C. monspeliensis and only 0–3% for R. officinalis. Variation in plant traits within the seeder syndrome was evidenced by this study. R. officinalis evidenced lower seed persistence, lower proportion of viable seed produced and lower density of viable soil seed than C. monspeliensis at any stage after fire. R. officinalis is expected to depend largely on previous year seed production for population replacement after fire.     

In situ synthesis of magnetite nanoparticles in carrageenan gels

Magnetite nanoparticles have been successfully synthesized in the presence of carrageenan polysaccharides using an in situ coprecipitation method. Iron coordination to the sulfate groups of the polysaccharide was confirmed by FTIR. The polysaccharide type (kappa, iota, or lambda) and concentration have been varied and their effects on particle morphology and chemical stability of the resultant nanocomposite investigated. The presence of carrageenan induces the formation of smaller particles, compared to those formed in the absence of polymer, and their average size depends on the nature and concentration of the polysaccharide used. The chemical stability of magnetite nanoparticles toward oxidation was also seen to depend on biopolymer type with magnetite formed in iota-carrageenan showing the highest chemical stability. A general tendency toward lower stability is observed as the polysaccharide concentration is increased. It is suggested that magnetite chemical stability in the carrageenan composites is determined by a fine balance between particle size and gel strength, the latter determining oxygen diffusion rates through the medium.