Structural and Biological Characterization of Two Crotamine Isoforms IV-2 and IV-3 Isolated from the <Emphasis Type="Italic">Crotalus durissus cumanensis</Emphasis> Venom

In this work, we isolated the two new crotamine isoforms from the Crotalus durissus cumanensis rattlesnake venom and its “in vitro” neurotoxic, myotoxic and lethality (DL₅₀) intracerebroventricular (i.c.v.) effects were characterized. These proteins were named IV-2 and IV-3 and were purified by combination of two chromatographic steps on molecular exclusion chromatography on Superdex 75 and reverse phase HPLC (μ-Bondapack C18). The molecular mass of the crotamine isoforms was 4905.96 Da for isoform IV-2 and 4956.97 Da for IV-3 and, as determined by mass spectrometry, and both contained six Cys residues. Enzymatic hydrolysis followed by de novo sequencing by tandem mass spectrometry was used to determine the primary structure of both isoforms. The positions of five sequenced tryptic peptides, including the N-terminal of the isoform IV-2 and four from isoform IV-3 were deduced by comparison with a homologous protein from the crotamine family. The isoforms IV-2 and IV-3 had a sequence of amino acids of 42 amino acid residues IV-2: YKRCHIKGGH CFPKEKLICI PPSSDIGKMD CPWKRKCCKK RS and pI value 9.54 and IV-3: YKQCHKKGGH CFPKEVLICI PPSSDFGKMD CRWKRKCCKK RS with a pI value of 9.54. This protein showed high molecular amino acid sequence identity with other crotamine-like proteins from Crotalus durissus terrificus. These new crotamine isoforms induced potent blockade of neuromuscular transmission in young chicken biventer cervicis preparation and potent myotoxic effect. In mice, both isoforms induced myonecrosis, upon intramuscular or subcutaneous injections. These activities were modulated by the presence of positively charged amino acid residues. The LD₅₀ of isoform IV-2 was 0.07 mg/kg and isoform IV-3 was 0.06 mg/kg the animal weight, by i.c.v. route.     

Floristic composition and similarity of 15 hectares in Central Amazon, Brazil

The Amazon region is one of the most diverse areas in the world. Research on high tropical forest diversity brings up relevant contributions to understand the mechanisms that result and support such diversity. In the present study we describe the species composition and diversity of 15 one-ha plots in the Amazonian terra firme dense forest in Brazil, and compare the floristic similarity of these plots with other nine one-ha plots. The 15 plots studied were randomly selected from permanent plots at the Embrapa Experimental site, Amazonas State in 2005. The diversity was analysed by using species richness and Shannon's index, and by applying the Sorensen's index for similarity and unweighted pair-group average (UPGMA) as clustering method. Mantel test was performed to study whether the differences in species composition between sites could be explained by the geographic distance between them. Overall, we identified 8 771 individuals, 264 species and 51 plant families. Most of the species were concentrated in few families and few had large number of individuals. Families presenting the highest species richness were Fabaceae (Faboideae: 22spp., Mimosoideae: 22spp.), Sapotaceae: 22spp., Lecythidaceae: 15 and Lauraceae: 13. Burseraceae had the largest number of individuals with 11.8% of the total. The ten most abundant species were: Protium hebetatum (1 037 individuals), Eschweilera coriacea (471), Licania oblongifolia (310), Pouteria minima (293), Ocotea cernua (258), Scleronema micranthum (197), Eschweilera collina (176), Licania apelata (172), Naucleopsis caloneura (170) and Psidium araca (152), which represented 36.5% of all individuals. Approximately 49% of species had up to ten individuals and 13% appeared only once in all sampled plots, showing a large occurrence of rare species. Our study area is on a forest presenting a high tree species diversity with Shannon's diversity index of 4.49. The dendrogram showed two groups of plots with low similarity between them (less than 0.25), and the closer the plots were one to another, more similar in species composition (Mantel R = 0.3627, p < 0.01). The 15 plots in our study area share more than 50% of their species composition and represent the group of plots that have the shortest distance between each other. Overall, our results highlight the high local and regional heterogeneity of environments in terra firme forests, and the high occurrence of rare species, which should be considered in management and conservation programs in the Amazon rainforest, in order to maintain its structure on the long run.     

Direct restorative treatment of anterior weared teeth after re‐establishment of occlusal vertical dimension: a case report

doi: 10.1111/j.1741‐2358.2012.00630.x Direct restorative treatment of anterior weared teeth after re‐establishment of occlusal vertical dimension: a case report Objective: The loss of posterior teeth resulting in a lack of stability may lead to excessive mandibular load on some points of occlusion of the remaining teeth, causing wear. The rehabilitation of these cases must be carefully planned and the treatment requires a period of adaptation with a new vertical dimension. Background: This case report describes the treatment of a patient with a loss of almost all posterior inferior teeth and a consequential loss of occlusal vertical dimension and accentuated wear of anterior elements. Materials and methods: Provisional removable partial dentures were manufactured and the patient used them for a period of adaptation prior to restoration of the anterior teeth. After the patient’s adaptation with the new occlusal dimension, anterior restorations were made using a mycrohibrid resin composite with different color aspects for an optimal stratification. Results: The restorations achieved an aesthetic configuration with ideal function as the final result and the occlusion showed a satisfactory stability for a provisional removable partial dentures. Conclusion: This article presents an anterior direct rehabilitation approach not only as an alternative option, but also as viable and less expensive treatment option with an optimally aesthetic and functional result.     

The Role of Lipotoxicity in Smoke Cardiomyopathy

Background/Aims

Experimental and clinical studies have shown the direct toxic effects of cigarette smoke (CS) on the myocardium, independent of vascular effects. However, the underlying mechanisms are not well known.

Methods

Wistar rats were allocated to control (C) and cigarette smoke (CS) groups. CS rats were exposed to cigarette smoke for 2 months.

Results

After that morphometric, functional and biochemical parameters were measured. The echocardiographic study showed enlargement of the left atria, increase in the left ventricular systolic volume and reduced systolic function. Within the cardiac metabolism, exposure to CS decreased beta hydroxy acyl coenzyme A dehydrogenases and citrate synthases and increased lactate dehydrogenases. Peroxisome proliferator-activated receptor alpha (PPARα) and peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α) were expressed similarly in both groups. CS increased serum lipids and myocardial triacylglycerols (TGs). These data suggest that impairment in fatty acid oxidation and the accumulation of cardiac lipids characterize lipotoxicity. CS group exhibited increased oxidative stress and decreased antioxidant defense. Finally, the myocyte cross-sectional area and active Caspase 3 were increased in the CS group.

Conclusion

The cardiac remodeling that was observed in the CS exposure model may be explained by abnormalities in energy metabolism, including lipotoxicity and oxidative stress.     

Diet of the gracile mouse opossum (Gracilinanus microtarsus) (Didelphimorphia: Didelphidae) in a Brazilian cerrado: patterns of food consumption and intrapopulation variation

The diet of the gracile mouse opossum Gracilinanus microtarsus was studied in a cerrado remnant in south-eastern Brazil through the analysis of faeces sampled from adult individuals. Patterns of food resource consumption were assessed using the statistics of per cent occurrence. Intrapopulation variation in the number of food items detected in faeces as a function of relevant factors was inferred using generalized linear models. The latter statistical formalism also allowed variation in the number of food items detected in faeces to be interpreted in terms of rate ratios among levels of significant factors. Insects, spiders, snails and fruits were detected in the faeces of G. microtarsus , with insects, particularly termites, beetles and ants, being the most frequently detected food resource. These food resources were consumed in proportion to their relative abundance in the cerrado, suggesting that G . microtarsus is an opportunistic forager feeding primarily on insects. The generalized linear model identified sex, season and food resource as significant factors affecting the number of food items detected in faeces, as well as interactions between sex and season and season and food resource. Rate ratios calculated between sexes within seasons showed that the number of food items detected in the faeces of males was larger than that detected in the faeces of females in the warm-wet and cool-dry seasons. Rate ratios calculated between seasons within sexes showed different trends in the number of food items detected in faeces for each significant food resource. It is suggested that differences in the number of food items detected in faeces between sexes within season and between seasons within sexes are related to energetic requirements associated with reproduction.     

Antifungal and physicochemical properties of Ocimum essential oil loaded in poly(lactic acid) nanofibers

Poly(lactic acid) (PLA) nanofibres containing different proportions of the essential oils from Ocimum basilicum L. and Ocimum gratissimum L. were prepared by solution blow spinning method. The essential oils were extracted by hydrodistillation and characterized by gas chromatography. MEV, contact angle, DSC and FTIR were used to characterize the nanofibres. The effect of bioative nanofibres on the growth of the fungus and on the production of ochratoxin A were evaluated using the fumigation test. Linalool, 1·8-cineole and camphor were the principal components of the essential oil from O. basilicum, and eugenol was the principal constituent in the oil from O. gratissimum. An increase in the average diameter of the nanofibres was observed with the addition of the essential oils. The essential oils acted as a plasticizer, resulting in a reduction in the crystallinity of the PLA. The encapsulation of essential oils in PLA nanofibres was verified by FTIR. An effective antifungal and antimicotoxygenic activity against Aspergillus ochraceus and Aspergillus westerdjikiae was observed for the bioative nanofibres. These results confirm the potential of PLA nanofibres containing the essential oils for the control of toxigenic fungi that cause the deterioration of food and are harmful to human health.     

Glycerol binding at the narrow channel of photosystem II stabilizes the low-spin S2 state of the oxygen-evolving complex

Photosynthesis Research - The oxygen-evolving complex (OEC) of photosystem II (PSII) cycles through redox intermediate states Si (i?=?0–4) during the photochemical oxidation of...     

Plant sex and phenological stage affect interactions with rhizosphere nematode communities

Background: In dioecious plants, females and males associate differently with mycorrhizal fungi, but interactions with other rhizosphere organisms are less well studied.

Aims: We investigated the effect of plant sex on rhizosphere nematode communities associated with Corema album, a dioecious shrub occurring in coastal habitats.

Methods: Rhizosphere samples were collected from males and females in three populations (150 plants), during fruiting and flowering. Nematode communities were characterised and compared between plant sexes through statistical analyses of the abundance of trophic groups, plant parasitic nematode (PPN) genera and ecological indices.

Results: Free-living nematodes showed no statistically significant differences owing to plant sex. Conversely, PPN community composition was significantly different between plant sexes during fruiting but not flowering, suggesting that physiological requirements over the annual phenological cycle of the plant influence ecological interactions with the rhizosphere.

Of the 13 PPN genera identified, the ectoparasitic Criconema and Hemicriconemellawere more abundant in the rhizosphere of males during fruiting, whereas the endoparasitic Meloidogyne associated more frequently with females, suggesting that plant host suitability is related to PPN feeding strategy.

Conclusions: It appears that interactions of individuals of different sexes of C. album with the rhizosphere nematode community vary with phenological stage, especially for PPN.     

Diagrammatic scale for severity evaluation of the Eremanthus erythropappus–Puccinia velata pathosystem

Candeia (Eremanthus erythropappus (DC.) MacLeish), a native forest species from South America, has garnered commercial interest due to its production of essential oil that contains alpha-bisabolol. This compound is widely used in the pharmaceutical and cosmetics industry, with approximately 80% of Brazilian production being exported. Since candeia rust (Puccinia velata) has only been reported in Brazil, little is known about its epidemiology and control. There is no methodology to quantify rust severity in candeia, justifying the elaboration and validation of a diagrammatic scale containing eight levels of disease severity based on leaf area coverage (0.25%, 0.5%, 1%, 2%, 4%, 8%, 16% and 32%). In a natural sampling of disease in the field, 95% of the leaves showed severity below 16%, with the remaining 5% showing severities between 16% and 32% of leaf area. Validation of the proposed diagrammatic scale was performed by assessing the results from 10 inexperienced evaluators, performing evaluations of three leaves with different severity levels. The evaluations were performed at 7-day intervals; in the first instance, severity values were assigned without the diagrammatic scale, and for the second attempt, the scale proposed in this study was used. The accuracy and precision of the severity estimates produced by each evaluator compared to the real severity was analysed by linear regression and by Lin's statistics. The reproducibility of the estimates was evaluated by analysing the coefficient of determination of linear regressions by pairs of evaluators. The scale provided adequate levels of accuracy, precision, repeatability and reproducibility, indicating the proposed scale was a suitable method for quantifying the severity of candeia rust.