In vitro replication of spontaneous fractures of the proximal human femur

Spontaneous fractures (i.e. caused by sudden loading and muscle contraction, not by trauma) represent a significant percentage of proximal femur fractures. They are particularly relevant as may occur in elderly (osteoporotic) subjects, but also in relation to epiphyseal prostheses. Despite its clinical and legal relevance, this type of fracture has seldom been investigated. Studies concerning spontaneous fractures are based on a variety of loading scenarios. There is no evidence, nor consensus on the most relevant loading scenario. The aim of this work was to develop and validate an experimental method to replicate spontaneous fractures in vitro based on clinically relevant loading. Primary goals were: (i) repeatability and reproducibility, (ii) clinical relevance. A validated numerical model was used to identify the most critical loading scenario that can lead to head-neck fractures, and to determine if it is necessary to include muscle forces when the head-neck region is under investigation. The numerical model indicated that the most relevant loading scenario is when the resultant joint force is applied to the head at 8 degrees from the diaphysis. Furthermore, it was found that it is not essential to include the muscles when investigating head-neck fractures. The experimental setup was consequently designed. The procedure included high-speed filming of the fracture event. Clinically relevant fracture modes were obtained on 10 cadaveric femurs. Failure load should be reported as a fraction of donor's body-weight to reduce variability. The proposed method can be used to investigate the reason and mechanism of failure of natural and operated proximal femurs.     

Sensory ecology of ostariophysan alarm substances

Chemical communication of predation risk has evolved multiple times in fish species, with conspecific alarm substance (CAS) being the most well understood mechanism. CAS is released after epithelial damage, usually when prey fish are captured by a predator and elicits neurobehavioural adjustments in conspecifics which increase the probability of avoiding predation. As such, CAS is a partial predator stimulus, eliciting risk assessment-like and avoidance behaviours and disrupting the predation sequence. The present paper reviews the distribution and putative composition of CAS in fish and presents a model for the neural processing of these structures by the olfactory and the brain aversive systems. Applications of CAS in the behavioural neurosciences and neuropharmacology are also presented, exploiting the potential of model fish [e.g., zebrafish Danio rerio, guppies Poecilia reticulata, minnows Phoxinus phoxinus) in neurobehavioural research.     

Bacterial endocarditis and increased cardiac troponin I levels in a brown howler monkey (Alouatta guariba clamitans) with an interventricular septal defect

A brown howler monkey (Alouatta guariba clamitans) was presented with lethargy, hyporexia, cough and heart murmur. The complementary tests and necropsy revealed pleuropneumonia, bacterial endocarditis and interventricular septal defect. To the best of our knowledge, this is the first report of increased cardiac troponin I levels in this species.     

Comparative Analysis of the Antibacterial Activity and HPLC Phytochemical Screening of the Brazilian Red Propolis and the Resin of Dalbergia ecastaphyllum

The aim of this study was to investigate the antibacterial activity of red propolis and resin and their association with standard antibiotics to evaluate possible differences of activity. We also submitted red propolis and the resin to a HPLC analysis to confirm the botanical origin. The extracts were tested against P. aeruginosa and S. aureus alone and in association with gentamicin and imipenem. The HPLC analysis identified seven compounds with six of them present in both substances. The lowest MIC values obtained in this study were observed against S. aureus. In general, MIC values showed to be lower for red propolis against all species tested in comparison to resin. Despite the synergistic behavior to be similar for both substances, we observed that inhibitory concentrations of drugs were lower when associated with red propolis in comparison to resin.     

In Vitro Identification of Histatin 5 Salivary Complexes

With recent progress in the analysis of the salivary proteome, the number of salivary proteins identified has increased dramatically. However, the physiological functions of many of the newly discovered proteins remain unclear. Closely related to the study of a protein’s function is the identification of its interaction partners. Although in saliva some proteins may act primarily as single monomeric units, a significant percentage of all salivary proteins, if not the majority, appear to act in complexes with partners to execute their diverse functions. Coimmunoprecipitation (Co-IP) and pull-down assays were used to identify the heterotypic complexes between histatin 5, a potent natural antifungal protein, and other salivary proteins in saliva. Classical protein–protein interaction methods in combination with high-throughput mass spectrometric techniques were carried out. Co-IP using protein G magnetic Sepharose TM beads suspension was able to capture salivary complexes formed between histatin 5 and its salivary protein partners. Pull-down assay was used to confirm histatin 5 protein partners. A total of 52 different proteins were identified to interact with histatin 5. The present study used proteomic approaches in conjunction with classical biochemical methods to investigate protein–protein interaction in human saliva. Our study demonstrated that when histatin 5 is complexed with salivary amylase, one of the 52 proteins identified as a histatin 5 partner, the antifungal activity of histatin 5 is reduced. We expected that our proteomic approach could serve as a basis for future studies on the mechanism and structural-characterization of those salivary protein interactions to understand their clinical significance.     

Sodium Transport in Turtle Erythrocytes : Apparent stimulation of exchange diffusion by anaerobiosis

Studies were performed on Na and K transport by red blood cells of the freshwater turtle under anaerobic and aerobic conditions. Although it had previously been assumed that cation transport in turtle red blood cells was dependent on respiration, the present data show greater Na efflux rates in N₂ than in O₂. However, ouabain inhibited Na transport by the same amount quantitatively in O₂ and N₂ gas phases. Thus there was no difference in ouabain-sensitive or "pump" Na transport rates. Na influx rates were higher in nitrogen than in air and potassium influx rates were not significantly different under aerobic and anaerobic conditions. Moreover in the absence of sodium in the bathing medium no difference between air and nitrogen could be discovered. Finally with ethacrynic acid plus ouabain there was an additional decrease in Na efflux but there was a persisting difference between air and nitrogen. These studies do not rule out the existence of a ouabain-insensitive ethacrynic acid-inhibitable flux; however, they suggest that at least part of the activation of Na efflux observed in N₂ was due to increased exchange diffusion.     

Morpho–anatomical and ultrastructural analysis of extrafloral nectaries in Inga edulis (Vell.) Mart. (Leguminosae)

The presence of extrafloral nectaries (EFNs) between leaflets is an usual feature in Inga edulis (Vell.) Mart. (Leguminosae). Extrafloral nectaries are secretory structures involved in production of nectar and which serve in the protection of plants against herbivores through association with ants. This study aimed to characterize the EFNs of I. edulis at different developmental stages and describe their morphology, histochemistry and ultrastructure. Leaf fragments, containing secretory structures, were processed according to standard methods for light, scanning and transmission electron microscopy. The EFNs were classified into three stages based on morphology: pre‐secretory, secretory and post‐secretory. The pre‐secretory stage occurs in young leaves, whereas secretory and post‐secretory stages occur in developed and senescent leaves, respectively. The EFNs possess a concave surface and a central cleft in which nectar is accumulated and which was not observed in pre‐secretory EFNs. Histochemical tests identified the presence of sugars, proteins, phenolic compounds, mucilage and lipids at all developmental stages of the EFNs. Calcium crystals were identified in all tissues and stages of the EFNs. The secretory cells of the EFNs exhibit a granular cytoplasm, small vacuoles, prominent nuclei, smooth endoplasmic reticulum and mitochondria. Post‐secretory stage EFNs exhibited intense cytoplasmic degradation and the presence of microorganisms. The performance of EFNs of I. edulis appear to follow the leaf development.     

Phenolic composition and antiparasitic activity of plants from the Brazilian Northeast “Cerrado”

This work describes the antiparasitic and cytotoxic activities of three plant species from the Cerrado biome, Northeastern Brazil. Significant antiparasitic inhibition was observed against Trypanosoma cruzi (63.86%), Leishmania brasiliensis (92.20%) and Leishmania infantum (95.23%) when using ethanol extract from leaves of Guazuma ulmifolia Lam. (Malvaceae), at a concentration of 500 μg/mL. However, low levels of inhibition were observed when assessing leishmanicidal and trypanocidal (Clone CL-B5) activities of crude ethanol extracts from leaves and bast tissue of Luehea paniculata (Malvaceae) and leaves and bark of Prockia crucis (Salicaceae) at a concentration of 500 μg/mL. The extracts revealed the presence of phenolic acids such as gallic acid, chlorogenic acid, caffeic acid and rosmarinic acid, as well as flavonoids such as rutin, luteolin, apigenin and quercetin – the latter detected only in G. ulmifolia. G. ulmifolia extract displayed higher leishmanicidal activity probably due to the presence of quercetin, a potent known leishmanicidal compound. A cytotoxicity test indicated values over 50% at the highest concentration (1000 μg/mL) for all natural products, which were considered cytotoxic. This points out the need for further tests to enable future in vivo trials, including antineoplastic activity on human tumor cells.     

The occurrence of the red-handed howler monkey (Alouatta belzebul) in amazonian savannas is related to forest patch area and density of flooded area palms

All Neotropical primates are arboreal and thus depend on forests for their survival. Arboreality puts many Neotropical primates at risk of extinction due to the high rates of deforestation in the tropics. We assessed the influence of vegetation structure and forest patch attributes on the occurrence of the threatened red-handed howler monkey (Alouatta belzebul) in an Amazonian savanna. Using a sample of 38 forest patches in a region of approximately 2000 km² in the state of Amapá, northern Brazil, we used logistic regression to find the best predictors of the occurrence of A. belzebul. We assessed patch area, patch isolation, the proportion of seasonally flooded forest in the patch, the density of flooded area palms, forest height, canopy cover, and diameter at breast height of trees. Patch area and palm density were the best predictors of the occurrence of A. belzebul in forest patches, both having a positive effect on the probability of occurrence. Our results indicate that areas of flooded forest in forest patches may be keystone habitats for A. belzebul living in Amazonian savannas. The observed effect of palm density on A. belzebul suggests that this variable is useful for planning conservation actions, including the selection of areas for protection and management strategies for areas inhabited by this primate.