A Combined Approach for the Assessment of Cell Viability and Cell Functionality of Human Fibrochondrocytes for Use in Tissue Engineering

Temporo-mandibular joint disc disorders are highly prevalent in adult populations. Autologous chondrocyte implantation is a well-established method for the treatment of several chondral defects. However, very few studies have been carried out using human fibrous chondrocytes from the temporo-mandibular joint (TMJ). One of the main drawbacks associated to chondrocyte cell culture is the possibility that chondrocyte cells kept in culture tend to de-differentiate and to lose cell viability under in in-vitro conditions. In this work, we have isolated human temporo-mandibular joint fibrochondrocytes (TMJF) from human disc and we have used a highly-sensitive technique to determine cell viability, cell proliferation and gene expression of nine consecutive cell passages to determine the most appropriate cell passage for use in tissue engineering and future clinical use. Our results revealed that the most potentially viable and functional cell passages were P5–P6, in which an adequate equilibrium between cell viability and the capability to synthesize all major extracellular matrix components exists. The combined action of pro-apoptotic (TRAF5, PHLDA1) and anti-apoptotic genes (SON, HTT, FAIM2) may explain the differential cell viability levels that we found in this study. These results suggest that TMJF should be used at P5–P6 for cell therapy protocols.     

Cellular types in the gill arch of the teleost Cyprinus carpio (Cyprinidae, Pisces). A histochemical and ultrastructural study

On adult specimens of the common carp (Cyprinus carpio), we have carried out a histochemical and ultrastructural study of the epithelia which form the gill arch. Secondary lamellae have two cellular types, granular and mucous, which produce neutral carbohydrates and proteins rich in tryptophane, and another mucous cell type which contains glycosaminoglycans. In gill rakers, three cellular types show different histochemical behaviour: 1) granular cells elaborate neutral mucosubstances; 2) a second cell type contains glycosaminoglycans,, and 3) a third cell type secretes neutral and acid carbohydrates. We discuss the possible role of these cells according to their secretion. We describe pillar and chloride cells in secondary lamellae, and chloride and neuroendocrine-like cells in gill rakers.     

Micropropagation of Rollinia mucosa (JACQ.) baill

Summary A protocol for in vitro propagation of Rollinia mucosa, an important medicinal plant, was developed. The presence of 500 mg l⁻¹ polyvinylpyrrolidone (PVP) during explant excision was important to avoid browning. Axillary buds, adventitious buds, and shoot cluster proliferation were achieved from epicotyl and hypocotyl explants from nursery-grown seedlings. The highest direct organogenesis percentage from hypocotyl explants was obtained upon culture of explants on Murashige and Skoog medium supplemented with 2.2 μM benzyladenine (BA) plus 2.32 μM kinetin. Epicotyl explants display highest regeneration frequency on a medium containing 8.8 μM BA and 0.54 μM naphthaleneacetic acid. Gibberellic acid was necessary for shoot elongation. Root induction was observed when shoots were pretreated with activated charcoal for 7 d in the dark before culture on Woody Plant Medium supplemented with 49.21 μM indolebutyric acid for 10 d. Root development was observed when 20 g l⁻¹ sucrose was used. Rooted plantlets were acclimatized and grown in the greenhouse.     

Utilization of food resources by small and medium‐sized mammals in the Monte Desert biome,Argentina

Theoretical models of species coexistence between desert mammals have generally been based on a combination of food and microhabitat selection by granivorous rodents. Although these models are applicable in various deserts of the world, they cannot explain resource use by mammals in Neotropical deserts. The present study examines diet composition in a mammal assemblage in the Monte desert, Argentina. The results show that two main strategies are used by these mammals: medium‐sized species (hystricognath rodents: Dolichotis patagonum, Lagostomus maximus, Microcavia australis and Galea musteloides; and an exotic lagomorph: Lepus europaeus) are herbivores, whereas small‐sized species (a marsupial: Thylamys pusillus; and sigmodontine rodents: Graomys griseoflavus, Akodon molinae, Calomys musculinus, Eligmodontia typus) are omnivorous. Small mammals also show a tendency towards granivory (C. musculinus), insectivory (A. molinae and T. pusillus) and folivory (G. griseoflavus).     

Using FTA® cards to store avian blood samples for genetic studies. Their application in sex determination

FTA® cards were used for long‐term storage of avian blood samples. Blood DNA was extracted by a simple method and used in PCR for sex identification of adult and nestling Great Grey Shrikes Lanius excubitor.     

Group versus population level demographics: An analysis of comparability using long term data on wild white‐faced capuchin monkeys (Cebus capucinus imitator)

Primates have long been used as indicator species for assessing overall ecosystem health. However, area‐wide census methods are time consuming, costly, and not always feasible under many field conditions. Therefore, it is important to establish whether monitoring a subset of a population accurately reflects demographic changes occurring in the population at large. Over the past 35 years, we have conducted 15 area‐wide censuses in Sector Santa Rosa, Costa Rica. These efforts have revealed important trends in population growth patterns of capuchin monkeys following the protection and subsequent regeneration of native forests. During this same period, we have also intensively studied a subset of the capuchin groups. Comparing these two datasets, we investigate whether the population structures of the closely monitored groups are reliable indicators of area‐wide demographic patterns. We compare the overall group size and the individual age/sex class compositions of study groups and nonstudy groups (i.e., those contacted during area‐wide censuses only). Our study groups contained more individuals overall with a larger proportion of infants, and there were indications that the proportion of adult and subadult males was lower. These differences can be ascribed either to sampling errors or real differences attributable to human presence and/or better habitat quality for the study groups. No other sex/age classes differed, and major demographic changes were simultaneously evident in both study and nonstudy groups. This study suggests that the Santa Rosa capuchin population is similarly impacted by large‐scale ecological patterns observable within our study groups.     

Metagenomic Analysis from the Interior of a Speleothem in Tjuv-Ante's Cave,Northern Sweden

Speleothems are secondary mineral deposits normally formed by water supersaturated with calcium carbonate percolating into underground caves, and are often associated with low-nutrient and mostly non-phototrophic conditions. Tjuv-Ante’s cave is a shallow-depth cave formed by the action of waves, with granite and dolerite as major components, and opal-A and calcite as part of the speleothems, making it a rare kind of cave. We generated two DNA shotgun sequencing metagenomic datasets from the interior of a speleothem from Tjuv-Ante’s cave representing areas of old and relatively recent speleothem formation. We used these datasets to perform i) an evaluation of the use of these speleothems as past biodiversity archives, ii) functional and taxonomic profiling of the speleothem’s different formation periods, and iii) taxonomic comparison of the metagenomic results to previous microscopic analyses from a nearby speleothem of the same cave. Our analyses confirm the abundance of Actinobacteria and fungi as previously reported by microscopic analyses on this cave, however we also discovered a larger biodiversity. Interestingly, we identified photosynthetic genes, as well as genes related to iron and sulphur metabolism, suggesting the presence of chemoautotrophs. Furthermore, we identified taxa and functions related to biomineralization. However, we could not confidently establish the use of this type of speleothems as biological paleoarchives due to the potential leaching from the outside of the cave and the DNA damage that we propose has been caused by the fungal chemical etching.     

Development of hepatic acetyl coenzyme A carboxylase in hormone-treated chicks.

It has been suggested that the carbohydrate-rich diet of chicks after hatching is responsible for the emergence of hepatic enzymes involved in lipogenesis; the injection of glucose to newly hatched chicks gives rise to an appreciable elvation on the activities of acetyl coenzyme A carboxylase and fatty acid synthetase. The present study shows that during the first hours after hatching, there is a natural elevation of glycemia which parallels the increase in acetyl coenzyme A carboxylase activity. However, the administration of hormones which alter the blood glucose levels considerably (insulin, tolbutamide, glucagon and hydrocortisone) did not influence the enzyme activity. The administration of thyroxine, estradiol and cyclic AMP, was also without effect. These results do not support the theory that the increased amount of blood glucose is the natural effector of the induction acetyl coenzyme A carboxylase. They also show that different lipogenic enzymes are not regulated via the same 'operon' since thyroxine or glucagon which alter the level of some enzymes on this pathway did not modify that of the acetyl coenzyme A carboxylase.     

Demonstrating the Potential for Dynamic Auditory Stimulation to Contribute to Motion Sickness

Auditory cues can create the illusion of self-motion (vection) in the absence of visual or physical stimulation. The present study aimed to determine whether auditory cues alone can also elicit motion sickness and how auditory cues contribute to motion sickness when added to visual motion stimuli. Twenty participants were seated in front of a curved projection display and were exposed to a virtual scene that constantly rotated around the participant''s vertical axis. The virtual scene contained either visual-only, auditory-only, or a combination of corresponding visual and auditory cues. All participants performed all three conditions in a counterbalanced order. Participants tilted their heads alternately towards the right or left shoulder in all conditions during stimulus exposure in order to create pseudo-Coriolis effects and to maximize the likelihood for motion sickness. Measurements of motion sickness (onset, severity), vection (latency, strength, duration), and postural steadiness (center of pressure) were recorded. Results showed that adding auditory cues to the visual stimuli did not, on average, affect motion sickness and postural steadiness, but it did reduce vection onset times and increased vection strength compared to pure visual or pure auditory stimulation. Eighteen of the 20 participants reported at least slight motion sickness in the two conditions including visual stimuli. More interestingly, six participants also reported slight motion sickness during pure auditory stimulation and two of the six participants stopped the pure auditory test session due to motion sickness. The present study is the first to demonstrate that motion sickness may be caused by pure auditory stimulation, which we refer to as “auditorily induced motion sickness”.