Preservation at -195 degrees C of trypsinized organs for primary cultures

Trypsinized cells from rabbit kidney can be kept at -195, 8 degrees C indefinitely. When necessary, propagation in monolayer cultures is readily obtained as primary cultures, provided that some conditions be respected: 1) only glass culture flask must be used. Falcon plastic vessels do not allow cell attachment in this type of primary culture 2) the lag period is particularly long: it taken about 20 days before the beginning of cell multiplication 3) fetal calf serum is toxic for the cells in the 2nd and 3rd change of the culture medium, during the lag period.     

Native drivers of fish life history traits are lost during the invasion process

Rapid adaptation to global change can counter vulnerability of species to population declines and extinction. Theoretically, under such circumstances both genetic variation and phenotypic plasticity can maintain population fitness, but empirical support for this is currently limited. Here, we aim to characterize the role of environmental and genetic diversity, and their prior evolutionary history (via haplogroup profiles) in shaping patterns of life history traits during biological invasion. Data were derived from both genetic and life history traits including a morphological analysis of 29 native and invasive populations of topmouth gudgeon Pseudorasbora parva coupled with climatic variables from each location. General additive models were constructed to explain distribution of somatic growth rate (SGR) data across native and invasive ranges, with model selection performed using Akaike's information criteria. Genetic and environmental drivers that structured the life history of populations in their native range were less influential in their invasive populations. For some vertebrates at least, fitness‐related trait shifts do not seem to be dependent on the level of genetic diversity or haplogroup makeup of the initial introduced propagule, nor of the availability of local environmental conditions being similar to those experienced in their native range. As long as local conditions are not beyond the species physiological threshold, its local establishment and invasive potential are likely to be determined by local drivers, such as density‐dependent effects linked to resource availability or to local biotic resistance.     

Quantitative RT-PCR assay for VEGF mRNA in human tumors of the kidney

Angiogenesis is the formation of new capillaries from pre-existing vessels, and recent evidence has demonstrated that tumor growth is controlled mainly by angiogenesis. Vascular endothelial growth factor (VEGF) is an endothelium-specific growth factor which is strongly angiogenic in vitro and in vivo. We have developed a quantitative RT-PCR assay for the measurement of VEGF mRNA expression using a real-time procedure based on the use of fluorogenic probes and the ABI PRISM 7700 Sequence Detector System. The assay performance of this method in terms of practicability and reliability is reported with results that seem promising for its widespread use in the clinical laboratory. The method has been applied to the measurement of mRNA of VEGF in human renal cell carcinomas (RCC). Preliminary results show a significantly higher VEGF mRNA expression (ratio values between 181 and 2222) in tumor specimens compared to non-adjacent, non-tumoral tissue of the same subjects.     

Fermentation of wheat: effects of backslopping different proportions of pre-fermented wheat on the microbial and chemical composition

The objective of the study was to examine effect of backslop on the chemical and microbiological characteristics of fermented wheat (FW). Coarsely ground wheat was mixed with water (1:3 wt/wt) and inoculated with 6 log cfu ml(-1) each of an overnight culture of Lactobacillus plantarum and Pediococcus pentosaceus. Four fermentation treatments were conducted in 45 1, closed, PVC containers over 48 hours. Three treatments investigated the benefits of the addition of previously fermented wheat (backslopping, BSL) at different proportions (0.20, 0.33 or 0.42 kg) to freshly prepared wheat. The control treatment contained no addition of BSL. Elimination of coliforms from the FW within 48 h was only achieved through backslopping; where coliform bacteria counts decreased from approximately 6.5 log10 cfu ml(-1) to less than 3 log10 cfu ml(-1). There was no apparent advantage in increasing the backslop proportion above 0.20. However, the exclusion of coliform bacteria required the pH to remain below 4.0 for at a minimum of 24 h. The results of these studies indicate that fermentation of wheat has the potential to reduce the risk of feed-borne colibacillosis and provides a practical alternative to producers that cannot ferment multiple diets or have limited fermentation capacity.     

Discovery of a series of imidazopyrazine small molecule inhibitors of the kinase MAPKAPK5, that show activity using in vitro and in vivo models of rheumatoid arthritis

MAPKAPK5 has been proposed to play a role in regulation of matrix metalloprotease expression and so to be a potential target for intervention in rheumatoid arthritis. We present here the identification of a series of compounds against this target which are effective in both biochemical and cell assays. The expansion of the series is described, along with early SAR and pharmacokinetics for some representative compounds.     

Morphological traits determine the winner of “symmetric” fights in hermit crabs

This study aimed at investigating in the hermit crab Pagurus longicarpus whether some morphological traits and their slight variations might determine the winner of a contest in symmetric pairs, i.e. pairs composed of individuals matched by their overall size. In the pre-experimental phase, 400 crabs were individually kept in isolation for a week to eliminate the effects of their previous social experiences and were allowed to enter adequate shells to equalize their motivation to fight. Then, we formed 200 pairs matched for shell and body size and observed their agonistic behaviour for 15 min. Alphas (and betas) were deemed as those individuals that won more than half of the recorded fights. Finally, crabs were sacrificed and sexed, and 6 and 8 measures were taken of their shells and bodies, respectively. Raw data were converted to compositional data and subjected to an isometric log-ratio transformation prior to statistical analysis. Alphas were found to occupy heavier and wider shells than betas, both characteristics that, together with the correlated high level of encrustation, make the apparent dimension of the crab larger and thus favour that individual during agonistic encounters. As expected, hermit crabs were sexually dimorphic in their chelae, but sexes differed also in the length of the dactylus of their third walking leg and in their body weight. Alphas were heavier than betas and had longer propodi and wider dactyli on the third walking legs than betas. The latter character might be advantageous during shell fights when the walking legs of the attacker are used to hold the defender's shell in the spasmodic shaking and their wider dactyli might assure a firmer seizing of it. Indeed, a correct execution of this pattern allows for the eviction of the defender. Our results suggest that more attention towards variations in morphological traits other than body size might improve our understanding of the factors that affect individual Resource Holding Power and fight dynamics.     

Animal xenodiversity in Italian inland waters: distribution, modes of arrival, and pathways

The paper provides a list of the non-indigenous animal species occurring today in Italian inland waters. Xenodiversity was found to amount to 112 species (64 invertebrates and 48 vertebrates), which contribute for about 2% to the inland-water fauna in Italy. Northern and central regions are most affected, and Asia, North America, and the rest of Europe are the main donor continents. The large majority of non-indigenous species entered Italy as a direct or indirect effect of human intervention. A difference between invertebrates and vertebrates was found for their mode of arrival (unintentional for invertebrates and intentional for vertebrates). Accidental transport, in association with both fish (for aquaculture or stock enhancement) and crops, has been the main vector of invertebrate introductions, whereas vertebrates were mostly released for stocking purposes. Overall stock enhancement (47.92%) and culture (37.5%) prevailed over the other pathways. Seventeen and 7 species of our list are included among the 100 worst invasive species of Europe (DAISIE) and of the world (IUCN), respectively. For some (but not all) non-indigenous species recorded in Italy the multilevel impact exerted on the recipient communities and ecosystems is known, even if rarely quantified, but knowledge on their chronic impact is still missing. Additional research is needed to provide criteria for prioritizing intervention against well established invaders and identify which new potential invader should be targeted as “unwanted”.     

Shell recruitment in the Mediterranean hermit crab Clibanarius erythropus

Gastropod shells are vital for the majority of hermit crab species, being essential for their survival, growth, protection, and reproduction. Given their importance, shells are acquired and transferred between crabs through several modalities. We conducted observations and experiments at the Asinara Island (Sardinia, Italy) to investigate the efficacy of the different behavioral tactics adopted by the hermit crab Clibanarius erythropus to acquire shells, such as: (1) locomotion and activity at different tidal phases; (2) attendance at shell-supplying sites (simulated predation sites with five different odors: live and dead gastropods, live and dead crabs, predator); and (3) interactions with conspecifics in aggregations on simulated gastropod predation sites. In each tidal phase, locomotion was slow (0.7 cm min^− 1) and, as a consequence, the probability of encountering empty shells and conspecifics was low. Simulated gastropod predation sites quickly attracted a larger number of hermit crabs than the other sites tested. Aggregations seemed to function as shell exchange markets, as previously suggested for other species: the first attendant took the experimental shell and a chain of shell exchanges among conspecifics followed. Our results show that, in C. erythropus, aggregation is the most efficient tactic for the acquisition of new shells, whereas in other species, such as Pagurus longicarpus, it is associated with exploitation ability due to the intense locomotion. The interspecific plasticity in hermit crabs' behavior is confirmed.     

Bioimpedancemetry for the assessment of periodontal tissue inflammation: a numerical feasibility study

In dentistry possible inflammatory episodes of oral cavity can be very frequent (periodontitis, mucositis, peri-implantitis) and they can have serious consequences. Indeed, peri-implantitis is still the principal cause of implant failure. Impedance values of biological tissues are related to the physiological/pathological state of the tissue itself. In fact, an inflamed site exhibits an impedance value lower than that of the corresponding healthy tissue. Based on these observations, the aim of this work is to determine if impedancemetric measurements are able to provide information about the inflammatory state of tissues. A numerical 3D model has been realized to simulate the measurement conditions present in the event of inflammation around a dental implant. The aim is to understand if it is possible to determine the presence of an inflamed tissue and to locate its site, so that the treatment could be specifically focused in that specific area. A simplified geometry reproducing the implant has been realized in order to validate the numerical model by means of experimental measurements. The obtained results are satisfactorily accurate, so the model can be considered reliable. Therefore, multiple simulations have been run on the original model to carry out a parametric study in terms of different conductivity values, different volumes of inflamed tissues and different measurement frequencies. The advantages and limits of such a method have been shown to properly define the main constraints for the system design.