Isolation, characterization and biological evaluation of jellyfish collagen for use in biomedical applications

Fibrillar collagens are the more abundant extracellular proteins. They form a metazoan-specific family, and are highly conserved from sponge to human. Their structural and physiological properties have been successfully used in the food, cosmetic, and pharmaceutical industries. On the other hand, the increase of jellyfish has led us to consider this marine animal as a natural product for food and medicine. Here, we have tested different Mediterranean jellyfish species in order to investigate the economic potential of their collagens. We have studied different methods of collagen purification (tissues and experimental procedures). The best collagen yield was obtained using Rhizostoma pulmo oral arms and the pepsin extraction method (2-10 mg collagen/g of wet tissue). Although a significant yield was obtained with Cotylorhiza tuberculata (0.45 mg/g), R. pulmo was used for further experiments, this jellyfish being considered as harmless to humans and being an abundant source of material. Then, we compared the biological properties of R. pulmo collagen with mammalian fibrillar collagens in cell cytotoxicity assays and cell adhesion. There was no statistical difference in cytotoxicity (p > 0.05) between R. pulmo collagen and rat type I collagen. However, since heparin inhibits cell adhesion to jellyfish-native collagen by 55%, the main difference is that heparan sulfate proteoglycans could be preferentially involved in fibroblast and osteoblast adhesion to jellyfish collagens. Our data confirm the broad harmlessness of jellyfish collagens, and their biological effect on human cells that are similar to that of mammalian type I collagen. Given the bioavailability of jellyfish collagen and its biological properties, this marine material is thus a good candidate for replacing bovine or human collagens in selected biomedical applications.     

Polyoxygenated Steroids from the Octocoral Leptogorgia punicea and in Vitro Evaluation of Their Cytotoxic Activity

Five new polyoxygenated marine steroids—punicinols A–E (1–5)—were isolated from the gorgonian Leptogorgia punicea and characterized by spectroscopic methods (IR, MS, ¹H, ¹³C and 2-D NMR). The five compounds induced in vitro cytotoxic effects against lung cancer A549 cells, while punicinols A and B were the most active, with IC₅₀ values of 9.7 μM and 9.6 μM, respectively. The synergistic effects of these compounds with paclitaxel, as well as their effects on cell cycle distribution and their performance in the clonogenic assay, were also evaluated. Both compounds demonstrated significant synergistic effects with paclitaxel.     

Ecotoxicological risk assessment of a quarry filling with seaport sediments using laboratory freshwater aquatic microcosms

Purpose

In France, contaminated seaport sediments cannot be discharged into the sea according to recent regulation. Hence, they must be managed on land. Among the solutions identified, that of filling terrestrial quarries located in the littoral zone is one of the most promising. However, this requires developing a methodology for ecological risk assessment; which was the aim of the SEDIGEST research program. In the present study, we report the results of toxicological impacts of three sediments on aquatic ecosystems close to a quarry. These results were used to formulate a risk assessment methodology.

Materials and methods

The ecotoxicological approach was based on laboratory microcosm assays applied to leachates obtained from the sediments. The microcosms contained synthetic water and sediment and five pelagic (micro-algae, duckweeds and daphnids) and benthic (chironomids and amphipods) freshwater species. The biota were exposed for 3 weeks to a range of leachate concentrations; metals were monitored in the water column and the effects (i.e. mortality, growth inhibition and midge emergence) were measured.

Results and discussion

The results showed an absence of acute toxicity at concentrations of <10% (v/v) but sublethal effects for concentrations between 1 and 10%. Given the complex nature of the leachates, it was difficult to determine the factors of toxicity. Metals, especially Cu, might have been responsible for some of the effects on the amphipods.

Conclusions

Future quarries filled with seaport sediments might emit pollutants to aquatic ecosystems in their vicinity, and risk assessors should ensure that predicted environmental concentrations of leachates are below the maximum leachate concentration acceptable for the whole ecosystem; for example, by multiplying the concentration safe for the microcosm organisms by a factor of 10.     

Silicon alleviates the toxicity of cadmium and zinc for maize (Zea mays L.) grown on a contaminated soil

Although silicon (Si) is not an essential element, it presents a close relationship with the alleviation of heavy‐metal toxicity to plants. This work was carried out to evaluate the effects of Si application to soil on the amelioration of metal stress to maize grown on a contaminated soil amended with Si (0, 50, 100, 150, and 200 mg kg^–1) as calcium silicate (CaSiO₃). Additionally, the cadmium (Cd) and zinc (Zn) bioavailability as well as their distribution into soil fractions was also studied. The results showed that adding Si to a Cd‐ and Zn‐contaminated soil effectively diminished the metal stress and resulted in biomass increase in comparison to metal‐contaminated soil not treated with Si. This relied on Cd and Zn immobilization in soil rather than on the increase of soil pH driven by calcium silicate application. Silicon altered the Cd and Zn distribution in soil fractions, decreasing the most bioavailable pools and increasing the allocation of metals into more stable fractions such as organic matter and crystalline iron oxides.     

Composition of beer by 1H NMR spectroscopy: effects of brewing site and date of production

A principal component analysis (PCA) of 1H NMR spectra of beers differing in production site (A, B, C) and date is described, to obtain information about composition variability. First, lactic and pyruvic acids contents were found to vary significantly between production sites, good reproducibility between dates being found for site A but not for sites B and C beers. Second, site B beers were clearly distinguished by the predominance of linear dextrins, while A and C beers were richer in branched dextrins. Carbohydrate reproducibility between dates is poorer for site C with dextrin branching degree varying significantly. Finally, all production sites were successfully distinguished by their contents in adenosine/inosine, uridine, tyrosine/tyrosol, and 2-phenylethanol, reproducibility between dates being again poorer for site C. Interpretation of the above compositional differences is discussed in terms of the biochemistry taking place during brewing, and possible applications of the method in brewing process control are envisaged.     

Hematological and splenic Doppler ultrasonographic changes in dogs sedated with acepromazine or xylazine

Objective

To evaluate the onset and duration of hematological changes and the use of Doppler ultrasound (spleen) in dogs sedated with acepromazine or xylazine.

Variation in pestivirus growth in testicle primary cell culture is more dependent on the individual cell donor than cattle breed

The causes of bovine respiratory disease complex (BRDC) are multifactorial and include infection with both viral and bacterial pathogens. Host factors are also involved as different breeds of cattle appear to have different susceptibilities to BRDC. Infection with bovine pestiviruses, including bovine viral diarrhea virus 1 (BVDV1), BVDV2 and ‘HoBi’-like viruses, is linked to the development of BRDC. The aim of the present study was to compare the growth of different bovine pestiviruses in primary testicle cell cultures obtained from taurine, indicine and mixed taurine and indicine cattle breeds. Primary cells strains, derived from testicular tissue, were generated from three animals from each breed. Bovine pestivirus strains used were from BVDV-1a, BVDV-1b, BVDV-2a and ‘HoBi’-like virus. Growth was compared by determining virus titers after one passage in primary cells. All tests were run in triplicate. Virus titers were determined by endpoint dilution and RT-qPCR. Statistical analysis was performed using one way analysis of variance (ANOVA) followed by the Tukey’s Multiple Comparison Test (P?0.05). Significant differences in virus growth did not correlate with cattle breed. However, significant differences were observed between cells derived from different individuals regardless of breed. Variation in the replication of virus in primary cell strains may reflect a genetic predisposition that favors virus replication.     

Cover Image,Volume 73, Issue 1

The cover image, by Oscar Fernando Santos‐Amaya et al., is based on the Research Article Fitness costs and stability of Cry1Fa resistance in Brazilian populations of Spodoptera frugiperda, DOI: 10.1002/ps.4312 . Photo Credit: Dr. Oscar Santos‐Amaya.

     

Efficiency of methods for genetic progress estimation in common bean breeding using database information

The final field trials to evaluate elite lines developed by the Embrapa national common bean breeding program generated a phenotypic database composed by agronomic traits of 84 elite lines and nine cultivars over a 16-year period (1993–2008) and 450 environments in all Brazilian growing areas. The main goal of this study was to use this database as a model to compare the consistency of the results obtained from indirect methods for genetic progress estimation for grain yield in common bean breeding, using the direct method as a reference. Three indirect methods for genetic progress estimation were evaluated: (1) linear regression with unadjusted averages, (2) linear regression with averages adjusted by the mixed models, and (3) linear regression with averages adjusted by a fixed effects model with the error exception. The genetic progress estimated by the direct method was 31.3 kg ha^?1 per year (1.34%**). This value was considered as the reference estimate, since it was calculated using the grain yield data from final field trials with all common bean lines evaluated under the same environmental conditions. The estimate obtained using the regression with unadjusted averages of the three best lines by cycle was 25.66 kg ha^?1 per year (1.26%*), similar to the result obtained by the direct method. Considering both methods using fixed and mixed models, the genetic gain estimates were statistically null (0.42% and 0.45%, respectively). Therefore, the regression method with unadjusted means was more informative than the other indirect methods.