Insertion of Pex5p into the peroxisomal membrane is cargo protein-dependent

It is now generally accepted that Pex5p, the receptor for most peroxisomal matrix proteins, cycles between the cytosol and the peroxisomal compartment. According to current models of peroxisomal biogenesis, this intracellular trafficking of Pex5p is coupled to the transport of newly synthesized peroxisomal proteins into the organelle matrix. However, direct evidence supporting this hypothesis was never provided. Here, using an in vitro peroxisomal import system, we show that insertion of Pex5p into the peroxisomal membrane requires the presence of cargo proteins. Strikingly the peroxisomal docking/translocation machinery is also able to catalyze the membrane insertion of a Pex5p truncated molecule lacking any known cargo-binding domain. These results suggest that the cytosol/peroxisomal cycle in which Pex5p is involved is directly or indirectly regulated by Pex5p itself and not by the peroxisomal docking/translocation machinery.     

Osteoclast cell-surface changes during the egg-laying cycle in Japanese quail

The medullary bone serves as a source of labile calcium mobilized during calcification of the egg shell in birds. Quantitative histological methods demonstrate that the numbers of medullary bone osteoclasts and nuclei per osteoclast remain unchanged during the egg cycle in the Japanese quail (Coturnix). Therefore, cyclic changes in bone resorption cannot be explained by modulations of osteoclasts from and into other bone cells, a mechanism previously suggested for certain species of birds. Rather, dramatic changes in osteoclast cell-surface features occur during the egg cycle, which might account for cyclic variations in resorptive activity. During egg shell calcification, osteoclasts with ruffled borders are closely apposed to bone surfaces; the cytoplasm is rich in vacuoles that contain mineral crystals and seem to derive from the ruffled border. At the completion of egg shell calcification, the ruffled borders and vacuoles move away from the bone surface, although the osteoclast remains attached to the bone along the filamentous or "clear" zone. Associated with the disappearance of the ruffled borders is the appearance of extensive interdigitated cell processes along the peripheral surface of the osteoclast away from the bone. These unusual structures, which may serve as a reservoir of membrane, largely disappear when ruffled borders and associated structures reappear. Therefore, in these hens, the osteoclasts modulate their cell surface rather than their population during the egg cycle.     

Carotenoid and lipid production by the autotrophic microalga Chlorella protothecoides under nutritional, salinity, and luminosity stress conditions

Today microalgae represent a viable alternative source for high-value products. The specie Chlorella protothecoides (Cp), heterotrophically grown, has been widely studied and provides a high amount of lutein and fatty acids (FA) and has a good profile for biodiesel production. This work studies carotenoid and FA production by autotrophic grown Cp. Cp was grown until the medium’s nitrogen was depleted, then diluted in NaCl solution, resulting in nutritional, luminosity, and salinity stresses. Different NaCl concentrations were tested (10, 20, 30 g/L) at two different dilutions. After dilution, a color shifting from green to orange-red was noticed, showing carotenoid production. The best production of both carotenoids and FA was attained with a 20 g/L NaCl solution. The total carotenoid content was 0.8 % w/w (canthaxanthin (23.3 %), echinenone (14.7 %), free astaxanthin (7.1 %), and lutein/zeaxanthin (4.1 %)). Furthermore, the total lipid content reached 43.4 % w/w, with a FA composition of C18:1 (33.64 %), C16:0 (23.30 %), C18:2 (11.53 %), and less than 12 % of C18:3, which is needed to fulfill the biodiesel quality specifications (EN 14214).     

Long-term pattern of population dynamics in the field vole from central Europe: cyclic pattern with amplitude dampening

The subject of population cycles is regarded as controversial due to a number of unsettled questions such as whether or not cyclic patterns are governed by the same processes at high and low latitudes in Europe. Recent evidence suggests that the dynamics at high and low latitudes share the common temporal pattern of vole dynamics referred to as collapsing population cycles. Despite concurrent interest, the key contention around the causal mechanisms that drive population cycles remains a hot topic in ecology. The aims of this study are to supplement information on the seasonal population dynamics of the field vole Microtus agrestis in the Czech Republic by analysing 25 years of time series data. By applying robust estimation procedures, we estimated several parameters to describe population dynamics, such as population variability, amplitude dampening, cycle period, order of the dynamics and the structure of density dependence. The parameters indicate that field vole dynamics in central Europe are highly variable, cyclic dynamics of order two, with peaks in abundance occurring regularly at intervals of 4–5 years. In addition to exhibiting population cycles, the field vole populations show a pattern of dampened amplitude as observed elsewhere in Europe, including northern latitudes. By analysing temporal trends in seasonal abundances, population growth rates and environmental temperatures, we did not obtain evidence to support the hypothesis that amplitude dampening results from the negative effect of increasingly mild winters on winter population growth rates.     

Nematicidal action of chitinases produced by the fungus Monacrosporium thaumasium under laboratorial conditions

Nematophagous fungi produce chitinases that may be important in the process of infection of eggs and larvae of nematodes. This study aimed to produce, purify, characterise and test the nematicidal action of extracellular chitinases produced by Monacrosporium thaumasium on Panagrellus redivivus. Mycelia from M. thaumasium were used to inoculate a solid medium for chitinase production. The enzymes were purified using a specific technique of adsorption for chitinases. The chitinase activity was determined at different pHs and temperatures. NF34 produced two distinct chitinases (27 and 30 kDa). After 72 hours, these enzymes provided a significant reduction (80%; p < 0.01) of the number of P. redivivus larvae, compared to control. It was shown that isolate NF34 produced chitinases with nematicidal activity. Thus, other experimental designs on geohelminths or even arthropods that transmit diseases may become a new aspect of the field of study of biological control using predatory nematophagous fungi.     

Understanding the structural and dynamic consequences of DNA epigenetic modifications: Computational insights into cytosine methylation and hydroxymethylation

We report a series of molecular dynamics (MD) simulations of up to a microsecond combined simulation time designed to probe epigenetically modified DNA sequences. More specifically, by monitoring the effects of methylation and hydroxymethylation of cytosine in different DNA sequences, we show, for the first time, that DNA epigenetic modifications change the molecule''s dynamical landscape, increasing the propensity of DNA toward different values of twist and/or roll/tilt angles (in relation to the unmodified DNA) at the modification sites. Moreover, both the extent and position of different modifications have significant effects on the amount of structural variation observed. We propose that these conformational differences, which are dependent on the sequence environment, can provide specificity for protein binding.     

Trophic relationship between the Patinga hybrid (Piaractus mesopotamicus x Piaractus brachypomus) and Echinorhynchus gomesi Machado Filho, 1948 in fish farms

A total of 122 Patinga specimens were collected from fish farms (P1, P2 and P3), and only those from fish farm P1 were shown to be infected with Echinorhynchus gomesi. In addition, fish in this study were shown to have diets that consisted of 21 different food items, and Notodiaptomus sp. (Copepoda: Calanoida) was identified as a potential intermediate host for E. gomesi.     

T(H)17-based vaccine design for prevention of Streptococcus pneumoniae colonization

Streptococcus pneumoniae is a leading cause of mortality in young children. While successful conjugate polysaccharide vaccines exist, a less expensive serotype-independent protein-based pneumococcal vaccine offers a major advancement for preventing life-threatening pneumococcal infections, particularly in developing nations. IL-17A-secreting CD4+ T cells (T(H)17) mediate resistance to mucosal colonization by multiple pathogens including S. pneumoniae. Screening an expression library containing >96% of predicted pneumococcal proteins, we identified antigens recognized by T(H)17 cells from mice immune to pneumococcal colonization. The identified antigens also elicited IL-17A secretion from colonized mouse splenocytes and human PBMCs suggesting that similar responses are primed during natural exposure. Immunization of two mouse strains with identified antigens provided protection from pneumococcal colonization that was significantly diminished in animals treated with blocking CD4 or IL-17A antibodies. This work demonstrates the potential of proteomic screening approaches to identify specific antigens for the design of subunit vaccines against mucosal pathogens via harnessing T(H)17-mediated immunity.