Altered Cortical Palmitoylation Induces Widespread Molecular Disturbances in Parkinson’s Disease

The relationship between Parkinson’s disease (PD), the second-most common neurodegenerative disease after Alzheimer’s disease, and palmitoylation, a post-translational lipid modification, is not well understood. In this study, to better understand the role of protein palmitoylation in PD and the pathways altered in this disease, we analyzed the differential palmitoyl proteome (palmitome) in the cerebral cortex of PD patients compared to controls (n = 4 per group). Data-mining of the cortical palmitome from PD patients and controls allowed us to: (i) detect a set of 150 proteins with altered palmitoylation in PD subjects in comparison with controls; (ii) describe the biological pathways and targets predicted to be altered by these palmitoylation changes; and (iii) depict the overlap between the differential palmitome identified in our study with protein interactomes of the PD-linked proteins α-synuclein, LRRK2, DJ-1, PINK1, GBA and UCHL1. In summary, we partially characterized the altered palmitome in the cortex of PD patients, which is predicted to impact cytoskeleton, mitochondrial and fibrinogen functions, as well as cell survival. Our study suggests that protein palmitoylation could have a role in the pathophysiology of PD, and that comprehensive palmitoyl-proteomics offers a powerful approach for elucidating novel cellular pathways modulated in this neurodegenerative disease.     

Functional characteristics of pyruvate transport in Phycomyces blakesleeanus

A saturable and accumulative transport system for pyruvate has been detected in Phycomyces blakesleeanus NRRL 1555(-) mycelium. It was strongly inhibited by alpha-cyano-4-hydroxycinnamate. l-Lactate and acetate were competitive inhibitors of pyruvate transport. The initial pyruvate uptake velocity and accumulation ratio was dependent on the external pH. The Vmax of transport greatly decreased with increasing pH, whereas the affinity of the carrier for pyruvate was not affected. The pyruvate transport system mediated its homologous exchange, which was essentially pH independent, and efflux, which increased with increasing external pH. The uptake of pyruvate was energy dependent and was strongly inhibited by inhibitors of oxidative phosphorylation and of the formation of proton gradients. Glucose counteracted the inhibitory effect of the pyruvate transport produced by inhibitors of mitochondrial ATP synthesis. Our results are consistent with a pyruvate/proton cotransport in P. blakesleeanus probably driven by an electrochemical gradient of H+ generated by a plasma membrane H+-ATPase.     

Metataxonomic and immunological analysis of milk from ewes with or without a history of mastitis

Mastitis is a highly prevalent condition that has a great impact on milk production and animal welfare, and often requires substantial management efforts. For this reason, it is generally considered an important threat to the dairy industry. Many microbial, host, and environmental factors can protect against, predispose to, or influence the development of mastitis. The objective of this work was to characterize the milk microbiota of Manchega ewes, and to compare samples from animals with and without a history of mastitis. We analyzed milk samples from 36 ewes belonging to 2 different farms (18 ewes from each farm) using culture-dependent and culture-independent techniques. We also analyzed several immune compounds to investigate associations of mastitis with 3 main variables: farm; history of mastitis or no mastitis; and parity number. Both culture-dependent and culture-independent techniques showed that ewe milk harbored a site-specific complex microbiota and microbiome. Staphylococcus epidermidis was the main species driving the difference between farm A (where it was the dominant species) and B (where it was not). In contrast, samples from farm B were characterized by the presence of a wide spectrum of other coagulase-negative staphylococci. Some of these species have already been associated with subclinical intramammary infections in ruminants. Of the 10 immune compounds assayed in this study, 3 were related to a history of mastitis [IL-8, IFN-γ, and IFN-gamma-induced protein 10 (IP-10)]. Increases in IL-8 concentrations in milk seemed to be a feature of subclinical mastitis in sheep, and in this study, this immune factor was detected only in samples from ewes with some episodes of mastitis and from the group with the highest somatic cell count. We also observed a positive correlation between the samples with the highest somatic cell count and IFN-γ and IP-10 levels. Our results suggest that these 3 compounds could be used as biomarkers for the negative selection of mastitis-prone animals, particularly when somatic cell count is very high.     

The applied pretreatment (blanching,ascorbic acid) at the manufacture process affects the quality of nectarine purée processed by hydrostatic high pressure

A nectarine purée was manufactured with different pretreatments (thermal blanching or ascorbic acid – AA – addition), and then, the purée was processed by high‐pressure treatment to evaluate the effect of the initial manufacture conditions in the stability of the processed purées. A thermal treatment was also carried out to compare the effect with the high‐pressure processing (HPP). All applied processes were effective to ensure the microbiological safety of the purées. However, the pretreatment (thermal blanching or AA addition) applied during the manufacture affected the final quality of the processed purées. Initially, the AA addition had a protective effect on colour degradation during the manufacture of the purées; however, when these purées were treated by HPP showed less colour stability during storage, lower bioactive compounds content, and antioxidant activity. In contrast, purées with an initial thermal blanching maintained better the quality after HPP and during storage.     

Functional characterization of human and fungal MAP kinases in Saccharomyces cerevisiae

A functional comparison of three human SAPKs with fungal Hog1p was undertaken, using Saccharomyces cerevisiae as a heterologous expression system. We characterized the role of mammalian MAP kinases in sensitivity to both osmotic and oxidative stress of a S. cerevisiae hog1 mutant. Western blot analyses indicated that S. cerevisiae can only phosphorylate mammalian MAP kinases in response to osmotic stress but not to oxidative stress, while morphogenetic defects characteristic of hog1 mutants under hyperosmotic stress are only suppressed by fungal and not mammalian Hog1p. Our data demonstrate the functional conservation of MAPKs although they also evidence differential aspects among the three human SAPKs and the fungal MAPKs.