Comparative proteome analysis reveals that blood and sugar meals induce differential protein expression in Aedes aegypti female heads.

Aedes aegypti females ingest sugar or blood to obtain the nutrients needed to maintain cellular homeostasis. During human blood ingestion, female mosquitoes may transmit different viruses such as dengue, yellow fever and, more recently, zika and chikungunya. Here, we report changes in protein expression in the heads of adult female Ae. aegypti mosquitoes in response to the ingestion of blood or sugar. Proteins extracted from the heads of Ae. aegypti fed exclusively on blood (BF) or sugar (SF) were trypsin hydrolyzed (off-gel) and analyzed by the reverse-phase nano-liquid chromatography coupled with hybrid mass spectrometry. A total of 1139 proteins were identified in female heads, representing 7.4% of the predicted proteins in Ae. aegypti genome (total = 15 419 active genes). Gene ontology annotation and categories showed that, in this insect, the head was rich in proteins involved in the metabolic process, proton transport, organelle, macromolecular complex, structural molecule activity, antioxidant activity, and catalytic activity. Our report is the first indicating that many of the annotated genes are translated into functional proteins in heads of adult female Ae. aegypti. Interestingly, we identified 8.7 times more exclusively expressed proteins involved in signal transduction, replication-transcription-translation (5.5 x), and transport (2.9 x) activity in BF than in SF groups. This paper discusses the protein profile of Ae. aegypti female heads and its implications for blood ingestion and carbohydrate intake.

Oligomerization and Membrane-binding Properties of Covalent Adducts Formed by the Interaction of α-Synuclein with the Toxic Dopamine Metabolite 3,4-Dihydroxyphenylacetaldehyde (DOPAL).

Oxidative deamination of dopamine produces the highly toxic aldehyde 3,4-dihydroxyphenylacetaldehyde (DOPAL), enhanced production of which is found in post-mortem brains of Parkinson disease patients. When injected into the substantia nigra of rat brains, DOPAL causes the loss of dopaminergic neurons accompanied by the accumulation of potentially toxic oligomers of the presynaptic protein α-synuclein (aS), potentially explaining the synergistic toxicity described for dopamine metabolism and aS aggregation. In this work, we demonstrate that DOPAL interacts with aS via formation of Schiff-base and Michael-addition adducts with Lys residues, in addition to causing oxidation of Met residues to Met-sulfoxide. DOPAL modification leads to the formation of small aS oligomers that may be cross-linked by DOPAL. Both monomeric and oligomeric DOPAL adducts potently inhibit the formation of mature amyloid fibrils by unmodified aS. The binding of aS to either lipid vesicles or detergent micelles, which results in a gain of α-helix structure in its N-terminal lipid-binding domain, protects the protein against DOPAL adduct formation and, consequently, inhibits DOPAL-induced aS oligomerization. Functionally, aS-DOPAL monomer exhibits a reduced affinity for small unilamellar vesicles with lipid composition similar to synaptic vesicles, in addition to diminished membrane-induced α-helical content in comparison with the unmodified protein. These results suggest that DOPAL could compromise the functionality of aS, even in the absence of protein oligomerization, by affecting the interaction of aS with lipid membranes and hence its role in the regulation of synaptic vesicle traffic in neurons.

Proteome analysis of plastids from developing seeds of Jatropha curcas L.

In this study, we performed a proteomic analysis of plastids isolated from the endosperm of developing Jatropha curcas seeds that were in the initial stage of deposition of protein and lipid reserves. Proteins extracted from the plastids were digested with trypsin, and the peptides were applied to an EASY-nano LC system coupled inline to an ESI-LTQ-Orbitrap Velos mass spectrometer, and this led to the identification of 1103 proteins representing 804 protein groups, of which 923 proteins were considered as true identifications, and this considerably expands the repertoire of J. curcas proteins identified so far. Of the identified proteins, only five are encoded in the plastid genome, and none of them are involved in photosynthesis, evidentiating the nonphotosynthetic nature of the isolated plastids. Homologues for 824 out of 923 identified proteins were present in PPDB, SUBA, or PlProt databases while homologues for 13 proteins were not found in any of the three plastid proteins databases but were marked as plastidial by at least one of the three prediction programs used. Functional classification showed that proteins belonging to amino acids metabolism comprise the main functional class, followed by carbohydrate, energy, and lipid metabolisms. The small and large subunits of Rubisco were identified, and their presence in the plastids is considered to be an adaptive feature counterbalancing for the loss of one-third of the carbon as CO2 as a result of the conversion of carbohydrate to oil through glycolysis. While several enzymes involved in the biosynthesis of several precursors of diterpenoids were identified, we were unable to identify any terpene synthase/cyclase, which suggests that the plastids isolated from the endosperm of developing seeds do not synthesize phorbol esters. In conclusion, our study provides insights into the major biosynthetic pathways and certain unique features of the plastids from the endosperm of developing seeds at the whole proteome level.

Are gastric cancer resection margin proteomic profiles more similar to those from controls or tumors?

A strategy for treating cancer is to surgically remove the tumor together with a portion of apparently healthy tissue surrounding it, the so-called "resection margin", to minimize recurrence. Here, we investigate whether the proteomic profiles from biopsies of gastric cancer resection margins are indeed more similar to those from healthy tissue than from cancer biopsies. To this end, we analyzed biopsies using an offline MudPIT shotgun proteomic approach and performed label-free quantitation through a distributed normalized spectral abundance factor approach adapted for extracted ion chromatograms (XICs). A multidimensional scaling analysis revealed that each of those tissue-types is very distinct from each other. The resection margin presented several proteins previously correlated with cancer, but also other overexpressed proteins that may be related to tumor nourishment and metastasis, such as collagen alpha-1, ceruloplasmin, calpastatin, and E-cadherin. We argue that the resection margin plays a key role in Paget's "soil to seed" hypothesis, that is, that cancer cells require a special microenvironment to nourish and that understanding it could ultimately lead to more effective treatments.

Caracterização molecular e fenotípica de estirpes de Escherichia coli produtoras de shiga-toxina (STEC) não-O157 de fezes e carcaças bovinas / Molecular and phenotypic characterization of shiga toxin producing Escherichia coli (STEC) non-O157 strains from bovine feces and carcass

Foram coletados 100 suabes retais e 100 suabes de carcaças bovinas em matadouros do estado de São Paulo, e um total de 326 estirpes de E. coli foram identificadas, sendo 163 de amostras retais e 163 de amostras de carcaça. Todos os isolados submetidos à PCR para detecção dos genes das toxinas Stx1 e Stx2 foram identificados como não-O157 e fenotipados pelo teste da citotoxicidade em células Vero. Das 26 estirpes que apresentaram apenas o gene stx1, das 56 que apresentaram apenas o gene stx2 e das 30 estirpes que apresentaram ambos os genes, 17 (65,4%), 42 (75%) e 22 (73,3%), respectivamente, foram positivas ao teste de citotoxicidade. Não houve diferença estatística entre os três perfis genéticos e na positividade ao teste de citotoxicidade. Os resultados mostram a alta frequência de expressão dos fatores de virulência das STEC de bovinos.

In the present study 100 rectal and 100 carcass swabs were collected from bovines at slaughterhouses in São Paulo state, and the total of 326 E. coli strains were identified (163 from rectal samples and 163 from carcass samples). All the isolates were submitted to PCR for Stx1 and Stx2 toxin gene detection and all strains were identified as non-O157 and phenotyped by the citotoxicity test in Vero cells. Out of 26 strains that presented only the stx1 gene, 56 that presented only the stx2 gene and 30 that presented both genes, 17 (65.4%), 42 (75%) and 22 (73.3%), respectively, were positive for the citotoxicity test. There was no statistically significant difference among these three toxinotyping profiles and positivity in the citotoxicity test, but the results show high frequency of virulence factor expression of bovine.