Identification of Tsetse (Glossina spp.) using matrix-assisted laser desorption/ionisation time of flight mass spectrometry

Glossina (G.) spp. (Diptera: Glossinidae), known as tsetse flies, are vectors of African trypanosomes that cause sleeping sickness in humans and nagana in domestic livestock. Knowledge on tsetse distribution and accurate species identification help identify potential vector intervention sites. Morphological species identification of tsetse is challenging and sometimes not accurate. The matrix-assisted laser desorption/ionisation time of flight mass spectrometry (MALDI TOF MS) technique, already standardised for microbial identification, could become a standard method for tsetse fly diagnostics. Therefore, a unique spectra reference database was created for five lab-reared species of riverine-, savannah- and forest- type tsetse flies and incorporated with the commercial Biotyper 3.0 database. The standard formic acid/acetonitrile extraction of male and female whole insects and their body parts (head, thorax, abdomen, wings and legs) was used to obtain the flies' proteins. The computed composite correlation index and cluster analysis revealed the suitability of any tsetse body part for a rapid taxonomical identification. Phyloproteomic analysis revealed that the peak patterns of G. brevipalpis differed greatly from the other tsetse. This outcome was comparable to previous theories that they might be considered as a sister group to other tsetse spp. Freshly extracted samples were found to be matched at the species level. However, sex differentiation proved to be less reliable. Similarly processed samples of the common house fly Musca domestica (Diptera: Muscidae; strain: Lei) did not yield any match with the tsetse reference database. The inclusion of additional strains of morphologically defined wild caught flies of known origin and the availability of large-scale mass spectrometry data could facilitate rapid tsetse species identification in the futur

An integrated omics analysis reveals molecular mechanisms that are associated with differences in seed oil content between Glycine max and Brassica napus

Abstract Background: Rapeseed (Brassica napus L.) and soybean (Glycine max L.) seeds are rich in both protein and oil, which are major sources of biofuels and nutrition. Although the difference in seed oil content between soybean (~ 20%) and rapeseed (~ 40%) exists, little is known about its underlying molecular mechanism. Results: An integrated omics analysis was performed in soybean, rapeseed, Arabidopsis (Arabidopsis thaliana L. Heynh), and sesame (Sesamum indicum L.), based on Arabidopsis acyl-lipid metabolism- and carbon metabolism-related genes. As a result, candidate genes and their transcription factors and microRNAs, along with phylogenetic analysis and co-expression network analysis of the PEPC gene family, were found to be largely associated with the difference between the two species. First, three soybean genes (Glyma.13G148600, Glyma.13G207900 and Glyma.12G122900) co-expressed with GmPEPC1 are specifically enriched during seed storage protein accumulation stages, while the expression of BnPEPC1 is putatively inhibited by bna-miR169, and two genes BnSTKA and BnCKII are co-expressed with BnPEPC1 and are specifically associated with plant circadian rhythm, which are related to seed oil biosynthesis. Then, in de novo fatty acid synthesis there are rapeseed-specific genes encoding subunits β-CT (BnaC05g37990D) and BCCP1 (BnaA03g06000D) of heterogeneous ACCase, which could interfere with synthesis rate, and β-CT is positively regulated by four transcription factors (BnaA01g37250D, BnaA02g26190D, BnaC01g01040D and BnaC07g21470D). In triglyceride synthesis, GmLPAAT2 is putatively inhibited by three miRNAs (gma-miR171, gma-miR1516 and gma-miR5775). Finally, in rapeseed there was evidence for the expansion of gene families, CALO, OBO and STERO, related to lipid storage, and the contraction of gene families, LOX, LAH and HSI2, related to oil degradation. Conclusions: The molecular mechanisms associated with differences in seed oil content provide the basis for future breeding efforts to improve seed oil content

Expression and pharmacological inhibition of TrkB and EGFR in glioblastoma

A member of the Trk family of neurotrophin receptors, tropomyosin receptor kinase B (TrkB, encoded by the NTRK2 gene) is an increasingly important target in various cancer types, including glioblastoma (GBM). EGFR is among the most frequently altered oncogenes in GBM, and EGFR inhibition has been tested as an experimental therapy. Functional interactions between EGFR and TrkB have been demonstrated. In the present study, we investigated the role of TrkB and EGFR, and their interactions, in GBM. Analyses of NTRK2 and EGFR gene expression from The Cancer Genome Atlas (TCGA) datasets showed an increase in NTRK2 expression in the proneural subtype of GBM, and a strong correlation between NTRK2 and EGFR expression in glioma CpG island methylator phenotype (G-CIMP+) samples. We showed that when TrkB and EGFR inhibitors were combined, the inhibitory effect on A172 human GBM cells was more pronounced than when either inhibitor was given alone. When U87MG GBM cells were xenografted into the flank of nude mice, tumor growth was delayed by treatment with TrkB and EGFR inhibitors, given alone or combined, only at specific time points. Intracranial GBM growth in mice was not significantly affected by drug treatments. Our findings indicate that correlations between NTRK2 and EGFR expression occur in specific GBM subgroups. Also, our results using cultured cells suggest for the first time the potential of combining TrkB and EGFR inhibition for the treatment of GBM

Collection and morphological characterization of sweet potato landraces in north of Rio de Janeiro state Coleta e caracterização morfológica de variedades locais de batata-doce no norte do Rio de Janeiro

The traditional farmers play an important role in plant genetic resources conservation. Collecting the germplasm maintained by these farmers is a very important action to avoid genetic variability losses. The goals of this work were to collect sweet potato from farms in the north of Rio de Janeiro state; to gather information regarding to the farmers profile, and to characterize the sweet potato landraces collected using morphological descriptors. Fifty three farms were visited in six collection expedition and 46 accessions were collected. During the visits the farmers were interviewed using a query with ten items. Six root traits and eight descriptors for vegetative parts were used for morphological characterization. The data were analyzed based on Cole-Rodgers distance and clustering was done with UPGMA method. Familiar agriculture with subsistence objective was observed and sweet potato was cultivated by 72% of the farmers at least for more than a decade, supporting the observation that this vegetable is traditionally cultivated in small areas in the specific region. The morphological characterization was efficient to detect genetic variability among accessions, revealing that traditional farmers from Campos dos Goytacazes and São João da Barra are responsible for sweet potato genotypes conservation with expressive genetic diversity in their properties. There was no relationship between genetic distance and collecting areas.<br>Os agricultores tradicionais têm um papel fundamental na conservação dos recursos genéticos vegetais e a coleta de germoplasma mantido por esses produtores é muito importante para evitar a perda da variabilidade genética. Os objetivos deste trabalho foram coletar germoplasma de batata-doce em propriedades rurais situadas no norte do estado do Rio de Janeiro; levantar informações quanto ao perfil dos produtores rurais visitados durante as coletas, e caracterizar morfologicamente as variedades locais de batata-doce coletadas. Em seis viagens de coleta foram visitadas 53 propriedades rurais, e coletados 46 acessos. Em todas as visitas foi possível entrevistar o produtor por meio de questionário contendo dez itens. Para a caracterização, foram usados seis caracteres de raiz e oito da parte aérea. A análise dos dados foi efetuada com o uso da distância de Cole-Rodgers e o agrupamento foi gerado com o método UPGMA. Foi constatado predomínio da agricultura familiar, voltada para subsistência, sendo que 72% dos agricultores cultivavam a batata-doce há mais de uma década, o que evidencia a tradição de cultivo desta hortaliça em pequenas áreas na região. A caracterização morfológica foi eficiente para detectar a variabilidade genética entre os acessos, demonstrando que os agricultores tradicionais dos municípios de Campos dos Goytacazes e São João da Barra detêm genótipos de batata-doce com expressiva diversidade genética em suas propriedades. Não foi detectada correlação entre a distância genética e os locais de coleta