Cyclic AMP and calcium interplay as second messengers in melatonin-dependent regulation of Plasmodium falciparum cell cycle

The host hormone melatonin increases cytoplasmic Ca2+ concentration and synchronizes Plasmodium cell cycle (Hotta, C.T., M.L. Gazarini, F.H. Beraldo, F.P. Varotti, C. Lopes, R.P. Markus, T. Pozzan, and C.R. Garcia. 2000. Nat. Cell Biol. 2:466–468). Here we show that in Plasmodium falciparum melatonin induces an increase in cyclic AMP (cAMP) levels and cAMP-dependent protein kinase (PKA) activity (40 and 50%, respectively)

Utilização do processamento digital de imagens no monitoramento de estações de tratamento de efluentes

Neste trabalho é proposto um procedimento para reconhecimento semi-automático das espécies de protozoários por análise de imagem. Este programa (ProtoRec v.2) foi em seguida utilizado para estudar a evolução da microfauna durante períodos de transição (paragem e arranque da estação). Os resultados obtidos foram satisfatórios em relação ao reconhecimento das espécies pelo programa e o acompanhamento da estação permitiu-nos verificar a degradação e envelhecimento da lama e o aparecimento de protozoários que evidenciam tal fato

The P450 oxidoreductase, RedA, controls development beyond the mound stage in Dictyostelium discoideum

<p>Abstract</p> <p>Background</p> <p>NADPH-cytochrome-P450 oxidoreductase (CPR) is a ubiquitous enzyme that belongs to a family of diflavin oxidoreductases and is required for activity of the microsomal cytochrome-P450 monooxygenase system. CPR gene-disruption experiments have demonstrated that absence of this enzyme causes developmental defects both in mouse and insect.</p> <p>Results</p> <p>Annotation of the sequenced genome of <it>D. discoideum </it>revealed the presence of three genes (<it>redA</it>, <it>redB </it>and <it>redC</it>) that encode putative members of the diflavin oxidoreductase protein family. <it>redA </it>transcripts are present during growth and early development but then decline, reaching undetectable levels after the mound stage. <it>redB </it>transcripts are present in the same levels during growth and development while <it>redC </it>expression was detected only in vegetative growing cells. We isolated a mutant strain of <it>Dictyostelium discoideum </it>following restriction enzyme-mediated integration (REMI) mutagenesis in which <it>redA </it>was disrupted. This mutant develops only to the mound stage and accumulates a bright yellow pigment. The mound-arrest phenotype is cell-autonomous suggesting that the defect occurs within the cells rather than in intercellular signaling.</p> <p>Conclusion</p> <p>The developmental arrest due to disruption of <it>redA </it>implicates CPR in the metabolism of compounds that control cell differentiation.</p

Effects of camptothecin derivatives and topoisomerase dual inhibitors on Trypanosoma cruzi growth and ultrastructure

BACKGROUND: Trypanosoma cruzi is the etiological agent of Chagas’ disease that is an endemic disease in Latin America and affects about 8 million people. This parasite belongs to the Trypanosomatidae family which contains a single mitochondrion with an enlarged region, named kinetoplast that harbors the mitochondrial DNA (kDNA). The kinetoplast and the nucleus present a great variety of essential enzymes involved in DNA replication and topology, including DNA topoisomerases. Such enzymes are considered to be promising molecular targets for cancer treatment and for antiparasitic chemotherapy. In this work, the proliferation and ultrastructure of T. cruzi epimastigotes were evaluated after treatment with eukaryotic topoisomerase I inhibitors, such as topotecan and irinotecan, as well as with dual inhibitors (compounds that block eukaryotic topoisomerase I and topoisomerase II activities), such as baicalein, luteolin and evodiamine. Previous studies have shown that such inhibitors were able to block the growth of tumor cells, however most of them have never been tested on trypanosomatids. RESULTS: Considering the effects of topoisomerase I inhibitors, our results showed that topotecan decreased cell proliferation and caused unpacking of nuclear heterochromatin, however none of these alterations were observed after treatment with irinotecan. The dual inhibitors baicalein and evodiamine decreased cell growth; however the nuclear and kinetoplast ultrastructures were not affected. CONCLUSIONS: Taken together, our data showed that camptothecin is more efficient than its derivatives in decreasing T. cruzi proliferation. Furthermore, we conclude that drugs pertaining to a certain class of topoisomerase inhibitors may present different efficiencies as chemotherapeutical agents

WNK2 inhibits autophagic flux in human glioblastoma cell line

The following are available online at http://www.mdpi.com/2073-4409/9/2/485/s1, Figure S1: Validation of WNK2 overexpression by RT-PCR.Autophagy is a cell-survival pathway with dual role in tumorigenesis, promoting either tumor survival or tumor death. WNK2 gene, a member of the WNK (with no lysine (K)) subfamily, acts as a tumor suppressor gene in gliomas, regulating cell migration and invasion; however, its role in autophagy process is poorly explored. The WNK2-methylated human glioblastoma cell line A172 WT (wild type) was compared to transfected clones A172 EV (empty vector), and A172 WNK2 (WNK2 overexpression) for the evaluation of autophagy using an inhibitor (bafilomycin A1—baf A1) and an inducer (everolimus) of autophagic flux. Western blot and immunofluorescence approaches were used to monitor autophagic markers, LC3A/B and SQSTM1/p62. A172 WNK2 cells presented a significant decrease in LC3B and p62 protein levels, and in LC3A/B ratio when compared with control cells, after treatment with baf A1 + everolimus, suggesting that WNK2 overexpression inhibits the autophagic flux in gliomas. The mTOR pathway was also evaluated under the same conditions, and the observed results suggest that the inhibition of autophagy mediated by WNK2 occurs through a mTOR-independent pathway. In conclusion, the evaluation of the autophagic process demonstrated that WNK2 inhibits the autophagic flux in glioblastoma cell line.This project was supported by the Barretos Cancer Hospital Internal Research Funds (PAIP) to Rui Manuel Reis and by the Public Ministry of Labor Campinas (Research, Prevention, and Education of Occupational Cancer Project), Campinas, Brazil. Ana Laura Vieira Alves is the recipient of a FAPESP master fellowship (2016/18907-0)

Euglossine bees (Hymenoptera, Apidae) in a remnant of Atlantic Forest

ABSTRACT. Species composition, relative abundance, seasonal changes in the species abundance and scent association of male Euglossini collected in a semi-deciduous forest fragment in the north of the State of Paraná, southern Brazil, were recorded. Euglossine males were collected twice a month, for twelve months, from 8:00 am to 3:00 pm. The scents eucalyptol, eugenol, vanillin, methyl salicylate and benzyl acetate were used as baits. A total of 434 males distributed among 3 genera and 9 species were attracted to the chemical baits. Cockerell, 1904 (0.23%) and Eufriesea auriceps Friese, 1899 (0.23%). In general, bees were more abundant in warm-wet season (September-March). Eufriesea violacea was the most seasonal species, showing activity through the warm-wet season, from October to February. Eucalyptol was the most attractive fragrance, which was responsible for 92.6% of all visits by euglossine bees

Contaminación por Staphylococcus aureus en el procesamiento de un embutido fermentado português (linguiça)

Linguiça is a Portuguese dry-fermented sausage, which has been found to harbour food-borne pathogens in the past. Hence, the objective of this study was to investigate the levels of total viable counts (TVC), Enterobacteriaceae, and S. aureus at the key production stages of linguiça by depicting their changes using principal component analysis. Unlike Enterobacteriaceae counts, which decreased from raw meat to final product, S. aureus increased significantly in the meats throughout processing. While Enterobacteriaceae was very sensitive to the decrease in water activity, S. aureus remained viable and developed during fermentation. The presence of S. aureus at all stages should prompt industries to reinforce good hygiene practices in the processing of linguiça.Esta investigación se realizó dentro del proyecto PTDC/AGR-TEC/3107/2012, financiado por la Fundación Portuguesa de Ciencia y Tecnología (FCT)/Fondos Europeos de Desarrollo Regional (FEDER). La Dra. Gonzales-Barron agradece el apoyo financiero provisto por la FCT a través del programa "Investigator Fellowship" (IF/00570)info:eu-repo/semantics/publishedVersio

Valorization of Phenolic and Carotenoid Compounds of Sechium edule (Jacq. Swartz) Leaves: Comparison between Conventional, Ultrasound- and Microwave-Assisted Extraction Approaches

Chayote leaves are known for culinary and traditional medicine applications. This work intended to recover carotenoids and phenolic compounds from chayote leaves using the ultrasound-assisted extraction (UAE). A Box–Behnken design was employed to investigate the impact of extraction time, temperature, and ultrasonic power on the recovery of total carotenoids, total phenolic compounds, and antioxidant activities. For comparative purposes, chayote leaf extracts were prepared by maceration (ME) and microwave-assisted extraction (MAE), using the same time and temperature conditions optimized by UAE. Extraction at 50 °C and 170 Watts for 30 min provided the optimal UAE conditions. UAE showed better extraction efficacy than ME and MAE. The HPLC analysis of the extracts showed that the xanthophyll class was the main class of carotenoids, which constituted 42–85% of the total carotenoid content, followed by β-carotene and tocopherol. Moreover, 26 compounds, classified as phenolic acids, flavonols, flavonoids and other polar compounds, were identified in the chayote leaf extracts. Flavonols accounted for 55% of the total compounds quantified (the major compound was myricetin) and phenolic acids represented around 35%, mostly represented by ferulic acid, chlorogenic acid and (+)-catechin. This study revealed the potential of UAE as an effective green extraction technique to recover bioactive compounds from chayote leaves, for food, and for pharmaceutical and cosmetic applicationsThis work was supported through the projects UIDB/50006/2020 and UIDP/50006/2020, funded by FCT/MCTES through national funds. Elsa F. Vieira and Manuela Moreira thank FCT (Fundação para a Ciência e Tecnologia) for funding through the Scientific Employment Stimulus-Individual Call (Ref. CEECIND/03988/2018 and CEEC- IND/02702/2017, respectively). The authors also thank the project SYSTEMIC “an integrated approach to the challenge of sustainable food systems: adaptive and mitigatory strategies to address climate change and malnutrition”. The Knowledge hub on Nutrition and Food Security has received funding from national research funding parties in Belgium (FWO), France (INRA), Germany (BLE), Italy (MIPAAF), Latvia (IZM), Norway (RCN), Portugal (FCT), and Spain (AEI) in a joint action of JPI HDHL, JPI-OCEANS and FACCE-JPI, launched in 2019, under the ERA-NET ERA-HDHL (no 696295)info:eu-repo/semantics/publishedVersio