Interaction of a potential antitumoral benzothieno[3,2-b]pyrrole with lipid membranes and salmon sperm DNA

This work was funded by FCT and FEDER through CQ-UM and CFUM, project POCI/QUI/59407/2004 and post-Doc grant (SFRH/BPD/24548/2005) of A.S.A

Role of protein kinase R in the killing of Leishmania major by macrophages in response to neutrophil elastase and TLR4 via TNF and IFN

In cutaneous leishmaniasis, Leishmania amazonensis activates macrophage double-stranded, RNA-activated protein kinase R (PKR) to promote parasite growth. In our study, Leishmania major grew normally in RAW cells, RAW-expressing dominant-negative PKR (PKR-DN) cells, and macrophages of PKR-knockout mice, revealing that PKR is dispensable for L. major growth in macrophages. PKR activation in infected macrophages with poly I:C resulted in parasite death. Fifty percent of L. major-knockout lines for the ecotin-like serine peptidase inhibitor (ISP2; Δisp2/isp3), an inhibitor of neutrophil elastase (NE), died in RAW cells or macrophages from 129Sv mice, as a result of PKR activation. Inhibition of PKR or NE or neutralization of Toll-like receptor 4 or 2(TLR4 or TLR2) prevented the death of Δisp2/isp3. Δisp2/isp3 grew normally in RAW-PKR-DN cells or macrophages from 129Sv pkr−/−, tlr2−/−, trif−/−, and myd88−/− mice, associating NE activity, PKR, and TLR responses with parasite death. Δisp2/isp3 increased the expression of mRNA for TNF-α by 2-fold and of interferon β (IFNβ) in a PKR-dependent manner. Antibodies to TNF-α reversed the 95% killing by Δisp2/isp3, whereas they grew normally in macrophages from IFN receptor–knockout mice. We propose that ISP2 prevents the activation of PKR via an NE-TLR4-TLR2 axis to control innate responses that contribute to the killing of L. major.—Faria, M. S., Calegari-Silva, T. C., de Carvalho Vivarini, A., Mottram, J. C., Lopes, U. G., Lima, A. P. C. A. Role of protein kinase R in the killing of Leishmania major by macrophages in response to neutrophil elastase and TLR4 via TNFα and IFNβ

Fluorescence studies on potential antitumoral heteroaryl and heteroannulated indoles in solution and in lipid membranes

Fluorescence properties of three potential antitumoral compounds, a 3-(dibenzothien-4-yl)indole 1, a phenylbenzothienoindole 2 and a 3-(dibenzofur-4-yl)indole 3, were studied in solution and in lipid aggregates of dipalmitoyl phosphatidylcholine (DPPC), dioleoyl phosphatidylethanolamine (DOPE) and egg yolk phosphatidylcholine (Egg-PC). The 3-(dibenzofur-4-yl)indole 3 exhibits the higher fluorescence quantum yields in all solvents studied (0.32 ≤ ΦF ≤ 0.51). All the compounds present a solvent sensitive emission, with significant red shifts in alcohols. The results point to an ICT character of the excited state, more pronounced for compound 1. Fluorescence (steady-state) anisotropy measurements of the compounds incorporated in lipid aggregates of DPPC, DOPE and Egg-PC indicate that the three compounds are deeply located in the lipid bilayer, feeling the difference between the rigid gel phase and fluid phases.Fundação para a Ciência e a Tecnologia (FCT) - Projecto POCI/QUI/59407/2004, bolsa doutoramento SFRH/BPD/24548/2005Fundo Europeu de Desenvolvimento Regional (FEDER

Fluorescence studies of new potential antitumoral indole derivatives in lipid membranes

This work was funded by FCT and FEDER through CFUM and CQ-UM, project POCI/QUI/59407/2004 and post-Doc grant (SFRH/BPD/24548/2005) of A.S.

Embryonal rhabdomyosarcoma of the uterine cervix: a rare case report

Embryonal rhabdomyosarcoma of the female genital tract is a rare tumour. It tends to occur during childhood in the vagina and, rarely, it can arise in the uterine cervix, with a peak incidence in the second decade. We report a case of a 18-year-old female with an embryonal rhabdomyosarcoma (sarcoma botryoides) presenting himself as a cervical polyp. This tumour consisted of rhabdomyoblasts with miscellaneous differentiation surrounded by a loose, myxoid stroma. The patient was successfully treated with cervical conization and adjuvant chemotherapy. She is now disease-free at the 28th month follow-up. Awareness of this uncommon lesion in the cervix and its clinical implications is important to prevent misdiagnosis. Therapy has recently inclined to conservative and fertility-sparing treatment

Blood flow of bubbles moving in microchannels with bifurcations

The gas embolism is a well-known phenomenon. Previous studies have been performed to understand the formation, the behavior and the influence of air bubbles in microcirculation. This study aims to investigate the flow of bubbles in a microchannel network with bifurcations. For that purpose, a microchannel network was fabricated by soft lithography. The working fluids used were composed by sheep red blood cells (RBCs) suspended in dextran 40 and two different hematocrits were studied, 5% and 10%. The in vitro blood flow was analyzed for a flow rate of lO ~-tllmin, by using an inverted microscope and a high-speed camera. It was possible to visualize the formation of the bubbles and their behavior along the network. The results show that the passage of air bubbles influences the cells local concentration, since a higher concentration of cells was seen upstream to the bubble and lower concentrations downstream to the bubble.This work was supported by Fundação para a Ciência e a Tecnologia (FCT), Portugal, under the strategic grants UID/EMS/04077/2019, UID/EEA/04436/2019 and UID/EMS/00532/2019. The authors are also grateful for the partial funding of FCT through the projects POCI-01-0145-FEDER-016861 (ref: PTDC/QEQ-FIT/4287/2014), NORTE-01-0145- FEDER-029394, NORTE-01-0145-FEDER-030171, funded by COMPETE2020, NOR1E2020, PORTUGAL2020, and FEDER. D. Bento acknowledges the PhD scholarship SFRH/BD/911921 2012 granted by FCT.info:eu-repo/semantics/publishedVersio

New tetracyclic heteroaromatic compounds based on dehydroamino acids : photophysical and electrochemical studies of interaction with DNA

A benzothienoindole (BTIN) and a benzofuroindole (BFIN) were synthesized in high yields, as potential new target DNA compounds, using a metal-assisted intramolecular C-N cyclization, developed by us, of the methyl esters of N-(t-butoxycarbonyl)-b,b-bis(dibenzothien-4-yl or dibenzofuro-4-yl)dehydroalanines. The latter were obtained by a bis-Suzuki coupling of a b,b-dibromodehydroalanine with the corresponding heteroarylboronic acids. The absorption and fluorescence properties of the novel tetracyclic heteroaromatic compounds were studied in different solvents and in the presence of natural double-stranded (ds) salmon sperm DNA. The results in several solvents show that either BTIN or BFIN can be used as fluorescence solvent sensitive probes. Spectroscopic studies of the interaction of both compounds with dsDNA allowed to determine binding constant (Ki) values and binding site sizes (n). Fluorescence quenching experiments using iodide ion allowed the determination of the accessibilities to the quencher, showing that intercalation is the preferred mode of binding of these molecules to DNA. From the results obtained BTIN is the more intercalative compound and has a higher affinity to DNA. The interaction of this more promising compound with DNA was also studied electrochemically, by using differential pulse voltammetry (DPV) in connection with disposable pencil graphite electrode (PGE). These studies are based on the differences in the BTIN and adenine oxidation signals. After the interaction of BTIN with DNA, the oxidation signals of BTIN and adenine strongly decreased. The latter was attributed to the binding of the BTIN to DNA and the former points to a possible damage of the oxidizable groups of the compound after intercalation into DNA. Several concentrations of BTIN were tested and 50 μg/mL was found to be the optimum concentration in order to detect its interaction with DNA. In addition, the detection limit and the reproducibility were determined by using a disposable electrochemical transducer. The results of spectroscopic and electrochemical detection of BTIN interaction with DNA are in good agreement.Academy of Pharmacists and Turkish Pharmacists Association (TEB)Turkish Academy of Sciences - Young Scientist Award Program (KAE/TUBA-GEBIP/2001-2-8)FEDERFundação para a Ciência e a Tecnologia (FCT) - Projecto POCI/QUI/59407/2004. A.S.A. acknowledges a post-doc. grant SFRH/BPD/24548/2005, Bolsa SFRH/BPD/24548/2005.Scientific and Technical Research Council of Turkey (TUBITAK)