PARP-1 Regulates Metastatic Melanoma through Modulation of Vimentin-induced Malignant Transformation

PARP inhibition can induce anti-neoplastic effects when used as monotherapy or in combination with chemo- or radiotherapy in various tumor settings; however, the basis for the anti-metastasic activities resulting from PARP inhibition remains unknown. PARP inhibitors may also act as modulators of tumor angiogenesis. Proteomic analysis of endothelial cells revealed that vimentin, an intermediary filament involved in angiogenesis and a specific hallmark of EndoMT (endothelial to mesenchymal transition) transformation, was down-regulated following loss of PARP-1 function in endothelial cells. VE-cadherin, an endothelial marker of vascular normalization, was up-regulated in HUVEC treated with PARP inhibitors or following PARP-1 silencing; vimentin over-expression was sufficient to drive to an EndoMT phenotype. In melanoma cells, PARP inhibition reduced pro-metastatic markers, including vasculogenic mimicry. We also demonstrated that vimentin expression was sufficient to induce increased mesenchymal/pro-metastasic phenotypic changes in melanoma cells, including ILK/GSK3-β-dependent E-cadherin down-regulation, Snail1 activation and increased cell motility and migration. In a murine model of metastatic melanoma, PARP inhibition counteracted the ability of melanoma cells to metastasize to the lung. These results suggest that inhibition of PARP interferes with key metastasis-promoting processes, leading to suppression of invasion and colonization of distal organs by aggressive metastatic cells.     

Distribution in tissue homogenates,and the nature of the linkage of injected proteins to subcellular particles

Intravenously injected labeled proteins were recovered mostly in particulate fractions of rat liver homogenate. Distribution showed changes depending on the time elapsed from the injection. ¹³¹I-albumin undergoes an intraparticulate hydrolysis which shows the highest activity in the gradient fractions associated with the highest level of acid phosphatase. The labeled albumin-bearing particles separated at 27,000 g × 10 minutes released their radioactive protein at the same rate as acid phosphatase appeared in the medium, under the effect of such agents as distilled water, salts, homogenization, sonication and pH changes. The substitution of sucrose for distilled water or salts showed that the particles behave as an osmotic system as do lysosomes. These experiments prove that secondary lysosomes involved in the hydrolysis of foreign proteins, whose existence was shown by other authors only at the histochemical level, may survive the distrupting action of conventional homogenization and maintain many properties characteristic of primary lysosomes in addition to the ability of hydrolysing “in vitro” the engulfed material.     

Stimulating action of methyl 12, 12, 12-trifluorofarnesoate on in vitro juvenile hormone III biosynthesis in blattella germanica

Methyl 12, 12, 12-trifluorofarnesoate (MTFF) at a dose of 10 μM, stimulated in vitro juvenile hormone (JH) release in corpora allata (CA) from 6-day-old, freshly ecdysed, and 8-day-old (period of ootheca transport) adult virgin females of Blattella germanica. In addition, MTFF also induced intraglandular accumulation of JH and MF in treated CA. Trifluorofarnesoic acid (TFFA) and trifluorofarnesol (TFF) exhibited the same properties, although to a lesser extent than MTFF. The detection of MTFF in TFFA-treated CA suggested that TFFA and TFF were biotransformed into MTFF by the CA enzymatic system and that this ester might be responsible for the activity observed. Equivalent experiments carried out with farnesoic acid (FA) resulted in a more significant stimulation of JH production. This is not surprising, because exogenous FA is readily epoxidized at C10-C11 double bond and methylated to afford JH. Conversely, analytical data have shown that the C6-C7 double bond of MTFF is epoxidized by the CA enzymatic system, whereas that at C10-C11 remains practically unaltered.     

THDP17 Decreases Ammonia Production through Glutaminase Inhibition. A New Drug for Hepatic Encephalopathy Therapy

Ammonia production is implicated in the pathogenesis of hepatic encephalopathy (HE), being intestinal glutaminase activity the main source for ammonia. Management of ammonia formation can be effective in HE treatment by lowering intestinal ammonia production. The use of glutaminase inhibitors represents one way to achieve this goal. In this work, we have performed a search for specific inhibitors that could decrease glutaminase activity by screening two different groups of compounds: i) a group integrated by a diverse, highly pure small molecule compounds derived from thiourea ranging from 200 to 800 Daltons; and ii) a group integrated by commonly use compounds in the treatment of HE. Results shown that THDP-17 (10 µM), a thiourea derivate product, could inhibit the intestinal glutaminase activity (57.4±6.7%). Inhibitory effect was tissue dependent, ranging from 40±5.5% to 80±7.8% in an uncompetitive manner, showing V_max and K_m values of 384.62 µmol min⁻¹, 13.62 mM with THDP-17 10 µM, respectively. This compound also decreased the glutaminase activity in Caco-2 cell cultures, showing a reduction of ammonia and glutamate production, compared to control cultures. Therefore, the THDP-17 compound could be a good candidate for HE management, by lowering ammonia production.     

Neurotrophic Effects of Extracellular Guanosine

Central nervous system (CNS) astrocytes release guanosine extracellularly, that exerts trophic effects. In CNS, extracellular guanosine (GUO) stimulates mitosis, synthesis of trophic factors, and cell differentiation, including neuritogenesis, is neuroprotective, and reduces apoptosis due to several stimuli. Specific receptor-like binding sites for eGUO in the nervous system may mediate its effects through both MAP kinase and PI3-kinase signalling pathways. Extracellular guanine (eGUA) also exerts several effects; the trophic effects of eGUO are likely regulated by conversion of eGUO to eGUA by a membrane located purine nucleoside phosphorylase (ecto-PNP) and by conversion of eGUA to xanthine by guanine deaminase.     

A New Species of the Genus Paravelia Breddin 1898 from Argentina (Heteroptera: Veliidae)

A new species of the genus Paravelia (Heteroptera: Veliidae) is described, increasing the number of species of the genus known from Argentina to three: P. platensis Berg 1883, P. paxila Drake 1957, and the new sp. P. correntina. Some ecological and statistical notes are included. Scanning electron micrographs and illustrations are provided.     

Resistance of the AF-28 sorghum variety to Contarinia sorghicola

The variety AF-28 of Sorghum bicolor Pers and the hybrid R-1090 were compared in relation to the sorghum midge, Contarinia sorghicola (Coquillet), (Diptera: Cecidomyidae) in two counties of the State of São Paulo. They were compared in seven successive monthly trials beginning in September 1972. Averages of 252 and 137 adult flies emerged per head of the hybrid R-1090 in Campinas and Jaboticabal respectively, while 2.9 and 0.8 emerged from AF-28. The damage was graded with a visual scale from 0 to 4. Average damage grades of 3.5 and 3.3 were attributed to the hybrid R-1090 in Campinas and Jaboticabal respectively, while the variety AF-28 had averages of 0.5 and 0.3, respectively.

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Résumé La variété AF-28 et l'hybride R-1090 de Sorghum bicolor Pers. ont été comparés à différentes époques de l'année et en deux localités différentes de l'Etat de Sâo Paulo, Brésil.On n'a pas constaté d'interaction variété x époque ou variété x lieu.AF-28 a prouvé sa résistance à toutes les époques, aussi bien dans un lieu que dans l'autre; l'hybride par contre s'est montré grandement susceptible.Des moyennes de 252,2 et 136,7 mouches par panicule ont été observées pour l'hybride R-1090 à Campinas et à Jaboticabal contre 2,9 et 0,8 seulement par panicule de AF-28. Le préjudice a été estimé visuellement selon une échelle de 0 à 4. Des préjudices d'une valeur moyenne de 3,5 et 3,3 ont été attribués à l'hybride R-1090 à Campinas et à Jaboticabal respectivement, alors que simultanément la variété AF-28 ne subissait que des préjudices évalués à 0,5 et 0,3 respectivement.Quatre espèces de microhyménoptères parasites de Contarinia sorghicola ont été récoltées au cours de ce travail: Eupelmus popa et trois espèces différentes du genre Tetrastichus.

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Part of a thesis presented to the School of Sciences of Bauru for fulfilment of the requirement for the degree of Doctor of Science. This research was financed by the Project 72/1471 of the agreement UNIÃO-ESTADO-FAPESP and by CNPq Project 12910/73.