Isolation of flavonoids from Tagetes patula (French Marigold) flowers: Cytotoxic and oxidant activity in human cervical carcinoma cells / Aislación de flavonoides de las flores de Tagetes patula (cempasúchil francés): Actividad citotóxica y oxidante en células de carcinoma cervical humano

T agetes patula , known as French Marigold, belongs to the family Asteraceae. Human papillomavirus infection is considered one of the causes of cervical cancer. This study assessed the cytotoxic activity and intracellular oxidative capacity of compounds isolated from extract of T. patula flowers as anti - cancer cervical agents. Fraction F6 of n - butanol extract was subjected to column chromatography and HPLC - ESI - MS. The isolated compo unds of T. patula were used to examine cytotoxic activity and the production of total reactive oxygen species in SiHa and HeLa cells; the cells were also characterized using scanning electron microscopy. Patulitrin was cytotoxic to SiHa and HeLa cells. An increase in ROS production was observed at different times of treatment of cells with patuletin and patulitrin. Scanning electron microscopy showed morphological changes in SiHa and HeLa cells. Thus, compounds isolated from T. patula have great treatment p otential against cervical cancer.

Tagetes patula , conocida como cempasúchil francés, pertenece a la familia Asteraceae. La infección por el virus del papiloma humano se considera una de las causas del cáncer cervical. En este estudio, se evaluó la actividad citotóxica y la capacidad oxidativa intracelular de los compuestos aislados del extracto de las flores de T. patula como agentes anticancerígenos cervicales. La fracción F6 del ext racto de n - butanol se sometió a cromatografía en columna y HPLC - ESI - MS. Los compuestos aislados de T. patula se utilizaron para examinar la actividad citotóxica y la producción total de especies reactivas de oxígeno en las células SiHa y HeLa; las células también se caracterizaron mediante microscopía electrónica de barrido. Patulitrina resultó citotóxica para las células SiHa y HeLa. Se observó un aumento en la producción de ROS en diferentes momentos del tratamiento de las células con patuletina y patulit rina. La microscopía electrónica de barrido mostró cambios morfológicos en las células SiHa y HeLa. Por lo tanto, los compuestos aislados de T. patula tienen un gran potencial de tratamiento contra el cáncer cervical.

Quinoxalines against Leishmania amazonensis: SAR study, proposition of a new derivative, QSAR prediction, synthesis, and biological evaluation.

Neglected tropical diseases, such as leishmaniasis, lead to serious limitations to the affected societies. In this work, a structure-activity relationship (SAR) study was developed with a series of quinoxaline derivatives, active against the promastigote forms of Leishmania amazonensis. As a result, a new quinoxaline derivative was designed and synthesized. In addition, a quantitative structure-activity relationship (QSAR) model was obtained [pIC50 = - 1.51 - 0.96 (EHOMO) + 0.02 (PSA); N = 17, R2 = 0.980, R2Adj = 0.977, s = 0.103, and LOO-cv-R2 (Q2) = 0.971]. The activity of the new synthesized compound was estimated (pIC50 = 5.88) and compared with the experimental result (pIC50 = 5.70), which allowed to evaluate the good predictive capacity of the model.

Antiproliferative Activity and Ultrastructural Changes in Promastigote and Amastigote forms of Leishmania amazonensis Caused by Limonene-Acylthiosemicarbazide Hybrids.

Leishmaniasis is a tropical zoonotic disease. It is found in 98 countries, with an estimated 1.3 million people being affected annually. During the life cycle, the Leishmania parasite alternates between promastigote and amastigote forms. The first line treatment for leishmaniasis are the pentavalent antimonials, such as N-methylglucamine antimoniate (Glucantime®) and sodium stibogluconate (Pentostam®). These drugs are commonly related to be associated with dangerous side effects such as cardiotoxicity, nephrotoxicity, hepatotoxicity, and pancreatitis. Considering these aspects, this work aimed to obtain a new series of limonene-acylthiosemicarbazides hybrids as an alternative for the treatment of leishmaniasis. For this, promastigotes, axenic amastigotes, and intracellular amastigotes of Leishmania amazonensis were used in the antiproliferative assay; J774-A1 macrophages for the cytotoxicity assay; and electron microscopy techniques were performed to analyze the morphology and ultrastructure of parasites. ATZ-S-04 compound showed the best result in both tests. Its IC50 , in promastigotes, axenic amastigotes and intracellular amastigotes was 0.35±0.08 µM, 0.49±0.06 µM, and 15.90±2.88 µM, respectively. Cytotoxicity assay determined a CC50 of 16.10±1.76 µM for the same compound. By electron microscopy, it was observed that ATZ-S-04 affected mainly the Golgi complex, in addition to morphological changes in promastigote forms of L. amazonensis.

Photophysical characterization of Hypericin-loaded in micellar, liposomal and copolymer-lipid nanostructures based F127 and DPPC liposomes.

Hypericin (Hy) compound presents a high photoactivity in photodynamic therapy (PDT), photodiagnosis and theranostics applications. The maintenance of this compound in monomeric form could undermine the potential benefits of its photophysical and photodynamic activity. In this study, we demonstrated that the Hy formulated in a system based on the use of the F127 copolymer and the 1,2-dipalmitoyl-sn-3-glycerol-phosphatidylcholine (DPPC) as micelles, liposomal vesicles and Copolymer-Lipid coated systems, have improved its photophysical properties for many clinical modalities. Based on the results of the triplet state lifetime values (τt), the singlet oxygen quantum yield (ΦΔ1O2), the fluorescence lifetime (τF) and the fluorescence quantum yield (ΦF), all Hy formulations had its photophysical properties described in different models of drug delivery systems (DDS). In addition, the transient spectra profile of those formulations was unaffected by the Hy incorporation process, except for the liposomal system, which demonstrated to be the less stable one by flash photolysis technique. The cytotoxic effects of those formulations were also investigated for CaCo-2 and HaCat cells line. The cytotoxic concentrations for 50% (CC50) were 0.56, 1.05, 1.33 and 4.80 µmol L-1 for Copolymer-Lipid/Hy, DPPC/Hy, F127/Hy and ethanol/Hy for CaCo-2 cells, respectively, and 0.69, 2.02, 1.45 and 1.16 µmol L-1 for Copolymer-Lipid/Hy, DPPC/Hy, F127/Hy and ethanol/Hy for HaCat cells, respectively. The F127 copolymer had a significant role in many photophysical parameters determined for Copolymer-Lipid/Hy coated system. Although all those formulations had shown satisfactory results, Copolymer-Lipid/Hy proved to be superior in many aspects, being the most promising formulation for PDT, photodiagnosis and theranostics applications.

A versatile method for the selective core-crosslinking of hyaluronic acid nanogels via ketone-hydrazide chemistry: from chemical characterization to in vivo biodistribution.

The development of biopolymer-based nanogels has gained particular interest to achieve successful delivery of therapeutics for the treatment of various diseases, such as cancer, infection and diabetes. Herein, we report a new and simple methodology for the covalent stabilization of self-assembled gel nanoparticles based on hyaluronic acid (HA) modified with a thermoresponsive ketone-functional copolymer. This relies on the selective formation of hydrazone crosslinks with bishydrazides within the globular domains of the copolymer chains formed above the cloud point temperature. This approach allows tuning of the crosslinking density by varying the dihydrazide crosslinker to ketone molar ratio. The size distributions and morphology of the nanogels were assessed using dynamic light scattering (DLS), cryo-transmission and scanning electron microscopy. In vitro cellular uptake in several cancer cells and in vivo biodistribution of the nanogels in different mouse tumor models were then explored to assess the effectiveness of this crosslinking strategy. The data from these experiments show prolonged blood circulation, longer than 24 hours, for the crosslinked nanogels and high tumor accumulation.

The Combination of Vitamin K3 and Vitamin C Has Synergic Activity against Forms of Trypanosoma cruzi through a Redox Imbalance Process.

Chagas' disease is an infection that is caused by the protozoan Trypanosoma cruzi, affecting millions of people worldwide. Because of severe side effects and variable efficacy, the current treatments for Chagas' disease are unsatisfactory, making the search for new chemotherapeutic agents essential. Previous studies have reported various biological activities of naphthoquinones, such as the trypanocidal and antitumor activity of vitamin K3. The combination of this vitamin with vitamin C exerted better effects against various cancer cells than when used alone. These effects have been attributed to an increase in reactive oxygen species generation. In the present study, we evaluated the activity of vitamin K3 and vitamin C, alone and in combination, against T. cruzi. The vitamin K3 + vitamin C combination exerted synergistic effects against three forms of T. cruzi, leading to morphological, ultrastructural, and functional changes by producing reactive species, decreasing reduced thiol groups, altering the cell cycle, causing lipid peroxidation, and forming autophagic vacuoles. Our hypothesis is that the vitamin K3 + vitamin C combination induces oxidative imbalance in T. cruzi, probably started by a redox cycling process that leads to parasite cell death.

Susceptibilidade a antifúngicos e produção de enzimas por leveduras do (levaduras del) gênero Candida isoladas (aisladas) de pacientes com HIV/AIDS / Susceptibility to antifungal and enzyme production by Candida yeasts isolated from HIV/AIDS patients

A candidíase é a (constituye la) infecção fúngica mais frequente entre os pacientes portadores do vírus da imunodeficiência humana. Este estudo objetivou o isolamento (el aislamiento), identificação, avaliaçãodo (evaluación del) perfil de susceptibilidade e fatores de virulência de leveduras do gênero Candida provenientes de pacientes HIV/AIDS. As amostras foram coletadas (Las muestras se recolectaron) de 100 pacientes em diferentes sítios anatômicos (boca, trato gastrointestinal e vaginal), cultivadas em meio (en medio) ágar Sabouraud, e identificadas através da cultura em meio (mediante cultivo en medio) diferencial CHRO Mágar Candida e pela técnica da reação em cadeia da (y por la técnica de reacción en cadena de la) polimerase (PCR). O teste de (La prueba de) susceptibilidade in vitro aos (a los) antifúngicos fluconazol, anfotericia B e nistatina foram determinados através da técnica de microdiluição em caldo (en caldo). A infectividade foi avaliada através da (se validó por medio de la) produção de fosfolipase se hemolisinas. Do total (De un total) de 45 leveduras isoladas, houve maior (hubo mayor) prevalência de C. albicans, seguida por C. glabrata, C.parapisilosis, C.tropicalis e C. krusei. Foram submetidas ao (Sesometieron a la) teste de suscptibilidade 43 (95%) das a mostras, onde a (en la que la) concentração inibitória mínima foi determinada. Todos os isolados foram sensíveis a (Todos los aislamientos eransensibles a) anfotericina B e a nistatina, en quanto 60.6% foram sensíveis ao fluconazol, 6.9% sensíveisdose-dependente e 32.5% apresentaram resistência ao (presentaron resistencia al) fluconazol. As espé-cies C. glabrata e C. tropicalis apresentaram CIMs maior para o fluconazol do que os encontrados para C.albicans. Apresentou produção de fosfolipase 62% das amostras e atividade hemolítica 87%...

Adherence of Pseudomonas aeruginosa and Candida albicans to urinary catheters

Se investigó la interacción entre aislamientos clínicos de Pseudomonasaeruginosa y Candida albicans, y su capacidad de adhesión in vitro encatéteres urinarios. Fue realizado análisis cuantitativo a través del cómputo deunidades formadoras de colonias y microscopía electrónica de barrido.Los resultados mostraron que el desarrollo de P. aeruginosa sobre catéteresurinarios fue mayor con la presencia de C. albicans, mientras que la presenciade la levadura no fue afectada significativamente. Son necesarios másestudios para comprender mejor la patogénesis de estos microorganismos ypara establecer estrategias para el control y el tratamiento de las infeccionesurinarias vinculadas al uso de catéteres(AU)

In this study, the in vitro adherence capabilities of Pseudomonas aeruginosaand Candida albicans clinical isolates to urinary catheters were investigated.Quantitative analysis was performed by colony-forming unit counts andscanning electron microscopy. Results demonstrated that the adherence ofP. aeruginosa to urinary catheters was enhanced in the presence ofC. albicans, while C. albicans adherence was not significantly affected.Further investigations are warranted to fully understand the pathogenicpotential of their interaction in order to aid in the design of novel strategies forthe prevention and treatment of catheter-related UTIs(AU)

Occurrence of Pseudomonas aeruginosain drinking water - DOI: 10.4025/actascibiolsci.v28i1.1053 / Ocorrência de Pseudomonas aeruginosa em água potável - DOI: 10.4025/actascibiolsci.v28i1.1053

A total of 413 samples of drinking water, taken from principal and secondary water systems in the municipality of Bandeirantes, state of Paraná, southern Brazil, were analyzed through membrane filtration for P. aeruginosa and on Plate Count agar for heterotrophic bacteria. Total and fecal coliforms were determined by multiple dilution tube methods. Turbidity, pH, chlorine, fluoride, alkalinity, color, dissolved oxygen, hardness and organic matter were also measured. Of the 413 samples examined, 10.41% were contaminated by P. aeruginosa, including 23,53% of the water from secondary water system, 8,56% of the water from principal water system. No coliform was found in any of the samples. One sample counted above the maximum heterotrophic bacteria level permitted. P. aeruginosa isolated from the water samples were tested for chlorine sensitivity and survived up to three times the minimum chlorine concentration used. It is desirable that Pseudomonas enumeration be performed periodically, in addition to data routinely collected by most water supply systems

Foram analisadas 413 amostras de água potável do sistema principal e do sistema secundário em Bandeirantes, Estado do Paraná, através de membrana filtrante para P. aeruginosa e bactérias heterotróficas em ágar Plate Count. Coliformes totais e fecais foram determinados pelo método dos tubos múltiplos. A qualidade físico-química da água foi determinada em termos de turbidez, pH, cloro, flúor, alcalinidade, cor, oxigênio dissolvido, dureza e matéria orgânica. Das amostras examinadas, 10,41% estavam contaminadas por P. aeruginosa, incluindo 23,53% das amostras do sistema de água secundário, e 8,56% das amostras do sistema de água principal. Somente uma amostra apresentou contagem de bactérias heterotróficas acima do nível máximo recomendado. P. aeruginosa isoladas foram testadas para sensibilidade ao cloro livre e sobreviveram a uma concentração de cloro três vezes acima da concentração mínima usada. É recomendada a determinação periódica de Pseudomonas em adição aos dados rotineiramente coletados na maioria dos sistemas de abastecimento

Bacterial microflora in the gastrointestinal tract of Nile tilapia, Oreochromis niloticus, cultured in a semi-intensive system- DOI: 10.4025/actascibiolsci.v25i2.2007 / Flora bacteriana de tilápia do Nilo, Oreochromis niloticus, cultivada em sistema semi-intensivo - DOI: 10.4025/actascibiolsci.v25i2.2007

This experiment measured total bacterial numbers in the gastrointestinal regions of semi-intensively cultured tilapia, Oreochromis niloticus L. (Cichlidae). Mean bacterial numbers were higher in both anterior and posterior gut than in stomach. The percentage of isolated species and the percentage of isolates from any particular species varied significantly among gastrointestinal tract regions. Aeromonas hydrophila, Aeromonas veronii, Burkholderia cepacia, Chromobacterium violaceum, Citrobacter freundii, Escherichia coli, Flavimonas oryzihabitans and Plesiomonas shigelloides were the most frequently isolated Gram-negative bacilli. From these samples, only Plesiomonas shigelloides was present in both gastrointestinal regions, with higher number in the posterior gut (76%), when compared to anterior gut (4.8%) and stomach (0.6%). Aeromonas hydrophila (0.6%), Escherichia coli (7.4%), and Flavimonas oryzihabitans were isolated only in the stomach, and Citrobacter freundii and Burkholderia cepacia were found only in the posterior gut. Chromobacterium violaceum was the dominant bacteria isolated from stomach and anterior gut, with 90% and 55%, respectively. Unidentified organisms comprised 0 - 39.3% of the gastrointestinal microflora

A flora bacteriana de diferentes partes do trato gastrintestinal de tilápia Oreochomis niloticus L. (Cichlidae) foi determinada. O número médio de bactérias foi maior no intestino anterior e posterior quando comparado ao estômago. A porcentagem total de espécies bacterianas isoladas e a porcentagem de espécies isoladas em uma espécie particular variaram significativamente entre as regiões do trato gastrintestinal. Aeromonas hydrophila, Aeromonas veronii, Burkholderia cepacia, Chromobacterium violaceum, Citrobacter freundii, Escherichia coli, Flavimonas oryzihabitans e Plesiomonas shigelloides foram os bacilos Gram-negativos encontrados com maior freqüência. Destas espécies, somente Plesiomonas shigelloides esteve presente em cada região do trato gastrintestinal, apresentando maior número no intestino posterior (76%), quando comparado com o intestino anterior (4.8%) e o estômago (0.6%). Aeromonas hydrophila (0.6%), Escherichia coli (7.4%), e Flavimonas oryzihabitans foram isoladas somente do estômago, e Citrobacter freundii e Burkholderia cepacia foram encontradas somente no intestino posterior. Chromobacterium violaceum foi a espécie dominante isolada do estômago e do intestino anterior com 90% e 55%, respectivamente. Organismos não identificados compreendem 0 39.3% da microbiota gastrintestinal

Haematological and biochemical values for Nile tilapia Oreochromis niloticus cultured in semi-intensive system - DOI: 10.4025/actascibiolsci.v25i2.2028 / Valores hematológicos e bioquímicos em tilápia do Nilo Oreochromis niloticus cultivada em sistema semi-intensivo - DOI: 10.4025/actascibiolsci.v25i2.2028

The aim of the present study was to investigate the reference ranges for haematological and biochemical values for Nile tilapia, Oreochromis niloticus (Cichlidae), cultivated in a semi-intensive system. Erythrocytes number was 6.93 ± 8.28 x 106/mm3, hemoglobin 10.52 ± 3.09g/dL of blood and hematocrit 31.85 ± 8.45%, mean corpuscular volume (MVC) 148.80 ±. 153.19µ3, mean corpuscular hemoglobin (MCH) 40.74 ± 34.19 pg and mean corpuscular hemoglobin concentration (MCHC) 35.24 ± 14.92%. Total plasma protein and glucose values obtained were 3.06 ± 0.65g/dl and 60.32 ± 20.22mg/dl, respectively. A correlation matrix was established to compare de degree of association among biometric data and hematological parameters, and among each of them and biochemical values. A positive correlation was observed among weight, length, MCV and MHC, as well as between length and hematocrit. The blood glucose level was positively correlated with weight and length, whereas total protein was positively correlated with hemoglobin. The erythrocytes count was positively correlated with hemoglobin and negatively correlated with MCV and MHC. Microscopy examination of blood smears revealed the presence of erythrocytes, neutrophils, lymphocytes, and monocytes. Under the conditions employed here, no basophils or eosinophils, nor their precursors, could be found in blood smears of Nile tilapia

O objetivo do presente estudo foi investigar os valores de referências hematológicos e bioquímicos para a tilápia do Nilo, Oreochromis niloticus (Cichlidae), cultivada em sistema semi-intensivo. A contagem de eritrócitos foi 6,93 ± 8,28 x 106/mm3, hemoglobina 10,52 ± 3,09g/dL de sangue, hematócrito 31,85 ± 8,45%, volume corpuscular médio (MVC) 148,80 ±.153,19µ3, hemoglobina corpuscular média (MCH) 40,74 ± 34,19pg e concentração de hemoglobina corpuscular média 35,24 ± 14,92%. Os valores de proteína plasmática total e da glicose obtidos foram de 3,06 ± 0,65g/dl e 60,32 ± 20,22mg/dl, respectivamente. Uma matriz de correlação foi estabelecida para comparar o grau de associação entre os dados biométricos e parâmetros hematológicos, e entre estes e os valores bioquímicos. Uma correlação positiva foi observada entre peso, comprimento, MCV e MHC, bem como entre comprimento e hematócrito. O nível de glicose sanguínea foi positivamente correlacionado com o peso e o comprimento, enquanto proteína total foi positivamente correlacionada com a hemoglobina. A contagem de eritrócito foi positivamente correlacionada com a hemoglobina e negativamente correlacionada com MCH e MHC. A observação microscópica do esfregaço sangüíneo revelou a presença de eritrócitos, neutrófilos, linfócitos e monócitos. Sob as condições empregadas no estudo não foram encontrados basófilos, ensinófilos ou seus precur

Enumeration and isolation of facultative anaerobic bacteria in an upflow anaerobic sludge blanket reactor treating wastewater from a gelatine industry- DOI: 10.4025/actascibiolsci.v25i2.1977 / Enumeração e isolamento de bactérias anaeróbias facultativas num reator de fluxo ascendente e manta de lodo tratando efluente de uma indústria de gelatina - DOI: 10.4025/actascibiolsci.v25i2.1977

This experiment measured levels of facultative anaerobic bacteria in an Upflow Anaerobic Sludge Blanket (UASB) reactor treating wastewater from a gelatin industry. The microorganism quantification showed similar values in granular sludge bed and fluidized zone. In the settling compartment of reactor, the bacteria were two logs lower. These communities are composed mainly of Gram-negative rods. The most abundant genera isolated were Acinetobacter, Aeromonas, Citrobacter, Escherichia, Klebsiella and Stenotrophomonas. The significance of the bacteria isolated from UASB reactor for public health is unknown. Some species are permanent residents; other are found only in a fraction of the population, and still other species are opportunistic pathogens that cause human infections. In the present study, none of the predominant bacteria belonged to the group of strict pathogens

Neste trabalho foram determinados os níveis de bactérias anaeróbias facultativas em Reator de Fluxo Ascendente e Manta de Lodo (UASB) utilizado no tratamento de efluente de industria de gelatina. A quantificação dos microrganismos apresentou similar valor na manta de lodo granular e na zona de fluidização. No compartimento de sedimentação do reator as bactérias foram dois logs menores. Estas comunidades são compostas principalmente de bacilos Gram-negativos. Os mais abundantes gêneros foram Acinetobacter, Aeromonas, Citrobacter, Escherichia, Klebsiella e Stenotrophomonas. O significado para a saúde pública das bactérias isoladas no reator UASB é desconhecido. Algumas espécies são residentes permanentes, outras são encontradas em somente uma parte da população, e ainda outras espécies são patógenos oportunistas que causam infecção humana. No presente estudo, nenhuma das bactérias predominantes pertence ao grupo dos patógenos estritos