LaSOM 335, active against bladder cancer cells, interferes with Let-60 (hRas) and reduces CD73 expression/activity.

Bladder cancer is the fourth most common malignancy in men. It can present along the entire continuum of severity, from mild to well-differentiated disease to extremely malignant tumors with low survival rates. Human RAS genes are the most frequently mutated oncogenes in human cancers, and the critical role of aberrant Ras protein function in carcinogenesis is well established. Therefore, considerable efforts have been devoted to the development of anti-Ras inhibitors for cancer treatment. This study presents the biphenyl dihydropyrimidinone LaSOM 335 with high activity against T24 bladder cancer cells (IC50 = 10.73 ± 0.53 µM) and selectivity of cytotoxicity for this cancer cell line compared to two non-cancer cell lines investigated. Furthermore, we also show that this compound reduced vulvar development in the mutant let-60 gene of Caenorhabditis elegans. Let-60 is a homolog of the mammalian Ras gene. In addition, we observed that LaSOM 335 inhibits the enzymatic activity of CD73 and decreases CD73 expression. Possibly, this expression decrease is due to downstream EGFR signaling via the Ras-Raf-ERK pathway, that directly regulates CD73 expression via ERK1/2. Evidence suggests that non-immunomodulating functions of CD73 play an equally important role for cancer cell survival, progression, and migration. Regarding we also notice that LaSOM 335 was safe in the in vivo model of C. elegans. The set of these findings makes this biphenyl dihydropyrimidinone a promising candidate for further investigations in the bladder cancer field.

Seroprevalence and systemic immune biomarkers associated with Toxoplasma gondii infection in blood donors from Southern Brazil.

The high seroprevalence of Toxoplasma gondii infection in Blood Banks could be a potential risk for contamination of blood recipients. The discovery of new biomarkers may help to distinguish between seropositive and seronegative donors. This study determined the seroprevalence and profile of systemic immune biomarkers associated with Toxoplasma gondii infection among blood donors from Southern Brazil. Peripheral blood was collected from 510 blood donors (52.2 % male; mean age: 36.61), 310, and 200 from Erechim, and Chapecó municipalities, respectively. Specific Toxoplasma gondii IgG and IgM antibodies were detected by Eletrochemioluminescence. Nested PCR and qPCR were performed to detectToxoplasma gondii DNA. Twenty-seven inflammatory factors were analyzed using a high-performance Luminex assay. Among 310 blood donors from Erechim, 44.5 % (138/310) were IgM(-)/IgG(+), and 1.3 % (4/310) were IgM(+)/IgG(+), while out of 200 blood donors from Chapeco, 42.5 % (85/200) were IgM(-)/IgG(+), and 2 % (4/200) were IgM(+)/ IgG(+). We did not find Toxoplasma gondii DNA in the samples analyzed by Nested PCR and qPCR.Additionally, IgM(-)/IgG(+) donors presented higher levels ofdistinct systemic mediators, and were indicated to be high producers of several systemic mediators (CCL11, CCL2, CCL3, CCL4, CXCL10, IL-1ß, IL-17, IFN-γ, IL-4, IL-9, IL-13, IL-10, IL-1Ra, vascular endothelial growth factor/VEGF, platelet-derived growth factor/PDGF, granulocyte-macrophage colony-stimulating factor/GM-CSF, and IL-7). However, IgM(+)/IgG(+) donors were found as high producers of CXCL8, CXCL10, CCL4, IL-1ß, IL-1Ra, IL-9, IL-13, and PDGF, while IgM(-)/IgG(-) donors showed unaltered levels for the most soluble mediators evaluated. These distinct biomarker signatures might help identify potential factors to distinguish between IgM(-) and IgM(+) donors.

The Role of Alternative Toxicological Trials in Drug Discovery Programs. The Case of Caenorhabditis elegans and Other Methods.

The discovery of a new drug requires over a billion dollars and around 12 years of research efforts, and toxicity is the leading reason for the failure to approve candidate drugs. Many alternative methods have been validated to detect toxicity as early as possible to diminish the waste of resources and efforts in medicinal chemistry research, and in vivo alternative methods are especially valuable for the amount of information they can provide at little cost and in a short time. In this work, we present a review of the literature published between the years 2000 and 2021 on in vivo alternative methods of toxicity screening employed in medicinal chemistry, which we believe will be useful because, in addition to shortening the research time, these studies provide much additional information aside from the toxicity of drug candidate compounds. These in vivo models include zebrafish, Artemia salina, Galleria mellonella, Drosophila melanogaster, planarians, and Caenorhabditis elegans. The most published ones in the last decade were zebrafish, D. melanogaster, and C. elegans due to their reliability, ease, and cost-effectiveness in implementation and flexibility. Special attention is given to C. elegans because of its rising popularity, a wide range of uses, including toxicity screening, and active effects measurement, from antioxidant effects to anthelmintic and antimicrobial activities, and its fast and reliable results. Over time, C. elegans also became a viable high-throughput (HTS) automated drug screening option. Additionally, this manuscript lists briefly the other screening methods used for the initial toxicological analyses and the role of alternative in vivo methods in these scenarios, classifying them as in silico, in vitro and alternative in vivo models that have been receiving a growing increase in interest in recent years.

Exploring the N1 Position of Biginelli Compounds: New Insights and Trends for Chemical Diversity Generation of Bioactive Derivatives.

Dihydropyrimidinones (DHPMs) are heterocycles obtained by the multicomponent Biginelli reaction. Recently, new synthetic protocols have allowed us to explore functionalisation at less explored positions of DHPMs, such as the N1 position. In this context, a full literature survey of N1- substituted DHPMs was performed. We analysed 27 papers and identified 379 compounds with substituents at the N1 position, most of them with alkyl groups, and a total of 28% compounds with aromatic substituents attached at the N1 position. N1-substituted DHPMs were explored mainly due to their effects on cancer cell proliferation via numerous targets, such as kinesin Eg5, heat shock protein 70, heat shock protein 90, and the epidermal growth factor receptor. Similarity analyses were performed using the data of 379 DHPMs from different cheminformatic approaches, i.e., chemical property correlations, principal component analysis, similarity networks, and compound clustering.

Discovery, development, chemical diversity and design of isoxazoline-based insecticides.

Isoxazoline is a 5-membered heterocycle present in the active compounds of many commercial veterinary anti-ectoparasitic products. The molecular target of isoxazolines is the inhibition of GABA-gated chloride channels in insects. These facts have inspired the use of the isoxazoline scaffold in the design of novel insecticide compounds. The main strategies used for isoxazoline synthesis are either the 1,3-dipolar cycloaddition between a nitrile oxide and an alkene or the reaction between hydroxylamine and an α,ß-unsaturated carbonyl compound. This review highlights the utilization of isoxazoline as insecticide: its mode of action, its commercial preparations and its consideration in the design of novel insecticides. Similarity analyses were performed with 235 isoxazoline derivatives in three different cheminformatic approaches - chemical property correlations, similarity network and compound clustering. The cheminformatic methodologies are interesting tools to use in evaluating the similarity between commercial isoxazolines and to clarify the main features explored within their derivatives.

New pharmacological findings linked to biphenyl DHPMs, kinesin Eg5 ligands: anticancer and antioxidant effects.

Background: Dihydropyrimidin-2-thiones (DHPMs) are a class of heterocyclic compound which have been intensively investigated mainly due to their anticancer activity as kinesin Eg5 inhibitors. Materials & methods: A library of N1 aryl substituted DHPMs were tested against glioma and bladder cancer cell lines. Quantitative structure-activity relationship (QSAR) investigation was performed in order to identify key elements of DHPMs linked with their antiproliferative effect. The toxicity of most active compounds was investigated using Caenorhabditis elegans as the model. Results & conclusion: DHPMs 9, 13 and 17 have been identified as having improved activity against glioma and bladder cell lines as compared with monastrol. Flow cytometry investigations showed that the new compounds induce cell cycle arrest in phase G2/M and cell death by apoptosis. In addition, compound 13 was able to modulate the reactive oxygen species production in vivo in C. elegans. The biphenyl dihydropyrimidinthiones provided a safety profile in C. elegans.

Novel coumarins active against Trypanosoma cruzi and toxicity assessment using the animal model Caenorhabditis elegans.

BACKGROUND: Chagas disease (CD) is a tropical parasitic disease. Although the number of people infected is very high, the only drugs available to treat CD, nifurtimox (Nfx) and benznidazole, are highly toxic, particularly in the chronic stage of the disease. Coumarins are a large class of compounds that display a wide range of interesting biological properties, such as antiparasitic. Hence, the aim of this work is to find a good antitrypanosomal drug with less toxicity. The use of simple organism models has become increasingly attractive for planning and simplifying efficient drug discovery. Within these models, Caenorhabditis elegans has emerged as a convenient and versatile tool with significant advantages for the toxicological potential identification for new compounds. METHODS: Trypanocidal activity: Forty-two 4-methylamino-coumarins were assayed against the epimastigote form of Trypanosoma cruzi (Tulahuen 2 strain) by inhibitory concentration 50% (IC50). Toxicity assays: Lethal dose 50% (LD50) and Body Area were determined by Caenorhabditis elegans N2 strain (wild type) after acute exposure. Structure-activity relationship: A classificatory model was built using 3D descriptors. RESULTS: Two of these coumarins demonstrated near equipotency to Nifurtimox (IC50 = 5.0 ± 1 µM), with values of: 11 h (LaSOM 266), (IC50 = 6.4 ± 1 µM) and 11 g (LaSOM 231), (IC50 = 8.2 ± 2.3 µM). In C. elegans it was possible to observe that Nfx showed greater toxicity in both the LD50 assay and the evaluation of the development of worms. It is possible to observe that the efficacy between Nfx and the synthesized compounds (11 h and 11 g) are similar. On the other hand, the toxicity of Nfx is approximately three times higher than that of the compounds. Results from the QSAR-3D study indicate that the volume and hydrophobicity of the substituents have a significant impact on the trypanocidal activities for derivatives that cause more than 50% of inhibition. These results show that the C. elegans model is efficient for screening potentially toxic compounds. CONCLUSION: Two coumarins (11 h and 11 g) showed activity against T. cruzi epimastigote similar to Nifurtimox, however with lower toxicity in both LD50 and development of C. elegans assays. These two compounds may be a feasible starting point for the development of new trypanocidal drugs.

Avaliação dos níveis séricos de colesterol total em indivíduos atendidos em um laboratório universitário / Assessment of total cholesterol levels in individuals attended in a university laboratory

Objetivo: Avaliar os níveis séricos de colesterol total em indivíduos atendidos em um laboratório universitário de análises clínicas, localizado em Erechim, RS. Métodos: Níveis de colesterol total foram obtidos a partir de uma análise retrospectiva transversal com indivíduos de ambos os sexos e idade entre 20-79 anos, entre 2010 a 2013. Resultados: Os níveis de colesterol total de 1.475 pacientes, incluindo 479 homens e 996 mulheres, foram analisados. Na população feminina, 46,89% das participantes apresentaram valores normais de colesterol total, e este índice para a população masculina correspondeu a 50,31%. Foi encontrada uma correlação significativa entre a idade e o colesterol total nos participantes do sexo feminino (p=0,019). Em ambos os sexos foi observado um aumento nos níveis de colesterol total durante as primeiras décadas de vida, seguido por estabilização a partir da quinta e sexta décadas de vida, respectivamente para homens e mulheres. Conclusão: A prevalência de hipercolesterolemia foi elevada na população do estudo, o que indica a necessidade de apropriadas estratégias de intervenção para profilaxia de risco cardiovascular

Objective: To estimate the prevalence of hypercholesterolemia amongadults attended in a university laboratory, localized in Erechim/RS. Methods: Cholesterol total levels were obtained from s transversal retrospective analysis with individuals of both genders and age between 20-79 years, from 2010 to 2013. Results: The total cholesterol levels of 1.475 patients, including 479 men and 996 women were analyzed. In female gender 46.89% of participants showed normal total cholesterol levels, and this value in masculine population corresponded to 50.31%. Was found a significantly correlation between age and total cholesterol in female participants (p=0.019). In both genders was observed a increase in total cholesterol levels during the first life decades, followed by stabilization from fifth and sixth life decade, respectively for men and women. Conclusion: The prevalence hypercholesterolemia was high in study population, which necessitates appropriate intervention strategies to prevent cardiovascular risk.

Metabolic modeling and comparative biochemistry in glyoxylate cycle / Modelagem metabólica e bioquímica comparativo no ciclo do glioxilato

Glyoxylate cycle in fatty acid catabolism enhances net production of oxaloacetate, a substrate for gluconeogenesis, in certain bacteria, invertebrates and oilseed in the growth stage. A theoretical model was developed to calculate ATP amount produced in each step of the catabolic pathway, taking into account the fatty acids hydrocarbon chain size. Results showed a decrease in energy efficiency in glyoxylate cycle when compared to animal metabolism. Although the glyoxylate cycle provides evolutionary adaptations, it determines a smaller amount of energy produced per carbon atom when compared to animal catabolism of fatty acids.(AU)

Em algumas bactérias, invertebrados e sementes em germinação, o ciclo do glioxilato no catabolismo de ácidos graxos permite a produção líquida de oxaloacetato, substrato para a gliconeogênese. Foi desenvolvido um modelo teórico para calcular a quantidade de ATP produzida em cada etapa desta rota metabólica, considerando o tamanho da cadeia hidrocarbonada do ácido graxo. Os resultados mostraram uma diminuição na eficiência energética do ciclo do glioxilato em relação ao metabolismo animal. Embora o ciclo do glioxilato confira adaptações evolutivas, ele determina uma menor quantidade de energia produzida por átomo de carbono em relação ao catabolismo animal de ácidos graxos.(AU)

Candida species distribution and fluconazole susceptibility of blood isolates at a regional hospital in Passo Fundo, RS, Brazil / Distribuição de espécies de Candida e suscetibilidade ao fluconazol de isolados de hemocultivos em hospital regional de Passo Fundo, RS, Brasil

ABSTRACT Introduction: Candidemia is a bloodstream infection produced by Candida genus yeasts. Objective: The purpose of this study was to characterize the epidemiology and the fluconazole susceptibility in Candida species isolated from patients at a regional hospital in Passo Fundo, RS. Methods: Records from the laboratory were used to identify patients with positive blood cultures for Candida between 2010 and 2011. The in vitro activity of fluconazole was determined using the disk diffusion method. Results: Were analyzed 24 positive blood cultures for Candida and found a 54.16% mortality rate. C. albicans was the most prevalent species, followed by C. parapsilosis and C. krusei. For susceptibility to fluconazole, C. albicans, C. parapsilosis and C. tropicalis showed 100% sensitivity. However, C. krusei was 100% resistant; and C. glabrata, 50% resistant. Conclusion: The high mortality and fluconazole resistance rates emphasize the importance of the diagnosis of candidemia in a hospital environment. .

RESUMO Introdução: Candidemia é a infecção da corrente sanguínea produzida por leveduras do gênero Candida. Objetivos: Este estudo teve por objetivo caracterizar a epidemiologia e o perfil de suscetibilidade ao fluconazol em espécies de Candida, isoladas em pacientes internados em um hospital regional de Passo Fundo, RS. Métodos: Registros laboratoriais foram utilizados para identificar pacientes com hemocultura positiva para espécies do gênero Candida, entre 2010 e 2011. A atividade in vitro do fluconazol foi determinada por meio do método de difusão em disco. Resultados: Foram analisadas 24 hemoculturas positivas para Candida; a taxa de mortalidade encontrada foi 54,16%. C. albicans foi a espécie mais prevalente, seguida por C. parapsilosis e C. krusei. C. albicans, C. parapsilosis e C. tropicalis apresentaram 100% de sensibilidade ao fluconazol, entretanto C. krusei mostrou-se 100% resistente; e C. glabrata, 50% resistente. Conclusão: A elevada taxa de mortalidade e resistência ao fluconazol enfatiza a importância do diagnóstico de candidemia em ambiente hospitalar. .

Total Phenolic Contents and Antioxidant Activity in Oxidized Leaves of Mate (Ilex paraguariensis St. Hil)

Ilex paraguariensis is a native species to South America. Its economic importance is in the use of its leaves and twigs in the processing of a product popularly known as "chimarrão". The aim of this study was to evaluate the correlations between some physico-chemical process parameters in the oxidation of I. paraguariensis leaves oxidation. The leaves of Maté with age 1.0, 6.5 and 12.0 months were subjected to an oxidation process for 3 h under different relative humidity conditions (80, 90 and 99%) at 26°C. The extracts were evaluated using the experimental design techniques to maximize the oxidation conditions for the total polyphenols, antioxidant capacity, water activity and color. The condition of 90% humidity and leaf age 6.5 months resulted in lower lightness and yellow color of the product. Green color was not significantly influenced by the studied parameters. The highest levels of total phenols and antioxidant potential were found when the leaf age was one month and humidity was 90%. Thus, it could be concluded that the air humidity of the chamber and leaf age influenced the oxidation process of the leaves of Maté as well as the antioxidant capacity of the extract obtained.

Physico-chemical characterization of Ilex paraguariensis St. Hil. during the maturation

In Brazil, yerba mate is consumed after processing; however, in Chile and Uruguay, the consumers prefer the cured product, which acquires a yellow color. For that yerba-mate is stored for a period of six months to one year, which increases the cost of the final product for the overseas market. This study evaluated the effect of humidity and temperature in maturation chamber on the time required for the product to get this characteristic. The changes in the color, pH, moisture and water activity were evaluated during the time of storage in different conditions of temperature and humidity. Yerba-mate subjected to higher temperature and humidity showed nearest color of the product submitted to natural storage. The loss of green color was related to the reduction in pH and increase in the moisture of the samples. The higher humidity allowed the mate to reach conditions near to market requirements abroad in approximately 60 days of maturation.

Surgical resection of a cutaneous nodule in the left foot caused by mycobacteria / Exérese cirúrgica de nódulo cutâneo no pé esquerdo causado por uma micobactéria

Nontuberculous mycobacteria are etiologic agents of opportunistic human infections. Although they usually affect superficial tissues, infections in bones and joints have been described. The contamination is associated with increased environmental exposure. With appropriate therapy, the cases usually progress to complete recovery of the patient. This study reports the case of a patient who developed a cutaneous nodule in her left foot acquired when her skin was punctured by a fish. The anatomopathological examination revealed chronic central suppurative granulomatous dermo-hypodermal inflammation. Furthermore, the screening for resistant acid-fast bacilli was positive.

Micobactérias não tuberculosas são agentes causadores de infecções oportunistas no homem. Embora, geralmente, afetem tecidos superficiais, infecções em ossos e articulações têm sido descritas. O contágio está associado ao aumento da exposição do homem ao meio ambiente. Diante da terapêutica adequada, os casos normalmente evoluem com recuperação total do indivíduo. Este estudo descreve o caso clínico de um paciente que apresentou nódulo cutâneo no pé esquerdo após traumatismo com um peixe. O exame anatomopatológico revelou inflamação crônica granulomatosa centralmente supurativa dermo-hipodérmica e a pesquisa de bacilos álcool-ácido resistentes (BAAR) foi positiva.