RESUMO
Abstract Among the plants defense mechanisms, the induction and emission of volatile organic compounds, which can be used to attract natural enemies, such predators insects. Although well studied, the induction of plant volatiles that attract natural enemies can vary according to intensity of infestation of herbivores and the species of host plant. We investigated the olfactory behavioral responses of the predatory mite Neoseiulus californicus (McGregor) (Acari: Phytoseiidae) to the volatiles of infested maize (Zea mays) plants by the two-spotted spider mite Tetranychus urticae (Koch, 1836) (Acari: Tetranychidae) in early and advanced infestations. The Bt (Viptera) maize cultivar Impact® was used for tests the behavior of N. californicus. After initial and advanced infestations, the phytophagous mites T. urticae were removed, and the plants were tested using a Y olfactometer. The following treatments were evaluated: air vs. air, uninfested plants vs. air, uninfested plants vs. plants infested with 10 females of T. urticae, uninfested plants vs. plants infested with 100 females of T. urticae, uninfested plants vs. plants infested with 200 females of T. urticae and plants infested with 10 vs. plants infested with 200 females of T. urticae. The predatory mite N. californicus did not show preference to the treatments tested, suggesting that maize plants infested by T. urticae do not induce volatiles capable of attracting the predatory mite N. californicus. We concluded that N. californicus is not attracted by maize plants infested by T. urticae.
Resumo Dentre os mecanismos de defesa de plantas, a indução e emissão de compostos orgânicos voláteis, podem ser utilizados para atrair inimigos naturais, como insetos predadores. Embora bem estudada, a indução dos voláteis de plantas que atraem inimigos naturais pode variar de acordo com a intensidade de infestação de herbívoros e a espécie de planta hospedeira. Investigamos as respostas comportamentais olfativas do ácaro predador Neoseiulus californicus (McGregor) (Acari: Phytoseiidae) aos voláteis de plantas infestadas pelo ácaro-rajado Tetranychus urticae (Koch, 1836) (Acari: Tetranychidae). A cultivar de milho Bt (Viptera) Impact® foi utilizada para testar o comportamento de N. californicus. Após infestações iniciais e avançadas, os ácaros fitófagos T. urticae foram removidos e as plantas testadas em olfatômetro Y. Os seguintes tratamentos foram avaliados: ar vs. ar, plantas não infestadas vs. ar, plantas não infestadas vs. plantas infestadas com 10 fêmeas de T. urticae, plantas não infestadas vs. plantas infestadas com 100 fêmeas de T. urticae, plantas não infestadas vs. plantas infestadas com 200 fêmeas de T. urticae e plantas infestadas com 10 vs. plantas infestadas com 200 fêmeas de T. urticae. O ácaro predador N. californicus não mostrou preferência aos tratamentos testados, sugerindo que plantas de milho infestadas por T. urticae não induzem voláteis capazes de atrair o ácaro predador N. californicus. Concluímos que N. californicus não é atraído por plantas de milho infestadas por T. urticae.
RESUMO
Among the plants defense mechanisms, the induction and emission of volatile organic compounds, which can be used to attract natural enemies, such predators insects. Although well studied, the induction of plant volatiles that attract natural enemies can vary according to intensity of infestation of herbivores and the species of host plant. We investigated the olfactory behavioral responses of the predatory mite Neoseiulus californicus (McGregor) (Acari: Phytoseiidae) to the volatiles of infested maize (Zea mays) plants by the two-spotted spider mite Tetranychus urticae (Koch, 1836) (Acari: Tetranychidae) in early and advanced infestations. The Bt (Viptera) maize cultivar Impact® was used for tests the behavior of N. californicus. After initial and advanced infestations, the phytophagous mites T. urticae were removed, and the plants were tested using a "Y" olfactometer. The following treatments were evaluated: air vs. air, uninfested plants vs. air, uninfested plants vs. plants infested with 10 females of T. urticae, uninfested plants vs. plants infested with 100 females of T. urticae, uninfested plants vs. plants infested with 200 females of T. urticae and plants infested with 10 vs. plants infested with 200 females of T. urticae. The predatory mite N. californicus did not show preference to the treatments tested, suggesting that maize plants infested by T. urticae do not induce volatiles capable of attracting the predatory mite N. californicus. We concluded that N. californicus is not attracted by maize plants infested by T. urticae.
Dentre os mecanismos de defesa de plantas, a indução e emissão de compostos orgânicos voláteis, podem ser utilizados para atrair inimigos naturais, como insetos predadores. Embora bem estudada, a indução dos voláteis de plantas que atraem inimigos naturais pode variar de acordo com a intensidade de infestação de herbívoros e a espécie de planta hospedeira. Investigamos as respostas comportamentais olfativas do ácaro predador Neoseiulus californicus (McGregor) (Acari: Phytoseiidae) aos voláteis de plantas infestadas pelo ácaro-rajado Tetranychus urticae (Koch, 1836) (Acari: Tetranychidae). A cultivar de milho Bt (Viptera) Impact® foi utilizada para testar o comportamento de N. californicus. Após infestações iniciais e avançadas, os ácaros fitófagos T. urticae foram removidos e as plantas testadas em olfatômetro "Y". Os seguintes tratamentos foram avaliados: ar vs. ar, plantas não infestadas vs. ar, plantas não infestadas vs. plantas infestadas com 10 fêmeas de T. urticae, plantas não infestadas vs. plantas infestadas com 100 fêmeas de T. urticae, plantas não infestadas vs. plantas infestadas com 200 fêmeas de T. urticae e plantas infestadas com 10 vs. plantas infestadas com 200 fêmeas de T. urticae. O ácaro predador N. californicus não mostrou preferência aos tratamentos testados, sugerindo que plantas de milho infestadas por T. urticae não induzem voláteis capazes de atrair o ácaro predador N. californicus. Concluímos que N. californicus não é atraído por plantas de milho infestadas por T. urticae.
Assuntos
Animais , Feminino , Tetranychidae , Ácaros e Carrapatos , Ácaros , Comportamento Predatório , Olfato , Zea maysRESUMO
Among the plants defense mechanisms, the induction and emission of volatile organic compounds, which can be used to attract natural enemies, such predators insects. Although well studied, the induction of plant volatiles that attract natural enemies can vary according to intensity of infestation of herbivores and the species of host plant. We investigated the olfactory behavioral responses of the predatory mite Neoseiulus californicus (McGregor) (Acari: Phytoseiidae) to the volatiles of infested maize (Zea mays) plants by the two-spotted spider mite Tetranychus urticae (Koch, 1836) (Acari: Tetranychidae) in early and advanced infestations. The Bt (Viptera) maize cultivar Impact® was used for tests the behavior of N. californicus. After initial and advanced infestations, the phytophagous mites T. urticae were removed, and the plants were tested using a "Y" olfactometer. The following treatments were evaluated: air vs. air, uninfested plants vs. air, uninfested plants vs. plants infested with 10 females of T. urticae, uninfested plants vs. plants infested with 100 females of T. urticae, uninfested plants vs. plants infested with 200 females of T. urticae and plants infested with 10 vs. plants infested with 200 females of T. urticae. The predatory mite N. californicus did not show preference to the treatments tested, suggesting that maize plants infested by T. urticae do not induce volatiles capable of attracting the predatory mite N. californicus. We concluded that N. californicus is not attracted by maize plants infested by T. urticae.
Dentre os mecanismos de defesa de plantas, a indução e emissão de compostos orgânicos voláteis, podem ser utilizados para atrair inimigos naturais, como insetos predadores. Embora bem estudada, a indução dos voláteis de plantas que atraem inimigos naturais pode variar de acordo com a intensidade de infestação de herbívoros e a espécie de planta hospedeira. Investigamos as respostas comportamentais olfativas do ácaro predador Neoseiulus californicus (McGregor) (Acari: Phytoseiidae) aos voláteis de plantas infestadas pelo ácaro-rajado Tetranychus urticae (Koch, 1836) (Acari: Tetranychidae). A cultivar de milho Bt (Viptera) Impact® foi utilizada para testar o comportamento de N. californicus. Após infestações iniciais e avançadas, os ácaros fitófagos T. urticae foram removidos e as plantas testadas em olfatômetro Y. Os seguintes tratamentos foram avaliados: ar vs. ar, plantas não infestadas vs. ar, plantas não infestadas vs. plantas infestadas com 10 fêmeas de T. urticae, plantas não infestadas vs. plantas infestadas com 100 fêmeas de T. urticae, plantas não infestadas vs. plantas infestadas com 200 fêmeas de T. urticae e plantas infestadas com 10 vs. plantas infestadas com 200 fêmeas de T. urticae. O ácaro predador N. californicus não mostrou preferência aos tratamentos testados, sugerindo que plantas de milho infestadas por T. urticae não induzem voláteis capazes de atrair o ácaro predador N. californicus. Concluímos que N. californicus não é atraído por plantas de milho infestadas por T. urticae.
Assuntos
Animais , Controle Biológico de Vetores/métodos , Volatilização , Ácaros e Carrapatos/parasitologia , Zea maysRESUMO
Among the plants defense mechanisms, the induction and emission of volatile organic compounds, which can be used to attract natural enemies, such predators insects. Although well studied, the induction of plant volatiles that attract natural enemies can vary according to intensity of infestation of herbivores and the species of host plant. We investigated the olfactory behavioral responses of the predatory mite Neoseiulus californicus (McGregor) (Acari: Phytoseiidae) to the volatiles of infested maize (Zea mays) plants by the two-spotted spider mite Tetranychus urticae (Koch, 1836) (Acari: Tetranychidae) in early and advanced infestations. The Bt (Viptera) maize cultivar Impact® was used for tests the behavior of N. californicus. After initial and advanced infestations, the phytophagous mites T. urticae were removed, and the plants were tested using a "Y" olfactometer. The following treatments were evaluated: air vs. air, uninfested plants vs. air, uninfested plants vs. plants infested with 10 females of T. urticae, uninfested plants vs. plants infested with 100 females of T. urticae, uninfested plants vs. plants infested with 200 females of T. urticae and plants infested with 10 vs. plants infested with 200 females of T. urticae. The predatory mite N. californicus did not show preference to the treatments tested, suggesting that maize plants infested by T. urticae do not induce volatiles capable of attracting the predatory mite N. californicus. We concluded that N. californicus is not attracted by maize plants infested by T. urticae.
Dentre os mecanismos de defesa de plantas, a indução e emissão de compostos orgânicos voláteis, podem ser utilizados para atrair inimigos naturais, como insetos predadores. Embora bem estudada, a indução dos voláteis de plantas que atraem inimigos naturais pode variar de acordo com a intensidade de infestação de herbívoros e a espécie de planta hospedeira. Investigamos as respostas comportamentais olfativas do ácaro predador Neoseiulus californicus (McGregor) (Acari: Phytoseiidae) aos voláteis de plantas infestadas pelo ácaro-rajado Tetranychus urticae (Koch, 1836) (Acari: Tetranychidae). A cultivar de milho Bt (Viptera) Impact® foi utilizada para testar o comportamento de N. californicus. Após infestações iniciais e avançadas, os ácaros fitófagos T. urticae foram removidos e as plantas testadas em olfatômetro Y. Os seguintes tratamentos foram avaliados: ar vs. ar, plantas não infestadas vs. ar, plantas não infestadas vs. plantas infestadas com 10 fêmeas de T. urticae, plantas não infestadas vs. plantas infestadas com 100 fêmeas de T. urticae, plantas não infestadas vs. plantas infestadas com 200 fêmeas de T. urticae e plantas infestadas com 10 vs. plantas infestadas com 200 fêmeas de T. urticae. O ácaro predador N. californicus não mostrou preferência aos tratamentos testados, sugerindo que plantas de milho infestadas por T. urticae não induzem voláteis capazes de atrair o ácaro predador N. californicus. Concluímos que N. californicus não é atraído por plantas de milho infestadas por T. urticae.(AU)
Assuntos
Animais , Ácaros e Carrapatos/parasitologia , Controle Biológico de Vetores/métodos , Volatilização , Zea maysRESUMO
We use optical tweezers to characterize the ability of Caffeine (Caf) to modulate the intercalation of drugs into the DNA double-helix at the single molecule level. When previously bound to the double-helix, Caf hinders ethidium bromide (EtBr) intercalation, decreasing its effective equilibrium binding constant with DNA. The dominant mechanism of such singular ability is a direct binding of Caf to the intercalating drugs in solution, which decreases the effective concentration of such compounds available to interact with DNA. When EtBr intercalation into the DNA double-helix occurs firstly, on the other hand, the measured cooperativity between Caf molecules interacting with DNA can be modulated, a feature also correlated to the Caf-EtBr interaction in solution. The results achieved here unveil many peculiarities about the details of such interactions at the molecular level and provide new insights on the use of Caf in therapeutic applications.
Assuntos
Substâncias Intercalantes , Cafeína , DNA/química , Conformação de Ácido NucleicoRESUMO
Plants not only respond to herbivorous damage but adjust their defense system after egg deposition by pest insects. Thereby, parasitoids use oviposition-induced plant volatiles to locate their hosts. We investigated the olfactory behavioral responses of Trichogramma pretiosum Riley, 1879 (Hymenoptera: Trichogrammatidae) to volatile blends emitted by maize (Zea mays L.) with singular and stacked events after oviposition by Spodoptera frugiperda Smith, 1797 (Hymenoptera: Trichogrammatidae) moths. Additionally, we examined possible variations in gene expression and on oviposition-induced volatiles. We used a Y-tube olfactometer to test for the wasp responses to volatiles released by maize plants oviposited by S. frugiperda and not-oviposited plants. Using the real-time PCR technique (qRT-PCR), we analyzed the expression of lipoxygenase and three terpene synthases genes, which are enzymes involved in the synthesis of volatile compounds that attract parasitoids of S. frugiperda. Olfactometer tests showed that T. pretiosum is strongly attracted by volatiles from transgenic maize emitted by S. frugiperda oviposition (VTPRO 3, more than 75% individuals were attracted). The relative expression of genes TPS10, LOX e STC was higher in transgenic hybrids than in the conventional (isogenic line) hybrids. The GC-MS analysis revealed that some volatile compounds are released exclusively by transgenic maize. This study provides evidence that transgenic hybrids enhanced chemical cues under oviposition-induction and helped to increase T. pretiosum efficiency in S. frugiperda control. This finding shows that among the evaluated hybrids, genetically modified hybrids can improve the biological control programs, since they potentialize the egg parasitoid foraging, integrating pest management.
RESUMO
Among the plants defense mechanisms, the induction and emission of volatile organic compounds, which can be used to attract natural enemies, such predators insects. Although well studied, the induction of plant volatiles that attract natural enemies can vary according to intensity of infestation of herbivores and the species of host plant. We investigated the olfactory behavioral responses of the predatory mite Neoseiulus californicus (McGregor) (Acari: Phytoseiidae) to the volatiles of infested maize (Zea mays) plants by the two-spotted spider mite Tetranychus urticae (Koch, 1836) (Acari: Tetranychidae) in early and advanced infestations. The Bt (Viptera) maize cultivar Impact® was used for tests the behavior of N. californicus. After initial and advanced infestations, the phytophagous mites T. urticae were removed, and the plants were tested using a "Y" olfactometer. The following treatments were evaluated: air vs. air, uninfested plants vs. air, uninfested plants vs. plants infested with 10 females of T. urticae, uninfested plants vs. plants infested with 100 females of T. urticae, uninfested plants vs. plants infested with 200 females of T. urticae and plants infested with 10 vs. plants infested with 200 females of T. urticae. The predatory mite N. californicus did not show preference to the treatments tested, suggesting that maize plants infested by T. urticae do not induce volatiles capable of attracting the predatory mite N. californicus. We concluded that N. californicus is not attracted by maize plants infested by T. urticae.
Assuntos
Ácaros e Carrapatos , Ácaros , Tetranychidae , Animais , Feminino , Comportamento Predatório , Olfato , Zea maysRESUMO
In the past months, the use of the drug hydroxychloroquine has considerably increased in many countries, associated with a proposed treatment for the COVID-19 disease. Although there is no conclusive evidence about the efficacy of the drug for this purpose, surprisingly there are no conclusive studies in the literature concerning its mechanism of action inside cells, which is related to its interaction with nucleic acids. Here, we performed a robust characterization of the interaction between hydroxychloroquine and double-stranded DNA using single-molecule force spectroscopy and gel electrophoresis. Two different binding modes were identified, namely, minor groove binding for low drug concentrations and intercalation for high drug concentrations, and the sets of binding parameters were determined for each of these modes. Such results have unraveled in detail the molecular mechanism of action of the drug as a DNA ligand.
Assuntos
DNA/química , Hidroxicloroquina/química , Substâncias Intercalantes/química , Eletroforese , Pinças Ópticas , Imagem Individual de MoléculaRESUMO
We present a new study on the interaction of the DNA molecule with the surfactant dodecyltrimethylammonium bromide (DTAB), performed mainly with optical tweezers. Single-molecule force spectroscopy experiments performed in the low-force entropic regime allowed a robust characterization of the DNA-DTAB interaction, unveiling how the surfactant changes the mechanical properties of the biopolymer, the binding parameters, and the competition of the two mechanisms involved in the interaction: electrostatic attraction between the cationic surfactant heads and the negative phosphate backbone of the DNA and hydrophobic interactions between the tails of the bound DTAB molecules, which can result in DNA compaction in solution depending on the quantity of bound surfactant. Finally, force clamp experiments with magnetic tweezers and gel electrophoresis assays confirm that DTAB compacts DNA depending not only on the surfactant concentration but also on the conformation of the biopolymer in solution. The present study provides new insights on general aspects of the DNA-surfactant complexes formation, contributing to the fundamental knowledge of the physics of such interactions.
Assuntos
DNA/química , DNA/metabolismo , Interações Hidrofóbicas e Hidrofílicas , Compostos de Amônio Quaternário/farmacologia , Eletricidade Estática , Tensoativos/farmacologia , Modelos Moleculares , Conformação de Ácido Nucleico , Pinças Ópticas , Compostos de Amônio Quaternário/química , Compostos de Amônio Quaternário/metabolismo , Tensoativos/química , Tensoativos/metabolismoRESUMO
Europium oxide (Eu2O3) was used to evaluate the affinity of this rare earth element for interacting with double-stranded (ds) DNA molecules. To perform the study, we used single molecule force spectroscopy with optical tweezers and gel electrophoresis assays. Force spectroscopy experiments show that Eu2O3 presents a strong interaction with dsDNA, and the binding is independent on the ionic strength used in the surrounding environment. Among the main characteristics of the interaction, Eu2O3 tends to bind in a cooperative way, forming bound clusters of â¼ 3 molecules, and presents a high equilibrium association binding constant on the order of 105 M-1. In addition, gel electrophoresis confirm the weak electrostatic character of the interaction and explicit show that Eu2O3 does not interfere on drug intercalation into the double-helix. Such results demonstrate the potential of europium for interacting with nucleic acids and strongly suggest that this rare earth element may be considered for the design of new metal-based anticancer drugs in the future.
Assuntos
Antineoplásicos/farmacologia , Desenho de Fármacos , Európio/farmacologia , DNA/química , Modelos Moleculares , Imagem Individual de MoléculaRESUMO
By performing single-molecule force spectroscopy with optical tweezers, we have characterized the interaction between the platinum-based compound transplatin and the DNA molecule, establishing a critical comparison with its isomer cisplatin. While transplatin is ineffective against tumor cells, its isomer is one of the most used drugs in current chemotherapies, and a molecular study on this difference performed at the single-molecule level was lacking until the present work. Our experiments show that transplatin binds DNA under low chloride concentrations (a situation usually found inside many cells) with an equilibrium association binding constant about four orders of magnitude lower than cisplatin. In addition, we have found that, at saturation, transplatin binds preferentially forming interstrand cross links and monoadducts, a situation very different from cisplatin, which forms preferentially intrastrand cross links. Such differences explain the ineffectiveness of transplatin in killing tumor cells. From a physical point of view, the present study advances in using the mechanical properties of the DNA molecule as sensors to evaluate the therapeutic efficiency of drugs.
Assuntos
Antineoplásicos/farmacologia , Cisplatino/farmacologia , Imagem Individual de Molécula , Antineoplásicos/química , Antineoplásicos/metabolismo , Cisplatino/química , Cisplatino/metabolismo , DNA/metabolismo , Entropia , Pinças ÓpticasRESUMO
We present a new model to describe DNA interactions with large ligands such as proteins, based on a quenched-disorder equation for ligand binding along the double helix and on Manning's description for the local changes of the persistence length at the binding sites. Such a model allows one to extract the physical chemistry of the interactions from pure mechanical measurements, such as those typically performed with DNA-protein complexes in force spectroscopy assays. We have tested the proposed methodology here to investigate the DNA interaction with the protein lysozyme, determining binding parameters such as the equilibrium association constant, the cooperativity degree of the binding reaction, and the fraction of neutralized charges on the phosphate backbone. The model also allows one to get information on the size and positional conformation of the bound proteins.
Assuntos
DNA Viral/química , Muramidase/química , Físico-Química , Ligantes , Fenômenos Mecânicos , Modelos Moleculares , Muramidase/metabolismoRESUMO
In this work we use single molecule force spectroscopy performed with optical tweezers in order to characterize the complexes formed between the anticancer drug Pixantrone (PIX) and the DNA molecule, at two very different ionic strengths. Firstly, the changes of the mechanical properties of the DNA-PIX complexes were studied as a function of the drug concentration in the sample. Then, a quenched-disorder statistical model of ligand binding was used in order to determine the physicochemical (binding) parameters of the DNA-PIX interaction. In particular, we have found that the PIX molecular mechanism of action involves intercalation into the double helix, followed by a significant compaction of the DNA molecule due to partial neutralization of the phosphate backbone. Finally, this scenario of interaction was quantitatively compared to that found for the related drug Mitoxantrone (MTX), which binds to DNA with a considerably higher equilibrium binding constant and promotes a much stronger DNA compaction. The comparison performed between the two drugs can bring clues to the development of new (and more efficient) related compounds.
Assuntos
Antineoplásicos/química , DNA/química , Substâncias Intercalantes/química , Isoquinolinas/química , Ligantes , Pinças Ópticas , Imagem Individual de Molécula/métodosRESUMO
Here, we use single molecule force spectroscopy performed with optical tweezers in order to investigate the interaction between Caffeine and the DNA molecule for various different concentrations of the alkaloid and under two distinct ionic strengths of the surrounding buffer. We were able to determine the mechanical changes induced on the double-helix structure due to Caffeine binding, the binding mode and the binding parameters of the interaction. The results obtained show that Caffeine binds to DNA by outside the double-helix with a higher affinity at lower ionic strengths. On the other hand, a considerable cooperativity was found only for sufficient high ionic strengths, suggesting that Caffeine may binding forming dimers and/or trimers along the double-helix under this condition. Finally, it was also shown that Caffeine stabilizes the DNA double-helix upon binding, preventing force-induced DNA melting.
Assuntos
Cafeína/farmacologia , DNA/química , Conformação de Ácido Nucleico/efeitos dos fármacos , Algoritmos , Cafeína/química , Modelos Teóricos , Estrutura Molecular , Desnaturação de Ácido Nucleico/efeitos dos fármacos , Pinças Ópticas , Análise EspectralRESUMO
Here we report a new study performed at single molecule level on the interaction of the antineoplastic drug Carboplatin and the DNA molecule - the main target of the drug inside cells in cancer chemotherapies. By using optical tweezers, we measure how the mechanical properties of the DNA-Carboplatin complexes changes as a function of the drug concentration in the sample, for two different ionic strengths ([Na]â¯=â¯150â¯mM and [Na]â¯=â¯1â¯mM). From these measurements, the binding mechanism and the physicochemical (binding) parameters of the interaction were inferred and directly compared to those obtained for the precursor drug Cisplatin under equivalent conditions. As the main conclusion, we show that Carboplatin binds preferentially forming covalent monoadducts in contrast to Cisplatin, which is hydrolyzed easier and presents a higher efficiency in forming covalent diadducts along the double-helix. In addition, we explicitly show that Carboplatin is much less sensitive to ionic strength changes when compared to Cisplatin. These findings provide new insights on the interactions of platinum-based compounds with the DNA molecule, being important to improve the current treatments and in the development of new antineoplastic agents.
Assuntos
Carboplatina/metabolismo , Cisplatino/metabolismo , DNA/metabolismo , Carboplatina/química , Carboplatina/uso terapêutico , Cisplatino/química , Cisplatino/uso terapêutico , DNA/química , Humanos , Cinética , Neoplasias/tratamento farmacológico , Conformação de Ácido Nucleico , Desnaturação de Ácido Nucleico , Concentração OsmolarRESUMO
We have performed a biophysical characterization, at single molecule level, of the interaction between the DNA molecule and the biogenic polyamine putrescine. By using force spectroscopy, we were able to monitor the complexes formation as putrescine is added to the sample, determining the mechanical properties of such complexes and the physicochemical (binding) parameters of the interaction for three different ionic strengths. In particular, it was shown that the behavior of the equilibrium binding constant as a function of the counterion concentration deviates from the prediction of the Record-Lohman model. The measured constants were (1.3 ± 0.2)â¯×â¯105â¯M-â¯1 for [Na]â¯=â¯150â¯mM, (2.1 ± 0.2)â¯×â¯105â¯M-â¯1 for [Na]â¯=â¯10â¯mM, and (2.2 ± 0.3)â¯×â¯105â¯M-â¯1 for [Na]â¯=â¯1â¯mM. The cooperativity degree of the binding reaction, on the other hand, increases with the ionic strength. From these analysis, the DNA-putrescine binding mechanisms are inferred, and a comparison with results reported for ordinary bivalent ions like magnesium is performed. Such study provides new insights on the general behavior of the DNA interactions with biogenic polyamines.
Assuntos
Poliaminas Biogênicas/química , Proteínas de Ligação a DNA/química , DNA/química , Nanotecnologia , Sítios de Ligação , Fenômenos Biofísicos , Modelos Moleculares , Conformação de Ácido Nucleico , Concentração Osmolar , Espermidina/química , Espermina/químicaRESUMO
We report a high cooperative transition from the semi-flexible to the flexible regime of polymer elasticity during the interaction of the DNA molecule with the chemotherapeutic drug Mitoxantrone (MTX). By using single molecule force spectroscopy, we show that the force-extension curves of the DNA-MTX complexes deviate from the typical worm-like chain behavior as the MTX concentration in the sample increases, becoming straight lines for sufficiently high drug concentrations. The behavior of the radius of gyration of the complexes as a function of the bound MTX concentration was used to quantitatively investigate the cooperativity of the condensation process. The present methodology can be promptly applied to other ligands that condense the DNA molecule upon binding, opening new possibilities in the investigation of this type of process and, more generally, in the investigation of phase transitions in polymer physics.
Assuntos
DNA/química , Mitoxantrona/químicaRESUMO
This study aimed to assess anthropogenic impact of surrounding population in the Private Reserve of Natural Heritage at Pantanal, the world's largest freshwater wetland ecosystem located in the centre of South America. Viral aetiological agents of acute gastroenteritis as rotavirus A (RVA), noroviruses, human adenoviruses, klassevirus and of hepatitis, as hepatitis A virus, were investigated in different aquatic matrices. Annual collection campaigns were carried out from 2009 to 2012, alternating dry and rainy seasons. Viral particles present in the samples were concentrated by the adsorption-elution method, with negatively charged membranes, and detected by qualitative and quantitative PCR. From a total of 43 samples at least one virus was detected in 65% (28) of them. Viruses were detected in all matrices with concentrations ranging from 2 × 102 to 8·3 × 104 genome copies per litre. A significant higher RVA frequency was observed in the dry season. Our data revealing dissemination of human enteric viruses in water matrices both inside and outside the reserve could be useful to trace faecal contamination in the environment and to minimize the risk of infection by exposure of susceptible individuals. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is part of a collaborative project designed to investigate the environmental and health conditions of the Private Reserve of Natural Heritage at Pantanal, the largest seasonally flooded wetland in the world. The project aimed to promote health and quality of human and wildlife extending technical-scientific knowledge about pathogens present in the region. By assessing the occurrence of human enteric viruses in different water matrices we demonstrated the anthropogenic impact of surrounding population and pointed out the potential risk of infection by exposure of susceptible individuals.
Assuntos
Adenoviridae/isolamento & purificação , Enterovirus/isolamento & purificação , Gastroenterite/virologia , Vírus da Hepatite A/isolamento & purificação , Norovirus/isolamento & purificação , Parques Recreativos , Rotavirus/isolamento & purificação , Doenças Transmitidas pela Água/virologia , Adenoviridae/genética , Antígenos Virais , Brasil/epidemiologia , Ecossistema , Enterovirus/genética , Fezes/virologia , Água Doce/virologia , Gastroenterite/epidemiologia , Vírus da Hepatite A/genética , Humanos , Norovirus/genética , Chuva/virologia , Reação em Cadeia da Polimerase em Tempo Real , Rotavirus/genética , Estações do Ano , Microbiologia da Água , Doenças Transmitidas pela Água/epidemiologiaRESUMO
Maturity-Onset Diabetes of the Young (MODY) type 4 or PDX1 -MODY is a rare form of monogenic diabetes caused by heterozygous variants in PDX1 . Pancreatic developmental anomalies related to PDX1 are reported only in neonatal diabetes cases. Here, we describe dorsal pancreatic agenesis in 2 patients with PDX1 -MODY. The proband presented with diabetes since 14 years of age and maintained regular glycemic control with low doses of basal insulin and detectable C-peptide levels after 38 years with diabetes. A diagnosis of MODY was suspected. Targeted next-generation sequencing identified a heterozygous variant in PDX1 : c.188delC/p.Pro63Argfs*60. Computed tomography revealed caudal pancreatic agenesis. Low fecal elastase indicated exocrine insufficiency. His son had impaired glucose tolerance, presented similar pancreatic agenesis, and harbored the same allelic variant. The unusual presentation in this Brazilian family enabled expansion upon a rare disease phenotype, demonstrating the possibility of detecting pancreatic malformation even in cases of PDX1 -related diabetes diagnosed after the first year of life. This finding can improve the management of MODY4 patients, leading to precocious investigation of pancreatic dysgenesis and exocrine dysfunction.
Assuntos
Anormalidades Congênitas/genética , Diabetes Mellitus Tipo 2/genética , Proteínas de Homeodomínio/genética , Pâncreas/anormalidades , Doenças Raras/genética , Transativadores/genética , Brasil , Peptídeo C/genética , Pré-Escolar , Anormalidades Congênitas/diagnóstico , Anormalidades Congênitas/fisiopatologia , Diabetes Mellitus Tipo 2/diagnóstico , Diabetes Mellitus Tipo 2/fisiopatologia , Intolerância à Glucose/genética , Intolerância à Glucose/fisiopatologia , Heterozigoto , Humanos , Masculino , Pessoa de Meia-Idade , Mutação , Pâncreas/fisiopatologia , Elastase Pancreática/genética , Fenótipo , Doenças Raras/diagnóstico , Doenças Raras/fisiopatologiaRESUMO
In this work, we have studied the interaction between the anticancer drug doxorubicin (doxo) and condensed DNA, using optical tweezers. To perform this task, we use the protein bovine serum albumin (BSA) in the working buffer to mimic two key conditions present in the real intracellular environment: the condensed state of the DNA and the abundant presence of charged macromolecules in the surrounding medium. In particular, we have found that, when doxo is previously intercalated in disperse DNA, the drug hinders the DNA condensation process upon the addition of BSA in the buffer. On the other hand, when bare DNA is firstly condensed by BSA, doxo is capable to intercalate and to unfold the DNA condensates at relatively high concentrations. In addition, a specific interaction between BSA and doxo was verified, which significantly changes the chemical equilibrium of the DNA-doxo interaction. Finally, the presence of BSA in the buffer stabilizes the double-helix structure of the DNA-doxo complexes, preventing partial DNA denaturation induced by the stretching forces.