Imidazolium-based ionic liquids binding to DNA: Mechanical effects and thermodynamics of the interactions.

We performed a robust characterization of the molecular interactions between the DNA molecule and two imidazolium-based ionic liquids (ILs): 1-Butyl-3-methylimidazolium chloride ([bmim]Cl) and 1-Octyl-3-methylimidazolium chloride ([omim]Cl), using single molecule approaches (optical and magnetic tweezers) and bulk techniques (isothermal titration calorimetry and conductivity measurements). Optical and magnetic tweezers allowed us to obtain the changes on the mechanical properties of the DNA complexes formed with both ILs, as well as the relevant physicochemical (binding) parameters of the interaction. Despite the weak binding measured between DNA and the two ILs, we identify a transition on the regime of polymer elasticity of the complexes formed, which results in a relevant DNA compaction for high IL concentrations. In addition, isothermal titration calorimetry and conductivity complemented the single molecule investigation, giving a complete thermodynamic characterization of the interactions and allowing the identification of the most relevant driving forces at various different concentration ranges of the ILs. Based on the results obtained with all the employed techniques, we propose a model for the binding schemes involving DNA and both [bmim]Cl and [omim]Cl.

The predatory mite Neoseiulus californicus (Acari: Phytoseiidae) does not respond for volatiles of maize infested by Tetranychus urticae (Acari: Tetranychidae) / O ácaro predador Neoseiulus californicus (Acari: Phytoseiidae) não responde aos voláteis de milho infestado por Tetranychus urticae (Acari: Tetranychidae)

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.

The predatory mite Neoseiulus californicus (Acari: Phytoseiidae) does not respond for volatiles of maize infested by Tetranychus urticae (Acari: Tetranychidae)

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.

The predatory mite Neoseiulus californicus (Acari: Phytoseiidae) does not respond for volatiles of maize infested by Tetranychus urticae (Acari: Tetranychidae) / O ácaro predador Neoseiulus californicus (Acari: Phytoseiidae) não responde aos voláteis de milho infestado por Tetranychus urticae (Acari: Tetranychidae)

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.

Caffeine modulates the intercalation of drugs on DNA: A study at the single molecule level.

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.

Olfactory response of Trichogramma pretiosum (Hymenoptera: Trichogrammatidae) to volatiles induced by transgenic maize.

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.

The predatory mite Neoseiulus californicus (Acari: Phytoseiidae) does not respond for volatiles of maize infested by Tetranychus urticae (Acari: Tetranychidae).

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.

Bessel beam optical tweezers for manipulating superparamagnetic beads.

We propose a Bessel beam optical tweezers setup that can stably trap superparamagnetic beads. The trap stiffness measured is practically independent of the radius of the Bessel beam and of the bead height (distance from the coverlip of the sample chamber), indicating that the beads can be trapped with high accuracy within a wide range of such parameters. On the other hand, the trap stiffness exhibits the expected linear increase with the laser power, despite the non-negligible absorption coefficient of the superparamagnetic beads. A geometrical optics model that considers spherical aberration and light absorption by the beads was used to predict the optical forces and trap stiffness, showing excellent agreement with the experimental data. We believe the results presented here advance the field of optical trapping manipulation of absorbing magnetic particles, and future applications will involve, for example, the design of new hybrid optomagnetic tweezers.

Hydroxychloroquine Exhibits a Strong Complex Interaction with DNA: Unraveling the Mechanism of Action.

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.

Dodecyltrimethylammonium bromide surfactant effects on DNA: Unraveling the competition between electrostatic and hydrophobic interactions.

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.

On the use of Europium (Eu) for designing new metal-based anticancer drugs.

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 ∼ 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.

Transplatin ineffectiveness against cancer from a molecular perspective: A single-molecule force-spectroscopy study.

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.

Silicon microparticles as handles for optical tweezers experiments.

We study the behavior of silicon microparticles in a 1064 nm Gaussian-beam optical tweezers, showing that this semiconductor can be used to perform different types of optical manipulation experiments. Depending on the focal position and the laser power used, the particles can present an oscillatory dynamics in the tweezers or can be stably 3D-trapped with a trap stiffness that allows the application of femtoNewton forces with accuracy. A new, to the best of our knowledge, interpretation based on the photoexcitation of electrons in the valence band is proposed to explain the oscillations, and the quantities associated with such dynamics (e.g., amplitude, period, etc.) were characterized as a function of relevant parameters to optical tweezers setups.

Model for DNA Interactions with Proteins and Other Large Ligands: Extracting Physical Chemistry from Pure Mechanical Measurements.

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.

Pixantrone anticancer drug as a DNA ligand: Depicting the mechanism of action at single molecule level.

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.

Effects of caffeine on the structure and conformation of DNA: A force spectroscopy study.

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.

Vascular angiotensin AT1 receptor neuromodulation in fetal programming of hypertension.

Fetal stress increases the susceptibility to cardiovascular diseases in adult age, including hypertension, a process known as fetal programming of hypertension (FPH). This study intends to investigate the interplay between vascular sympathetic nervous system (SNS) and RAS, namely the neuromodulatory role exerted by Angiotensin II (Ang II) receptor-1 (AT1) in FPH, and respective contribution for hypertension. METHODS: 6-month old Sprague-Dawley offspring from mothers fed ad-libitum (CONTROL) or with 50% intake during the second half of gestation (maternal undernutrition, MUN) were used. Sympathetic neurotransmission was studied in mesenteric/tail arteries and mesenteric veins by electrically-evoked [3H]-noradrenaline release experiments using RAS drugs. AT1 receptors in sympathetic nerves of mesenteric arteries were investigated by immunohistochemistry and Laser Scanning Confocal Microscopy. RESULTS: Ang II facilitated noradrenaline release in the vessels studied from MUN and CONTROL rats. Losartan induced a tonic facilitation only in MUN vessels. Sympathetic innervation was larger in MUN versus CONTROL vessels. AT1 receptors on sympathetic nerves were present in higher amounts in MUN versus CONTROL vessels. CONCLUSIONS: Findings support that FPH is associated with a vascular hyper-sympathetic activation, involving a tonic facilitation of prejunctional AT1 receptors by endogenous Ang II, which can justify, at least in part, the development of hypertension.

Carboplatin as an alternative to Cisplatin in chemotherapies: New insights at single molecule level.

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] = 150 mM and [Na] = 1 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.

Biophysical characterization of the DNA interaction with the biogenic polyamine putrescine: A single molecule study.

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) × 105 M- 1 for [Na] = 150 mM, (2.1 ± 0.2) × 105 M- 1 for [Na] = 10 mM, and (2.2 ± 0.3) × 105 M- 1 for [Na] = 1 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.

A cooperative transition from the semi-flexible to the flexible regime of polymer elasticity: Mitoxantrone-induced DNA condensation.

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.