The oxidative damage to the human telomere: effects of 5-hydroxymethyl-2'-deoxyuridine on telomeric G-quadruplex structures

As part of the genome, human telomeric regions can be damaged by the chemically reactive molecules responsible for oxidative DNA damage. Considering that G-quadruplex structures have been proven to occur in human telomere regions, several studies have been devoted to investigating the effect of oxidation products on the properties of these structures. However only investigations concerning the presence in G-quadruplexes of the main oxidation products of deoxyguanosine and deoxyadenosine have appeared in the literature. Here, we investigated the effects of 5-hydroxymethyl-2’-deoxyuridine (5-hmdU), one of the main oxidation products of T, on the physical–chemical properties of the G-quadruplex structures formed by two human telomeric sequences. Collected calorimetric, circular dichroism and electrophoretic data suggest that, in contrast to most of the results on other damage, the replacement of a T with a 5-hmdU results in only negligible effects on structural stability. Reported results and other data from literature suggest a possible protecting effect of the loop residues on the other parts of the G-quadruplexes

Strand directionality affects cation binding and movement within tetramolecular G-quadruplexes

Nuclear magnetic resonance study of G-quadruplex structures formed by d(TG3T) and its modified analogs containing a 50-50 or 30-30 inversion of polarity sites, namely d(30TG50-50G2T30), d(30T50- 50G3T30) and d(50TG30-30G2T5’) demonstrates formation of G-quadruplex structures with tetrameric topology and distinct cation-binding preferences. All oligonucleotides are able to form quadruplex structures with two binding sites, although the modified oligonucleotides also form, in variable amounts, quadruplex structures with only one bound cation. The inter-quartet cavities at the inversion of polarity sites bind ammonium ions less tightly than a naturally occurring 50-30 backbone. Exchange of 15NH+ 4 ions between G-quadruplex and bulk solution is faster at the 30-end in comparison to the 50-end. In addition to strand directionality, cation movement is influenced by formation of an all-syn G-quartet. Formation of such quartet has been observed also for the parent d(TG3T) that besides the canonical quadruplex with only all-anti G-quartets, forms a tetramolecular parallel quadruplex containing one all-syn G-quartet, never observed before in unmodified quadruplex structures

Connectivity alterations underlying the breakdown of pseudoneglect: New insights from healthy and pathological aging

A right-hemisphere dominance for visuospatial attention has been invoked as the most prominent neural feature of pseudoneglect (i.e., the leftward visuospatial bias exhibited in neurologically healthy individuals) but the neurophysiological underpinnings of such advantage are still controversial. Previous studies investigating visuospatial bias in multiple-objects visual enumeration reported that pseudoneglect is maintained in healthy elderly and amnesic mild cognitive impairment (aMCI), but not in Alzheimer's disease (AD). In this study, we aimed at investigating the neurophysiological correlates sustaining the rearrangements of the visuospatial bias along the progression from normal to pathological aging. To this aim, we recorded EEG activity during an enumeration task and analyzed intra-hemispheric fronto-parietal and inter-hemispheric effective connectivity adopting indexes from graph theory in patients with mild AD, patients with aMCI, and healthy elderly controls (HC). Results revealed that HC showed the leftward bias and stronger fronto-parietal effective connectivity in the right as compared to the left hemisphere. A breakdown of pseudoneglect in patients with AD was associated with both the loss of the fronto-parietal asymmetry and the reduction of inter-hemispheric parietal interactions. In aMCI, initial alterations of the attentional bias were associated with a reduction of parietal inter-hemispheric communication, but not with modulations of the right fronto-parietal connectivity advantage, which remained intact. These data provide support to the involvement of fronto-parietal and inter-parietal pathways in the leftward spatial bias, extending these notions to the complex neurophysiological alterations characterizing pathological aging

Improved performances of catalytic G-quadruplexes (G4-DNAzymes) via the chemical modifications of the DNA backbone to provide Gquadruplexes with double 3′-external G-quartets

Here we report on the design of a new catalytic G-quadruplex-DNA system (G4-DNAzyme) based on the modification of the DNA scaffold to provide the DNA pre-catalyst with two identical 3′-ends, known to bemore catalytically proficient than the 5′-ends. To this end, we introduced a 5′-5′ inversion of polarity site in the middle of the G4-forming sequences AG4A andAG6A to obtain d(3′AGG5′-5′GGA3′) (orAG2-G2A) and d(3′AGGG5′-5′GGGA3′) (or AG3-G3A) that fold into stable G4 whose tetramolecular nature was confirmed via nuclear magnetic resonance (NMR) and circular dichroism(CD) investigations. Both AG2-G2AandAG3-G3A display two identical external G-quartets (3′-ends) known to interact with the cofactor hemin with a high efficiency, making the resulting complex competent to performhemoprotein-like catalysis (G4-DNAzyme). A systematic comparison of the performances of modified and unmodified G4s lends credence to the relevance of the modification exploited here (5′-5′ inversion of polarity site), which represents a new chemical opportunity to improve the overall activity of catalytic G4s

Exploring the binding of d(GGGT)4 to the HIV-1 integrase: An approach to investigate G-quadruplex aptamer/target protein interactions.

The aptamer d(GGGT)4 (T30923 or T30695) forms a 5'-5' dimer of two stacked parallel G-quadruplexes, each characterized by three G-tetrads and three single-thymidine reversed-chain loops. This aptamer has been reported to exhibit anti-HIV activity by targeting the HIV integrase, a viral enzyme responsible for the integration of viral DNA into the host-cell genome. However, information concerning the aptamer/ target interaction is still rather limited. In this communication we report microscale thermophoresis investigations on the interaction between the HIV-1 integrase and d(GGGT)4 aptamer analogues containing abasic sites singly replacing thymidines in the original sequence. This approach has allowed the identification of which part of the aptamer G-quadruplex structure is mainly involved in the interaction with the protei

Psychologists in italian hospital settings: an exploratory analysis of hospital physicians: representations and demands of psychological Intervention

Aunque la Organización Mundial de la Salud ha evidenciado la importancia de adoptar modelos integrados de asistencia basados en el enfoque bio-psico-social, en los servicios de salud, como los hospitales, existe todavía mucha resistencia para su implementación. Se presenta un estudio exploratorio cualitativo dirigido a comprender las representaciones y las demandas de los médicos hospitalarios con respecto a la intervención psicológica en los contextos hospitalarios italianos. Conforme a este objetivo se desarrolló un guión de entrevista semi-estructurada y se entrevistaron 15 médicos italianos divididos por género, edad y especialidad médica. Fueron investigados las representaciones de la enfermedad, el acercamiento a los pacientes y los modelos de intervención, la experiencia de colaboración con psicólogos, las representaciones de dónde, cuándo y cómo los psicólogos podrían ser útiles en el contexto hos pitalario, las visiones generales del hospital y de su futuro, los puntos de fuerza y de debilidad de su trabajo, las estrategias utilizadas para superar las dificultades diarias y las estrategias propuestas pa ra mejorar el sistema hospitalario. El análisis de contenido de las entrevistas ha revelado que la intervención psicológica está representada predominantemente como una intervención a nivel individual dirigida a contener el distrés emocional en relación con la enfermedad. Además se ha evidenciado la falta de consideración de la competencia psicológica como útil para promocionar el funcionamiento organizativo de los servicios hospitalarios. Estos resultados pueden apoyar la investigación futura en esta área y son utilizados pa - ra discutir las oportunidades de un diálogo constructivo entre ciencias médica y psicológica en los contextos hospitalarios.ABSTRACT: During the years, the World Health Organiza - tion has been emphasising the importance of promoting integrated models of care consistent with a bio-psycho-social approach to health-illness. Nonetheless, healthcare services still show much resistance to the implementation of such models and the use of multidisciplinary teams. As a con - sequence, and despite the efforts made over the years by psychologists, the role and specificity of psychological competence in healthcare settings continues to remain unclear. This is particularly true as regards hospital settings. Based on these considerations, this article presents an exploratory qualitative study aimed at understanding hospital physicians’ representations of, and demands for, psychological intervention in Italian hospital set - tings. The main argument on which the study was based is that the integration of psychological competence into hospital settings very much depends on other professionals’ representation of the psychological profession, as well as on the emotional symbolization of their work context and professional practice. A key role is played by physicians, whose representations of the psychol - ogical intervention in hospital settings have been under studied at the international level, and almost not studied with regard to the Italian context. The purpose of this study was to respond to this gap in the literature. In this regard, a semi-structured interview script was developed and used to inter - view 15 Italian hospital physicians varying in gender, age, and medical speciality. Participants were selected by convenience and interviews were carried out individually in the hospital settings. Representations of disease; approach to patients and models of intervention; experiences of collab - oration with psychologists; representations of where, when and how psychologists could be use - ful in the hospital context; overall visions of one’s hospital and its future; strengths and weaknesses of one’s own work; strategies used to overcome every day difficulties and strategies advocated to improve the hospital were focus of analysis. The interviews, administered by two researchers, were audio-recorded and transcribed. Then a content analysis was developed by three analysts that cod - ed the units of the text (sequences) according to the ca tegories and subcategories identified. The cate - gories were: (a) perspective on medical inter vention; (b) problems and weak nesses; (c) strengths; (d) current and future strategies; (e) psychosocial component of disease, and (f) relation ship between Medicine and Psychology. Transcripts were ana - lyzed independently by the analysts, and the interrater agreement was 83%. Results revealed a widespread view that psychol ogical science is of limited relevance in the hospital context. The difficulty in perceiving psychologists as a medical resource appeared to be largely related to phys - icians’ endorsement of a bio-medical ap proach, as well as to their lack of opportunities for formal and/or informal collaboration with psy chologists. Psychologists were thought of primarily as diag - nosticians or therapists, and psychological interven tion was mainly characterized as an individuallevel intervention whose main objective is to support the patient and manage his / her emotional response to the disease (and the response of his / her family). Despite physicians felt the hospital system plagued by problems, especially at the structural and functional levels, they did not envisage psychological competence as useful for fostering hospital organisational functioning. On the contrary, they advocated a top-down change in the hospital system, delegating responsibility for change to hospital management or policy makers. As a consequence, an urgent demand for Psychol - ogy remains unexpressed. Although based on a small sample, the present study provides useful insights which may prompt further research in this area. Furthermore, it provides information about the demand for psychological intervention in the hospital context which can be used to develop strategies to promote multidisciplinary care and integrated mind-body interventions. Results sug gest the necessity to rethink professional roles and to promote a constructive dialogue between medical and psychological science in Italian hospital set - tings.info:eu-repo/semantics/publishedVersio

Monomolecular G-quadruplex structures with inversion of polarity sites: new topologies and potentiality

In this paper, we report investigations, based on circular dichroism, nuclear magnetic resonance spectroscopy and electrophoresis methods, on three oligonucleotide sequences, each containing one 3- 3 and two 5-5 inversion of polarity sites, and four G-runs with a variable number of residues, namely two, three and four (mTG2T, mTG3T andmTG4T with sequence 3-TGnT-5-5-TGnT-3-3-TGnT-5-5-TGnT-3 in which n = 2, 3 and 4, respectively), in comparison with their canonical counterparts (TGnT)4 (n = 2, 3 and 4). Oligonucleotides mTG3T and mTG4 T have been proven to form very stable unprecedented monomolecular parallel G-quadruplex structures, characterized by three side loops containing the inversion of polarity sites. Both G-quadruplexes have shown an all-syn G-tetrad, while the other guanosines adopt anti glycosidic conformations. All oligonucleotides investigated have shown a noteworthy antiproliferative activity against lung cancer cell line Calu 6 and colorectal cancer cell line HCT-116 p53−/−. Interestingly, mTG3T andmTG4T have proven to be mostly resistant to nucleases in a fetal bovine serum assay. The whole of the data suggest the involvement of specific pathways and targets for the biological activity

Exploring New Potential Anticancer Activities of the G-Quadruplexes Formed by [(GTG2T(G3T)3] and Its Derivatives with an Abasic Site Replacing Single Thymidine

In this paper, we report our investigations on five T30175 analogues, prepared by replacing sequence thymidines with abasic sites (S) one at a time, in comparison to their natural counterpart in order to evaluate their antiproliferative potential and the involvement of the residues not belonging to the central core of stacked guanosines in biological activity. The collected NMR (Nuclear Magnetic Resonance), CD (Circular Dichroism), and PAGE (Polyacrylamide Gel Electrophoresis) data strongly suggest that all of them adopt G-quadruplex (G4) structures strictly similar to that of the parent aptamer with the ability to fold into a dimeric structure composed of two identical G-quadruplexes, each characterized by parallel strands, three all-anti-G-tetrads and four one-thymidine loops (one bulge and three propeller loops). Furthermore, their antiproliferative (MTT assay) and anti-motility (wound healing assay) properties against lung and colorectal cancer cells were tested. Although all of the oligodeoxynucleotides (ODNs) investigated here exhibited anti-proliferative activity, the unmodified T30175 aptamer showed the greatest effect on cell growth, suggesting that both its characteristic folding in dimeric form and its presence in the sequence of all thymidines are crucial elements for antiproliferative activity. This straightforward approach is suitable for understanding the critical requirements of the G-quadruplex structures that affect antiproliferative potential and suggests its application as a starting point to facilitate the reasonable development of G-quadruplexes with improved anticancer properties

Structural and Biological Features of G-Quadruplex Aptamers as Promising Inhibitors of the STAT3 Signaling Pathway

: In this paper, we investigate the structural and biological features of G-quadruplex (G4) aptamers as promising antiproliferative compounds affecting the STAT3 signalling pathway. Targeting the STAT3 protein through high-affinity ligands to reduce its levels or activity in cancer has noteworthy therapeutic potential. T40214 (STAT) [(G3C)4] is a G4 aptamer that can influence STAT3 biological outcomes in an efficient manner in several cancer cells. To explore the effects of an extra cytidine in second position and/or of single site-specific replacements of loop residues in generating aptamers that can affect the STAT3 biochemical pathway, a series of STAT and STATB [GCG2(CG3)3C] analogues containing a thymidine residue instead of cytidines was prepared. NMR, CD, UV, and PAGE data suggested that all derivatives adopt dimeric G4 structures like that of unmodified T40214 endowed with higher thermal stability, keeping the resistance in biological environments substantially unchanged, as shown by the nuclease stability assay. The antiproliferative activity of these ODNs was tested on both human prostate (DU145) and breast (MDA-MB-231) cancer cells. All derivatives showed similar antiproliferative activities on both cell lines, revealing a marked inhibition of proliferation, particularly at 72 h at 30 µM. Transcriptomic analysis aimed to evaluate STAT's and STATB's influence on the expression of many genes in MDA-MB-231 cells, suggested their potential involvement in STAT3 pathway modulation, and thus their interference in different biological processes. These data provide new tools to affect an interesting biochemical pathway and to develop novel anticancer and anti-inflammatory drugs

Platelet abnormalities and platelet-to-lymphocyte ratios in canine immunosuppressant-responsive and non-responsive enteropathy: A retrospective study in 41 dogs

Few studies have examined platelet alterations in dogs with chronic enteropathy. Our aim was to investigate platelet count (PLT), mean platelet volume (MPV), and platelet-to-lymphocyte ratio (PLR) in dogs diagnosed with immunosuppressant-responsive enteropathy (IRE). In this retrospective study of 41 dogs, data regarding signalment, canine chronic enteropathy clinical activity index (CCECAI), endoscopic and histopathological scores, PLT, MPV, PLR, total serum protein concentrations, albumin, and iron were collected. Clinical response and relapse were assessed with the evaluation of CCECAI over time. One month after starting therapy, dogs with >25% CCECAI reduction were considered responders. During a three-month CCECAI evaluation as part of a twelve-month follow-up, a CCECAI >3 together with a ≥2 unit increase in responder dogs was considered a relapse. PLT and PLR displayed significant negative correlation with MPV. MPV was positively correlated with total protein and albumin levels and negatively correlated with CCECAI. Three dogs were classified as non-responders, and 14 relapsed within 12 months. No differences were observed in PLT, MPV, or PLR between responding/non-responding and relapsing/non-relapsing groups. PLT, MPV, and PLR correlated with total protein, albumin, and CCECAI, confirming PLT as a potential marker, and suggesting MPV as a new marker of clinical efficacy against canine IRE