Synthesis, biophysical characterization and anti-HIV activity of d(TG3AG) Quadruplexes bearing hydrophobic tails at the 5'-end

Novel conjugated G-quadruplex-forming d(TG3AG) oligonucleotides, linked to hydrophobic groups through phosphodiester bonds at 50-end, have been synthesized as potential anti-HIV aptamers, via a fully automated, online phosphoramidite-based solid-phase strategy. Conjugated quadruplexes showed pronounced anti-HIV activity with some preference for HIV-1, with inhibitory activity invariably in the low micromolar range. The CD and DSC monitored thermal denaturation studies on the resulting quadruplexes, indicated the insertion of lipophilic residue at the 50-end, conferring always improved stability to the quadruplex complex (20 < DTm < 40 C). The data suggest no direct functional relationship between the thermal stability and anti-HIV activity of the folded conjugated G-quartets. It would appear that the nature of the residue at 50 end of the d(TG3AG) quadruplexes plays an important role in the thermodynamic stabilization but a minor influence on the anti-HIV activity. Moreover, a detailed CD and DSC analyses indicate a monophasic behaviour for sequences I and V, while for ODNs (II–IV) clearly show that these quadruplex structures deviate from simple two-state melting, supporting the hypothesis that intermediate states along the dissociation pathway may exis

Retrospective 8-year study on the antibiotic resistance of uropathogens in children hospitalised for urinary tract infection in the emilia-romagna region, italy

The development and spread of antibiotic resistance is an increasingly important global public health problem, even in paediatric urinary tract infection (UTI). In light of the variability in the data, it is necessary to conduct surveillance studies to determine the prevalence of antibiotic resistance in specific geographical areas to optimize therapeutic management. In this observational, retrospective, multicentre study, the medical records of 1801 paediatric patients who were hospital-ised for UTI between January 1st, 2012, and June 30th, 2020, in Emilia-Romagna, Italy, were analysed. Escherichia coli was the most frequently detected pathogen (75.6%), followed by Klebsiella pneumoniae (6.9%) and Pseudomonas aeruginosa (2.5%). Overall, 840 cases (46.7%) were due to anti-microbial-resistant uropathogens: 83 (4.7%) extended spectrum beta-lactamase (ESBL)-producing, 119 (6.7%) multidrug resistant (MDR) and 4 (0.2%) extensively drug resistant (XDR) bacteria. Empirical antibiotic therapy failed in 172 cases (9.6%). Having ESBL or MDR/XDR uropathogens, a history of recurrent UTI, antibiotic therapy in the preceding 30 days, and empirical treatment with amoxicillin or amoxicillin/clavulanate were significantly associated with treatment failure, whereas first-line therapy with third-generation cephalosporins was associated with protection against negative outcomes. In conclusion, the increase in the resistance of uropathogens to commonly used antibiotics requires continuous monitoring, and recommendations for antibiotic choice need updating. In our epidemiological context, amoxicillin/clavulanate no longer seems to be the appropriate first-line therapy for children hospitalised for UTI, whereas third-generation cephalosporins continue to be useful. To further limit the emergence of resistance, every effort to reduce and ration-alise antibiotic consumption must be implemented

Clinical Outcome of Discordant Empirical Therapy and Risk Factors Associated to Treatment Failure in Children Hospitalized for Urinary Tract Infections

With the spread of antibiotic resistance in pediatric urinary tract infections (UTIs), more patients are likely to be started empirically on antibiotics to which pathogens are later found to be resistant (discordant therapy). However, in-vivo effectiveness may be different from in-vitro susceptibility. Aims of this study were to describe clinical outcomes of discordant empirical treatments in pediatric UTIs and to investigate risk factors associated to treatment failure. This observational, retrospective study was conducted on children hospitalized for febrile UTIs with positive urine culture and started on discordant empirical therapy. Failure rates of discordant treatments and associated risk factors were investigated. A total of 142/1600 (8.9%) patients were treated with inadequate empirical antibiotics. Clinical failure was observed in 67/142 (47.2%) patients, with no fatal events. Higher failure rates were observed for combinations of penicillin and beta-lactamase inhibitors (57.1%). Significant risk factors for failure of discordant treatment were history of recurrent UTIs (95% CI: 1.13–9.98, OR: 3.23, p &lt; 0.05), recent use of antibiotics (95% CI: 1.46–21.82, OR: 5.02, p &lt; 0.01), infections caused by Pseudomonas aeruginosa (95% CI: 1.85–62.10, OR: 7.30, p &lt; 0.05), and empirical treatment with combinations of penicillin and beta-lactamase inhibitors (95% CI: 0.94–4.03, OR: 1.94, p = 0.05). This study showed that discordant empirical treatments may still be effective in more than half of pediatric UTIs. Clinical effectiveness varies between different discordant antibiotics in pediatric UTIs, and patients presenting risk factors for treatment failure may need a differentiated empirical approach

Functional diversity of nanohaloarchaea within xylan-degrading consortia

DATA AVAILABILITY STATEMENT : The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found in the article/Supplementary material.Extremely halophilic representatives of the phylum Candidatus Nanohaloarchaeota (members of the DPANN superphyla) are obligately associated with extremely halophilic archaea of the phylum Halobacteriota (according to the GTDB taxonomy). Using culture-independent molecular techniques, their presence in various hypersaline ecosystems around the world has been confirmed over the past decade. However, the vast majority of nanohaloarchaea remain uncultivated, and thus their metabolic capabilities and ecophysiology are currently poorly understood. Using the (meta)genomic, transcriptomic, and DNA methylome platforms, the metabolism and functional prediction of the ecophysiology of two novel extremely halophilic symbiotic nanohaloarchaea (Ca. Nanohalococcus occultus and Ca. Nanohalovita haloferacivicina) stably cultivated in the laboratory as members of a xylose-degrading binary culture with a haloarchaeal host, Haloferax lucentense, was determined. Like all known DPANN superphylum nanoorganisms, these new sugar-fermenting nanohaloarchaea lack many fundamental biosynthetic repertoires, making them exclusively dependent on their respective host for survival. In addition, given the cultivability of the new nanohaloarchaea, we managed to discover many unique features in these new organisms that have never been observed in nano-sized archaea both within the phylum Ca. Nanohaloarchaeota and the entire superphylum DPANN. This includes the analysis of the expression of organism-specific non-coding regulatory (nc)RNAs (with an elucidation of their 2D-secondary structures) as well as profiling of DNA methylation. While some ncRNA molecules have been predicted with high confidence as RNAs of an archaeal signal recognition particle involved in delaying protein translation, others resemble the structure of ribosome-associated ncRNAs, although none belong to any known family. Moreover, the new nanohaloarchaea have very complex cellular defense mechanisms. In addition to the defense mechanism provided by the type II restriction-modification system, consisting of Dcm-like DNA methyltransferase and Mrr restriction endonuclease, Ca. Nanohalococcus encodes an active type I-D CRISPR/Cas system, containing 77 spacers divided into two loci. Despite their diminutive genomes and as part of their host interaction mechanism, the genomes of new nanohaloarchaea do encode giant surface proteins, and one of them (9,409 amino acids long) is the largest protein of any sequenced nanohaloarchaea and the largest protein ever discovered in cultivated archaea.The FUTURENZYMES Project funded by the European Union's Horizon 2020 Research Program; Agence Nationale de la Recherche grant; the project “Emergence of virulence and antibiotic-resistance vectors in coastal and deep sea marine environments and analysis of the mechanisms and conditions underlying their spread and evolution” funded by the MIUR Progetti di Ricerca di Rilevante Interesse Nazionale (PRIN) Bando 2017; nd by the project “Innovazione tecnologica per la tutela e valorizzazione degli ecosistemi marini (ITEM)” funded by MIUR CTN02_00059_9948371.frontiersin.orghttp://www.frontiersin.org/Microbiologyam2024BiochemistryGeneticsMicrobiology and Plant PathologySDG-15:Life on lan

Perioperative Care in Cardiac Surgery: A Joint Consensus Statement by the Enhanced Recovery After Surgery (ERAS) Cardiac Society, ERAS International Society, and The Society of Thoracic Surgeons (STS).

Enhanced Recovery After Surgery (ERAS) programs have been shown to lessen surgical insult, promote recovery, and improve postoperative clinical outcomes across a number of specialty operations. A core tenet of ERAS involves the provision of protocolized evidence-based perioperative interventions. Given both the growing enthusiasm for applying ERAS principles to cardiac surgery and the broad scope of relevant interventions, an international, multidisciplinary expert panel was assembled to derive a list of potential program elements, review the literature, and provide a statement regarding clinical practice for each topic area. This article summarizes those consensus statements and their accompanying evidence. These results provide the foundation for best practice for the management of the adult patient undergoing cardiac surgery

Nanohaloarchaea as beneficiaries of xylan degradation by haloarchaea

DATA AVAILABILITY STATEMENT : All statements regarding data availability, finding resources and conflict of interest disclosure have been provided.DATA DEPOSITION : All (meta)genomic and transcriptomic information is available under GenBank BioProject ID PRJNA865582; BioSamples SAMN30630960, SAMN30630938, SAMN30630946, SAMN30631035, SAMN30630966, SAMN30631033, SAMN30120999, SAMN30121000, SAMN30121001, SAMN30121002, SAMN30121003 and SAMN30121004; genome accession no. CP104322 for Halorhabdus sp. SVX81, genome accession no. CP104395 for Ca. Nanohalococcus occultus SVXNc, genome accession nos. CP104741 (chromosome), CP104742 (plasmid 1), CP104743 (plasmid 2) and CP104744 (plasmid 3) for Haloferax lucertense SVX82; genome accession no. CP107254 for Halorhabdus sp. BNX81, genome accession no. CP107255 for Ca. Nanohalovita haloferacivicina BNXNv, genome accession nos. CP106966 (chromosome), CP106967 (plasmid 1), CP106968 (plasmid 2), CP106969 (plasmid 3) and CP106970 (plasmid 4) for Haloferax lucertense BNX82. Transcriptomic raw data are available under accession SRR21676140-SRR216761401. The mass spectrometry proteomics data have been deposited in the ProteomeXchange Consortium via the PRIDE partner repository with the data set identifier PXD036877.Climate change, desertification, salinisation of soils and the changing hydrology of the Earth are creating or modifying microbial habitats at all scales including the oceans, saline groundwaters and brine lakes. In environments that are saline or hypersaline, the biodegradation of recalcitrant plant and animal polysaccharides can be inhibited by salt-induced microbial stress and/or by limitation of the metabolic capabilities of halophilic microbes. We recently demonstrated that the chitinolytic haloarchaeon Halomicrobium can serve as the host for an ectosymbiont, nanohaloarchaeon ‘Candidatus Nanohalobium constans’. Here, we consider whether nanohaloarchaea can benefit from the haloarchaea-mediated degradation of xylan, a major hemicellulose component of wood. Using samples of natural evaporitic brines and anthropogenic solar salterns, we describe genome-inferred trophic relations in two extremely halophilic xylan-degrading three-member consortia. We succeeded in genome assembly and closure for all members of both xylan-degrading cultures and elucidated the respective food chains within these consortia. We provide evidence that ectosymbiontic nanohaloarchaea is an active ecophysiological component of extremely halophilic xylan-degrading communities (although by proxy) in hypersaline environments. In each consortium, nanohaloarchaea occur as ectosymbionts of Haloferax, which in turn act as scavenger of oligosaccharides produced by xylan-hydrolysing Halorhabdus. We further obtained and characterised the nanohaloarchaea–host associations using microscopy, multi-omics and cultivation approaches. The current study also doubled culturable nanohaloarchaeal symbionts and demonstrated that these enigmatic nano-sized archaea can be readily isolated in binary co-cultures using an appropriate enrichment strategy. We discuss the implications of xylan degradation by halophiles in biotechnology and for the United Nation's Sustainable Development Goals.Agence Nationale de la Recherche; Centre for Environmental Biotechnology Project, partly funded by the European Regional Development Fund via the Welsh Assembly Government European Union; H2020 Food; Ministerio de Ciencia e Innovación, Agencia Estatal de Investigación (AEI);; SYAM-Gravitation Program of the Dutch Ministry of Education, Culture and Science.https://sfamjournals.onlinelibrary.wiley.com/journal/17517915am2024BiochemistryGeneticsMicrobiology and Plant PathologySDG-03:Good heatlh and well-beingSDG-08:Decent work and economic growthSDG-09: Industry, innovation and infrastructureSDG-11:Sustainable cities and communitiesSDG-12:Responsible consumption and productionSDG-13:Climate actionSDG-14:Life below waterSDG-15:Life on lan

Developmental Issues in School-Based Aggression Prevention from a Social-Cognitive Perspective

Contemporary research on the development and prevention of aggressive behavior in childhood and adolescence emphasizes the importance of social-cognitive factors such as perceptual biases, problem-solving skills, and social-moral beliefs in the maintenance of aggression. Indeed, school-based social-cognitive intervention approaches have been identified as best practices by the Centers for Disease Control and Prevention. However, because child age is an important covariate of both intervention effectiveness and social-cognitive ability, school-based prevention program designers should keep in mind a number of issues identified through developmental research. In this paper, we review the social-cognitive model of aggressive behavior development as applied to prevention programming. We then discuss some of the ways in which the broader developmental research base can inform the design of aggression prevention programs.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/45098/1/10935_2005_Article_5.pd

Lock-In Based Differential Front-End for Raman Spectroscopy Applications

The intrinsic sensitivity limit of Stimulated Raman Spectroscopy (SRS) is given by the shot noise of the optical stimulation. However, it is seldom reached due to the electronic noise of the front-end amplifier and the intensity fluctuations of the laser source. Here, we present and test a low-noise pseudo-differential amplifier, for Raman spectroscopy applications, able to compensate the common-mode fluctuations given by the laser and to reach a sensitivity better than 10 ppm thanks to the lock-in technique