Ctf18-RFC and DNA Pol ϵ form a stable leading strand polymerase/clamp loader complex required for normal and perturbed DNA replication

The eukaryotic replisome must faithfully replicate DNA and cope with replication fork blocks and stalling, while simultaneously promoting sister chromatid cohesion. Ctf18-RFC is an alternative PCNA loader that links all these processes together by an unknown mechanism. Here, we use integrative structural biology combined with yeast genetics and biochemistry to highlight the specific functions that Ctf18-RFC plays within the leading strand machinery via an interaction with the catalytic domain of DNA Pol ϵ. We show that a large and unusually flexible interface enables this interaction to occur constitutively throughout the cell cycle and regardless of whether forks are replicating or stalled. We reveal that, by being anchored to the leading strand polymerase, Ctf18-RFC can rapidly signal fork stalling to activate the S phase checkpoint. Moreover, we demonstrate that, independently of checkpoint signaling or chromosome cohesion, Ctf18-RFC functions in parallel to Chl1 and Mrc1 to protect replication forks and cell viability. [Abstract copyright: © The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.

A Mediational Model to Explain the Job Insecurity-Emotional Exhaustion Relationship: Breach of Psychological Contract and Distributive Injustice as Intervening Variables

Empirical evidence considers job insecurity as a stressor that leads to poor job-related well-being. With the present study we intend to increase this knowledge by testing how and through what mechanisms job insecurity may give rise to such consequences. In particular, we examined the mediating processes underlying the relationship between job insecurity and emotional exhaustion, as an indicator of psychological well-being and core element of burnout. A total of 322 blue collar workers in Italy are used to test the hypotheses derived from our framework. The results found support for a model in which the effect of job insecurity on exhaustion was mediated by two variables indicating lack of reciprocity, i.e. breached psychological contract and perceived distributive injustice (partial mediational model). Thus, the effect of job insecurity on burnout could partly be interpreted in terms of inequity perceived from the exchange relationship between employee and organization. The tested model provides a theoretical framework that may lead to new insights on the job insecurity-exhaustion relationship and on the development of burnout in the uncertain organizational context. Results are discussed and theoretical and practical implications are suggested

Role of transoesophageal echocardiography in evaluating the effect of catheter ablation of atrial fibrillation on anatomy and function of the pulmonary veins

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First cases of combined full robotic partial nephrectomy and colorectal resections: Results and new perspectives

Background: Nowadays the robotic platform is widespread in general surgery, urology, and gynecology. Combined surgery may represent an alternative to sequential procedures and it allows the treatment, at the same time, of coexisting lesions; in this perspective, full-robotic multiorgan surgery is starting to gain interest from surgeons worldwide. Methods: Between April and June 2019, two patients presenting with synchronous colorectal and kidney cancers underwent, respectively, full-robotic right colectomy with right partial nephrectomy and anterior rectal resection with left partial nephrectomy. Surgeries were performed by both the general surgery and urology team. Results: No intraoperative complications were registered and the postoperative course was uneventful in both cases. Conclusions: Combined multiple organ surgery with full robotic technique is safe and offers oncological adequate results. A multi-team surgical pre-planning is mandatory to reduce invasiveness and operative time. To the best of our knowledge, these are the first reports of full robotic partial nephrectomy combined with colorectal procedures

New intravenous calcimimetic agents: New options, new problems. an example on how clinical, economical and ethical considerations affect choice of treatment

Background. Dialysis treatment is improving, but several long-term problems remain unsolved, including metabolic bone disease linked to chronic kidney disease (CKD-MBD). The availability of new, efficacious but expensive drugs (intravenous calcimimetic agents) poses ethical problems, especially in the setting of budget limitations. Methods. Reasons of choice, side effects, biochemical trends were discussed in a cohort of 15 patients (13% of the dialysis population) who stared treatment with intravenous calcimimetics in a single center. All patients had previously been treated with oral calcimimetic agents; dialysis efficacy was at target in 14/15; hemodiafiltration was employed in 10/15. Median Charlson Comorbidity Index was 8. The indications were discussed according to the principlist ethics (beneficience, non maleficience, justice and autonomy). Biochemical results were analyzed to support the clinical-ethical choices. Results. In the context of a strict clinical and biochemical surveillance, the lack of side effects ensured “non-maleficence”; efficacy was at least similar to oral calcimimetic agents, but tolerance was better. Autonomy was respected through a shared decision-making model; all patients appreciated the reduction of the drug burden, and most acknowledged better control of their biochemical data. The ethical conflict resides in the balance between the clinical “beneficience, non-maleficience” advantage and “justice” (economic impact of treatment, potentially in attrition with other resources, since the drug is expensive and included in the dialysis bundle). The dilemma is more relevant when a patient’s life expectancy is short (economic impact without clear clinical advantages), or when non-compliance is an issue (unclear advantage if the whole treatment is not correctly taken). Conclusions. In a context of person-centered medicine, autonomy, beneficence and non-maleficence should weight more than economic justice. While ethical discussions are not aimed at finding “the right answer” but asking “the right questions”, this example can raise awareness of the importance of including an ethical analysis in the choice of “economically relevant” drugs

Predição da resistência genética ao carrapato de bovinos Braford e Hereford a partir de um painel denso de marcadores moleculares.

O problema do carrapato bovino; A alternativa da seleção genômica; Resultados experimentais; Procedimentos para aplicação prática; Considerações finais.bitstream/item/77131/1/CT41.pd

Smc5/6: a link between DNA repair and unidirectional replication?

Of the three structural maintenance of chromosome (SMC) complexes, two directly regulate chromosome dynamics. The third, Smc5/6, functions mainly in homologous recombination and in completing DNA replication. The literature suggests that Smc5/6 coordinates DNA repair, in part through post-translational modification of uncharacterized target proteins that can dictate their subcellular localization, and that Smc5/6 also functions to establish DNA-damage-dependent cohesion. A nucleolar-specific Smc5/6 function has been proposed because Smc5/6 yeast mutants display penetrant phenotypes of ribosomal DNA (rDNA) instability. rDNA repeats are replicated unidirectionally. Here, we propose that unidirectional replication, combined with global Smc5/6 functions, can explain the apparent rDNA specificity

Contextual influences on italian university students during the covid-19 lockdown: Emotional responses, coping strategies and resilience

none17Based on an ecological perspective on the COVID-19 lockdown experience, this study describes psychological responses among Italian university students. Our study considers three zones of the country that have differed in the intensity of the COVID-19 pandemic. Specifically, this research explores whether differences in pandemic conditions can account for their divergent psychological outcomes. The participants were 792 university students from seven different Italian universities. Students were asked to express their emotions and describe meaningful events during the lockdown in writing. Based on the grounded theory approach, this study conducted qualitative data analysis using ATLAS.ti 8.0. The core emerged categories are emotions, emotional moods and state of mind, coping strategies, and resilience. The results describing these emergent factors in relation to environmental variables highlight differences in the feeling of anxiety among individuals: anxiety was more self-focused in zones that were more affected by the lockdown, while such anxiety was more related to family and friends in less-affected zones. In addition to identifying the negative repercussions that this emergency has had, this study describes some positive outcomes, such as the elaboration of new personal perspectives that help foster individual growth and allow individuals to gain new awareness of themselves and others. The confinement due to the COVID-19 emergency measures has been a very unique experience for people, and further research is needed to understand the long-term effects of the different coping responses activated by participants during and after the lockdown.mixedLaura Migliorini; Norma De Piccoli; Paola Cardinali; Chiara Rollero; Daniela Marzana; Caterina Arcidiacono; Elisa Guidi; Ciro Esposito; Cinzia Novara; Angela Fedi; Elena Marta; Andrea Guazzini; Patrizia Meringolo; Maria Grazia Monaci; Barbara Agueli; Fortuna Procentese; Immacolata Di NapoliMigliorini, Laura; De Piccoli, Norma; Cardinali, Paola; Rollero, Chiara; Marzana, Daniela; Arcidiacono, Caterina; Guidi, Elisa; Esposito, Ciro; Novara, Cinzia; Fedi, Angela; Marta, Elena; Guazzini, Andrea; Meringolo, Patrizia; Grazia Monaci, Maria; Agueli, Barbara; Procentese, Fortuna; Di Napoli, Immacolat

Effect of resveratrol on mitochondrial function: Implications in parkin-associated familiar Parkinson's disease

Mitochondrial dysfunction and oxidative stress occur in Parkinson's disease (PD), but the molecular mechanisms controlling these events are not completely understood. Peroxisome proliferator-activated receptor-gamma coactivator-1α (PGC-1α) is a transcriptional coactivator known as master regulator of mitochondrial functions and oxidative metabolism. Recent studies, including one from our group, have highlighted altered PGC-1α activity and transcriptional deregulation of its target genes in PD pathogenesis suggesting it as a new potential therapeutic target. Resveratrol, a natural polyphenolic compound proved to improve mitochondrial activity through the activation of several metabolic sensors resulting in PGC-1α activation. Here we have tested in vitro the effect of resveratrol treatment on primary fibroblast cultures from two patients with early-onset PD linked to different Park2 mutations. We show that resveratrol regulates energy homeostasis through activation of AMP-activated protein kinase (AMPK) and sirtuin 1 (SIRT1) and raise of mRNA expression of a number of PGC-1α's target genes resulting in enhanced mitochondrial oxidative function, likely related to a decrease of oxidative stress and to an increase of mitochondrial biogenesis. The functional impact of resveratrol treatment encompassed an increase of complex I and citrate synthase activities, basal oxygen consumption, and mitochondrial ATP production and a decrease in lactate content, thus supporting a switch from glycolytic to oxidative metabolism. Moreover, resveratrol treatment caused an enhanced macro-autophagic flux through activation of an LC3-independent pathway. Our results, obtained in early-onset PD fibroblasts, suggest that resveratrol may have potential clinical application in selected cases of PD-affected patients

Single-cell profiling of myasthenia gravis identifies a pathogenic T cell signature.

Myasthenia gravis (MG) is an autoimmune disease characterized by impaired neuromuscular signaling due to autoantibodies targeting the acetylcholine receptor. Although its auto-antigens and effector mechanisms are well defined, the cellular and molecular drivers underpinning MG remain elusive. Here, we employed high-dimensional single-cell mass and spectral cytometry of blood and thymus samples from MG patients in combination with supervised and unsupervised machine-learning tools to gain insight into the immune dysregulation underlying MG. By creating a comprehensive immune map, we identified two dysregulated subsets of inflammatory circulating memory T helper (Th) cells. These signature ThCD103 and ThGM cells populated the diseased thymus, were reduced in the blood of MG patients, and were inversely correlated with disease severity. Both signature Th subsets rebounded in the blood of MG patients after surgical thymus removal, indicative of their role as cellular markers of disease activity. Together, this in-depth analysis of the immune landscape of MG provides valuable insight into disease pathogenesis, suggests novel biomarkers and identifies new potential therapeutic targets for treatment