The Efficacy of Selected Sodium Hypochlorite Heating Methods for Increasing and Maintaining Its Intracanal Temperature—An Ex Vivo Study

Background: Enhancement of the temperature of sodium hypochlorite (NaOCl) solution would increase its cleaning potential and decontamination of the root canal system. Therefore, the aim of the present in vitro investigation was to compare the efficacy of different methods of NaOCl heating by evaluating the temperature profiles developed at different levels of the root canal system. Methods: Five thermocouples were applied at different levels of the root canal system of extracted human premolars. NaOCl solution was heated according to two methods: extraoral heating (50 °C, 60 °C, and 70 °C) using a magnetic hotplate heater and intracanal heating by F-06, XF-30/04, and ML-12 pluggers at 100 °C, 150 °C, and 180 °C. Results: The extraoral heating method was ineffective to produce a significant temperature increase at the root apex. Comparable results were obtained using the intracanal heating method through the ML-12 plugger that showed slightly better results only when set at 180 °C. On the other hand, negligible differences were observed in terms of temperature maintenance at several levels of the root between the F-06 and XF-30/04 pluggers, even though the time intervals were higher in case of XF-30/04. Conclusions: The intracanal heating method provided a better temperature persistence in the middle third of the root canal system. Conversely, extraoral heating was ineffective to produce a significant temperature increase at the apex of the root. Comparable results were obtained even using the ML-12 plugger

Differential inhibition of the TGF-b signaling pathway in HCC cells using the small molecule inhibitor LY2157299 and the D10 monoclonal antibody against TGF-β receptor Type II

We investigated blocking the TGF-b signaling pathway in HCC using two small molecule inhibitors (LY2157299, LY2109761) and a neutralizing humanized antibody (D10) against TGF-bRII. LY2157299 and LY2109761 inhibited HCC cell migration on Laminin-5, Fibronectin, Vitronectin, Fibrinogen and Collagen-I and de novo phosphorylation of pSMAD2. LY2157299 inhibited HCC migration and cell growth independently of the expression levels of TGF-bRII. In contrast to LY2157299, D10 showed a reduction in pSMAD2 only after a short exposure. This study supports the use of LY2157299 in clinical trials, and presents new insights into TGF-b receptor cycling in cancer cells

2C-Cas9: a versatile tool for clonal analysis of gene function

CRISPR/Cas9-mediated targeted mutagenesis allows efficient generation of loss-of-function alleles in zebrafish. To date this technology has been primarily used to generate genetic knockout animals. Nevertheless, the study of the function of certain loci might require tight spatiotemporal control of gene inactivation. Here, we show that tissue-specific gene disruption can be achieved by driving Cas9 expression with the Gal4/UAS system. Furthermore, by combining the Gal4/UAS and Cre/loxP systems, we establish a versatile tool to genetically label mutant cell clones, enabling their phenotypic analysis. Our technique has the potential to be applied to diverse model organisms, enabling tissue-specific loss-of-function and phenotypic characterization of live and fixed tissues

Platelet-rich plasma in arthroscopic repairs of complete tears of the rotator cuff

OBJETIVO: Avaliar os resultados funcionais e o índice de rerrotura do reparo do manguito rotador por via artroscópica associado ao uso do PRP. MÉTODOS: Série de casos prospectiva, avaliando os resultados do reparo artroscópico do manguito rotador em fileira simples associada ao uso do PRP. Foram incluídas apenas roturas isoladas do supraespinal, com retração inferior a 3cm. O PRP utilizado foi obtido pelo método de aférese, e aplicado em sua forma ativada, com a adição de trombina autóloga, na consistência líquida. A avaliação pós-operatória foi realizada de maneira padronizada, aos 12 meses de seguimento. Foram utilizadas as escalas de Constant-Murley, UCLA e EVA, além da análise da incidência de rerroturas através da ressonância magnética. RESULTADOS: Foram avaliados 14 pacientes (14 ombros). A escala de Constant-Murley evoluiu em média de 45,64 ± 12,29 no pré-operatório para 80,78 ± 13,22 no pós-operatório (p < 0,001). A escala de UCLA sofreu um incremento de 13,78 ± 5,66 para 31, 43 ± 3,9 (p < 0,001). A dor dos pacientes apresentou uma melhora significativa de acordo com a EVA (p = 0,0013), decrescendo de uma mediana de 7,5 (p25% = 6, p75% = 8) para 0,5 (p25% = 0, p75% = 3). Nenhum dos pacientes apresentou rerrotura completa. Em três pacientes (21,4%) foi observada uma rerrotura parcial, sem transfixação. Apenas um paciente evoluiu com complicação (capsulite adesiva). CONCLUSÃO: Os pacientes submetidos ao reparo do manguito rotador por via artroscópica associado ao uso do PRP apresentaram uma melhora funcional significativa e nenhuma rerrotura completa

Neutralizing antibodies to Omicron after the fourth SARS-CoV-2 mRNA vaccine dose in immunocompromised patients highlight the need of additional boosters

IntroductionImmunocompromised patients have been shown to have an impaired immune response to COVID-19 vaccines.MethodsHere we compared the B-cell, T-cell and neutralizing antibody response to WT and Omicron BA.2 SARS-CoV-2 virus after the fourth dose of mRNA COVID-19 vaccines in patients with hematological malignancies (HM, n=71), solid tumors (ST, n=39) and immune-rheumatological (IR, n=25) diseases. The humoral and T-cell responses to SARS-CoV-2 vaccination were analyzed by quantifying the anti-RBD antibodies, their neutralization activity and the IFN-γ released after spike specific stimulation.ResultsWe show that the T-cell response is similarly boosted by the fourth dose across the different subgroups, while the antibody response is improved only in patients not receiving B-cell targeted therapies, independent on the pathology. However, 9% of patients with anti-RBD antibodies did not have neutralizing antibodies to either virus variants, while an additional 5.7% did not have neutralizing antibodies to Omicron BA.2, making these patients particularly vulnerable to SARS-CoV-2 infection. The increment of neutralizing antibodies was very similar towards Omicron BA.2 and WT virus after the third or fourth dose of vaccine, suggesting that there is no preferential skewing towards either virus variant with the booster dose. The only limited step is the amount of antibodies that are elicited after vaccination, thus increasing the probability of developing neutralizing antibodies to both variants of virus.DiscussionThese data support the recommendation of additional booster doses in frail patients to enhance the development of a B-cell response directed against Omicron and/or to enhance the T-cell response in patients treated with anti-CD20

Mortality from gastrointestinal congenital anomalies at 264 hospitals in 74 low-income, middle-income, and high-income countries: a multicentre, international, prospective cohort study

Summary Background Congenital anomalies are the fifth leading cause of mortality in children younger than 5 years globally. Many gastrointestinal congenital anomalies are fatal without timely access to neonatal surgical care, but few studies have been done on these conditions in low-income and middle-income countries (LMICs). We compared outcomes of the seven most common gastrointestinal congenital anomalies in low-income, middle-income, and high-income countries globally, and identified factors associated with mortality. Methods We did a multicentre, international prospective cohort study of patients younger than 16 years, presenting to hospital for the first time with oesophageal atresia, congenital diaphragmatic hernia, intestinal atresia, gastroschisis, exomphalos, anorectal malformation, and Hirschsprung’s disease. Recruitment was of consecutive patients for a minimum of 1 month between October, 2018, and April, 2019. We collected data on patient demographics, clinical status, interventions, and outcomes using the REDCap platform. Patients were followed up for 30 days after primary intervention, or 30 days after admission if they did not receive an intervention. The primary outcome was all-cause, in-hospital mortality for all conditions combined and each condition individually, stratified by country income status. We did a complete case analysis. Findings We included 3849 patients with 3975 study conditions (560 with oesophageal atresia, 448 with congenital diaphragmatic hernia, 681 with intestinal atresia, 453 with gastroschisis, 325 with exomphalos, 991 with anorectal malformation, and 517 with Hirschsprung’s disease) from 264 hospitals (89 in high-income countries, 166 in middleincome countries, and nine in low-income countries) in 74 countries. Of the 3849 patients, 2231 (58·0%) were male. Median gestational age at birth was 38 weeks (IQR 36–39) and median bodyweight at presentation was 2·8 kg (2·3–3·3). Mortality among all patients was 37 (39·8%) of 93 in low-income countries, 583 (20·4%) of 2860 in middle-income countries, and 50 (5·6%) of 896 in high-income countries (p<0·0001 between all country income groups). Gastroschisis had the greatest difference in mortality between country income strata (nine [90·0%] of ten in lowincome countries, 97 [31·9%] of 304 in middle-income countries, and two [1·4%] of 139 in high-income countries; p≤0·0001 between all country income groups). Factors significantly associated with higher mortality for all patients combined included country income status (low-income vs high-income countries, risk ratio 2·78 [95% CI 1·88–4·11], p<0·0001; middle-income vs high-income countries, 2·11 [1·59–2·79], p<0·0001), sepsis at presentation (1·20 [1·04–1·40], p=0·016), higher American Society of Anesthesiologists (ASA) score at primary intervention (ASA 4–5 vs ASA 1–2, 1·82 [1·40–2·35], p<0·0001; ASA 3 vs ASA 1–2, 1·58, [1·30–1·92], p<0·0001]), surgical safety checklist not used (1·39 [1·02–1·90], p=0·035), and ventilation or parenteral nutrition unavailable when needed (ventilation 1·96, [1·41–2·71], p=0·0001; parenteral nutrition 1·35, [1·05–1·74], p=0·018). Administration of parenteral nutrition (0·61, [0·47–0·79], p=0·0002) and use of a peripherally inserted central catheter (0·65 [0·50–0·86], p=0·0024) or percutaneous central line (0·69 [0·48–1·00], p=0·049) were associated with lower mortality. Interpretation Unacceptable differences in mortality exist for gastrointestinal congenital anomalies between lowincome, middle-income, and high-income countries. Improving access to quality neonatal surgical care in LMICs will be vital to achieve Sustainable Development Goal 3.2 of ending preventable deaths in neonates and children younger than 5 years by 2030

Edition du génome pour comprendre la formation des circuits neuronaux : une nouvelle stratégie CRSPR/Cas9 pour la mutagenèse conditionnelle et étude fonctionnelle du rôle de la famille des gènes des météorines dans le développement neurologique du poisson zébra

In recent years, the zebrafish (Danio rerio) has emerged as a powerful model organism to study neuronal circuit development and function. To date, different genome editing technologies allow the generation of constitutive mutant alleles, permitting the study of gene loss-of-function in this vertebrate model. Nevertheless, to assess the role of certain loci it might be required a precise spatiotemporal control of gene inactivation. The rst part of my thesis describes a novel strategy for tissue-specific gene disruption based on the CRISPR/Cas9 and the Gal4/UAS systems. The described technique allows the induction of somatic mutations in genetically labeled tissues, cell clones or single cells, making it possible to follow the effect of gene disruption in vivo via reporter gene expression. The second part of the thesis focuses on the functional analysis of the role of the meteorin gene family during neuronal development and axonal targeting in zebra sh. Meteorin family is conserved among vertebrates and its members have been shown to be involved in neuronal progenitor proliferation and differentiation and axonal elongation, in vitro. We used the zebrafish nervous system as a model to dissect the role of Meteorins during embryonic development, focusing on their potential role as novel guidance molecules. Interestingly, we found that genes belonging to the meteorin family are expressed along the midline of the larval central nervous system and at the floor plate in the hindbrain and spinal cord. We generated CRISPR/Cas9 mutant lines carrying out-of-frame deletions in the coding sequence of each member of the zebrafish meteorin family and we performed a comprehensive analysis of the establishment of axonal projections in the mutants. Our data pointed out that metrns loss-of-function affects the earliest process of axonal development, demonstrating a crucial role in the process of axonal outgrowth for this new family of evolutionary conserved guidance molecules.Depuis quelques années, le poisson zèbre (Danio rerio) est devenu un modèle de choix pour l'étude du système nerveux et de ses fonctions. Récemment, des technologies nouvelles d'édition du génome permettent la génération d'allèles mutés de manière constitutionnelle et l'étude fonctionnelle de gènes chez ce modèle vertébré. Néanmoins, certains loci nécessite une inactivation spatiotemporelle précise et contrôlée. La première partie de ma thèse décrit la mise au point d'une nouvelle stratégie de disruption génétique de manière tissu-spécifique, basée sur la technologie du CRISPR/Cas9 et du système UAS/Gal4. Cette technique permet l'introduction de mutations somatiques dans des tissus, des clones ou des cellules individuelles préalablement génétiquement marqués, rendant ainsi possible le suivi in vivo de l'effet de la mutation générée grâce au gène rapporteur. La seconde partie de ma thèse se centre sur l'étude fonctionnelle d'une famille des gènes, les meteorines, durant le développement du système nerveux et lors du ciblage axonale chez le poisson zèbre. Les Meteorines sont des protéines conservées chez les vertébrés qui ont été impliquées dans la prolifération, la différentiation des progéniteurs de neurones et notamment dans l'élongation axonale in vitro. Nous avons pu mettre en évidence que les meteorines sont exprimées le long de la ligne médiane du système nerveux chez les larves et au niveau du plancher de la partie postérieure du cerveau et de la moelle épinière. Par l'utilisation du CRISPR/Cas9, nous avons généré des lignées mutantes pour chaque gène meteorine et avons ainsi procédé à l'analyse de l'établissement des projections axonales dans ces lignées mutantes

Icoxilòpoli 2. Iconografia delle xilografie del Polifilo

Abstract en (english translation by Flavia De Nicola revised by Giulia Martina Weston) In Venice, in 1499, Aldo Manuzio the Elder gave the most important incunabulum of the Italian Renaissance to the press: the Hypnerotomachia Poliphili, accompanied by 171 woodcuts. Over the years, many studies have followed one another to look into the identity of the anonymous author, but only a few of them were dedicated to the 171 images illustrating this printed book, and nowadays the name of the draftsman and that of the engraver remain uncertain. In 2017, the research project Icoxilòpoli 1 - Iconographies of the Polifilo’s woodcuts, led by Stefano Colonna in the context of the Museology and Art Criticism and Restoration teaching at the University of Rome La Sapienza, was founded on the ascertainment that specific symbolic themes of the incunabulum are described in the text but not through the images, and vice versa, hence the need to identify the keywords and the symbolic objects both in the text and images, in order to create a two-way path "word-image-word". Icoxilòpoli 2, which takes up Icoxilòpoli 1 subject matters, presents 19 insets relating to Polifilo’s woodcuts, ordered by number of appearance in the editio princeps of the Hypnerotomachia dating 1499 and equipped with further analytical essays. The authors well understood the need to continue to operate on an expressive double-track, which could encourage an interpretation of the insets in the key of a “metafictional” story while preserving the academic status of the scientific approach, and this shows how rich Italian academia is in human and scientific potentialities so that, if enhanced, it produces academic results since the first years of research. Abstract it Nel 1499, a Venezia, Aldo Manuzio il Vecchio dava alle presse il più importante incunabolo del rinascimento italiano: l’Hypnerotomachia Poliphili, corredato di ben 171 xilografie. Negli anni si sono susseguiti molti studi in merito a chi ne fosse l’anonimo autore, ma solo pochi sono stati dedicati alle 171 immagini che illustrano questo libro stampato, e ancora oggi restano incerti il nome del disegnatore e quello dell’incisore. Nel 2017 nasce il Progetto di ricerca Icoxilòpoli 1 - Iconografie delle xilografie del Polifilo, diretto da Stefano Colonna nel suo Insegnamento di Museologia e Critica Artistica e del restauro della Sapienza, fondato sulla constatazione che determinati argomenti simbolici dell’incunabolo sono descritti nel testo ma non nelle immagini, e viceversa, e quindi sulla necessità di individuare, sia nel testo che nelle immagini, le “parole-chiave” e gli oggetti simbolici, per creare un percorso biunivoco “parola-immagine-parola”. Icoxilòpoli 2, che riprende le tematiche di Icoxilòpoli 1, presenta 19 schede relative alle xilografie del Polifilo, ordinate secondo il numero di apparizione nell’editio princeps dell’Hypnerotomachia del 1499 con il corredo di ulteriori saggi analitici. Gli autori hanno ben compreso la necessità di rimanere su un doppio binario espressivo, che favorisse una lettura delle schede in chiave di “meta-romanzo” pur conservando la qualifica accademica dell’approccio scientifico, e ciò dimostra quanto l’università italiana sia ricca di potenzialità umane e scientifiche che, se valorizzate, producono risultati accademici fin dai primi anni di studio