Gene targeting in adult rhesus macaque fibroblasts

<p>Abstract</p> <p>Background</p> <p>Gene targeting in nonhuman primates has the potential to produce critical animal models for translational studies related to human diseases. Successful gene targeting in fibroblasts followed by somatic cell nuclear transfer (SCNT) has been achieved in several species of large mammals but not yet in primates. Our goal was to establish the protocols necessary to achieve gene targeting in primary culture of adult rhesus macaque fibroblasts as a first step in creating nonhuman primate models of genetic disease using nuclear transfer technology.</p> <p>Results</p> <p>A primary culture of adult male fibroblasts was transfected with hTERT to overcome senescence and allow long term <it>in vitro </it>manipulations. Successful gene targeting of the HPRT locus in rhesus macaques was achieved by electroporating S-phase synchronized cells with a construct containing a SV40 enhancer.</p> <p>Conclusion</p> <p>The cell lines reported here could be used for the production of null mutant rhesus macaque models of human genetic disease using SCNT technology. In addition, given the close evolutionary relationship and biological similarity between rhesus macaques and humans, the protocols described here may prove useful in the genetic engineering of human somatic cells.</p

Gene-enhanced tissue engineering for dental hard tissue regeneration: (1) overview and practical considerations

Gene-based therapies for tissue regeneration involve delivering a specific gene to a target tissue with the goal of changing the phenotype or protein expression profile of the recipient cell; the ultimate goal being to form specific tissues required for regeneration. One of the principal advantages of this approach is that it provides for a sustained delivery of physiologic levels of the growth factor of interest. This manuscript will review the principals of gene-enhanced tissue engineering and the techniques of introducing DNA into cells. Part 2 will review recent advances in gene-based therapies for dental hard tissue regeneration, specifically as it pertains to dentin regeneration/pulp capping and periodontal regeneration

P1 receptors and cytokine secretion

Evidence has accumulated in the last three decades to suggest tissue protection and regeneration by adenosine in multiple different cell types. Adenosine produced in hypoxic or inflamed environments reduces tissue injury and promotes repair by receptor-mediated mechanisms. Among other actions, regulation of cytokine production and secretion by immune cells, astrocytes and microglia (the brain immunocytes) has emerged as a main mechanism at the basis of adenosine effects in diseases characterized by a marked inflammatory component. Many recent studies have highlighted that signalling through A1 and A2A adenosine receptors can powerfully prevent the release of pro-inflammatory cytokines, thus inhibiting inflammation and reperfusion injury. However, the activation of adenosine receptors is not invariably protective of tissues, as signalling through the A2B adenosine receptor has been linked to pro-inflammatory actions which are, at least in part, mediated by increased release of pro-inflammatory cytokines from epithelial cells, astrocytes and fibroblasts. Here, we discuss the multiple actions of P1 receptors on cytokine secretion, by analyzing, in particular, the role of the various adenosine receptor subtypes, the complex reciprocal interplay between the adenosine and the cytokine systems, their pathophysiological significance and the potential of adenosine receptor ligands as new anti-inflammatory agents

Sex-Differences in the Pattern of Comorbidities, Functional Independence, and Mortality in Elderly Inpatients: Evidence from the RePoSI Register

Background: The RePoSi study has provided data on comorbidities, polypharmacy, and sex dimorphism in hospitalised elderly patients. Methods: We retrospectively analysed data collected from the 2010, 2012, 2014, and 2016 data sets of the RePoSi register. The aim of this study was to explore the sex-differences and to validate the multivariate model in the entire dataset with an expanded follow-up at 1 year. Results: Among 4714 patients, 51% were women and 49% were men. The disease distribution showed that diabetes, coronary artery disease, chronic obstructive pulmonary disease, chronic kidney disease, and malignancy were more frequent in men but that hypertension, anaemia, osteoarthritis, depression, and diverticulitis disease were more common in women. Severity and comorbidity indexes according to the Cumulative Illness Rating Scale (CIRS-s and CIRS-c) were higher in men, while cognitive impairment, mood disorders, and disability in daily life measured by the Barthel Index (BI) were worse in women. In the multivariate analysis, BI, CIRS, and malignancy significantly increased the risk of death in men at the 1-year follow-up, while age was independently associated with mortality in women. Conclusions: Our study highlighted the relevance and the validity of our previous predictive model in the identification of sex dimorphism in hospitalised elderly patients underscoring the need of sex-personalised health-care

Statins as a new therapeutic perspective in myocarditis and postmyocarditis dilated cardiomyopathy

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Disruption of the CNTF gene results in motor neuron degeneration

CNTF is a cytosolic molecule expressed postnatally in myelinating Schwann cells and in a subpopulation of astrocytes. Although CNTF administration prevents lesion-mediated and genetically determined motor neuron degeneration, its physiological function remained elusive. Here it is reported that abolition of CNTF gene expression by homologous recombination results in a progressive atrophy and loss of motor neurons in adult mice, which is functionally reflected by a small but significant reduction in muscle strength

Surveillance for severe acute respiratory infections in the 2019/2020 season in Tuscany, Italy

Influenza is a major public health burden. In Italy there were 7.6 million symptomatic case of influenza in the 2019/2020 influenza season. In Italy, the influenza season lasts from October to April of the following year. We analysed influenza A and B viruses from hospitalized patients with Severe Acute Respiratory Infections (SARI) to carry out epidemiological and virological surveillance

Surveillance for severe acute respiratory infections in the 2019/2020 season in Tuscany, Italy

Abstract Background Influenza is a major public health burden. In Italy there were 7.6 million symptomatic case of influenza in the 2019/2020 influenza season. In Italy, the influenza season lasts from October to April of the following year. We analysed influenza A and B viruses from hospitalized patients with Severe Acute Respiratory Infections (SARI) to carry out epidemiological and virological surveillance. Methods 68 oropharyngeal swabs were taken from the patients with SARI at Le Scotte University Hospital in Siena and they were given a questionnaire to record their underlying disease and vaccination status. Total RNA was extracted from the swabs by means of the QIAamp Viral RNA Mini kit and RT-PCR was carried out. After, biodirectional DNA sequencing reactions were performed. All statistical analyses were performed by means of GraphPad Prism 6 software. Results The median age of subject was 82 years and 52.9% were female. The subjects showed fever (89.7%), fatigue (77%), headache (47%), cough (75%), sore throat (70.5%), breathlessness (63.2%).We found that 20.5% of 68 subject were positive (13% for A H3N2 and 7% for A H1N1). Out of the 68 subject, 25% received seasonal influenza vaccination (91.6% trivalent vaccine and 8.4% quadrivalent vaccine). The most common underlying disease found in the positive subjects were cardiovascular diseases (50%), renal diseases (50%), lung diseases (42.8%), diabetes (35.7%), cancer (35%). Conclusions Epidemiological and virological surveillance remains an essential tool for monitoring virus circulation and possible mismatches with seasonal vaccine strains, and providing information that can be used to improve the composition of influenza vaccines. Our data support the importance of seasonal vaccination in subjects with chronic diseases to reduce hospitalisation and mortality. Key messages Virological surveillance is important for determining the spread of influenza viruses. Seasonal vaccination is considered as the most effective way to prevent influenza and its complications. </jats:sec