44 research outputs found
Conditions determining PCR success in otter landscape genetics
No full textNon-invasive sampling for genetic analysis using DNA from feces is a useful technique to monitor animal populations. But despite being a very promising tool, it still has some limitations and can give some problems such as low success rates of genotyped samples, contamination concerns and high microsatellite genotyping error rates. To optimize success, it is important to follow specific steps, including a stringent sample selection in the field, proper storage conditions, and choice of the optimal DNA extraction method for the study species. Furthermore, the PCR success rate could potentially be influenced by factors prior to collection, such as temperature and the diet of the species, as well as by sample storage time. Here, we investigated the effect of temperature at time of collection, average maximum temperature one week prior to collection and storage time on the amplification success rate of microsatellite DNA extracted from XX fecal samples of the Eurasian otter (Lutra lutra) in the Netherlands. To do this, we considered two cases: firstly, we performed a logistic regression to assess whether temperature at time of collection and/or storage time could be used to predict the genotyping success rate for one microsatellite locus, that is used as a primary selection tool for sample quality. Secondly, we considered PCR success rate as the number of successfully scored loci out of the total 13 loci used in the landscape genetic study of Dutch otters and assessed the effect of temperature at time of collection, average maximum temperatures one week prior to collection and storage time. Results showed that when considering one locus only, the amplification success rate was not influenced by temperature. However, when considering all the loci temperature at time of collection and temperature one week prior positively influenced the PCR success rate, meaning that as the temperature raised, the success rate increased. This result was contrasting most of the literature, indeed, as it would be expected, high temperatures should cause a more rapid degradation of DNA caused by higher activity of hydrolytic enzymes in feces. Probably there were outside factors (e.g. the otters’ diet) that we did not control and that were confounded with temperature requiring the need for further investigation. Storage time had no effect on DNA amplification success rate in any of our tests. Our results suggest that DNA amplification success rate is influenced by many factors. Before starting a non-invasive study, it is fundamental to take into account several factors and to study to what extent each factor influences the amplification success in order to maximize the DNA amplification success rate and to reduce genotyping error rates
Monitoreo genético no invasivo de una población de nutrias silvestres en los Países Bajos.
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Evaluation of the effectiveness of neuromuscular electrical stimulation after total knee arthroplasty: a meta-analysis
No full textObjective: The aim of the study was to evaluate the efficacy of the use of the neuromuscular electrical stimulation after total knee arthroplasty. Design: The study used a systematic review of randomized controlled trials (MEDLINE, PubMed, Cochrane Library, and PEDro) using Patient Population or Problem, Intervention, Comparison, Outcomes, Setting approach to formulate the research question, controlled terms, and Boolean operators. Inclusion and exclusion criteria were defined in advance. "Neuromuscular electrical stimulation" and "total knee arthroplasty" were used as keywords. The overall risk of bias was determined according to the following: random sequence generation, concealment, blinding mass of participants and staff, commissioning blind assessment results, incomplete data, and loans received. Results: Of the 36 identified studies, six were included in the review (496 participants). In these studies, one group of patients followed a rehabilitation protocol (control group) and the other followed a rehabilitation program plus a session of neuromuscular electrical stimulation (neuromuscular electrical stimulation group). Patients of neuromuscular electrical stimulation groups got the best scores (timed up and go test, stair climbing test, and walk test). Neuromuscular electrical stimulation benefits were strong in the first postoperative weeks/months and gradually diminished. Conclusions: Neuromuscular electrical stimulation allows a slightly better functional recovery after total knee arthroplasty, especially in the first period, with more evident benefits in patients with a severe lack of muscular activation. Nevertheless, there is no difference at medium-long term
Immunotherapy and the Combination with Targeted Therapies for Advanced Hepatocellular Carcinoma
No full textOne of the most important abilities of a tumor is to establish a state of immunosuppression inside the tumor microenvironment. This is made possible through numerous mechanisms of tumor immune escape that have been identified in experimental studies during the last decades. In addition, the hepatic microenvironment is commonly oriented towards a state of immune tolerance because the liver receives blood from the hepatic arteries and portal veins containing a variety of endogenous antigens. Therefore, the hepatic microenvironment establishes an autoimmune tolerance, preventing an autoimmune reaction in the liver. On this basis, hepatic tumor cells may escape the immune system, avoiding being recognized and destroyed by immune cells. Moreover, since the etiology of Hepatocellular Carcinoma (HCC) is often related to cirrhosis, and hepatitis B or C, this tumor develops in the context of chronic inflammation. Thus, the HCC microenvironment is characterized by important immune cell infiltration. Given these data and the poor prognosis of advanced HCC, different immunotherapeutic strategies have been developed and evaluated for these patients. In this review, we describe all the clinical applications of immunotherapy for advanced HCC, from the drugs that have already been approved to the ongoing clinical trials
