Does the anaesthetic influence behavioural transmission of the monogenean Gyrodactylus gasterostei Glaser, 1974 off the host?

The aim of this study was to investigate the use of the anaesthetic 2-phenoxyethanol on the transmission factors of gyrodactylid and to ascertain how this may affect in the colonisation of new hosts using the Gyrodactylus gasterostei Glaser, 1974 - Gasterosteus aculeatus L. model which is a simple and successful system to examine aspects of transmission of parasites from live and dead fish. Laboratory experiments include determining the maturity (presence of male copulate organ) and reproductive (presence of daughter) status of transmitting worms, in order to consider the factors that influence parasite option to migrate to a new individual of the same host species. This study demonstrates that parasites with a Male Copulate Organ (MCO) present are more likely to abandon the host and attempt a host transfer. The use of the anaesthetic 2-phenoxyethanol does not affect transmission of gyrodactylids which leave the host to colonise a new host. Finally, the use of other anaesthetic although its relative importance with respect to transmission remains uncertain.Department of the Environment, Food and Rural Affairs (Defra); Overseas Research Students Awards Scheme (ORSAS) UK; Consejo Nacional Ciencia y Tecnologia, Mexico CONACyT [171032]info:eu-repo/semantics/publishedVersio

A large-scale empirical exploration on refactoring activities in open source software projects

Refactoring is a well-established practice that aims at improving the internal structure of a software system without changing its external behavior. Existing literature provides evidence of how and why developers perform refactoring in practice. In this paper, we continue on this line of research by performing a large-scale empirical analysis of refactoring practices in 200 open source systems. Specifically, we analyze the change history of these systems at commit level to investigate: (i) whether developers perform refactoring operations and, if so, which are more diffused and (ii) when refactoring operations are applied, and (iii) which are the main developer-oriented factors leading to refactoring. Based on our results, future research can focus on enabling automatic support for less frequent refactorings and on recommending refactorings based on the developer's workload, project's maturity and developer's commitment to the project

Predicting intention and maintenance of breastfeeding up to 2-years after birth in primiparous and multiparous women

Objectives: Breastfeeding up to 2-years has been associated with short and long-term health benefits for both newborns and mothers. However, few women breastfeed up to 2-years after birth. This study extends previous research on the theory of planned behaviour (TPB) examining the predictors of intention and maintenance of breastfeeding up to 2-years in both primiparous and multiparous women. Design: 155 pregnant women participated in this longitudinal study. Methods: Expectant mothers completed a questionnaire and then 2-years after the child’s birth were asked to report breastfeeding behaviour. Interactions among parity and TPB constructs were examined. Results: Attitudes, descriptive and injunctive norms, and perceived behavioural control (PBC) explained 58% of the variance in mothers’ intention to breastfeed. Attitudes were the strongest predictor, followed by PBC, descriptive norms and parity. A significant interaction was found between parity and PBC, showing that PBC was only a significant predictor of intention to breastfeed at 2-years in multiparous women. Intentions predicted breastfeeding behaviour at 2-years. Conclusion: Promoting intentions may be a useful way to increase breastfeeding duration to 2-years and targeting attitudes and norms may be one way to increase intentions. Further, targeting PBC may also be useful to increase intentions, but only in multiparous women

UN ENFOQUE EXPERIMENTAL SOBRE EL COMPORTAMIENTO EN LA TRANSMISIÓN DE GYRODACTYLUS (MONOGENEA) FUERA DEL HOSPEDERO 'EL PEZ ESPINOSO' (GASTEROSTEUS ACULEATUS L.)

El objetivo de este estudio fue investigar algunos factores en el comportamiento durante la transmisión del monogeneo Gyrodactylus sp. y determinar si la madurez y estado reproductivo del parásito pueden ser empleados en la colonización de nuevos hospederos. El modelo de transmisión empleado fue Gyrodactylus gasterostei Glaser, 1974 y G. arcuatus Bychowsky 1933 - Gasterosteus aculeatus L. que migraron de peces sacrificados. Los experimentos en el laboratorio incluyen la determinación de la madurez (presencia del órgano copulador masculino) y reproductiva (presencia de la hija en útero), y determinar los factores de comportamiento que influyen en la elección de los parásitos para migrar a un nuevo hospedero. Este estudio es el primer enfoque experimental en el comportamiento con relación a diferentes etapas de desarrollo en dos especies de Gyrodactylus utilizando el mismo modelo de hospedero. El presente trabajo examinó los aspectos iniciales del comportamiento de las estrategias de comportamiento en gyrodactilidos aislados de forma individual. Los resultados muestran que la comparación de las frecuencias de comportamiento entre los individuos que sobrevivieron al registro de observaciones y el análisis de los datos no mostró diferencias de comportamiento entre estas especies de monogeneos

Osteoblasts Display Different Responsiveness to TRAIL-Induced Apoptosis During Their Differentiation Process

Apoptosis can occur throughout the life span of osteoblasts (OBs), beginning from the early stages of differentiation and continuing throughout all stages of their working life. Here, we investigated the effects of tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) on normal human OBs showing for the first time that the expression of TRAIL receptors is modulated during OB differentiation. In particular, the TRAIL receptor ratio was in favor of the deaths because of the low expression of DcR2 in undifferentiated OBs, differently it was shifted toward the decoys in differentiated ones. Undifferentiated OBs treated with TRAIL showed reduced cell viability, whereas differentiated OBs displayed TRAIL resistance. The OB sensitiveness to TRAIL was due to the up-regulation of DR5 and the down-regulation of DcR2. The main death receptor involved in TRAIL-reduced OB viability was DR5 as demonstrated by the rescue of cell viability observed in the presence of anti-DR5 neutralizing antibody. Besides the ratio of TRAIL receptors, the sensitivity of undifferentiated OBs to TRAIL-cytotoxic effect was also associated with low mRNA levels of intracellular anti-apoptotic proteins, such as cFLIP, the activation of caspase-8 and -3, as well as the DNA fragmentation. This study suggests that apoptotic effect exerted by TRAIL/TRAIL-receptor system on normal human OB is strictly dependent upon cell differentiation status

Branch coverage prediction in automated testing

This is the peer reviewed version which has been published in final form at [DOI]. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions.Software testing is crucial in continuous integration (CI). Ideally, at every commit, all the test cases should be executed, and moreover, new test cases should be generated for the new source code. This is especially true in a Continuous Test Generation (CTG) environment, where the automatic generation of test cases is integrated into the continuous integration pipeline. In this context, developers want to achieve a certain minimum level of coverage for every software build. However, executing all the test cases and, moreover, generating new ones for all the classes at every commit is not feasible. As a consequence, developers have to select which subset of classes has to be tested and/or targeted by test‐case generation. We argue that knowing a priori the branch coverage that can be achieved with test‐data generation tools can help developers into taking informed decision about those issues. In this paper, we investigate the possibility to use source‐code metrics to predict the coverage achieved by test‐data generation tools. We use four different categories of source‐code features and assess the prediction on a large data set involving more than 3'000 Java classes. We compare different machine learning algorithms and conduct a fine‐grained feature analysis aimed at investigating the factors that most impact the prediction accuracy. Moreover, we extend our investigation to four different search budgets. Our evaluation shows that the best model achieves an average 0.15 and 0.21 MAE on nested cross‐validation over the different budgets, respectively, on EVOSUITE and RANDOOP. Finally, the discussion of the results demonstrate the relevance of coupling‐related features for the prediction accuracy

LIGHT as regulator of bone homeostasis during osteolytic bone metastasis formation in non-small cell lung cancer patients

Tumor necrosis factor superfamily member 14 (TNFSF14), LIGHT is one of the cytokines produced by tumor and immune cells, which promotes homeostasis of lymphoid organs, liver and bone. Nonsmall cell lung cancer (NSCLC) commonly metastasizes bone, altering bone homeostasis and causing osteolysis. Here we investigated the role of LIGHT in NSCLC-induced osteolytic bone disease. The LIGHT expression in monocytes was higher in patients with metastatic bone lesions than in non-bone metastatic ones (66.5 ± 24.5 vs 43.3 ± 25.2 mean ± SD, p = 0.001), in healthy donors (66.5 ± 24.5 vs 8.5 ± 4.6 p = 0.0002), and in non-bone metastatic patients than in healthy donors (43.3 ± 25.2 vs 8.5 ± 4.6, p = 0.0001). Serum LIGHT levels were also significantly higher in bone metastatic patients than in non-bone metastatic ones (186.8 ± 191.2 pg/ml vs 115.8 ± 73 pg/ml, p = 0.04) and in healthy donors (186.8 ± 191.2 pg/ml vs 85.7 ± 38.4 pg/ml, p = 0.04). A neutralizing mAb anti-LIGHT added to osteoclast (OC) cultures of both bone and non-bone metastases inhibited osteoclastogenesis, but the decrease was statistically significant only for bone metastatic patients (272 ± 98 vs 132 ± 74, p = 0.01). To investigate the role of LIGHT in NSCLC- induced bone lesion in vivo, we performed an intratibial injection of a mouse lung cancer cell line LLC-1, in wild-type (WT) and LIGHT KO mice. The WT-injected mice displayed a significant reduction of about 20% for BV/TV, Tb.N, Tb.Th, and Tb.Sp compared to the WT-vehicle mice (pb 0.01). These parameters did not show significant variation for KO-injected mice vs vehicle or for WT-injected mice vs KO-injected mice. These data indicate LIGHT as a regulator of bone homeostasis during NSCLC metastatic invasion, thus it may be a novel therapeutic target in osteolytic bone metastases

Irisin, the novel myokine responsible for benefits of physical exercise on bone

It has been recently reported that, after physical exercise activity, the skeletal muscle releases Irisin, the newly identified myokine able of driving transition of white adipocytes into brown, following a phenomenon known as the browning response. This result suggested that skeletal muscle is crucial in the regulation of energy homeostasis, supporting its role as endocrine organ that targets adipose tissue by promoting energy expenditure. In accordance with this new finding, we demonstrated that conditioned media (CM) collected from primary myoblasts of exercised mice were able to induce osteoblast differentiation in a greater extent than those of mice housed in resting conditions and this effect is Irisin-mediated. In view of further proving the involvement of Irisin in bone metabolism, we validate its direct effect on osteoblasts by using r-Irisin. Here we show that phosphorylation of MAP kinase ERK and expression of Atf 4 (p<0,001), the key trascription factor of osteoblast differentiation, were significantly increased after Irisin treatment. Furthermore, ALP and pro-Collagen I mRNA resulted up regulated (p<0,001), as we already demonstrated by treating osteoblasts with conditioned medium from primary myoblasts of exercised mice. To recapitulate in vivo the effect of physical exercise, we injected mice with r-Irisin. Our results show that BV/TV of Irisin-treated mice was higher than vehicle-injected mice. In elderly, the severe decline of skeletal muscle function, known as Sarcopenia, is associated with impaired function of bone (Osteopenia) and these two simultaneous losses of function lead to increased risk of bone fractures. In order to reveal new strategies for treatment of sarcopenia and osteopenia, we also analyzed the effect of physical activity in old mice. Our findings demonstrate that mRNA levels of the most relevant bone proteins resulted up regulated in ex-vivo osteoblast obtained from exercised old mice compared with mice kept in resting conditions. Our data highlight a novel link in muscle-fat-bone axis demonstrating that Irisin targets bone tissue directly. Future perspectives, based on these studies, could satisfy the ongoing research of exercise-mimetic therapies with anabolic action on the skeleton

Palmitate-induced lipotoxicity alters acetylation of multiple proteins in clonal β cells and human pancreatic islets

Type 2 diabetes is characterized by progressive β cell dysfunction, with lipotoxicity playing a possible pathogenetic role. Palmitate is often used to examine the direct effects of lipotoxicity and it may cause mitochondrial alterations by activating protein acetylation. However, it is unknown whether palmitate influences protein acetylation in β cells. We investigated lysine acetylation in mitochondrial proteins from INS-1E β cells (INS-1E) and in proteins from human pancreatic islets (HPI) after 24 h palmitate exposure. First, we confirmed that palmitate damages β cells and demonstrated that chemical inhibition of deacetylation also impairs INS-1E function and survival. Then, by 2-D gel electrophoresis, Western Blot and Liquid Chromatography-Mass Spectrometry we evaluated the effects of palmitate on protein acetylation. In mitochondrial preparations from palmitate-treated INS-1E, 32 acetylated spots were detected, with 13 proteins resulting over-acetylated. In HPI, 136 acetylated proteins were found, of which 11 were over-acetylated upon culture with palmitate. Interestingly, three proteins, glutamate dehydrogenase, mitochondrial superoxide dismutase, and SREBP-1, were over-acetylated in both INS-1E and HPI. Therefore, prolonged exposure to palmitate induces changes in β cell protein lysine acetylation and this modification could play a role in causing β cell damage. Dysregulated acetylation may be a target to counteract palmitate-induced β cell lipotoxicity

Osteogenic differentiation of dental follicle stem cells.

Background: Stem cells are defined as clonogenic cells capable of self-renewal and multi-lineage differentiation. A population of these cells has been identified in human Dental Follicle (DF). Dental Follicle Stem Cells (DFSCs) were found in pediatric unerupted wisdom teeth and have been shown to differentiate, under particular conditions, into various cell types of the mesenchymal tissues. Aim: The aim of this study was to investigate if cells isolated from DF show stem features, differentiate toward osteoblastic phenotype and express osteoblastic markers. Methods: We studied the immunophenotype of DFSCs by flow cytometric analysis, the osteoblastic markers of differentiated DFSCs were assayed by histochemical methods and real-time PCR. Results: We demonstrated that DFSCs expressed a heterogeneous assortment of makers associated with stemness. Moreover DFSCs differentiated into osteoblast-like cells, producing mineralized matrix nodules and expressed the typical osteoblastic markers, Alkaline Phosphatase (ALP) and Collagen I (Coll I). Conclusion: This study suggests that DFSCs may provide a cell source for tissue engineering of bone