Quantitative Polymerase Chain Reaction (qPCR) is currently the most sensitive technique used for absolute and relative quantification of a target gene transcript, requiring the use of appropriated ...reference genes for data normalization. To accurately estimate the relative expression of target tomato (Solanum lycopersicum L.) genes responsive to several virus species in reverse transcription qPCR analysis, the identification of reliable reference genes is mandatory. In the present study, ten reference genes were analyzed across a set of eight samples: two tomato contrasting genotypes ('Santa Clara', susceptible, and its near-isogenic line 'LAM 157', resistant); subjected to two treatments (inoculation with Tomato chlorotic mottle virus (ToCMoV) and its mock-inoculated control) and in two distinct times after inoculation (early and late). Reference genes stability was estimated by three statistical programs (geNorm, NormFinder and BestKeeper). To validate the results over broader experimental conditions, a set of ten samples, corresponding to additional three tomato-virus pathosystems that included tospovirus, crinivirus and tymovirus + tobamovirus, was analyzed together with the tomato-ToCMoV pathosystem dataset, using the same algorithms. Taking into account the combined analyses of the ranking order outputs from the three algorithms, TIP41 and EF1 were identified as the most stable genes for tomato-ToCMoV pathosystem, and TIP41 and EXP for the four pathosystems together, and selected to be used as reference in the forthcoming expression qPCR analysis of target genes in experimental conditions involving the aforementioned tomato-virus pathosystems.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Throughout recent years, ungulates have experienced significant increases in numbers and geographic range sizes in Europe, becoming locally overabundant populations in some regions. Changes in ...legislation regarding poaching, abandonment of land and re‐naturalisation of habitats, and decreasing numbers of hunters, among other things, have led to alarming scenarios in wild ungulate biology. Although ungulates bring some financial benefits for ecosystems and society through tourism and hunting, the problems associated with populations that are no longer controlled can outweigh the advantages. Damage to forestry and agriculture, ungulate‐vehicle collisions, and diseases are among the most concerning problems related to ungulate overabundance.
To deal with these problems and to decide on the best management strategy to apply, it is essential to have tools available to monitor populations with an integrative approach based on ecological change indicators, and to assess population and ecosystem status. Furthermore, in a globalised world, people’s opinions matter, and sociological studies regarding human perception of wild mammals must take place in order to allow proper management, including consideration of people’s expectations as well as animal and ecosystem needs. Successful and unsuccessful management strategies have already been attempted, and the knowledge of consequences over time enables an adaptive approach.
Management of ungulate populations is a complex subject, and each case should be studied, analysing the cost_performance balance of measures to be taken, and ensuring ongoing financial means to carry out and continue with successful ecosystem management strategies. Multidisciplinary teams should be built, including biologists, veterinarians, stakeholders, sociologists, and others, to deal with the management of European wild ungulate populations.
Ungulates such as red deer and wild boar have been increasing in the past decades in Europe, and so have their consequences. Damage to agriculture and forestry, emerging infectious diseases, and increasing ungulate‐vehicle collisions are only some of the consequences of ungulate populations that are no longer controlled. Ungulate expansion also increases conflicts among humans, and the study of these interactions should be integrated in an holistic perspective of ecosystems. This review gives researchers and the general public a brief summary of the situation of ungulates in Europe, suggests a novel socio‐ecological approach, and provides recommendations for stakeholders on how to deal with this situation, which is likely to intensify in the future.
Pediatric acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL) exhibit favorable survival rates. However, for AML and ALL patients carrying KMT2A gene translocations clinical outcome ...remains unsatisfactory. Key players in KMT2A-fusion-driven leukemogenesis include menin and DOT1L. Recently, menin inhibitors like revumenib have garnered attention for their potential therapeutic efficacy in treating KMT2A-rearranged acute leukemias. However, resistance to menin inhibition poses challenges, and identifying which patients would benefit from revumenib treatment is crucial. Here, we investigated the in vitro response to revumenib in KMT2A-rearranged ALL and AML. While ALL samples show rapid, dose-dependent induction of leukemic cell death, AML responses are much slower and promote myeloid differentiation. Furthermore, we reveal that acquired resistance to revumenib in KMT2A-rearranged ALL cells can occur either through the acquisition of MEN1 mutations or independently of mutations in MEN1. Finally, we demonstrate significant synergy between revumenib and the DOT1L inhibitor pinometostat in KMT2A-rearranged ALL, suggesting that such drug combinations represent a potent therapeutic strategy for these patients. Collectively, our findings underscore the complexity of resistance mechanisms and advocate for precise patient stratification to optimize the use of menin inhibitors in KMT2A-rearranged acute leukemia.
The Iberian wolf (Canis lupus signatus) is a top predator that inhabits the Iberian Peninsula. In Portugal, its numbers and distribution declined throughout the 20th century, due to human ...persecution, habitat degradation and prey decline, which have led to higher predation rates of livestock in the remaining packs. In Montesinho Natural Park (northeast Portugal), wild ungulate populations have been increasing in the last years, which may have led wolf to predate upon them. In order to assess Iberian wolf diet in this area, 85 wolf scats were collected from transects distributed throughout the study area in two periods between November 2017 and August 2019. Scat analysis indicated a high predation on wild ungulates, where the frequency of occurrence showed that roe deer was the most consumed prey (44%), followed by red deer (26%) and wild boar (24%). Domestic/wild cat (6%), domestic goat and stone marten (5%) were consumed in lower quantities. It was found a higher selection towards roe deer (D = 0.71) and this was the only prey item which was significantly dependent of the season of the year (χ2 = 16.95, df = 3, p < 0.001). This is the first study in Portugal where was recorded that wolves feed mainly on wild ungulates. We conclude that lower livestock predation may be correlated with higher wild ungulates densities in our study area, as well as suitable husbandry practices, leading to a shift on Iberian wolf diet from mainly livestock on previous studies to wild ungulates.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
The eastern Brazilian Amazon contains many isolated ferruginous savanna ecosystem patches (locally known as 'canga vegetation') located on ironstone rocky outcrops on the top of plateaus and ridges, ...surrounded by tropical rainforests. In the Carajás Mineral Province (CMP), these outcrops contain large iron ore reserves that have been exploited by opencast mining since the 1980s. The canga vegetation is particularly impacted by mining, since the iron ores that occur are associated with this type of vegetation and currently, little is known regarding the extent of canga vegetation patches before mining activities began. This information is important for quantifying the impact of mining, in addition to helping plan conservation programmes. Here, land cover changes of the Canga area in the CMP are evaluated by estimating the pre-mining area of canga patches and comparing it to the actual extent of canga patches. We mapped canga vegetation using geographic object-based image analysis (GEOBIA) from 1973 Landsat-1 MSS, 1984 and 2001 Landsat-5 TM, and 2016 Landsat-8 OLI images, and found that canga vegetation originally occupied an area of 144.2 km2 before mining exploitation. By 2016, 19.6% of the canga area was lost in the CMP due to conversion to other land-use types (mining areas, pasturelands). In the Carajás National Forest (CNF), located within the CMP, the original canga vegetation covered 105.2 km2 (2.55% of the CNF total area), and in 2016, canga vegetation occupied an area of 77.2 km2 (1.87%). Therefore, after more than three decades of mineral exploitation, less than 20% of the total canga area was lost. Currently, 21% of the canga area in the CMP is protected by the Campos Ferruginosos National Park. By documenting the initial extent of canga vegetation in the eastern Amazon and the extent to which it has been lost due to mining operations, the results of this work are the first step towards conserving this ecosystem.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Summary Background Incorrect use of inhaler devices remains an obstacle for respiratory diseases management. We aimed to evaluate the frequency of inhaler technique errors; to determine the devices ...perceived as the easiest and favourite to use; to study the association of device type, demographics and patient preferences with inhaler technique (IT). Methods Cross-sectional assessment of 301 adults, with asthma (194) or chronic pulmonary obstructive disease, undergoing treatment with Aerolizer® , Autohaler® , Breezehaler® , Diskus® , Handihaler® , MDI without spacer, Miat-haler® , Novolizer® , Respimat® and/or Turbohaler® . Patients completed self-assessment questionnaires and face-to-face interview, with demonstration of inhaler technique. The rate of wrong steps (number of wrong steps ÷ number of total steps; RWS) was the primary outcome. Adjusted odds ratio (aOR) (95% confidence intervals CI) for presenting ≥1 IT errors were computed. Results From the 464 inhaler technique performances, the median RWS was 18%. Turbohaler® (21%) and Diskus® (19%) were chosen as easiest and Novolizer® (18%), Diskus® (18%), Turbohaler® (17%) as favourite for daily use. Females (aOR 2.68 95% CI 1.55–4.65; vs. males, patients with >64 yr (aOR 2.73 95% CI 1.15–6.48; vs <45 yr) and patients using Aerolizer® or Handihaler® (aOR 3.24 95% CI 1.13–9.32 and aOR 3.71 95% CI 1.38–10.2, respectively) were more likely to perform IT errors; otherwise, no association was found, including with using the favourite device (aOR 1.43 95% CI 0.84–2.42). Conclusion The frequency of inhaler technique errors was high and no device was clearly preferred over the others. Using the preferred inhaler device was not associated with less errors.
The human dimension of wildlife is a subject of increasing interest, especially considering the potential impact of people's perceptions on decision-making concerning wildlife management and thus on ...species' distribution and abundance. This is particularly important for species that inhabit human-dominated landscapes, where conflicts are likely to arise. These conflicts typically emerge between different human collectives when their interests collide and thus an assessment of their perceptions is valuable. Throughout Europe, ungulates have expanded, and Portugal is no exception. The expansion of red deer (Cervus elaphus) comes with benefits (e.g. hunting opportunities), but also with costs (e.g. vehicle collision or damage to crops), that can shape people's perceptions of these populations. To assess perceptions of red deer populations in continental Portugal, we developed a questionnaire survey with three interest groups: general public, farmers and hunters (total n = 1532). Our results show that perceptions about red deer were generally positive with a high acknowledgement of deer benefits, which we link to a broad level of sympathy towards this species. In addition, farmers showed a higher concern with crop damage caused by deer than non-farmers. Nonetheless, a general lack of knowledge regarding wildlife was present, particularly among younger people, who are potentially more apart and disconnected from nature. Now is the time to implement preventive and mitigation measures – e.g. fencing, reduction of population density – which can be a small contribution to address this problem. The focus should be put on people's acceptance of the required strategies to manage deer populations (which can also include unpopular practices, such as hunting). For that, an evaluation of people's attitudes towards deer populations and awareness of associated problems is essential. People's opinions, as well as the success of monitoring and management strategies, should be evaluated through multi-disciplinary teams, that include natural and social scientists, to ensure their success.
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•General perception about red deer was better than expected.•Farmers presented the highest damage perception.•Hunter had high interest in wildlife but acknowledge deer damages.•Strategies to ensure acceptance of generally unpopular management measures are needed.•Multidisciplinary teams should be built to solve wildlife management topics.
Background
Refined phenotyping of allergic diseases may unravel novel phenotypes. Conjunctivitis as an independent disorder has never been approached.
Aim
To identify distinct classes of allergic ...respiratory diseases using latent class analysis (LCA) and distinguish each class using classification and regression tree (CART) analysis.
Methods
Seven hundred and twenty‐eight adults from the Portuguese general population study ICAR had a structured medical interview combined with blood collection, skin prick tests, spirometry with bronchodilation, and exhaled nitric oxide. LCA was applied to 19 variables. The CART algorithm selected the most likely variables distinguishing LCA‐classes.
Results
A six‐class model was obtained. Class 1 (25%): nonallergic participants without bronchial or ocular symptoms. Classes 2 (22%) and 3 (11%): nasal and ocular (low levels) symptoms without nasal impairment, monosensitized (Class 2) or polysensitized (Class 3). Class 4 (13%): polysensitized participants with high levels of nasal and ocular symptoms, and nasal impairment. Classes 5 (16%) and 6 (14%): high level of nasal, bronchial and ocular symptoms with nasal impairment (non‐allergic or polysensitized, respectively). Participants in classes 5 and 6 had more bronchial exacerbations and unscheduled medical visits (P < 0.001). Ocular symptoms were significantly higher in classes with nasal impairment, compared to those without impairment (P < 0.001) or no nasal symptom (P < 0.001). CART highlighted ocular symptoms as the most relevant variable in distinguishing LCA‐classes.
Conclusion
Novel severe phenotypes of participants with co‐occurrence of ocular, nasal and bronchial symptoms, and exacerbation‐prone were identified. The tree algorithm showed the importance of the ocular symptoms in the expression of allergic diseases phenotypes.
Novel phenotypes of allergic respiratory diseases were identified by latent class analysis, in adults from a general population sample. For the first time, ocular symptoms were used independently and ranked by classification and regression tree analysis as the most relevant variable to separate those phenotypes.
Nanoparticles (NPs), as drug delivery systems, appear to be a promising tool for prolonged therapeutic strategies as they allow a controlled drug release over time. However, most of the studies found ...in the literature simply contemplate the use of a single or low number of dosages with low NPs concentrations. In the context of chronic diseases, like Alzheimer's disease, cancer or human immunodeficiency virus (HIV), where the therapeutic scheme is also chronic, studies with numerous repeated dosages are often neglected.
We screened different NPs, polymeric and lipid-based, in a repeated-dose toxicity study, to evaluate the safety and tissue distribution of promising nanocarriers to be used in the treatment of long-lasting diseases.
After administrating 24 high concentrated doses of the selected NPs intraperitoneally (i.p.) (3 times a week for 2 months), animals have presented NPs accumulation in different tissues. However, neither toxicity, bodyweight changes nor clinical signs of disease were observed.
This work demonstrates no general adverse effects upon the studied NPs repeated-dose exposure, indicating the most promising NPs to be used in the different therapeutic circumstances, which may be useful in chronic diseases treatment.
Previous research in rodents suggests that the long‐term neurobehavioral disturbances induced by chronic ethanol (EtOH) exposure could be due to endocannabinoid system (ECS) alterations. Moreover, ...ECS failure has been proposed to mediate the cognitive impairment and β‐amyloid production in Alzheimer disease (AD). Thus, in the present study, we evaluated the effects of adolescent EtOH binge drinking on the cognitive disturbances, hippocampal β‐amyloid levels, and in the ECS expression on a transgenic mouse model (APP/PSEN, AZ) of AD. We exposed AZ and wild‐type mice to a binge‐drinking treatment during adolescence. At 6 and 12 months of age, we evaluated hippocampal‐dependent learning and memory: β‐amyloid concentrations and RNA and protein levels of cannabinoid type‐2 receptors (CB2), diacylglycerol lipase‐α (DAGLα), and monoacylglycerol lipase (MAGL) in the hippocampus. The results showed that binge‐EtOH treatment worsens cognitive function and increases β‐amyloid levels in AZ. At 6 months, EtOH heightens CB2 (RNA and protein) and DAGLα (RNA) expression in wild type but not in AZ. On the contrary, EtOH enhances MAGL RNA expression only in AZ. At 12 months, AZ displays increased levels of CB2 (RNA and protein) and DAGLα (protein) compared with control. Similar to what happens at 6 months, EtOH induces an increase in CB2 gene expression in wild type but not in AZ; however, it augments CB2 and DAGLα protein levels in both genotypes. Therefore, we propose that adolescent binge drinking accelerates cognitive deficits associated with aging and AD. It also accelerates hippocampal β‐amyloid accumulation in AZ and affects differently the ECS response in wild type and AZ.
In the present research we demonstrate that exposure to ethanol binge‐drinking during all the adolescent period accelerates cognitive deterioration, enhances hippocampal β‐amyloid production and differently alters the endocannabinoid system (ECS) signaling in a rodent model of Alzheimer's disease (AD) compared to wild‐type mice. We propose that these ECS alterations could be a factor that contributes to the aggravation of AD neuropathology in our AZ mice exposed to EtOH.
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