A secondary RET mutation in the activation loop conferring resistance to vandetanib

Resistance to vandetanib, a type I RET kinase inhibitor, developed in a patient with metastatic lung adenocarcinoma harboring a CCDC6-RET fusion that initially exhibited a response to treatment. The resistant tumor acquired a secondary mutation resulting in a serine-to-phenylalanine substitution at codon 904 in the activation loop of the RET kinase domain. The S904F mutation confers resistance to vandetanib by increasing the ATP affinity and autophosphorylation activity of RET kinase. A reduced interaction with the drug is also observed in vitro for the S904F mutant by thermal shift assay. A crystal structure of the S904F mutant reveals a small hydrophobic core around F904 likely to enhance basal kinase activity by stabilizing an active conformer. Our findings indicate that missense mutations in the activation loop of the kinase domain are able to increase kinase activity and confer drug resistance through allosteric effects

Examining a staging model for anorexia nervosa: empirical exploration of a four stage model of severity.

Background: An illness staging model for anorexia nervosa (AN) has received increasing attention, but assessing the merits of this concept is dependent on empirically examining a model in clinical samples. Building on preliminary findings regarding the reliability and validity of the Clinician Administered Staging Instrument for Anorexia Nervosa (CASIAN), the current study explores operationalising CASIAN severity scores into stages and assesses their relationship with other clinical features. Method: In women with DSM-IV-R AN and sub-threshold AN (all met AN criteria using DSM 5), receiver operating curve (ROC) analysis (n = 67) assessed the relationship between the sensitivity and specificity of each stage of the CASIAN. Thereafter chi-square and post-hoc adjusted residual analysis provided a preliminary assessment of the validity of the stages comparing the relationship between stage and treatment intensity and AN sub-types, and explored movement between stages after six months (Time 3) in a larger cohort (n = 171). Results: The CASIAN significantly distinguished between milder stages of illness (Stage 1 and 2) versus more severe stages of illness (Stages 3 and 4), and approached statistical significance in distinguishing each of the four stages from one other. CASIAN Stages were significantly associated with treatment modality and primary diagnosis, and CASIAN Stage at Time 1 was significantly associated with Stage at 6 month follow-up. Conclusions: Provisional support is provided for a staging model in AN. Larger studies with longer follow-up of cases are now needed to replicate and extend these findings and evaluate the overall utility of staging as well as optimal staging models

Multiple endocrine neoplasia type 2 syndromes (MEN 2): results from the ItaMEN network analysis on the prevalence of different genotypes and phenotypes.

OBJECTIVE: Multiple endocrine neoplasia type 2 (MEN 2) is a genetic disease characterized by medullary thyroid carcinoma (MTC) associated (MEN 2A and 2B) or not familial MTC (FMTC) with other endocrine neoplasia due to germline RET gene mutations. The prevalence of these rare genetic diseases and their corresponding RET mutations are unknown due to the small size of the study population. METHODS: We collected data on germline RET mutations of 250 families with hereditary MTC followed in 20 different Italian centres. RESULTS AND CONCLUSIONS: The most frequent RET amino acid substitution was Val804Met (19.6%) followed by Cys634Arg (13.6%). A total of 40 different germline RET mutations were present. Six families (2.4%) were negative for germline RET mutations. The comparison of the prevalence of RET germline mutations in the present study with those published by other European studies showed a higher prevalence of Val804Met and Ser891Ala mutations and a lower prevalence of Leu790Phe and Tyr791Phe (P<0.0001). A statistically significant higher prevalence of mutations affecting non-cysteine codons was also found (P<0.0001). Furthermore, the phenotype data collection showed an unexpected higher prevalence of FMTC (57.6%) with respect to other MEN 2 syndromes (34% MEN 2A and 6.8% of MEN 2B). In conclusion, we observed a statistically significant different pattern of RET mutations in Italian MEN 2 families with respect to other European studies and a higher prevalence of FMTC phenotype. The different ethnic origins of the patients and the particular attention given to analysing apparently sporadic MTC for RET germline mutations may explain these findings

Essential oils against Varroa destructor: a soft way to fight the parasitic mite of Apis mellifera

Essential oils (EOs) extracted from the aromatic plants Artemisia annua, Artemisia verlotiorum, Cinnamomum verum, and Citrus reticulata were investigated as repellents against the honey bees parasitic mite Varroa destructor. In laboratory tests, all EOs except C. reticulata exerted significant repellent activity against the mite after 24 h exposure. C. verumwas the most effective EO against V. destructor (median effective concentration EC50 =1.30 μL L−1), and the least toxic against honey bees (EC50 = 13.29 μL L−1). Because of its high selectivity ratio (A. mellifera LD50/V. destructor EC50 = 10.22), C. verum EO was then tested to control varroosis in colonies in field trials. The results of open field tests confirmed the efficacy observed in the laboratory. After one week of treatment, colonies treated with the EO showed a significant reduction of V. destructor infestation (about 65% at the dose of 25.0 μL L−1) and no negative effects on A. mellifera. Overall, our experiment indicated that C. verum EO could be used to effectively control varroosis in the hive with no side effects on the bee colonies

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Multi-biomarker approach and IBR index to evaluate the effects of different contaminants on the ecotoxicological status of Apis mellifera

The honeybee, Apis mellifera L. (Hymenoptera: Apidae), a keystone pollinator of wild plant species and agricultural crops, is disappearing globally due to parasites and diseases, habitat loss, genetic constraints, beekeeper management issues and to the widespread use of pesticides. Besides insecticides, widely studied in this species, honeybees are also exposed to herbicides and fungicides and heavy metals whose lethal and sublethal effects need to be investigated. In this context, our study aimed to evaluate the effects of fungicides and of heavy metals on honeybees and to develop and apply a multi-biomarker approach that include an Integrated Biological Index (IBRv2) to assess the toxicological status of this species. Biomarkers of neurotoxicity (AChE and CaE), metabolic alteration (ALP, and GST) and immune system (LYS, granulocytes) were measured, following honeybees’ exposure to cadmium or to a crop fungicide, using the genotoxic compound EMS as positive control. A biomarker of genotoxicity (NA assay) was developed and applied for the first time in honeybees. At the doses tested, all the contaminants showed sublethal toxicity to the bees, highlighting in particular genotoxic effects. The data collected were analyzed by an IBRv2 index, which integrated the seven biomarkers used in this study. IBRv2 index increased with increasing cadmium or fungicide concentrations. The IBRv2 represents a simple tool for a general description of honeybees ecotoxicological health status. Results highlight the need for more in-depth investigations on the effects of fungicides on non-target organisms, such as honeybees, using sensitive methods for the determination of sublethal effects. This study contributes to the development of a multi-biomarker approach to be used for a more accurate ecotoxicological environmental monitoring of these animals

A multibiomarker approach to evaluate the impact of anthropogenic contaminants on the ecotoxicological status of honey bees, Apis mellifera

A rapid decline of Apis mellifera, a keystone pollinator of wild plant species and agricultural crops, was recorded worldwide in recent years. The massive use of insecticides and fungicides in agriculture associated with pollution generated by other human activities and presence of parasites can cause toxicological effects in bees including a decrease of the immune defences, leading to the collapse of the colonies. Effective assessment of the ecotoxicological impacts of anthropogenic contaminants requires an approach that combines different biomarkers that enable a more precise diagnosis of exposure to environmental stressors through a combination of different biological responses. The aim of this study was to develop and apply a set of biomarkers to study the ecotoxicological status of honey bees. In the first phase, we investigated in the laboratory the effects of EMS, cadmium and a commercial fungicide (azoxistrobin 18.2% and ciproconazole 7.3%) in adult honey bees, evaluating eventual variation in glutathione S-transferase (GST), carboxylesterase (CaE), acetylcholinesterase (AChE) activities, alkaline phosphatase (ALP), lysozyme, erythrocyte nuclear abnormalities (ENA) assay and differential haemocytes count (DHC). Genotoxic effects, as well as alteration of the immune system, were found in bees treated with EMS, cadmium or the fungicide. Cadmium and the fungicide also inhibited AChE and CaE activities, GST was induced by all the compounds investigated. In the second phase, adult honey bees were collected from apiaries located in four environments characterized by different chemical input: a wooded environment (low input), an urban site, an orchard and a cultivated countryside site. Honey bees from the urban site were also collected and analyzed before and after treatment for parasites. ENA assay showed that bees taken from the countryside and the orchard had a greater number of abnormalities compared to the forest, confirming the presence of genotoxic substances in agricultural environments compared to control environments. GST activity was induced in bees from the urban environment, AChE was inhibited in the countryside compared to the forest, suggesting the presence of substances with neurotoxic effect in this environment. ALP activity was induced in all sites in comparison to wooded one. The bees collected after the parasites treatment showed an increase for GST activity as well as AChE inhibition

Cigarette smoking, nicotine dependence and anxiety disorders : a systematic review of population-based, epidemiological studies

Background Multiple studies have demonstrated that rates of smoking and nicotine dependence are increased in individuals with anxiety disorders. However, significant variability exists in the epidemiological literature exploring this relationship, including study design (cross-sectional versus prospective), the population assessed (random sample versus clinical population) and diagnostic instrument utilized.Methods We undertook a systematic review of population-based observational studies that utilized recognized structured clinical diagnostic criteria (Diagnostic and Statistical Manual of Mental Disorders (DSM) or International Classification of Diseases (ICD)) for anxiety disorder diagnosis to investigate the relationship between cigarette smoking, nicotine dependence and anxiety disorders.Results In total, 47 studies met the predefined inclusion criteria, with 12 studies providing prospective information and 5 studies providing quasiprospective information. The available evidence suggests that some baseline anxiety disorders are a risk factor for initiation of smoking and nicotine dependence, although the evidence is heterogeneous and many studies did not control for the effect of comorbid substance use disorders. The identified evidence however appeared to more consistently support cigarette smoking and nicotine dependence as being a risk factor for development of some anxiety disorders (for example, panic disorder, generalized anxiety disorder), although these findings were not replicated in all studies. A number of inconsistencies in the literature were identified.Conclusions Although many studies have demonstrated increased rates of smoking and nicotine dependence in individuals with anxiety disorders, there is a limited and heterogeneous literature that has prospectively examined this relationship in population studies using validated diagnostic criteria. The most consistent evidence supports smoking and nicotine dependence as increasing the risk of panic disorder and generalized anxiety disorder. The literature assessing anxiety disorders increasing smoking and nicotine dependence is inconsistent. Potential issues with the current literature are discussed and directions for future research are suggested

Essential oils as post-harvest crop protectants against the fruit fly drosophila suzukii: Bioactivity and organoleptic profile

The essential oils extracted from mandarin (Citrus reticulata Blanco) fruits, and from tea tree (Maleleuca alternifolia (Maiden and Betche) Cheel) leaves have been chemically analyzed and tested for their bioactivity against D. suzukii. Besides, to estimate consumers’ acceptability of the essential oil (EO) treatments, we evaluated their impact on the organoleptic characteristics of the EO-treated fruits. The main chemical constituents of the two EOs were 1,8-cineole and 4-terpineol for M. alternifolia (22.4% and 17.6% of the total components, respectively), and limonene (83.6% of the total components) for C. reticulata. The behavioral tests indicate that the two EOs are able to deter D. suzukii oviposition and that D. suzukii shows positive chemotaxis to low concentrations of the EOs and negative chemotaxis when the EO concentration increases. While no negative effects on the organoleptic profiles were detected for fruits treated with C. reticulata EO, the olfactory profile of fruits treated with M. alternifolia EO was so negative that they were defined as “not suitable for consumption” by panellists. Overall, our findings indicate that the use of EOs for the post-harvest protection of small fruits is feasible, provided that the essential oils are selected not only for their bioactivity against the insect pest but also for their affinity with the consumers’ sensorial system

Widefield fluorescence imaging as an auxiliary tool to select the biopsy site for actinic cheilitis diagnosis

Actinic cheilitis (AC) is considered a potentially malignant disorder that mainly affects the lower lip, and it is caused by prolonged sun exposure. Clinical diagnosis relies on visual inspection by a trained clinician, when suspected of dysplasia changes, a biopsy is required. The heteregenous characteristics of the AC, makes the choice of the biopsy site a difficult task. Fluorescence detection has been presented as a useful tool to to detect biochemical and morphological tissue features related to cancer diagnosis, but still its effectiveness to discriminate premalignant lesion is not completely defined. In this clinical study, 57 AC patients were investigated using widefield fluorescence imaging (WFI) to evaluate the efficacy of this technique as an auxiliary tool to biopsy site location. A handheld fluorescence system based on 400-450 nm LED illumination Distinct trained clinicians evaluate the patient either with the conventional examination or the WFI, and were blinded to the other evaluation. A biopsy site was chosen based on the clinical examination, and another site was chosen using the fluorescence visualization. A total of 114 punch biopsies were performed, and 93% of the tissue samples presented epithelial dysplasia. The majority of the sites that presented moderate or severe dysplasia were sites chosen by WFI, showing its efficiency to improve the diagnosis of AC.CNPq (475322/2011-8)CEPID-CePOF/FAPESP (98/14270-8