Simultaneous brain, brainstem and spinal cord pharmacological-fMRI reveals involvement of an endogenous opioid network in attentional analgesia

Pain perception is decreased by shifting attentional focus away from a threatening event. This attentional analgesia engages parallel descending control pathways from anterior cingulate (ACC) to locus coeruleus, and ACC to periaqueductal grey (PAG) - rostral ventromedial medulla (RVM), indicating possible roles for noradrenergic or opioidergic neuromodulators. To determine which pathway modulates nociceptive activity in humans we used simultaneous whole brain-spinal cord pharmacological-fMRI (N=39) across three sessions. Noxious thermal forearm stimulation generated somatotopic-activation of dorsal horn (DH) whose activity correlated with pain report and mirrored attentional pain modulation. Activity in an adjacent cluster reported the interaction between task and noxious stimulus. Effective connectivity analysis revealed that ACC interacts with PAG and RVM to modulate spinal cord activity. Blocking endogenous opioids with Naltrexone impairs attentional analgesia and disrupts RVM-spinal and ACC-PAG connectivity. Noradrenergic augmentation with Reboxetine did not alter attentional analgesia. Cognitive pain modulation involves opioidergic ACC-PAG-RVM descending control which suppresses spinal nociceptive activity

Structural and non-coding variants increase the diagnostic yield of clinical whole genome sequencing for rare diseases

BACKGROUND: Whole genome sequencing is increasingly being used for the diagnosis of patients with rare diseases. However, the diagnostic yields of many studies, particularly those conducted in a healthcare setting, are often disappointingly low, at 25-30%. This is in part because although entire genomes are sequenced, analysis is often confined to in silico gene panels or coding regions of the genome.METHODS: We undertook WGS on a cohort of 122 unrelated rare disease patients and their relatives (300 genomes) who had been pre-screened by gene panels or arrays. Patients were recruited from a broad spectrum of clinical specialties. We applied a bioinformatics pipeline that would allow comprehensive analysis of all variant types. We combined established bioinformatics tools for phenotypic and genomic analysis with our novel algorithms (SVRare, ALTSPLICE and GREEN-DB) to detect and annotate structural, splice site and non-coding variants.RESULTS: Our diagnostic yield was 43/122 cases (35%), although 47/122 cases (39%) were considered solved when considering novel candidate genes with supporting functional data into account. Structural, splice site and deep intronic variants contributed to 20/47 (43%) of our solved cases. Five genes that are novel, or were novel at the time of discovery, were identified, whilst a further three genes are putative novel disease genes with evidence of causality. We identified variants of uncertain significance in a further fourteen candidate genes. The phenotypic spectrum associated with RMND1 was expanded to include polymicrogyria. Two patients with secondary findings in FBN1 and KCNQ1 were confirmed to have previously unidentified Marfan and long QT syndromes, respectively, and were referred for further clinical interventions. Clinical diagnoses were changed in six patients and treatment adjustments made for eight individuals, which for five patients was considered life-saving.CONCLUSIONS: Genome sequencing is increasingly being considered as a first-line genetic test in routine clinical settings and can make a substantial contribution to rapidly identifying a causal aetiology for many patients, shortening their diagnostic odyssey. We have demonstrated that structural, splice site and intronic variants make a significant contribution to diagnostic yield and that comprehensive analysis of the entire genome is essential to maximise the value of clinical genome sequencing.</p

Phylogenetic ctDNA analysis depicts early-stage lung cancer evolution.

The early detection of relapse following primary surgery for non-small-cell lung cancer and the characterization of emerging subclones, which seed metastatic sites, might offer new therapeutic approaches for limiting tumour recurrence. The ability to track the evolutionary dynamics of early-stage lung cancer non-invasively in circulating tumour DNA (ctDNA) has not yet been demonstrated. Here we use a tumour-specific phylogenetic approach to profile the ctDNA of the first 100 TRACERx (Tracking Non-Small-Cell Lung Cancer Evolution Through Therapy (Rx)) study participants, including one patient who was also recruited to the PEACE (Posthumous Evaluation of Advanced Cancer Environment) post-mortem study. We identify independent predictors of ctDNA release and analyse the tumour-volume detection limit. Through blinded profiling of postoperative plasma, we observe evidence of adjuvant chemotherapy resistance and identify patients who are very likely to experience recurrence of their lung cancer. Finally, we show that phylogenetic ctDNA profiling tracks the subclonal nature of lung cancer relapse and metastasis, providing a new approach for ctDNA-driven therapeutic studies

Discutindo a educação ambiental no cotidiano escolar: desenvolvimento de projetos na escola formação inicial e continuada de professores

A presente pesquisa buscou discutir como a Educação Ambiental (EA) vem sendo trabalhada, no Ensino Fundamental e como os docentes desta escola compreendem e vem inserindo a EA no cotidiano escolar., em uma escola estadual do município de Tangará da Serra/MT, Brasil. Para tanto, realizou-se entrevistas com os professores que fazem parte de um projeto interdisciplinar de EA na escola pesquisada. Verificou-se que o projeto da escola não vem conseguindo alcançar os objetivos propostos por: desconhecimento do mesmo, pelos professores; formação deficiente dos professores, não entendimento da EA como processo de ensino-aprendizagem, falta de recursos didáticos, planejamento inadequado das atividades. A partir dessa constatação, procurou-se debater a impossibilidade de tratar do tema fora do trabalho interdisciplinar, bem como, e principalmente, a importância de um estudo mais aprofundado de EA, vinculando teoria e prática, tanto na formação docente, como em projetos escolares, a fim de fugir do tradicional vínculo “EA e ecologia, lixo e horta”.Facultad de Humanidades y Ciencias de la Educació

COAST (Cisplatin Ototoxicity Attenuated by Aspirin Trial): A phase II double blind, randomised controlled trial to establish if aspirin reduces cisplatin induced hearing loss

BackgroundCisplatin has the highest ototoxic potential of platinum containing drugs leaving 50% of patients with a permanent and irreversible hearing-loss, and associated reduction in quality of life. There are no preventative treatment strategies to minimise this common side-effect. Both cisplatin and gentamicin are thought to damage hearing through a common mechanism involving reactive oxygen species in the inner ear. Aspirin has been shown to minimise gentamicin induced ototoxicity. We tested the hypothesis that aspirin could reduce ototoxicity from cisplatin based chemotherapy.Method94 patients were recruited into a phase II, double blind, placebo controlled, 2-arm trial, and randomised 1:1 from multiple tumour-sites (head and neck, thoracic, bladder, germ cell) in the U.K. Patients underwent pure tone audiometry (PTA) before, and following their final cisplatin dose at 7 and 90 days. Patients received aspirin 975mg tid and omeprazole 20 mg od, or placebos; from the day before, to 2 days after their cisplatin dose, for each cisplatin cycle. The primary endpoint was total hearing difference, comparing pre- and post-cisplatin PTA at 6 and 8kHz in both ears.ResultsAlthough aspirin was well-tolerated, it did not protect protect hearing in patients receiving cisplatin [mean difference 9.38 60% CI (-1.45, 20.22) one sided p-value of 0.233 in favour of placebo 71% (56/79) patients demonstrated hearing loss following cisplatin administration (median loss of 35db IQR 0-90 range -85 to 180, mean 42.4 SD 56.3). The extent of hearing-loss was not associated with cisplatin dose, age or hearing loss at presentation. Patients undergoing treatment for head and neck malignancy experienced the biggest hearing loss.ConclusionAspirin was well-tolerated but did not protect hearing suggesting that cisplatin and gentamicin may have distinct ototoxic mechanisms or that cisplatin is more ototoxic, requiring larger protective doses. Cisplatin induced ototoxicity results in significant morbidity and further research is required to prevent it.<br/

COAST (Cisplatin Ototoxicity Attenuated by Aspirin Trial): A phase II double blind, randomised controlled trial to establish if aspirin reduces cisplatin induced hearing loss

BackgroundCisplatin has the highest ototoxic potential of platinum containing drugs leaving 50% of patients with a permanent and irreversible hearing-loss, and associated reduction in quality of life. There are no preventative treatment strategies to minimise this common side-effect. Both cisplatin and gentamicin are thought to damage hearing through a common mechanism involving reactive oxygen species in the inner ear. Aspirin has been shown to minimise gentamicin induced ototoxicity. We tested the hypothesis that aspirin could reduce ototoxicity from cisplatin based chemotherapy.Method94 patients were recruited into a phase II, double blind, placebo controlled, 2-arm trial, and randomised 1:1 from multiple tumour-sites (head and neck, thoracic, bladder, germ cell) in the U.K. Patients underwent pure tone audiometry (PTA) before, and following their final cisplatin dose at 7 and 90 days. Patients received aspirin 975mg tid and omeprazole 20 mg od, or placebos; from the day before, to 2 days after their cisplatin dose, for each cisplatin cycle. The primary endpoint was total hearing difference, comparing pre- and post-cisplatin PTA at 6 and 8kHz in both ears.ResultsAlthough aspirin was well-tolerated, it did not protect protect hearing in patients receiving cisplatin [mean difference 9.38 60% CI (-1.45, 20.22) one sided p-value of 0.233 in favour of placebo 71% (56/79) patients demonstrated hearing loss following cisplatin administration (median loss of 35db IQR 0-90 range -85 to 180, mean 42.4 SD 56.3). The extent of hearing-loss was not associated with cisplatin dose, age or hearing loss at presentation. Patients undergoing treatment for head and neck malignancy experienced the biggest hearing loss.ConclusionAspirin was well-tolerated but did not protect hearing suggesting that cisplatin and gentamicin may have distinct ototoxic mechanisms or that cisplatin is more ototoxic, requiring larger protective doses. Cisplatin induced ototoxicity results in significant morbidity and further research is required to prevent it.<br/

Structural and non-coding variants increase the diagnostic yield of clinical whole genome sequencing for rare diseases

Background: Whole genome sequencing is increasingly being used for the diagnosis of patients with rare diseases. However, the diagnostic yields of many studies, particularly those conducted in a healthcare setting, are often disappointingly low, at 25–30%. This is in part because although entire genomes are sequenced, analysis is often confined to in silico gene panels or coding regions of the genome. Methods: We undertook WGS on a cohort of 122 unrelated rare disease patients and their relatives (300 genomes) who had been pre-screened by gene panels or arrays. Patients were recruited from a broad spectrum of clinical specialties. We applied a bioinformatics pipeline that would allow comprehensive analysis of all variant types. We combined established bioinformatics tools for phenotypic and genomic analysis with our novel algorithms (SVRare, ALTSPLICE and GREEN-DB) to detect and annotate structural, splice site and non-coding variants. Results: Our diagnostic yield was 43/122 cases (35%), although 47/122 cases (39%) were considered solved when considering novel candidate genes with supporting functional data into account. Structural, splice site and deep intronic variants contributed to 20/47 (43%) of our solved cases. Five genes that are novel, or were novel at the time of discovery, were identified, whilst a further three genes are putative novel disease genes with evidence of causality. We identified variants of uncertain significance in a further fourteen candidate genes. The phenotypic spectrum associated with RMND1 was expanded to include polymicrogyria. Two patients with secondary findings in FBN1 and KCNQ1 were confirmed to have previously unidentified Marfan and long QT syndromes, respectively, and were referred for further clinical interventions. Clinical diagnoses were changed in six patients and treatment adjustments made for eight individuals, which for five patients was considered life-saving. Conclusions: Genome sequencing is increasingly being considered as a first-line genetic test in routine clinical settings and can make a substantial contribution to rapidly identifying a causal aetiology for many patients, shortening their diagnostic odyssey. We have demonstrated that structural, splice site and intronic variants make a significant contribution to diagnostic yield and that comprehensive analysis of the entire genome is essential to maximise the value of clinical genome sequencing.</p