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A generalised, simple and efficient synthesis of N-acetyl-5-bromo-4-chloroindoxyl and related analogues required for the synthesis of indigogenic substrates to probe for biological activities is reported. The method is both synthetically and operationally simple and represents a significant improvement on existing methods.
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Corantes/síntese química , Índigo Carmim/síntese química , Indóis/síntese química , Benzoatos/química , Índigo Carmim/análogos & derivados , Indóis/químicaRESUMO
Herbicides are an essential tool not only in weed management, but also in conservation tillage approaches to cropping. The first commercial herbicides were released in the 1940s and hundreds more since then, although genetic resistance to them is an issue. Here, we review the experimental and estimated physicochemical properties of 334 successful herbicidal compounds and make available a dynamic electronic database containing detailed analyses of the main chemical properties for herbicides and which adopts the Simplified Molecular-Input Line-Entry System (SMILES) for describing the structure of chemical molecules. This fully available resource allows for the rapid comparison of potential new herbicidal compounds to the chemical properties of known herbicides.
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Fenômenos Químicos , Bases de Dados de Produtos Farmacêuticos , Herbicidas/química , Herbicidas/farmacologia , Plantas Daninhas/efeitos dos fármacos , Animais , Avaliação Pré-Clínica de Medicamentos , Herbicidas/análise , HumanosRESUMO
C-N bond forming reactions are important in organic chemistry. A thin film microfluidic vortex fluidic device (VFD) operating under confined mode affords N-aryl compounds from 2-chloropyrazine and the corresponding amine, without the need for a transition metal catalyst.
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Aminas/química , Fenômenos Químicos , Aminas/síntese química , Catálise , Microfluídica , Morfolinas/química , Paládio/química , Pirazinas/químicaRESUMO
Treatment of Parkinson's disease with dopaminergic agents, such as l-DOPA, is frequently compromised by disabling side effects, particularly dyskinesia and a shortening in duration of antiparkinsonian action. Studies in animal models and anecdotal evidence from a patient with Parkinson's disease show that the illicit drug ecstasy (MDMA) can alleviate these side effects, though with many drawbacks (e.g., psychoactivity). MDMA itself thus has little therapeutic potential. On the basis of known structure-psychoactivity relationships, we designed a series of α-substituted MDMA analogues, one of which, bearing an α-cyclopropyl substituent (UWA-101), enhanced the quality of l-DOPA actions in animal models. Indeed, UWA-101 was more effective than MDMA. Unlike MDMA, UWA-101 did not reduce viability of serotonergic cells, exhibit psychoactive properties, or reduce food intake, and did not substitute for MDMA in drug discrimination assays. UWA-101 displayed a unique receptor/transporter binding profile relative to MDMA, with a >5-fold decrease in affinity for NET and 5-HT(2A) receptors and a 10-fold increase in affinity for DAT. Furthermore, in a functional reuptake assay, UWA-101 inhibited both 5-HT and dopamine reuptake, while having no effect on the reuptake of noradrenaline. UWA-101 is the first selective DAT/SERT inhibitor described with comparable affinities for these two sites. These data identify a new class of therapeutic in Parkinson's disease and highlight the potential benefits of studying illicit drugs that in themselves would never be considered safe for long-term therapy.
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Antiparkinsonianos/uso terapêutico , Benzodioxóis/uso terapêutico , Levodopa/uso terapêutico , Metilaminas/uso terapêutico , N-Metil-3,4-Metilenodioxianfetamina/análogos & derivados , Doença de Parkinson/tratamento farmacológico , Animais , Linhagem Celular Tumoral , Sinergismo Farmacológico , Masculino , N-Metil-3,4-Metilenodioxianfetamina/uso terapêutico , Ratos , Ratos Sprague-DawleyRESUMO
While 3,4-methylenedioxymethamphetamine (MDMA/'ecstasy') is cytostatic towards lymphoma cells in vitro, the concentrations required militate against its translation directly to a therapeutic in vivo. The possibility of 'redesigning the designer drug', separating desired anti-lymphoma activity from unwanted psychoactivity and neurotoxicity, was therefore mooted. From an initial analysis of MDMA analogues synthesized with a modified α-substituent, it was found that incorporating a phenyl group increased potency against sensitive, Bcl-2-deplete, Burkitt's lymphoma (BL) cells 10-fold relative to MDMA. From this lead, related analogs were synthesized with the 'best' compounds (containing 1- and 2-naphthyl and para-biphenyl substituents) some 100-fold more potent than MDMA versus the BL target. When assessed against derived lines from a diversity of B-cell tumors MDMA analogues were seen to impact the broad spectrum of malignancy. Expressing a BCL2 transgene in BL cells afforded only scant protection against the analogues and across the malignancies no significant correlation between constitutive Bcl-2 levels and sensitivity to compounds was observed. Bcl-2-deplete cells displayed hallmarks of apoptotic death in response to the analogues while BCL2 overexpressing equivalents died in a caspase-3-independent manner. Despite lymphoma cells expressing monoamine transporters, their pharmacological blockade failed to reverse the anti-lymphoma actions of the analogues studied. Neither did reactive oxygen species account for ensuing cell death. Enhanced cytotoxic performance did however track with predicted lipophilicity amongst the designed compounds. In conclusion, MDMA analogues have been discovered with enhanced cytotoxic efficacy against lymphoma subtypes amongst which high-level Bcl-2--often a barrier to drug performance for this indication--fails to protect.
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Linfoma de Burkitt/tratamento farmacológico , Linfoma de Burkitt/patologia , Desenho de Fármacos , N-Metil-3,4-Metilenodioxianfetamina/uso terapêutico , Transdução de Sinais , Linfócitos B/patologia , Linfoma de Burkitt/metabolismo , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Humanos , N-Metil-3,4-Metilenodioxianfetamina/análogos & derivados , N-Metil-3,4-Metilenodioxianfetamina/química , N-Metil-3,4-Metilenodioxianfetamina/farmacologia , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Transdução de Sinais/efeitos dos fármacosRESUMO
OBJECTIVE: The purpose of this study was to investigate rigid ankle-foot orthosis (AFO) prescription practices for adult men among UK orthotists. DESIGN: A cross-sectional study using a survey was distributed online to UK orthotists by the British Association of Prosthetists and Orthotists to its members and through social media and orthotic networks. The survey was completed between November 1, 2020, and November 29, 2020. MAIN OUTCOME MEASURES: Descriptive statistics of survey results include information related to the material used, the thickness of the material, positive cast rectification, AFO reinforcement, footplate design, padding, strapping system, and height of AFO. RESULTS: One hundred participants completed the survey, which equates to a response rate of 30.5% of the British Association of Prosthetists and Orthotists members targeted. A clear consensus emerged on the design of a bespoke rigid AFO for the hypothetical patient in this study, which is detailed as follows: 1) 4.5 mm copolymer polypropylene, 2) no additional reinforcement, 3) full-length footplate with mediolateral trimlines terminating behind the metatarsal heads, 4) 3-point correction with parallel sides, 5) padded VELCRO straps with D-rings at the calf and heel, 6) no forefoot or other additional strapping, 7) 3-mm PORON (international Ltd) padding at the malleoli, and 8) AFO height that finishes 2 cm below the fibular head. CONCLUSIONS: This study has highlighted a consensus on AFO prescription/design among UK orthotists surveyed, based on the hypothetical patient described in this study.
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Órtoses do Pé , Adulto , Masculino , Humanos , Tornozelo , Estudos Transversais , Prescrições , Reino Unido , Fenômenos Biomecânicos , Desenho de EquipamentoRESUMO
OBJECTIVES: To assess whether a commercially available CE-IVD, ELISA-based surrogate neutralisation assay (cPass, Genscript) provides a genuine measure of SARS-CoV-2 neutralisation by human sera, and further to establish whether measuring responses against the RBD of S was a diagnostically useful proxy for responses against the whole S protein. METHODS: Serum samples from 30 patients were assayed for anti-NP responses, for 'neutralisation' by the surrogate neutralisation assay and for neutralisation by SARS-CoV-2 S pseudotyped virus assays utilising two target cell lines. Correlation between assays was measured using linear regression. RESULTS: The responses observed within the surrogate neutralisation assay demonstrated an extremely strong, highly significant positive correlation with those observed in both pseudotyped virus assays. CONCLUSIONS: The tested ELISA-based surrogate assay provides an immunologically useful measure of functional immune responses in a much quicker and highly automatable fashion. It also reinforces that detection of anti-RBD neutralising antibodies alone is a powerful measure of the capacity to neutralise viral infection.
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COVID-19 , SARS-CoV-2 , Anticorpos Neutralizantes , Anticorpos Antivirais , Ensaio de Imunoadsorção Enzimática , HumanosRESUMO
PURPOSE: The objective for this study was to determine a means of using non-cognitive variables to target undergraduate efforts towards likely candidates for specific residency choice. APPROACH AND MATERIALS: Over a three year spread, a volunteer cohort of freshmen medical students was assessed for the non-cognitive variable of personality preference using the Myers-Briggs Type Indicator (MBTI) to determine its accuracy in predicting the choice of specialty training. RESULTS: Of the 98 participants, 13 ultimately chose a residency in Obstetrics and Gynecology (OB/GYN). Four had scores which classified them with the personality preference ESFP (Extroverted, Sensing, Feeling, Perceiving), representing a ratio of 3.5 to 1 when compared to students expressing other personality preferences among the total sample of subjects. CONCLUSION: These preliminary findings indicated that classification of personality preference among freshmen medical students is a statistically reliable means of predicting future residency choices.
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Escolha da Profissão , Internato e Residência , Personalidade , Estudantes de Medicina/psicologia , Adulto , Feminino , Ginecologia/educação , Humanos , Masculino , Obstetrícia/educação , Inventário de PersonalidadeRESUMO
Early oesophageal adenocarcinoma (OA) and pre-neoplastic dysplasia may be treated with endoscopic resection and ablative techniques such as photodynamic therapy (PDT). Though effective, discrete areas of disease may be missed leading to recurrence. PDT further suffers from the side effects of off-target photosensitivity. A tumour specific and light targeted therapeutic agent with optimised pharmacokinetics could be used to destroy residual cancerous cells left behind after resection. A small molecule antibody-photosensitizer conjugate was developed targeting human epidermal growth factor receptor 2 (HER2). This was tested in an in vivo mouse model of human OA using a xenograft flank model with clinically relevant low level HER2 expression and heterogeneity. In vitro we demonstrate selective binding of the conjugate to tumour versus normal tissue. Light dependent cytotoxicity of the phototherapy agent in vitro was observed. In an in vivo OA mouse xenograft model the phototherapy agent had desirable pharmacokinetic properties for tumour uptake and blood clearance time. PDT treatment caused tumour growth arrest in all the tumours despite the tumours having a clinically defined low/negative HER2 expression level. This new phototherapy agent shows therapeutic potential for treatment of both HER2 positive and borderline/negative OA.
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BACKGROUND: Mucin glycoprotein 1 (MUC1) is a glycosylated transmembrane protein on epithelial cells. We investigate MUC1 as a therapeutic target in Barrett's epithelium (BE) and esophageal adenocarcinoma (EA) and provide proof of concept for a light based therapy targeting MUC1. RESULTS: MUC1 was present in 21% and 30% of significantly enriched pathways comparing BE and EA to squamous epithelium respectively. MUC1 gene expression was x2.3 and x2.2 higher in BE (p=<0.001) and EA (p=0.03). MUC1 immunohistochemical expression increased during progression to EA and followed tumor invasion. HuHMFG1 based photosensitive antibody drug conjugates (ADC) showed cell internalization, MUC1 selective and light-dependent cytotoxicity (p=0.0006) and superior toxicity over photosensitizer alone (p=0.0022). METHODS: Gene set enrichment analysis (GSEA) evaluated pathways during BE and EA development and quantified MUC1 gene expression. Immunohistochemistry and flow cytometry evaluated the anti-MUC1 antibody HuHMFG1 in esophageal cells of varying pathological grade. Confocal microscopy examined HuHMFG1 internalization and HuHMFG1 ADCs were created to deliver a MUC1 targeted phototoxic payload. CONCLUSIONS: MUC1 is a promising target in EA. Molecular and light based targeting of MUC1 with a photosensitive ADC is effective in vitro and after development may enable treatment of locoregional tumors endoscopically.
Assuntos
Adenocarcinoma/genética , Antineoplásicos Imunológicos/farmacologia , Neoplasias Esofágicas/genética , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Imunoconjugados/farmacologia , Luz , Mucina-1/genética , Adenocarcinoma/tratamento farmacológico , Adenocarcinoma/metabolismo , Adenocarcinoma/patologia , Biomarcadores , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Biologia Computacional , Progressão da Doença , Neoplasias Esofágicas/tratamento farmacológico , Neoplasias Esofágicas/metabolismo , Neoplasias Esofágicas/patologia , Perfilação da Expressão Gênica , Glicoproteínas/genética , Glicoproteínas/metabolismo , Humanos , Imuno-Histoquímica , Mucina-1/metabolismo , Gradação de Tumores , Metástase NeoplásicaRESUMO
Purpose Retrospective studies indicate that expression of excision repair cross complementing group 1 (ERCC1) protein is associated with platinum resistance and survival in non-small-cell lung cancer (NSCLC). We conducted the first randomized trial, to our knowledge, to evaluate ERCC1 prospectively and to assess the superiority of nonplatinum therapy over platinum doublet therapy for ERCC1-positive NSCLC as well as noninferiority for ERCC1-negative NSCLC. Patients and Methods This trial had a marker-by-treatment interaction phase III design, with ERCC1 (8F1 antibody) status as a randomization stratification factor. Chemonaïve patients with NSCLC (stage IIIB and IV) were eligible. Patients with squamous histology were randomly assigned to cisplatin and gemcitabine or paclitaxel and gemcitabine; nonsquamous patients received cisplatin and pemetrexed or paclitaxel and pemetrexed. Primary end point was overall survival (OS). We also evaluated an antibody specific for XPF (clone 3F2). The target hazard ratio (HR) for patients with ERCC1-positive NSCLC was ≤ 0.78. Results Of patients, 648 were recruited (177 squamous, 471 nonsquamous). ERCC1-positive rates were 54.5% and 76.7% in nonsquamous and squamous patients, respectively, and the corresponding XPF-positive rates were 70.5% and 68.5%. Accrual stopped early in 2012 for squamous patients because OS for nonplatinum therapy was inferior to platinum therapy (median OS, 7.6 months [paclitaxel and gemcitabine] v 10.7 months [cisplatin and gemcitabine]; HR, 1.46; P = .02). Accrual for nonsquamous patients halted in 2013. Median OS was 8.0 (paclitaxel and pemetrexed) versus 9.6 (cisplatin and pemetrexed) months for ERCC1-positive patients (HR, 1.11; 95% CI, 0.85 to 1.44), and 10.3 (paclitaxel and pemetrexed) versus 11.6 (cisplatin and pemetrexed) months for ERCC1-negative patients (HR, 0.99; 95% CI, 0.73 to 1.33; interaction P = .64). OS HR was 1.09 (95% CI, 0.83 to 1.44) for XPF-positive patients, and 1.39 (95% CI, 0.90 to 2.15) for XPF-negative patients (interaction P = .35). Neither ERCC1 nor XPF were prognostic: among nonsquamous patients, OS HRs for positive versus negative were ERCC1, 1.11 ( P = .32), and XPF, 1.08 ( P = .55). Conclusion Superior outcomes were observed for patients with squamous histology who received platinum therapy compared with nonplatinum chemotherapy; however, selecting chemotherapy by using commercially available ERCC1 or XPF antibodies did not confer any extra survival benefit.
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Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Biomarcadores Tumorais/metabolismo , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Proteínas de Ligação a DNA/metabolismo , Endonucleases/metabolismo , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores Tumorais/imunologia , Carcinoma Pulmonar de Células não Pequenas/imunologia , Proteínas de Ligação a DNA/imunologia , Intervalo Livre de Doença , Feminino , Humanos , Neoplasias Pulmonares/imunologia , Masculino , Adesão à Medicação , Pessoa de Meia-Idade , Compostos Organoplatínicos/administração & dosagem , Prognóstico , Taxa de SobrevidaRESUMO
We demonstrate the metabolism of ergosterol by cytochrome P450scc in either a reconstituted system or isolated adrenal mitochondria. The major reaction product was identified as 17alpha,24-dihydroxyergosterol. Purified P450scc also generated hydroxyergosterol as a minor product, which is probably an intermediate in the synthesis of 17alpha,24-dihydroxyergosterol. In contrast to cholesterol and 7-dehydrocholesterol, cleavage of the ergosterol side chain was not observed. NMR analysis clearly located one hydroxyl group to C24, with evidence that the second hydroxyl group is at C17. 17alpha,24-Dihydroxyergosterol inhibited cell proliferation of HaCaT keratinocytes and melanoma cells. Thus, in comparison with cholesterol and 7-dehydrocholesterol, the 24-methyl group and the C22-C23 double bond of ergosterol prevent side chain cleavage by P450scc and change the enzyme's hydroxylase activity from C22 and C20, to C24 and C17, generating bioactive product.
Assuntos
Enzima de Clivagem da Cadeia Lateral do Colesterol/metabolismo , Ergosterol/análogos & derivados , Ergosterol/metabolismo , Glândulas Suprarrenais , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Ergosterol/farmacologia , Humanos , Mitocôndrias/metabolismo , Ressonância Magnética Nuclear Biomolecular , Fitosteróis/metabolismo , Relação Estrutura-AtividadeRESUMO
A microfluidic vortex fluidic device (VFD) operating in either confined or continuous mode is effective in high yielding photoredox reactions involving Rose Bengal, with short reaction times. This processing can be translated to multi-components reactions, also with significantly reduced processing times relative to batch processing and channel microfluidic processing, with comparable or improved yields.
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AIMS: Mutation detection accuracy has been described extensively; however, it is surprising that pre-PCR processing of formalin-fixed paraffin-embedded (FFPE) samples has not been systematically assessed in clinical context. We designed a RING trial to (i) investigate pre-PCR variability, (ii) correlate pre-PCR variation with EGFR/BRAF mutation testing accuracy and (iii) investigate causes for observed variation. METHODS: 13 molecular pathology laboratories were recruited. 104 blinded FFPE curls including engineered FFPE curls, cell-negative FFPE curls and control FFPE tissue samples were distributed to participants for pre-PCR processing and mutation detection. Follow-up analysis was performed to assess sample purity, DNA integrity and DNA quantitation. RESULTS: Rate of mutation detection failure was 11.9%. Of these failures, 80% were attributed to pre-PCR error. Significant differences in DNA yields across all samples were seen using analysis of variance (p<0.0001), and yield variation from engineered samples was not significant (p=0.3782). Two laboratories failed DNA extraction from samples that may be attributed to operator error. DNA extraction protocols themselves were not found to contribute significant variation. 10/13 labs reported yields averaging 235.8 ng (95% CI 90.7 to 380.9) from cell-negative samples, which was attributed to issues with spectrophotometry. DNA measurements using Qubit Fluorometry demonstrated a median fivefold overestimation of DNA quantity by Nanodrop Spectrophotometry. DNA integrity and PCR inhibition were factors not found to contribute significant variation. CONCLUSIONS: In this study, we provide evidence demonstrating that variation in pre-PCR steps is prevalent and may detrimentally affect the patient's ability to receive critical therapy. We provide recommendations for preanalytical workflow optimisation that may reduce errors in down-stream sequencing and for next-generation sequencing library generation.
Assuntos
Análise Mutacional de DNA/normas , Receptores ErbB/genética , Fixadores/normas , Formaldeído/normas , Ensaio de Proficiência Laboratorial , Mutação , Inclusão em Parafina/normas , Reação em Cadeia da Polimerase/normas , Proteínas Proto-Oncogênicas B-raf/genética , Fixação de Tecidos/normas , Linhagem Celular Tumoral , Análise Mutacional de DNA/métodos , DNA de Neoplasias/genética , DNA de Neoplasias/isolamento & purificação , Erros de Diagnóstico/prevenção & controle , Fluorometria/normas , Humanos , Variações Dependentes do Observador , Valor Preditivo dos Testes , Reprodutibilidade dos Testes , Espectrofotometria/normas , Fixação de Tecidos/métodos , Transfecção , Reino Unido , Estados Unidos , Fluxo de TrabalhoRESUMO
BACKGROUND: The wearing-OFF phenomenon is a common motor complication of chronic L-3,4-dihydroxyphenylalanine (L-DOPA) therapy for Parkinson's disease. We recently described the discovery of UWA-101, a dual serotonin (SERT) and dopamine (DAT) transporter inhibitor, which increases the duration of "good quality" ON-time provided by L-DOPA in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-lesioned primate. Here, we further characterise the effects of UWA-101 on this extension of ON-time in terms of L-DOPA-induced side-effects in the MPTP-lesioned common marmoset. METHODS: Marmosets were rendered parkinsonian by MPTP injection and "primed" by repeated L-DOPA administration, to exhibit dyskinesia and psychosis-like behaviours. Animals were then administered acute challenges of L-DOPA in combination with UWA-101 (1, 3, 6 and 10 mg/kg) or vehicle. RESULTS: In combination with L-DOPA, UWA-101 (3, 6 and 10 mg/kg) significantly increased duration of ON-time (by 28%, 28%, and 33%, respectively; all P<0.05). UWA-101 (10 mg/kg) significantly extended duration of ON-time without disabling dyskinesia (by 62%, P<0.01). UWA-101 did not exacerbate the severity of dyskinesia (P>0.05). However, at the highest doses (6 and 10 mg/kg), UWA-101 increased the severity of psychosis-like behaviours (P<0.05). CONCLUSIONS: Our results demonstrate that dual SERT/ DAT inhibitors can effectively enhance L-DOPA anti-parkinsonian action, without exacerbating dyskinesia and, as such, represent a promising new therapeutic class for wearing-OFF. However, at higher doses, dual SERT/ DAT inhibitors may exacerbate dopaminergic psychosis.
Assuntos
1-Metil-4-Fenil-1,2,3,6-Tetra-Hidropiridina/farmacologia , Benzodioxóis/farmacologia , Monoaminas Biogênicas/metabolismo , Metilaminas/farmacologia , Atividade Motora/efeitos dos fármacos , Inibidores da Captação de Neurotransmissores/farmacologia , Animais , Callithrix , Relação Dose-Resposta a Droga , Feminino , Levodopa/efeitos adversos , Transtornos dos Movimentos/tratamento farmacológico , Transtornos dos Movimentos/etiologia , Inibidores da Captação de Neurotransmissores/uso terapêuticoRESUMO
International and national guidelines highlight the importance of accuracy, reproducibility, and quality control of in situ hybridization (ISH) methods for testing breast carcinomas. However, few guidelines cover the reporting of ISH cases with "unusual" signal patterns, including, eg, heterogeneity and loss of chromosome enumeration probe or gene signals. These cases are, in fact, relatively frequent, and there is a need for developing evidence- or consensus-based reporting guidelines to ensure consistency of treatment. Following an audit of cases from a single center (including >1,700 cases) we show that approximately 10% of ISH results reflect unusual signal patterns. We illustrate the most common of these patterns and provide reporting guidelines for diagnosticians and recommendations for future research. Our goal is to ensure that in the future such "rogues" are reported in a consistent manner that, ultimately, will be supported by molecular and biochemical evidence.
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Neoplasias da Mama/diagnóstico , Amplificação de Genes , Receptor ErbB-2/genética , Neoplasias da Mama/genética , Neoplasias da Mama/metabolismo , Feminino , Humanos , Hibridização in Situ Fluorescente , Controle de Qualidade , Receptor ErbB-2/metabolismo , Reprodutibilidade dos TestesRESUMO
Synthesis of iminosugar-based affinity-based proteomics probes for use in probing exo-α-glycosidase activity.
Assuntos
Marcadores de Afinidade/metabolismo , Glicosídeo Hidrolases/metabolismo , 1-Desoxinojirimicina/farmacologia , Marcadores de Afinidade/síntese química , Arabidopsis , Reagentes de Ligações Cruzadas/química , Inibidores Enzimáticos/farmacologia , Escherichia coli K12 , Glicosídeo Hidrolases/antagonistas & inibidores , Glicosídeo Hidrolases/química , Processos Fotoquímicos , Proteômica/métodos , Especificidade por Substrato , Raios Ultravioleta , LevedurasRESUMO
These guidelines supplement existing guidelines on HER2 testing by immunohistochemistry and in-situ hybridisation(ISH) methods in the UK. They provide a specific focus on aspects of guidance relevant to HER2 ISH testing methods, both fluorescent and chromogenic. They are formulated to give advice on methodology, interpretation and quality control for ISH-based testing of HER2 status in common tumour types, including both breast and gastric tumours. The aim is to ensure that all ISH-based testing is accurate, reliable and timely.
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Neoplasias da Mama/diagnóstico , Hibridização In Situ/métodos , Receptor ErbB-2/metabolismo , Neoplasias Gástricas/diagnóstico , Neoplasias da Mama/genética , Feminino , Genes erbB-2 , Humanos , Hibridização In Situ/normas , Capacitação em Serviço , Corpo Clínico/educação , Técnicas de Sonda Molecular , Garantia da Qualidade dos Cuidados de Saúde , Controle de Qualidade , Projetos de Pesquisa , Manejo de Espécimes , Neoplasias Gástricas/genéticaAssuntos
Biomarcadores Tumorais/análise , Carcinoma Ductal de Mama/química , Carcinoma Intraductal não Infiltrante/química , Proteínas Nucleares/análise , Fatores de Transcrição/análise , Neoplasias de Mama Triplo Negativas/química , Idoso , Biomarcadores Tumorais/genética , Biópsia , Carcinoma Ductal de Mama/genética , Carcinoma Ductal de Mama/patologia , Carcinoma Ductal de Mama/terapia , Carcinoma Intraductal não Infiltrante/genética , Carcinoma Intraductal não Infiltrante/patologia , Carcinoma Intraductal não Infiltrante/terapia , Diagnóstico Diferencial , Erros de Diagnóstico/prevenção & controle , Feminino , Humanos , Imuno-Histoquímica , Gradação de Tumores , Valor Preditivo dos Testes , Fator Nuclear 1 de Tireoide , Neoplasias de Mama Triplo Negativas/genética , Neoplasias de Mama Triplo Negativas/patologia , Neoplasias de Mama Triplo Negativas/terapiaRESUMO
An improved medium scale synthesis of 3-[(2-methyl-1,3-thiazol-4-yl)ethynyl]pyridine (MTEP), a selective and potent metabotropic glutamate subtype 5 (mGlu5) antagonist, has allowed thorough characterisation of the crystal structures of the free base and the previously unreported hydrochloride (MTEP.HCl). Hirshfeld surface analysis has revealed that molecules in crystalline MTEP are weakly polar, and aggregate through nonclassical C--H...N hydrogen bonds. A strong ionic N--H(+)...Cl(-) hydrogen bond dominates the crystal packing in MTEP.HCl. Despite significant differences in the crystal packing, the molecular structures of MTEP and MTEP.HCl are very similar. The acid dissociation constants for MTEP were investigated using (1)H NMR spectroscopy. The second acid dissociation constant (pK(a2)), associated with the pyridine nitrogen, was determined to be 3.40 +/- 0.01, whilst pK(a1), associated with the thiazole nitrogen, was estimated to be 0.2. The low pK(a) values make it unlikely that MTEP is protonated in its biologically active form.