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BACKGROUND: Brainstem encephalitis is a serious complication of hand foot and mouth disease (HFMD) in children. Autonomic nervous system (ANS) dysregulation and hypertension may occur, sometimes progressing to cardiopulmonary failure and death. Vietnamese national guidelines recommend use of milrinone if ANS dysregulation with Stage 2 hypertension develops. We wished to investigate whether magnesium sulfate (MgSO4) improved outcomes in children with HFMD if used earlier in the evolution of the ANS dysregulation (Stage 1 hypertension). METHODS: During a regional epidemic we conducted a randomized, double-blind, placebo-controlled trial of MgSO4 in children with HFMD, ANS dysregulation and Stage 1 hypertension, at the Hospital for Tropical Diseases in Ho Chi Minh city. Study participants received an infusion of MgSO4 or matched placebo for 72 h. We also reviewed data from non-trial HFMD patients in whom milrinone failed to control hypertension, some of whom received MgSO4 as second line therapy. The primary outcome for both analyses was a composite of disease progression within 72 h - addition of milrinone (trial participants only), need for ventilation, shock, or death. RESULTS: Between June 2014 and September 2016, 14 and 12 participants received MgSO4 or placebo respectively, before the trial was stopped due to futility. Among 45 non-trial cases with poorly controlled hypertension despite high-dose milrinone, 33 received MgSO4 while 12 did not. There were no statistically significant differences in the composite outcome between the MgSO4 and the placebo/control groups in either study (adjusted relative risk (95%CI) of [6/14 (43%) vs. 6/12 (50%)], 0.84 (0.37, 1.92), p = 0.682 in the trial and [1/33 (3%) vs. 2/12 (17%)], 0.16 (0.01, 1.79), p = 0.132 in the observational cohort). The incidence of adverse events was similar between the groups. Potentially toxic magnesium levels occurred very rarely with the infusion regime used. CONCLUSION: Although we could not demonstrate efficacy in these studies, there were no safety signals associated with use of 30-50 mg/kg/hr. MgSO4 in severe HFMD. Intermittent outbreaks of HFMD are likely to continue across the region, and an adequately powered trial is still needed to evaluate use of MgSO4 in controlling hypertension in severe HFMD, potentially involving a higher dose regimen. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT01940250 (Registered 22 AUG 2013). Trial sponsor: University of Oxford.
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Doenças do Sistema Nervoso Autônomo/tratamento farmacológico , Doença de Mão, Pé e Boca/tratamento farmacológico , Sulfato de Magnésio/uso terapêutico , Animais , Sistema Nervoso Autônomo/efeitos dos fármacos , Sistema Nervoso Autônomo/fisiologia , Doenças do Sistema Nervoso Autônomo/etiologia , Criança , Pré-Escolar , Estudos de Coortes , Progressão da Doença , Método Duplo-Cego , Feminino , Doença de Mão, Pé e Boca/complicações , Doença de Mão, Pé e Boca/fisiopatologia , Hemodinâmica/efeitos dos fármacos , Humanos , Lactente , Sulfato de Magnésio/efeitos adversos , Masculino , PlacebosRESUMO
BACKGROUND: Toxicity of airborne particulate matter (PM) is difficult to assess because PM composition is complex and variable due to source contribution and atmospheric transformation. In this study, we used an in vitro toxicoproteomic approach to identify the toxicity mechanisms associated with different subfractions of Ottawa urban dust (EHC-93). METHODS: A549 human lung epithelial cells were exposed to 0, 60, 140 and 200 µg/cm2 doses of EHC-93 (total), its insoluble and soluble fractions for 24 h. Multiple cytotoxicity assays and proteomic analyses were used to assess particle toxicity in the exposed cells. RESULTS: The cytotoxicity data based on cellular ATP, BrdU incorporation and LDH leakage indicated that the insoluble, but not the soluble, fraction is responsible for the toxicity of EHC-93 in A549 cells. Two-dimensional gel electrophoresis results revealed that the expressions of 206 protein spots were significantly altered after particle exposures, where 154 were identified by MALDI-TOF-TOF-MS/MS. The results from cytotoxicity assays and proteomic analyses converged to a similar finding that the effects of the total and insoluble fraction may be alike, but their effects were distinguishable, and their effects were significantly different from the soluble fraction. Furthermore, the toxic potency of EHC-93 total is not equal to the sum of its insoluble and soluble fractions, implying inter-component interactions between insoluble and soluble materials resulting in synergistic or antagonistic cytotoxic effects. Pathway analysis based on the low toxicity dose (60 µg/cm2) indicated that the two subfractions can alter the expression of those proteins involved in pathways including cell death, cell proliferation and inflammatory response in a distinguishable manner. For example, the insoluble and soluble fractions differentially affected the secretion of pro-inflammatory cytokines such as MCP-1 and IL-8 and distinctly altered the expression of those proteins (e.g., TREM1, PDIA3 and ENO1) involved in an inflammatory response pathway in A549 cells. CONCLUSIONS: This study demonstrated the impact of different fractions of urban air particles constituted of various chemical species on different mechanistic pathways and thus on cytotoxicity effects. In vitro toxicoproteomics can be a valuable tool in mapping these differences in air pollutant exposure-related toxicity mechanisms.
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Pulmão/efeitos dos fármacos , Material Particulado/toxicidade , Proteômica/métodos , Solventes/química , Toxicologia/métodos , Água/química , Células A549 , Sobrevivência Celular/efeitos dos fármacos , Quimiocina CCL2/metabolismo , Relação Dose-Resposta a Droga , Eletroforese em Gel Bidimensional , Humanos , Mediadores da Inflamação/metabolismo , Interleucina-8/metabolismo , Pulmão/metabolismo , Pulmão/patologia , Material Particulado/química , Medição de Risco , Transdução de Sinais/efeitos dos fármacos , Solubilidade , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Espectrometria de Massas em Tandem , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
In this study, we used cytotoxicity assays, proteomic and gene expression analyses to examine the difference in response of A549 cells to two silica particles that differ in physical properties, namely cristobalite (CR) and α-quartz (Min-U-Sil 5, MI). Cytotoxicity assays such as lactate dehydrogenase release, 5-bromo-2'-deoxyuridine incorporation and cellular ATP showed that both silica particles could cause cell death, decreased cell proliferation and metabolism in the A549 human lung epithelial cells. While cytotoxicity assays revealed little difference between CR and MI exposures, proteomic and gene expression analyses unveiled both similar and unique molecular changes in A549 cells. For instance, two-dimensional gel electrophoresis data indicated that the expression of proteins in the cell death (e.g., ALDH1A1, HTRA2 and PRDX6) and cell proliferation (e.g., FSCN1, HNRNPAB and PGK1) pathways were significantly different between the two silica particles. Reverse transcription-polymerase chain reaction data provided additional evidence supporting the proteomic findings. Preliminary assessment of the physical differences between CR and MI suggested that the extent of surface interaction between particles and cells could explain some of the observed biological effects. However, the differential dose-response curves for some other genes and proteins suggest that other physical attributes of particulate matter can also contribute to particulate matter-related cellular toxicity. Our results demonstrated that toxicoproteomic and gene expression analyses are sensitive in distinguishing subtle toxicity differences associated with silica particles of varying physical properties compared to traditional cytotoxicity endpoints. Copyright © 2016 Her Majesty the Queen in Right of Canada. Journal of Applied Toxicology published by John Wiley & Sons, Ltd.
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Células Epiteliais/efeitos dos fármacos , Material Particulado/toxicidade , Proteoma/efeitos dos fármacos , Dióxido de Silício/toxicidade , Transcriptoma/efeitos dos fármacos , Células A549 , Técnicas de Cultura de Células , Sobrevivência Celular/efeitos dos fármacos , Eletroforese em Gel Bidimensional , Células Epiteliais/metabolismo , Perfilação da Expressão Gênica/métodos , Humanos , Material Particulado/química , Proteômica/métodos , Quartzo/química , Quartzo/toxicidade , Sensibilidade e Especificidade , Dióxido de Silício/química , Propriedades de SuperfícieRESUMO
BACKGROUND: Industrial sources contribute a significant proportion of anthropogenic particulate matter (PM) emissions, producing particles of varying composition that may differentially impact health. This study investigated the in vitro toxicity of ambient PM collected near industrial sites in relation to particle size and composition. METHODS: Size-fractionated particles (ultrafine, PM0.1-2.5, PM2.5-10, PM>10) were collected in the vicinity of steel, copper, aluminium, and petrochemical industrial sites. Human lung epithelial-like A549 and murine macrophage-like J774A.1 cells were exposed for 24 h to particle suspensions (0, 30, 100, 300 µg/cm2). Particle potency was assessed using cytotoxic (resazurin reduction, lactate dehydrogenase (LDH) release) and inflammatory (cytokine release) assays, and regressed against composition (metals, polycyclic aromatic hydrocarbons (PAHs), endotoxin). RESULTS: Coarse (PM2.5-10, PM>10) particle fractions were composed primarily of iron and aluminium; in contrast, ultrafine and fine (PM0.1-2.5) fractions displayed considerable variability in metal composition (especially water-soluble metals) across collection sites consistent with source contributions. Semi-volatile and PM-associated PAHs were enriched in the fine and coarse fractions collected near metal industry. Cell responses to exposure at equivalent mass concentrations displayed striking differences among sites (SITE x SIZE and SITE x DOSE interactions, p < 0.05), suggesting that particle composition, in addition to size, impacted particle toxicity. While both J774A.1 and A549 cells exhibited clear particle size-dependent effects, site-dependent differences were more pronounced in J774A.1 cells, suggesting greater sensitivity to particle composition. Plotting particle potency according to cytotoxic and inflammatory response grouped particles by size and site, and showed that particles of similar composition tended to cluster together. Cytotoxic effects in J774A.1 cells correlated with metal and PAH content, while inflammatory responses were associated primarily with endotoxin content in coarse particles. CONCLUSIONS: Industrial sources produce particulate emissions with varying chemical composition that differ in their in vitro potency in relation to particle size and the levels of specific constituents.
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Indústrias , Material Particulado/toxicidade , Animais , Linhagem Celular , Citocinas/metabolismo , Humanos , CamundongosRESUMO
Spontaneous isolated superior mesenteric artery dissection (SISMAD) is a rare cause of acute abdominal pain, but could potentially be fatal to patients, and should be recognized soon in the emergency department after excluding other common causes. Computed tomography (CT) is the modality of choice for initial diagnosis and follow-up. Currently there is no evidence-based guidelines for managing SISMAD. A 58-year-old man being suspected of a mesenteric artery dissection was referred to our emergergy department. The patient was monitored, treated conservatively with anticoagulant and discharged after 3 days. Follow-up CT scans at 6 month, 1 year and 1 year and a half post discharge showed a partially occluded false lumen, the diameter of true lumen had increased in size and no signs of bowel ischemia. SISMAD should be considered as part of differential diagnoses when patients in their fifth to seventh decades of life present with acute abdominal pain. Treatment includes conservative management, percutaneous endovascular interventions, or surgery, but most patients can be managed conservatively.
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Introduction: The lack of access to behavioral health care, trends in behavioral health issues, and the impact of social determinants of health underlie the need for behavioral health reform in Kansas. However, stakeholders may affect progress toward behavioral health reform. This study examined stakeholders' attitudes toward behavioral health reform. Methods: The authors analyzed data from a survey administered to elected officials, members of health advocacy groups, state employees, and payers in Kansas. Main outcome measures included attitudes toward the perceived benefit of certain behavioral health and social determinants of health policies and the perceived performance of the primary care and behavioral health care systems in Kansas. Results: Payers perceived legislation to improve insurance coverage for behavioral health issues as less beneficial than state employees and members of health advocacy groups. Elected officials perceived legislation to address various social determinants of health as less beneficial than health advocates. Members of health advocacy groups rated the behavioral health care system more poorly than elected officials did. Conclusions: Preliminary findings reflected both the barriers and facilitators to behavioral health reform in Kansas. However, several limitations undermined the generalizability of these findings. Future studies should consider more representative sample sizes, additional variables in behavioral health and social determinants of health policies, and more comprehensive, validated measures.
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The need for mental health care for pediatric patients outstrips the supply, especially in states, like Kansas, that experience shortages of mental health professionals. Pediatric mental health care access programs, like KSKidsMAP (Kansas Kids Mental health Access Program), increase access to care by building competence and confidence in primary care physicians and clinicians (PCPs) through a statewide integrated system that includes a consultation line. This study is a secondary analysis of KSKidsMAP consultation Line inquiries regarding patients aged 0 to 21 years with mental and behavioral health concerns. The study employs a mixed-method approach with descriptive statistics and thematic analysis of inquiries. Five themes were identified: (1) pharmacotherapy, (2) diagnostic evaluation, (3) community resources, (4) psychotherapy, and (5) other. This study sheds light on PCPs needs and illustrates the importance of Pediatric Mental Health Care Access programs offering interprofessional expertise to consulting PCPs, allowing for expansion of pediatric mental illness care into the primary care setting.
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Saúde Mental , Pneumonia por Pneumocystis , Humanos , Criança , Atenção Primária à Saúde/métodos , Acessibilidade aos Serviços de Saúde , Encaminhamento e Consulta , Pessoal de SaúdeRESUMO
Nationwide challenges with the lack of access to mental health care for youth have prompted efforts to integrate mental health into pediatric primary care. Kansas Kids Mental Health Access Program (KSKidsMAP) was developed to promote mental health workforce development through primary-care practitioners (PCPs) by offering free access to consultations, training, and care coordination. Kansas Kids Mental Health Access Program, a federally funded pediatric mental health care access program (PMHCA), is highly interprofessional in nature, and recommendations reflect the team composition and collaboration efforts. Therefore, a mixed-methods study was conducted to assess the type of recommendations provided to PCPs who requested case consultation services. Seven themes were identified: (1) psychotherapy; (2) diagnostic evaluation; (3) community resources; (4) pharmacotherapy; (5) patient resources and toolkits; (6) education; and (7) other health recommendations. This study highlights the multifaceted approach of KSKidsMAP in addressing PCPs' pediatric mental health concerns.
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Serviços de Saúde Mental , Saúde Mental , Adolescente , Humanos , Criança , Atenção Primária à Saúde/métodos , Encaminhamento e Consulta , Pessoal de SaúdeRESUMO
BACKGROUND: Exposure to different forms of ionizing radiation occurs in diverse occupational, medical, and environmental settings. Improving the accuracy of the estimated health risks associated with exposure is therefore, essential for protecting the public, particularly as it relates to chronic low dose exposures. A key aspect to understanding health risks is precise and accurate modeling of the dose-response relationship. Toward this vision, benchmark dose (BMD) modeling may be a suitable approach for consideration in the radiation field. BMD modeling is already extensively used for chemical hazard assessments and is considered statistically preferable to identifying low and no observed adverse effects levels. BMD modeling involves fitting mathematical models to dose-response data for a relevant biological endpoint and identifying a point of departure (the BMD, or its lower bound). Recent examples in chemical toxicology show that when applied to molecular endpoints (e.g. genotoxic and transcriptional endpoints), BMDs correlate to points of departure for more apical endpoints such as phenotypic changes (e.g. adverse effects) of interest to regulatory decisions. This use of BMD modeling may be valuable to explore in the radiation field, specifically in combination with adverse outcome pathways, and may facilitate better interpretation of relevant in vivo and in vitro dose-response data. To advance this application, a workshop was organized on June 3rd, 2022, in Ottawa, Ontario that brought together BMD experts in chemical toxicology and the radiation scientific community of researchers, regulators, and policy-makers. The workshop's objective was to introduce radiation scientists to BMD modeling and its practical application using case examples from the chemical toxicity field and demonstrate the BMDExpress software using a radiation dataset. Discussions focused on the BMD approach, the importance of experimental design, regulatory applications, its use in supporting the development of adverse outcome pathways, and specific radiation-relevant examples. CONCLUSIONS: Although further deliberations are needed to advance the use of BMD modeling in the radiation field, these initial discussions and partnerships highlight some key steps to guide future undertakings related to new experimental work.
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Benchmarking , Modelos Teóricos , Benchmarking/métodos , Dano ao DNA , Medição de Risco/métodos , Relação Dose-Resposta a DrogaRESUMO
PURPOSE: Benchmark dose (BMD) modeling is a method commonly used in chemical toxicology to identify the point of departure (POD) from a dose-response curve linked to a health-related outcome. Recently, its application in the analysis of transcriptional data for quantitative adverse outcome pathway (AOP) development is being explored. As AOPs are informed by diverse data types, it is important to understand the impact of study parameters such as dose selection, the number of replicates and dose range on BMD outputs for radiation-induced genes and pathways. MATERIALS AND METHODS: Data were selected from the Gene Expression Omnibus (GSE52403) that featured gene expression profiles of peripheral blood samples from C57BL/6 mice 6 hours post-exposure to 137Cs gamma-radiation at 0, 1, 2, 3, 4.5, 6, 8 and 10.5 Gy. The dataset comprised a broad dose range over multiple dose points with consistent dose spacing and multiple biological replicates. This dataset was ideal for systematically transforming across three categories: (1) dose range, (2) dose-spacing and (3) number of controls/replicates. Across these categories, 29 transformed datasets were compared to the original dataset to determine the impact of each transformation on the BMD outputs. RESULTS: Most of the experimental changes did not impact the BMD outputs. The transformed datasets were largely consistent with the original dataset in terms of the number of reproduced genes modeled and absolute BMD values for genes and pathways. Variations in dose selection identified the importance of the absolute value of the lowest and second dose. It was determined that dose selection should include at least two doses <1 Gy and two >5 Gy to achieve meaningful BMD outputs. Changes to the number of biological replicates in the control and non-zero dose groups impacted the overall accuracy and precision of the BMD outputs as well as the ability to fit dose-response models consistent with the original dataset. CONCLUSION: Successful application of transcriptomic BMD modeling for radiation datasets requires considerations of the exposure dose and the number of biological replicates. Most important is the selection of the lowest doses and dose spacing. Reflections on these parameters in experimental design will provide meaningful BMD outputs that could correlate well to apical endpoints of relevance to radiation exposure assessment.
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Benchmarking , Projetos de Pesquisa , Camundongos , Animais , Relação Dose-Resposta a Droga , Medição de Risco/métodos , Camundongos Endogâmicos C57BLRESUMO
PURPOSE: A vast amount of data regarding the effects of radiation stressors on transcriptional changes has been produced over the past few decades. These data have shown remarkable consistency across platforms and experimental design, enabling increased understanding of early molecular effects of radiation exposure. However, the value of transcriptomic data in the context of risk assessment is not clear and represents a gap that is worthy of further consideration. Recently, benchmark dose (BMD) modeling has shown promise in correlating a transcriptional point of departure (POD) to that derived using phenotypic outcomes relevant to human health risk assessment. Although frequently applied in chemical toxicity evaluation, our group has recently demonstrated application within the field of radiation research. This approach allows the possibility to quantitatively compare radiation-induced gene and pathway alterations across various datasets using BMD values and derive meaningful biological effects. However, before BMD modeling can confidently be used, an understanding of the impact of confounding variables on BMD outputs is needed. METHODS: To this end, BMD modeling was applied to a publicly available microarray dataset (Gene Expression Omnibus #GSE23515) that used peripheral blood ex-vivo gamma-irradiated at 0.82 Gy/min, at doses of 0, 0.1, 0.5 or 2 Gy, and assessed 6 hours post-exposure. The dataset comprised six female smokers (F-S), six female nonsmokers (F-NS), six male smokers (M-S), and six male nonsmokers (M-NS). RESULTS: A combined total of 412 genes were fit to models and the BMD distribution was noted to be bi-modal across the four groups. A total of 74, 41, 62 and 62 genes were unique to the F-NS, M-NS, F-S and M-S groups. Sixty-two BMD modeled genes and nine pathways were common across all four groups. There were no differential sensitivity of BMD responses in the robust common genes and pathways. CONCLUSION: For radiation-responsive genes and pathways common across the study groups, the BMD distribution of transcriptional activity was unaltered by sex and smoking status. Although further validation of the data is needed, these initial findings suggest BMD values for radiation relevant genes and pathways are robust and could be explored further in future studies.
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Benchmarking , Radiação Ionizante , Masculino , Humanos , Feminino , Fatores de Confusão Epidemiológicos , Transcriptoma , Medição de RiscoRESUMO
BACKGROUND: Accumulated body of evidence shows that ionizing radiation increases the risk of cataracts. The mechanisms are not clear and the International Commission on Radiological Protection indicates a need for research into understanding the process, particularly at low doses and low dose rates of exposure. PURPOSE: This study was designed to examine protein-level modifications in a human lens epithelial (HLE) cell-line following radiation exposures. MATERIALS AND METHODS: HLE cell-line was subjected to X-irradiation at varied doses (0-5 Gy) and dose-rates (1.62 cGy/min and 38.2 cGy/min). Cells were collected 20 h post-exposure, lysed and proteins were clarified following fractionation by a molecular weight cut-off filtration method. Fractionated cellular proteins were enzymatically digested and subjected to mass spectrometry analysis. RESULTS: Statistically significant radiation dose-related protein changes compared to the control group were identified. Heatmap and hierarchical clustering analysis showed dose-rate dependant responses. Pathway analysis mapped the proteins to biological functions of mitochondrial dysfunction, reactive oxygen species generation, cell death, cancer, organismal injury and amyloidosis. CONCLUSION: Overall findings suggest that ionizing radiation exposure of HLE cells by mediating dose rate-dependant oxidative stress and cell death-related mechanisms, can be relevant to cataractogenesis.
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Células Epiteliais/efeitos da radiação , Cristalino/patologia , Lesões por Radiação/patologia , Linhagem Celular , Relação Dose-Resposta à Radiação , Células Epiteliais/patologia , Humanos , Lesões por Radiação/etiologia , Raios X/efeitos adversosRESUMO
BACKGROUND: Since the COVID-19 pandemic began, there have been concerns related to the preparedness of healthcare workers (HCWs). This study aimed to describe the level of awareness and preparedness of hospital HCWs at the time of the first wave. METHODS: This multinational, multicenter, cross-sectional survey was conducted among hospital HCWs from February to May 2020. We used a hierarchical logistic regression multivariate analysis to adjust the influence of variables based on awareness and preparedness. We then used association rule mining to identify relationships between HCW confidence in handling suspected COVID-19 patients and prior COVID-19 case-management training. RESULTS: We surveyed 24,653 HCWs from 371 hospitals across 57 countries and received 17,302 responses from 70.2% HCWs overall. The median COVID-19 preparedness score was 11.0 (interquartile range [IQR] = 6.0-14.0) and the median awareness score was 29.6 (IQR = 26.6-32.6). HCWs at COVID-19 designated facilities with previous outbreak experience, or HCWs who were trained for dealing with the SARS-CoV-2 outbreak, had significantly higher levels of preparedness and awareness (p<0.001). Association rule mining suggests that nurses and doctors who had a 'great-extent-of-confidence' in handling suspected COVID-19 patients had participated in COVID-19 training courses. Male participants (mean difference = 0.34; 95% CI = 0.22, 0.46; p<0.001) and nurses (mean difference = 0.67; 95% CI = 0.53, 0.81; p<0.001) had higher preparedness scores compared to women participants and doctors. INTERPRETATION: There was an unsurprising high level of awareness and preparedness among HCWs who participated in COVID-19 training courses. However, disparity existed along the lines of gender and type of HCW. It is unknown whether the difference in COVID-19 preparedness that we detected early in the pandemic may have translated into disproportionate SARS-CoV-2 burden of disease by gender or HCW type.
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COVID-19/epidemiologia , Conhecimentos, Atitudes e Prática em Saúde , Recursos Humanos em Hospital , Adulto , COVID-19/prevenção & controle , Estudos Transversais , Educação Médica Continuada/estatística & dados numéricos , Feminino , Humanos , Masculino , Recursos Humanos em Hospital/estatística & dados numéricos , Fatores Socioeconômicos , Inquéritos e QuestionáriosRESUMO
Osteoarthritis (OA) is the most common form of arthritis and the fastest growing cause of chronic disability in the world. Formation of the ternary IL-1ß /IL-1R1/IL-1RAcP protein complex and its downstream signaling has been implicated in osteoarthritis pathology. Current OA therapeutic approaches target either the cytokine IL-1ß or the primary receptor IL-1RI but do not exploit the potential of the secondary receptor IL-1RAcP. Our previous work implicated the Arg286 residue of IL-1RAcP as a key mediator of complex formation. Molecular modeling confirmed Arg286 as a high-energy mediator of the ternary IL-1ß complex architecture and interaction network. Anti-IL-1RAcP monoclonal antibodies (mAb) targeting the Arg286 residue were created and were shown to effectively reduce the influx of inflammatory cells to damaged joints in a mouse model of osteoarthritis. Inhibitory peptides based on the native sequence of IL-1RAcP were prepared and examined for efficacy at disrupting the complex formation. The most potent peptide inhibitor had an IC50 value of 304 pM in a pull-down model of complex formation, and reduced IL-1ß signaling in a cell model by 90% at 2 µM. Overall, therapies that target the Arg286 region surface of IL-1RAcP, and disrupt subsequent interactions with subunits, have the potential to serve as next generation treatments for osteoarthritis.
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Mass spectrometry enhanced by nanotechnology can achieve previously unattainable sensitivity for characterizing urinary pathogen-derived peptides. We utilized mass spectrometry enhanced by affinity hydrogel particles (analytical sensitivity = 2.5 pg/mL) to study tick pathogen-specific proteins shed in the urine of patients with (1) erythema migrans rash and acute symptoms, (2) post treatment Lyme disease syndrome (PTLDS), and (3) clinical suspicion of tick-borne illnesses (TBI). Targeted pathogens were Borrelia, Babesia, Anaplasma, Rickettsia, Ehrlichia, Bartonella, Francisella, Powassan virus, tick-borne encephalitis virus, and Colorado tick fever virus. Specificity was defined by 100% amino acid sequence identity with tick-borne pathogen proteins, evolutionary taxonomic verification for related pathogens, and no identity with human or other organisms. Using a cut off of two pathogen peptides, 9/10 acute Lyme Borreliosis patients resulted positive, while we identified zero false positive in 250 controls. Two or more pathogen peptides were identified in 40% of samples from PTLDS and TBI patients (categories 2 and 3 above, n = 59/148). Collectively, 279 distinct unique tick-borne pathogen derived peptides were identified. The number of pathogen specific peptides was directly correlated with presence or absence of symptoms reported by patients (ordinal regression pseudo-R2 = 0.392, p = 0.010). Enhanced mass spectrometry is a new tool for studying tick-borne pathogen infections.
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Doença de Lyme/microbiologia , Doença de Lyme/urina , Peptídeos/urina , Carrapatos , Adulto , Idoso , Algoritmos , Animais , Babesia microti/metabolismo , Biomarcadores/metabolismo , Borrelia , Eritema Migrans Crônico/microbiologia , Eritema Migrans Crônico/urina , Exantema , Feminino , Humanos , Hidrogéis/química , Infectologia , Masculino , Espectrometria de Massas , Mesocricetus , Pessoa de Meia-Idade , Peptídeos/química , Análise de Regressão , UrináliseRESUMO
PURPOSE: In 2012, the Organization for Economic Cooperation and Development (OECD) formally launched the Adverse Outcome Pathway (AOP) Programme. The AOP framework has the potential for predictive utility in identifying early biological endpoints linked to adverse effects. It uses the weight of correlative evidence to identify a minimal set of measurable key events that link molecular initiating events to an adverse outcome. AOPs have the capability to identify knowledge gaps and priority areas for future research based on relevance to an adverse outcome. In addition, AOPs can identify pathways that are common among multiple stressors, thereby allowing for the possibility of refined risk assessments based on co-exposure considerations. The AOP framework is increasingly being used in chemical and ecological risk assessment; however, its use in the development of radiation-specific pathways has yet to be fully explored. To bring awareness of the AOP framework to the Canadian radiation community, a workshop was held in Canada in June 2018 that brought together radiation experts from Health Canada, the Canadian Nuclear Laboratories, and the Canadian Nuclear Safety Commission. METHODS: The purpose of the workshop was to share knowledge on the AOP framework, specifically (1) to introduce the concept of the AOP framework and its possible utility to Canadian radiation experts; (2) to provide examples on how it has advanced risk assessment; (3) to discuss an illustrative example specific to ionizing radiation; and lastly (4) to identify the broad benefits and challenges of the AOP framework to the radiation community. RESULTS: The participants showed interest in the framework, case examples were described and areas of challenge were identified. Herein, we summarize the outcomes of the workshop. CONCLUSIONS: Overall, participants agreed that by building AOPs in the radiation field, a network of data-sharing initiatives will enhance our interpretation of existing knowledge where current scientific evidence is minimal. They would provide new avenues to understand effects at low-dose and dose-rates and help to quantify the combined effect of multiple stressors on shared mechanistic pathways.
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Rotas de Resultados Adversos , Proteção Radiológica , Humanos , Doses de Radiação , Medição de RiscoRESUMO
We tested the hypothesis that membrane lipid composition influences drug action at membrane proteins by studying local anesthetic action at the nicotinic acetylcholine receptor (nAChR). Infrared difference spectra show that concentrations of tetracaine consistent with binding to the ion channel (<50 microM) stabilize a resting-like state when the nAChR is reconstituted into phosphatidylcholine membranes containing the anionic lipid, phosphatidic acid, but have no effect on the nAChR reconstituted into membranes lacking phosphatidic acid, either in the presence or absence of cholesterol. Concentrations of tetracaine above 200 microM lead to neurotransmitter site binding in all membranes. In the presence of phosphatidic acid, cholesterol, or both, neurotransmitter site binding leads to the formation of quaternary amine-aromatic interactions between tetracaine and binding site tyrosine/tryptophan residues and the stabilization of a desensitized state. One interpretation suggested by lipid partitioning studies is that phosphatidic acid enhances tetracaine action at the channel pore by increasing the partitioning of tetracaine into the lipid bilayer, thereby enhancing access to the transmembrane pore. However, subtle membrane-dependent variations in the vibrations of tyrosine and tryptophan residues, and agonist analog binding studies indicate that the structures of the agonist-bound neurotransmitter sites of the nAChR in membranes lacking both phosphatidic acid and cholesterol differ from the structures of the agonist-desensitized neurotransmitter sites in the presence of both lipids. Lipid action at the nAChR thus involves more than a simple modulation of the equilibrium between resting and desensitized states.
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Colesterol/química , Lipídeos/química , Ácidos Fosfatídicos/química , Fosfatidilcolinas/química , Receptores Nicotínicos/química , Anestésicos Locais/metabolismo , Anestésicos Locais/farmacologia , Animais , Relação Dose-Resposta a Droga , Concentração Inibidora 50 , Cinética , Bicamadas Lipídicas/química , Conformação Proteica , Receptores Nicotínicos/efeitos dos fármacos , Receptores Nicotínicos/isolamento & purificação , Receptores Nicotínicos/metabolismo , Espectroscopia de Infravermelho com Transformada de Fourier , Tetracaína/metabolismo , Tetracaína/farmacologia , TorpedoRESUMO
BACKGROUND/AIM: This study aimed to create a predictive tool for estimating the remaining lifespan of patients after whole-brain irradiation (WBI) for cerebral metastases from bladder cancer. PATIENTS AND METHODS: In 34 of these patients clinical parameters were analyzed for survival including age at start of WBI, gender, Karnofsky score, number of cerebral metastases and involvement of extra-cranial sites of metastasis. RESULTS: Involvement of extra-cranial sites (14%) and Karnofsky score (9%) showed the greatest difference regarding 6-month survival and were considered for the tool. Points were assigned based on the following: no involvement of extra-cranial sites=1 point, involvement of extra-cranial sites=0 points, Karnofsky score ≥70=1 point, Karnofsky score ≤60=0 points. Patients' scores were 0, 1 or 2 points with 6-month survival rates of 13%, 27% and 50%, respectively. CONCLUSION: Based on two clinical parameters, a tool was developed that may help estimate the lifespan of patients irradiated for cerebral metastases from bladder cancer.
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Neoplasias Encefálicas/mortalidade , Neoplasias Encefálicas/secundário , Neoplasias da Bexiga Urinária/patologia , Idoso , Idoso de 80 Anos ou mais , Neoplasias Encefálicas/radioterapia , Irradiação Craniana/métodos , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Doses de Radiação , Resultado do TratamentoRESUMO
BACKGROUND/AIM: To identify predictors of local control and survival after whole-brain irradiation (WBI) for cerebral metastases from cancer of unknown primary (CUP). PATIENTS AND METHODS: In 140 patients receiving WBI alone or following resection, seven factors were investigated including treatment approach, WBI-regimen, age, gender, Eastern Cooperative Oncology Group (ECOG) performance score, number of cerebral lesions and extra-cerebral metastases. RESULTS: On univariate analysis, resection plus WBI and boost (p=0.002), ECOG 0-1 (p<0.001) and a single lesion (p<0.001) were positively associated with local control. On Cox regression, ECOG-score remained significant (p=0.002). On univariate analysis of survival, surgery plus WBI and boost (p=0.009), ECOG 0-1 (p<0.001), a single lesion (p=0.024) and no extra-cerebral metastases (p<0.001) were associated with better outcomes. On Cox regression, ECOG-score (p<0.001) and extra-cerebral lesions (p<0.001) were significant. CONCLUSION: Significant predictors of local control and survival were identified that contribute to treatment personalization and design of prospective trials in patients with cerebral metastases from CUP.
Assuntos
Neoplasias Encefálicas/diagnóstico , Neoplasias Encefálicas/mortalidade , Neoplasias Encefálicas/radioterapia , Neoplasias Primárias Desconhecidas/diagnóstico , Neoplasias Primárias Desconhecidas/mortalidade , Neoplasias Primárias Desconhecidas/radioterapia , Adulto , Idoso , Idoso de 80 Anos ou mais , Neoplasias Encefálicas/secundário , Irradiação Craniana , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Dosagem Radioterapêutica , Estudos Retrospectivos , Análise de Sobrevida , Resultado do TratamentoRESUMO
The likelihood of environmental and health impacts of silicon dioxide nanoparticles (SiNPs) has risen, due to their increased use in products and applications. The biological potency of a set of similarly-sized amorphous SiNPs was investigated in a variety of cells to examine the influence of physico-chemical and biological factors on their toxicity. Cellular LDH and ATP, BrdU incorporation, resazurin reduction and cytokine release were measured in human epithelial A549, human THP-1 and mouse J774A.1 macrophage cells exposed for 24 h to suspensions of 5-15, 10-20 and 12 nm SiNPs and reference particles. The SiNPs were characterized in dry state and in suspension to determine their physico-chemical properties. The dose-response data were simplified into particle potency estimates to facilitate the comparison of multiple endpoints of biological effects in cells. Mouse macrophages were the most sensitive to SiNP exposures. Cytotoxicity of the individual cell lines was correlated while the cytokine responses differed, supported by cell type-specific differences in inflammation-associated pathways. SiNP (12 nm), the most cytotoxic and inflammogenic nanoparticle had the highest surface acidity, dry-state agglomerate size, the lowest trace metal and organics content, the smallest surface area and agglomerate size in suspension. Particle surface acidity appeared to be the most significant determinant of the overall biological activity of this set of nanoparticles. Combined with the nanoparticle characterization, integration of the biological potency estimates enabled a comprehensive determination of the cellular reactivity of the SiNPs. The approach shows promise as a useful tool for first-tier screening of SiNP toxicity.