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1.
Sci Rep ; 11(1): 16051, 2021 08 06.
Artigo em Inglês | MEDLINE | ID: mdl-34362974

RESUMO

With an increasing body of evidence that SARS-CoV-2 is an airborne pathogen, droplet character formed during speech, coughs, and sneezes are important. Larger droplets tend to fall faster and are less prone to drive the airborne transmission pathway. Alternatively, small droplets (aerosols) can remain suspended for long time periods. The small size of SARS-CoV-2 enables it to be encapsulated in these aerosols, thereby increasing the pathogen's ability to be transmitted via airborne paths. Droplet formation during human respiratory events relates to airspeed (speech, cough, sneeze), fluid properties of the saliva/mucus, and the fluid content itself. In this work, we study the fluidic drivers (fluid properties and content) and their influence on factors relating to transmissibility. We explore the relationship between saliva fluid properties and droplet airborne transmission paths. Interestingly, the natural human response appears to potentially work with these drivers to mitigate pathogen transmission. In this work, the saliva is varied using two approaches: (1) modifying the saliva with colloids that increase the viscosity/surface tension, and (2) stimulating the saliva content to increased/decreased levels. Through modern experimental and numerical flow diagnostic methods, the character, content, and exposure to droplets and aerosols are all evaluated. The results indicate that altering the saliva properties can significantly impact the droplet size distribution, the formation of aerosols, the trajectory of the bulk of the droplet plume, and the exposure (or transmissibility) to droplets. High-fidelity numerical methods used and verify that increased droplet size character enhances droplet fallout. In the context of natural saliva response, we find previous studies indicating natural human responses of increased saliva viscosity from stress and reduced saliva content from either stress or illness. These responses both favorably correspond to reduced transmissibility. Such a finding also relates to potential control methods, hence, we compared results to a surgical mask. In general, we find that saliva alteration can produce fewer and larger droplets with less content and aerosols. Such results indicate a novel approach to alter SARS-CoV-2's transmission path and may act as a way to control the COVID-19 pandemic, as well as influenza and the common cold.


Assuntos
COVID-19/transmissão , SARS-CoV-2/isolamento & purificação , Saliva/virologia , Aerossóis/química , Microbiologia do Ar , Coloides/química , Tosse , Humanos , Pandemias , Saliva/química , Espirro , Viscosidade
2.
PLoS One ; 16(8): e0255690, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34351984

RESUMO

Saliva is an attractive specimen type for asymptomatic surveillance of COVID-19 in large populations due to its ease of collection and its demonstrated utility for detecting RNA from SARS-CoV-2. Multiple saliva-based viral detection protocols use a direct-to-RT-qPCR approach that eliminates nucleic acid extraction but can reduce viral RNA detection sensitivity. To improve test sensitivity while maintaining speed, we developed a robotic nucleic acid extraction method for detecting SARS-CoV-2 RNA in saliva samples with high throughput. Using this assay, the Free Asymptomatic Saliva Testing (IGI FAST) research study on the UC Berkeley campus conducted 11,971 tests on supervised self-collected saliva samples and identified rare positive specimens containing SARS-CoV-2 RNA during a time of low infection prevalence. In an attempt to increase testing capacity, we further adapted our robotic extraction assay to process pooled saliva samples. We also benchmarked our assay against nasopharyngeal swab specimens and found saliva methods require further optimization to match this gold standard. Finally, we designed and validated a RT-qPCR test suitable for saliva self-collection. These results establish a robotic extraction-based procedure for rapid PCR-based saliva testing that is suitable for samples from both symptomatic and asymptomatic individuals.


Assuntos
Teste para COVID-19/métodos , RNA Viral/isolamento & purificação , SARS-CoV-2/genética , Adulto , COVID-19/diagnóstico , Feminino , Humanos , Masculino , Programas de Rastreamento/métodos , RNA/genética , RNA/isolamento & purificação , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real/métodos , Robótica/métodos , Saliva/química , Manejo de Espécimes/métodos
3.
Sci Rep ; 11(1): 16430, 2021 08 12.
Artigo em Inglês | MEDLINE | ID: mdl-34385527

RESUMO

Until there is an effective implementation of COVID-19 vaccination program, a robust testing strategy, along with prevention measures, will continue to be the most viable way to control disease spread. Such a strategy should rely on disparate diagnostic tests to prevent a slowdown in testing due to lack of materials and reagents imposed by supply chain problems, which happened at the beginning of the pandemic. In this study, we have established a single-tube test based on RT-LAMP that enables the visual detection of less than 100 viral genome copies of SARS-CoV-2 within 30 min. We benchmarked the assay against the gold standard test for COVID-19 diagnosis, RT-PCR, using 177 nasopharyngeal RNA samples. For viral loads above 100 copies, the RT-LAMP assay had a sensitivity of 100% and a specificity of 96.1%. Additionally, we set up a RNA extraction-free RT-LAMP test capable of detecting SARS-CoV-2 directly from saliva samples, albeit with lower sensitivity. The saliva was self-collected and the collection tube remained closed until inactivation, thereby ensuring the protection of the testing personnel. As expected, RNA extraction from saliva samples increased the sensitivity of the test. To lower the costs associated with RNA extraction, we performed this step using an alternative protocol that uses plasmid DNA extraction columns. We also produced the enzymes needed for the assay and established an in-house-made RT-LAMP test independent of specific distribution channels. Finally, we developed a new colorimetric method that allowed the detection of LAMP products by the visualization of an evident color shift, regardless of the reaction pH.


Assuntos
Teste para COVID-19/métodos , COVID-19/virologia , Colorimetria/métodos , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , SARS-CoV-2/isolamento & purificação , COVID-19/diagnóstico , Humanos , Pandemias , Portugal/epidemiologia , RNA Viral/genética , SARS-CoV-2/genética , Saliva/química , Saliva/virologia , Sensibilidade e Especificidade
4.
Molecules ; 26(16)2021 Aug 13.
Artigo em Inglês | MEDLINE | ID: mdl-34443499

RESUMO

Over the last years, diverse commercial resin-based composites have dominated as dental filling materials. The purpose of the present study was to determine organic and inorganic eluates from five restorative materials using GC/MS and ICP-OES and to compare the effect on cell survival of human gingival fibroblasts of a conventional and a bioactive resin. Five commercially available restorative materials were employed for this study: ActivaTM Bioactive Restorative, ENA HRi, Enamel plus HRi Biofunction, Fuji II LC Capsule, and Fuji IX Capsule. Disks that were polymerized with a curing LED light or left to set were immersed in: 1 mL methanol or artificial saliva for GC/MS analysis, 5mL deionized water for ICP-OES, and 5mL of culture medium for cell viability. Cell viability was investigated with a modified staining sulforhodamine B assay.The following organic substances were detected: ACP, BHT, BPA, 1,4-BDDMA, CQ, DBP, DMABEE, HEMA, MCE, MeHQ, MOPA, MS, TMPTMA, and TPSb and the ions silicon, aluminum, calcium, sodium, and barium. Activa Bioactive Restorative was found to be biocompatible. Elution of organic substances depended on material's composition, the nature of the solvent and the storage time. Ions' release depended on material's composition and storage time. The newly introduced bioactive restorative was found to be more biocompatible.


Assuntos
Restauração Dentária Permanente , Fibroblastos/citologia , Compostos Inorgânicos/toxicidade , Compostos Orgânicos/toxicidade , Sobrevivência Celular/efeitos dos fármacos , Fibroblastos/efeitos dos fármacos , Cromatografia Gasosa-Espectrometria de Massas , Cimentos de Ionômeros de Vidro/análise , Humanos , Íons , Metanol , Resinas Sintéticas/análise , Saliva/química
5.
J Chromatogr A ; 1652: 462355, 2021 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-34233246

RESUMO

Polyamine metabolites provide pathophysiological information on disease or therapeutic efficacy, yet rapid screening methods for these biomarkers are lacking. Here, we developed high-throughput polyamine metabolite profiling based on multisegment injection capillary electrophoresis triple quadrupole tandem mass spectrometry (MSI-CE-MS/MS), which allows sequential 40-sample injection followed by electrophoretic separation and specific mass detection. To achieve consecutive analysis of polyamine samples, 1 M formic acid was used as the background electrolyte (BGE). The BGE spacer volume had an apparent effect on peak resolution among samples, and 20 nL was selected as the optimal volume. The use of polyamine isotopomers as the internal standard enabled the correction of matrix effects in MS detection. This method is sensitive, selective and quantitative, and its utility was demonstrated by screening polyamines in 359 salivary samples within 360 min, resulting in discrimination of colorectal cancer patients from noncancer controls.


Assuntos
Neoplasias Colorretais/diagnóstico , Eletroforese Capilar/métodos , Poliaminas/análise , Saliva/química , Espectrometria de Massas em Tandem/métodos , Biomarcadores Tumorais/análise , Biomarcadores Tumorais/isolamento & purificação , Neoplasias Colorretais/química , Humanos , Poliaminas/isolamento & purificação
6.
Molecules ; 26(14)2021 Jul 06.
Artigo em Inglês | MEDLINE | ID: mdl-34299389

RESUMO

Currently, most clinical studies in metabolomics only consider a single type of sample such as urine, plasma, or feces and use a single analytical platform, either NMR or MS. Although some studies have already investigated metabolomics data from multiple fluids, the information is limited to a unique analytical platform. On the other hand, clinical studies investigating the human metabolome that combine multi-analytical platforms have focused on a single biofluid. Combining data from multiple sample types for one patient using a multimodal analytical approach (NMR and MS) should extend the metabolome coverage. Pre-analytical and analytical phases are time consuming. These steps need to be improved in order to move into clinical studies that deal with a large number of patient samples. Our study describes a standard operating procedure for biological specimens (urine, blood, saliva, and feces) using multiple platforms (1H-NMR, RP-UHPLC-MS, and HILIC-UHPLC-MS). Each sample type follows a unique sample preparation procedure for analysis on a multi-platform basis. Our method was evaluated for its robustness and was able to generate a representative metabolic map.


Assuntos
Sangue/metabolismo , Fezes/química , Metaboloma , Saliva/química , Manejo de Espécimes/normas , Urina/química , Cromatografia Líquida de Alta Pressão/métodos , Humanos , Espectroscopia de Ressonância Magnética/métodos
7.
Cochrane Database Syst Rev ; 7: CD010276, 2021 07 20.
Artigo em Inglês | MEDLINE | ID: mdl-34282854

RESUMO

BACKGROUND: Squamous cell carcinoma is the most common form of malignancy of the oral cavity, and is often proceeded by oral potentially malignant disorders (OPMD). Early detection of oral cavity squamous cell carcinoma (oral cancer) can improve survival rates. The current diagnostic standard of surgical biopsy with histology is painful for patients and involves a delay in order to process the tissue and render a histological diagnosis; other diagnostic tests are available that are less invasive and some are able to provide immediate results. This is an update of a Cochrane Review first published in 2015. OBJECTIVES: Primary objective: to estimate the diagnostic accuracy of index tests for the detection of oral cancer and OPMD, in people presenting with clinically evident suspicious and innocuous lesions. SECONDARY OBJECTIVE: to estimate the relative accuracy of the different index tests. SEARCH METHODS: Cochrane Oral Health's Information Specialist searched the following databases: MEDLINE Ovid (1946 to 20 October 2020), and Embase Ovid (1980 to 20 October 2020). The US National Institutes of Health Ongoing Trials Register (ClinicalTrials.gov) and the World Health Organization International Clinical Trials Registry Platform were also searched for ongoing trials to 20 October 2020. No restrictions were placed on the language or date of publication when searching the electronic databases. We conducted citation searches, and screened reference lists of included studies for additional references. SELECTION CRITERIA: We selected studies that reported the diagnostic test accuracy of the following index tests when used as an adjunct to conventional oral examination in detecting OPMD or oral cavity squamous cell carcinoma: vital staining (a dye to stain oral mucosa tissues), oral cytology, light-based detection and oral spectroscopy, blood or saliva analysis (which test for the presence of biomarkers in blood or saliva). DATA COLLECTION AND ANALYSIS: Two review authors independently screened titles and abstracts for relevance. Eligibility, data extraction and quality assessment were carried out by at least two authors, independently and in duplicate. Studies were assessed for methodological quality using the Quality Assessment of Diagnostic Accuracy Studies 2 (QUADAS-2). Meta-analysis was used to combine the results of studies for each index test using the bivariate approach to estimate the expected values of sensitivity and specificity. MAIN RESULTS: This update included 63 studies (79 datasets) published between 1980 and 2020 evaluating 7942 lesions for the quantitative meta-analysis. These studies evaluated the diagnostic accuracy of conventional oral examination with: vital staining (22 datasets), oral cytology (24 datasets), light-based detection or oral spectroscopy (24 datasets). Nine datasets assessed two combined index tests. There were no eligible diagnostic accuracy studies evaluating blood or salivary sample analysis. Two studies were classed as being at low risk of bias across all domains, and 33 studies were at low concern for applicability across the three domains, where patient selection, the index test, and the reference standard used were generalisable across the population attending secondary care. The summary estimates obtained from the meta-analysis were: - vital staining: sensitivity 0.86 (95% confidence interval (CI) 0.79 to 0.90) specificity 0.68 (95% CI 0.58 to 0.77), 20 studies, sensitivity low-certainty evidence, specificity very low-certainty evidence; - oral cytology: sensitivity 0.90 (95% CI 0.82 to 0.94) specificity 0.94 (95% CI 0.88 to 0.97), 20 studies, sensitivity moderate-certainty evidence, specificity moderate-certainty evidence; - light-based: sensitivity 0.87 (95% CI 0.78 to 0.93) specificity 0.50 (95% CI 0.32 to 0.68), 23 studies, sensitivity low-certainty evidence, specificity very low-certainty evidence; and - combined tests: sensitivity 0.78 (95% CI 0.45 to 0.94) specificity 0.71 (95% CI 0.53 to 0.84), 9 studies, sensitivity very low-certainty evidence, specificity very low-certainty evidence. AUTHORS' CONCLUSIONS: At present none of the adjunctive tests can be recommended as a replacement for the currently used standard of a surgical biopsy and histological assessment. Given the relatively high values of the summary estimates of sensitivity and specificity for oral cytology, this would appear to offer the most potential. Combined adjunctive tests involving cytology warrant further investigation. Potentially eligible studies of blood and salivary biomarkers were excluded from the review as they were of a case-control design and therefore ineligible. In the absence of substantial improvement in the tests evaluated in this updated review, further research into biomarkers may be warranted.


Assuntos
Carcinoma de Células Escamosas/diagnóstico , Neoplasias Bucais/diagnóstico , Viés , Biomarcadores Tumorais/análise , Biomarcadores Tumorais/sangue , Carcinoma de Células Escamosas/patologia , Corantes , Detecção Precoce de Câncer , Humanos , Luz , Neoplasias Labiais/diagnóstico , Neoplasias Labiais/patologia , Boca/patologia , Neoplasias Bucais/patologia , Saliva/química , Sensibilidade e Especificidade
8.
J Forensic Sci ; 66(5): 1871-1878, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34287912

RESUMO

Oral fluid is a valuable alternative matrix for forensic toxicologists due to ease of observed collection, limited biohazardous exposure, and indications of recent drug use. Limited information is available for fentanyl analog prevalence, interpretation, or analysis in oral fluid. With increasing numbers of fentanyl-related driving under the influence of drug (DUID) cases appearing in the United States, the development of detection methods is critical. The purpose of the present study was to develop and validate a quantitative method for fentanyl analogs in oral fluid (collected via Quantisal™) using liquid chromatography-quadrupole-time-of-flight-mass spectrometry (LC-QTOF-MS). Validation resulted in limits of detection and quantification ranging from 0.5 to 1 ng/mL. Established linear range was 1-100 ng/mL for all analytes, except acetyl fentanyl at 0.5-100 ng/mL (R2  > 0.994). Within- and between-run precision and bias were considered acceptable with maximum values of ±15.2%CV and ±14.1%, respectively. Matrix effects exhibited ionization enhancement for all analytes with intensified enhancement at a low concentration (9.3-47.4%). No interferences or carryover was observed. Fentanyl analogs were stable in processed extracts stored in the autosampler (4° C) for 48h. The validated method was used to quantify fentanyl analogs in authentic oral fluid samples (n=17) from probationers/parolees. Fentanyl and 4-ANPP concentrations were 1.0-104.5 ng/mL and 1.2-5.7 ng/mL, respectively.


Assuntos
Fentanila/análogos & derivados , Fentanila/análise , Saliva/química , Analgésicos Opioides/análise , Cromatografia Líquida , Toxicologia Forense/métodos , Humanos , Espectrometria de Massas/métodos , Detecção do Abuso de Substâncias/métodos
9.
Turk J Pediatr ; 63(3): 404-416, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34254485

RESUMO

BACKGROUND: Recurrent wheezing is common in young children, with a cumulative prevalence of up to 40 % in the first 6 years of life. In this study, we aimed to evaluate the relationship between the number of wheezing episodes and the number of cigarettes smoked at home and serum / saliva cotinine and carnosine levels in children with recurrent wheezing. METHODS: This study was conducted with 80 young children with recurrent wheezing, aged between 1-4 years and 50 healthy control groups. Patient population was divided into three groups depending on the number of their exposure to cigarette smoke and wheezing attacks. Serum cotinine, saliva cotinine, serum carnosine, saliva carnosine, vitamin D levels were measured by using the ELISA method. RESULTS: A significant relationship for serum cotinine and saliva cotinine levels was found between groups (p < 0.05). It was determined that as the number of exposure to cigarette smoke and number of wheezing episodes in young children with recurrent wheezing increased, the level of serum/saliva cotinine levels increased significantly, compared to the control group. In contrast, it was determined that as the number of exposure to cigarette smoke and number of wheezing episodes in young children with recurrent wheezing increased, serum/saliva carnosine levels decreased significantly, compared to the control group. In addition, a significant difference in serum vitamin D levels was found between healthy young children and young children with recurrent wheezing (p < 0.05). CONCLUSIONS: We think that the measurement of salivary cotinine is a useful and noninvasive marker to evaluate passive smoking exposure in the etiology of recurrent wheezing in young children.


Assuntos
Carnosina , Poluição por Fumaça de Tabaco , Criança , Pré-Escolar , Cotinina/análise , Humanos , Lactente , Sons Respiratórios/etiologia , Saliva/química , Poluição por Fumaça de Tabaco/efeitos adversos
10.
Int J Mol Sci ; 22(13)2021 Jun 29.
Artigo em Inglês | MEDLINE | ID: mdl-34209954

RESUMO

Several studies have highlighted the diagnostic potential of salivary microRNA (miRNA) in head and neck squamous cell cancer (HNSCC). The purpose of this meta-analysis was to summarize published studies and evaluate the diagnostic accuracy of salivary miRNA in HNSCC detection. In this meta-analysis, we systematically searched PubMed, EMBASE, and Cochrane Library databases for studies on miRNA and HNSCC diagnosis. Pooled sensitivity, specificity, and diagnostic odds ratio (DOR) with a summary receiver-operating characteristic curve were calculated using a bivariate random-effect meta-analysis model. Furthermore, subgroup analyses were conducted to explore the main sources of heterogeneity. Seventeen studies from ten articles, including 23 miRNA and a total of 759 subjects, were included in this meta-analysis. The pooled sensitivity and specificity of salivary miRNA in the diagnosis of HNSCC were 0.697 (95% CI: 0.644-0.744) and 0.868 (95% CI: 0.811-0.910), respectively. The overall area under the curve was 0.803 with a DOR of 12.915 (95% CI: 9.512-17.534). Salivary miRNAs are a promising non-invasive diagnostic biomarker with moderate accuracy for HNSCC. These results must be verified by large-scale prospective studies.


Assuntos
Neoplasias de Cabeça e Pescoço/diagnóstico , MicroRNAs/genética , Saliva/química , Carcinoma de Células Escamosas de Cabeça e Pescoço/diagnóstico , Biomarcadores Tumorais/genética , Detecção Precoce de Câncer , Neoplasias de Cabeça e Pescoço/genética , Humanos , Razão de Chances , Sensibilidade e Especificidade , Carcinoma de Células Escamosas de Cabeça e Pescoço/genética
11.
Vet J ; 273: 105679, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34148602

RESUMO

Salivary biomarkers were studied in 17 healthy Large White sows from early gestation to the end of lactation. Saliva samples were obtained at 34 ± 3 days from insemination (G30), 24 ± 4 days before farrowing (G90), within the first 24 h after farrowing (L1) and at the end of a lactation period of 21 days (L21). The measurements in saliva included stress-related biomarkers (cortisol, chromogranin A, α-amylase, butyrylcholinesterase [BChE] and lipase [Lip]), inflammatory biomarkers (adenosine deaminase isoenzymes 1 [ADA1] and 2 [ADA2], and haptoglobin [Hp]) and oxidative stress biomarkers (cupric reducing antioxidant capacity, trolox equivalent antioxidant capacity, ferric reducing ability, uric acid, advanced oxidation protein products [AOPP] and hydrogen peroxide [H2O2]), as well as routine biochemistry analytes (aspartate aminotransferase [AST], alkaline phosphatase [ALP], γ-glutamine transferase [GGT], lactate dehydrogenase [LDH], creatine kinase [CK], urea, creatinine, triglycerides, lactate, calcium and phosphorus). The main changes were observed at farrowing, with increases in biomarkers of stress (cortisol and BChE), inflammation (ADA isoenzymes and Hp) and oxidative stress (AOPP and H2O2), as well as muscle and hepatic enzymes (CK, AST, ALP, GGT and LDH). Lactate and triglycerides increased at the end of gestation and remained at high concentrations until the end of lactation. Lip was higher in gestation than at lactation. Thus, changes in biomarkers of stress, immune function, oxidative stress, hepatic and muscle integrity, and energy mobilization occur in sow saliva during pregnancy, farrowing and lactation. These changes, caused by physiological conditions, should be taken into consideration when these biomarkers are used for the evaluation of sow health and welfare.


Assuntos
Biomarcadores/análise , Lactação/fisiologia , Gravidez/fisiologia , Saliva/química , Sus scrofa/fisiologia , Animais , Metabolismo Energético , Feminino , Inflamação , Estresse Oxidativo , Parto/fisiologia , Saliva/enzimologia
12.
Health Psychol ; 40(5): 316-325, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-34152785

RESUMO

OBJECTIVE: Lesbian, gay, and bisexual (LGB) individuals who report greater minority stress (e.g., discrimination) are at an elevated risk for multiple health problems. However, few studies have examined physiological mechanisms that might link minority stress to health. This study tested how cardiovascular and cortisol responses to a laboratory-induced social stressor differed when that stressor contained an additional minority stress component. METHOD: LGB adults (n = 141; 51% male, 49% female) participated in a social stress task in which they were interviewed by a prerecorded confederate. Participants were randomized to receive information that their interviewer held either antigay or progay social/political beliefs. Cardiovascular reactivity and salivary cortisol were assessed at baseline, during the task, and during recovery. RESULTS: All participants experienced significant task-related increases in heart rate (HR), systolic blood pressure (SBP), and diastolic blood pressure (DBP). However, participants in the antigay condition had greater increases in HR and SBP during the task and smaller decreases in SBP during recovery. Salivary cortisol increased significantly only in the antigay condition. High frequency heart rate variability (hfHRV) was constant throughout the stress task for participants in the progay condition but decreased significantly during the task for participants in the antigay condition. CONCLUSIONS: Minority stress has the potential to affect LGB individuals' health through cardiovascular and endocrine mechanisms. Moreover, its physiological signature may differ from other social stress in ways that have implications for health and emotion regulation more broadly. (PsycInfo Database Record (c) 2021 APA, all rights reserved).


Assuntos
Pressão Sanguínea/fisiologia , Frequência Cardíaca/fisiologia , Hidrocortisona/metabolismo , Minorias Sexuais e de Gênero/psicologia , Estresse Psicológico/fisiopatologia , Adolescente , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Saliva/química , Minorias Sexuais e de Gênero/estatística & dados numéricos , Adulto Jovem
13.
Artigo em Inglês | MEDLINE | ID: mdl-34069235

RESUMO

Smoke-free home rules restrict smoking in the home, but biomarkers of secondhand smoke exposure are needed to help understand the association between smoke-free homes and child secondhand smoke exposure. Participants (n = 346) were majority Black/African American mother-child dyads from a longitudinal study in North Carolina. Mothers completed questionnaires on household smoking behaviors and rules, and child saliva samples were assayed for secondhand smoke exposure. Regression models used smoke-free home rules to predict child risk for secondhand smoke exposure. Children in households with smoke-free home rules had less salivary cotinine and risk for secondhand smoke exposure. After controlling for smokers in the household, home smoking rules were not a significant predictor of secondhand smoke exposure. Compared to children in households with no smokers, children in households with at least one smoker but a non-smoking mother (OR 5.35, 95% CI: 2.22, 13.17) and households with at least one smoker including a smoking mother (OR 13.73, 95% CI: 6.06, 33.28) had greater risk for secondhand smoke exposure. Results suggest smoke-free home rules are not sufficient to fully protect children from secondhand smoke exposure, especially in homes with smokers. Future research should focus on how household members who smoke can facilitate the prevention of child secondhand smoke exposure.


Assuntos
Poluição por Fumaça de Tabaco , Criança , Feminino , Humanos , Estudos Longitudinais , Relações Mãe-Filho , North Carolina , Saliva/química , Poluição por Fumaça de Tabaco/análise
14.
Medicine (Baltimore) ; 100(25): e26369, 2021 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-34160409

RESUMO

BACKGROUND: Diseases of the oral cavity (OC) with an infectious trigger such as caries and periodontal disease are extremely common in the general population and can also have effects at the cardiovascular level. The oral salivary flow, with its buffering capacity, is able to regulate the pH of the OC and, therefore, significantly contribute to the ecological balance of the microenvironment in which the oral microbiome (OM) develops. On the other side, when the quality/quantity of salivary flow is altered it is supposed the disruption of this balance with the potential increase in oral pathogens and triggered diseases. Among the endogenous substances able to exert a significant effect on the salivary flow and its characteristics, carnosine (Car), a dipeptide originally isolated in skeletal muscle, represents, thanks to the known buffering properties, a promising principle. METHODS: We aimed this protocol to evaluate the quantitative/qualitative characteristics of the salivary flow in healthy volunteer subjects (n = 20) and in subjects suffering from common OC pathologies (n = 40), before and after 7 days of supplementation with SaliflussTM (Metis Healthcare srl, Milan, Italy), a Class I medical device on the market as 400 mg mucoadhesive oral tablets that has Car as the main ingredient. DISCUSSION: Combining the characteristics of saliva with the OM and comparing them with OC pathologies, we expect to clarify their reciprocal relationship and, using quantitative proteomics techniques, to help clarify the mechanism of action of Car.


Assuntos
Carnosina/administração & dosagem , Cárie Dentária/dietoterapia , Gengivite/dietoterapia , Periodontite/dietoterapia , Saliva/química , Administração Bucal , Adolescente , Adulto , Doenças Cardiovasculares/etiologia , Doenças Cardiovasculares/prevenção & controle , Cárie Dentária/complicações , Cárie Dentária/prevenção & controle , Suplementos Nutricionais , Gengivite/microbiologia , Gengivite/prevenção & controle , Voluntários Saudáveis , Humanos , Concentração de Íons de Hidrogênio , Masculino , Microbiota/fisiologia , Mucosa Bucal/microbiologia , Periodontite/microbiologia , Periodontite/prevenção & controle , Saliva/metabolismo , Comprimidos , Adulto Jovem
15.
J Biol Chem ; 297(1): 100865, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-34118237

RESUMO

During feeding, a tick's mouthpart penetrates the host's skin and damages tissues and small blood vessels, triggering the extrinsic coagulation and lectin complement pathways. To elude these defense mechanisms, ticks secrete multiple anticoagulant proteins and complement system inhibitors in their saliva. Here, we characterized the inhibitory activities of the homologous tick salivary proteins tick salivary lectin pathway inhibitor, Salp14, and Salp9Pac from Ixodesscapularis in the coagulation cascade and the lectin complement pathway. All three proteins inhibited binding of mannan-binding lectin to the polysaccharide mannan, preventing the activation of the lectin complement pathway. In contrast, only Salp14 showed an appreciable effect on coagulation by prolonging the lag time of thrombin generation. We found that the anticoagulant properties of Salp14 are governed by its basic tail region, which resembles the C terminus of tissue factor pathway inhibitor alpha and blocks the assembly and/or activity of the prothrombinase complex in the same way. Moreover, the Salp14 protein tail contributes to the inhibition of the lectin complement pathway via interaction with mannan binding lectin-associated serine proteases. Furthermore, we identified BaSO4-adsorbing protein 1 isolated from the tick Ornithodoros savignyi as a distant homolog of tick salivary lectin pathway inhibitor/Salp14 proteins and showed that it inhibits the lectin complement pathway but not coagulation. The structure of BaSO4-adsorbing protein 1, solved here using NMR spectroscopy, indicated that this protein adopts a noncanonical epidermal growth factor domain-like structural fold, the first such report for tick salivary proteins. These data support a mechanism by which tick saliva proteins simultaneously inhibit both the host coagulation cascade and the lectin complement pathway.


Assuntos
Proteínas de Artrópodes/ultraestrutura , Interações Hospedeiro-Patógeno/genética , Lectinas/genética , Proteínas e Peptídeos Salivares/ultraestrutura , Animais , Proteínas de Artrópodes/química , Proteínas de Artrópodes/genética , Coagulação Sanguínea/genética , Vasos Sanguíneos/parasitologia , Vasos Sanguíneos/patologia , Lectina de Ligação a Manose da Via do Complemento/genética , Ixodes/patogenicidade , Ixodes/ultraestrutura , Lectinas/ultraestrutura , Espectroscopia de Ressonância Magnética , Conformação Proteica , Saliva/química , Saliva/metabolismo , Proteínas e Peptídeos Salivares/química , Proteínas e Peptídeos Salivares/genética , Trombina/genética , Carrapatos/genética , Carrapatos/patogenicidade
16.
J Alzheimers Dis ; 82(3): 899-904, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34120906

RESUMO

Diurnal salivary cortisol was measured in 334 older adults without dementia, at four times on two separate days, under quiet and stressful conditions. In multivariate Cox proportional hazard models, higher global diurnal cortisol secretion was associated with incident dementia (HR = 1.09 [1.02-1.15] per one-unit increase in cortisol measure, p = 0.007) and Alzheimer's disease (HR = 1.12 [1.04-1.21], p = 0.003) over a mean (SD) of 8.1 (4.0) years, independent of potential confounders and stressful conditions. Individuals with incident dementia had a slower rate of cortisol elimination under non-stressful conditions, reflected by higher cortisol levels in the evening, and an abnormal response to stress (blunted evening stress response).


Assuntos
Ritmo Circadiano/fisiologia , Demência/metabolismo , Demência/psicologia , Hidrocortisona/metabolismo , Achados Incidentais , Vida Independente/psicologia , Idoso , Idoso de 80 Anos ou mais , Demência/diagnóstico , Feminino , Seguimentos , Humanos , Hidrocortisona/análise , Vida Independente/tendências , Masculino , Sintomas Prodrômicos , Estudos Prospectivos , Saliva/química , Saliva/metabolismo , Estresse Psicológico/diagnóstico , Estresse Psicológico/metabolismo , Estresse Psicológico/psicologia
17.
FASEB J ; 35(8): e21745, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-34191346

RESUMO

Studies are needed to identify useful biomarkers to assess the severity and prognosis of COVID-19 disease, caused by severe acute respiratory syndrome coronavirus (SARS-CoV-2) virus. Here, we examine the levels of various plasma species of the SARS-CoV-2 host receptor, the angiotensin-converting enzyme 2 (ACE2), in patients at different phases of the infection. Human plasma ACE2 species were characterized by immunoprecipitation and western blotting employing antibodies against the ectodomain and the C-terminal domain, using a recombinant human ACE2 protein as control. In addition, changes in the cleaved and full-length ACE2 species were also examined in serum samples derived from humanized K18-hACE2 mice challenged with a lethal dose of SARS-CoV-2. ACE2 immunoreactivity was present in human plasma as several molecular mass species that probably comprise truncated (70 and 75 kDa) and full-length forms (95, 100, 130, and 170 kDa). COVID-19 patients in the acute phase of infection (n = 46) had significantly decreased levels of ACE2 full-length species, while a truncated 70-kDa form was marginally higher compared with non-disease controls (n = 26). Levels of ACE2 full-length species were in the normal range in patients after a recovery period with an interval of 58-70 days (n = 29), while the 70-kDa species decreased. Levels of the truncated ACE2 species served to discriminate between individuals infected by SARS-CoV-2 and those infected with influenza A virus (n = 17). In conclusion, specific plasma ACE2 species are altered in patients with COVID-19 and these changes normalize during the recovery phase. Alterations in ACE2 species following SARS-CoV-2 infection warrant further investigation regarding their potential usefulness as biomarkers for the disease process and to asses efficacy during vaccination.


Assuntos
Enzima de Conversão de Angiotensina 2/sangue , COVID-19/sangue , SARS-CoV-2 , Adulto , Idoso , Idoso de 80 Anos ou mais , Enzima de Conversão de Angiotensina 2/líquido cefalorraquidiano , Enzima de Conversão de Angiotensina 2/química , Enzima de Conversão de Angiotensina 2/urina , Biomarcadores/sangue , Química Encefálica , Colo/química , Feminino , Humanos , Fígado/química , Masculino , Pessoa de Meia-Idade , Saliva/química
18.
J Chromatogr A ; 1651: 462273, 2021 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-34087718

RESUMO

This study presents an accurate and precise analytical strategy for the determination of chloroquine phosphate at trace levels in human body fluids (urine, serum, and saliva). Simultaneous derivatization-spraying based fine droplet formation-liquid phase microextraction (SD-SFDF-LPME) method was used to derivatize and preconcentrate the analyte prior to gas chromatography-mass spectrometry (GC-MS) measurements. Acetic anhydride was employed as derivatizing agent in this study. After optimizing the SD-SFDF-LPME method, the limit of detection (LOD) and limit of quantitation (LOQ) were found to be 0.16 and 0.53 mg/kg, respectively. Quadruple isotope dilution (ID4) was coupled to the SD-SFDF-LPME method in order to alleviate matrix effects and promote accuracy/precision of the method. Chloroquine acetamide-d3 was firstly synthesized in our research laboratory and used as the isotopic analogue of the analyte in the ID4 experiments. Superior percent recovery results (99.4% - 101.0%) with low standard deviation values were obtained for the spiked samples. This validated the developed SD-SFDF-LPME-ID4-GC-MS method as highly accurate and precise for the determination of chloroquine phosphate at trace levels. In addition, the isotopic analogue of the analyte was obtained via the acetamide derivative of the analyte, which is an alternative to obtain isotopic analogues of organic compounds that are not accessible or commercially available.


Assuntos
Cloroquina/análogos & derivados , Cromatografia Gasosa-Espectrometria de Massas/métodos , Microextração em Fase Líquida/métodos , Líquidos Corporais/química , Cloroquina/análise , Cloroquina/sangue , Cloroquina/isolamento & purificação , Cloroquina/urina , Humanos , Isótopos , Limite de Detecção , Saliva/química
19.
J Chromatogr A ; 1651: 462278, 2021 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-34102399

RESUMO

A simple method for the determination of polyamines and their N-acetylated forms was developed using benzoyl chloride as derivatization reagent, and 1,6-diaminohexane as internal standard, followed by liquid-liquid extraction with ethyl acetate. The organic extract was injected in a gas chromatograph using a programmed temperature vaporizer and the determination and quantification was performed with a quadrupole mass spectrometer. There was no matrix effect with the proposed method, so internal calibration was used to quantify the corresponding derivatives. Good linear responses were obtained in the range from the limits of detection to 500 µg L-1 (50 µg L-1 for spermidine), with correlation coefficients varying from 0.9591 to 0.9968. The limits of quantification (S/N = 10) ranged 1.0 - 8.3 µg L-1. Recoveries were found between 82 - 117%, showing the good accuracy of the proposed method. Intra- and inter-day precision assays, expressed as relative standard deviation (RSD) were evaluated at two different concentration levels (low and high), showing values in the range of 2.4 - 6.1% and 5.2 - 9.0% for repeatability and reproducibility, respectively (6.9 - 9.7% and 14.1 - 14.6% for spermidine). Successful determination of the studied polyamines and their N-acetylated forms was performed on the saliva of 17 volunteers.


Assuntos
Cromatografia Gasosa-Espectrometria de Massas/métodos , Poliaminas/análise , Saliva/química , Acetilação , Benzoatos , Diaminas , Humanos , Extração Líquido-Líquido , Poliaminas/química , Reprodutibilidade dos Testes
20.
Nutrients ; 13(5)2021 May 11.
Artigo em Inglês | MEDLINE | ID: mdl-34064929

RESUMO

BACKGROUND: Chronic stress is often accompanied by alterations in the diurnal rhythm of hypothalamus-pituitary-adrenal activity. However, there are limited data on the diurnal rhythmicity of breast milk glucocorticoids (GCs) among women with psychological distress. We compared mothers who sought consultation at an expertise center for pregnant women with an increased risk of psychological distress with control mothers for GC diurnal rhythmicity in milk and saliva obtained at the same time. METHODS: We included 19 mothers who sought consultation at the psychiatry-obstetric-pediatric (POP) outpatient clinic and 44 control mothers. One month postpartum, mothers collected on average eight paired milk and saliva samples during a 24 h period. GC levels were measured using liquid chromatography-tandem mass spectrometry. GC rhythmicity parameters were determined with specialized software. RESULTS: For both milk and saliva, no group differences regarding GC rhythms were found. Milk cortisol area under the curve with respect to the ground was lower in the POP group than in the control group (p = 0.02). GC levels in human milk and saliva were highly correlated within each group (p < 0.001). CONCLUSION: Although there were no differences between groups in GC rhythmicity, the total amount of milk cortisol was lower in the POP group. Long-term follow-up is needed to address the impact of vertical transmission of breast milk GCs.


Assuntos
Ritmo Circadiano , Glucocorticoides/análise , Leite Humano/química , Estresse Psicológico , Adulto , Feminino , Humanos , Hidrocortisona/análise , Mães/psicologia , Gravidez , Gestantes , Psicopatologia , Saliva/química
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