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1.
Pediatrics ; 149(1)2022 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-34907443

RESUMO

OBJECTIVES: Daytime urinary incontinence is disabling and occurs in 17% of school-aged children. Timed-voiding is part of standard therapy. Can an alarm watch to aid timed-voiding improve treatment response to standard therapy? METHODS: The WATCH (Watch with Alarm for Timed-Voiding in Children) study is a randomized controlled trial. Participants were randomly assigned (1:1) to a vibrating alarm or nonalarming watch for 3-months. The primary outcome was the proportion who achieved a complete response (14 consecutive dry days) after 3-months of treatment. Children aged 5 to 13 years who were prescribed timed-voiding for daytime urinary incontinence. RESULTS: Overall, 243 children, with a mean age of 8 years, were enrolled, with 62% girls. At 3-months, the complete response rates were similar between the 2 groups (22% alarm versus 17% control; difference: 5%; 95% confidence interval (CI): -5% to 16%; P = .42). In the alarm group, treatment adherence was higher (40% vs 10%; difference: 30%; 95% CI: 20% to 40%; P < .001), frequency of incontinence was lower (25% dry; 40% had 1-3 wet days per week, 24% had 4-6 wet days per week, and 12% had daily wetting, compared with 19%, 30%, 35%, and 16%, respectively; P =.05), and fewer had abnormal postvoid residual urine volumes (12% vs 24%; difference: -12%; 95% CI: -21% to -1%; P = .04) compared with the control group. Improvement was transient and did not persist 6 months beyond the treatment period. CONCLUSIONS: Alarm watches do not appear to lead to complete resolution of urinary incontinence in children but did promote treatment adherence, normalization of postvoid residual volumes, and reduction in incontinent episodes while being used.


Assuntos
Cooperação do Paciente , Sistemas de Alerta , Incontinência Urinária/prevenção & controle , Adolescente , Criança , Pré-Escolar , Feminino , Seguimentos , Humanos , Masculino , Fatores de Tempo , Resultado do Tratamento , Incontinência Urinária/fisiopatologia , Urina
3.
Clin Lab ; 67(11)2021 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-34758221

RESUMO

BACKGROUND: Automated microscopic platforms are increasingly used in clinical laboratories for rapid analysis of samples. However, it is important to present the results quantitatively or semiquantitatively because automated platforms use various technologies for analysis as well as different sediment preparation methods. The results of cell counting using an on screen image review program for the cobas u 701 analyzer (Roche Diagnostics Interna-tional, Rotkreuz, Switzerland) differed from those obtained by manual microscopic examination (MME). This study was performed to investigate the difference of results among analyzer, on-screen image review and MME. METHODS: Freshly collected urine specimens from outpatients were used. We calculated the mean, standard deviation, and 95% confidence interval for red and white blood cell (RBC/WBC) quantitative results obtained using the cobas u 701 analyzer. These results were compared to those obtained by manual counting. RBC and WBC counts determined with the cobas u 701 analyzer were compared to those obtained by MME per unit field. RESULTS: The semiquantitative results of MME were graded as 0 - 2, 3 - 5, 6 - 10, 11 - 20, 21 - 30, and many or numerous cells/high power field (HPF). The RBC and WBC counts determined by image analyses showed the tendency to be one grade higher than those from MME in the range of 3 to 5/HPF to many/HPF. The results of nearly all samples with 0 - 2/HPF and numerous/HPF for RBC and WBC counts were consistent with the grade found by MME. CONCLUSIONS: The one-grade difference may have been caused by the differences of preanalytical factors in the sample volume, centrifugal force, urine concentration ratio, or sediment volume/area of the slide. When reporting the results of image analyses, RBC and WBC counts should be raised by one grade to compensate for MME. Each laboratory needs to verify the on-screen review of images corresponding to the microscopic field of view according to the clinical laboratory's specific preanalytical practices.


Assuntos
Microscopia , Urinálise , Laboratórios , Contagem de Leucócitos , Leucócitos , Urina
4.
Pan Afr Med J ; 40: 75, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34804342

RESUMO

Introduction: metabolic abnormalities are key factors in urolithiasis patients because they can be modified to prevent the risk of urinary stones. The objectives of this study were to estimate the frequency of metabolic abnormalities in the urine of patients with urolithiasis and to determine their possible link with the chemical composition of stones. Methods: we conducted a cross-sectional study evaluating 73 patients referred for urolithiasis in 8 clinics in Kinshasa, between January 2017 and September 2019. Twenty four-hour or early morning urine were collected and analyzed in the Tenon Hospital in Paris. Parameters analyzed included pH, specific gravity, creatinine, uric acid, calcium, phosphate, oxalate, citrate and magnesium. Chi square test or chi-square likelihood-ratio and student's t test were used as statistical tests. Results: overall, 89% (n=65) of patients with lithiasis had metabolic abnormalities. Mean (SD) age of patients was 47.0 (14.2) years with male to female ratio of 1.6: 1. The mean (SD) 24-hour diuresis was 1836.4 (1216.9) ml; the mean (SD) urine density was 1.018 (0.007); and the mean (SD) pH was 6.1(0.8). Hypocitraturia was the most frequently observed metabolic abnormality and was found in 76.7% patients. Other significant metabolic abnormalities were low magnesuria (35.6%), hyperoxaluria (11%), and low sulphaturia (74%). Whewellite (73.5%) was the main chemical component. The mean pH was higher in patients with carbapatite and struvite stones (p=0.031). Conclusion: this study suggests that inadequate diuresis and hypocitraturia were important lithogenic factors. The population should be encouraged to increase water intake to limit the frequency of urine super saturation with crystals.


Assuntos
Cálculos Urinários/química , Urina/química , Urolitíase/epidemiologia , Adulto , Ácido Cítrico/urina , Estudos Transversais , República Democrática do Congo , Diurese/fisiologia , Feminino , Humanos , Concentração de Íons de Hidrogênio , Masculino , Pessoa de Meia-Idade , Prevalência , Fatores de Risco , Adulto Jovem
5.
Washington, D.C.; OPS; 2021-11-15.
em Espanhol | PAHO-IRIS | ID: phr-55189

RESUMO

Esta guía práctica tiene como objetivo ayudar a los países a implementar e incorporar la prueba Alere Determine™ TB LAM Ag (antígeno) en su algoritmo diagnóstico habitual para la tuberculosis (TB). Contiene nueva evidencia recopilada de numerosos estudios realizados después de que se publicara la primera política de la Organización Mundial de la Salud (OMS) sobre el uso de la prueba de determinación del lipoarabinomanano en orina mediante inmunocromatografía de flujo lateral (LAM-ICL) en el 2015 y se actualizara en noviembre del 2019. La prueba LAM-ICL ayudará a diagnosticar la TB activa en adultos, adolescentes y niños con infección por el VIH. En esta guía se incorporan nuevas recomendaciones sobre la inclusión de la prueba en los algoritmos de diagnóstico, teniendo en cuenta criterios específicos para las pruebas en entornos hospitalarios y ambulatorios, y también contiene información más detallada sobre el proceso de laboratorio para realizar la prueba y sobre las enseñanzas extraídas, lo que ayudará a orientar a los países para que lleven a cabo una implementación más rápida y sencilla de la prueba. El público destinatario incluye a los responsables de la formulación de políticas nacionales y subnacionales, al personal de salud de primera línea y los equipos directivos de los programas de VIH y tuberculosis, y a las principales partes interesadas en la tuberculosis, incluidos los asociados en la ejecución.


Assuntos
HIV , Cromatografia de Afinidade , Tuberculose , Doenças Transmissíveis , Urina , Infecções por HIV , Linfocitose , Contagem de Linfócito CD4 , Gravidade do Paciente , Teste de HIV , Mycobacterium tuberculosis
6.
J Ethnobiol Ethnomed ; 17(1): 62, 2021 Oct 28.
Artigo em Inglês | MEDLINE | ID: mdl-34711254

RESUMO

BACKGROUND: Despite a widespread aversion towards faeces and urine, animal excreta are used in traditional medicine in many countries since centuries, but records are scattered and few therapeutic uses have been accurately documented while in the current context of emerging zoonoses such records may be of major interest. METHODOLOGY: In this study, we investigated the therapeutic uses that mahouts in Xayaboury province, Lao PDR make of elephant urine and faeces as well as of the brood chamber that beetles (Heliocopris dominus) fashion from elephant dung. Semi-structured interviews were conducted with mahouts on elephant diet, health problems and responses to disease, andwhether they use elephant products. Data were supplemented by interviews with traditional healers. RESULTS: Seven respondents reported the use of elephant urine in ethnoveterinary care for elephants and in human medicine in case of diabetes and otitis. 25 respondents reported therapeutic use of elephant faeces (EF) and elephant dung beetle brood chambers. The major indications are gastrointestinal and skin problems. Macerations or decoctions are drunk or used externally as a lotion. The mahouts attribute the therapeutic effectiveness of EFs to their content which includes the remains of many species from the elephant diet which they consider to be medicinal. DISCUSSION: The indications of these uses are consistent with pharmacological and clinical studies highlighting the properties of different animals' urine and faeces and their curative potential tested in vivo. The acknowledgement by the mahouts of medicinal properties of elephant faecal bolus contrasts with the rare justifications of animal material use recorded in zootherapeutic studies, which falls within the symbolic domain. However, numerous studies highlight the preponderant role of the microbiota in physiological processes, raising the hypothesis of a curative action of EF, by rebalancing the user's microbiota. CONCLUSION: The therapeutic uses of EF preparations despite their possible curative properties are a potential source of zoonotic transmission from elephants to humans. In the current context of globalisation of trade which favours the emergence of zoonoses and in relation with the issue of One Health, it becomes crucial to further document the zootherapeutic practices to prevent emerging diseases. As elephants and local related ethnoethological knowledge are threatened, documenting them is urgent to contribute to their preservation.


Assuntos
Elefantes , Fezes , Medicina Tradicional , Urina , Animais , Laos
7.
Environ Sci Technol ; 55(21): 14886-14896, 2021 11 02.
Artigo em Inglês | MEDLINE | ID: mdl-34637289

RESUMO

Recovering nitrogen from source-separated urine is an important part of the sustainable nitrogen management. A novel bipolar membrane electrodialysis with membrane contactor (BMED-MC) process is demonstrated here for efficient recovery of ammonia from synthetic source-separated urine (∼3772 mg N L-1). In a BMED-MC process, electrically driven water dissociation in a bipolar membrane simultaneously increases the pH of the urine stream and produces an acid stream for ammonia stripping. With the increased pH of urine, ammonia transports across the gas-permeable membrane in the membrane contactor and is recovered by the acid stream as ammonium sulfate that can be directly used as fertilizer. Our results obtained using batch experiments demonstrate that the BMED-MC process can achieve 90% recovery. The average ammonia flux and the specific energy consumption can be regulated by varying the current density. At a current density of 20 mA cm-2, the energy required to achieve a 67.5% ammonia recovery in a 7 h batch mode is 92.8 MJ kg-1 N for a bench-scale system with one membrane stack and can approach 25.8 MJ kg-1 N for large-scale systems with multiple membrane stacks, with an average ammonia flux of 2.2 mol m-2 h-1. Modeling results show that a continuous BMED-MC process can achieve a 90% ammonia recovery with a lower energy consumption (i.e., 12.5 MJ kg-1 N). BMED-MC shows significant potential for ammonia recovery from source-separated urine as it is relatively energy-efficient and requires no external acid solution.


Assuntos
Amônia , Nitrogênio , Fertilizantes , Urina
8.
Clin Lab ; 67(10)2021 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-34655212

RESUMO

BACKGROUND: Urinalysis is used as a screening tool in case of a clinical suspicion of urinary tract infection (UTI). The purpose of this study is to evaluate the analytical performance, agreement, and screening value of four commercial point-of-care testing (POCT) urine analyzers for UTI. METHODS: The following four POCT analyzers were evaluated: Clinitek Status+, BW-200, Hipee S2, and URS-14K. The agreement of the four analyzers with the laboratory standard was assessed. The diagnostic indicators of the screening tests performed using POCT analyzers for UTI were calculated against the bacterial culture as the gold standard. RESULTS: Overall agreement of the four analyzers was good. For nitrite and leukocytes, all the analyzers showed substantial or excellent agreement with the laboratory reference standard, whereas agreement for erythrocytes varied from moderate to excellent. With each POCT analyzer, combined detection of nitrite, leukocytes, and erythrocytes provided better diagnostic performance than individual detection. Combined detection had higher sensitivity and negative predictive value, while the specificity and positive predictive value were higher for single detection of nitrite. CONCLUSIONS: Present findings demonstrate that the overall performance of the four POCT urine analyzers was sufficient for routine use for screening UTI. A result positive for nitrite is helpful for early diagnosis of UTI, and a result negative for the combination of nitrite, leukocytes, and erythrocytes is helpful for early elimination of UTI.


Assuntos
Infecções Urinárias , Testes Diagnósticos de Rotina , Humanos , Testes Imediatos , Valor Preditivo dos Testes , Sensibilidade e Especificidade , Urinálise , Infecções Urinárias/diagnóstico , Urina
9.
Int J Mol Sci ; 22(19)2021 Sep 29.
Artigo em Inglês | MEDLINE | ID: mdl-34638875

RESUMO

Pluripotent adult stem cells have potential applications in cell therapy and tissue engineering. Urine-derived stem cells (UDSCs) differentiate into various cell types. Here, we attempted to differentiate human UDSCs (hUDSCs) into smooth muscle cells (SMCs) using transforming growth factor-beta 1 (TGF-ß1) and/or PD98059, an extracellular signal-regulated kinase (ERK) inhibitor. Both quantitative polymerase chain reaction (qPCR) and Western blot analysis showed that the expression of messenger ribonucleic acid (mRNA) and proteins for alpha-smooth muscle actin (α-SMA), calponin (CNN1), and smooth muscle myosin heavy chain (SM-MHC), which are specific markers for SMCs, increased on day 9 after differentiation and again on day 14. The differentiated cells from human UDSCs (hUDSCs) with a combination of TGF-ß1 and PD98059 showed the highest expression of SMC marker proteins. Immunocytochemical staining performed to assess the molecular expression revealed CNN and α-SMA colocalizing in the cytoplasm. The cells that differentiated from hUDSCs with a combination of TGF-ß1 and PD98059 showed the strongest expression for CNN1, α-SMA, and SM-MHC. Functional testing of the differentiated cells revealed a stronger contractile capacity for the cells differentiated with a combination of PD98059 and TGF-ß1 than those differentiated with a single factor. These results suggest the combination of PD98059 and TGF-ß1 to be a more effective differentiation method and that differentiated SMCs could be used for restoring the functions of the sphincter muscle or bladder.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Flavonoides/farmacologia , Células Musculares , Células-Tronco , Fator de Crescimento Transformador beta1/farmacologia , Urina/citologia , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Células Musculares/citologia , Células Musculares/metabolismo , Células-Tronco/citologia , Células-Tronco/metabolismo
10.
Int J Mol Sci ; 22(19)2021 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-34638936

RESUMO

Nicotinamide adenine dinucleotide (NAD+) and its reduced form (NADH) are coenzymes employed in hundreds of metabolic reactions. NAD+ also serves as a substrate for enzymes such as sirtuins, poly(ADP-ribose) polymerases (PARPs) and ADP-ribosyl cyclases. Given the pivotal role of NAD(H) in health and disease, studying NAD+ metabolism has become essential to monitor genetic- and/or drug-induced perturbations related to metabolic status and diseases (such as ageing, cancer or obesity), and its possible therapies. Here, we present a strategy based on liquid chromatography-tandem mass spectrometry (LC-MS/MS), for the analysis of the NAD+ metabolome in biological samples. In this method, hydrophilic interaction chromatography (HILIC) was used to separate a total of 18 metabolites belonging to pathways leading to NAD+ biosynthesis, including precursors, intermediates and catabolites. As redox cofactors are known for their instability, a sample preparation procedure was developed to handle a variety of biological matrices: cell models, rodent tissues and biofluids, as well as human biofluids (urine, plasma, serum, whole blood). For clinical applications, quantitative LC-MS/MS for a subset of metabolites was demonstrated for the analysis of the human whole blood of nine volunteers. Using this developed workflow, our methodology allows studying NAD+ biology from mechanistic to clinical applications.


Assuntos
Metaboloma , NAD/biossíntese , Plasma/metabolismo , Soro/metabolismo , Espectrometria de Massas em Tandem/métodos , Urina/fisiologia , Animais , Doadores de Sangue , Cromatografia Líquida/métodos , Células Hep G2 , Humanos , Interações Hidrofóbicas e Hidrofílicas , Metabolômica/métodos , Camundongos , Camundongos Endogâmicos C57BL , Monitorização Fisiológica/métodos , Oxirredução , Projetos Piloto , Plasma/química , Soro/química , Urina/química
11.
Molecules ; 26(17)2021 Aug 27.
Artigo em Inglês | MEDLINE | ID: mdl-34500641

RESUMO

A novel diatomite-supported zeolitic imidazolate framework-8 sorbent (ZIF-8@Dt-COOH) was in situ fabricated and developed for solid-phase extraction of three benzodiazepines (triazolam, midazolam and diazepam) in urine followed by high-performance liquid chromatography. ZIF-8@Dt-COOH was easily prepared by coating ZIF-8 on the surface of Dt-COOH and characterized by Fourier transform infrared spectra, X-ray powder diffractometry and scanning electron microscopy. Compared with bare Dt-COOH, the extraction efficiency of ZIF-8@Dt-COOH for the target was significantly increased from 20.1-39.0% to 100%. Main extraction parameters, including ionic strength and pH of solution, loading volume, washing solution, elution solvent and elution volume, were optimized in detail. Under optimum conditions, the developed method gave linearity of three BZDs in 2-500 ng/mL (r ≥ 0.9995). Limits of detection (S/N = 3), and limits of quantification (S/N = 10) were 0.3-0.4 ng/mL and 1.0-1.3 ng/mL, respectively. In addition, the average recoveries at three spiked levels (5, 10 and 20 ng/mL) varied from 80.0% to 98.7%, with the intra-day and inter-day precisions of 1.4-5.2% and 1.5-8.2%, respectively. The proposed method provided an effective purification performance and gave the enrichment factors of 24.0-29.6. The proposed method was successfully employed for the accurate and sensitive determination of benzodiazepines in urine.


Assuntos
Benzodiazepinas/urina , Terra de Diatomáceas/química , Urina/química , Zeolitas/química , Cromatografia Líquida de Alta Pressão/métodos , Humanos , Limite de Detecção , Extração em Fase Sólida/métodos , Soluções/química , Solventes/química
12.
PLoS One ; 16(9): e0256727, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34492048

RESUMO

Low-iodine diet (LID) is a crucial preparation for radioactive iodine (RAI) treatment or scan in thyroid cancer. The aim of this study is to analyze the influence of thyroid stimulating hormone (TSH) stimulation protocols and other clinical factors on LID adequacy. Thyroid cancer patients who underwent LID for RAI scan or treatment were retrospectively analyzed. Patients were guided to have LID for 2 weeks before RAI administration and urine iodine/creatinine ratio (UICR, µg/g Cr) was measured. TSH stimulation was conducted using either thyroid hormone withdrawal (THW) or recombinant human TSH (rhTSH) injection. Adequacy of LID was classified by UICR as 'excellent (< 50)', 'adequate (50-100)', 'inadequate (101-250)' and 'poor (> 250)'. A total of 1715 UICR measurements from 1054 patients were analyzed. UICR was significantly higher in case of rhTSH use than THW (72.4 ± 48.1 vs. 29.9 ± 45.8 µg/g Cr, P < 0.001). In patients who underwent LID twice using both TSH stimulation protocols alternately, UICR was higher in case of rhTSH than THW regardless of the order of method. Among clinical factors, female, old-age, and the first LID were significant factors to show higher UICR. Although the adequacy of LID was 'adequate' or 'excellent' in most patients, multivariate analysis demonstrated that THW method, male, young age, and prior LID-experience were significant determinants for achieving 'excellent' adequacy of LID. In conclusion, UICR was higher and the proportion of 'excellent' LID adequacy was lower with rhTSH than with THW. UICR was higher also in women, old-age, and LID-naïve patients. Further researches are required to suggest effective methods to reduce body iodine pool in case of rhTSH use and to validate the efficacy of such methods on outcomes of RAI treatment.


Assuntos
Radioisótopos do Iodo/administração & dosagem , Neoplasias da Glândula Tireoide/radioterapia , Tirotropina Alfa/administração & dosagem , Tireotropina/genética , Adulto , Idoso , Dieta , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Neoplasias da Glândula Tireoide/dietoterapia , Neoplasias da Glândula Tireoide/patologia , Neoplasias da Glândula Tireoide/urina , Urina
13.
PLoS One ; 16(9): e0253115, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34534220

RESUMO

BACKGROUND: Evidence indicates that whereas repeated rounds of mass drug administration (MDA) programs have reduced schistosomiasis prevalence to appreciable levels in some communities referred to here as responding villages (R). However, prevalence has remained high or less than anticipated in other areas referred to here as persistent hotspot villages (PHS). Using a cross-sectional quantitative approach, this study investigated the factors associated with sustained high Schistosoma mansoni prevalence in some villages despite repeated high annual treatment coverage in western Kenya. METHOD: Water contact sites selected based on observation of points where people consistently go to collect water, wash clothes, bathe, swim or play (young children), wash cars and harvest sand were mapped using hand-held smart phones on the Commcare platform. Quantitative cross-sectional surveys on behavioral characteristics were conducted using interviewer-based semi-structured questionnaires administered to assess water usage/contact patterns and open defecation. Questionnaires were administered to 15 households per village, 50 pupils per school and 1 head teacher per school. One stool and urine sample was collected from 50 school children aged 9-12 year old and 50 adults from both responding (R) and persistent hotspot (PHS) villages. Stool was analyzed by the Kato-Katz method for eggs of S. mansoni and soil-transmitted helminths. Urine samples were tested using the point-of-care circulating cathodic antigen (POC-CCA) test for detection of S. mansoni antigen. RESULTS: There was higher latrine coverage in R (n = 6) relative to PHS villages (n = 6) with only 33% of schools in the PHS villages meeting the WHO threshold for boy: latrine coverage ratio versus 83.3% in R, while no villages met the girl: latrine ratio requirement. A higher proportion of individuals accessed unprotected water sources for both bathing and drinking (68.5% for children and 89% for adults) in PHS relative to R villages. In addition, frequency of accessing water sources was higher in PHS villages, with swimming being the most frequent activity. As expected based upon selection criteria, both prevalence and intensity of S. mansoni were higher in the PHS relative to R villages (prevalence: 43.7% vs 20.2%; P < 0.001; intensity: 73.8 ± 200.6 vs 22.2 ± 96.0, P < 0.0001), respectively. CONCLUSION: Unprotected water sources and low latrine coverage are contributing factors to PHS for schistosomiasis in western Kenya. Efforts to increase provision of potable water and improvement in latrine infrastructure is recommended to augment control efforts in the PHS areas.


Assuntos
Aparelho Sanitário/parasitologia , Schistosoma mansoni/isolamento & purificação , Esquistossomose/epidemiologia , Solo/parasitologia , Adulto , Animais , Criança , Controle de Doenças Transmissíveis , Estudos Transversais , Fezes/parasitologia , Feminino , Humanos , Quênia/epidemiologia , Masculino , Prevalência , Saúde da População Rural , Esquistossomose/urina , Inquéritos e Questionários , Urina/parasitologia
14.
Sci Rep ; 11(1): 15783, 2021 08 04.
Artigo em Inglês | MEDLINE | ID: mdl-34349163

RESUMO

Youth fountain and aging culprits are usually sought and identified in blood but not urine. Extracellular vesicles (EVs) possess parental cell properties, circulate in blood, CSF and urine, and provide paracrine and remote cell-cell communication messengers. This study investigated whether senescence-associated secretory phenotype (SASP) and immune defense factors in EVs of urine could serve as biomarkers in elderly individuals with and without a comorbidity. Urine samples from young adults and elderly individuals with and without Parkinson disease (PD) were collected and stored at - 80 °C until studies. Urine EVs were separated from a drop-through solution and confirmed by verifying CD9, CD63, CD81 and syntenin expression. The EVs and drop-through solution were subjected to measurement of SASP cytokines and defense factors by Milliplex array assays. Many SASP cytokines and defense factors could be detected in urinary EVs but not urinary solutions. Elderly individuals (age > 60) had significantly higher levels of the SASP-associated factors IL-8, IP-10, GRO, and MCP-1 in EVs (p < 0.05). In contrast, some defense factors, IL-4, MDC and IFNα2 in EVs had significantly lower levels in elderly adults than in young adults (age < 30). Patients with and without PD exhibited a similar SASP profile in EVs but significantly lower levels of IL-10 in the EVs from patients with PD. This study used a simple device to separate urinary EVs from solution for comparisons of SASP and defense mediators between young adults and elders with and without PD. Results from this study indicate that aging signature is present in EVs circulating to urine and the signatures include higher inflammatory mediators and lower defense factors in urinary EVs but not solutions, suggesting a simple method to separate urinary EVs from solutions for searching aging mechanistic biomarkers may make prediction of aging and monitoring of anti-senolytic interventions possible.


Assuntos
Envelhecimento/metabolismo , Vesículas Extracelulares/metabolismo , Doença de Parkinson/metabolismo , Urina/citologia , Adulto , Fatores Etários , Idoso , Biomarcadores/metabolismo , Comunicação Celular , Vesículas Extracelulares/imunologia , Vesículas Extracelulares/fisiologia , Feminino , Humanos , Mediadores da Inflamação/metabolismo , Interleucina-10/metabolismo , Masculino , Pessoa de Meia-Idade , Adulto Jovem
16.
J Chromatogr A ; 1654: 462459, 2021 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-34407470

RESUMO

In current paper, a new advanced modification of fabric phase sorptive extraction is introduced for the first time. This advantageous configuration that integrates the stirring and extraction mechanism into a single sample preparation device was originated by equally considering the beneficial role of the increase of extraction kinetics and more specifically of diffusion on the extraction efficiency of the equilibrium based microextraction techniques and the need for integrating and unite processes for better promotion and implementation of the principles of Green Analytical Chemistry. The resulted magnet integrated fabric phase sorptive extraction (MI-FPSE) device was the spearhead to develop a new analytical methodology for the determination of selected very common endocrine disrupting chemicals as model analytes in human urine by high-performance liquid chromatography-photodiode array analysis. More specifically, the sol-gel Carbowax 20 M coated on hydrophilic cellulose fabric substrate, MI-FPSE device was efficiently employed for the establishment of a new extraction protocol before the chromatographic determination. The sample preparation workflow was methodically optimized in terms of the elution solvent mixture, the volume of the sample, the extraction and the elution time, the stirring speed during the extraction, the ionic strength, and the pH of the sample matrix. The chromatographic separation was performed on a Spherisorb C18 column and a gradient elution program within 14 minutes. Mobile phase consisted of 0.05 ammonium acetate aqueous solution and acetonitrile. The method was validated towards linearity, sensitivity, selectivity, precision, accuracy, and stability. LOD and LOQ ranged between 1.05-1.80 and 3.5-6.0 ng/mL, while %RSD values were found lower than 9.0% in all cases. The method was efficiently applied to the bioanalysis of real samples. All the chosen EDCs were measured at high detection levels. The new MI-FPSE device has demonstrated its performance superiority as a magnet integrated stand-alone extraction device and could be considered as a significant improvement in the field of analytical/bioanalytical sample preparation.


Assuntos
Cromatografia Líquida de Alta Pressão , Disruptores Endócrinos , Urina , Disruptores Endócrinos/isolamento & purificação , Disruptores Endócrinos/urina , Humanos , Interações Hidrofóbicas e Hidrofílicas , Fenômenos Magnéticos , Urina/química
17.
J Chromatogr A ; 1654: 462457, 2021 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-34404016

RESUMO

Signal variation is a common drawback in untargeted metabolomics using liquid chromatography-mass spectrometry (LC-MS), mainly due to the complexity of biological matrices and reduced sample preparation, which results in the accumulation of sample components in the column and the ion source. Here we propose a simple, easy to implement approach to improve data quality in untargeted metabolomics by LC-MS. This approach involves the use of a divert valve to direct the column effluent to waste at the beginning of the chromatographic run and during column cleanup and equilibration, in combination with longer column cleanups in between injections. Our approach was tested using urine samples collected from patients after renal transplantation. Analytical responses were contrasted before and after introducing these modifications by analyzing a batch of untargeted metabolomics data. A significant improvement in peak area repeatability was observed for the quality controls, with relative standard deviations (RSDs) for several metabolites decreasing from ∼60% to ∼10% when our approach was introduced. Similarly, RSDs of peak areas for internal standards improved from ∼40% to ∼10%. Furthermore, calibrant solutions were more consistent after introducing these modifications when comparing peak areas of solutions injected at the beginning and the end of each analytical sequence. Therefore, we recommend the use of a divert valve and extended column cleanup as a powerful strategy to improve data quality in untargeted metabolomics, especially for very complex types of samples where minimum sample preparation is required, such as in this untargeted metabolomics study with urine from renal transplanted patients.


Assuntos
Cromatografia Líquida , Confiabilidade dos Dados , Espectrometria de Massas , Metabolômica , Urinálise , Humanos , Urinálise/métodos , Urinálise/normas , Urina/química
18.
Ann Clin Lab Sci ; 51(4): 562-569, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34452897

RESUMO

OBJECTIVE: The aim of this study was to evaluate the consistency and accuracy of all the parameters of the urine samples detected by two automated urine sediment analyzers from Sysmex Corporation. METHODS: Two automated analyzers and manual microscopy examined 1,059 urine samples. The sensitivity, specificity, positive predictive value, and negative predictive value were evaluated. The consistency of all the parameters was tested. The influencing factors of false positive and false negative samples were analyzed and compared. RESULTS: All the parameters had good specificity, negative predictive value, and coincidence rate (83.95%-99.61%). The RBC, WBC, and X'TAL analyzed by UF-5000 and UF-1000i exhibited good agreement (Kappa=0.597-0.784) with those by manual microscopy. The overall concordance rates of RBC and WBC were good (RBC: r=0.9842, CCC=0.9693; WBC: r=0.9955, CCC=0.9711). Among the influencing factors, mucus filament accounted for a large proportion, which mainly affected the detection of CAST. Concurrently, the false-positive factors of EC detection were reduced, and CAST did not affect the detection of EC. CONCLUSION: The parameters of the two instruments tested have shown high accuracy, consistency, coincidence rate, and low negative predictive value for RBC and WBC, which has ensured that UF-5000 and UF-1000i meet the clinical requirements for urine tests for disease screening. For the samples with poor consistency and false-positive factors, a conventional microscopic examination should be applied to verify the accuracy of the instrument detection.


Assuntos
Células Epiteliais/citologia , Citometria de Fluxo/instrumentação , Leucócitos/citologia , Urinálise/instrumentação , Urina/citologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Adulto Jovem
19.
Angew Chem Int Ed Engl ; 60(43): 23232-23240, 2021 10 18.
Artigo em Inglês | MEDLINE | ID: mdl-34339587

RESUMO

The microbiome has a fundamental impact on the human host's physiology through the production of highly reactive compounds that can lead to disease development. One class of such compounds are carbonyl-containing metabolites, which are involved in diverse biochemical processes. Mass spectrometry is the method of choice for analysis of metabolites but carbonyls are analytically challenging. Herein, we have developed a new chemical biology tool using chemoselective modification to overcome analytical limitations. Two isotopic probes allow for the simultaneous and semi-quantitative analysis at the femtomole level as well as qualitative analysis at attomole quantities that allows for detection of more than 200 metabolites in human fecal, urine and plasma samples. This comprehensive mass spectrometric analysis enhances the scope of metabolomics-driven biomarker discovery. We anticipate that our chemical biology tool will be of general use in metabolomics analysis to obtain a better understanding of microbial interactions with the human host and disease development.


Assuntos
Acetaldeído/análise , Acetona/análise , Aldeídos/análise , Butanonas/análise , Di-Hidroxiacetona/análise , Metabolômica/métodos , Acetaldeído/sangue , Acetaldeído/química , Acetaldeído/urina , Acetamidas/química , Acetona/sangue , Acetona/química , Acetona/urina , Aldeídos/sangue , Aldeídos/química , Aldeídos/urina , Butanonas/sangue , Butanonas/química , Butanonas/urina , Carbono/química , Isótopos de Carbono/química , Di-Hidroxiacetona/sangue , Di-Hidroxiacetona/química , Di-Hidroxiacetona/urina , Fezes/química , Microbioma Gastrointestinal , Humanos , Indicadores e Reagentes/química , Limite de Detecção , Urina/química
20.
PLoS Med ; 18(8): e1003732, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-34464379

RESUMO

BACKGROUND: The standard of care treatment for muscle-invasive bladder cancer (MIBC) is radical cystectomy, which is typically preceded by neoadjuvant chemotherapy. However, the inability to assess minimal residual disease (MRD) noninvasively limits our ability to offer bladder-sparing treatment. Here, we sought to develop a liquid biopsy solution via urine tumor DNA (utDNA) analysis. METHODS AND FINDINGS: We applied urine Cancer Personalized Profiling by Deep Sequencing (uCAPP-Seq), a targeted next-generation sequencing (NGS) method for detecting utDNA, to urine cell-free DNA (cfDNA) samples acquired between April 2019 and November 2020 on the day of curative-intent radical cystectomy from 42 patients with localized bladder cancer. The average age of patients was 69 years (range: 50 to 86), of whom 76% (32/42) were male, 64% (27/42) were smokers, and 76% (32/42) had a confirmed diagnosis of MIBC. Among MIBC patients, 59% (19/32) received neoadjuvant chemotherapy. utDNA variant calling was performed noninvasively without prior sequencing of tumor tissue. The overall utDNA level for each patient was represented by the non-silent mutation with the highest variant allele fraction after removing germline variants. Urine was similarly analyzed from 15 healthy adults. utDNA analysis revealed a median utDNA level of 0% in healthy adults and 2.4% in bladder cancer patients. When patients were classified as those who had residual disease detected in their surgical sample (n = 16) compared to those who achieved a pathologic complete response (pCR; n = 26), median utDNA levels were 4.3% vs. 0%, respectively (p = 0.002). Using an optimal utDNA threshold to define MRD detection, positive utDNA MRD detection was highly correlated with the absence of pCR (p < 0.001) with a sensitivity of 81% and specificity of 81%. Leave-one-out cross-validation applied to the prediction of pathologic response based on utDNA MRD detection in our cohort yielded a highly significant accuracy of 81% (p = 0.007). Moreover, utDNA MRD-positive patients exhibited significantly worse progression-free survival (PFS; HR = 7.4; 95% CI: 1.4-38.9; p = 0.02) compared to utDNA MRD-negative patients. Concordance between urine- and tumor-derived mutations, determined in 5 MIBC patients, was 85%. Tumor mutational burden (TMB) in utDNA MRD-positive patients was inferred from the number of non-silent mutations detected in urine cfDNA by applying a linear relationship derived from The Cancer Genome Atlas (TCGA) whole exome sequencing of 409 MIBC tumors. We suggest that about 58% of these patients with high inferred TMB might have been candidates for treatment with early immune checkpoint blockade. Study limitations included an analysis restricted only to single-nucleotide variants (SNVs), survival differences diminished by surgery, and a low number of DNA damage response (DRR) mutations detected after neoadjuvant chemotherapy at the MRD time point. CONCLUSIONS: utDNA MRD detection prior to curative-intent radical cystectomy for bladder cancer correlated significantly with pathologic response, which may help select patients for bladder-sparing treatment. utDNA MRD detection also correlated significantly with PFS. Furthermore, utDNA can be used to noninvasively infer TMB, which could facilitate personalized immunotherapy for bladder cancer in the future.


Assuntos
Biomarcadores Tumorais/análise , Cistectomia/estatística & dados numéricos , DNA de Neoplasias/análise , Neoplasia Residual/diagnóstico , Neoplasias da Bexiga Urinária/diagnóstico , Urina/química , Idoso , Idoso de 80 Anos ou mais , Estudos de Coortes , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Missouri , Invasividade Neoplásica/patologia , Neoplasia Residual/etiologia , Intervalo Livre de Progressão , Neoplasias da Bexiga Urinária/etiologia
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