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1.
Nat Commun ; 11(1): 5070, 2020 10 08.
Artigo em Inglês | MEDLINE | ID: mdl-33033260

RESUMO

The evolutionary progression from primary to metastatic prostate cancer is largely uncharted, and the implications for liquid biopsy are unexplored. We infer detailed reconstructions of tumor phylogenies in ten prostate cancer patients with fatal disease, and investigate them in conjunction with histopathology and tumor DNA extracted from blood and cerebrospinal fluid. Substantial evolution occurs within the prostate, resulting in branching into multiple spatially intermixed lineages. One dominant lineage emerges that initiates and drives systemic metastasis, where polyclonal seeding between sites is common. Routes to metastasis differ between patients, and likely genetic drivers of metastasis distinguish the metastatic lineage from the lineage that remains confined to the prostate within each patient. Body fluids capture features of the dominant lineage, and subclonal expansions that occur in the metastatic phase are non-uniformly represented. Cerebrospinal fluid analysis reveals lineages not detected in blood-borne DNA, suggesting possible clinical utility.


Assuntos
Linhagem da Célula , Biópsia Líquida , Neoplasias da Próstata/patologia , Líquidos Corporais/metabolismo , Cromossomos Humanos Par 8/genética , Células Clonais , Variações do Número de Cópias de DNA/genética , DNA de Neoplasias/genética , Loci Gênicos , Humanos , Masculino , Pessoa de Meia-Idade , Metástase Neoplásica , Filogenia
2.
Medicine (Baltimore) ; 99(31): e21460, 2020 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-32756167

RESUMO

Volume status is a key parameter for cardiovascular-related mortality in dialysis patients. Although N-terminal pro-B-type natriuretic peptide (NT-proBNP), myeloperoxidase, copeptin, and pro-adrenomedullin have been reported as volume markers, the relationship between body fluid status and volume markers in dialysis patients is uncertain. Therefore, we investigated the utility of volume status biomarkers based on body composition monitor (BCM) analyses.We enrolled pre-dialysis, hemodialysis (HD), and peritoneal dialysis (PD) patients and age- and gender-matched healthy Korean individuals (N = 80). BCM and transthoracic echocardiography were performed and NT-proBNP, myeloperoxidase, copeptin, and pro-adrenomedullin concentrations were measured. Relative hydration status (ΔHS, %) was defined in terms of the hydration status-to-extracellular water ratio with a cutoff of 15%, and hyperhydrated status was defined as ΔHS > 15%.Although there were no significant differences in total body water, extracellular water, or intracellular water among groups, mean amount of volume overload and hyperhydrated status were significantly higher in HD and PD patients compared with control and pre-dialysis patients. Mean amount of volume overload and hyperhydrated status were also significantly associated with higher NT-proBNP and pro-adrenomedullin levels in HD and PD patients, although not with myeloperoxidase or copeptin levels. Furthermore, they were significantly associated with cardiac markers (left ventricular mass index, ejection fraction, and left atrial diameter) in HD and PD patients compared with those in the control and pre-dialysis groups.On the basis of increased plasma NT-proBNP and pro-adrenomedullin concentrations, we might be able to make predictions regarding the volume overload status of dialysis patients, and thereby reduce cardiovascular-related mortality through appropriate early volume control.


Assuntos
Biomarcadores/sangue , Líquidos Corporais/metabolismo , Doenças Cardiovasculares/mortalidade , Disfunção Ventricular Esquerda/fisiopatologia , Adrenomedulina/sangue , Adulto , Composição Corporal/fisiologia , Doenças Cardiovasculares/diagnóstico por imagem , Estudos de Casos e Controles , Diálise/métodos , Diálise/tendências , Ecocardiografia/métodos , Feminino , Glicopeptídeos/sangue , Humanos , Masculino , Pessoa de Meia-Idade , Peptídeo Natriurético Encefálico/sangue , Fragmentos de Peptídeos/sangue , Diálise Peritoneal/estatística & dados numéricos , Peroxidase/sangue , Precursores de Proteínas/sangue , Diálise Renal/estatística & dados numéricos , República da Coreia/epidemiologia , Volume Sistólico , Resultado do Tratamento , Disfunção Ventricular Esquerda/diagnóstico por imagem
3.
Protein Cell ; 11(10): 707-722, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32519302

RESUMO

The 2019 novel coronavirus disease (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has occurred in China and around the world. SARS-CoV-2-infected patients with severe pneumonia rapidly develop acute respiratory distress syndrome (ARDS) and die of multiple organ failure. Despite advances in supportive care approaches, ARDS is still associated with high mortality and morbidity. Mesenchymal stem cell (MSC)-based therapy may be an potential alternative strategy for treating ARDS by targeting the various pathophysiological events of ARDS. By releasing a variety of paracrine factors and extracellular vesicles, MSC can exert anti-inflammatory, anti-apoptotic, anti-microbial, and pro-angiogenic effects, promote bacterial and alveolar fluid clearance, disrupt the pulmonary endothelial and epithelial cell damage, eventually avoiding the lung and distal organ injuries to rescue patients with ARDS. An increasing number of experimental animal studies and early clinical studies verify the safety and efficacy of MSC therapy in ARDS. Since low cell engraftment and survival in lung limit MSC therapeutic potentials, several strategies have been developed to enhance their engraftment in the lung and their intrinsic, therapeutic properties. Here, we provide a comprehensive review of the mechanisms and optimization of MSC therapy in ARDS and highlighted the potentials and possible barriers of MSC therapy for COVID-19 patients with ARDS.


Assuntos
Betacoronavirus , Infecções por Coronavirus/complicações , Transplante de Células-Tronco Mesenquimais , Pandemias , Pneumonia Viral/complicações , Síndrome do Desconforto Respiratório do Adulto/terapia , Transferência Adotiva , Células Epiteliais Alveolares/patologia , Animais , Apoptose , Líquidos Corporais/metabolismo , Linfócitos T CD4-Positivos/imunologia , Ensaios Clínicos como Assunto , Coinfecção/prevenção & controle , Coinfecção/terapia , Infecções por Coronavirus/imunologia , Modelos Animais de Doenças , Células Endoteliais/patologia , Oxigenação por Membrana Extracorpórea , Terapia Genética/métodos , Vetores Genéticos/administração & dosagem , Vetores Genéticos/uso terapêutico , Humanos , Imunidade Inata , Mediadores da Inflamação/metabolismo , Pulmão/patologia , Pulmão/fisiopatologia , Transplante de Células-Tronco Mesenquimais/métodos , Células-Tronco Mesenquimais/fisiologia , Insuficiência de Múltiplos Órgãos/etiologia , Insuficiência de Múltiplos Órgãos/prevenção & controle , Pneumonia Viral/imunologia , Síndrome do Desconforto Respiratório do Adulto/imunologia , Síndrome do Desconforto Respiratório do Adulto/patologia , Pesquisa Médica Translacional
4.
J Alzheimers Dis ; 76(1): 27-31, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32568212

RESUMO

The coronavirus disease 2019 (COVID-19) pandemic led to an abrupt halt of many Alzheimer's disease (AD) research studies at sites spanning the world. This is especially true for studies requiring in-person contact, such as studies collecting biofluids. Since COVID-19 is likely to remain a threat for an extended period, the resumption of fluid biomarker studies requires the development and implementation of procedures that minimize the risk of in-person visits to participants, staff, and individuals handling the biofluid samples. Some issues to consider include structuring the visit workflow to minimize contacts and promote social distancing; screening and/or testing participants and staff for COVID-19; wearing masks and performing hand hygiene; and precautions for handling, storing, and analyzing biofluids. AD fluid biomarker research remains a vitally important public health priority and resuming studies requires appropriate safety procedures to protect research participants and staff.


Assuntos
Doença de Alzheimer/metabolismo , Betacoronavirus , Infecções por Coronavirus/metabolismo , Pessoal de Saúde/tendências , Segurança do Paciente , Equipamento de Proteção Individual , Pneumonia Viral/metabolismo , Doença de Alzheimer/diagnóstico , Biomarcadores/metabolismo , Líquidos Corporais/metabolismo , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/transmissão , Humanos , Pandemias , Equipamento de Proteção Individual/tendências , Pneumonia Viral/diagnóstico , Pneumonia Viral/transmissão
5.
Yakugaku Zasshi ; 140(5): 617-624, 2020.
Artigo em Japonês | MEDLINE | ID: mdl-32378661

RESUMO

Pancreatic cancer is the fourth-leading cause of death from cancer in Japan, after lung, colorectal, and stomach cancers and has the lowest survival among these tumors, because of not only no symptoms, no screening tool and no biomarkers but also high rates of recurrence and metastasis. In addition, pancreatic cancer has excessive stroma which serves as a severe biological barrier for anticancer drug delivery and successful treatment. Therefore, there are many challenges for drug delivery systems for the treatment of pancreatic cancer. Recently, we developed self-assembly PEGylation retaining activity (SPRA) technology, which comprises a reversible pegylated protein complex without loss of bioactivity. SPRA technology is based on a host-guest interaction between PEGylated ß-cyclodextrin and adamantane-appended protein. In this review, first pancreatic cancer is introduced, second, principle drug delivery systems for the treatment of pancreatic cancer are described, and third the concept of SPRA technology as well as examples of SPRA proteins, especially focusing on the potential of SPRA-bromelain for treatment of pancreatic cancer, are introduced.


Assuntos
Antineoplásicos/administração & dosagem , Líquidos Corporais/metabolismo , Sistemas de Liberação de Medicamentos/métodos , Substâncias Macromoleculares , Neoplasias Pancreáticas/tratamento farmacológico , Neoplasias Pancreáticas/metabolismo , Adamantano , Bromelaínas , Humanos , Polietilenoglicóis , Pressão , beta-Ciclodextrinas
6.
J Vis Exp ; (159)2020 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-32420991

RESUMO

In situ hybridization (ISH) is a very informative technique to present cellular distribution patterns of specific genes (e.g., mRNA and ncRNA) in tissues. The sipunculid worm Sipunculus nudus is a crucial fishery resource as it has high nutritional and medicinal values. Currently, the research on the molecular biology of Sipunculus nudus is still in its infancy. The purpose of this article is to develop a sensitive method for localizing specific mRNA in Sipunculus nudus coelomic fluid. The protocol includes detailed steps of ISH, including digoxigenin-labeled antisense and sense riboprobe preparation, coelomic fluid collection and section preparation, specific riboprobe hybridization, antibody incubation, coloration and post-coloration treatments. The representative results obtained from a successful experiment using this method are demonstrated. The protocol should be applicable to other Sipuncula species as well.


Assuntos
Líquidos Corporais/metabolismo , Hibridização In Situ , Poliquetos/genética , Animais , Reação em Cadeia da Polimerase , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
8.
J Med Life ; 13(1): 21-25, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32341696

RESUMO

Immunopathogenesis of inflammatory and dystrophic diseases of the tissues of the oral cavity is characterized by cellular and humoral factors of specific and nonspecific resistance, the functioning of which is determined by the overall somatic state. This study aimed to study the features of protective mechanisms of the oral cavity due to orthodontic pathology, pathology of periodontal tissues, and odontogenic inflammatory process in children with diffuse nontoxic goiter. Eighty children with diffuse nontoxic goiter aged 12-15 years with different dental status were examined. Evaluation of local immunity of the oral cavity was carried out by determining the content of sIgA, IgA, IgG, lysozyme activity, and levels of IL-1ß, IL-4 by enzyme immunoassay. Immunological studies have shown that in children with diffuse nontoxic goiter, the activity of lysozyme in the oral fluid is decreased. The level of sIgА is also reduced by about 20%. Besides, there is an increase in the content of IgG and a growing trend in the level of IgА. The content of IL-1ß and IL-4 in such children fluctuates more compared to somatically healthy children. In conclusion, a violation of the local protective mechanisms of the oral cavity is observed in children with diffuse nontoxic goiter. Also, the increase in the severity of dental pathology leads to increased tension of local protective and compensatory reactions.


Assuntos
Bócio/patologia , Boca/patologia , Adolescente , Líquidos Corporais/metabolismo , Criança , Humanos , Imunoglobulinas/metabolismo , Interleucina-1beta/metabolismo , Interleucina-4/metabolismo , Masculino
9.
PLoS One ; 15(4): e0229976, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32275679

RESUMO

Small extracellular vesicles (sEV) are nano-sized (40-150 nm), membrane-encapsulated vesicles that are released by essentially all cells into the extracellular space and function as intercellular signaling vectors through the horizontal transfer of biologic molecules, including microRNA (miRNA) and other small non-coding RNA (ncRNA), that can alter the phenotype of recipient cells. sEV are present in essentially all extracellular biofluids, including serum, urine and saliva, and offer a new avenue for discovery and development of novel biomarkers of various disease states and exposures. The objective of this study was to systematically interrogate similarities and differences between sEV ncRNA derived from saliva, serum and urine, as well as cell-free small ncRNA (cf-ncRNA) from serum. Saliva, urine and serum were concomitantly collected from 4 healthy donors to mitigate potential bias that can stem from interpersonal and temporal variability. sEV were isolated from each respective biofluid, along with cf-RNA from serum. sEV were isolated from the respective biofluids via differential ultracentrifugation with a 30% sucrose cushion to minimize protein contamination. Small RNA-sequencing was performed on each sample, and cluster analysis was performed based on ncRNA profiles. While some similarities existed in terms of sEV ncRNA cargo across biofluids, there are also notable differences in ncRNA class and ncRNA secretion, with sEV in each biofluid bearing a unique ncRNA profile, including major differences in composition by ncRNA class. We conclude that sEV ncRNA cargo varies according to biofluid, so thus should be carefully selected and interpreted when designing or contrasting translational or epidemiological studies.


Assuntos
Biomarcadores/análise , Vesículas Extracelulares/metabolismo , MicroRNAs/análise , Pequeno RNA não Traduzido/análise , Adulto , Idoso , Biomarcadores/sangue , Biomarcadores/urina , Líquidos Corporais/metabolismo , Testes Diagnósticos de Rotina/métodos , Feminino , Humanos , Masculino , MicroRNAs/sangue , MicroRNAs/urina , Pessoa de Meia-Idade , Pequeno RNA não Traduzido/sangue , Pequeno RNA não Traduzido/urina , Saliva/metabolismo , Análise de Sequência de RNA , Ultracentrifugação
10.
J Dairy Sci ; 103(6): 5047-5060, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32278566

RESUMO

Ruminants are born with an undeveloped physical, metabolic, and microbial rumen. Rumen development is limited under artificial rearing systems when newborn animals are separated from the dam, fed on milk replacer, and weaned at an early age. This study aims to evaluate the effects of early-life inoculation of young ruminants with rumen fluid from adult animals. Eighty newborn goat kids were randomly allocated to 1 of 4 experimental treatments and inoculated daily from d 1 to wk 11 with autoclaved rumen fluid (AUT), fresh rumen fluid obtained from adult goats fed either a forage diet (RFF) or concentrate-rich diet (RFC), or absence of inoculation (CTL). Goat kids were artificially reared with ad libitum access to milk replacer, starter concentrate, and forage hay. Blood was sampled weekly and rumen microbial fermentation was monitored at 5 (preweaning), 7 (weaning), and 9 wk of age (postweaning). Results indicated that inoculation with fresh rumen fluid accelerated the rumen microbial and fermentative development before weaning. As a result, RFC and RFF animals had higher solid feed intake (+73%), rumen concentrations of ammonia-N (+26%), total volatile fatty acids (+46%), butyrate (+50%), and plasma ß-hydroxybutyrate (+48%), and lower milk intake (-6%) than CTL and AUT animals at wk 5. Inoculation with fresh inoculum also promoted early rumen colonization by a complex and abundant protozoal community, whereas CTL animals remained protozoa free. Although all kids experienced moderate growth retardation during 1 wk after weaning, inoculation with fresh rumen fluid favored the weaning process, leading to 2.2 times higher weight gain than CTL and AUT animals during wk 8. Some of these advantages were retained during the postweaning period and RFF and RFC animals showed higher forage intake (up to +44%) than CTL and AUT animals with no detrimental effects on feed digestibility or stress levels. The superior microbial load of RFC compared with RFF inoculum tended to provide further improvements in terms of forage intake, plasma ß-hydroxybutyrate, and rumen protozoa, whereas AUT inoculation provided minor (if any) advantages with respect to CTL animals. Although no differences were noted on animal growth, this study suggests that early life inoculation of goat kids with rumen microbiota can represent an effective strategy to accelerate the rumen development, facilitating a smooth transition from milk to solid feed and to the potential implementation of early weaning strategies.


Assuntos
Líquidos Corporais , Cabras , Rúmen , Desmame , Ácido 3-Hidroxibutírico/sangue , Ração Animal , Animais , Animais Recém-Nascidos , Líquidos Corporais/metabolismo , Butiratos/metabolismo , Dieta/veterinária , Ácidos Graxos Voláteis/metabolismo , Fermentação , Cabras/metabolismo , Microbiota , Leite/metabolismo , Rúmen/metabolismo , Ganho de Peso
11.
Cancer Cytopathol ; 128(6): 384-391, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32163239

RESUMO

BACKGROUND: Body fluid cytology (BFC) is an important tool in the diagnosis and staging of malignancy and is aided by the judicious use of immunohistochemistry (IHC). The aim of this study was to determine the usage rates of IHC stains in BFC, their type and indications, and their diagnostic impact. We also attempted to estimate the optimal rate of IHC use in BFC by comparing the entire laboratory's and each individual cytopathologist's IHC use rates with their respective indeterminate and malignant diagnosis rates. METHODS: We conducted a retrospective study of IHC stain use in BFC during a 5.5-year interval (2013-2018) and determined the laboratory's and each individual cytopathologist's IHC usage patterns according to the final diagnosis, site, and indications for their use. RESULTS: A total of 477 out of 4144 (11.5%) BFC cases had 2128 individual immunostains performed, with an average of 4.5 immunostains per case. Individual cytopathologists used IHC stains on 6.7% to 22% of their BFC cases. Pathologists with higher rates of IHC stain use than the laboratory's mean were less experienced and had higher rates of indeterminate but not of malignant diagnoses. The most common indication for the use of IHC stains was differentiating mesothelial from malignant cells. MOC31, calretinin, Ber-EP4, CD68, and D2-40 were the most commonly used of the 67 different IHC stains used in BFC. CONCLUSIONS: The laboratory's mean may represent the optimal IHC use rate, as higher IHC use rates did not lead to more diagnostic certainty or higher pickup rates of malignant cells.


Assuntos
Biomarcadores Tumorais/metabolismo , Líquidos Corporais/metabolismo , Citodiagnóstico/métodos , Imuno-Histoquímica/métodos , Neoplasias/diagnóstico , Líquidos Corporais/citologia , Diagnóstico Diferencial , Humanos , Neoplasias/metabolismo , Patologistas/normas , Patologistas/estatística & dados numéricos , Patologia Clínica/métodos , Patologia Clínica/normas , Patologia Clínica/estatística & dados numéricos , Padrões de Prática Médica/normas , Padrões de Prática Médica/estatística & dados numéricos , Reprodutibilidade dos Testes , Estudos Retrospectivos , Sensibilidade e Especificidade , Coloração e Rotulagem/métodos
12.
Sci Rep ; 10(1): 3341, 2020 02 24.
Artigo em Inglês | MEDLINE | ID: mdl-32094409

RESUMO

High-throughput sequencing technologies could improve diagnosis and classification of TBI subgroups. Because recent studies showed that circulating microRNAs (miRNAs) may serve as noninvasive markers of TBI, we performed miRNA-seq to study TBI-induced changes in rat hippocampal miRNAs up to one year post-injury. We used miRNA PCR arrays to interrogate differences in serum miRNAs using two rat models of TBI (controlled cortical impact [CCI] and fluid percussion injury [FPI]). The translational potential of our results was evaluated by miRNA-seq analysis of human control and TBI (acute and chronic) serum samples. Bioinformatic analyses were performed using Ingenuity Pathway Analysis, miRDB, and Qlucore Omics Explorer. Rat miRNA profiles identified TBI across all acute and chronic intervals. Rat CCI and FPI displayed distinct serum miRNA profiles. Human miRNA profiles identified TBI across all acute and chronic time points and, at 24 hours, discriminated between focal and diffuse injuries. In both species, predicted gene targets of differentially expressed miRNAs are involved in neuroplasticity, immune function and neurorestoration. Chronically dysregulated miRNAs (miR-451a, miR-30d-5p, miR-145-5p, miR-204-5p) are linked to psychiatric and neurodegenerative disorders. These data suggest that circulating miRNAs in biofluids can be used as "molecular fingerprints" to identify acute, chronic, focal or diffuse TBI and potentially, presence of neurodegenerative sequelae.


Assuntos
Líquidos Corporais/metabolismo , Lesões Encefálicas Traumáticas/genética , Hipocampo/metabolismo , MicroRNAs/genética , Análise de Sequência de RNA , Doença Aguda , Adulto , Animais , Doença Crônica , Humanos , MicroRNAs/metabolismo , Pessoa de Meia-Idade , Análise de Componente Principal , Ratos , Transdução de Sinais/genética
13.
Sci Rep ; 10(1): 2783, 2020 02 17.
Artigo em Inglês | MEDLINE | ID: mdl-32066796

RESUMO

Treatment of uveitis is complicated because of its multiple aetiologies and elevation of various inflammatory mediators. To determine the mediators that are elevated in the vitreous humor according to the aetiology of the uveitis, we examined the concentrations of 21 inflammatory cytokines, 7 chemokines, and 5 colony-stimulating/growth factors in vitreous samples from 57 eyes with uveitis associated with intraocular lymphoma (IOL, n = 13), sarcoidosis (n = 15), acute retinal necrosis (ARN, n = 13), or bacterial endophthalmitis (BE, n = 16). Samples from eyes with idiopathic epiretinal membrane (n = 15), which is not associated with uveitis, were examined as controls. Heat map analysis demonstrated that the patterns of inflammatory mediators in the vitreous humor in eyes with uveitis were disease-specific. Pairwise comparisons between the 5 diseases showed specific elevation of interferon-α2 in ARN and interleukin (IL)-6, IL-17A, and granulocyte-colony stimulating factor in BE. Pairwise comparisons between IOL, ARN, and BE revealed that levels of IL-10 in IOL, RANTES (regulated on activation, normal T cell expressed and secreted) in ARN, and IL-22 in BE were significantly higher than those in the other 2 types of uveitis. These mediators are likely to be involved in the immunopathology of specific types of uveitis and may be useful biomarkers.


Assuntos
Biomarcadores/metabolismo , Inflamação/metabolismo , Uveíte/metabolismo , Corpo Vítreo/metabolismo , Idoso , Líquidos Corporais/metabolismo , Endoftalmite/complicações , Endoftalmite/epidemiologia , Endoftalmite/patologia , Membrana Epirretiniana/patologia , Olho/metabolismo , Olho/patologia , Feminino , Humanos , Inflamação/complicações , Inflamação/patologia , Interleucina-6/metabolismo , Linfoma Intraocular/complicações , Linfoma Intraocular/epidemiologia , Linfoma Intraocular/patologia , Masculino , Pessoa de Meia-Idade , Síndrome de Necrose Retiniana Aguda/complicações , Síndrome de Necrose Retiniana Aguda/epidemiologia , Síndrome de Necrose Retiniana Aguda/patologia , Sarcoidose/complicações , Sarcoidose/epidemiologia , Sarcoidose/patologia , Uveíte/complicações , Uveíte/patologia , Corpo Vítreo/patologia
14.
Ann Biol Clin (Paris) ; 78(1): 93-107, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-32108587

RESUMO

The measurement performance of 13 biochemistry parameters (CEA, CA 19-9, amylase, lipase, sodium, potassium, chloride, creatinine, glucose, protein, albumin, LDH, triglycerides) was tested in a panel of biological fluids other than blood and urine (peritoneal, pleural, pancreatic fluids ...). Our protocol, based on a risk analysis, allowed us to justify our choices and compare the performance obtained with those of the serum or plasma matrix already validated. Thus, the coefficients of variation obtained in body fluids are comparable. The assessment of accuracy (spiking and dilution tests) shows the absence of bias, which is consistent with the absence of matrix effect. The linearity studied by dilution tests shows that the upper limits of the measurement interval communicated by the supplier are applicable to body fluids. The absence of contamination and stability have been also confirmed. All analytes are stable for 3 days at room temperature, 7 days between 2 and 8̊C, and 6 months at -20̊C; except LDH and lipase. For most analytes, at least one interference (hemolysis, icterus, lipemia) was found. Finally, a bibliographical study, confronted with the experience of prescribers, led us to define optimal thresholds to help interpret patients' results. In conclusion, this work has allowed us to validate analytical methods for body fluids testing after relying on their comparability to the blood matrix. We have also been able to adapt our practices and finally be accredited according to the standard NF IN ISO 15189.


Assuntos
Biomarcadores/análise , Líquidos Corporais/química , Técnicas de Laboratório Clínico , Albuminas/análise , Albuminas/metabolismo , Amilases/análise , Amilases/metabolismo , Biomarcadores/metabolismo , Líquidos Corporais/metabolismo , Antígeno CA-19-9/análise , Antígeno CA-19-9/metabolismo , Antígeno Carcinoembrionário/análise , Antígeno Carcinoembrionário/metabolismo , Cloretos/análise , Cloretos/metabolismo , Técnicas de Laboratório Clínico/métodos , Técnicas de Laboratório Clínico/normas , Creatinina/análise , Creatinina/metabolismo , Glucose/análise , Glucose/metabolismo , Humanos , L-Lactato Desidrogenase/análise , L-Lactato Desidrogenase/metabolismo , Lipase/análise , Lipase/metabolismo , Potássio/análise , Potássio/metabolismo , Proteínas/análise , Proteínas/metabolismo , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Sódio/análise , Sódio/metabolismo , Temperatura , Triglicerídeos/análise , Triglicerídeos/metabolismo
15.
Physiol Rep ; 8(2): e14360, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31994353

RESUMO

Most of the filtered glucose is reabsorbed in the early proximal tubule by the sodium-glucose cotransporter SGLT2. The glycosuric effect of the SGLT2 inhibitor ipragliflozin is linked to a diuretic and natriuretic effect that activates compensatory increases in fluid and food intake to stabilize body fluid volume (BFV). However, the compensatory mechanisms that are activated on the level of renal tubules remain unclear. Type 2 diabetic Goto-Kakizaki (GK) rats were treated with vehicle or 0.01% (in diet) ipragliflozin with free access to fluid and food. After 8 weeks, GK rats were placed in metabolic cages for 24-hr. Ipragliflozin decreased body weight, serum glucose and systolic blood pressure, and increased fluid and food intake, urinary glucose and Na+ excretion, urine volume, and renal osmolar clearance, as well as urine vasopressin and solute-free water reabsorption (TcH2O). BFV, measured by bioimpedance spectroscopy, and fluid balance were similar among the two groups. Urine vasopressin in ipragliflozin-treated rats was negatively and positively associated with fluid balance and TcH2O, respectively. Ipragliflozin increased the renal membrane protein expression of SGLT2, aquaporin (AQP) 2 phosphorylated at Ser269 and vasopressin V2 receptor. The expression of SGLT1, GLUT2, AQP1, and AQP2 was similar between the groups. In conclusion, the SGLT2 inhibitor ipragliflozin induced a sustained glucosuria, diuresis, and natriuresis, with compensatory increases in fluid intake and vasopressin-induced TcH2O in proportion to the reduced fluid balance to maintain BFV. These results indicate that the osmotic diuresis induced by SGLT2 inhibition stimulates compensatory fluid intake and renal water reabsorption to maintain BFV.


Assuntos
Líquidos Corporais/metabolismo , Diurese/fisiologia , Osmose/fisiologia , Reabsorção Renal/fisiologia , Inibidores do Transportador 2 de Sódio-Glicose/farmacologia , Vasopressinas/urina , Água/metabolismo , Animais , Compartimentos de Líquidos Corporais/efeitos dos fármacos , Compartimentos de Líquidos Corporais/metabolismo , Líquidos Corporais/efeitos dos fármacos , Diurese/efeitos dos fármacos , Diuréticos Osmóticos/farmacologia , Glucosídeos/farmacologia , Osmose/efeitos dos fármacos , Ratos , Reabsorção Renal/efeitos dos fármacos , Tiofenos/farmacologia
16.
Pharm Res ; 37(3): 42, 2020 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-31989335

RESUMO

PURPOSE: The design of biorelevant conditions for in vitro evaluation of orally administered drug products is contingent on obtaining accurate values for physiologically relevant parameters such as pH, buffer capacity and bile salt concentrations in upper gastrointestinal fluids. METHODS: The impact of sample handling on the measurement of pH and buffer capacity of aspirates from the upper gastrointestinal tract was evaluated, with a focus on centrifugation and freeze-thaw cycling as factors that can influence results. Since bicarbonate is a key buffer system in the fasted state and is used to represent conditions in the upper intestine in vitro, variations on sample handling were also investigated for bicarbonate-based buffers prepared in the laboratory. RESULTS: Centrifugation and freezing significantly increase pH and decrease buffer capacity in samples obtained by aspiration from the upper gastrointestinal tract in the fasted state and in bicarbonate buffers prepared in vitro. Comparison of data suggested that the buffer system in the small intestine does not derive exclusively from bicarbonates. CONCLUSIONS: Measurement of both pH and buffer capacity immediately after aspiration are strongly recommended as "best practice" and should be adopted as the standard procedure for measuring pH and buffer capacity in aspirates from the gastrointestinal tract. Only data obtained in this way provide a valid basis for setting the physiological parameters in physiologically based pharmacokinetic models.


Assuntos
Bicarbonatos/química , Ácidos e Sais Biliares/química , Líquidos Corporais/química , Líquidos Corporais/metabolismo , Trato Gastrointestinal Superior/química , Trato Gastrointestinal Superior/metabolismo , Tampões (Química) , Famotidina/administração & dosagem , Famotidina/metabolismo , Absorção Gastrointestinal , Humanos , Concentração de Íons de Hidrogênio , Ibuprofeno/administração & dosagem , Ibuprofeno/metabolismo , Intestino Delgado , Sais/química , Estômago
17.
Biosens Bioelectron ; 153: 112034, 2020 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-31989946

RESUMO

State of the art minimally invasive treatments and diagnostics of neurological and cardiovascular diseases demand for flexible instruments and implants that enable sensing and stimulation of bioelectric signals. Besides medical applications, implantable bioelectronic brain-computer interfaces are envisioned as the next step in communication and data transfer. Conventional microelectrode arrays used for these types of applications are based on polymer substrates that are not suitable for biostable, rigid and self-expanding devices. Here, we present fully integrated bioelectrodes on superelastic NiTi carriers fabricated by microsystem technology processes. The insulation between the metallic NiTi structure and the Pt electrode layer is realized by different oxide layers (SiOx, TaOx and Yttrium stabilized Zirconia YSZ). Key properties of bioelectronic implants such as dissolution in body fluids, biocompatibility, mechanical properties and bioelectrical sensing/stimulation capabilities have been investigated by in vitro methods. Particular devices with YSZ are biostable and biocompatible, enabling sensing and stimulation. The major advantage of this system is the combination of medically approved materials and novel fabrication technology that enables miniaturization and integration beyond the state-of-the-art processes. The results demonstrate that this functionalization of superelastic NiTi is an enabling technology for the development of new kinds of bioelectronic devices.


Assuntos
Técnicas Biossensoriais/instrumentação , Técnicas Eletroquímicas/métodos , Eletrodos Implantados , Microeletrodos , Níquel/química , Titânio/química , Ligas/química , Animais , Materiais Biocompatíveis/química , Líquidos Corporais/metabolismo , Equipamentos e Provisões , Humanos , Fenômenos Mecânicos , Microtecnologia , Óxidos/química , Polímeros/química , Próteses e Implantes , Propriedades de Superfície
18.
Biomed Pharmacother ; 123: 109801, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31901717

RESUMO

The aim of this study was to investigate the effect of Gotfried positive reduction (GPR) on repair of femoral neck fracture in rabbits and its underlying mechanisms. Male New Zealand white rabbits were employed to establish the model of femoral neck fracture. All the rabbits were randomly divided into four groups: control, open accurate reduction (OR), closed Gotfried negative support reduction (CR-N) and closed Gotfried positive support reduction (CR-P). At the 8th and 12th week after surgery, the anteroposterior and lateral radiographs of their hip joints were taken by X-ray, and local hemodynamics of their hip joints was detected by ultrasound. Histological examination was evaluated by HE staining and bone biological strength test was measured by testing machine, which was performed to study the repair of femoral neck fracture. Osteogenesis and angiogenesis-related proteins were measured by western blot in bone tissues and synovial tissues. The results revealed that the fracture healing intensity and blood supply in CR-P were better than those in CR-N and much more excellent than those in OR. In addition, the content of bone morphogenetic protein2 (BMP2), platelet derived growth factor (PDGF) and ocsteocalcin was higher in CR-P group than in CR-N, while lower in CR-P than OR. Furthermore, the expression of BMP2, COL-2 and angiopoietin (ANGPT) was upregulated in CR-P compared to CR-N and OR. Taken together, our results indicated that GPR was able to promote the repair of femoral neck fracture via enhancing osteogenesis and angiogenesis, which is valuable to us and shows good application prospect in bone tissue repair.


Assuntos
Fraturas do Colo Femoral/fisiopatologia , Fraturas do Colo Femoral/cirurgia , Consolidação da Fratura , Neovascularização Fisiológica , Osteogênese , Angiopoietinas/metabolismo , Animais , Fenômenos Biomecânicos , Líquidos Corporais/metabolismo , Proteínas Morfogenéticas Ósseas/metabolismo , Colágeno Tipo II/metabolismo , Fraturas do Colo Femoral/sangue , Fraturas do Colo Femoral/diagnóstico por imagem , Hemodinâmica , Osteocalcina/sangue , Fator de Crescimento Derivado de Plaquetas/metabolismo , Coelhos
19.
Nat Rev Cancer ; 20(2): 107-124, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31780785

RESUMO

Metastasis is a dynamic succession of events involving the dissemination of tumour cells to distant sites within the body, ultimately reducing the survival of patients with cancer. To colonize distant organs and, therefore, systemically disseminate within the organism, cancer cells and associated factors exploit several bodily fluid systems, which provide a natural transportation route. Indeed, the flow mechanics of the blood and lymphatic circulatory systems can be co-opted to improve the efficiency of cancer cell transit from the primary tumour, extravasation and metastatic seeding. Flow rates, vessel size and shear stress can all influence the survival of cancer cells in the circulation and control organotropic seeding patterns. Thus, in addition to using these fluids as a means to travel throughout the body, cancer cells exploit the underlying physical forces within these fluids to successfully seed distant metastases. In this Review, we describe how circulating tumour cells and tumour-associated factors leverage bodily fluids, their underlying forces and imposed stresses during metastasis. As the contribution of bodily fluids and their mechanics raises interesting questions about the biology of the metastatic cascade, an improved understanding of this process might provide a new avenue for targeting cancer cells in transit.


Assuntos
Líquidos Corporais/metabolismo , Modelos Biológicos , Neoplasias/metabolismo , Neoplasias/patologia , Microambiente Tumoral , Animais , Biomarcadores , Líquidos Corporais/efeitos dos fármacos , Matriz Extracelular/metabolismo , Humanos , Terapia de Alvo Molecular , Metástase Neoplásica , Neoplasias/etiologia , Neoplasias/terapia , Células Neoplásicas Circulantes/efeitos dos fármacos , Células Neoplásicas Circulantes/metabolismo , Células Neoplásicas Circulantes/patologia , Microambiente Tumoral/efeitos dos fármacos , Microambiente Tumoral/genética , Microambiente Tumoral/imunologia
20.
Nat Protoc ; 15(1): 40-67, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31776460

RESUMO

Gram-negative and Gram-positive bacteria release a variety of membrane vesicles through different formation routes. Knowledge of the structure, molecular cargo and function of bacterial extracellular vesicles (BEVs) is primarily obtained from bacteria cultured in laboratory conditions. BEVs in human body fluids have been less thoroughly investigated most probably due to the methodological challenges in separating BEVs from their matrix and host-derived eukaryotic extracellular vesicles (EEVs) such as exosomes and microvesicles. Here, we present a step-by-step procedure to separate and characterize BEVs from human body fluids. BEVs are separated through the orthogonal implementation of ultrafiltration, size-exclusion chromatography (SEC) and density-gradient centrifugation. Size separates BEVs from bacteria, flagella and cell debris in stool; and blood cells, high density lipoproteins (HDLs) and soluble proteins in blood. Density separates BEVs from fibers, protein aggregates and EEVs in stool; and low-density lipoproteins (LDLs), very-low-density lipoproteins (VLDLs), chylomicrons, protein aggregates and EEVs in blood. The procedure is label free, maintains the integrity of BEVs and ensures reproducibility through the use of automated liquid handlers. Post-separation BEVs are characterized using orthogonal biochemical endotoxin and Toll-like receptor-based reporter assays in combination with proteomics, electron microscopy and nanoparticle tracking analysis (NTA) to evaluate BEV quality, abundance, structure and molecular cargo. Separation and characterization of BEVs from body fluids can be done within 72 h, is compatible with EEV analysis and can be readily adopted by researchers experienced in basic molecular biology and extracellular vesicle analysis. We anticipate that this protocol will expand our knowledge on the biological heterogeneity, molecular cargo and function of BEVs in human body fluids and steer the development of laboratory research tools and clinical diagnostic kits.


Assuntos
Bactérias/citologia , Bioensaio/métodos , Líquidos Corporais/metabolismo , Vesículas Extracelulares/metabolismo , Humanos , Fatores de Tempo
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