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1.
Medicine (Baltimore) ; 100(11): e24852, 2021 Mar 19.
Artigo em Inglês | MEDLINE | ID: mdl-33725957

RESUMO

ABSTRACT: We evaluated the capacity of the XN-350 instrument to analyze 3 different types of body fluid samples under "body fluid mode."The performance of XN-350 was evaluated in terms of precision, carryover, limit of blank, limit of detection, limit of quantification, and linearity. Cell enumeration and differential data produced by the XN-350 were compared to manual chamber counting results in 63 cerebrospinal fluid (CSF), 51 ascitic fluid, and 51 pleural fluid (PF) samples. Comparisons between XN-350 versus Cytospin data were also performed in PF samples.The precision, carry-over, limit of blank, and linearity of the XN-350 were acceptable. The limits of detection for white blood cells (WBCs) and red blood cells were 1.0/µL, and 1,000.0/µL, respectively; the corresponding limits of quantitation (LOQs) were 5.0/µL and 2,000.0/µL, respectively. The XN-350's cell enumeration and differential counting correlated well with those of manual chamber counting for all 3 sample types (except for differential counting in CSF samples), particularly parameters involving monocytes (r = 0.33) and mononuclear cells (MO- body fluid [BF]; r = 0.26), as well as total cell (TC-BF) enumeration (r = 0.50) and WBC-BF (r = 0.50) in PF samples. The MO-BF in CSF samples differed significantly from manual chamber counting results, but neither TC-BF nor WBC-BF in PF samples did. The XN-350 also showed good correlations with Cytospin analyses for differential counting of neutrophils, lymphocytes, and monocytes in PF samples. The differential counting of eosinophils via the XN-350 and Cytospin were not significantly correlated, but the difference between them was not significant.The XN-350 is an acceptable alternative to manual fluid analysis. Samples with low cellularity around the LOQ should be checked manually. Moreover, manual differential counting should be performed on CSF samples, particularity those with low cell numbers.


Assuntos
Líquidos Corporais/química , Líquidos Corporais/citologia , Técnicas Citológicas/métodos , Testes Hematológicos/métodos , Microscopia/métodos , Líquido Ascítico/química , Líquido Ascítico/citologia , Líquido Cefalorraquidiano/química , Líquido Cefalorraquidiano/citologia , Humanos , Pleura/citologia , Pleura/metabolismo , Reprodutibilidade dos Testes
2.
Eur Rev Med Pharmacol Sci ; 24(23): 12516-12521, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-33336771

RESUMO

OBJECTIVE: The coronavirus disease 2019 (COVID-19) pandemic has significantly affected health care organizations globally. Many aspects of this disease, as well as the risks for patients treated with multiple drug regimens to control severe COVID-19, are unclear. During emergency surgery for SARS-CoV-2-positive patients, the risk of SARS-CoV-2 exposure and transmission to the surgical staff has yet to be determined. PATIENTS AND METHODS: In this report, we describe a SARS-CoV-2-positive patient with severe respiratory syndrome treated with multiple doses of IL-6 inhibitors who presented with a perforated duodenal ulcer and underwent emergency surgery. During and after surgery, we tested for SARS-CoV-2 at the ulcer site and in the peritoneal fluid. RESULTS: The history of the patient allows for two possible interpretations of the pathogenesis of the duodenal ulcer, which could have been a stress ulcer, or a gastrointestinal ulcer associated to the use of IL-6 inhibitors. We also noticed that the ulcer site and peritoneal fluid repeatedly tested negative for SARS-CoV-2. Therefore, we reviewed the pertinent literature on gastrointestinal bleeding in patients with COVID-19 and on SARS-CoV-2 detection in the peritoneal fluid of surgical patients and discussed possible prevention strategies for bleeding and the actual risk of infection for the surgical staff. CONCLUSIONS: The first implication of this case is that the relation between repeated administration of IL-6 inhibitors and upper gastrointestinal bleeding and perforation must be investigated, and that the threshold for administering prophylactic proton pump inhibitors therapy should be carefully considered for patients with severe COVID-19. The second implication is that further testing should be performed on the peritoneal fluid of COVID-19 patients undergoing emergency surgical procedures to clarify the discordant results for the presence of SARS-CoV-2 in the peritoneal cavity and the possible risk of transmission to the surgical staff.


Assuntos
Anticorpos Monoclonais Humanizados/efeitos adversos , Úlcera Duodenal/cirurgia , Úlcera Péptica Hemorrágica/cirurgia , Úlcera Péptica Perfurada/cirurgia , Estresse Fisiológico , Idoso , Líquido Ascítico/química , Líquido Ascítico/virologia , Úlcera Duodenal/virologia , Humanos , Masculino , Úlcera Péptica Hemorrágica/virologia , Úlcera Péptica Perfurada/virologia , RNA Viral/análise
3.
Khirurgiia (Mosk) ; (10): 36-43, 2020.
Artigo em Russo | MEDLINE | ID: mdl-33047584

RESUMO

OBJECTIVE: To determine the effect of intraperitoneal chemotherapy (IPC) with mitomycin C on expression of intraperitoneal cancer cells markers in patients with T4 colon cancer. MATERIAL AND METHODS: For the period from January 2019 to April 2020, 65 patients with T4 colon cancer were included in prospective comparative study. There were 46 patients in the main group and 19 patients in the control group. In the main group, surgical procedure was followed by IPC with mitomycin C. No IPC was performed in the control group. An effectiveness of IPC was evaluated using CD133, CD24, CD26, CD44, CD184 markers expression in peritoneal lavages. RESULTS: Significant between-group differences were observed for CD133 (p=0.0168), CD24 (p=0.0455) and CD44 (p=0.0012). There was a tendency to decrease in the level of CD184 expression in both groups in the second lavage (p=0.0605). CONCLUSION: IPC in patients with T4 colon cancer can reduce the expression and proliferative potential of free cancer cells.


Assuntos
Antibióticos Antineoplásicos/administração & dosagem , Neoplasias do Colo/tratamento farmacológico , Mitomicina/administração & dosagem , Antígeno AC133/análise , Antígeno AC133/biossíntese , Líquido Ascítico/química , Antígeno CD24/análise , Antígeno CD24/biossíntese , Proliferação de Células , Neoplasias do Colo/metabolismo , Neoplasias do Colo/patologia , Neoplasias do Colo/cirurgia , Dipeptidil Peptidase 4/análise , Dipeptidil Peptidase 4/biossíntese , Humanos , Receptores de Hialuronatos/análise , Receptores de Hialuronatos/biossíntese , Infusões Parenterais , Lavagem Peritoneal , Estudos Prospectivos , Receptores CXCR4/análise , Receptores CXCR4/biossíntese
4.
PLoS One ; 15(8): e0238119, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32845896

RESUMO

Abdominal tuberculosis (ATB) continues to pose a major diagnostic challenge for clinicians due to its nonspecific clinical presentation, variable anatomical location and lack of sensitive diagnostic tools. In spite of the development of several assays till date; no single test has proved to be adequate for ATB diagnosis. In this study, we for the first time report the detection of circulating cell-free Mycobacterium tuberculosis (M. tuberculosis) DNA (cfMTB-DNA) in ascitic fluid (AF) samples and its utility in ATB diagnosis. Sixty-five AF samples were included in the study and processed for liquid culture, cytological, biochemical and molecular assays. A composite reference standard (CRS) was formulated to categorize the patients into 'Definite ATB' (M. tuberculosis culture positive, n = 2), 'Probable ATB' (n = 16), 'Possible ATB' (n = 13) and 'Non-TB' category (n = 34). Two molecular assays were performed, namely, the novel cfMTB-DNA qPCR assay targeting M. tuberculosis devR gene and Xpert MTB/RIF assay (Xpert), and their diagnostic accuracy was assessed using CRS as reference standard. Clinical features such as fever, loss of weight, abdominal distension and positive Mantoux were found to be strongly associated with ATB disease (p<0.05). cfMTB-DNA qPCR had a sensitivity of 66.7% (95% CI:40.9,86.7) with 97.1% specificity (95% CI:84.7,99.9) in 'Definite ATB' and 'Probable ATB' group collectively. The sensitivity increased to 70.9% (95% CI:51.9,85.8) in the combined 'Definite', 'Probable' and 'Possible' ATB group with similar specificity. The cfMTB-DNA qPCR assay performed significantly better than the Xpert assay which demonstrated a poor sensitivity of ≤16.7% with 100% (95% CI:89.7,100) specificity (p<0.001). We conclude that cfMTB-DNA qPCR assay is an accurate molecular test that can provide direct evidence of M. tuberculosis etiology and has promise to pave the way for improving ATB diagnosis.


Assuntos
Líquido Ascítico/química , Ácidos Nucleicos Livres/análise , DNA Bacteriano/análise , Técnicas de Diagnóstico Molecular/métodos , Mycobacterium tuberculosis/genética , Tuberculose/diagnóstico , Abdome/microbiologia , Abdome/patologia , Adolescente , Adulto , Idoso , Proteínas de Bactérias/genética , Proteínas de Ligação a DNA/genética , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase em Tempo Real , Tuberculose/patologia , Adulto Jovem
5.
PLoS One ; 15(5): e0233350, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32437441

RESUMO

BACKGROUND: Serum-ascites albumin gradient (SAAG) remains the most sensitive and specific marker for the differentiation of ascites due to portal hypertension from ascites due to other causes. SAAG has some limitations and may fail in selected conditions. Voltammetric analysis (VA) has been used for the detection of electroactive species of biological significance and has proven effective for detection infections in biological fluids. AIMS: In this study, we compared the accuracy of voltammetric analysis (VA) with that of SAAG to differentiate ascites due to portal hypertension from that having a different origin. METHODS: 80 ascites samples were obtained from patients undergoing paracentesis at the Campus Bio-Medico Hospital of Rome. VA was performed using the BIONOTE device. The ability of VA to discriminate ascitic fluid etiology and biochemical parameters was evaluated using Partial Least Square Discriminant Analysis (PLS-DA), with ten-fold cross-validations. RESULTS: Mean age was 68.6 years (SD 12.5), 58% were male. Ascites was secondary to only portal hypertension in 72.5% of cases (58 subjects) and it was secondary to a baseline neoplastic disease in 27.5% of cases (22 subjects). Compared to SAAG≥1.1, e-tongue predicted ascites from portal hypertension with a better accuracy (92.5% Vs 87.5%); sensitivity (98.3% Vs 94.8%); specificity (77.3% Vs 68.2%); predictive values (PPV 91.9% Vs 88.7% and NPV 94.4% Vs 83.3%). VA correctly classified ascites etiology in 57/58 (98.2%) of cases with portal hypertension and in 17/22 (77.2%) of cases with malignancy. Instead, VA showed poor predictive capacities towards total white blood count and polymorphonuclear cell count. CONCLUSIONS: According to this proof of concept study, VA qualifies as a promising low-cost and easy method to discriminate between ascites secondary to portal hypertension and ascites due to malignancy.


Assuntos
Ascite/diagnóstico , Ascite/etiologia , Técnicas Eletroquímicas/métodos , Hipertensão Portal/complicações , Hipertensão Portal/diagnóstico , Neoplasias/complicações , Neoplasias/diagnóstico , Idoso , Idoso de 80 Anos ou mais , Albuminas/análise , Líquido Ascítico/química , Biomarcadores/análise , Técnicas de Química Analítica/métodos , Técnicas de Química Analítica/estatística & dados numéricos , Diagnóstico Diferencial , Técnicas Eletroquímicas/estatística & dados numéricos , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Reconhecimento Automatizado de Padrão , Estudo de Prova de Conceito , Albumina Sérica Humana/análise
6.
Am Surg ; 86(4): 334-340, 2020 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-32391757

RESUMO

Different kinds of complications after splenectomy in hepatolenticular degeneration patients with hypersplenism have been reported in the past decades, but studies on pancreatic fistula and the corresponding targeted prevention and treatment after splenectomy still remain much unexplored. The present work investigated the pathogenic factors of pancreatic fistula after splenectomy and the variation tendency of amylase in drainage fluid, aiming to verify the significance of monitoring amylase in the abdominal drainage fluid in the early diagnosis of pancreatic fistula after splenectomy. One hundred sixty-seven patients with hepatolenticular degeneration and hypersplenism who underwent splenectomy in the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine from January 2016 to August 2018 were selected and analyzed. The amylase in the abdominal drainage fluid was monitored routinely after splenectomy. We also conducted the statistics on the incidence of different types of pancreatic fistula and analyzed the influence factors of pancreatic fistula formation. After splenectomy, biochemical fistula occurred in 11 patients (6.6%), grade B fistula in six patients (3.6%), grade C fistula in one patient (0.6%), and the incidence of pancreatic fistula was 4.2 per cent (biochemical fistula excluded). The amylase in the peritoneal drainage fluid was closely concerned with the incidence of pancreatic fistula according to our statistics. Furthermore, by analyzing the different influence factors of pancreatic fistula, Child-Pugh grading of liver function (P = 0.041), pancreatic texture (P = 0.029), degree of splenomegaly (P = 0.003), and operative method (P = 0.001) were supposed to be closely related to the formation of pancreatic fistula. Monitoring of amylase in peritoneal drainage fluid is regarded as an important physiological parameter in the early diagnosis of pancreatic fistula after splenectomy, which provides effective clinical reference and plays a significant role in preventing the occurrence and development of pancreatic fistula.


Assuntos
Amilases/análise , Líquido Ascítico/química , Degeneração Hepatolenticular/cirurgia , Fístula Pancreática/etiologia , Complicações Pós-Operatórias/epidemiologia , Esplenectomia/efeitos adversos , Esplenomegalia/cirurgia , Adolescente , Adulto , Idoso , Biomarcadores/análise , Criança , Drenagem , Feminino , Degeneração Hepatolenticular/complicações , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Monitorização Fisiológica , Fístula Pancreática/epidemiologia , Fístula Pancreática/prevenção & controle , Esplenomegalia/etiologia , Adulto Jovem
7.
Am J Clin Pathol ; 154(1): 103-114, 2020 06 08.
Artigo em Inglês | MEDLINE | ID: mdl-32271370

RESUMO

OBJECTIVES: High-grade serous carcinoma (HGSC) is the most common ovarian malignancy. The role of cytopathology in obtaining tissue diagnosis before institution of neoadjuvant chemotherapy (NACT) was evaluated. METHODS: All histopathology-proven HGSC specimens between 2015 and 2018 with prior cytopathologic diagnosis by ascitic fluid evaluation or fine-needle aspiration (FNA) of ovarian mass were reviewed with cell block immunocytochemistry for CK7, CK20, PAX8, WT1, and p53. RESULTS: Of 288 cases of HGSC, pre-NACT cytology diagnosis was established in 32% (93/288), with specific HGSC diagnoses made on ascitic fluid in 88% (82/93) and by ovarian mass FNA in 12% (11/93). The ascitic fluid showed moderate/high cellularity with papillary clusters in 76% (71/93) cases. Cell block immunocytochemistry showed tumor cells positive for CK7, PAX8, and WT1. p53 showed mutant or null-type positivity in 65% (33/51) and 33% (17/51) of cases, respectively, with 100% concordance with subsequent histopathology specimens. Poor/intermediate response to chemotherapy was shown in 75% of cases. CONCLUSIONS: Combined assessment of cytomorphology, cell block histomorphology, and ancillary immunohistochemical testing, including PAX8, WT1, and p53, allows for specific pre-NACT diagnoses of HGSC in ascitic fluid and ovarian FNA cytology. This practice allows for initiation of chemotherapy and diminution of disease burden prior to definitive surgical therapy.


Assuntos
Líquido Ascítico/patologia , Biomarcadores Tumorais/análise , Carcinoma Epitelial do Ovário/diagnóstico , Cistadenocarcinoma Seroso/diagnóstico , Citodiagnóstico/métodos , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Líquido Ascítico/química , Biópsia por Agulha Fina/métodos , Feminino , Humanos , Imuno-Histoquímica , Pessoa de Meia-Idade , Estudos Retrospectivos , Adulto Jovem
8.
In Vivo ; 34(2): 715-722, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32111775

RESUMO

BACKGROUND/AIM: There are two strategies for the interpretation of tumor markers (TM) in fluid effusions: i) high cut-off and ii) fluid/serum ratio (F/S) and low cut-off. The objective of this study is to compare these two strategies and to determine whether diagnostic accuracy improves by the identification of possible false positives using Adenosine deaminase (ADA), C reactive protein (CRP) and % of polymorphonuclear cells (%PN). PATIENTS AND METHODS: We studied 157 ascitic fluids, 74 of which were malignant. ADA, CRP and %PN were determined in ascitic fluid, and Carcinoembryonic antigen (CEA), Cancer antigen 72-4 (CA72-4), Cancer antigen CA19-9 and Cancer antigen 15-3 (CA15-3) in both fluid and serum. RESULTS: The strategy of high cut-off showed 59.5% sensitivity at 100% specificity. The F/S strategy showed 75.7% sensitivity at 95.2% specificity. Subclassifying cases with ADA, CRP and %PN negative showed 67.5% sensitivity at 100% specificity for high cut-off and for the F/S strategy was 81.7% sensitivity at 98.7% specificity. CONCLUSION: The strategy of F/S with negative ADA, CRP and %PN allow the best interpretation for TM in the ascitic fluid.


Assuntos
Líquido Ascítico/metabolismo , Biomarcadores Tumorais/sangue , Neoplasias/sangue , Adenosina Desaminase/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Antígenos Glicosídicos Associados a Tumores/metabolismo , Líquido Ascítico/química , Biomarcadores Tumorais/análise , Proteína C-Reativa/metabolismo , Antígeno CA-19-9/metabolismo , Antígeno Carcinoembrionário/metabolismo , Diagnóstico Diferencial , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Mucina-1/metabolismo , Neoplasias/diagnóstico , Neoplasias/metabolismo , Neutrófilos/patologia , Sensibilidade e Especificidade
9.
BMC Cancer ; 20(1): 225, 2020 Mar 17.
Artigo em Inglês | MEDLINE | ID: mdl-32178642

RESUMO

BACKGROUND: Considering the potential of p16 as a marker for diagnosis, prognosis and therapeutic response, the aim of this study was to assess its presence, via immunocytochemistry, in metastatic carcinoma of different primary sites and histological types obtained from effusions and peritoneal washings. A total of 118 samples including 85 of metastatic carcinoma and 33 samples of benign effusion/peritoneal washing were prepared by the plasma/thromboplastin method. Immunocytochemistry reactions were performed on cell block sections using antibodies against p16, claudin-4, MOC-31, calretinin, HBME and CD68. RESULTS: P16 overexpression was observed in 88.23% of all carcinoma samples. All cervix adenocarcinoma samples showed p16 overexpression. Overexpression in adenocarcinomas of ovary, lung and breast was observed in 93.75, 93.10 and 75% of the samples, respectively. Overexpression was observed in all different histological types analyzed: small cell carcinoma (lung), squamous cell carcinoma (cervical) and urothelial carcinoma (bladder). The specificity of p16 for carcinoma detection was of 96.96%. CONCLUSION: Overexpression of p16 was observed in most metastatic carcinoma, from different primary sites and histological types, obtained from effusions and peritoneal washings. Due to its high frequency of overexpression in metastatic carcinoma, p16 may play a possible role in tumor progression and it may be considered as a complementary diagnostic marker depending on histological type and primary site of carcinoma.


Assuntos
Líquido Ascítico/química , Biomarcadores Tumorais/análise , Carcinoma/diagnóstico , Carcinoma/secundário , Inibidor p16 de Quinase Dependente de Ciclina/análise , Neoplasias/diagnóstico , Neoplasias/patologia , Derrame Pericárdico/química , Derrame Pleural Maligno/química , Antígenos CD/análise , Antígenos CD/imunologia , Antígenos de Diferenciação Mielomonocítica/análise , Antígenos de Diferenciação Mielomonocítica/imunologia , Antígenos de Superfície/análise , Antígenos de Superfície/imunologia , Biomarcadores Tumorais/imunologia , Calbindina 2/análise , Calbindina 2/imunologia , Claudina-4/análise , Claudina-4/imunologia , Inibidor p16 de Quinase Dependente de Ciclina/imunologia , Humanos , Prognóstico
10.
Pesqui. vet. bras ; 40(3): 158-164, Mar. 2020. tab
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1135602

RESUMO

This study aimed to evaluate the appropriate sites of abdominocentesis for peritoneal fluid collection in cattle and to investigate the time of cell viability in vitro, comparing three methods of sample conservation. Twenty-one healthy cattle (19 females and 2 males) were subjected to a laparocentesis procedure to obtain peritoneal fluid, with punctures in three defined sites: left cranial, right cranial, and right caudal. The total peritoneal fluid collected was divided into three aliquots and maintained under three preservation conditions: room temperature (26°C), refrigeration (4°C), and room temperature (26°C) with the addition of 1µL of 10% formaldehyde per 1mL of peritoneal fluid. The peritoneal fluid analysis performed immediately after collection consisted of: physical examination (color, appearance, volume, and specific gravity), biochemical measures (pH, total protein, fibrinogen, creatinine, and glucose), and cellularity (total and differential counts). The determination of proteins and the examination of cells were repeated in each separate aliquot at two, four, six, and eight hours after harvest. Data were analyzed through repeated measures ANOVA or Friedman test. The harvest was productive in 67% of cattle. The left cranial and the right cranial puncture sites were the most appropriate. Peritoneal fluid analyzed after collection, the total protein concentration ranged from 1.4 to 3.6g/dL, and number of leukocytes ranged from 54 to 1,322 cells/µL; 60 to 95% of leukocytes were lymphocytes. The protein concentration decreased, but the absolute values of leukocytes, lymphocytes, and segmented neutrophils did not change up to eight hours after collection, independent of the maintenance method. Cell lysis was delayed by cooling, and the addition of formaldehyde did not help preserve the integrity of cellular morphology. Laparocentesis is a safe and secure procedure in cattle and maybe more productive when performed in specific sites on the left or right sides of the cranial abdominal wall. Peritoneal fluid samples may be analyzed with reliable results for up to eight hours after collection when kept refrigerated and for up to six hours when kept at room temperature.(AU)


O estudo teve como objetivo avaliar os locais adequados de laparocentese para a colheita de fluido peritoneal de bovinos e estabelecer o tempo de viabilidade celular in vitro, comparando três métodos de conservação. Vinte e um bovinos hígidos (19 fêmeas e 2 machos) foram submetidos ao procedimento de laparocentese para obtenção de fluido peritoneal, com punção em três pontos definidos: cranial esquerdo, cranial direito e caudal direito. O volume total do líquido peritoneal foi dividido em três alíquotas mantidas sob três métodos de conservação: temperatura ambiente (26°C); refrigeração (4°C); e temperatura ambiente (26°C) com adição de 1µL de formol 10% para cada 1mL de líquido peritonial. A análise do líquido peritoneal realizada imediatamente após sua obtenção consistiu em: exames físico (cor, aspecto, volume e densidade); bioquímicos (pH, proteína total, fibrinogênio, creatinina e glicose); e da celularidade (contagens total e diferencial). A determinação de proteínas e o exame da celularidade foram repetidos, em cada alíquota separada, as duas, quatro, seis e oito horas após a colheita. Análise de variâncias de medidas repetidas ou teste de Friedman foram empregados para avaliação ao longo do tempo. A colheita foi produtiva em 67% dos bovinos e os locais de punção craniais esquerdo e direito foram os mais adequados. A concentração de proteína total variou de 1,4 a 3,6g/dL e o número de leucócitos de 54 a 1.322 células/µL, com predomínio de linfócitos (60 a 95% das células) no fluido peritoneal analisado logo após a colheita. A concentração de proteínas diminuiu, mas os valores absolutos de leucócitos, de linfócitos e de neutrófilos segmentados não se modificaram até oito horas após a colheita, independente do método de manutenção das amostras. A lise celular foi retardada pela refrigeração e a adição de formol não contribuiu para preservar a integridade da morfologia celular. A laparocentese é um procedimento seguro e de execução fácil em bovinos sendo mais produtiva quando realizada em locais específicos à esquerda ou à direita craniais da parede abdominal. Amostras de fluido peritoneal podem ser analisadas com resultados confiáveis quando mantidas refrigeradas por até oito horas após a colheita e quando mantidas à temperatura ambiente por até seis horas.(AU)


Assuntos
Animais , Bovinos , Líquido Ascítico/citologia , Líquido Ascítico/química , Punções/métodos , Cavidade Abdominal/patologia , Peritonite/diagnóstico
11.
Fertil Steril ; 113(2): 364-373.e2, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-32106990

RESUMO

OBJECTIVE: To demonstrate the feasibility of studying exosomes directly from peritoneal fluid, we isolated exosomes from endometriosis patient samples and from controls, and characterized their cargo. DESIGN: Case-control experimental study. SETTING: Academic clinical center. PATIENT (S): Women with and without endometriosis who underwent laparoscopic surgery (n = 28 in total). INTERVENTION (S): None. MAIN OUTCOME MEASURE (S): Concentration of exosomes within peritoneal fluid and protein content of the isolated exosomes. RESULT (S): Peritoneal fluid samples were pooled according to the cycle phase and disease stage to form six experimental groups, from which the exosomes were isolated. Exosomes were successfully isolated from peritoneal fluid in all the study groups. The concentration varied with cycle phase and disease stage. Proteomic analysis showed specific proteins in the exosomes derived from endometriosis patients that were absent in the controls. Five proteins were found exclusively in the endometriosis groups: PRDX1, H2A type 2-C, ANXA2, ITIH4, and the tubulin α-chain. CONCLUSION (S): Exosomes are present in peritoneal fluid. The characterization of endometriosis-specific exosomes opens up new avenues for the diagnosis and investigation of endometriosis.


Assuntos
Líquido Ascítico/química , Endometriose/metabolismo , Exossomos/química , Proteínas/análise , Adulto , Anexina A2/análise , Líquido Ascítico/patologia , Estudos de Casos e Controles , Endometriose/patologia , Exossomos/ultraestrutura , Estudos de Viabilidade , Feminino , Histonas/análise , Humanos , Pessoa de Meia-Idade , Peroxirredoxinas/análise , Proteínas Secretadas Inibidoras de Proteinases/análise , Proteômica , Tubulina (Proteína)/análise , Adulto Jovem
12.
Ann Biol Clin (Paris) ; 78(1): 17-26, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-32108576

RESUMO

The lipoproteinogram (or lipidogram) consists in an electrophoretic separation of the main classes of serum lipoproteins. Separation was done in agarose gel using the Sebia Hydragel Lipo + Lp(a)® kit. A repeatability study (n=6) was conducted on 3 sera (1 normolipidemic, 1 hypertriglyceridemic and 1 with a high Lp(a) concentration). The reproducibility was studied on these 3 sera and on an ascites liquid containing chylomicrons, upon 6 days (n=6). A quantitative approach was made by studying areas under the curve and percentages of fractions. In both cases (repeatability and reproducibility), the revelation of the lipoproteins in the gel after electrophoretic migration was made either by staining with Sudan Black (procedure recommended by Sebia), or with Fat Red 7B. Regardless of staining, both repeatability and reproducibility studies show that all lipoprotein fractions were correctly detected at their respective positions, leading to satisfactory interpretations of lipoproteinograms. Our reproducibility study also confirmed a good stability of the fractions over 6 days (storage at +5 ± 3̊C). In addition, the Fat Red 7B staining leads to a shorter technical time (about 40 min) for the gel drying and staining/destaining phases, which allows us to respond more quickly to certain urgent requests such as chylothorax diagnosis.


Assuntos
Compostos Azo/farmacologia , Eletroforese/métodos , Lipoproteínas/análise , Lipoproteínas/sangue , Kit de Reagentes para Diagnóstico , Coloração e Rotulagem/métodos , Líquido Ascítico/química , Compostos Azo/química , Análise Química do Sangue/métodos , Fracionamento Químico/métodos , Eletroforese em Gel de Ágar , Humanos , Lipoproteína(a)/análise , Lipoproteína(a)/sangue , Reprodutibilidade dos Testes
14.
Artigo em Inglês | MEDLINE | ID: mdl-31958565

RESUMO

The echinocandins anidulafungin (ANID) and micafungin (MICA) are recommended for treatment of invasive Candida infections. As target-site concentrations of antimicrobial agents are crucial for eradication of pathogens, we established and validated high-performance liquid chromatography-UV detection (HPLC-UV) assays for quantification of ANID and MICA in human plasma, ascites fluid, pleural effusion, and in cerebrospinal fluid (CSF). Sample pre-purification was performed by protein precipitation with acetonitrile followed by solid phase extraction. For both assays, intra- and interday precision, and accuracy fulfilled the requirements for bioanalytical methods issued by the European Medicine Agency (EMA). The lower limit of quantification was 0.01 mg/L for both drugs. At 25 °C, ANID and MICA concentrations declined by up to 70% within 24 h. Concentrations remained stable over 24 h at 4 °C and over four weeks at -80 °C. In conclusion, the developed methods are fit for the assessment of target-site pharmacokinetics of ANID and MICA in clinical studies.


Assuntos
Anidulafungina/análise , Cromatografia Líquida de Alta Pressão/métodos , Micafungina/análise , Espectrofotometria Ultravioleta/métodos , Anidulafungina/uso terapêutico , Líquido Ascítico/química , Candidíase Invasiva/tratamento farmacológico , Monitoramento de Medicamentos/métodos , Humanos , Limite de Detecção , Modelos Lineares , Micafungina/uso terapêutico , Reprodutibilidade dos Testes , Extração em Fase Sólida
15.
Acta Cytol ; 64(3): 248-255, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31352449

RESUMO

BACKGROUND: Differentiating reactive mesothelial cells from metastatic carcinoma in effusion cytology is a challenging task. The application of at least 4 monoclonal antibodies including 2 epithelial markers (Ber-EP4, MOC-31, CEA, or B72.3) and 2 mesothelial markers (calretinin, WT-1, CK5/6, or HBME-1) are often useful in this distinction; however, it is not readily available in many resource-limited developing countries. Aberrant immunoexpression of enhancer of zeste homolog 2 (EZH2), a transcriptional repressor involved in cancer progression, is observed widely in various malignancy. In this study, we evaluate the diagnostic value of EZH2 as a single reliable immunomarker for malignancy in effusion samples. METHODS: A total of 108 pleural, peritoneal, and pericardial effusions/washings diagnosed as unequivocally reactive (n = 41) and metastatic carcinoma (n = 67) by cytomorphology over 18 months were reviewed. Among the metastatic carcinoma cases, 54 were adenocarcinoma and others were squamous cell carcinoma (n = 1), carcinosarcoma (n = 1), and carcinoma of undefined histological subtypes (n = 11). Cell block sections were immunostained by EZH2 (Cell Marque, USA). The percentages of EZH2-immunolabeled cells over the total cells of interest were calculated. Receiver operating characteristic (ROC) curve analysis was performed to determine the optimal cut-off score to define EZH2 immunopositivity. RESULTS: A threshold of 8% EZH2-immunolabeled cells allows distinction between malignant and reactive mesothelial cells, with 95.5% sensitivity, 100% specificity, 100% positive predictive value, and 93.2% negative predictive value (p < 0.0001). The area under the curve was 0.988. CONCLUSION: EZH2 is a promising diagnostic biomarker for malignancy in effusion cytology which is inexpensive yet trustworthy and could potentially be used routinely in countries under considerable economic constraints.


Assuntos
Líquido Ascítico/patologia , Biomarcadores Tumorais/análise , Carcinoma/diagnóstico , Proteína Potenciadora do Homólogo 2 de Zeste/análise , Derrame Pericárdico/diagnóstico , Derrame Pleural Maligno/diagnóstico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Líquido Ascítico/química , Carcinoma/complicações , Citodiagnóstico/métodos , Feminino , Humanos , Imuno-Histoquímica/métodos , Masculino , Pessoa de Meia-Idade , Derrame Pericárdico/etiologia , Derrame Pleural Maligno/etiologia , Estudos Retrospectivos , Adulto Jovem
16.
J Gastroenterol Hepatol ; 35(2): 271-277, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31247673

RESUMO

BACKGROUND AND AIMS: Diagnostic performance of ascitic fluid total protein (AFTP) concentration remained unsettled. Our aim was to determine diagnostic value of AFTP in differential diagnosis of causes of ascites. METHODS: Seven hundred four consecutive patients with new-onset ascites were prospectively enrolled in this study. RESULTS: In the training cohort, diagnostic performance of quantitative AFTP assay was superior to that of Rivalta test in differential diagnosis of ascites. At the predetermined cut-off value of 25 g/L, quantitative AFTP assay was more useful in the differentiation of non-portal hypertensive ascites from portal hypertensive ascites compared with the exudate-transudate classification, area under curve of receiver operating characteristic curve was 0.958. Quantitative AFTP assay was superior to serum-ascites albumin gradient in the detection of non-portal hypertensive ascites, especially malignant ascites and tuberculous peritonitis. In mixed ascites, AFTP was useful in identifying peritoneal lesions. CONCLUSIONS: Ascitic fluid total protein is a useful marker in non-portal hypertensive ascites; thus, it should be determined in diagnostic work-up of the patients with ascites.


Assuntos
Ascite/diagnóstico , Ascite/etiologia , Líquido Ascítico/química , Proteínas/análise , Biomarcadores/análise , Diagnóstico Diferencial , Humanos , Hipertensão/complicações
17.
Sci Rep ; 9(1): 17558, 2019 11 26.
Artigo em Inglês | MEDLINE | ID: mdl-31772282

RESUMO

Postoperative adhesion formation often ruins the quality of life or is an obstacle to illnesses with curative operation such as cancer. Previously we demonstrated that interferon-γ-promoted fibrin deposition drove postoperative adhesion formation. However, its underlying cellular and molecular mechanisms remain poorly understood. We found that myofibroblasts of the adhesion predominantly expressed signature molecules of mesothelial cells that line the serosa. Microarray analysis revealed IL-6 as a key underlying player, supported by elevated IL-6 levels in the peritoneal fluid of post-laparotomy human subjects. Injured serosa of cecum-cauterized mice also exhibited induction of Il6, which was followed by Tnf, concomitant with rapid accumulation of neutrophils, substantial population of which expressed TGF-ß1, a master regulator of fibrosis. Besides, neutrophil-ablated mice showed reduction in induction of the adhesion, suggesting that TGF-ß1+neutrophils triggered the adhesion. Human neutrophils expressed TGFB1 in response to TNF-α and TNF in response to IL-6. Moreover, anti-IL-6 receptor monoclonal antibody abrogated neutrophil recruitment and adhesion formation. Thus, IL-6 signaling represents a potential target for the prevention of postoperative adhesions.


Assuntos
Anticorpos Monoclonais/uso terapêutico , Complicações Pós-Operatórias/prevenção & controle , Receptores de Interleucina-6/imunologia , Aderências Teciduais/prevenção & controle , Animais , Anticorpos Monoclonais/imunologia , Líquido Ascítico/química , Humanos , Interleucina-6/análise , Interleucina-6/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Miofibroblastos/metabolismo , Neutrófilos/metabolismo , Receptores de Interleucina-6/fisiologia , Fator de Crescimento Transformador beta1/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
18.
Cancer Invest ; 37(9): 440-452, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31530033

RESUMO

Ovarian cancer is the deadliest gynecologic cancer. The large-scale microRNA (miRNA) expression profiling and individual miRNA validation was performed to find potential novel biomarkers for ovarian cancer. The most consistent overexpression of miRs-200b-3p, 135 b-5p and 182-5p was found in both ascitic fluid and tumors and suggests their potential as oncogenes. miR-451a was consistently underexpressed so may be a tumor suppressor. Results were inconsistent for miR-204-5p, which was overexpressed in ascitic fluid but underexpressed in tumor tissue. miR-203a-3p was generally overexpressed but this failed to be proved in independent sample set in tissue validation.


Assuntos
Líquido Ascítico/química , Biomarcadores Tumorais/genética , Perfilação da Expressão Gênica/métodos , MicroRNAs/genética , Neoplasias Ovarianas/genética , Ovário/química , Idoso , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Neoplasias Ovarianas/patologia , Prognóstico
19.
Ethiop J Health Sci ; 29(3): 383-390, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31447507

RESUMO

Background: The aim of this study was to assess the role of transabdominal ultrasonography in characterizing and determining the etiology of ascites in comparison with laboratory ascitic fluid analysis and other methods used to establish the final diagnosis. Methods: A prospective descriptive study was conducted on 61 patients with ascites attending outpatient department (OPD) or admitted to wards of Tikur Anbesa Specialized Hospital (TASH) and referred to radiology department for imaging from June 2017 to November 2017. Data were collected following the internationally recommended scanning technique in consecutive bases. The data were analyzed using SPSS version 20. The comparison of ultrasound and laboratory findings with final clinical diagnosis was analyzed using Chi-square test (X2). Results: Of 61 patients with ascites enrolled in this study, females were 35(57.4%) with age range of 16 to 75 and mean age of 43.2±14.11. The cause of ascites was established in 59 cases using a combination of clinical, pathological, imaging evidences and tumor markers. However there were two cases who had ascites with indeterminate cause. US suggested the diagnosis in 54(91.5%) patients. Excluding mixed and indeterminate cases, ultrasound characterized ascites correctly as exudate and transudate in 95% cases. Conclusion: Ultrasound has significant accuracy to distinguish transudate and exudate ascites and in suggesting the underlying cause. It can be a valuable method of investigation of ascites in places where CT and MRI are not available, and it is the best complement for laboratory investigations on ascites in suggesting the etiology based on ascitic fluid texture and ancillary findings.


Assuntos
Ascite/diagnóstico por imagem , Ultrassonografia , Adolescente , Adulto , Idoso , Ascite/diagnóstico , Ascite/etiologia , Líquido Ascítico/química , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Adulto Jovem
20.
Reprod Biomed Online ; 39(4): 704-711, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31427176

RESUMO

RESEARCH QUESTION: Immunological disorders have been reported to promote the progression of endometriosis. Several recent studies have shown that myeloid-derived suppressor cells (MDSC) drive the progression of endometriosis. The aim of this case-control study was to test whether CCR5 and its ligands drive MDSC accumulation and play a role in the progression of endometriosis. DESIGN: Thirty-six endometriosis patients and 20 controls were recruited. All subjects underwent laparoscopy. An ELISA kit was used to define CCR5 ligands in plasma and peritoneal fluid from endometriosis patients; flow cytometry was then used to characterize CCR5+MDSC in peripheral blood and peritoneal fluid. RESULTS: Data showed that endometriosis patients displayed a significantly higher production of plasma CCL3 (P = 0.046) and peritoneal fluid CCL3/5 (P = 0.042/0.036) compared with those from the uterine leiomyoma group. Furthermore, the concentrations of peritoneal fluid CCL5 were elevated in late stage patients compared with those from the uterine leiomyoma group. Accumulation of blood CCR5+Mo-MDSC was detected in endometriosis patients compared with those from both the ovarian dermoid cysts and uterine leiomyoma groups. Endometriosis patients also showed an elevation of CCR5+MDSC and CCR5+Mo-MDSC in peritoneal fluid samples compared with uterine leiomyoma samples. It was also found that enrichment of CCR5+MDSC (r = 0.6807; P < 0.0001) and CCR5+Mo-MDSC (r = 0.6893; P < 0.0001) were correlated with enhanced production of CCL5 in peritoneal fluid from endometriosis patients. CONCLUSIONS: This study showed that CCR5 and its ligands could drive the progression of endometriosis by enhancing the accumulation of MDSC. These findings might produce a promising treatment that targets CCR5+MDSC for endometriosis patients.


Assuntos
Quimiocina CCL4/metabolismo , Endometriose/patologia , Células Supressoras Mieloides/metabolismo , Doenças Peritoneais/patologia , Receptores CCR5/metabolismo , Adulto , Líquido Ascítico/química , Líquido Ascítico/metabolismo , Estudos de Casos e Controles , Quimiocina CCL3/sangue , Quimiocina CCL3/metabolismo , Quimiocina CCL4/sangue , Quimiocina CCL5/sangue , Quimiocina CCL5/metabolismo , Progressão da Doença , Endometriose/sangue , Endometriose/metabolismo , Feminino , Humanos , Ligantes , Células Supressoras Mieloides/fisiologia , Doenças Peritoneais/sangue , Doenças Peritoneais/metabolismo
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