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Fertility and Sterility Oct 2023To propose a standardized protocol for peritoneal free fluid and leukocyte sample collection in women with endometriosis suitable for biomedical research on the basis of...
OBJECTIVE
To propose a standardized protocol for peritoneal free fluid and leukocyte sample collection in women with endometriosis suitable for biomedical research on the basis of the surgical procedure, the clinical and technical conditions, and the quality of the samples obtained.
DESIGN
Video showing the step-by-step collection procedure and the suitability of samples obtained for biomedical research.
SUBJECTS
This study included 103 women with confirmed endometriosis by pathology analysis, who signed informed consent and were recruited from the Hospital "Virgen de la Arrixaca", Murcia, Spain. The study was approved by the Ethics Committee of University of Murcia (CEI 3156/2020).
MAIN OUTCOME MEASURES
We analyzed the presence of free fluid in the peritoneal cavity and its relationship with hormonal treatment intake. In addition, the presence of blood contamination, the number of viable leukocytes and macrophages in free peritoneal fluid and lavages as well as their relationship with the lavage volume used, the body mass index, and the age of patients were analyzed.
RESULTS
The presence of free peritoneal fluid, in which cells and molecules could be quantified, was scarce in the patients (21%), and it was not significantly related to hormonal treatment intake. The cell viability was higher than 98% in all collected samples; although 54% showed good quality and enough cellularity to be used in biomedical research, 40% were contaminated with blood and 6% had low cellularity. The number of leukocytes and macrophages recovered from the peritoneal lavages correlated positively with the lavage volume used and negatively with the body mass index and was independent of the age of the patients.
CONCLUSION
We describe a standardized step-by-step procedure for peritoneal fluid and leukocyte collection in women with endometriosis, suitable for biomedical research, taking into account that not all women present free fluid in the peritoneal cavity. We propose to increase the lavage volume recommended by the World Endometriosis Research Foundation from 10 mL to at least 40 mL of sterile saline solution and its mobilization for at least 30 seconds within the peritoneal cavity, especially in patients with higher body mass index, to improve the efficiency of the procedure.
Topics: Humans; Female; Endometriosis; Ascitic Fluid; Leukocytes; Peritoneum; Macrophages
PubMed: 37392781
DOI: 10.1016/j.fertnstert.2023.06.030 -
Methods in Molecular Biology (Clifton,... 2019There is limited information regarding the biobanking of pleural and peritoneal fluids that might supplement storage of pulmonary and thoracic tissue biospecimens. Such... (Review)
Review
There is limited information regarding the biobanking of pleural and peritoneal fluids that might supplement storage of pulmonary and thoracic tissue biospecimens. Such fluids are sometimes collected for clinical analyses and may have uses that obviate or supplement tissue samples. There has been a growing interest in using liquid biopsies as they are less invasive and may be amenable to analyses that guide targeted therapies. Integrating cytology and biobanking approaches, we describe techniques that may be used for collecting and banking pleural and peritoneal fluids.
Topics: Ascitic Fluid; Biological Specimen Banks; Humans; Specimen Handling; Tissue Banks
PubMed: 30539441
DOI: 10.1007/978-1-4939-8935-5_13 -
Reproductive Biomedicine Online Oct 2021What are the potential biomarkers for peritoneal endometriosis in peritoneal fluid and serum?
RESEARCH QUESTION
What are the potential biomarkers for peritoneal endometriosis in peritoneal fluid and serum?
DESIGN
Case-control studies composed of independent discovery and validation sets were conducted. In the discovery set, untargeted liquid chromatography-mass spectrometry (LC-MS/MS) metabolomics, multivariable and univariable analyses were conducted to generate global metabolomic profiles of peritoneal fluid for endometriosis and to identify potential metabolites that could distinguish peritoneal endometriosis (n = 10) from controls (n = 31). The identified metabolites from the discovery set were validated in independent peritoneal fluid (n =19 peritoneal endometriosis and n = 20 controls) and serum samples (n = 16 peritoneal endometriosis and n = 19 controls) using targeted metabolomics. The area under the receiver-operating characteristics curve (AUC) analysis was used to evaluate the diagnostic performance of peritoneal endometriosis metabolites.
RESULTS
In the discovery set, peritoneal fluid phosphatidylcholine (34:3) and phenylalanyl-isoleucine were significantly increased in peritoneal endometriosis groups compared with control groups, with AUC 0.77 (95% CI 0.61 to 0.92; P = 0.018) and AUC 0.98 (95% CI 0.95 to 1.02; P < 0.001), respectively. In the validation set, phenylalanyl-isoleucine retained discriminatory performance to distinguish peritoneal endometriosis from controls in both peritoneal fluid (AUC 0.77, 95% CI 0.61 to 0.92; P = 0.006) and serum samples (AUC 0.81, 95% CI 0.64 to 0.99; P = 0.004), with notably stronger discrimination between peritoneal endometriosis and controls in proliferative phase.
CONCLUSION
Our preliminary results propose phenylalanyl-isoleucine as a potential biomarker of peritoneal endometriosis, which may be used as a minimally invasive diagnostic biomarker of peritoneal endometriosis.
Topics: Adult; Ascitic Fluid; Biomarkers; Case-Control Studies; Endometriosis; Female; Humans; Metabolome; Metabolomics; Peritoneal Diseases; Pilot Projects
PubMed: 34446375
DOI: 10.1016/j.rbmo.2021.07.002 -
Journal of Veterinary Emergency and... Jan 2022Interpreting changes in peritoneal fluid helps clinicians manage colic and other diseases in horses. During abdominal problems in the horse, abdominal fluid...
BACKGROUND
Interpreting changes in peritoneal fluid helps clinicians manage colic and other diseases in horses. During abdominal problems in the horse, abdominal fluid characteristics such as color, turbidity, total nucleated and red blood cell counts, cytology, total protein, and l-lactate change in predictable ways, helping the clinician characterize the disease.
DESCRIPTION
Normal abdominal fluid in horses is odorless, clear to light yellow in color, and transparent. Peritoneal fluid becomes more turbid with increasing levels of protein, number of WBCs or RBCs, or with gross contamination following intestinal rupture. The color of abdominal fluid will also change with the type and quantity of cells or other elements present. The transformation of peritoneal fluid color from golden to orange to red represents increasing levels of RBCs, common with strangulating intestinal lesions. Serosanguinous defines fluid that is both turbid and orange to bloody because of increased total protein, WBCs, and RBCs, and is considered classic for diseases characterized by intestinal ischemia. Peritoneal fluid may also be red or blood-colored because of a hemoperitoneum, or secondary to blood contamination during sample collection. l-Lactate measurement in the abdominal fluid has proven invaluable for the identification of strangulating intestinal injury. Cytology acts as an important supplement to cell counts in peritoneal fluid, and the normal ratio of non-degenerate neutrophils:mononuclear cells of 2:1 changes during various gastrointestinal diseases. Culture of peritoneal fluid samples should be performed when septic peritonitis is suspected.
SUMMARY
Abdominal fluid is a sensitive indicator of intestinal injury and a useful tool to direct treatment. Peritoneal fluid evaluation includes gross visual and olfactory examination, nucleated cell count, total protein, RBC count, lactate levels, cytology, and culture. The changes noted in such variables are related to the type and duration of the abdominal problem.
KEY POINTS
Abdominal fluid interpretation has become central to the triage and management of challenging equine colic patients. The transformation of peritoneal fluid color from golden to orange to red represents increasing levels of RBCs, common with strangulating intestinal lesions. Contamination with RBCs at various concentrations may be secondary to vascular (eg, abdominal wall or mesenteric vessels) or splenic trauma during abdominal fluid collection; however, this must be distinguished from orange to red fluid associated with intestinal strangulating obstruction or hemoabdomen Peritoneal fluid analysis reveals abdominal pathology by recognizing specific changes that occur with disease processes affecting the tissues and organs within this cavity. Abdominal fluid examination should be used as a tool to direct treatment rather than the definitive test for diagnosis of the acute abdomen Septic peritonitis in horses most commonly originates secondary to intestinal compromise or accidents (vascular damage, perforation, or surgical manipulation), leading to bacterial translocation into the abdomen.
Topics: Animals; Ascitic Fluid; Colic; Horse Diseases; Horses; Intestinal Obstruction; Peritonitis
PubMed: 35044063
DOI: 10.1111/vec.13117 -
Gynecological Endocrinology : the... Aug 2021The role of leptin in the development of endometriosis has been investigated previously. However, researches on the change of leptin levels in endometriosis remains... (Meta-Analysis)
Meta-Analysis
The role of leptin in the development of endometriosis has been investigated previously. However, researches on the change of leptin levels in endometriosis remains controversial. So, we aimed to clarify changes of leptin levels in patients with endometriosis and their association with the progression of endometriosis. We searched PubMed, Embase, Web of Science, and The Cochrane Library to identify relevant studies published before May 25, 2020. The detected levels of leptin in patients with endometriosis versus controls were evaluated in this meta-analysis. Eighteen studies met our inclusion criteria, five studies detected serum, nine detected peritoneal fluid and another four detected both serum and peritoneal fluid leptin levels. The overall results showed that peritoneal fluid leptin levels in patients with endometriosis was significantly higher than that in the control group, but the serum and corrected peritoneal fluid leptin levels were comparable in both groups. Subgroup analysis failed to eliminate the high degree of heterogeneity included in the studies and showed that peritoneal fluid leptin levels were significantly elevated in both early and advanced endometriosis. In conclusion, peritoneal fluid rather than serum leptin levels was elevated in patients with endometriosis, which did not seem to be related to the severity of endometriosis, but was related to body mass index.
Topics: Ascitic Fluid; Biomarkers; Body Mass Index; Cross-Sectional Studies; Disease Progression; Endometriosis; Female; Humans; Leptin
PubMed: 33355014
DOI: 10.1080/09513590.2020.1862789 -
JCI Insight Jun 2023The omentum contains immune cell structures called milky spots that are niches for transcoelomic metastasis. It is difficult to remove the omentum completely, and there...
The omentum contains immune cell structures called milky spots that are niches for transcoelomic metastasis. It is difficult to remove the omentum completely, and there are no effective strategies to minimize the risk of colonization of preserved omental tissues by cancer cells that circulate in the peritoneal fluid. Normal saline is commonly administered into the peritoneal cavity for diagnostic and intraoperative lavage. Here we show that normal saline, when administered into the peritoneal cavity of mice, is prominently absorbed by the omentum, exfoliates its mesothelium, and induces expression of CX3CL1, the ligand for CX3CR1, within and surrounding the omental vasculature. Studies using CX3CR1-competent and CX3CR1-deficient mice showed that the predominant response in the omentum following saline administration is an accumulation of CX3CR1+ monocytes/macrophages that expand milky spots and promote neoangiogenesis within these niches. Moreover, saline administration promoted the implantation of cancer cells of ovarian and colorectal origin onto the omentum. By contrast, these deleterious effects were not observed following i.p. administration of lactated Ringer's solution. Our findings suggest that normal saline stimulates the receptivity of the omentum for cancer cells and that the risk of colonization can be minimized by using a biocompatible crystalloid for lavage procedures.
Topics: Animals; Mice; Saline Solution; Omentum; Ascitic Fluid; Embryo Implantation; Epithelium
PubMed: 37345662
DOI: 10.1172/jci.insight.167336 -
The Western Journal of Emergency... Mar 2016
Topics: Anti-Bacterial Agents; Ascites; Ascitic Fluid; Female; Hepatitis C; Humans; Liver Cirrhosis; Middle Aged; Paracentesis; Ultrasonography
PubMed: 26973749
DOI: 10.5811/westjem.2016.1.29444 -
Molecular Cancer Nov 2023Knowing the homologous recombination deficiency (HRD) status in advanced epithelial ovarian cancer (EOC) is vital for patient management. HRD is determined by...
BACKGROUND
Knowing the homologous recombination deficiency (HRD) status in advanced epithelial ovarian cancer (EOC) is vital for patient management. HRD is determined by BRCA1/BRCA2 pathogenic variants or genomic instability. However, tumor DNA analysis is inconclusive in 15-19% of cases. Peritoneal fluid, available in > 95% of advanced EOC cases, could serve as an alternative source of cell-free tumor DNA (cftDNA) for HRD testing. Limited data show the feasibility of cancer panel gene testing on ascites cfDNA but no study, to date, has investigated HRD testing.
METHODS
We collected ascites/peritoneal washings from 53 EOC patients (19 from retrospective cohort and 34 from prospective cohort) and performed a Cancer Gene Panel (CGP) using NGS for TP53/HR genes and shallow Whole Genome Sequencing (sWGS) for genomic instability on cfDNA.
RESULTS
cfDNA was detectable in 49 out of 53 patients (92.5%), including those with limited peritoneal fluid. Median cfDNA was 3700 ng/ml, with a turnaround time of 21 days. TP53 pathogenic variants were detected in 86% (42/49) of patients, all with HGSOC. BRCA1 and BRCA2 pathogenic variants were found in 14% (7/49) and 10% (5/49) of cases, respectively. Peritoneal cftDNA showed high sensitivity (97%), specificity (83%), and concordance (95%) with tumor-based TP53 variant detection. NGS CGP on cftDNA identified BRCA2 pathogenic variants in one case where tumor-based testing failed. sWGS on cftDNA provided informative results even when tumor-based genomic instability testing failed.
CONCLUSION
Profiling cftDNA from peritoneal fluid is feasible, providing a significant amount of tumor DNA. This fast and reliable approach enables HRD testing, including BRCA1/2 mutations and genomic instability assessment. HRD testing on cfDNA from peritoneal fluid should be offered to all primary laparoscopy patients.
Topics: Female; Humans; BRCA1 Protein; BRCA2 Protein; Mutation; Ovarian Neoplasms; Circulating Tumor DNA; Homologous Recombination; Ascitic Fluid; Ascites; Prospective Studies; Retrospective Studies; Carcinoma, Ovarian Epithelial; Genomic Instability
PubMed: 37932736
DOI: 10.1186/s12943-023-01864-1 -
International Journal of Molecular... Dec 2022Laparoscopy as a diagnostic tool for patients with suspected endometriosis is associated with several potentially life-threatening complications. Therefore, it is...
Laparoscopy as a diagnostic tool for patients with suspected endometriosis is associated with several potentially life-threatening complications. Therefore, it is imperative to identify reliable, non-invasive biomarkers of the disease. The aim of this study was to analyse the concentrations of fibronectin and type IV collagen in peritoneal fluid and plasma to assess their role as potential biomarkers in the diagnosis of endometriosis. Fibronectin and collagen IV protein levels were assessed by surface plasmon resonance imaging (SPRi) biosensors with the usage of monoclonal antibodies. All patients enrolled in the study were referred for laparoscopy for the diagnosis of infertility or chronic pelvic pain (n = 84). The study group included patients with endometriosis confirmed during surgery (n = 49). The concentration of fibronectin in the plasma (329.3 ± 98.5 mg/L) and peritoneal fluid (26.8 ± 11.1 μg/L) in women with endometriosis was significantly higher than in the control group (251.2 ± 84.0 mg/L, 7.0 ± 5.9 μg/L). Fibronectin levels were independent of endometriosis stage ( = 0.874, = 0.469). No significant differences were observed in collagen IV levels ( = 0.385, = 0.465). The presence of elevated levels of fibronectin may indicate abnormalities in cell-ECM signalling during the course of endometriosis, and may be a potential biomarker for early detection.
Topics: Humans; Female; Endometriosis; Ascitic Fluid; Fibronectins; Collagen Type IV; Biomarkers
PubMed: 36555313
DOI: 10.3390/ijms232415669 -
Iranian Journal of Immunology : IJI Jun 2019Endometriosis is a chronic inflammatory disease with the growth of endometrial cells out of uterus and in the peritoneal cavity. T cell subsets participate in the...
BACKGROUND
Endometriosis is a chronic inflammatory disease with the growth of endometrial cells out of uterus and in the peritoneal cavity. T cell subsets participate in the establishment and progress of the disease by producing different cytokines.
OBJECTIVE
To investigate a group of cytokines related to Th1/Th2/Th17/Treg subsets within both peripheral blood and peritoneal fluid (PF) samples from infertile endometriosis women.
METHODS
Peripheral blood and PF samples were collected from 30 infertile endometriosis and 30 non-endometriosis fertile women during laparoscopy. Concentration of cytokines, including TNF-α, IFN-γ, TGF-β1, IL-4, IL-10, IL-17 and IL-23 were evaluated using ELISA method.
RESULTS
Results indicated that the concentration of IFN-γ within serum was significantly reduced in endometriosis group (p=0.001). Regarding PF cytokines, TGF-β1 was increased in endometriosis group (p=0.030). Furthermore, the ratios of IFN-γ/TGF-β1 and IL-17/IL-23 were significantly different between endometriosis and non-endometriosis women in serum samples (p<0.001 and p<0.01 respectively). The ratios of TNF-α/IL-10 and IL-17/IL-10 were also significantly different regarding PF samples between the two studied groups (p<0.04 and p<0.03 respectively). Finally, significant correlations were observed between the levels of IL-17 and IL-23, inflammatory and anti-inflammatory cytokines, in both samples and serum to PF inflammatory cytokines.
CONCLUSION
Based on the results of the present study, in women with endometriosis, the disturbance of cytokines network might gradually activate the inflammatory responses and tissue repair, resulting in endometriosis development after several years.
Topics: Adult; Ascitic Fluid; Cytokines; Endometriosis; Endometrium; Female; Humans; Infertility, Female; Middle Aged; T-Lymphocyte Subsets; Young Adult
PubMed: 31182689
DOI: 10.22034/IJI.2019.80258