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Current Protocols in Immunology Aug 2015Neutrophils represent the first line of defense against bacterial and fungal pathogens. Indeed, patients with inherited and acquired qualitative and quantitative...
Neutrophils represent the first line of defense against bacterial and fungal pathogens. Indeed, patients with inherited and acquired qualitative and quantitative neutrophil defects are at high risk for developing bacterial and fungal infections and suffering adverse outcomes from these infections. Therefore, research aiming at defining the molecular factors that modulate neutrophil effector function under homeostatic conditions and during infection is essential for devising strategies to augment neutrophil function and improve the outcome of infected individuals. This unit describes a reproducible density gradient centrifugation-based protocol that can be applied in any laboratory to harvest large numbers of highly enriched and highly viable neutrophils from the bone marrow of mice both at the steady state and following infection with Candida albicans as described in UNIT. In another protocol, we also present a method that combines gentle enzymatic tissue digestion with a positive immunomagnetic selection technique or Fluorescence-activated cell sorting (FACS) to harvest highly pure and highly viable preparations of neutrophils directly from mouse tissues such as the kidney, the liver or the spleen. Finally, methods for isolating neutrophils from mouse peritoneal fluid and peripheral blood are included. Mouse neutrophils isolated by these protocols can be used for examining several aspects of cellular function ex vivo including pathogen binding, phagocytosis and killing, neutrophil chemotaxis, oxidative burst, degranulation and cytokine production, and for performing neutrophil adoptive transfer experiments.
Topics: Animals; Ascitic Fluid; Bone Marrow Cells; Cell Separation; Mice; Neutrophils
PubMed: 26237011
DOI: 10.1002/0471142735.im0320s110 -
The Netherlands Journal of Medicine Oct 2016Accumulation of fluid in the peritoneal cavity - ascites - is commonly encountered in clinical practice. Ascites can originate from hepatic, malignant, cardiac, renal,... (Review)
Review
Accumulation of fluid in the peritoneal cavity - ascites - is commonly encountered in clinical practice. Ascites can originate from hepatic, malignant, cardiac, renal, and infectious diseases. This review discusses the current recommended diagnostic approach towards the patient with ascites and summarises future diagnostic targets.
Topics: Ascites; Ascitic Fluid; Culture Techniques; Diagnosis, Differential; Heart Failure; Humans; Laparoscopy; Liver Cirrhosis; Neoplasms; Pancreatic Diseases; Paracentesis; Polymerase Chain Reaction; Practice Guidelines as Topic; Tuberculosis; Ultrasonography
PubMed: 27762220
DOI: No ID Found -
Journal of Veterinary Emergency and... Jan 2022Abdominocentesis is commonly used to evaluate the abdominal cavity of the horse. This technique provides valuable diagnostic information as well as the means to monitor... (Review)
Review
BACKGROUND
Abdominocentesis is commonly used to evaluate the abdominal cavity of the horse. This technique provides valuable diagnostic information as well as the means to monitor patients with abdominal diseases being managed medically and to determine their need for surgical management. Complications are uncommon and include trauma to the gastrointestinal tract or spleen, septic peritonitis, or abdominal wall infection.
PROCEDURES
This review describes the indications, utility, patient preparation, and instructions for performing abdominocentesis as well as possible complications reported in horses. Step-by-step instructions are provided for the two most commonly used abdominocentesis techniques in horses, which include the use of a needle (18 Ga, 3.8 cm [1.5 in]) and a teat cannula (9.5 cm [3.75 in]).
SUMMARY
Peritoneal fluid collection and fluid analysis can be used to confirm diagnosis of intraabdominal pathology including inflammatory, infectious, neoplastic, obstructive, and bowel strangulation, leading to additional diagnostic and therapeutic plans.
KEY POINTS
Abdominocentesis is useful as a diagnostic procedure in horses suffering from colic, diarrhea, weight loss, or other conditions involving the abdominal cavity and is an integral component of diagnostic testing for colic at referral institutions or in the field. Abdominal fluid collection using an 18-Ga, 3.8-cm (1.5-in) needle is recommended for adult horses because the needle is long enough to penetrate the peritoneal cavity. The teat cannula technique is recommended for use in adult horses, foals, and miniature horses to reduce the risk of enterocentesis, even though this procedure is more traumatic than using an 18-Ga, 3.8-cm needle. Ultrasonography of the abdomen is a valuable tool in the assessment of any horse with signs of colic, but it is not essential for performing an abdominocentesis successfully.
Topics: Abdomen; Animals; Ascitic Fluid; Colic; Horse Diseases; Horses; Peritonitis
PubMed: 35044064
DOI: 10.1111/vec.13118 -
Immunology Letters May 2017Interleukin-37 (IL-37) has been identified as a novel anti-inflammatory cytokine. The present study aimed to evaluate the expression of IL-37 in serum and in peritoneal...
Interleukin-37 (IL-37) has been identified as a novel anti-inflammatory cytokine. The present study aimed to evaluate the expression of IL-37 in serum and in peritoneal fluid to determine its clinical significance in endometriosis. Enzyme-linked immunosorbent assay (ELISA) was performed to examine serum IL-37 levels in patients with endometriosis and healthy controls. Peritoneal fluid IL-37 mRNA and NFκB expression were quantified by real-time reverse transcription polymerase chain reaction assays. The association of IL-37 levels with clinical factors and prognosis of endometriosis was analysed. We found that IL-37 levels in PF and in serum were significantly higher in patients with endometriosis compared to women without endometriosis (P=0.0005). IL-37 levels were highly expressed in PF [132.38±34.62pg/mL; P<0.0001] than in serum [74.10±13.49pg/mL] in endometriosis patients. IL-37 mRNA expression contrasted with NFκB mRNA expression in PF from patients with endometriosis. A significant inverse correlation was observed between IL-37 mRNA and NFκB mRNA expression. IL-37 expression correlates with endometriosis severity. The affected NFκB mRNA expression in endometriosis contributed the to exhibited increase of IL-37. The increased levels of IL-37 may dampen NFκB activation in endometriosis patients.
Topics: Adult; Ascitic Fluid; Endometriosis; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interleukin-1; NF-kappa B; RNA, Messenger; Young Adult
PubMed: 28322860
DOI: 10.1016/j.imlet.2017.03.012 -
Human Reproduction (Oxford, England) Apr 2023Are there specific autoantibody profiles in patients with endometriosis that are different from those in controls?
STUDY QUESTION
Are there specific autoantibody profiles in patients with endometriosis that are different from those in controls?
SUMMARY ANSWER
This study did not reveal a significantly higher prevalence of autoantibodies in the studied groups of patients.
WHAT IS KNOWN ALREADY
Various inflammatory factors are postulated to be involved in the pathomechanisms of endometriosis, and a potential link exists with autoimmune diseases, which may also play an important role. As the diagnosis of endometriosis remains invasive, it can only be confirmed using laparoscopy with histopathological examination of tissues. Numerous studies have focused on identifying useful biomarkers to confirm the disease, but without unequivocal effects. Autoantibodies are promising molecules that serve as potential prognostic factors.
STUDY DESIGN, SIZE, DURATION
A multicentre, cross-sectional study was conducted over 18 months (between 2018 and 2019), at eight Departments of Obstetrics and Gynaecology in several cities across Poland on 137 patients undergoing laparoscopic examination for the diagnosis of endometriosis.
PARTICIPANTS/MATERIALS, SETTINGS, METHODS
During laparoscopy, we obtained plasma samples from 137 patients and peritoneal fluid (PF) samples from 98 patients. Patients with autoimmune diseases were excluded from the study. Autoantibody profiling was performed using HuProt v3.1 human proteome microarrays.
MAIN RESULTS AND THE ROLE OF CHANCE
We observed no significant differences in the expression of autoantibodies in the plasma or PF between the endometriosis and control groups. The study revealed that in the PF of women with Stage II endometriosis, compared with other stages, there were significantly higher reactivity signals for ANAPC15 and GABPB1 (adj. P < 0.016 and adj. P < 0.026, respectively; logFC > 1 in both cases). Comparison of the luteal and follicular phases in endometriosis patients revealed that levels of NEIL1 (adj. P < 0.029), MAGEB4 (adj. P < 0.029), and TNIP2 (adj. P < 0.042) autoantibody signals were significantly higher in the luteal phase than in the follicular phase in PF samples of patients with endometriosis. No differences were observed between the two phases of the cycle in plasma or between women with endometriosis and controls. Clustering of PF and plasma samples did not reveal unique autoantibody profiles for endometriosis; however, comparison of PF and plasma in the same patient showed a high degree of concordance.
LIMITATIONS, REASONS FOR CAUTION
Although this study was performed using the highest-throughput protein array available, it does not cover the entire human proteome and cannot be used to study potentially promising post-translational modifications. Autoantibody levels depend on numerous factors, such as infections; therefore the autoantibody tests should be repeated for more objective results.
WIDER IMPLICATIONS OF THE FINDINGS
Although endometriosis has been linked to different autoimmune diseases, it is unlikely that autoimmune responses mediated by specific autoantibodies play a pivotal role in the pathogenesis of this inflammatory disease. Our study shows that in searching for biomarkers of endometriosis, it may be more efficient to use higher-throughput proteomic microarrays, which may allow the detection of potentially new biomarkers. Only research on such a scale, and possibly with different technologies, can help discover biomarkers that will change the method of endometriosis diagnosis.
STUDY FUNDING/COMPETING INTEREST(S)
This study was funded by a grant from the Polish Ministry of Health (grant no. 6/6/4/1/NPZ/2017/1210/1352). It was also funded by the Estonian Research Council (grant PRG1076) and the Horizon 2020 Innovation Grant (ERIN; grant no. EU952516), Enterprise Estonia (grant no. EU48695), and MSCA-RISE-2020 project TRENDO (grant no. 101008193). The authors declare that there is no conflict of interest.
TRIAL REGISTRATION NUMBER
N/A.
Topics: Humans; Female; Endometriosis; Ascitic Fluid; Autoantibodies; Cross-Sectional Studies; Proteome; Proteomics; Biomarkers; Autoimmune Diseases; Adaptor Proteins, Signal Transducing; DNA Glycosylases
PubMed: 36749097
DOI: 10.1093/humrep/dead011 -
Methods in Molecular Biology (Clifton,... 2022The accumulation of peritoneal fluid, referred to as ascites, is common in ovarian cancer. This fluid is a complex mixture that may include cells as well as a diverse...
The accumulation of peritoneal fluid, referred to as ascites, is common in ovarian cancer. This fluid is a complex mixture that may include cells as well as a diverse array of cytokines and growth factors. Here we describe a comprehensive method to process ascites to maximize data collection. The cellular fraction and fluid are first separated by centrifugation. The fluid can be frozen for later analysis of soluble factors or for use in in vitro experiments. The cellular fraction can be processed to analyze its composition or stored for future use.
Topics: Ascites; Ascitic Fluid; Cytokines; Female; Humans; Intercellular Signaling Peptides and Proteins; Ovarian Neoplasms
PubMed: 34918288
DOI: 10.1007/978-1-0716-1956-8_5 -
The Veterinary Clinics of North... Jan 2017Canine peritoneal fluid analysis results were retrospectively reviewed to assess the appropriateness of different classification schemes. Cutoffs of 3000 cells/μL and... (Review)
Review
Canine peritoneal fluid analysis results were retrospectively reviewed to assess the appropriateness of different classification schemes. Cutoffs of 3000 cells/μL and 2.5 g/dL protein are recommended. Analyzing the total nucleated cell count and total protein concentration is only the first step in peritoneal fluid analysis; microscopic examination, clinical presentation, and other laboratory data are all important in determining the final classification of peritoneal fluid analysis, keeping in mind that the most important aspect of fluid analysis is not what something is called, but whether it helps achieve a diagnosis. Discussion of effusion mechanisms, study observations, and recommended diagnostic steps after fluid analysis are included.
Topics: Animals; Ascitic Fluid; Cytological Techniques; Dog Diseases; Dogs
PubMed: 27531039
DOI: 10.1016/j.cvsm.2016.07.008 -
International Journal of Medical... 2022Interleukin-17 (IL-17) is known as a Th17-cell-derived proinflammatory cytokine, which plays a pivotal role in several inflammatory and autoimmune diseases such as... (Review)
Review
Interleukin-17 (IL-17) is known as a Th17-cell-derived proinflammatory cytokine, which plays a pivotal role in several inflammatory and autoimmune diseases such as systemic lupus erythematosus (SLE), rheumatoid arthritis, and psoriasis. Emerging evidence has shown that IL-17 is linked to endometriosis, although the etiology of endometriosis is still unknown. The IL-17 expression is up-regulated in serum, peritoneal fluid (PF) and endometriotic lesions from patients with endometriosis but the related regulation mechanisms are complex and obscure. Meanwhile, the specific roles of IL-17 in endometriosis are also worthy of further exploration. Through the integration and summary of literature, we conclude that the secretion of IL-17 increases under the regulation of ectopic microenvironment and other factors, and then IL-17 is deeply involved in endometriosis in the regulation of immune microenvironment, the invasion and growth of ectopic lesions, and so on, which implies its therapeutic value in this disorder.
Topics: Ascitic Fluid; Endometriosis; Female; Humans; Interleukin-17; Th17 Cells
PubMed: 35582411
DOI: 10.7150/ijms.71972 -
International Journal of Surgery... Nov 2023Staging laparoscopy for gastric cancer is recommended to assess the tumor's locoregional extension and exclude peritoneal disease. As there is no consensus on optimizing...
BACKGROUND
Staging laparoscopy for gastric cancer is recommended to assess the tumor's locoregional extension and exclude peritoneal disease. As there is no consensus on optimizing the procedure's diagnostic accuracy, we aimed to systematically review the literature on operative techniques, followed by peritoneal lavage fluid assessment in gastric cancer patients. Specifically, we sought to indicate the most common characteristics of the procedure and cytological evaluation.
METHODS
This study was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA). The protocol for this systematic review was registered on PROSPERO database (CRD: 42022306746). On September 2022, a search was carried out using Embase, Medline ALL, Cochrane Central Register of Controlled Trials, and Web of Science Core Collection.
RESULTS
The search identified 1632 studies on staging laparoscopy and 2190 studies on peritoneal fluid assessment. Some 212 studies were included. Open Hasson was the method of choice in accessing the peritoneal cavity in 65% of the studies, followed by establishing a pneumoperitoneum at 10-12 mmHg in 52% of reports. Most frequently, the patient was positioned supine (70%), while a 30° scope and three ports were used to assess the peritoneal cavity clockwise (72%, 77%, and 85%, respectively). Right and left upper abdomen quadrants were the predominant area of laparoscopic exploration (both 65%), followed by the primary tumor region (54%), liver and pelvis (both 30%), and small bowel and spleen (19% and 17%, respectively). Regions of peritoneal lavage and aspiration were limited to the pelvis (50%), followed by right and left upper abdomen quadrants (37.5% and 50%, respectively). No studies compared different methods of operative techniques or analysis of ascites/fluid.
CONCLUSIONS
This study indicates a high heterogeneity in the technique of staging laparoscopy and peritoneal fluid assessment in gastric cancer patients. Further research and initiatives to reach a consensus on the standardization of the procedure are warranted.
Topics: Humans; Stomach Neoplasms; Ascitic Fluid; Neoplasm Staging; Laparoscopy; Peritoneal Lavage
PubMed: 37581636
DOI: 10.1097/JS9.0000000000000632 -
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