Peritoneal Fluid Of Patients Research Articles

Effect Of Vitamin D Administration On Interleukin 6 (IL-6) Levels In Peritoneal Fluid In Endometrioma Patients

BACKGROUND: An endometrioma is a cyst that occurs when endometrial tissue grows on the epithelium of the ovary. Vitamin D is a steroid hormone that can be extracted from commonly available foods and can be synthesized by humans when exposed to sunlight. Vitamin D deficiency has been reported to be associated with various pathologies, including endometrioma. Vitamin D has been reported to have anti-angiogenic effects that may inhibit the growth of endometriotic implants. AIM: Proving the effect of vitamin D administration on interleukin 6 (IL-6) levels in peritoneal fluid in patients with endometrioma. METHOD: Experimental research with two group, post-test only design with single blind that carried out at Central General Hospital (RSUP) Dr. Kariadi Semarang. The research subject was 50 patients with endometrioma on ovary. The definite diagnosis of endometrioma is made based on histopathological examination of tissue samples after the patient undergoes surgery. Patients will be divided into 2 groups, namely the vitamin D3 supplementation group and placebo. The intervention was carried out by providing vitamin D3 50,000 IU/week for a month. Evaluation of serum vitamin D levels were obtained using venous blood samples immediately when the patient was undergoing surgery. Interlukin-6 levels were obtained using peritoneal fluid samples obtained when the patient underwent surgery. Statistical analysis was carried out using the Independent T Test, Mann-Whitney U, Dependent T Test and Wilcoxon. Results are significant if p <0.05. Statistical analysis was carried out using SPSS edition 26. RESULT: Peritoneal fluid interleukin-6 (IL-6) levels between the vitamin D supplementation group and the placebo group did not show a significant difference (p=0.554). CONCLUSION: Vitamin D supplementation in endometrioma patients is not associated with peritoneal fluid interleukin-6 (IL-6) levels.

Retinoic acid receptor alpha expression in endometrioid heterotopias and the endometrium of patients with endometriosis

BACKGROUND: Vitamin A is a fat-soluble compound, which is a biologically inactive food form and undergoes transformation by cytosolic alcohol dehydrogenases and microsomal retinol dehydrogenases into retinaldehyde, followed by oxidation by retinaldehyde dehydrogenases 1, 2 and 3 into retinoic acid. All-trans-retinoic acid has been identified as an active metabolite of retinoic acid, indirectly participating in various processes: differentiation, proliferation and apoptosis. Based on a number of studies, the role of retinoic acid and its receptors in various diseases has been proven. For example, the destruction of the RAR-β gene by the integration of the hepatitis B virus into its loci can be a factor in the development of human hepatocellular carcinoma. Subsequently, it was described that the characteristic point of interruption of translocation t(15:17), observed in acute promyelocytic leukemia, was located at the locus encoding retinoic acid receptor-α. As a result, in patients with acute promyelocytic leukemia, complete remission of the disease could often be achieved by treatment with high doses of all-trans-retinoic acid. Existing data on the ability of retinol to influence the processes of differentiation, proliferation, and apoptosis indicate that retinol may be involved in the pathogenesis of endometriosis, but the mechanisms of its effect on the disease are subject to further study. AIM: The aim of this study was to evaluate the retinoic acid receptor alpha expression in endometrioid heterotopias and the endometrium of patients with different endometriosis stages according to the Revised American Society for Reproductive Medicine classification. MATERIALS AND METHODS: This was a prospective study to determine the retinoic acid receptor alpha expression in the endometrium and endometrioid heterotopias of patients with endometriosis (n = 28) and of the control group (n = 10). RESULTS: The retinoic acid receptor alpha expression in endometrioid heterotopias was lower compared to the eutopic endometrium of patients with endometriosis and the endometrium of patients in the control group by 3.4 and two times, respectively (p 0.001; the Kruskal – Wallis method). CONCLUSIONS: The data obtained indicate the involvement of retinol in the pathogenesis of endometriosis. Despite the relative increase in retinol level in the peripheral blood and peritoneal fluid in patients with endometriosis compared to the control group, shown in our previous studies, the reduced retinoic acid receptor alpha expression in endometrioid heterotopias does not allow for full binding of vitamin A to the receptor and is accompanied by its improper metabolism.

Downregulation of pattern recognition receptors on macrophages involved in aggravation of endometriosis.

In women of reproductive age, endometriosis may contribute to dysmenorrhea, chronic pelvic pain, dyspareunia, infertility, adenomyosis, and endometrial ovarian cyst (EOC). Recent studies have shown that chronic inflammation occurs in the pelvis of endometriosis patients and that this inflammation is exacerbated by immunosuppression, leading to survival endometrial debris. However, the detailed immunological mechanisms underlying the aggravation of inflammation and immunosuppression in endometriosis patients remain unclear. We investigate the alarmins (high-mobility group box-1, IL-33, IL-1α, and S100B protein), proinflammatory cytokines (IL-6 and IL-1β), and immune cells (CD8+ T cells, CD4+ T cells, natural killer cells, natural killer T cells, dendritic cells, and macrophages) in peritoneal fluid of patients with EOC using enzyme-linked immunosorbent assay, electrochemiluminescence, and flow cytometry. Then, we analyzed the correlation between these factors and the aggravating indicators of endometriosis, tumor size and revised American Society for Reproductive Medicine (r-ASRM) score. Unexpectedly, there was no correlation between each alarmin level and aggravating indicators. However, the expression of pattern recognition receptors, toll-like receptor 4, and receptor of advanced glycation end-products on macrophages was inversely correlated with aggravating indicators. The aggravation of endometriosis is associated with a decrease in alarmin receptors but not alarmin levels. Investigation of innate immune systems, such as alarmins and their receptors, may help elucidate new mechanisms of endometriosis.

A higher number of exhausted local PD1+, but not TIM3+, NK cells in advanced endometriosis

Endometriosis (EMT) is a chronic inflammatory disease characterized by the presence and growth of endometrial-like glandular epithelial and stromal cells outside the uterus. Natural Killer (NK) cell dysfunction/exhaustion has been shown in patients with EMT. In this case-control study, we compared the frequency of exhausted PD-1 or TIM-3 positive NK cells in peripheral blood (PB) and peritoneal fluid (PF) of women with advanced endometriosis to control fertile women. PB and PF were collected from women aged 25–40 who underwent the laparoscopic procedure, including 13 stages III/IV endometriosis and 13 control samples. Multicolor flowcytometry was used to compare the frequency of PD-1 or TIM-3 positive NK (CD3−CD56+) cells in PB and PF of two groups. We demonstrated a higher percentage of PD-1+ NK cells in the peritoneal fluid of patients with endometriosis rather than controls (P-value = 0.039). This significance was related to stage IV of endometriosis (P-value = 0.047). We can not show any significant difference in the number of PD-1 or TIM-3 positive NK cells in peripheral blood. Our results suggest a local exhausted NK cell response in endometriosis that can be a leading factor in the endometriosis pathogenesis.

Focused Assessment with Sonography for Trauma (FAST) Exam: Image Acquisition.

Over the past twenty years, the Focused Assessment with Sonography for Trauma (FAST) exam has transformed the care of patients presenting with a combination of trauma (blunt or penetrating) and hypotension. In these hemodynamically unstable trauma patients, the FAST exam permits rapid and noninvasive screening for free pericardial or peritoneal fluid, the latter of which implicates intra-abdominal injury as a likely contributor to the hypotension and justifies emergent abdominal surgical exploration. Further, the abdominal portion of the FAST exam can also be used outside of the trauma setting to screen for free peritoneal fluid in patients who become hemodynamically unstable in any context, including after procedures that may inadvertently injure abdominal organs. These "non-trauma" situations of hemodynamic instability are often triaged by providers from specialties other than emergency medicine or trauma surgery who are not familiar with the FAST exam. Therefore, there is a need to promulgate knowledge about the FAST exam to all clinicians caring for critically ill patients. Toward this end, this article describes FAST exam image acquisition: patient positioning, transducer selection, image optimization, and exam limitations. Since the free fluid is likely to be found in specific anatomic locations that are unique for each canonical FAST exam view, this work centers on the unique image acquisition considerations for each window: subcostal, right upper quadrant, left upper quadrant, and pelvis.

Evaluation of Proteasome and Immunoproteasome Levels in Plasma and Peritoneal Fluid in Patients with Endometriosis.

Endometriosis is a chronic disease in which the endometrium cells are located outside the uterine cavity. The aim of this study was to evaluate circulating 20S proteasome and 20S immunoproteasome levels in plasma and peritoneal fluid in women with and without endometriosis in order to assess their usefulness as biomarkers of disease. Concentrations were measured using surface plasmon resonance imaging biosensors. Patients with suspected endometriosis were included in the study-plasma was collected in 112 cases and peritoneal fluid in 75. Based on the presence of endometriosis lesions detected during laparoscopy, patients were divided into a study group (confirmed endometriosis) and a control group (patients without endometriosis). Proteasome and immunoproteasome levels in both the plasma (p = 0.174; p = 0.696, respectively) and the peritoneal fluid (p = 0.909; p = 0.284, respectively) did not differ between those groups. There was a statistically significant difference in the plasma proteasome levels between patients in the control group and those with mild (Stage I and II) endometriosis (p = 0.047) and in the plasma immunoproteasome levels in patients with ovarian cysts compared to those without (p = 0.017). The results of our study do not support the relevance of proteasome and immunoproteasome determination as biomarkers of the disease but suggest a potentially active role in the pathogenesis of endometriosis.

Proinflammatory cytokines in peritoneal fluid in women with peritoneal adhesions

The objective: to investigate the level of pro-inflammatory cytokines in the peritoneal fluid in women of reproductive age with peritoneal adhesions. Materials and methods. 58 patients verified during laparoscopy were included in the study: of them, 45 patients with stages I–II of the adhesion process, and 13 – with stages III–IV of the spread of the process. The control group consisted of 30 women of reproductive age with intact reproductive function without adhesions in the pelvis, who underwent laparoscopic sterilization. The content of IL-1, I IL-2, IL-6, and tumor necrosis factor-alpha in the peritoneal fluid was analyzed. Results. Studies of the content of pro-inflammatory cytokines IL-1 (192.5±34.8 pg/ml and268.4±48.3 pg/ml), IL-2 (145.9±21.3 pg/ml and 234.8±42.5 pg/ml), IL-6 (118.7±8.4 pg/ml and698.4±22.6 pg/ml) and TNF-α (10.2±4.8 pg/ml and 29.6±2.8 pg/ml) in peritoneal fluid showed that their levels were significantly higher (p<0.01) in women with widespread stages of the adhesion process and the lowest in CG – 18.2±4.2 pg, respectively/ml, 22.4±4.3 pg/ml and 26.4±4.7 pg/ml. The level of IL-6 – 698.4±22.6 pg/ml in the peritoneal fluid of patients with III–IV stages of the adhesion process was significantly higher (p<0.01) than the level of IL-6 – 118.7±8.4 pg/ml in the peritoneal fluid of women with stage I–II spread of adhesions and in the control group – 26.4±4.7 pg/ml. Сonclusions. With the increase in the prevalence of the adhesion process, there is a significant reliable increase in the content of cytokines in the peritoneal fluid, while the peritoneal fluid plays a significant role in its formation.

Investigation of the Changes in Concentrations of Vitamin D-Binding Protein and Lactoferin in Plasma and Peritoneal Fluid of Patients with Endometriosis.

An evaluation of the association between the concentrations of vitamin D-binding protein and lactoferrin in the plasma and peritoneal fluid may facilitate the elucidation of molecular mechanisms in endometriosis. Vitamin D-binding protein and lactoferrin concentrations were measured by ELISA in plasma and peritoneal fluid samples from 95 women with suspected endometriosis as classified by laparoscopy into groups with (n = 59) and without endometriosis (n = 36). There were no differences (p > 0.05) in the plasma and peritoneal fluid concentrations of vitamin D-binding protein and lactoferrin between women with and without endometriosis. In women with endometriosis, there was a significant correlation between plasma and peritoneal fluid vitamin D-binding protein concentrations (r = 0.821; p = 0.000), but there was no correlation between lactoferrin concentrations in those compartments (r = 0.049; p > 0.05). Furthermore, in endometriosis, lactoferrin was found to correlate poorly with vitamin D-binding protein (r= -0.236; p > 0.05) in plasma, while in the peritoneal fluid, the correlation between those proteins was significant (r = 0.399; p = 0.002). The characteristic properties of vitamin D-binding protein and lactoferrin and the associations between their plasma and peritoneal fluid concentrations found in women with endometriosis may provide a novel panel of markers to identify high-risk patients in need of further diagnostic measures.

Прозапальні цитокіни в перитонеальній рідині жінок з перитонеальними спайками

Purpose - to investigate the level of pro-inflammatory cytokines in the peritoneal fluid in women of reproductive age with peritoneal adhesions. Materials and methods. 58 patients (the main group) verified during laparoscopy were included in the study: of them, 45 patients with stages I-II of the adhesion process, 13 - with stages III-IV of the spread of the process. The control group (CG) consisted of 30 women of reproductive age with intact reproductive function without adhesions in the pelvis, who underwent laparoscopic sterilization. The content of interleukin (IL) 1, IL-2, IL-6 and tumor necrosis factor alpha (TNF-α) in the peritoneal fluid was analyzed. Results. The study of the content of proinflammatory cytokines in peritoneal fluid showed that their levels were significantly higher (p<0.01) in women with stage I-II and stage III-IV adhesions: IL-1 - 192.5±34.8 pg/ml and 268.4±48.3 pg/ml, respectively; IL-2 - 145.9±21.3 pg/ml and 234.8±42.5 pg/ml, respectively; IL-6 - 118.7±8.4 pg/ml and 698.4±22.6 pg/ml, respectively; TNF-α - 10.2±4.8 pg/ml and 29.6±2.8 pg/ml, respectively. These levels were the lowest in CG women: IL-1 - 18.2±4.2 pg/ml; IL-2 - 22.4±4.3 pg/ml; IL-6 - 26.4±4.7 pg/ml; and TNF-α was not detected. The level of IL-6 in peritoneal fluid of patients with stages III-IV of the adhesive process (698.4±22.6 pg/ml) was significantly higher (p<0.01) than that in women with stages I-II of adhesion prevalence (118.7±8.4 pg/ml) and in women of the CG (26.4±4.7 pg/ml). Conclusions. With an increase in the prevalence of the adhesive process, the content of cytokines in the peritoneal fluid increases significantly, and the peritoneal fluid plays a significant role in its formation. The research was carried out in accordance with the principles of the Helsinki Declaration. The study protocol was approved by the Local Ethics Committee of the participating institution. The informed consent of the patient was obtained for conducting the studies. No conflict of interests was declared by the author.

Autoantibody screening of plasma and peritoneal fluid of patients with endometriosis.

Are there specific autoantibody profiles in patients with endometriosis that are different from those in controls? This study did not reveal a significantly higher prevalence of autoantibodies in the studied groups of patients. 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. 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. 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. 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. 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. 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. 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. N/A.

Survivin expression at the mRNA level in tumors and the protein concentration in the serum and peritoneal fluid in patients with serous ovarian tumors.

Ovarian cancer is one of the gynecological cancers that have the worst prognosis. The expression of the proteins from the IAP family (inhibitor of apoptosis protein), including survivin, is observed in many types of cancer. The aim of the study was to evaluate survivin at the mRNA level in tumors and the protein concentration in the serum and peritoneal fluid of patients with serous ovarian cancer in order to assess the relationship between the concentration of survivin and the histological subtypes of cancer. The study group consisted of 55 women, including patients with serous ovarian cancer (n = 30, nine low-grade serous carcinoma LGSC, 21 high-grade serous carcinoma HGSC), serous cysts (n = 10) and the control group (n = 15). The concentration of protein in the peritoneal fluid and serum was assessed using ELISA tests. The expression of survivin gene BIRC5 in the tumors was assessed using the RT-qPCR method. The data that was obtained indicated that the concentration of survivin was higher in the serum of the women with serous ovarian cancer compared those that had benign tumors (p < 0.05) and the control group (p < 0.001). The survivin concentration was also higher in both the serum and peritoneal fluid in the HGSC group compared to the LGSC group (p < 0.001). The mRNA level was highest in the HGSC group, and there was a statistically significant difference compared to those in the benign tumor group and HGSC group ( p < 0.05). The observed changes prove that the expression level increases significantly in HGSC in both the protein and mRNA levels. Based on these findings, it can be assumed that assessing this parameter could be a useful additional indicator of the progression and differentiation of this type of cancer. However, this requires further research in a larger group of patients and possibly in other types of ovarian cancer.

Meta-analyses reveal serum or plasma Interleukin-6 as a biomarker for malignant ovarian neoplasia.

Interleukin-6 (IL-6) has been implicated in various malignancies, including ovarian cancer. However, mixed results have been observed regarding IL-6 levels in different ovarian conditions. This meta-analysis was performed to determine IL-6 levels in the peritoneal fluid and peripheral blood among patients with various adnexal masses. Most popular English databases were searched using a predefined search formula. All studies comparing IL-6 levels in plasma, serum or peritoneal fluid of patients with benign tumors, ovarian neoplasms, and healthy controls were included based on inclusion and exclusion criteria. 5953 patients from 22 primary publications raging from 1994 to 2021 were included in the meta-analyses. A pooled IL-6 Mean Difference (MD) of 41pg/mL for malignant tumors compared to benign ones, with a Confidence Interval (CI) between 19.8 and 62.2, a Z-score of 3.79, and statistical significance with a p=0.0002 was observed. Pooled results for healthy versus benign ovarian conditions showed an MD of 5.45pg/mL for serum or plasma IL-6 measurements in favor of benign tumors (CI:0.66-10.25, Z=2.23 and p=0.03). The analysis showed an MD for IL-6 levels of 19.59pg/mL for healthy controls versus malignant ovarian tumors. Peritoneal fluid measurements regarding IL-6's levels showed no significant difference between benign or malignant masses. Higher levels of plasma or serum IL-6 in ovarian neoplasia patients compared to benign conditions or healthy controls identify IL-6 as a discerning factor between benign or malignant ovarian tumors and a potential biomarker for ovarian malignancy.

Plasma and Peritoneal Poly (ADP-Ribose) Polymerase Levels in Patients with Endometriosis.

The evidence of poly (ADP-ribose) polymerase (PARP) association with the immune response could be coherent with the immunological theory of endometriosis and suggests the possibility of a new research direction. The aim of the study was to evaluate the levels of PARP in plasma and peritoneal fluid of patients with and without endometriosis. It was a multicenter, cross-sectional study. Plasma and peritoneal fluid samples were collected from patients with and without endometriosis during planned laparoscopic procedures in eight clinical centers. In total, 84 samples of plasma and 84 samples of the peritoneal fluid were included in the final analyses. Double-antibody sandwich enzyme-linked immunosorbent assay was performed in order to assess levels of PARP in collected samples. No statistically significant differences regarding the detected levels of PARP in plasma and peritoneal fluid comparing patients with and without endometriosis were observed. Patients with a history of infertility had significantly higher plasma PARP concentrations (p = 0.04). We have not observed the potential role of PARP concentration levels in plasma nor peritoneal fluid as an endometriosis biomarker. We have determined an association between a higher plasma PARP concentration and a history of infertility.

CTLA4-Linked Autoimmunity in the Pathogenesis of Endometriosis and Related Infertility: A Systematic Review.

Several studies, although with conflicting results, have sought to determine the concentration of soluble CTLA4 antigens in peripheral blood plasma and peritoneal fluid in patients with endometriosis-related infertility. A systematic review was performed according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) through a search of the following databases: MEDLINE, EMBASE, Global Health, The Cochrane Library, Health Technology Assessment Database and Web of Science, and Clinical Trials research register. We included observational or prospective human and animal studies with any features related to endometriosis and/or infertility studies involving CTLA4-related pathogenesis published in English. The results of studies in which the size and characteristics of the observed groups were not stated were excluded. From the initial pool of 73 publications identified and screened, we finally included 5 articles to summarize the most recent knowledge about CTLA4-linked autoimmunity in the pathogenesis of endometriosis and related infertility. Evidence from clinical studies shows that CTLA4-based autoimmunity is involved in the maintenance of chronic inflammation in the peritoneal environment, with pre-clinical evidence of anti-CTLA antibodies as a potential novel target therapy for endometriosis. However, CTLA4 gene analyses do not support findings of CTLA4-linked autoimmunity as a primary determinant of the pathogenesis of endometriosis. These findings underlie the role of complex interactions within the family of immune checkpoint molecules involved. Further studies are needed to investigate the clinical relevance of anti-CTLA target therapy, taking into account the potential adverse events and repercussions of novel immunologic therapy modalities. However, with the general scarcity of studies investigating this topic, the clinical importance of CTLA4 autoimmunity still remains unclear.

Analysis of CXCL8 and its receptors CXCR1/CXCR2 at the mRNA level in neoplastic tissue, as well as in serum and peritoneal fluid in patients with ovarian cance.

Understanding the relationship between the coexistence of inflammatory and neoplastic processes in ovarian cancer, particularly those involving chemokines and their receptors, may help to elucidate the involvement of the studied parameters in tumor pathogenesis and could lead to improved clinical applications. Therefore, the present study aimed to analyze the levels of C-X-C motif chemokine ligand 8 (CXCL8), and its receptors C-X-C chemokine receptor (CXCR)1 and CXCR2, in the serum and peritoneal fluid of women with ovarian cancer, and to evaluate the association between the expression of these parameters in tumor tissue and patient characteristics, particularly the degree of histological differentiation. The study group included women with ovarian cancer diagnosed with serous cystadenocarcinoma International Federation of Gynecology and Obstetrics stage IIIc and a control group, which consisted of women who were diagnosed with a benign lesion (serous cystadenoma). The transcript levels of CXCL8, CXCR1 and CXCR2 were evaluated using reverse transcription-quantitative PCR (RT-qPCR). The quantitative analysis was carried out using the LightCycler® 480 System and GoTaq® 1-Step RT-qPCR System, according to the manufacturers' instructions. The concentration of CXCL8 in serum and peritoneal fluid was determined using a Human Interleukin-8 ELISA kit, and the concentrations of CXCR1 and CXCR2 were determined using the CLOUD-CLONE ELISA kit. Local and systemic disturbances in immune and inflammatory responses involving the CXCL8 chemokine and its receptors indicated the involvement of these studied parameters in the pathogenesis of ovarian cancer. Immunoregulation of the CXCL8-CXCR1 system may influence the course of the inflammatory process accompanying ovarian cancer development, which may result in the identification of novel clinical applications; however, further studies are required.

Abstract 6144: Novel M2 macrophage gene signature as a biomarker of poor outcomes independent of TNM stage in peritoneal liquid biopsy of gastric cancer patients

Abstract Introduction: The infiltration or density of immunosuppressive or M2-macrophages has been correlated with worse prognosis in gastric cancer (GC). We hypothesized that a highly polarized transcriptional phenotype of M2 macrophages would be prognostic if detected in primary and peritoneal fluid samples of GC. Methods: We derived a novel transcriptomic gene signature (MRC1, MS4A4A, CD36, CCL13, CCL18, CCL23, SLC38A6, FGL2, FN1, MAF) from published highly differentially expressed M2 genes that were identified from peripheral blood. We also evaluated a M1 signature (CCR7, IL2RA, CXCL11, CCL19, CXCL10, PLA1A, PTX3) and T cell cytolytic signature (GZMA, GZMB, GZMH, GZMM, PRF1). In primary GC, we used the UCSC Xena tool to analyze the TCGA Stomach Cancer cohort and compared transcriptional signature expression to histologic subtype and pathologic stage. We used log-rank test to compare Kaplan-Meier curves of overall survival (OS) and disease-free interval (DFI) for high versus low signature expression. In our institutional cohort consisting of GC patients with a range of T and N stages as well as with and without metastatic disease (n=28), we evaluated peritoneal fluid samples using single cell RNA sequencing for high and low signature expression in CD68+CD163+ monocytes and used log-rank test to compare OS. Results: A total of 341 TCGA entries of primary GC were included. High expression of the M2 signature in GC was associated with diffuse- and mucinous-type adenocarcinoma, compared to tubular- and papillary-type adenocarcinoma (p&amp;lt;0.001). High M2 signature expression was associated with later stage (Stage II or above) compared to early-stage GC (stage I) (p=0.02). Low M2 signature was associated with significantly improved 5-year OS (p=0.03) and DFI (p=0.04). M1 signature and T cell cytolytic signature were not associated with stage, OS, or DFI in primary GC. In GC peritoneal fluid samples, we identified patients with high (n=15) and low (n=13) signature expression in M2 cells. Independent of TNM-stage, high M2 signature expression in CD68+CD163+ cells within the peritoneal fluid of GC patients was associated with worse OS (median OS not reached at 30 months for low signature expression and 5.5 months for high expression, hazard ratio 2.82, p=0.048). Conclusions: Our study validates a unique M2-associated transcriptomic gene signature as a biomarker of poor outcomes in both primary gastric tumors and peritoneal fluid of GC patients. Because both endoscopic biopsy and laparoscopy with peritoneal fluid collection are part of the standard workup for locoregional GC, an ubiquitous immune signature detected in primary tumor and peritoneal fluid of patients across TNM stages may serve as a clinically valuable indicator of an immunosuppressive patient state associated with poor survival. Citation Format: Kevin M. Sullivan, Haiqing Li, Kelly Mahuron, Yate-Ching Yuan, Joseph Chao, Benjamin Paz, Haiyong Han, Yuman Fong, Yanghee Woo. Novel M2 macrophage gene signature as a biomarker of poor outcomes independent of TNM stage in peritoneal liquid biopsy of gastric cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6144.

Whole-Exome Sequencing Reveals Clinical Potential of Circulating Tumor DNA from Peritoneal Fluid and Plasma in Endometrial Cancer.

Simple SummaryStudies on biomarkers for endometrial cancer using circulating tumor DNA (ctDNA) from liquid biopsy samples are lacking. The aim of this study was to analyze gDNA from tumor tissues and ctDNA derived from peritoneal fluid and plasma samples and determine their mutational concordance via microsatellite instability, copy number alteration, and mutational signature analyses using whole-exome sequencing and P53 immunohistochemistry. ctDNA of two patients with negative cytology presented TP53 mutations concordant with those in the tissue, and the ctDNA of a patient with positive cytology harbored both TP53 and POLE mutations, although none were detected in the tissue. This study is the first to demonstrate comprehensive genomic concordance between ctDNA from the peritoneal fluid and tumor gDNA in endometrial cancer at the whole-exome level.Endometrial cancer (EC) is the most common type of gynecological cancer. Studies comparing tumor gDNA and ctDNA isolated from the plasma and peritoneal fluid of EC patients are limited. Whole-exome sequencing and P53 immunohistochemistry of 24 paired tissue, plasma, and peritoneal fluid samples from 10 EC patients were performed to analyze somatic mutations, copy number alterations, microsatellite instability, and mutational signatures. Mutations in cancer-related genes (KMT2C, NOTCH2, PRKAR1A, SDHA, and USP6) and genes related to EC (ARID1A, CTNNB1, PIK3CA, and PTEN) were identified with high frequencies among the three samples. TP53 and POLE mutations, which are highly related to the molecular classification of EC, were identified based on several key observations. The ctDNA of two patients with negative peritoneal fluid presented TP53 mutations concordant with those in tissues. ctDNA from the plasma and peritoneal fluid of a patient with positive cytology harbored both TP53 and POLE mutations, although none were detected in tissues. Additionally, the patient presented with wild type P53 immunohistochemistry, with a focal “high” expression in a “low” wild type background. The tissues and peritoneal fluid of 75% EC patients showed concordant microsatellite instability. Furthermore, we observed strong mutational concordance between the peritoneal fluid and tumors. Our data suggest that the ctDNA from peritoneal fluid might be a suitable biomarker for identifying the mutational landscape of EC and could complement tumor heterogeneity.

Positive peritoneal swab in SARS-CoV-2 patients undergoing abdominal emergency surgery: effect or cause?

PurposeThe presence of the SARS-CoV-2 in the peritoneal fluid is a matter of debate in the COVID-19 literature. The study aimed to report the prevalence of SARS-CoV-2 in the peritoneal fluid of patients with nasopharyngeal swab tested positive for SARS-CoV-2 undergoing emergency surgery and review the literature.MethodsThe present study was conducted between March 2020 and June 2021. Diagnosis of SARS-CoV-2 positivity was confirmed by preoperative real-time reverse transcriptase-polymerase chain reaction (RT-PCR).ResultsEighteen patients with positive nasopharyngeal swabs were operated in emergency in two third-level Italian hospitals. In 13 of these patients (72%), a peritoneal swab was analyzed: SARS-CoV-2 RNA was found in the abdominal fluid of two patients (15%). Neither of them had visceral perforation and one patient died. In ten patients with negative peritoneal swabs, visceral perforation and mortality rates were 30% and 20%, respectively.ConclusionSARS-CoV-2 peritoneal positivity is rare. Abdominal surgery can, therefore, be safely performed in patients with COVID-19 using standard precautions. The correlation with a visceral perforation is not evaluable. The clinical outcomes seem uninfluenced by the viral colonization of the peritoneum. Assessment in large series to provide definitive answers about the involvement of the SARS-CoV-2 in the peritoneum will be challenging to coordinate.

CD39 expressing M2 macrophages are elevated in the peritoneal fluid of patients with metastasis of gastric cancer

Background: A critical immunoregulatory pathway in the tumor microenvironment (TME) involves CD39 and related cell-surface located enzymes that regulate the hydrolysis of extracellular, pro-inflammatory ATP and generate immunoregulatory adenosine. In gastric cancer, CD39 is upregulated and associated with worse survival, but little is known about the role of CD39 in peritoneal metastasis. In this study, we investigate immune cell expressed CD39 in peritoneal fluid as part of the TME of peritoneal metastasis derived from gastric cancer.

Immune cell-derived microparticles in blood and peritoneal fluid of patients with peritoneal metastasis of gastric cancer

Background: Peritoneal metastasis (PM) of gastric cancer (GC) origin is still associated with poor survival. Microparticles, small lipid-bilayer coated extracellular vesicles shed in the tumor microenvironment by immune and tumor cells, enhance tumor growth and metastatic potential of tumor cells by various mechanisms including the induction of an immunoregulatory microenvironment via degradation of ATP to its derivates ADP, AMP and adenosine by membrane-bound ectonucleotidases such as CD39. In this study, we explore the role of immune cell derived microparticles in GC derived PM.