Peritoneal Fluid Samples Research Articles

Extracellular Vesicle Characteristics in Local Fluid and Plasma Measured by Nanoparticle Tracking Analysis Can Help Differentiate High-Grade Serous Carcinoma from Benign Ovarian Pathology

Background: High-grade serous carcinoma (HGSC) is the most lethal of gynecological cancers in developed countries. It usually presents late with non-specific symptoms and most cases are diagnosed at an advanced stage, with 5-year overall survival being around 40%. Biomarkers for screening and early diagnosis of this aggressive disease are, thus, a research priority. Extracellular vesicles (EVs) that reflect the cell of origin and that can be isolated from local fluid and plasma by minimally invasive liquid biopsy are such promising biomarkers. Besides EV concentration and molecular profile, which have been the main focus of research for many years, recent studies have also called attention to EV size distribution. The aim of our study was to evaluate the potential of EV concentration and size distribution in local fluid and plasma as diagnostic biomarkers for HGSC. Methods: Paired pretreatment ascites and plasma samples from 37 patients with advanced HGSC and paired pretreatment free peritoneal fluid (FPF) and plasma samples from 40 controls with benign ovarian pathology (BOP) were analyzed using nanoparticle tracking analysis (NTA). Results: We observed a significant difference in EV concentration in local fluid, but not in plasma, between HGSC patients and the control group. We also found a significant difference in EV size distribution in both local fluid and plasma between HGSC patients and the control group. The receiver operating characteristics (ROC) curve analysis of EV characteristics showed excellent diagnostic performance for the mode, D10, and D50 in local fluid and acceptable diagnostic performance for EV concentration and mean EV size in local fluid, as well as for the mode and D10 value in plasma. Conclusions: The results of our study show that EV concentration in local fluid and more importantly EV size distribution in both local fluid and plasma are significantly changed in the presence of HGSC. Future research of size-dependent molecular profiling of EVs could help identify novel diagnostic biomarkers for HGSC.

The Role of NK and T Cells in Endometriosis.

Endometriosis, a debilitating condition, affects one in ten women of reproductive age. Its pathophysiology remains unclear, though deficiencies in immune surveillance are thought to create an environment conducive to the evasion of ectopic endometrial cells from the immune system. Our research explores the immunological impact of endometriosis both locally and systemically, emphasizing natural killer (NK) and T cell subpopulations. We incorporated 62 female patients who underwent laparoscopic surgery; of those, 47 had endometriosis, and 15 were controls. We collected peritoneal fluid (PF) and peripheral blood (PB) samples which were tagged with monoclonal antibodies and subsequently scrutinized using flow cytometry. Our findings revealed significant differences in immunological profiles based on demographic factors and symptomatology. In the endometriosis cohort, there was an increase in PB CD56HiCD16dim and PF CD8+ CD56dimCD16Hi NK cells. CD16+ CD4 T cell levels were significantly lower in the PB of endometriosis patients who smoke. Individuals with more severe disease displayed significantly higher levels of PB CD16+ CD8 T cells, which also increased in those with non-menstrual pelvic pain. Dysmenorrhea severity correlated with a progressive increase in PF CD8+ CD56dimCD16Hi NK cells. These variations in specific lymphocyte subsets, namely, within NK and T cells, suggest potential immunological mechanisms in the evolution and clinical presentation of endometriosis.

A Challenging Diagnosis - Large Leiomyomas on Rudimentary Uterus in Mayer-Rokitansky-Küster-Hauser (MRKH) Syndrome

A 40-year-old woman diagnosed with Mayer-Rokitansky-Küster-Hauser (MRKH) syndrome at 16 years of age presented with a large abdominal mass protruding to the right subcostal margin, equivalent to 30 weeks gestation. She didn’t have comorbidities of hypertension or diabetes. The vitals were normal with BMI 30. She was asymptomatic except for occasional vague abdominal discomfort. Further imaging studies were conducted to assess the extent and nature of the findings. Pelvic and transvaginal ultrasound demonstrated the presence of multiple large fibroids extending beyond the level of the umbilicus. This was corroborated by pelvic MRI, which further confirmed the massively enlarged multi-fibroid uterus. Additionally, intravenous contrast-enhanced imaging revealed large bilateral ovarian collaterals and prominent blood vessels extending to the broad ligament. Triple-phase CT angiogram of the abdomen/pelvis with pre-contrast revealed an 8mm aneurysm on the anterior aspect of the abdominal aorta and unusual vascular anatomy, raising the possibility of an atypical or neoplastic process in the right iliac fossa. In view of these findings, an opinion by an Oncologist was sought, who suspected the possible presence of an abnormal nidus of vessels and a fluid attenuation area in the right iliac fossa. Additionally, the Risk of Ovarian Malignancy Algorithm (ROMA) test was performed, revealing a slightly elevated value. To reach the final diagnosis, the decision for the exploratory laparotomy was undertaken. Intraoperative findings revealed the presence of a rudimentary uterus with normal ovaries (Figure 1 and Figure 2), as well as the presence of bilateral broad fibroids (Figure 3). The broad ligament areas around the uterus were occupied with large circumscribed masses, identified as fibroids. The right-side fibroid measuring 17 x 15cm (Figure 4) and the left-side fibroid measuring 10 x 10cm (Figure 5) were excised, respectively, while preserving the left ovary and rudimentary uterus. Peritoneal fluid samples were obtained to ascertain the nature of malignancy. No evidence of infiltration or adhesions was detected. Postoperatively, the patient experienced no complications. Histological examination of the masses growing from bilateral uterine remnants confirmed the presence of leiomyomas, with no glandular epithelium identified. Peritoneal fluid analysis revealed the absence of malignant cells.

Peritoneal-dialysis-associated peritonitis due to Staphylococcus sciuri: a case report and literature review

BackgroundPeritoneal-dialysis-associated peritonitis is a serious complication of peritoneal dialysis; prevention and treatment are crucial to reduce patient morbidity and mortality. Staphylococcus sciuri is an organism that, although present in several animal species, rarely causes disease in humans, particularly in patients undergoing peritoneal dialysis.Case presentationA 54-year-old man with end-stage kidney disease on continuous ambulatory peritoneal dialysis was admitted to the emergency department with nausea, vomiting, slight abdominal distention, and cloudy peritoneal dialysis effluent. Relevant medical history included occupational exposure to different animal species. The white blood cell count in the peritoneal dialysis fluid sample was 11,200 cells/mm3 with 90% polymorphonuclear cells. Peritoneal-dialysis-associated peritonitis was diagnosed. Empirical antibiotic treatment was started according to the International Society for Peritoneal Dialysis guidelines. Staphylococcus sciuri was identified in the peritoneal dialysis effluent culture by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The treatment was modified on the basis of the sensitivity results. Medical cure was established on the basis of the negative result of the dialysate culture and absence of alterations in the cell count with differential after the end of treatment. The patient was instructed to be careful when administering peritoneal dialysis, especially in the care involved in being in contact with animals.ConclusionsThis article highlights the importance of Staphylococcus sciuri as a significant pathogen due to its virulence and antibiotic resistance, especially in patients undergoing peritoneal dialysis. Prevention strategies as well as timely diagnosis and treatment compliance according to the International Society for Peritoneal Dialysis are essential for successful outcomes.

O-267 Extracellular vesicles as biomarkers of endometriosis

Abstract Study question Can extracellular vesicles serve as biomarkers of endometriosis, particularly for stages I and II, which are difficult to detect by ultrasound? Summary answer Our findings suggest the feasibility of defining a protein signature of endometriosis based on EV cargoes. What is known already Endometriosis is characterised by intraperitoneal lesions of endometrial tissue. Although ultrasound detection of severe endometriosis is possible, the diagnosis of peritoneal lesions currently relies on invasive laparoscopy, which leads to a delay of 8-10 years. We have previously shown that peritoneal fluid (PF) of women with endometriosis contains endometriosis specific EVs. Traced in peripheral blood (PB), EVs could serve as biomarkers of endometriosis, particularly for stages I and II, which are difficult to detect by ultrasound. Study design, size, duration Women 18-45 years of age undergoing laparoscopic surgery for endometriosis or unrelated conditions were invited to participate in our study (HREC 20-159A/HREC 20-882A). Following informed consent, we collected PF and PB samples from n = 101 participants (n = 47 controls, n = 54 cases). Exclusion criteria were pregnancy, malignancy, and menopause. Participants/materials, setting, methods We processed PF and PB samples by differential ultracentrifugation and validated the presence of EVs by nanoparticle tracking analysis (NTA), Western blotting and transmission electron microscopy (TEM). EVs were analysed by untargeted, label-based quantitative proteomics using Tandem Mass Tags (TMT). Data analysis was performed using Proteome Discoverer v2.4 (Thermo Fisher) followed by statistical analysis using R. Candidate biomarker proteins were tested by traditional and automated Western blotting (WES, ProteinSimple). Main results and the role of chance We ascertained the identity of EVs by TEM and NTA (mode size 121.8 ± 18.0 nm (blood, n = 5) and 155.9 ± 37.2 (PF, n = 6)), and signals for syntenin and ALIX in immunoblots as well as expression of CD9, CD63 and CD81 in capture bead-flow cytometry. Proteomic analysis identified 9145 protein groups across all sample groups, with 602 significantly regulated between comparisons (adjusted P-value <0.05). In PF, 533 proteins changed significantly in abundance (245 up/288 down), while in blood, only four proteins changed in abundance. We tested the presence/absence of two of these as candidate markers on PB EV protein preparations from n = 32 samples to date; our best marker showed a sensitivity of 67% and a specificity of 90%; upon exclusion of severe endometriosis cases, sensitivity improved to 80%. Limitations, reasons for caution Because of the changes in vesicle composition and amounts with the menstrual cycle, we have to group the samples by menstrual cycle phase and disease severity. This leaves relatively small numbers per group per condition. Another limitation concerns the relatively small amount of sample liquid and thus, EVs available for analysis. Wider implications of the findings Our findings suggest the feasibility of defining a protein signature of endometriosis based on EV cargoes. A non-invasive clinical test would allow for earlier diagnosis and thus treatment of endometriosis. It would also allow patients to initiate fertility treatment early. Trial registration number not applicable

Microbiological Profiles of the Anatomical Sites of Perforation Peritonitis: A Cross-Sectional Study.

Introduction One of the most frequent emergencies that a general surgeon deals with is perforation peritonitis. The anatomical site of the perforation, which in turn affects the source of infection, has a major impact on the mortality rate due to perforation peritonitis. Early and suitable antibiotic therapy can be started in the postoperative period with the aid of knowledge about the microbiological profile and sensitivity of peritoneal fluid culture with respect to the anatomical sites of perforation peritonitis. Methods A cross-sectional study was conducted from June 2021 to November 2021 where peritoneal fluid samples were collected intraoperatively from patients with perforation peritonitis. This was subjected to culture and sensitivity, and results were analyzed with respect to anatomical sites of perforation. Results Forty cases were investigated. The ileum (30%) was the most common site of perforation, followed by the stomach (22.5%), appendix (20%), duodenum (12.5%), caecum (5%), jejunum (5%), transverse colon (2.5%), and rectum (2.5%). Escherichia coli (E. coli) and Klebsiella spp. were the most frequently found organisms in all sites of perforation peritonitis. The most sensitive antibiotics covering all isolated organisms were amikacin and meropenem. Sensitivity to amikacin was found in 85.18% of cases of E. coli and 84.6% of cases of Klebsiella. Sensitivity to meropenem was found in 76.9% of cases of E. coli and 80% of cases of Klebsiella. Conclusion In patients with perforation peritonitis, the peritoneal fluid cultures did not reflect the major differential normal flora according to the region of the gastrointestinal tract. The most prevalent organism isolated among all the sites of perforation peritonitis was E. coli. Antimicrobial activity against organisms isolated from perforation peritonitis patients was significantly demonstrated by aminoglycosides, piperacillin and tazobactam, and meropenem and colistin, with considerable resistance to third-generation cephalosporins.

Oral Treatment with the Pectin Fibre Obtained from Yellow Passion Fruit Peels Worsens Sepsis Outcome in Mice by Affecting the Intestinal Barrier.

The biological activities of plant-derived soluble dietary fibres (SDFs) have been widely investigated. Pectin from yellow passion fruit (YPF-peSDF) peels was suggested as a protective macromolecule in ulcers and colitis due to its antioxidant and anti-inflammatory properties. Sepsis has high mortality and morbidity and is characterised by inflammatory and oxidative stress imbalances. Evidence suggests that pectins may aid sepsis treatment; however, the effects of YPF-peSDF on sepsis remain unclear. Herein, polymicrobial sepsis was induced by cecal-ligation and puncture in mice treated with YPF-peSDF (1 and 10 mg/kg; gavage). YPF-peSDF accelerated mortality, reaching 100% in 24 h. Inflammation was present in the colons and small intestines (SI) of both vehicle- and fibre-treated mice. Although crypt depth and width, and villus height were preserved in the SI of septic mice administered YPF-peSDF, they exhibited exacerbated muscle layer atrophy and mucosa and submucosa hypertrophy, along with shortened enterocytes. Larger crypts and shorter enterocytes were noted in their colons in comparison with vehicle-controls. YPF-peSDF also reduced inflammatory cell numbers and exacerbated IL-6 levels in peritoneal lavage fluid (PELF) samples. YPF-peSDF modulated SI but not colon cytokines. Lipoperoxidation and antioxidant capacity levels were attenuated in PELF samples. Overall, in contrast to previous evidence, YPF-peSDF worsened polymicrobial sepsis outcomes in mice.

TIM-3 Expression in Endometriosis.

Endometriosis is a prevalent chronic gynecological disease linked to immune dysfunction. The protein T-cell immunoglobulin and mucin domain-containing protein 3 (TIM-3) plays a crucial role in immune system balance. Malfunction of TIM-3 may result in excessive immune activation and inflammatory tissue damage. Given TIM-3's established role in the development of cancer and autoimmune diseases, we decided to study its role in women suffering from endometriosis. We included a total of 62 female patients, all of whom had undergone laparoscopic surgery. Of these, 47 had endometriosis and 15 did not. During surgery, we collected peritoneal fluid (PF) and peripheral blood (PB) samples from every patient for analysis using flow cytometry. To mark the samples, we used a panel of monoclonal antibodies and examined TIM-3 expression in their immune cells. Endometriosis patients in PB demonstrated a significantly lower percentage of CD56+ T cells with TIM-3 expression. As endometriosis progressed through its stages, this expression lessened. This decrease was particularly notable in women with stage III/IV endometriosis. Additionally, both women diagnosed with intestinal endometriosis and those with recent endometriosis diagnoses showed a significantly reduced percentage of CD56+ T cells expressing TIM-3. Patients with endometriosis exhibit diminished TIM-3 expression within circulating T cells. This warrants further investigation to discern whether it contributes to the progression of endometriosis, potentially through the amplification of autoreactive T cells and inflammation.

Intraperitoneal pharmacokinetics of systemic oxaliplatin, 5-fluorouracil and bevacizumab in patients with colorectal peritoneal metastases

BackgroundPeritoneal metastases (PM) commonly occur in colorectal cancer patients. Systemic chemotherapy yields poor outcomes for these patients. It is hypothesised that traditional systemic chemotherapy is not very effective for this patient population. This study investigates to what extent systemic anti-cancer therapy crosses the peritoneal barrier. MethodsIn a Phase I study, eighteen patients received systemic oxaliplatin, 5-FU, and bevacizumab. Plasma and peritoneal fluid samples were collected to measure drug concentrations. A non-compartmental analysis determined the Area Under the Curve (AUC) for oxaliplatin and 5-FU in both matrices. Intraperitoneal (IP) and intravenous (IV) exposure ratios were calculated, along with the bevacizumab concentration IP/IV ratio. The relationship between tumour load and IP/IV ratios and the correlation between the IP/IV ratios of different treatments were assessed statistically. ResultsA total of 438 5-FU samples and 578 oxaliplatin samples were analysed in plasma and peritoneal fluid. Bevacizumab was quantified with 17 measurements in plasma and 15 measurements IP. Median IP/IV ratios were 0.143, 0.352 and 0.085 for 5-FU, oxaliplatin and bevacizumab, respectively. Oxaliplatin exhibited a longer IP half-life than 5-FU. A correlation was found between oxaliplatin and bevacizumab IP/IV ratios (R=0.69, p=0.01). No statistical correlations were found between the other investigated drugs. ConclusionsOur findings indicate that only a small percentage of systemically administered anti-cancer treatment reaches the IP cavity, questioning their efficacy against PM. This strengthens the hypothesis for repeated intraperitoneal chemotherapy to reach adequate anti-cancer drug levels.

Exosomal ROR1 in peritoneal fluid identifies peritoneal disseminated PDAC and is associated with poor survival.

Pancreatic ductal adenocarcinoma (PDAC) is one of the deadliest forms of cancer and peritoneal dissemination is one major cause for this poor prognosis. Exosomes have emerged as promising biomarkers for gastrointestinal cancers and can be found in all kinds of bodily fluids, also in peritoneal fluid (PF). This is a unique sample due to its closeness to gastrointestinal malignancies. The receptor tyrosine kinase-like orphan receptor 1 (ROR1) has been identified as a potential biomarker in human cancers and represents a promising target for an immunotherapy approach, which could be considered for future treatment strategies. Here we prospectively analyzed the exosomal surface protein ROR1 (exo-ROR1) in PF in localized PDAC patients (PER-) on the one hand and peritoneal disseminated tumor stages (PER+) on the other hand followed by the correlation of exo-ROR1 with clinical-pathological parameters. Exosomes were isolated from PF and plasma samples of non-cancerous (NC) (n = 15), chronic pancreatitis (CP) (n = 4), localized PDAC (PER-) (n = 18) and peritoneal disseminated PDAC (PER+) (n = 9) patients and the surface protein ROR1 was detected via FACS analysis. Additionally, soluble ROR1 in PF was analyzed. ROR1 expression in tissue was investigated using western blots (WB), qPCR, and immunohistochemistry (IHC). Exosome isolation was proven by Nano Tracking Analysis (NTA), WB, Transmission electron microscopy (TEM), and BCA protein assay. The results were correlated with clinical data and survival analysis was performed. PDAC (PER+) patients have the highest exo-ROR1 values in PF and can be discriminated from NC (p <0.0001), PDAC (PER-) (p <0.0001), and CP (p = 0.0112). PDAC (PER-) can be discriminated from NC (p = 0.0003). In plasma, exo-ROR1 is not able to distinguish between the groups. While there is no expression of ROR1 in the exocrine pancreatic tissue, PDAC and peritoneal metastasis show expression of ROR1. High exo-ROR1 expression in PF is associated with lower overall survival (p = 0.0482). With exo-ROR1 in PF we found a promising diagnostic and prognostic biomarker possibly discriminating between NC, PDAC (PER-) and PDAC (PER+) and might shed light on future diagnostic and therapeutic concepts in PDAC.

Pet-Related Pasteurella Multocida-Induced Peritonitis in Peritoneal Dialysis: A Case Report

Pasteurella multocida is a small, Gram-negative cocco-bacillus usually found in the oral cavities of most healthy cats and dogs as part of their natural oral flora. This zoonotic pathogen can cause a variety of infections in humans. The number of reported cases of peritoneal dialysis-associated peritonitis caused by P. multocida has been limited worldwide. We report here the case of a 38-year-old woman on continuous peritoneal dialysis (PD) that presented to the emergency department with abdominal pain and vomiting lasting for two days. A sample of peritoneal dialysis fluid was explored in the microbiology laboratory where grown microorganisms was identified by the VITEK2 system as P. canis with 90% accuracy. Then whole genome sequencing concluded that the species is rather P. multocida and that the strain was sensitive to all antibiotics tested and harbored various virulence genes. Patient was treated by intraperitoneal antibiotic course with cefotaxime for 3 weeks.

Oxidative stress and antioxidant capacity in patients with endometrioma.

BackgroundEndometriosis has several clinical features, including dysmenorrhea, infertility, and endometrioma (EMO). Although oxidative stress status is closely related to endometriosis, it is unclear how the balance between oxidative stress capacity and antioxidant capacity correlates with treatment of or factors that worsen endometriosis. In this study, we used peritoneal fluid from patients with EMO to investigate the role of oxidative stress capacity and antioxidant capacity.Materials and MethodsParticipants with EMO (n = 30) and without EMO (uterine myoma, n = 13) were enrolled. All peritoneal fluid samples were collected at the beginning of surgery. We evaluated oxidative stress capacity and antioxidant capacity in peritoneal fluid samples by using the diacron-reactive oxygen metabolites (d-ROM) and biological antioxidant potential (BAP) tests, respectively. The d-ROM and BAP values and the d-ROM/BAP ratio were measured, and their correlations with the CA125 level, revised American Society for Reproductive Medicine (r-ASRM) score, and tumor size were analyzed.ResultsThe d-ROM/BAP ratio was significantly higher in patients with EMO than in those without EMO. In addition, the d-ROM/BAP ratio was positively correlated with CA125 level and r-ASRM scores in patients with EMO.ConclusionsOxidative stress is correlated with factors that worsen EMO. The d-ROM/BAP test may be useful for assessing disease status in patients with EMO.

The SPRi determination of cathepsin L and S in plasma and peritoneal fluid of women with endometriosis

PurposeEndometriosis is a common disease with a complex pathomechanism and atypical symptoms, often leading to delayed diagnosis. Currently, the sole method for confirming the presence of the disease is through laparoscopy and histopathological examination of collected tissue. However, this invasive procedure carries potential risk and complications, necessitating the exploration of non-surgical diagnostic methods for endometriosis. This study aims to analyze peritoneal fluid and plasma samples for the expression of cathepsin L and cathepsin S to identify potential biomarkers for non-invasive diagnostic approaches to endometriosis. Material and methodsIn this cross-sectional study, plasma and peritoneal fluid samples were obtained during laparoscopy from 63 patients diagnosed with chronic pelvic pain or infertility. The study group consisted of women with confirmed endometriosis. The concentrations of cathepsins L and S were determined using an SPRi biosensor. ResultsThe study did not reveal significant differences in the concentrations of cathepsin L and cathepsin S between the control group and the study group, both in peritoneal fluid and plasma. ConclusionsBased on the results of this study, it appears that cathepsins L and S are not suitable candidates as biomarkers for endometriosis.

Listeria monocytogenes-associated spontaneous bacterial peritonitis in France: a nationwide observational study of 208 cases

Listeriosis is a foodborne infection caused by Listeria monocytogenes. Three main forms of listeriosis are well characterised, but little is known about L monocytogenes-associated spontaneous bacterial peritonitis. We used data from the French national surveillance of listeriosis to perform a nationwide retrospective study. All patients with L monocytogenes isolated by culture from a peritoneal fluid sample in France between April 1, 1993, and Dec 31, 2022, were included. Individuals for whom bacterial peritonitis was not confirmed and those who also had another type of invasive listeriosis were excluded. A standardised checklist was used to collect demographic, clinical, and biological data as well as antibiotic treatment and follow-up data. The primary outcome was to determine the characteristics of L monocytogenes-associated spontaneous bacterial peritonitis. We did descriptive analyses and assessed risk factors for 1-month mortality using an exploratory multivariable Cox model analysis. Among the 8768 L monocytogenes cases reported, 208 (2%) were patients with L monocytogenes-associated spontaneous bacterial peritonitis. Mean age was 65 years (SD 13), 50 (24%) of 208 patients were female, and 158 (76%) were male (no data on race or ethnicity were available). 200 (98%) of 205 patients with L monocytogenes-associated spontaneous bacterial peritonitis with available data had immunosuppressive comorbidities, including cirrhosis (148 [74%] of 201 with available data), ongoing alcoholism (58 [62%] of 94), and ongoing neoplasia (60 [31%] of 195). Causes of ascites included cirrhosis (146 [70%] of 208), ongoing neoplasia (26 [13%]), end-stage heart failure (13 [6%]), and peritoneal dialysis (11 [5%]). Among those with available data, presentation was pauci-symptomatic and non-specific; only 67 (50%) of 135 patients presented with fever, 49 (37%) of 132 with abdominal pain, and 27 (21%) of 129 with diarrhoea. 61 (29%) of 208 patients were dead at 1 month, 92 (44%) were dead at 3 months, and 109 (52%) were dead at 6 months after diagnosis. Ongoing neoplasia (hazard ratio 2·42 [95% CI 1·05-5·56]; p=0·039), septic shock (8·03 [2·66-24·02]; p=0·0021), and high blood leukocyte count (1·05 [1·00-1·09]; p=0·045) were independently associated with 1-month mortality. Despite the non-specific and mild presentation of L monocytogenes-associated spontaneous bacterial peritonitis, the outcome is poor and similar to that of neurolisteriosis, and so identification of L monocytogenes in ascitic fluid samples requires urgent parenteral amoxicillin-based treatment to avoid a fatal outcome. Institut Pasteur, Inserm, and French Public Health Agency. For the French translation of the abstract see Supplementary Materials section.

An Uncommon Cause of Appendicitis “Actinomyces Turicensis,” A Rare Species Causing Abdominal Actinomycosis

Abstract Primary actinomycosis of the appendix is a rare clinical entity. It is generally diagnosed based on histological examination or culture of actinomyces obtained from a peritoneal fluid sampling, and the treatment frequently requires surgical resection.

Role of TNF superfamily members lymphotoxin-α, sCD40L, and TNF-α in endometriosis-related infertility

Introduction: TNF-α, LT-α, and sCD40L belong to the TNF superfamily and play a pivotal role in chronic inflammatory disease. Yet, these soluble molecules are largely unexplored in endometriosis cases with infertility. The present study aimed to evaluate serum and peritoneal fluid levels of these molecules and correlate its level to the endometriosis severity. Methods: Peripheral blood and peritoneal fluid samples were obtained from 87 infertility cases who underwent laparoscopy, consisting of 44 endometriotic women and 43 non-endometriotic women. The levels of TNF-α, LT-α, and sCD40L were determined using ELISA. Results: Serum and peritoneal TNF-α levels were significantly elevated in the endometriosis group compared to the control group ( p = 0.028; p = 0.027), but not the sCD40L and LT-α level. Serum and peritoneal TNF-α levels were significantly different between late-endometriosis (stage III or IV) compared to the control group ( p = 0.026, p = 0.041, respectively). Moreover, the serum LT-α level was significantly higher in late-endometriosis cases compared to the early-endometriosis (stage I or II) cases ( p = 0.03) and showed a positive correlation with endometriosis rASRM score ( p = 0.001). ROC curve analysis showed a significant diagnostic value of serum LT-α in differentiating the late cases to the early endometriosis patients (AUC = 0.751, p = 0.007). A negative correlation between LT-α and total Endometriosis Fertility Index (EFI) score was also observed ( p = 0.012). Conclusion: Severe endometriosis and lower EFI score are associated with higher serum LT-α level. Serum LT-α is a potential staging biomarker among endometriosis cases. Serum and peritoneal TNF-α levels were elevated in women with severe endometriosis among infertility cases. Further study should address the role and predictive value of LT-α in endometriosis-related infertility.

Diagnostic utility of the total nucleated cell count for differentiation of septic and sterile peritoneal effusions in dogs.

Rapid and accurate diagnosis of septic peritonitis is critical for initiating appropriate medical and surgical management. The aim of this study was to determine the diagnostic utility of the total nucleated cell count (TNCC), absolute neutrophil count, neutrophil percentage, and total protein (TP) to distinguish septic versus non-septic peritoneal effusions in dogs. Electronic medical records were retrospectively searched for peritoneal fluid samples from 2008 to 2018 and classified as septic or non-septic based on bacterial culture and/or cytology results. Receiver operator characteristic curves (ROCs) were used to describe the overall diagnostic utility of each test, with optimal cutpoints analyzed to dichotomize continuous variables. Positive and negative likelihood ratios were calculated at these cutpoints. A total of 166 unique samples, including 87 septic and 79 non-septic peritoneal effusions, were included. There were no significant differences in dog sex, age, or days hospitalized between groups. Septic effusions had significantly higher TP, TNCC, absolute neutrophil count, and neutrophil percentage compared with non-septic effusions. The area under the curve of the ROC curves was TNCC (0.80), absolute neutrophil count (0.80), neutrophil percentage (0.64), and TP (0.63). For TNCC and absolute neutrophil count, optimal cutoffs were 17.13 × 103 cells/μL and 19.88 × 103 cells/μL, resulting in positive and negative likelihood ratios of 2.39 and 0.28 and 2.85 and 0.28, respectively. Total nucleated cell counts and absolute neutrophil counts aid in the differentiation of septic and non-septic peritoneal effusions with similar diagnostic utility but are not sufficiently sensitive or specific to use without concurrent microscopic evaluation.

The Use of the Neoglycolipid-Based Oligosaccharide Microarray System in the Diagnosis of Endometriosis - Preliminary Study.

Endometriosis presents diagnostic challenges, and there is a need for developing novel biomarkers with satisfactory specificity and sensitivity. Glycomics, exploring glycosylation changes in glycoproteins, offers potential solutions. The aim of this study was to analyze the carbohydrate-binding properties of IgG and IgM antibodies in the plasma and peritoneal fluid samples and to identify any differences in the presence and the specificities of anti-carbohydrate antibodies in the endometriosis patient and the controls. Multicenter study was conducted in Poland between 2018 and 2019. Plasma and peritoneal fluid samples were collected from women undergoing laparoscopic surgery. Endometriosis patients (n=8) and controls (n=8), matched for cycle phase and disease stage, were selected. The neoglycolipid-based oligosaccharide microarray system was used to investigate IgG and IgM antibody binding properties to glycan-related probes in biological materials. In peritoneal fluid samples, IgM binding to the following probes was significantly higher in endometriosis: GSC-915-4 (new), LNFP-I, NeuAcα-(6')LNnO (F1), B-like decaosylceramide, log10(GM1-penta), and log10(GSC-915-5). In a control group higher IgG binding to log10(Orsay-5-AO) was observed. In plasma samples, endometriosis showed higher IgG binding to log10(NeuAcα-(6')LNnO (F1)) and lower IgG binding to Gal2GlcNAc(1-3)-AO. After Benjamin-Hochberg correction, differences were not significant. Effect sizes highlighted some glycan probes in both plasma and peritoneal fluid. Strong correlations were observed among binding to certain glycan probes. This preliminary study suggests glycomics' potential contribution to endometriosis diagnosis and understanding of its pathophysiology. Neoglycolipid-based microarrays hold promise for non-invasive endometriosis diagnostic tools. Further investigations with larger cohorts are warranted to validate these findings and explore potential correlations with antibody levels in plasma and peritoneal fluid. Glycomics emerges as a valuable diagnostic asset in endometriosis research.

Clinical characteristics and bacterial culture in complicated appendicitis at Hue University of Medicine and Pharmacy Hospital

Objectives: To evaluate the clinical, laboratory features, bacterial culture outcomes, and antibiotic susceptibilities of these bacteria in intraoperative samples from patients with perforated appendicitis and to establish whether they influence postoperative outcomes. Materials and methods: A descriptive study was conducted on 68 laparoscopic surgery patients from June 2022 to March 2023 at the Digestive Surgery Department, Hue University of Medicine and Pharmacy Hospital. Results: The male/female ratio is 1.2. The mean age was 37.5 ± 25.7. The mean time of the symptom onset was 37.2 ± 29.1 hours; CRP increased in 86.8% of cases. The average postoperative hospital stay was 6.3 ± 2.2 days; Postoperative complications were observed in 20.6%. Peritoneal fluid samples yielded a positive culture in 58 (85.3%); Gram-negative bacteria were isolated in 89.7%; gram-positive bacteria were isolated in 10.3%; polymicrobial growth was observed in 29.4% of the patients. E. coli in 67.6%, P.aeruginosa in 10.3%, Enterobacter spp. in 8.8%, Klebsiella spp. in 4.4%, Citrobacter spp. in 3%; positive ESBL was present in 17 (37%) of the 46 E. coli strains, ESBL was positivity detected in one of Morganerlla morganii strains. In E. coli, ampicillin resistance was 88.6%, amoxicillin-clavulanic acid resistance was 61.5%, and Trimethoprim-sulfamethoxazole resistance was 65%. Conclusion: It is necessary to choose treatment regimens based on local bacteriology and resistance status to optimize empiric empirical postoperative antibiotic regimens. Key words: perforated appendicitis, ESBL, complications, bacteria culture.

Diagnóstico clínico e anatomopatológico em bovinos acometidos por úlceras do abomaso perfuradas e com comorbidades

This study describes the findings of the clinical and anatomopathological diagnosis in cattle affected by perforated abomasal ulcers and with comorbidities. Twenty-five cattle were evaluated, young (n=8, aged less than two years) and adults (n=17, aged greater than two and a half years), diagnosed with perforated abomasal ulcers. Epidemiological information was retrieved from the medical records. Blood, ruminal fluid, and peritoneal fluid (PI) samples were collected for laboratory examination. Ultrasound examination, exploratory laparotomy, and post mortem examinations were performed for diagnostic purposes. The main clinical findings of perforated ulcers in the present study were: apathy, dehydration, intestinal hypomotility, diarrheal stools, hypomotile or atonic rumen, tachycardia, tachypnea, abdominal distention with bilateral bulging, and increased abdominal tension. The most frequent abnormal hematological findings were hypoproteinemia and neutrophilic leukocytosis with a left shift. The peritoneal fluid (PL) showed an inflammatory reaction in the cytology, characteristic of an exudate. A confirmatory diagnosis of peritonitis by ultrasonography was possible in five cases, and in two cases at exploratory laparotomy. In the post mortem examination, the ulcers were characterized as type 3, 4, or 5, with comorbidities present in 68% of the cases. It is necessary to combine ultrasonography with abdominal centesis and exploratory laparotomy procedures to suggest perforated ulcers. However, post mortem examination better characterizes the type of perforated ulcer present, particularly type 5.