Fluid Samples Research Articles

No significant difference in skin contamination during anterior cruciate ligament reconstruction with and without preoperative skin cleaning.

This prospective study aimed to assess whether preoperative antiseptic skin cleansing reduces bacterial contamination and surgical site infections (SSI) following anterior cruciate ligament reconstruction (ACLR). We hypothesized that antiseptic cleaning would lower bacterial load, reducing contamination and early infections. One hundred and nineteen patients scheduled for ACLR were included in this prospective, nonrandomized study. Individuals were divided into two groups. Patients in the intervention group applied octenisan® wash lotion daily for three days before surgery and used the wash solution instead of their usual shower gel. Additionally, they swiped their leg with octenisan® soaked gloves on the morning of the operation. The control group followed their usual wash routine with no specific instructions. Fluid samples were taken before surgery from the irrigation bag and at 15-min intervals from the reservoir of the sterile surgical drape during the procedure. Suture material used for the ACL graft and meniscus repair were also collected for testing. The samples were subjected to a 14-day incubation period. Follow-up included outpatient visits at 6 weeks, 12 weeks and 6 months with a final evaluation at 12 months. Contamination rates showed no significant difference between the control and intervention groups. The mean contamination rate in the control group was 6.4% (n = 22) and 6.6% (n = 24) in the intervention group (p = 0.28). At 12-month follow-up, 110 out of 119 participants were included (52 control, 58 intervention). T tests for age (p = 0.19), BMI (p = 0.66), and surgery duration (p = 0.38) showed no significant differences. No early SSI were observed in either group postoperatively. Our results indicate that the use of antiseptic wash lotion and gloves does not influence the risk of bacterial contamination during surgery. Level III.

Liquid biopsy for CNS lymphoma: CSF exosomes and CSF exosomal miR-15a, miR-21, miR-155, miR-210, and miR-19b are promising biomarkers for diagnosis.

Central nervous system lymphoma (CNSL) is a devastating disease with a poor prognosis. Early diagnosis, monitoring of the treatment response, and outcome prediction carry the utmost importance in the management of patients with CNSL. Surgical biopsy is the gold standard for tissue diagnosis, however, this procedure has potential complications. Therefore, there is a need for a method that provides information about diagnosis and patient monitoring to avoid surgical risks. The study aimed to investigate potential diagnostic biomarkers for patients with CNSL. Patients with secondary CNSL were included in this study. Serum and cerebrospinal fluid (CSF) samples were collected before treatment and after completion of the treatment. Cell-free DNA (cfDNA), exosomes, free and exosomal microRNA (miR)-15a, miR-21, miR-155, miR-210, and miR-19b in both serum and CSF were examined, and they were compared with the controls. Also, their levels before and after treatment were compared. Nine patients with the diagnosis of secondary CNSL were reviewed. cfDNA, miR-15a, and miR-155 in serum, and exosome in CSF were found to be significantly higher in CNSL patients compared to the controls. Exosomal miR-15a, miR-21, miR-155, miR-210, and miR-19b in CSF were found to be significantly higher in CNSL patients compared to controls, whereas their levels in serum were not significantly high. Our findings suggested that exosomes and exosomal miR-15a, miR-21, miR-155, miR-210 and miR-19b in CSF would be promising biomarkers for the diagnosis of patients with CNSL. Further studies are needed to confirm our findings.

Augmented pathogen detection in brain abscess using metagenomic next-generation sequencing: a retrospective cohort study.

Brain abscess is a severe infection characterized by the accumulation of pus within the brain parenchyma. Accurate identification of the causative pathogens is crucial for effective treatment and improved patient outcomes. This 10-year retrospective, single-center study aimed to compare the detection performance of conventional culture methods and metagenomic next-generation sequencing (mNGS) in brain abscess. We reviewed 612 patients diagnosed with brain abscess and identified 174 cases with confirmed etiology. The median age was 52 years, with 69.5% males. Culture tests predominately identified gram-positive bacteria, particularly Streptococcus spp. Gram-negative bacteria, including Klebsiella spp., were also detected. However, mNGS revealed a more diverse pathogen spectrum, focusing on anaerobes (e.g., Fusobacterium spp., Parvimonas spp., Porphyromonas spp., Prevotella spp., and Tannerella spp.). mNGS exhibited significantly higher overall pathogen-positive rates in pus samples (85.0% vs 50.0%, P = 0.0181) and CSF samples (84.2% vs 7.9%, P < 0.0001) compared to culture. Furthermore, the detection rates for anaerobes displayed a notable disparity, with mNGS yielding significantly higher positive detections in both pus samples (50.0% vs 10%, P = 0.0058) and CSF samples (18.4% vs 0%, P = 0.0115) when compared to culture methods. The assistance of mNGS in pathogen detection, particularly anaerobes in brain abscess, was evident in our findings. mNGS demonstrated the ability to identify rare and fastidious pathogens, even in culture-negative cases. These results emphasize the clinical value of mNGS as a supplement for brain abscess, enabling more comprehensive and accurate pathogen identification.IMPORTANCEThe accurate identification of pathogens causing brain abscess is crucial for effective treatment and improved patient outcomes. In this 10-year retrospective study, the detection performance of conventional culture methods and metagenomic next-generation sequencing (mNGS) was compared. The study analyzed 612 patients with brain abscess and confirmed etiology in 174 cases. The results showed that culture tests predominantly identified gram-positive bacteria, while mNGS unveiled a broader diverse pathogen spectrum, particularly anaerobes. The mNGS method exhibited significantly higher overall rates of pathogen positivity both in pus and cerebrospinal fluid (CSF) samples, surpassing the culture methods. Notably, mNGS detected a significantly higher number of anaerobes in both pus and CSF samples compared to culture methods. These findings underscore the clinical value of mNGS as a supplement for brain abscess diagnosis, enabling more comprehensive and accurate pathogen identification, particularly for rare and fastidious pathogens that evade detection by conventional culture methods.

Tandem mass spectrometry-based assay for heparan-N-sulphatase in paediatric CSF: A potential pharmacodynamic biomarker for mucopolysaccharidosis type IIIA therapy

Mucopolysaccharidosis type IIIA is a lysosomal storage disorder caused by mutations in the gene coding for heparan-N-sulphatase, a crucial enzyme in the degradation of heparan sulfate. In mucopolysaccharidosis type IIIA, heparan sulfate accumulates in the lysosomes, predominantly affecting the central nervous system. It is the most common and most severe form of mucopolysaccharidosis type III, with onset typically before the age of ten years. There is an ongoing effort to develop therapies that aim at restoring enzyme function in the brain. This study introduces a novel tandem mass spectrometry method for assessing heparan-N-sulphatase activity in pediatric cerebrospinal fluid from healthy and disease individuals.Analysis of cerebrospinal fluid samples revealed marked differences in enzyme activity, with mucopolysaccharidosis type IIIA individuals exhibiting significantly reduced levels. This new method could serve as a valuable tool for evaluating the efficacy of future therapeutic interventions targeting sulphatase activity restoration in the brain.

B-045 Evaluation of the Analytical Performance of Three Sepsis Related Assays on the Atellica CI Analyzer

Abstract Background The Atellica® CI Analyzer is an automated, high-throughput integrated chemistry and immunoassay analyzer utilizing both Atellica® CH and Atellica® IM Assays. This study evaluated the analytical performance of the Atellica IM BRAHMS Procalcitonin (PCT) Assay, and the Atellica CH Revised C-Reactive Protein (RCRP) and Lactate_3 (Lac_3) Assays on the Atellica CI Analyzer. Methods Precision studies were performed according to CLSI EP05-A3 using native and contrived human serum (PCT, RCRP) or plasma and cerebrospinal fluid (Lac_3) samples. One aliquot of each sample pool was tested in duplicate in two runs per day ≥2 hours apart on each analyzer for ≥20 days. Precision studies were evaluated with one to three lots of reagent on one to two systems. Method comparison (MC) studies were performed using three reagent lots according to CLSI EP09c. Individual native and contrived human serum (PCT, RCRP) or plasma and cerebrospinal fluid (Lac_3) samples were analyzed using the Atellica IM PCT Assay on the Atellica IM Analyzer, the Atellica CH RCRP, Lac_3 Assays on the Atellica CH Analyzer and all three Atellica IM and CH Assays listed above on the Atellica CI Analyzer. Results Representative precision and MC results for each assay are listed in the table. For the three assays tested, repeatability and within-lab %CVs were &amp;lt;1.4% and &amp;lt;8.9%, respectively. Slopes determined by the Deming linear regression model were approximately equal to 1. Conclusions Evaluation of the Atellica IM PCT and Atellica CH RCRP, Lac_3 Assays using the Atellica CI Analyzer demonstrated good precision and equivalent performance compared to the same assays on their respective Atellica CH and Atellica IM Analyzers.

Downregulation of glucose-energy metabolism via AMPK signaling pathway in granulosa cells of diminished ovarian reserve patients

Glucose metabolism plays a crucial role in the function of granulosa cells (GCs) and the development of follicles. In cases of diminished ovarian reserve (DOR), alterations in these processes can impact female fertility. This study aims to investigate changes in glucose-energy metabolism in GCs of young DOR patients aged 20 to 35 years and their correlation with the onset and progression of DOR. 72 DOR cases and 75 women with normal ovarian reserve (NOR) as controls were included based on the POSEIDON and Bologna criteria. Samples of GCs and follicular fluid (FF) were collected for a comprehensive analysis involving transcriptomics, metabolomics, RT-qPCR, JC-1 staining, and flow cytometry. The study identified differentially expressed genes and metabolites in GCs of DOR and NOR groups, revealing 7 common pathways related to glucose-energy metabolism, along with 11 downregulated genes and 14 metabolites. Key substances in the glucose-energy metabolism pathway, such as succinate, lactate, NADP, ATP, and ADP, showed decreased levels, with the DOR group exhibiting a reduced ADP/ATP ratio. Downregulation of genes involved in glycolysis (HK, PGK, LDH1), the TCA cycle (CS), and gluconeogenesis (PCK) was observed, along with reduced glucose content and expression of glucose transporter genes (GLUT1 and GLUT3) in DOR GCs. Additionally, decreased AMPK pathway activity and impaired mitochondrial function in DOR suggest a connection between mitochondrial dysfunction and disrupted energy metabolism. Above all, the decline in glucose-energy metabolism in DOR is closely associated with its onset and progression. Reduced glucose uptake and impaired mitochondrial function in DOR GCs lead to internal energy imbalances, hindering the AMPK signaling pathway, limiting energy production and supply, and ultimately impacting follicle development and maturation.

B-079 Characterization of an isoelectric focusing method for detection of IgG oligoclonal banding in paired cerebrospinal fluid and serum specimens

Abstract Background Qualitative assessment of IgG-specific oligoclonal banding (OCB) in paired serum and cerebrospinal fluid (CSF) samples is useful for evaluating inflammatory conditions of the central nervous system such as multiple sclerosis. Here we characterized the performance of an IgG-specific method using isoelectric focusing on agarose gel matrix for the detection of OCB in paired serum and CSF samples. Methods Specimens of 20 paired, simultaneously collected CSF and serum patient samples were tested on Sebia (Hydrasys 2 System) provided by the National Institutes of Health (NIH) Diagnostic Laboratories. We tested these paired samples using the Helena IgG IFE-20 kit (SPIFE® Touch) method and the results were read by two independent individuals blinded to each other’s assessments. The results were compared to the NIH results for agreement. Positive Percent Agreement (PPA) and Negative Percent Agreement (NPA) were calculated. Analytical sensitivity of the Helena assay was assessed via serial dilutions of a CSF OCB positive sample (total IgG 4.5 mg/dL). Results Figure 1 shows serum and CSF pairs for the control (A), two example positive OCB patients (B &amp; D) and a negative OCB patient (C). The Helena assay was concordant with Sebia for 11/12 positive OCB patients (PPA 91.6%) and 8/8 negative subjects (NPA 100%). 20-fold was the highest dilution that remained positive. Conclusions OCB detection using paired serum and CSF samples and isoelectric focusing electrophoresis is considered the preferred method. The Helena IgG IFE-20 assay is such a method and showed high PPA and NPA concordance with the Sebia System. Thus, the Helena OCB assay can be considered an accurate and analytically sensitive method for assessing OCB in serum and CSF.

Tetraspanins, GLAST and L1CAM Quantification in Single Extracellular Vesicles from Cerebrospinal Fluid and Serum of People with Multiple Sclerosis.

Objective: This study aimed to unravel the single tetraspanin pattern of extracellular vesicles (EVs), L1CAM+ and GLAST+ EV levels as diagnostic biomarkers to stratify people with multiple sclerosis (pwMS), specifically relapsing-remitting (RRMS) and primary progressive (PPMS). Methods: The ExoView platform was used to directly track single EVs using a clinically feasible volume of cerebrospinal fluid (CSF) and serum samples. This technology allowed us to examine the patterns of classical tetraspanin and quantify the levels of L1CAM and GLAST proteins, commonly used to immunoisolate putative neuron- and astrocyte-derived EVs. Results: The tetraspanin EV pattern does not allow us to differentiate RRMS, PPMS and non-MS donors neither in CSF nor serum, but this was associated with the type of biofluid. L1CAM+ and GLAST+ EVs showed a very low presence of tetraspanin proteins. Additionally, a significant decrease in the particle count of L1CAM+ EVs was detected in L1CAM-captured spots, and L1CAM+ and GLAST+ EVs decreased in GLAST-captured spots in the CSF from PPMS subjects compared to RRMS. Interestingly, only GLAST+ EVs exhibited a lower quantity in the CSF from PPMS compared to both MS and non-MS samples. Finally, GLAST+ EVs demonstrated a medium negative and significative correlation with GFAP levels-a biomarker of MS progression, astrocyte damage and neurodegenerative processes. Conclusions: ExoView technology could track neural EV biomarkers and be potentially useful in the diagnostic evaluation and follow-up of pwMS. GLAST+ EVs might provide insights into the etiology of PPMS and could offer small windows to elucidate the molecular mechanisms behind its clinical presentation.

The utility of disc space and vertebral body specimens cell count differential for the diagnosis of native vertebral osteomyelitis: a prospective cohort study.

Diagnostic methods for native vertebral osteomyelitis (NVO) often yield inconclusive results. Image-guided spine biopsies for culture are specific but diagnose NVO in only 50% of cases. Pre-exposure to antimicrobials further reduces diagnostic yield. Our study assesses the value of neutrophil percentage in disc space fluid and vertebral body (DS/VB) samples for diagnosing NVO. Adults referred for spine biopsy at Mayo Clinic from August 2022 to September 2023 were consented and enrolled at the time of biopsy. Following routine specimen collection, the biopsy needle was rinsed in saline into an EDTA tube for cell analysis. NVO diagnosis required organism identification in spine tissue or blood and/or positive histopathology, and consistent symptoms and imaging. Sixty-eight patients were prospectively enrolled, comprising 14 with NVO and 54 with alternative diagnoses. The median biopsy sample polymorphonuclear (PMN) percentage for NVO patients was 80.5% (IQR 72.5-85.2), compared to 64.5% (IQR 54.0-69.0) for those without NVO (p < 0.001). Nine (64.3%) NVO patients received antibiotics within 10 days prior to spine biopsy. As a continuous measure, PMN differential showed a moderately strong ability in classifying NVO status with an area under ROC curve of 0.795; an optimal point on the curve of 71.5% corresponded to a sensitivity of 78.6%, specificity of 79.6%, negative predictive value of 93.5% and positive predictive value of 50.0%. PMN differential in DS/VB biopsies may serve as an effective diagnostic tool in the evaluation of patients with NVO particularly in ambiguous cases with an initially negative spine biopsy. Future efforts will aim to implement these findings within routine clinical practice.

B-204 A Real-time Probe-based PCR Assay for Detection of Pneumocystis Pneumonia

Abstract Background Pneumocystis jirovecii is an atypical opportunistic fungus with lung tropism and worldwide distribution that continues to be one of the major opportunistic pathogens causing severe Pneumocystis pneumonia (PcP) in individuals with acquired immune deficiency syndrome (AIDS) and patients with immunosuppression due to other causes. Widespread prophylaxis and treatment for P. jirovecii with sulfa drugs have decreased the incidence of PcP, but concerns have been raised about the possible emergence of P. jirovecii isolates resistant to these drugs. Point mutations in this gene have been associated with prior exposure to sulfa drugs in other microorganisms, the polymorphic positions 165 and 171, which lead to an amino acid change at positions 55 (Thr55Ala) and 57 (Pro57Ser) of the polypeptide chain, respectively. Fluorescent PCR technology can detect the nucleic acid fragments of P. jirovecii in patients' serum, alveolar perfusion fluid (BALF) and tissue samples, and further identify drug-resistant mutations, which is of great significance for early clinical and accurate initiation of antifungal therapy. MycoMDx Pneumocystis Jirovecii PCR Assay is the use of PCR-fluorescence probe method, combined with other methods to achieve the auxiliary diagnosis of P. jirovecii infection and resistance to sulfonamides (Fig. 1). Methods In this study, we did a performance evaluation of the MycoMDx Pneumocystis Jirovecii PCR Assay (CE-approved), including diagnostic sensitivity and specificity, the limit of detection (LoD), precision and stability. The above PCR assay reagents were developed and produced by Dynamiker Biotechnology (Tianjin) Co., Ltd. Results The sensitivity and specificity of the PCR assay reagents were above 90%, the LoD was 1000 copies/mL, and the precision and stability were in accordance with the requirements (CV≤5%). Conclusions The MycoMDx Pneumocystis Jirovecii PCR Assay has great clinical value by providing a rapid and reliable test to aid in early diagnosis.

A-097 Body Fluid Validation for 11 Analytes on a Chemistry Analyzer

Abstract Background Most clinical chemistry assays are intended for use in serum, plasma, urine, and/or CSF with no performance characteristics on body fluids. However, testing of body fluids can be medically necessary for certain diagnoses. Such as determining the cause of fluid buildup in peritoneal spaces, pleural effusions, whether effusions are transudates or exudates, and even diagnosing some joint disorders. To provide these medically necessary tests, the burden is on the laboratory to perform regulatory-required validations. Herein is the validation of 11 clinical chemistry analytes in several common body fluids. Methods Mixing and stability studies were performed for the following analytes on the Roche cobas c702 (Roche Diagnostics), using the methods intended for use with serum/plasma samples: Albumin, Amylase, Bilirubin, Urea Nitrogen, Cholesterol, Creatinine, Glucose, Lactate Dehydrogenase, Lipase, Protein, and Triglycerides. Six individual fluid samples were mixed with the intended use matrix in 3 ratios: 1:3, 1:1, and 3:1. The following matrices were evaluated: pleural, peritoneal, JP drainage, synovial, and pancreatic cyst fluids. Stabilities were tested for 7 days at room temperature (RT) and 2-8°C. A maximum dilution study was also performed for Amylase. Results Mixing Study: The results are shown in the table below: Stability: For most body fluids, analytes were stable for 7 days at RT and 2-8 °C. The following analytes displayed shorter stability at RT: Bilirubin (peritoneal: 2 days, pleural: 1 day, JP drainage: 1 day), Urea Nitrogen (JP drain fluid: 2 days), Lipase (peritoneal fluid 5 days), and Triglycerides (peritoneal fluid: 5 days). Lactate dehydrogenase in pleural fluid showed a stability of 5 days at RT and 2 days at 2-8 °C. Maximum Dilution A 1:505 compounded manual and automated dilution displayed a &amp;lt;7% bias. Conclusions No sample matrix interferences were observed for any of the body fluids-analyte pairs tested allowing for this medically necessary testing.

A-116 Pleural Fluid pH - Are We Collecting and Measuring It Correctly?

Abstract Background Pleural fluid pH has diagnostic, therapeutic, and prognostic implications in the management of pleural space infections and malignancy. Pleural fluid sample collection, transportation and processing are known for source of pre-analytical error which contribute to inaccuracy of measured pH with clinical implication to patient care. Some clinical practices observed locally include pleural fluid collection in sterile container for ease of other biochemistry analytes measurement in addition to pH. This study aimed to compare the differences in pleural fluid pH and other commonly ordered biochemistry analytes collected using sterile container against heparinized blood gas syringe. Methods Total 20 Pleural fluid samples were collected from patient using a procedure called thoracentesis from, with approximately 2 to 3ml of fluid directly transferred into heparinized blood gas syringe as well as in a sterile container. Both samples were then transported to Clinical Biochemistry Laboratory within an hour. Both samples were analyzed on 3 analytical systems; 1) pH on Roche COBAS b221 blood gas analyzer, 2) Glucose, Total Protein, LDH on Beckman AU System and 3) Specific Gravity on Reflectometer. Statistical analysis was performed using Analyze-It software. Results pH fluid collected in sterile container demonstrates significant high bias (absolute of 0.4 units or 5%) as compared to sample collected in heparinizes blood gas syringe. Other commonly ordered tests in fluid profile (Glucose, Total Protein, LDH and SG) demonstrate good correlation (Altman-Bland Difference Plot &amp;lt;5%) for pleural fluid collected in sterile container as compared to heparinized syringe. Conclusions This positive bias observed is likely attributed to air exposure in pleural fluid collection in sterile container as compared to heparinized syringe. The accuracy of measured pleural pH is critically dependent on the method of sample collection. This study highlights the need for a standardized protocol for pleural pH collection for accurate measurement.

A-158 Multi-site verification and performance of body fluids in-between Horiba Medical Yumizen h2500 with Sysmex XN-10 automated hematology analyzers and manual microscopy for morphology assessment

Abstract Background The analysis of body fluids plays an important role in the diagnosis and management of a wide variety of conditions. As standards for method validation and laboratory developed tests evolve, the body fluids analysis is receiving increased attention from both laboratories and regulatory bodies. We present the evaluated result of the validation and clinical performance accuracy of HORIBA Medical Yumizen H2500 analyzers as compared to SYSMEX XN-10 analyzers and to manual slide microscopy examination as a reference method for differential analysis for body fluids samples. Methods Depending on each laboratory collected procedures, synovial samples were collected with Sodium Heparin or K2EDTA or without anticoagulant and the other samples were collected with K2EDTA or without anticoagulant. As recommended, most (94%) of the synovial fluids were treated with the hyaluronidase. For the comparison of Yumizen H2500 versus XN-10 analyzers, the study was performed in 2 US sites and 1 French site collecting 390 samples totally: 138 serous fluids (Ascites, Pleural, Peritoneal, Pericardial), 169 synovial fluids and 83 CSF fluids. Samples were analyzed as soon as possible after collection (with a limit of 24 hours for serous and synovial fluids and 4 hours for CSF) and within 2 hours between both instruments. For the comparison of Yumizen H2500 versus reference method (manual microscopy), the study was performed in 2 US sites collecting 58 samples: 34 serous fluids (Ascites, Pleural, Peritoneal, Pericardial), 12 synovial fluids and 12 CSF fluids. Two qualified morphology examiners made the manual microscopy analysis and the average of the results of the 2 slide readings was used for the analysis. Results Considering all types of body fluids, the H2500/XN results are very well correlated for BFWBC (0.969), BFRBC (0.999), BFMN% (0.912), BFMN# (0.89), BFPN% (0.912), BFPN# (0.97). Similarly, the statistical analysis per body fluids type shows very good correlation. For serous type, all results are higher than 0.876, for synovial type all results are higher than 0.879 and for cerebrospinal fluids type, all results are higher than 0.788. Among all samples, less than 8 samples (&amp;lt;2%) were out of the 95% interval. The comparison Yumizen H2500 versus manual microscopy, all the correlation results (r²) are very good: 0,9705 for Polynuclear and 0,9742 for Mononuclear. The study demonstrated better results of Yumizen H2500 versus manual microscopy than versus XN-10 for BFPN% (0.9705 versus 0.912) and BFMN% (0.9742 versus 0.912). Among 58 samples, only 4 (6.9%) were out of the 95% interval for BFPN% and BFMN%. Conclusions The Yumizen H2500 results for body fluid WBC differentiation (Polynuclear and Mononuclear) are very well correlated with the results from manual slide microscopy evaluation. The comparison of the results analyzed on Yumizen H2500 and on XN does not show any significant difference.

A-281 Comparison of C-reactive Protein, Procalcitonin, and Presepsin in Blood and Synovial Fluid as Diagnostic Biomarkers for Septic Arthritis

Abstract Background Early detection and prompt treatment are crucial for managing septic arthritis (SA). However, diagnosing SA at an early stage can be challenging since the symptoms are non-specific, and there is no definitive diagnostic test available. This study aimed to investigate the potential of biomarkers as a reliable diagnostic tool for SA. Methods A prospective study was conducted to collect residual blood and synovial fluid samples from patients who underwent blood sampling and arthrocentesis for differential diagnosis due to suspected SA. The collected samples were analyzed for C-reactive protein (CRP), procalcitonin, and presepsin. The data obtained from the analysis were subjected to statistical analysis. Results From January to November 2023, 58 patients were enrolled. They comprised 24 men and 34 women with a median age of 71.1 (interquartile range, IQR 64.0-79.3) years. Twenty-one patients were diagnosed with SA and 37 with non-SA, including osteoarthritis (n=2), osteonecrosis (n=1), meniscus tear (n=1), abscess (n=1), contusion (n=2), and mechanical complications of implants (n=30). There were no significant differences in age or sex distribution between the two groups. The median blood and synovial fluid CRP levels were 4.24 and 1.85 mg/dL, respectively, in the SA group and 0.24 and 0.11 in the non-SA group. The median blood and synovial fluid procalcitonin levels were 0.0383 and 0.0403 ng/mL in the SA group and 0.0358 and 0.0488 in the non-SA group. The median blood and synovial fluid presepsin levels were 307 and 1389 pg/mL in the SA group and 192 and 959 in the non-SA group. Except for blood procalcitonin, all showed statistically significant differences between the SA and non-SA groups. In the ROC curve analysis, blood CRP displayed the highest area under the curve (AUC) of 0.795 with a 0.47 mg/dL cut-off value, 90.5% sensitivity, and 64.9% specificity. Synovial fluid CRP displayed similar results of 0.786 AUC, 0.30 mg/dL cut-off value, 85.0% sensitivity, and 71.4% specificity. Blood and synovial fluid procalcitonin had low AUCs of 0.532 and 0.659, respectively, and were insufficient to differentiate between SA and NS groups. The AUC (0.696 vs. 0.704), sensitivity (76.2% vs. 76.2%), and specificity (67.6% vs. 62.2%) of blood and synovial fluid presepsin were similar; however, synovial fluid showed a significantly higher cut-off value, 4.5 times that of blood (237 pg/mL vs. 1087 pg/mL). Conclusions Procalcitonin was not a reliable marker for distinguishing between SA and non-SA, regardless of whether it was measured in blood or synovial fluids. However, CRP and presepsin proved useful markers when measured in either blood or synovial fluid. In particular, when measuring presepsin in synovial fluid, an optimal cut-off value is needed.

Synovial vancomycin and meropenem concentrations in periprosthetic joint infection treated by single-stage revision combined with intra-articular infusion.

We aimed to determine the concentrations of synovial vancomycin and meropenem in patients treated by single-stage revision combined with intra-articular infusion following periprosthetic joint infection (PJI), thereby validating this drug delivery approach. We included 14 patients with PJI as noted in their medical records between November 2021 and August 2022, comprising eight hip and seven knee joint infections, with one patient experiencing bilateral knee infections. The patients underwent single-stage revision surgery, followed by intra-articular infusion of vancomycin and meropenem (50,000 µg/ml). Synovial fluid samples were collected to assess antibiotic concentrations using high-performance liquid chromatography. The peak concentrations of vancomycin and meropenem in the joint cavity were observed at one hour post-injection, with mean values of 14,933.9 µg/ml (SD 10,176.3) and 5,819.1 µg/ml (SD 6,029.8), respectively. The trough concentrations at 24 hours were 5,495.0 µg/ml (SD 2,360.5) for vancomycin and 186.4 µg/ml (SD 254.3) for meropenem. The half-life of vancomycin was 6 hours, while that of meropenem ranged between 2 and 3.5 hours. No significant adverse events related to the antibiotic administration were observed. This method can achieve sustained high antibiotic concentrations within the joint space, exceeding the reported minimum biofilm eradication concentration. Our study highlights the remarkable effectiveness of intra-articular antibiotic infusion in delivering high intra-articular concentrations of antibiotics. The method provided sustained high antibiotic concentrations within the joint cavity, and no severe side-effects were observed. These findings offer evidence to improve clinical treatment strategies. However, further validation is required through studies with larger sample sizes and higher levels of evidence.

Unveiling exosomes: Cutting-edge isolation techniques and their therapeutic potential.

Exosomes are one type of nanosized membrane vesicles with an endocytic origin. They are secreted by almost all cell types and play diverse functional roles. It is essential for research purposes to differentiate exosomes from microvesicles and isolate them from other components in a fluid sample or cell culture medium. Exosomes are important mediators in cell-cell communication. They deliver their cargos, such as mRNA transcripts, microRNA, lipids, cytosolic and membrane proteins and enzymes, to target cells with or without physical connections between cells. They are highly heterogeneous in size, and their biological functions can vary depending on the cell type, their ability to interact with recipient cells and transport their contents, and the environment in which they are produced. This review summarized the recent progress in exosome isolation and characterization techniques. Moreover, we review the therapeutic approaches, biological functions of exosomes in disease progression, tumour metastasis regulation, immune regulation and some ongoing clinical trials.

Enhancing Oil Recovery in Eagle Ford Shale: A Multiscale Simulation Study of Surfactant Huff ’n’ Puff Methodology

In this paper, we present a simulation case study of a surfactant huff ’n’ puff pilot in the black oil window of the Eagle Ford (EF) Shale. The target horizontal well, which had been depleted for nearly 8 years, underwent stimulation via a surfactant huff ’n’ puff treatment. The surfactant was selected through laboratory screening using reservoir rock and fluid samples. After a 17-hour injection and a 1-month shut-in period, the well’s production increased fivefold from the baseline oil rate, sustaining incremental oil production for at least 2 years. The surfactant enhances oil recovery by altering rock wettability toward a more water-wet state and moderating oil/water interfacial tension (IFT). This process is modeled by surfactant adsorption in the simulator, indicating the degree of dynamic changes in relative permeability (krl) and capillary pressure (Pc) curves. We propose a comprehensive workflow comprising three stages: development of core-scale and field-scale models, sequential model calibrations, and multiobjective optimization to integrate laboratory measurements and field data from this pilot into multiscale numerical simulations. By matching oil recoveries from imbibition experiments on the core model and field production histories on the field model, krl and Pc profiles of two extreme states, basic reservoir properties, and additional reservoir properties altered during huff ’n’ puff operations are characterized. The matched core model reproduces a 15.1% incremental oil recovery for surfactant-assisted spontaneous imbibition (SASI) process relative to pure brine imbibition process. The matched reservoir model predicts the surfactant huff ’n’ puff treatment increases the oil production by 21.9% relative to water huff ’n’ puff treatment and by 52.9% relative to primary depletion for a 4-year period. The calibrated reservoir model also serves as a base case for optimizing well operation schedules through the implementation of a multiobjective genetic algorithm. The surfactant injection rate, injection time, and well shut-in time of the base case are varied to achieve higher oil production and reduced surfactant usage. Statistical analysis of eight trade-off cases indicates that optimal well operations, compared with existing practices, frequently involve increased injection rates [16.6–18.9 barrels per minute (bpm)], shorter injection periods (10–11.3 hours), and prolonged shut-indurations (49–65 days). This workflow offers valuable insights into surfactant huff ’n’ puff treatments for unconventional reservoirs, thereby facilitating the optimization of well operations and maximizing tertiary oil recovery.

Screening For Bioactive Compounds From Microorganisms Isolated From Trus Madi Mountain, Sabah

Sixty-four soil samples and fluid samples from seven Nephenthes pitchers were obtained from the Trus Madi Mountain, Sabah

Chromosomal Abnormalities Detected by Chromosomal Microarray Analysis and Karyotype in Fetuses with Ultrasound Abnormalities.

Chromosomal microarray analysis (CMA) is a first-line test to assess the genetic etiology of fetal ultrasound abnormalities. The aim of this study was to evaluate the effectiveness of CMA in detecting chromosomal abnormalities in fetuses with ultrasound abnormalities, including structural abnormalities and non-structural abnormalities. A retrospective study was conducted on 368 fetuses with abnormal ultrasound who received interventional prenatal diagnosis at Meizhou People's Hospital from October 2022 to December 2023. Samples of villi, amniotic fluid, and umbilical cord blood were collected according to different gestational weeks, and karyotype and CMA analyses were performed. The detection rate of chromosomal abnormalities in different ultrasonic abnormalities was analyzed. There were 368 fetuses with abnormal ultrasound, including 114 (31.0%) with structural abnormalities, 225 (61.1%) with non-structural abnormalities, and 29 (7.9%) with structural combined with non-structural abnormalities. The detection rate of aneuploidy and pathogenic (P)/likely pathogenic (LP) copy number variations (CNVs) of CMA in fetuses with structural abnormalities was 5.26% (6/114), the detection rate of karyotype was 2.63% (3/114), and the additional diagnosis rate of CMA was 2.63%. In the fetuses with ultrasonic non-structural abnormalities, the detection rate of karyotype was 6.22% (14/225), the detection rate of aneuploidy and P/LP CNVs in fetuses with ultrasonic structural abnormalities was 9.33% (21/225), and the additional diagnosis rate of CMA was 3.11%. There was no significant difference in chromosome abnormality detection rate of CMA among structural abnormality, non-structural abnormality, and structural abnormality combined with non-structural abnormality groups (5.3%, 9.3%, and 13.8%, p = 0.241), also among multiple ultrasonic abnormality and single ultrasonic abnormality groups (14.8%, and 7.3%, p = 0.105). CMA can significantly improve the detection rate of genetic abnormalities in prenatal diagnosis of ultrasonic abnormal fetuses compared with karyotype analysis. CMA is a more effective tool than karyotyping alone in detecting chromosomal abnormalities in fetuses with ultrasound abnormalities.

Neuro-Behcet's Masquerading as Status Epilepticus and Meningoencephalitis in the Emergency Department

BackgroundBehcet disease (BD) is a rare small vessel vasculitis that commonly manifests as recurrent painful oral or genital ulcerations, uveitis, and skin lesions. Some patients with BD develop neurological symptoms termed neuro-Behcet's disease. In the emergency department (ED) setting, these symptoms can be mistaken for other common acute issues including stroke, infection, epilepsy, multiple sclerosis, toxin ingestion, or psychiatric conditions. Case ReportWe present a case of a 28-year-old male with neuro-Behcet disease mimicking status epilepticus and meningoencephalitis. He was actively seizing on arrival and febrile at 103.8 F. The patient also had a history of vasculitis, uveitis and genital lesions which raised suspicion for a rheumatological process. Cerebrospinal fluid (CSF) samples were obtained and computed tomography (CT) of the head revealed no acute findings. The patient was started on treatment for seizure and meningoencephalitis and admitted to neurology for continuous electroencephalogram (EEG) monitoring with additional consults placed for rheumatology, ophthalmology, and infectious disease. The patient was ultimately diagnosed with neuro-Behcet's disease and treated appropriately.Why should an emergency physician be aware of this?: In a patient presenting with neurological symptoms and signs of rheumatological disease providers should consider neuro-Behcet's disease as a differential diagnosis. This case exemplifies the vital role of reviewing past medical history to expand differential diagnoses and early coordination with specialists so treatment can be initiated early to prevent morbidity and mortality. The case will also review different presentations of this unfamiliar diagnosis for a more comprehensive understanding.