Fluid Analysis Research Articles

Biochemical analysis of soft tissue infectious fluids and its diagnostic value in necrotizing soft tissue infections: a 5-year cohort study

BackgroundNecrotizing soft tissue infections (NSTI) are rapidly progressing and life-threatening conditions that require prompt diagnosis. However, differentiating NSTI from other non-necrotizing skin and soft tissue infections (SSTIs) remains challenging. We aimed to evaluate the diagnostic value of the biochemical analysis of soft tissue infectious fluid in distinguishing NSTIs from non-necrotizing SSTIs.MethodsThis cohort study prospectively enrolled adult patients between May 2023 and April 2024, and retrospectively included patients from April 2019 to April 2023. Patients with a clinical suspicion of NSTI in the limbs who underwent successful ultrasound-guided aspiration to obtain soft tissue infectious fluid for biochemical analysis were evaluated and classified into the NSTI and non-necrotizing SSTI groups based on their final discharge diagnosis. Common extravascular body fluid (EBF) criteria were applied.ResultsOf the 72 patients who met the inclusion criteria, 10 patients with abscesses identified via ultrasound-guided aspiration were excluded. Based on discharge diagnoses, 39 and 23 patients were classified into the NSTI and non-necrotizing SSTI groups, respectively. Biochemical analysis revealed significantly higher albumin, lactate, lactate dehydrogenase (LDH), and total protein levels in the NSTI group than in the non-necrotizing SSTI group, and the NSTI group had significantly lower glucose levels and pH in soft tissue fluids.In the biochemical analysis, LDH demonstrated outstanding discrimination (area under the curve (AUC) = 0.955; p < 0.001) among the biochemical markers. Albumin (AUC = 0.884; p < 0.001), lactate (AUC = 0.891; p < 0.001), and total protein (AUC = 0.883; p < 0.001) levels also showed excellent discrimination. Glucose level (AUC = 0.774; p < 0.001) and pH (AUC = 0.780; p < 0.001) showed acceptable discrimination. When the EBF criteria were evaluated, the total scores of Light’s criteria (AUC = 0.925; p < 0.001), fluid-to-serum LDH ratio (AUC = 0.929; p < 0.001), and fluid-to-serum total protein ratio (AUC = 0.927; p < 0.001) demonstrated outstanding discrimination.ConclusionBiochemical analysis and EBF criteria demonstrated diagnostic performances ranging from acceptable to outstanding for NSTI when analyzing soft tissue infectious fluid. These findings provide valuable diagnostic insights into the recognition of NSTI. Further research is required to validate these findings.

Comparative data analysis of radiative MHD Bingham and Newtonian fluid past a Non-Linear stretching sheet with thermoelectric and Joule effects

Comparative data analysis of radiative MHD Bingham and Newtonian fluid past a Non-Linear stretching sheet with thermoelectric and Joule effects

P39 Primary antiphospholipid syndrome before development of severe systemic lupus erythematosus – a case report

Abstract Introduction Antiphospholipid syndrome (APS) has long been associated with systemic lupus erythematosus (SLE). Graham Hughes first reported APS in 1983 in cohorts of SLE patients with recurrent thrombosis. We present a case of a young man who developed severe manifestations of SLE 7 years after the diagnosis of primary APS and we look at the literature for predictors in primary APS to develop into SLE. Case description A 28 year old Caucasian male with Turkish ancestry was diagnosed with triple positive primary APS at the age of 21 when he presented with bilateral pulmonary embolisms. He was stable on warfarin with target INR between 3-4. He had been having occasional mouth ulcers, lethargy, mild paraesthesia of his feet and headaches for 2 months. He then presented in February 2024 with short history of pleuritic chest pain and marked breathlessness. Initial cross sectioning imaging did not reveal pulmonary embolism. A week later, echocardiogram demonstrated large global pericardial effusion and small pleural effusions. Ejection fraction was preserved. Interval MRI scan showed pericarditis without evidence of myocarditis. Almost 600 mls of heavily blood stained pericardial fluid was drained. Analysis of the pericardial fluid showed inflammatory processes with macrophages, neutrophil and lymphocytes. Culture was negative including for tuberculosis and cytology negative for malignancy. Immunology revealed ANA 1:160, ENA normal, dsDNA was initially negative but upon interval testing was high at 27.5 IU/mL, IgG 13.1 g/L, C3 1.2 g/L, C4 &amp;lt;0.1 g/L, CRP 143, ESR 30. He had anaemia, lymphopenia and positive direct antiglobulin test. ANCA and rheumatoid factor were unremarkable. Renal and joints were unaffected. Due to his Turkish ancestry, he was extensively investigated for the possibility of Behcet’s with normal femoral vein ultrasound and negative HLA-B51 genotype. His manifestations fulfilled the SLICC criteria to be diagnosed with SLE. His response to intravenous methylprednisolone infusions and subsequent oral steroid was marked. He was initiated on dual treatment of hydroxychloroquine and mycophenolate mofetil and is currently recovering. Discussion We relooked at this immunology when he was first diagnosed with primary APS in 2017. He was strongly positive for ANA at titre of 1:640 but did not have manifestations of SLE then. We researched literature on the predictors for primary APS to develop into SLE. Both disease spectrums have several overlapping manifestations including haemolytic anaemia, thrombocytopenia, lymphopenia, neurological symptoms, renal manifestations and livedo reticularis. Freire et al (1) retrospectively analysed 80 patients with primary APS and found 14 (17.5%) progressed to develop APS-SLE in 5.2 +/- 4 years. ANA positivity was found in 100% those who eventually developed APS-SLE. Other specific autoantibodies including anti-dsDNA, anti-ribosomal P, anti-Ro, anti-La and anti-U1RNP are found more commonly in the cohort too. In their APS-SLE cohort, they also found a correlation with younger age at the time of diagnosis of APS and the duration of the disease. Andreoli L et al (2) demonstrated anti-nucleosome antibody (anti-NCS) was frequently found in their cohort of APS-SLE patients but no relationship between clinical-serological was found, except for a weak correlation between anti-dsDNA antibodies. 2 of their primary APS who developed severe SLE had displayed high titres of anti-NCS years before. Tarr T et al (3) found 4 of their primary APS who developed SLE all presented with cerebrovascular events as the thrombotic manifestation. Key learning points • Primary APS has been associated with the development of SLE • Certain antibody profile such as ANA, anti-nucleosome antibody may predict the development of SLE in primary APS patients • Younger age, cerebrovascular events as thrombotic manifestation at diagnosis of primary APS and the length of duration of primary APS may be associated with the development of SLE at a later stage

Proteomic Analysis of Follicular Fluid in Polycystic Ovary Syndrome: Insights into Protein Composition and Metabolic Pathway Alterations.

This study explores the proteomic composition of follicular fluid (FF) from women undergoing oocyte retrieval for in vitro fertilisation (IVF), with a focus on the effects of polycystic ovary syndrome (PCOS). FF samples were collected from 74 patients, including 34 with PCOS and 40 oocyte donors. Proteomic profiling using machine learning identified significant differences in protein abundance between the PCOS and control groups. Of the 484 quantified proteins, 20 showed significantly altered levels in the PCOS group. Functional annotation and pathway enrichment analysis pointed to the involvement of protease inhibitors and immune-related proteins in the pathophysiology of PCOS, suggesting that inflammation and immune dysregulation may play a key role. Additionally, HDL assembly was identified as a significant pathway, with apolipoprotein-AI (APOA1) and alpha-2-macroglobulin (A2M) as the major proteins involved. Notably, myosin light polypeptide 6 was the most downregulated protein, showing the highest absolute fold change, and may serve as a novel independent biomarker for PCOS.

Evaluation of Ascitic Fluid Cytology in a Tertiary Hospital in South-Western Nigeria: A 7-year Retrospective Study

Background: Ascites is the pathological accumulation of fluid within the peritoneal cavity of varied etiology. Various abnormalities can be detected through cytological analysis of ascitic fluid. Ascitic fluid cytology (AFC) is a rapid and cost-effective means of evaluating ascites to establish its nature and cause. Aim: To evaluate ascitic fluid cytological findings over a seven-year period and to correlate the cytological features with the clinical profile of the patients. Materials and Methods: This was an observational retrospective study conducted in a healthcare facility in south-western Nigeria. The reports of AFC tests performed from January 1, 2016 to December 31, 2022 were retrieved to obtain the following information for analysis: Age, Gender, Tribe, Year of the test, Provisional diagnosis, and Cytological diagnosis. Results: The data of a total number of 121 patients were analyzed; 61 males (50.4%) and 60 females (49.6%) with a mean (±SD) age of 51.51 ± 15.22. Decompensated cirrhosis was the commonest clinical diagnosis 65 (53.7%) followed by hepatocellular carcinoma 11 (9.1%). Ascitic fluid cytological findings showed 45 (37.2%) were positive for malignant cells; 46 (38.0%) were negative for malignant cells; 13 (10.7%) were suspicious for malignancy; 9 (7.4%) were acellular smears; and 8 (6.6%) were inflammatory smears. The ascitic fluid of only 4 out of 11 patients with hepatocellular carcinoma tested positive for malignant cells, giving a sensitivity of 36.4%. Conclusion: AFC remains very useful in the diagnostic work-up of patients with ascites but should not be completely relied upon in confirming a malignancy, other additional diagnostic investigations should be deployed.

Evaluation of Brain and Spinal Cord Lesions and Cerebrospinal Fluid Analysis in Detecting Demyelinating Diseases in Patients with Optic Neuritis

ABSTRACT Optic neuritis can be an early sign of demyelinating diseases like multiple sclerosis (MS), neuromyelitis optica spectrum disorders (NMOSD), and myelin oligodendrocyte glycoprotein (MOG) antibody-associated diseases (MOGAD). We investigated the presence or absence of head and spinal cord lesions on magnetic resonance imaging (MRI) and assessed whether cerebrospinal fluid (CSF) tests are useful in detecting demyelinating disease in patients with first diagnosed optic neuritis. We conducted a retrospective study of 111 patients (47 idiopathic, 19 NMOSD, 16 MOGAD, 16 MS, 6 optic neuritis with cerebral lesions but that does not meet the McDonald’s criteria for MS (ON+)), and 7 chronic relapsing inflammatory optic neuropathy) diagnosed with optic neuritis without cerebral or spinal symptoms. Patients underwent evaluations including orbital, head, and spine MRI, along with CSF analysis. Among the 111 patients, 20 (35.1%: 4 NMOSD, 4 MOGAD, 7 MS, and 6 ON+) exhibited intracerebral or spinal cord lesions. Twelve patients showed findings on both orbital and head MRI, while six had no orbital MRI findings except for optic neuritis but exhibited lesions on head MRI. Five patients had spinal lesions without intracerebral lesions. CSF analysis revealed positive oligoclonal bands and elevated myelin basic protein levels indicate the high likelihood with systemic inflammatory demyelinating diseases. Even in the absence of concomitant encephalitis or myelitis symptoms or a history of these conditions, MRI images of patients with optic neuritis sometimes reveal lesions in the brain or spinal cord. CSF abnormalities were indicative of systemic demyelinating disease presence, extending beyond MS to NMOSD and MOGAD.

Musculoskeletal Ultrasound Characteristics of Checkpoint Inhibitor-Associated Inflammatory Arthritis

BackgroundCancer immunotherapy with checkpoint inhibition (ICI) has revolutionized the treatment of solid cancers; however, it is associated with a spectrum of immune-related adverse events (irAEs), including inflammatory arthritis. Here we report our experience with the use of point-of-care musculoskeletal ultrasound (MSKUS) and provide a description of MSKUS findings in patients with definite musculoskeletal irAEs. MethodsPatients ≥18 years who received ICI at the Mass General Cancer Center from 2010-2019 were referred to rheumatology by oncology for evaluation of musculoskeletal symptoms following ICI therapy. Fifty-five patients with suspected MSK irAEs had MSKUS performed and interpreted by the same ultrasonographer. Findings were reviewed and confirmed by a blinded US reader. US findings in patients with definite de novo MSK irAEs were reviewed and correlated with the presence or absence of documented clinical synovitis and with available synovial fluid analysis. ResultsThirty-four out of fifty-five patients (62%) had definite de novo irAE. Seven patients were identified with alternative etiologies assisted by diagnostic MSKUS. Twenty patients with definite de novo irAE had clinical evidence of synovitis at the time of the initial MSKUS examination, while 14 did not. Among patients with clinically evident synovitis, MSKUS examination confirmed inflammatory pathology in all patients. The most common MSKUS features identified were grade 2 or higher synovial thickening (80%), hyperemia measured by color power Doppler (CPD) signal (70%), and tenosynovitis (60%). Among the 14 patients without clinically evident synovitis, inflammatory features were identified in 10 patients (71%); the most common features identified were > grade 1 synovial proliferation, hyperemia and tenosynovitis. Of 15 patients who underwent synovial fluid analysis, 7 patients had synovial fluid cell counts < 2000 cells/µL considered traditionally within the ‘non-inflammatory’ range, and all 7 patients were noted to have inflammatory MSKUS findings. ConclusionPoint-of-care MSKUS is a valuable tool in the evaluation of potential MSK irAEs. Our data demonstrates its ability to expediate early identification of subclinical synovitis and/or tenosynovitis even when synovial fluid analysis is within the traditional non-inflammatory range.

Aquaporin-4 antibody-positive neuromyelitis optica spectrum disorder triggered by herpes zoster: a case report and literature review.

This paper presents the case of a 45-year-old woman who developed aquaporin-4 (AQP4) antibody-positive neuromyelitis optica spectrum disorder (NMOSD) approximately 6 weeks after a herpes zoster infection. Her initial symptoms included area postrema syndrome, which was marked by persistent nausea, vomiting, and belching. This report also provides a summary of the demographic and clinical features, disease progression, magnetic resonance imaging findings, cerebrospinal fluid analysis, treatment plans, and recovery outcomes of 11 patients-including this case and 10 others reported in the literature since 2008-who developed AQP4 antibody-positive NMOSD following herpes zoster infection. This review aims to improve clinicians' understanding of the characteristics, treatment, and prognosis of this disease. It also highlights the importance for pain management specialists to consider central pain and use the MIDNIGHTS or VINDψCATE mnemonic devices to systematically consider the differential diagnoses of a patient's new pain symptoms.

Peritoneal Leak Causing Right Sided Hydrothorax in A Case of Chronic Kidney Disease Undergoing Continuous Ambulatory Peritoneal Dialysis: A Case Report From Nepal

ABSTRACTContinuous ambulatory peritoneal dialysis (CAPD) is used for the maintenance dialysis in patients with end stage renal disease. Treatment's complications in CAPD generally exist, of which transudate hydrothorax falls under the minority of the cases. Here, we aim to present a case of a 51 years old female, a newly diagnosed case of chronic kidney disease (CKD) undergoing CAPD for 3 weeks who presented at our institution with complaints of difficulty in breathing. The symptom had gradually progressed after her recent CAPD session. Several investigations were sought out, of which a chest X‐ray illustrated a right sided hydrothorax. Pleural fluid analysis showed a transudate characteristic, and other diagnostic tests suggested that a peritoneal fluid leak might have caused the hydrothorax. CAPD induced hydrothorax is one of the uncommon noninfectious complications associated with peritoneal dialysis; its diagnosis can be made by studying both biochemical and radiological parameters of the patient. The definitive diagnosis is made by establishing a pleuro‐peritoneal communication using peritoneal scintigraphy. An immediate halt in CAPD will allow the remission of hydrothorax as mentioned below in our case report. The definitive management includes video assisted thorascopic surgery with mechanical pleurodesis. An appropriate diagnostic and management approach should be sought in CAPD induced transudate hydrothorax. An immediate halt in CAPD with surgical interventions can be considered as a treatment options for such complication.

Treatment with Leflunomide in Conjunction with Glucocorticoids for Dogs with Immune-Mediated Polyarthritis Is Not Associated with Improved Outcomes: A Retrospective Cohort Study of 93 Dogs from Australia (2017–2024)

Immune-mediated polyarthritis (IMPA) has a relatively high relapse rate compared to other immune-mediated diseases. Leflunomide is frequently used to treat dogs with IMPA in conjunction with prednisolone. This retrospective cohort study aimed to evaluate the therapeutic efficacy of leflunomide as an adjunctive therapy to prednisolone in reducing relapse and mortality rates in dogs diagnosed with IMPA in Australia. The medical records of client-owned dogs diagnosed with IMPA at a specialist referral hospital in Southeast Queensland from 2017 to 2024 were reviewed. A total of 93 dogs were included in this study, divided into two groups based on the treatment received: Group PRED, consisting of 53 dogs treated with prednisolone as the sole immunosuppressive agent, and Group L+PRED, consisting of 40 dogs that received leflunomide as adjunctive therapy alongside prednisolone. Data collected included breed, age, weight, sex, serum C-reactive protein concentration, results of synovial fluid analysis and microbial culture, treatment protocol, relapse rates and time to relapse, and mortality rates. There was no difference in relapse or mortality rates, time to relapse, nor time to discontinue prednisolone between the PRED and L+PRED groups. The L+PRED group had higher body weights and lower prednisolone dose rate at discharge compared to those in the PRED group. This study demonstrated that the use of leflunomide as an adjunctive therapy to prednisolone for the treatment of dogs with IMPA had no improved outcomes, reduced relapse rates, or shortening in the duration of prednisolone therapy when compared to dogs receiving prednisolone monotherapy.

Epstein–Barr Virus encephalitis associated hemophagocytic lymphohistiocytosis in childhood-onset systemic lupus erythematosus: a case-based review

BackgroundHemophagocytic lymphohistiocytosis (HLH) is characterized by immune dysregulation that results in an uncontrolled hyperinflammatory state. HLH is classified into two main categories: primary (familial) HLH and secondary (acquired) HLH. Secondary HLH can result from various underlying, including infection-associated hemophagocytic syndrome (IAHS) and macrophage activation syndrome (MAS) associated with rheumatologic disorders, among others. Epstein–Barr virus (EBV) often causes IAHS, but central nervous system (CNS) involvement is rare among systemic lupus erythematosus (SLE) patients. We report a case of EBV encephalitis associated with HLH in a patient with childhood-onset SLE.Case presentationA 12-year-old girl had received a diagnosis of SLE 2 months before presentation. After a period of inactive disease on treatment, fever and seizures, with altered mental status and hallucinations, developed over several weeks. A complete blood cell count (CBC) revealed pancytopenia, accompanied by elevated levels of inflammatory markers: 86 mm/hr erythrocyte sedimentation rate, 8.9 mg/dl c-reactive protein, and 3,966 ng/mL of ferritin. The differential diagnosis included active neuropsychiatric SLE, CNS infection and neurological manifestations in secondary HLH, which could have represented either IAHS or MAS. Meropenem and acyclovir were initially administered for clinical acute encephalitis, followed by pulse methylprednisolone; however, the fever persisted, and another CBC revealed progressive cytopenia. A bone marrow study showed hypocellularity and active hemophagocytic activity, and intravenous immunoglobulin was additionally given due to the diagnosis of HLH. Cerebrospinal fluid (CSF) analysis showed 60/mm3 white blood cells (N 55%, L 45%), 141 mg/dL glucose (0.7 blood-CSF glucose ratio), < 4 mg/dL protein; results of Gram stain and bacterial culture were negative. The viral encephalitis panel from the CSF confirmed EBV infection. Bone marrow immunohistochemistry examination revealed increasing levels of CD8 + T-cell and equivocal positive results for EBV-encoded RNA in situ hybridization; therefore, HLH potentially associated with EBV was diagnosed. After treatment with IVIg, cyclosporin A, and prednisolone, the patient’s symptoms gradually improved and she was eventually able to return to school.ConclusionsOur case highlights the importance of a thorough differential diagnosis, including EBV encephalitis associated with HLH, in patients with childhood SLE, particularly in cases of clinical deterioration occurs after initial treatment.

Neurological dysfunction and fatal encephalitis due to Halicephalobus gingivalis in two horses in Belgium

SummaryTwo horses kept on different farms at a geographical distance of 150 km were presented with acute neurological dysfunction. Ante‐mortem diagnostic tests including bloodwork, CT scan and cerebrospinal fluid analysis did not provide a diagnosis in either case. Due to rapid deterioration, both horses were euthanised shortly after admission. Postmortem histopathological examination revealed granulomatous meningoencephalitis with intralesional nematodes that were morphologically identified as Halicephalobus gingivalis. This is the first documentation of central nervous system infestation by H. gingivalis in horses residing in Belgium, emphasising the importance of considering aberrant parasitic migration in the differential diagnosis of acute neurological dysfunction.

Label-Free Quantitative Proteomics Analysis of Nasal Lavage Fluid in Chronic Rhinosinusitis with Nasal Polyposis

(1) Background: Chronic rhinosinusitis (CRS) is a common chronic inflammation of the nasal mucosa and the paranasal sinuses. The pathogenesis of chronic rhinosinusitis (CRS) is multifactorial and, as of yet, not well understood. (2) Methods: Nasal lavage fluid samples were collected from patients diagnosed with chronic sinusitis with nasal polyposis (CRSwNP) (n = 10) and individuals without sinusitis (control group) (n = 10) who had no nasal complaints. In the present study, we used an untargeted label-free LC-MS/MS mass spectrometric approach combined with bioinformatics and network pathway analysis to compare the changes in the proteomic profiles of the CRSwNP group and the control group. Data from LC-MS/MS underwent univariate and multivariate analyses. (3) Results: The proteomic analyses revealed distinct differences in the abundances of nasal lavage fluid proteins between the CRSwNP and control groups: a total of 234 proteins, 151 up- and 83 down-regulated in CRSwNP. Functional Gene Ontology (GO) analysis showed that dysregulated proteins were involved in airway inflammatory reaction, immune response, and oxidative stress. The biomarkers were evaluated using the Receiver Operating Characteristic (ROC) curve; an Area Under the Curve (AUC) of 0.999 (95% CI) identified potential biomarkers between the CRSwNP and control group. EMILIN-3 and RAB11-binding protein RELCH were down-regulated, and Macrophage migration inhibitory factor and deoxyribonuclease-1 were up-regulated, in CRSwNP compared to the control group. (4) Conclusions: These differentially expressed proteins identified in CRSwNP are involved in airway inflammatory reaction, immune response, and oxidative stress. In particular, the identification of increased interleukin-36 gamma (IL-36γ), which contributes to inflammatory response, and a decrease in SOD, in this group are notable findings. In the future, several of these proteins may prove useful for exploring the pathogenesis of nasal polyps and chronic sinusitis or as objective biomarkers for quantitatively monitoring disease progression or response to therapy.

‘When nature breaks the rules’: Guillain-barré syndrome presenting with descending paralysis

Guillain-Barré Syndrome (GBS) typically presents with ascending paralysis, but we report an atypical case involving descending paralysis in a 36-year-old male. The patient experienced upper limb weakness and respiratory muscle involvement, progressing to respiratory distress. Initial cerebrospinal fluid (CSF) analysis and nerve conduction studies (NCS) were normal, delaying the diagnosis. Treatment with intravenous immunoglobulin (IVIG) led to partial recovery. This case highlights the diagnostic challenges of atypical GBS presentations and the importance of early intervention.

Datura stramonium seed ingestion leading to unintentional poisoning in a 3-year-old Ethiopian toddler: case report

IntroductionDatura stramonium poisoning, resulting from the ingestion of seeds, leaves, or flowers of Datura stramonium, is a severe condition with significant risks, particularly for young children. Most documented cases of Datura stramonium poisoning in the literature involve teenagers who intentionally ingest the plant to induce hallucinogenic and euphoric experiences. This report presents a rare instance of unintentional Datura stramonium poisoning in a 3year-old Ethiopian toddler.Case presentationHe presented with symptoms consistent with anticholinergic toxidrome, including altered mental status, agitation, seizures, dilated pupils, and tachycardia. Laboratory tests, including complete blood count, liver function test, renal function test, electrolyte levels and cerebrospinal fluid analysis were normal. Results of blood toxicology screen were negative. Timely recognition and supportive care lead to a positive outcome.ConclusionThrough this report, we aim to add to the limited body of literature on Datura stramonium poisoning in toddlers and offer insights into its clinical course and management in paediatric patients.

Progranulin derivative attenuates lung neutrophilic infiltration from diesel exhaust particle exposure.

Air pollutants, such as diesel exhaust particles (DEPs), induce respiratory disease exacerbation with neutrophilic infiltration. Progranulin (PGRN), an epithelial cell and macrophage-derived secretory protein, is associated with neutrophilic inflammation. PGRN is digested into various derivatives at inflammatory sites and is involved in several inflammatory processes. PGRN and its derivatives likely regulate responses to DEP exposure in allergic airway inflammation. To investigate the role of PGRN and its derivatives in the regulation of responses to DEP exposure in allergic airway inflammation. A murine model of allergic airway inflammation was generated in PGRN-deficient mice, and they were simultaneously exposed to DEP followed by intranasal administration of full-length recombinant PGRN (PGRN-FL) and a PGRN-derived fragment (FBAC). Inflammatory status was evaluated by bronchoalveolar lavage fluid and histopathologic analyses. Human bronchial epithelial cells were stimulated with DEPs and house dust mites (HDMs), and the effect of FBAC treatment was evaluated by assessing various intracellular signaling molecules, autophagy markers, inflammatory cytokines, and intracellular oxidative stress. DEP exposure exaggerated neutrophilic inflammation, enhanced IL-6 and CXCL15 secretions, and increased oxidative stress in the murine model; this effect was greater in PGRN-deficient mice than in wild-type mice. The DEP-exposed mice with PGRN-FL treatment revealed no change in neutrophil infiltration and higher oxidative stress status in the lungs. On the contrary, FBAC administration inhibited neutrophilic infiltration and reduced oxidative stress. In human bronchial epithelial cells, DEP and HDM exposure increased intracellular oxidative stress and IL-6 and IL-8 secretion. Decreased nuclear factor erythroid 2-related factor 2 (Nrf2) expression and increased phosphor-p62 and LC3B expression were also observed. FBAC treatment attenuated oxidative stress from DEP and HDM exposure. FBAC reduced neutrophilic inflammation exaggerated by DEP exposure in a mouse model of allergic airway inflammation by reducing oxidative stress. PGRN and PGRN-derived proteins may be novel therapeutic agents in attenuating asthma exacerbation induced by air pollutant exposure.

Immunological characteristics of bronchoalveolar lavage fluid and blood across connective tissue disease-associated interstitial lung diseases.

Interstitial lung disease (ILD) is a serious complication of connective tissue diseases (CTDs). The heterogeneity of ILDs reflects differences in pathogenesis among diseases. This study aimed to clarify the characteristics of CTD-ILDs via a detailed analysis of the bronchoalveolar lavage fluid (BALF) and blood immune cells. BALF and blood samples were collected from 39 Japanese patients with newly diagnosed ILD: five patients with Sjögren's syndrome (SS), eight patients with dermatomyositis (DM), six patients with rheumatoid arthritis (RA), six patients with systemic sclerosis, four patients with anti-neutrophil cytoplasmic antibody-associated vasculitis, and 10 patients with idiopathic interstitial pneumonia. We performed single-cell RNA sequencing to analyze the gene expression profiles in these patients' immune cells. In patients with SS, B cells in the BALF were increased and genes associated with the innate and acquired immunity were enriched in both the BALF and blood. In contrast, patients with DM showed an upregulation of genes associated with viral infection in both the BALF and blood. In patients with RA, neutrophils in the BALF tended to increase, and their gene expression patterns changed towards inflammation. These disease-specific characteristics may help us understand the pathogenesis for each disease and discover potential biomarkers.

A view of immunologic male infertility from the perspective of preserving seminal fluid homeostasis

The basis of the pathogenesis of immunologic male infertility is the production of antisperm antibodies against sperm antigens by plasmocytes. Extracellular nucleic acids, among other antigens, are perceived as DAMPs. The clearance system for biological fluids is complex. Endonucleases are responsible for the utilization of extracellular DNA. Among endonucleases, DNase 1 is the most studied. Its concentration in biological fluids is higher than others. For the first time, we determined the concentration of DNase 1 in the seminal fluid of men of reproductive age. DNase 1 concentration values were compared with the level of antisperm antibodies in the ejaculate, after which the relationship between the enzyme level and standard spermogram parameters was determined. The material for the study was the ejaculate of 44 relatively healthy men of reproductive age. During spermiological analysis, macroscopic and microscopic parameters of seminal fluid were determined, the concentration of deoxyribonuclease 1 (DNase 1) in the material was determined by enzyme immunoassay, and the presence of antisperm antibodies was assessed using a direct diagnostic method – the MAR test. Statistical processing of the obtained data was carried out using analytical software PAST v. 4.06. Reference intervals for DNase 1 concentrations were calculated using the MedCalc v. 17.4 program. The relationship between spermogram parameters and DNase 1 concentration was assessed by calculating Spearman’s linear rank correlation coefficients. As a result, a negative correlation was found between the DNase 1 ratio and such ejaculate parameters as liquefaction time (rS = -0.37; p = 0.013) and viscosity (rS = -0.37; p = 0.013), as well as the level of antisperm antibodies A (rS = -0.43; p = 0.003) and G (rS = -0.33; p = 0.027), the mucous component of the ejaculate (rS = -0.37; p = 0.012) and agglutination of male germ cells (rS = -0.33; p = 0.029). According to the data obtained, the optimal level of DNase 1 is directly related to the quality of the ejaculate. Determination of DNase 1 concentration can serve as an additional diagnostic criterion for male immunological infertility. These data will expand our understanding of a man’s fertile potential.

Salinity and oven-drying effects on the plasticity of a marine soft clay

Considering the extensive use of plasticity-based correlations in geotechnical practice to estimate soil parameters, this paper evaluates the influence of pore fluid salinity and soil drying on the plasticity of Ballina clay, an estuarine soft clay from northern New South Wales (Australia). A comprehensive experimental study, which includes controlled leaching/salinisation paths applied to natural (remoulded) as well as oven-dried clay, prior to the estimation of the Atterberg limits, is presented. Plasticity tests are complemented with chemical analysis of the pore fluid carried out to evaluate the processes involved in the leaching/salinisation mechanisms for remoulded and oven-dried clay. A strong dependency of liquid limit on pore fluid salinity and oven-drying is observed in Ballina clay. Leaching modifies the soil fabric from an initially saline–sodic flocculated towards a normal flocculated arrangement. The experimental results show that changes in soil plasticity upon leaching are largely reversible upon salinisation paths. Oven-drying promotes the stacking of clay minerals (aggregation), which in turn reduces the water absorption capacity of the clay. The consequences of neglecting both salinity and drying effects in practice when adopting well-established relationships between mechanical parameters and soil plasticity are also briefly discussed.

Proteomic and Targeted Lipidomic Analyses of Fluid and Rigid Rubber Particle Membrane Domains in Guayule.

Rubber (cis-1,4-polyisoprene) is produced in cytosolic unilamellar vesicles called rubber particles (RPs), and the protein complex responsible for this synthesis, the rubber transferase (RTase), is embedded in, or tethered to, the membranes of these RPs. Solubilized enzyme activity is very difficult to achieve because the polymerization of highly hydrophilic substrates into hydrophobic polymers requires a polar/non-polar interface and a hydrophobic compartment. Using guayule (Parthenium argentatum) as a model rubber-producing species, we optimized methods to isolate RP unilamellear membranes and then a subset of membrane microdomains (detergent-resistant membranes) likely to contain protein complexes such as RTase. The phospholipid and sterol composition of these membranes and microdomains were analyzed using thin-layer chromatography (TLC) and liquid chromatography tandem mass spectroscopy (LC-MS/MS). Our data indicate that RP membranes consist predominantly of phosphatidic acid-containing membrane microdomains (DRMs or "lipid rafts"). Proteomic analyses of guayule RP membranes and membrane microdomains identified 80 putative membrane proteins covering 30 functional categories. From this population, we have tentatively identified several proteins in multiple functional domains associated with membrane microdomains which may be critical to RTase function. Definition of the mechanisms underlying rubber synthesis will provide targets for both metabolic engineering and breeding strategies designed to increase natural rubber production in latex-producing species.