Genetic signatures have added a molecular dimension to prognostics and therapeutic decision-making. However, tumour heterogeneity in prostate cancer and current sampling methods could confound accurate assessment. Based on previously published spatial transcriptomic data from multifocal prostate cancer, we created virtual biopsy models that mimic conventional biopsy placement and core size. We then analysed the gene expression of different prognostic signatures  using a step-wise approach with increasing resolution from pseudo-bulk analysis of the whole biopsy, to differentiation by tissue subtype , followed by distinct tumour grade and finally clonal resolution. The gene expression profile of virtual tumour biopsies revealed clear differences between grade groups and tumour clones, compared to a benign control, which were not reflected in bulk analyses. This suggests that bulk analyses of whole biopsies or tumour-only areas, as used in clinical practice, may provide an inaccurate assessment of gene profiles. The type of tissue, the grade of the tumour and the clonal composition all influence the gene expression in a biopsy. Clinical decision making based on biopsy genomics should be made with caution while we await more precise targeting and cost-effective spatial analyses.The online version contains supplementary material available at 10.1186/s12943-023-01863-2. Prostate cancer (PCa) progression is unpredictable. Some tumours are indolent, whilst others tumours are aggressive with rapid progression and metastasis . Recent spaceranger aggr (v2.0). Fold changes with false discovery rate were analysed using EdgeR (v3.40.1).Gene expression analysis was performed on radical prostatectomy tissue from a patient with multifocal PCa. We used our recently published organ-wide ST data  to constWe compared the gene expression of virtual tumour biopsies to a benign control at increasing levels of resolution: from whole biopsy, through tissue subtype  and tumour grade, to clonal level Fig. . A conseWe observed clear evidence of variation in the expression of constituent genes from different prognostic signatures within biopsies Fig. . In tumoPreviously , we inte). Similar to Biopsy 1, this information would be missed if either the whole biopsy, or tumour areas alone, were analysed in bulk. We obtained consistent results when we extended these analyses to the other genomic scores. As with biopsy 1, we also observed differences between tumour clones  and the stromal response markers BGN, COL1A1) decreased in GG2/GG4 areas compared to the benign control . Conversely, these genes demonstrated an inverse profile if the biopsy was analysed in bulk   , we proc_1 , we pThe effectiveness of prognostic genomic tests depends on accurately deciphering the genomic landscape of a heterogeneous and unpredictable cancer for each patient , 10. InaWe are not the first to highlight this problem. Other studies have reported the confounding effects of heterogeneity in genomic tests , 11, 12.We recognise the limitations of presenting data from a single patient. We also point out that there are other examples of molecular heterogeneity (e.g. point mutations and epigenetic changes) that are not captured by the ST approach. It will be interesting to replicate these analyses as we put further prostates through our organ-scale ST pipeline. Longitudinal sampling will also be valuable to inform clonal development over time. The current cost for organ-scale analysis of a single prostate is around £100,000, but as such costs fall, we anticipate developing a more comprehensive understanding of PCa heterogeneity and lethal clonality. Furthermore, the accessibility of insights derived from these spatial molecular technologies will likely improve with the integration of machine learning into morpho-molecular studies, paving the way for a new era of precision medicine.In conclusion, in this short article we show that tissue type, tumour grade, and clonal composition of tissue all influence gene expression within a biopsy sample. Bulk analyses of prostate biopsies for genomic scores used in clinical practice effectively ignore these distinctions. To maximise the potential of biopsy-based genomics in clinical decision-making, we believe that precise targeting will need to be combined with more granular spatial analyses, in order to provide accurate scores which will better-inform clinical decisions.Below is the link to the electronic supplementary material.Supplementary Material 1Supplementary Material 2Supplementary Material 3Supplementary Material 4


S. pneumonia) antigen test drawn from the pleural fluid rather than a urine sample has been found to have equivalent diagnostic utility to the urinary antigen test. Discordance between these tests is rare.Empyema is a severe complication of pneumonia with high morbidity and mortality rates. Rapid diagnosis and tailoring of antibiotic therapy are crucial to treatment success for these severe bacterial lung infections. A Streptococcus pneumoniae . This case demonstrates discordance between the results of urinary and pleural fluid S. pneumoniae antigen tests, representing a potential pitfall in using rapid antigen testing on pleural fluid samples. False positives for the S. pneumoniae antigen in patients with viridans streptococci infections have been documented due to the cross-reactivity of cell wall proteins in different streptococcal species. Physicians encountering bacterial pneumonia of unknown etiology complicated by empyema should understand the potential for discordance and false positives using this diagnostic method.We report a case of a 69-year-old female with CT imaging findings consistent with empyema and a bronchopulmonary fistula. A rapid  Empyema, a severe complication of bacterial pneumonia, carries high rates of morbidity and mortality, where approximately 30% of patients require surgical intervention with drainage of the pleural space, and 15% of patients ultimately perish . IdentifRapid antigen tests offer the potential for fast and accurate diagnosis of common causative organisms implicated in empyema. In particular, Streptococcus pneumoniae (S. pneumoniae) rapid urine antigen tests demonstrate more sensitivity than blood cultures. Notably, using an S. pneumoniae antigen test drawn directly from the pleural fluid is equivalent to or enhances the diagnostic utility of the urinary antigen test alone ,8. DiscoThis case report was previously presented as a poster at the 2021 ACP Abstract Day Meeting at the University of Mississippi Medical Center on October 28th, 2021. Additionally, this case report was prepared using assistance from ChatGPT. However, no direct composition was provided by ChatGPT except where explicitly cited, and sources were reviewed and verified by the authors.A 69-year-old female with a past medical history of atrial fibrillation, hypertension, tobacco use, and anemia presented to the emergency department with a five-week history of malaise, cough, and progressive shortness of breath. The patient was admitted to the intensive care unit and treated for septic shock secondary to pneumonia. The patient entered atrial fibrillation with a rapid ventricular rate and began to vomit foul-smelling, purulent fluid. The patient quickly became unresponsive, necessitating intubation for emergent airway protection. A left chest tube was placed, returning tan-colored fluid identical to the patient's vomitus. CT imaging revealed findings consistent with empyema and necrotizing pneumonia with a bronchopulmonary fistula on axial Figure  and sagiIV vancomycin was started along with IV metronidazole and IV azithromycin. Pleural fluid, urine, blood, and bronchoscopy samples were taken for culture; Gram stain from pleural fluid reflected Gram-positive cocci in chains with Gram-negative rods. The patient was switched to IV vancomycin and IV piperacillin-tazobactam. Metronidazole and azithromycin were discontinued. Methicillin-resistant Staphylococcus aureus (MRSA), PCR, Legionella urine antigen, and Streptococcus urine antigen  resulted as negative; however, Streptococcus urine antigen test was positive when tested from a sample of pleural fluid taken from a chest tube directly. Pleural fluid cultures eventually grew as S. constellatus, a member of the viridans streptococci group, confirmed with VITEK 2 Compact 30 microbial identification system . The patient was transitioned to moxifloxacin by mouth daily with a plan to complete four to six weeks of therapy and transferred to a site with cardiothoracic surgery capability.This case demonstrates discordance between the results of urinary and pleural fluid rapid antigen tests and a false positive for S. pneumoniae, a potential pitfall in using rapid antigen assays on pleural fluid samples in empyema.Understanding the epidemiology of empyema is critical, as initial treatment is often performed empirically ,9. S. pnAs stated, rapid tests have been increasingly used as a fast, reliable answer to this clinical problem. The Binax-NOW rapid antigen test used in our case is an immunochromatographic test to detect S. pneumoniae membrane peptidoglycan C-polysaccharide in urine or cerebral spinal fluid . This raDiscordance between urinary and pleural fluid favors pleural fluid results when false positives occur. Patients often show a negative urinary antigen test and a positive pleural fluid antigen test confirmed by positive S. pneumoniae cultures ,7. In thPleural fluid rapid antigen testing represents a promising method to diagnose Streptococcus pneumoniae infection with a negative urinary antigen test or to confirm a positive urinary antigen test. However, physicians encountering bacterial pneumonia of unknown etiology complicated by empyema and utilizing this novel diagnostic method should understand the potential for discordance and false positives.


Infection rate of varicella zoster virus (VZV) is 95% in humans, and VZV infection is strongly associated with ischemic stroke (IS). However, the underlying molecular mechanisms of VZV-induced IS are still unclear, and there are no effective agents to treat and prevent VZV-induced IS.By integrating bioinformatics, this study explored the interactions between VZV and IS and potential medication to treat and prevent VZV-induced IS.In this study, the VZV and IS datasets from the GEO database were used to specify the common genes. Then, bioinformatics analysis including Gene Ontology, Kyoto Encyclopedia Genes Genomes and Protein–Protein Interaction network analysis was performed. Further, the hub genes, transcription factor (TF) gene interactions, TF-miRNA co-regulatory network and potential drug were obtained. Finally, validation was performed using molecular docking and molecular dynamics simulations.The potential molecular mechanisms of VZV-induced IS were studied using multiple bioinformatics tools. Ten hub genes were COL1A2, DCN, PDGFRB, ACTA2, etc. TF genes and miRNAs included JUN, FOS, CREB, BRCA1, PPARG, STAT3, miR-29, etc. A series of mechanism may be involved, such as inflammation, oxidative stress, blood–brain barrier disruption, foam cell generation and among others. Finally, we proposed resveratrol as a potential therapeutic medicine for the prevention and treatment of VZV-induced IS. Molecular docking and molecular dynamics results showed that resveratrol and hub genes exhibited strong binding score.Resveratrol could be an alternative for the prevention and treatment of VZV-IS. More in vivo and in vitro studies are needed in the future to fully explore the molecular mechanisms between VZV and IS and for medication development.The online version contains supplementary material available at 10.1186/s40001-023-01291-4. Stroke is the leading cause of death and disability, with nearly 100 million stroke patients worldwide . IS has Owing to infections, such as COVID-19, syphilis, and human immunodeficiency virus (HIV), the risk of IS have received more attention. However, the insidiousness and mild symptoms of VZV were often ignored, the most serious consequence of VZV was generally considered by the public to be postherpetic neuralgia (PHN) . Althoug−6 cm/s [Vaccines have been introduced in many countries to prevent VZV and provide good protection against VZV-induced IS. Studies have shown that VZV vaccines can reduce the incidence of VZV by up to 70% . Vaccina−6 cm/s . In addi−6 cm/s , 18.Our study sought to explore the underlying mechanism of VZV-induced IS. In this study, the datasets of VZV and IS were selected from the GEO database for analysis. First, we identified differentially expressed genes (DEGs) of VZV and IS, and the common DEGs served as the basis and raw data for the whole study. Further DEGs-based analyses, including Kyoto Encyclopedia Genes Genomes (KEGG) and Gene Ontology (GO) enrichment analysis, were performed to understand the biological processes of genome-based expression studies. Subsequently, protein–protein interaction (PPI) networks were used to identify hub genes from DEGs. Further, potential therapeutic agents were searched by hub genes. Finally, we performed molecular docking and molecular dynamics (MD) simulations of potential drugs and hub genes.All data were obtained from the GEO database. VZV data was from GSE175797, IS data was from GSE173719, GSE16561, and GSE22255. In addition, the Genecards [P < 0.05 for significantly differentially expressed genes. Owing to the small amount of sequencing data from VZV, we strictly screened the DEGs of IS to improve the accuracy of the study. Further, the Genecards database and the OMIM database were searched for "ischemic stroke" as a search term. To improve the accuracy of the obtained DEGs, we retained genes that were repeated twice as the IS targets. Finally, we compared the VZV and IS targets with the Human Protein Atlas database [DEGs for VZV were obtained from literature by Andrew N. Bubak et al. . The prodatabase  (proteinP < 0.05 is considered that GO and KEGG pathway was significantly enrichened [VZV-IS targets were imported into the Metascape website  (metascarichened .https://string-db.org/) was used to construct a PPI network with a confidence score ≥ 0.4. The hub genes of the PPI network were identified using the Cytoscape plugin cytohubba's degree algorithm and visualized using Cytoscape (v3.7.2) [STRING  (https:/(v3.7.2) . The top(v3.7.2) .NetworkAnalyst   for molecular docking. Potential hub genes for resveratrol are used for molecular docking. The 2D structure of resveratrol was downloaded from the PubChem database and crystal structures of core targets were downloaded from RCSB protein Data Bank (rcsb.org/) . Chem3D + and Cl− ions were added into the protein surface to neutralize the total charges of the systems. The resulting systems were solvated in a rectangular box of TIP3P waters extending up to minimum cutoff of 15 Å from the protein boundary [We performed MD simulations for 100 ns according to the conformation of the optimal binding score for molecular docking. The CHARMM36 force field was used to generate the parameters of the protein . Force fboundary . The Ambboundary . The sysboundary .Finally removing the unexpressed genes in the brain, we obtained a total of 1647 IS targets and 302 VZV targets. Using Venny, 63 overlap targets were obtained and considered as VZV-IS common targets , molecular function (MF) and cellular component (CC). These items were shown in Fig. KEGG analysis showed that these VZV-IS shared targets were enriched in focal adhesion, PI3K-Akt signaling pathway, ECM–receptor interaction, regulation of actin cytoskeleton, FoxO signaling pathway, adipocytokine signaling pathway, vascular smooth muscle contraction, estrogen signaling pathway, MAPK signaling pathway, cGMP-PKG signaling pathway, etc. Fig. .Fig. 4ToThe PPI network includes 48 nodes (four idle nodes are removed) and 192 edges, as shown in Fig. Ten hub genes were identified for TF genes Fig. . DCN is The TF-miRNA network was analyzed and provided a common interaction between miRNA and TF gene. This interaction may be involved in the regulation of hub gene expression. The TF-miRNA network consists of 166 nodes and 380 edges. 63 TF genes and 103 miRNAs formed the TF-miRNA network. The TF-miRNA co-regulatory network is shown in Fig. The specific information of 20 potential drugs for the treatment and prevention of VZV-IS is shown in Additional file By calculating binding score, the molecular docking results of resveratrol and targets’ proteins were predicted to be less than − 5.0 kcal/mol, indicating that these compounds had strong binding effect on key proteins. In other words, the lower the binding score of the ligand to the receptor, the more stable the binding conformation. It can be seen from Table To further study the interaction between resveratrol and the targets, we used MD to simulate the protein–resveratrol complex for 100 ns. The root-mean-square deviation (RMSD) from the average structure of backbone atoms for each MD trajectory was calculated as well for exploring the "position stability" for each complex. Figure VZV infection is a major risk factor for IS, especially in young adults without underlying disease , 42. PreThe ten hub genes are strongly associated with the occurrence and development of IS. (1) Heterozygous COL1A2 were found to be associated with IS . COL1A2 Furthermore, regulatory biomolecules are potential biomarkers for a variety of diseases, including IS. The miRNA and TF genes of hub gene were used to analyze the regulatory network, and a total of 103 miRNAs and 63 TF genes were identified. Among the TF genes with the strongest interaction, the degree values were JUN, FOS, CREB, BRCA1, PPARG, STAT3, JUND, SRF, FOXC1, Sox10, and ELK1. These TF genes also play an important role in IS. JUN plays an important role in neurodegeneration, cell death and repair after IS . CREB isWe further revealed more complex pathological processes based on GO and KEGG enrichment analysis of DEGs of VZV-IS and other findings. The GO and KEGG enrichment results can be mainly classified into the following categories: (1) inflammation and oxidative stress response, (2) regulation of vascular morphology and function, (3) regulation of extracellular matrix (ECM), (4) regulation of ion transport, and (5) regulation of cell adhesion. It is suggested that there are extremely complex regulatory mechanisms of VZV-induced IS. We have addressed these possible mechanisms.Inflammation and oxidative stress are the initial pathological processes after VZV activation. Multiple GO and KEGG items suggested that inflammation and oxidative stress played an important role in the process of VZV-IS, including antioxidant activity, oxidoreductase activity, hydrogen peroxide metabolic process, PI3K–Akt signaling pathway, FoxO signaling pathway, MAPK signaling pathway, cGMP–PKG signaling pathway, etc. VZV is first activated in the outer arterial membrane and early on there are neutrophils involved in the immune response, generating reactive oxygen species, leading to an imbalance in oxidative stress, inducing apoptosis, and loss of vascular smooth muscle cells –82. OverSubsequently, low-density lipoprotein (LDL) undergoes oxidation and accumulates in the vessel lining, and monocytes differentiate into macrophages that phagocytose oxidized LDL deposits and transform into foam cells, leading to the formation of atherosclerotic plaques . In the The ECM is an important component of the BBB structure. Immune thrombosis in COVID-19 is dominated by pulmonary venous and microvascular thrombosis –87. VZV Over time, vascular calcifications can occur in the intima or mesoderm of the arterial wall, increasing arterial stiffness . CalcifiKEGG and GO enrichment analysis are enriched in vascular smooth muscle contraction, vasculogenesis, blood vessel diameter maintenance, cGMP-PKG signaling pathway, and other items. This suggests that nitric oxide (NO) is also an important pathological process involved in VZV-IS. The suppression of VZV immunity that may result from NO production during inflammation is mediated by T cells . RegulatEstrogen pathway was found in KEGG enrichment of VZV-IS. We suggest that the estrogen protection against VZV in women is bidirectional. Studies have shown that the risk of VZV-induced atherosclerosis is higher in men than in women . 50.7% oGO enrichment results were enriched in calcium ion binding, regulation of calcium ion transport, regulation of calcium ion transport, and other items. It is suggested that the mechanism of VZV leading to atherosclerosis may be related to the regulation of calcium. Unlike the normal slow progression of atherosclerotic plaques, atherosclerosis due to infection is rapid and unstable. In particular, VZV directly invades the outer membrane of the cerebral arteries and then the inner membrane leading to atherosclerosis and stenosis within the cerebral arteries. In addition, due to the intense inflammation and immune response, this process is more rapid and intense than normal atherosclerotic plaque formation. Therefore, the year of VZV outbreak is a high-risk period for IS.Hyperlipidemia is one of the risk factors for VZV-induced IS. However, the study found that the lipid-lowering drug statin increased the risk of VZV development, a result that is clearly confusing , 99. ThiWe propose a possible mechanism by which the body's defense mechanisms are activated after VZV infection, inducing plaque to accumulate at the VZV-infected site, form plaques that wrap around the infected area, and prevent VZV transmission. The increased risk of VZV development by statin treatment may be due to that it is the body's defense mechanisms that induce the conversion of high-density lipoprotein cholesterol (HDL-C) to LDL, and statin treatment causes a lack of raw material for the formation of plaques covering the infected area, leading to an increased risk of plaque shedding. This also exposes a drawback of most drugs that they are too homogeneous in their therapeutic mechanisms, which is a fatal drawback in complex diseases. It suggests that we should adopt a multi-targeted therapy for the treatment and prevention of VZV-IS.The treatment of VZV-IS is now mainly based on the antiviral, hormonal, and symptomatic therapies , 103. ThWe screened 20 drugs based on the hub genes of VZV-IS. However, as a drug to protect or prevent VZV-induced IS, it should have several characteristics: (1) conform to Lipinski's rule of five, (2) have good bioavailability, (3) be able to cross the BBB, (4) have low toxicity, (5) be affordable, (6) be multitarget therapeutic, (7) have low adverse effects when used with other drugs, and (8) preferably already in wide use. Resveratrol, Melatonin, Menthol, and Aspirin conform all these characteristics.Resveratrol is the best choice among these four alternative drugs. Resveratrol is a natural product, widely available in grapes, easy to extract, and inexpensive . It can The recent studies have also found a protective effect of resveratrol in older females , with thMelatonin, a hormone secreted by the pineal gland in response to photoperiodic responses, has shown promising results in the treatment of several diseases, including IS and VZV , 130. ReIt is unclear whether the atherosclerotic changes result from a linear progression due to ongoing damage triggered by chronic infection with VZV or from periodic endothelial damage and incomplete healing due to reactivation of chronic infection to acutely activated infection. One view is that residual vascular inflammatory changes following an acute infection episode may persist and be further exacerbated by reactivation of new infection stimuli, and subsequent acute infections produce further damage to previously damaged and incompletely repaired vessel walls until this process develops into mature atherosclerotic lesions . HoweverIn terms of sequencing, there are no studies on VZV-IS. We have found the potential mechanism of VZV-IS by identifying DEGs between VZV and IS datasets. KEGG and GO enrichment results include regulation of inflammation and oxidative stress response, regulation of vascular morphology and function, regulation of extracellular matrix, regulation of ion transport, regulation of cell adhesion. Resveratrol is reasonably suggested as a drug because it is obtained by identifying hub genes, and it is a promising drug to become a therapeutic and preventive agent for VZV-IS. However, there is still no reliable animal model capable of simulating the VZV-IS pathological process. Simple animal models of VZV-infected middle cerebral artery occlusion do not representative of human VZV-IS pathological processes. Future development of stable animal models of VZV-IS is the basis for exploring the molecular mechanisms of VZV-IS and pharmacological experiments.Additional file 1. The specific information of 20 potential drugs for the treatment and prevention of VZV-IS.Additional file 2. Molecular docking information for VZV-IS drugs and their potential targets.


Campylobacter spp. is a gram-negative bacillus that causes infectious enteritis and consists of several species, including Campylobacter jejuni, Campylobacter coli, and Campylobacter fetus. Although C. jejuni and C. coli cause infectious enteritis primarily in immunocompetent hosts, C. fetus causes extraintestinal infections such as septicemia, meningitis, and perinatal infections in immunocompromised hosts, as well as myopericarditis in rare cases. Only a few cases of infectious myo(peri)carditis associated with C. coli in immunocompetent hosts have been reported. These studies concentrated on antecedent C. coli enterocolitis and never demonstrated a positive culture in the pericardial fluid.C. coli was identified from blood culture samples and blood-tinged pericardial fluid. He was successfully treated with antibacterial chemotherapy as well as pericardial fluid drainage and was discharged from the hospital with no complications.A 72-year-old Japanese man presented with a 2-week fever, cough, and vomiting lasting. He was on hemodialysis for polycystic kidney disease, as well as medication for diabetes and hypertension. A chest computed tomography (CT) scan and a transthoracic echocardiogram revealed bilateral pleural fluid and large pericardial fluid at the time of admission. C. coli in the pericardial fluid confirmed the diagnosis of C. coli pericarditis. C. coli may cause septic pericarditis in immunocompromised hosts, despite typically causing only enteritis.In this case, the presence of  Campylobacter spp. is well known as the causative agent of infectious enteritis. As of December 2014, Campylobacter had 26 species, two provisional species, and nine subspecies, including Campylobacter jejuni, Campylobacter coli, and Campylobacter fetus  of 178 patients) [C. coli was only found infrequently (16 [9%] of 178 patients) [C. coli is less prevalent than C. jejuni and C. fetus. There are few reports on C. coli-caused infectious diseases other than infectious enteritis.The bacteria emia  or pneumonia (33%) respectively [Salmonella and Shigella are uncommon but important gastrointestinal pathogens associated with myo(peri)carditis; however, the rising incidence of campylobacteriosis worldwide over the last decade has heightened interest in Campylobacter-associated myo(peri)carditis [C. fetus is almost always isolated from blood in cases where the pathogen has been linked to myopericarditis [C. fetus was isolated directly from pericardial fluid [C. coli-associated myo(peri)carditis has been described in a few case reports [C. coli enterocolitis, rather than C. coli growing from blood or pericardial fluid played a role. We present a case of infectious pericarditis caused by C. coli isolated from both pericardial fluid and blood in a patient who did not develop enterocolitis.Pericarditis is a pericardial inflammatory syndrome. Pericarditis and myocarditis have similar aetiologies, and overlapping forms are common in clinical practice. The majority of cases of acute pericarditis in developed countries are caused by viral infections or are autoreactive , and bacectively . Salmonecarditis . C. fetucarditis . In someneumonia 3% respecal fluid . C. coli reports , 11. TheKlebsiella pneumoniae, food poisoning caused by Salmonella spp. and oral candidiasis. He had no history of alcohol intake and was not infected with the human immunodeficiency virus.A 72-year-old Japanese man was admitted to the hospital with a fever and a large amount of pericardial fluid. He had been on hemodialysis for the past 19 years due to polycystic kidney disease. He had also been receiving treatment for hypertension and diabetes mellitus for the previous 32 years. His medical history included a urinary infection caused by He began coughing two weeks before his hospitalization. A chest X-ray and computed tomography (CT) scan one week before hospitalization revealed bilateral pleural fluid and pericardial fluid. An electrocardiogram taken three days before admission revealed flattening of the T wave and poor R wave progression Fig. . Hence, He was awake but vomited upon admission. There was no chest pain or enteritis in the patient. He had not eaten any raw or undercooked food prior to his admission. His physical exam revealed tachycardia, with a heart rate of 126 beats per minute, blood pressure of 172/84-mm Hg, and an axillary temperature of 38.6 °C. C-reactive protein , procalcitonin , hemoglobin A1c , and white blood cell  counts were all elevated. Creatinine phosphokinase , and lactate dehydrogenase  were, however, almost normal. Additional laboratory tests revealed that β-D-glucan levels were extremely high  and aspergillus antigen levels were mildly elevated . Anti-GPL-core IgA antibody, QuantiFERON TB-Gold plus, and cryptococcus antigen were all negative. A transthoracic echocardiogram revealed a significant amount of pericardial fluid. Spiral-shaped, gram-negative rods were cultured from admission blood (two sets) and on day 2 (two sets).C. coli or C. jejuni using Matrix-assisted laser desorption ionization time-of-flight analysis, but it could not tell the difference. Finally, using the Hippurate hydrolysis test, the organism was identified as C. coli. In the pericardial fluid culture, no mycobacteria or fungi grew.On day 2 of hospitalization, blood-tinged pericardial fluid was drained through pericardiocentesis. The pericardial fluid was examined and discovered to have pyogenic inflammation  as well as mildly elevated adenosine deaminase (ADA) levels (42.4 U/L). The pericardial pathological examination of the pericardial fluid revealed no bacterium, mycobacterium, fungus, or malignant findings. The spiral-shaped gram-negative rods grew in pericardial fluid culture, similar to the blood culture. Commercial automatic identification systems failed to recognize the organism. The organism was identified as C. coli bacteremic pericarditis was diagnosed.Drug susceptibility of the organism was evaluated. Minimum inhibitory concentrations against the organism were as follows: ampicillin, 1.0 mg/mL; imipenem, < 1.0 mg/mL; erythromycin, < 2.0 mg/mL; ciprofloxacin, < 0.25 mg/mL; and gentamicin, < 1.0 mg/mL. There stool was not microbiologically examined. Based on the findings, Meropenem  was administered intravenously for 44 days, followed by micafungin  for 15 days. On the sixth day of hospitalization, a transesophageal echocardiogram revealed no evidence of infective endocarditis. Furthermore, no organism grew from the blood on day 6 (two sets). He was discharged from the hospital on day 45 without any sequelae. His treatment was completed 14 days after he was discharged with oral clarithromycin . Even after 5 months after discharge, the patient never showed signs of disease relapse.Campylobacter-associated myo(peri)carditis [C. coli grew from blood and pericardial fluid. In the present case, it was unclear when, and where the patient became infected with C. coli in this case. He had no significant incidents, such as animal contact or eating raw meat. He is an elderly immunecompromised patient with diabetes and chronic renal failure on dialysis.Campylobacteriosis has become more common in the last decade, drawing attention to carditis . The obsCampylobacter causes myopericarditis is unknown, several intriguing putative mechanisms [C. fetus pericarditis appears to be the result of pericardium colonization following bacteremia in an immunocompromised host [C. fetus is almost always isolated from blood [C. fetus was isolated from blood in 90% (9/10) cases of C. fetus pericarditis. C. fetus myopericarditis could be caused by direct microbial invasion of the pericardium via the blood stream. On the other hand, C. jejuni causes myo(peri)carditis in immunocompetent hosts [C. jejuni was identified in 12 cases of C. jejuni-associated myo(peri)carditis through positive stool cultures and/or serological analyses, but it was never isolated from blood [C. jejun infection or a direct effect of Campylobacter spp. on the myocytes [C. coli has properties similar to C. jejuni and causes enterocolitis in immunocompetent people. C. coli-caused myo(peri)carditis in immunocompetent hosts with enteritis in the reported cases, and C. coli was identified from stool [C. coli caused pericarditis in an immunocompromised host who did not have enteritis, and C. coli was isolated from blood and pericardial fluid. Therefore, C. coli in the current case may directly invade the pericardium similar to C. fetus.Although the exact mechanism by which chanisms  have beesed host . When thom blood . C. fetuin 90% 9/ cases ofom stool . In the C. coli infections [C. coli is becoming increasingly resistant to them, according to recent reports [C. coli showed elevated minimum inhibitory concentrations to meropenem after long-term oral antibiotic treatment with tebipenem and faropenem for persistent infection, according to a case report [Quinolones and macrolides are frequently used in the treatment of fections . C. coli reports . In thise report . More reCandida or Aspergillus fungi were cultured from pericardial fluid, some cases of Candida pericarditis or aspergillus pericarditis were diagnosed [Laboratory testing revealed a highly elevated β-D-glucan (> 300 pg/mL) and a mildly elevated aspergillus antigen in this case (0.5 cutoff index). When β-D-glucan is high, it generally suggests the presence of fungal infection . When Caiagnosed –20. PhysThe pericardial fluid revealed a mildly elevated ADA (42.4 U/L) in the present case. High levels of ADA indicated tuberculous pericarditis. A recent meta-analysis of tuberculous pericarditis reported a pooled sensitivity of 0.90, specificity of 0.85, and negative possibility of 15% . HoweverC. coli-associated myo(peri)carditis is extremely rare; there have been a few reported cases without proof of positive culture in pericardial effusion. Because C. coli was isolated directly from the pericardial fluid in this case, the diagnosis was certain. C. coli has been suggested to cause septic pericarditis in immunocompromised hosts, although it typically causes enteritis.To summarize,


The correlation of adropin serum levels with mortality was better than MELD or Child-Pough scores . Higher adropin levels correlated with creatinine (r2 = 0.79. p < 0.01). Patients with diabetes mellitus and cardiovascular diseases had elevated adropin levels. Integrating adropin levels with the Child-Pugh and MELD scores improved their correlation with the time of death (correlation coefficient: 0.91 vs. 0.38 and 0.67 vs. 0.32). The data of this feasibility study suggest that combining serum adropin with the Child-Pugh score and MELD-Na score improves the prediction of mortality in cirrhosis and can serve as a measure for assessing kidney dysfunction in these patientsAdropin is a peptide that was suggested to have a role in cirrhosis. The present study aimed to determine the ability to use serum adropin levels to improve their prediction accuracy as an adjunct to the current scores. In a single-center, proof-of-concept study, serum adropin levels were determined in thirty-three cirrhotic patients. The data were analyzed in correlation with Child-Pugh and MELD-Na scores, laboratory parameters, and mortality. Adropin levels were higher among cirrhotic patients that died within 180 days (1,325.7 ng/dL vs. 870.3 ng/dL,   Cirrhosis is a progressive fibrosing nodular condition that disrupts the entire typical architecture of the liver . More thThe Child-Pugh score (CPS) classification and the Model for end-stage liver disease (MELD) score are commonly used models for predicting mortality in cirrhosis . HoweverAdropin is a 76-amino-acids-secreted peptide encoded by the Enho gene and is conserved among humans, mice, and rats . The phyThe present single-center proof-of-concept study aimed to determine the potential of using serum levels of adropin as a prognostic biomarker in patients with chronic liver disease and to determine its use as an adjunct to MELD and CPS for improving their performance in predicting mortality.This single-center prospective, observational study was approved by the Institutional Review Board (IRB) . Participants signed informed consent during enrollment as defined by the local IRB.Adult subjects (18–80 years) with chronic liver disease of all etiologies were enrolled. The main exclusion criteria were evidence of other acute severe disease or any acute medical condition within 48 h of blood tests. Controls were adults without known liver disease.A single blood test for serum levels of adropin was obtained. Serum was collected using a serum separator tube and centrifuged for 20 min at 1,000 × g. Samples were stored in aliquots at −80°C. Adropin levels were determined using a sandwich enzyme immunoassay kit for adropin  according to the manufacturer’s procedure. The optical absorbance was measured spectrophotometrically at 450 nm.Subjects’ clinical and laboratory data were generated from the patient’s medical records.p-value of 0.05 or less was considered statistically significant.Comparing adropin serum levels between cirrhotic and non-cirrhotic patients was carried out using the non-parametric Mann-Whitney test. The same test was used to compare different scoring systems trying to predict mortality. ROC analysis was used when the score was significantly different between groups. The Kruskal-Wallis non-parametric test was applied to compare Adropin levels between three independent groups. The non-parametric tests were used due to the small sample size and the non-normal distribution of adropin levels in some subgroups compared. The Pearson correlation coefficient was calculated to assess the strength of the linear association between adropin levels and other quantitative variables. All tests applied were two-tailed, and a Clinical and laboratory data were obtained from 33 cirrhotic patients . Sixteenp = 0.39). Hepatocellular carcinoma (HCC) was diagnosed in 27.3%. Five-of the HCC patients had NASH, and four with viral hepatitis.The average MELD-sodium (MELD-Na) score was 16.0, corresponding with most patients (60.6%) in Child-Pough group B. The severity was marginally higher in patients with NASH cirrhosis, albeit not reaching significance  and higher creatinine levels .The laboratory values are described in p = 0.37).Serum adropin levels were higher among patients with chronic liver disease relative to controls, albeit not reaching statistical significance . Patients with chronic liver diseases not attributed to the above causes had non-significantly higher adropin levels . It included patients diagnosed with biliary cirrhosis, DILI, cardiac cirrhosis, sickle cell disease, and cryptogenic cirrhosis. Finally, adropin levels were not affected by the development of HCC .Among the patients with chronic liver disease, subjects with NASH and viral-mediated liver disease had similar serum levels of adropin, 973.1 ng/dL and 946.4 ng/dL, respectively. Lower serum Adropin levels were noted among patients with autoimmune hepatitis relative to all other diagnoses , and including the healthy controls, the sex difference widened .Adropin levels were higher in males with chronic liver disease . The glomerular filtration rate (GFR) was calculated according to the 2021 CKD-EPI equation. Calculated GFR also correlated with adropin levels .A negative correlation trend was noted with alpha-fetoprotein . A similar effect was noted among patients with cardiovascular diseases . No effect was noted in patients with hypertension and hyperlipidemia . The area under the curve (AUC) in ROC analysis for adropin to predict mortality within 180 days was 0.76. When implementing adropin levels of 1,058 ng/dl as a cut-off point, sensitivity and specificity for death during the following 180 days were 83% and 80%, respectively. MELD-Na, but not CPS, was also significantly higher in patients who died 6 months after being tested  with an AUC of 0.8. An integration of adropin levels with the CPS (p = 0.006), with an AUC of 0.8 (Eight patients (24.2%) died between 7 and 233 days following enrollment in the study. Adropin levels were higher among the six patients that died within 180 days  . An integration of adropin levels with the CPS (r2 = 0.91).Adropin levels were inversely correlated to the time until death ( = 0.74) . Adropinp = 0.082). Correlation with the time of death was not as precise (r2 = 0.67) as the combination with CPS (An integration of Adropin levels and MELD-Na (with CPS .This feasibility single-center trial showed that serum levels of Adropin predicted mortality better than the often-used MELD-Na and CPS. Moreover, adding Adropin levels to the Child-Pugh and MELD-Na scores improved mortality prediction. Serum levels of adropin levels positively correlated with poor prognostic factors such as mortality and kidney injury.Serum adropin levels decreased and negatively correlated with liver injury in NASH mice. Knockout of Adropin significantly exacerbated hepatic steatosis, inflammatory responses, and fibrosis in mice. Administration of Adropin bioactive peptides slowed NASH progression in mice . In humaPrevious studies demonstrated that higher adropin levels correlated with a better metabolic profile , 21—the The correlation between adropin and creatinine was significant. Previous studies regarding adropin and kidney function focused mainly on diabetic nephropathy and showed a negative correlation between adropin and nephropathy progression , 23. HepThis study is limited by the relatively small number of patients and being a single-center study. The significance of the data supports extensive studies for determining the potential role of adropin as a biomarker and therapeutic target in these patients. In addition, the exact cause of death was unknown, preventing us from inferring about adropin relation to cirrhosis-related deaths. However, as cirrhosis is a significant driver of morbidity and mortality, an association with all-cause mortality is essential for medical decision-making.In summary, elevated serum adropin levels are a poor prognostic factor in patients with chronic liver disease independent of the etiology. Adropin was superior to the standard prognostic models, CPS and MELD-Na, in predicting mortality and correlated with decreased renal function in cirrhotic patients. It can serve as a variable for improving the prediction performance of current scores. Larger cohorts are expected to shed light on the potential use of adropin as an additional biomarker to diagnose better and predict the prognosis of chronic liver disease and as a potential new therapeutic target in cirrhosis and hepatorenal syndrome.


Pectoralis major ultimate pH, a proxy for muscle glycogen stores, characterization of the yolk and amniotic fluid revealed a different nutritional environment. The present study aimed to assess indicators of embryo metabolism in pHu lines (pHu+ and pHu−) using allantoic fluids (compartment storing nitrogenous waste products and metabolites), collected at days 10, 14 and 17 of embryogenesis and characterized by 1H-NMR spectroscopy. Analysis of metabolic profiles revealed a significant stage effect, with an enrichment in metabolites at the end of incubation, and an increase in interindividual variability during development. OPLS-DA analysis discriminated the two lines. The allantoic fluid of pHu− was richer in carbohydrates, intermediates of purine metabolism and derivatives of tryptophan-histidine metabolism, while formate, branched-chain amino acids, Krebs cycle intermediates and metabolites from different catabolic pathways were more abundant in pHu+. In conclusion, the characterization of the main nutrient sources for embryos and now allantoic fluids provided an overview of the in ovo nutritional environment of pHu lines. Moreover, this study revealed the establishment, as early as day 10 of embryo development, of specific metabolic signatures in the allantoic fluid of pHu+ and pHu− lines.Nutrient availability in eggs can affect early metabolic orientation in birds. In chickens divergently selected on the  Access to O2 fully supports the complete combustion of fatty acids, which are used as the primary source of energy for embryo development2. According to Nasir and Peebles3, intensive selection for a high growth rate and muscle yield has changed the nutrient requirements of chicken embryos, with a potential imbalance between nutrient requirements and reserves. pHu+ and pHu− lines have been divergently selected based on the ultimate pH (pHu) of the meat, a proxy for muscle glycogen reserves4. The effects of this selection on broiler metabolism have been studied at 6 weeks of age through the quantification of muscle and serum metabolites by nuclear magnetic resonance (NMR) spectroscopy5. Chickens with low muscle pHu (pHu−) and high glycogen reserves are characterized by an over-representation of carbohydrate-related metabolites in serum and muscle, while chickens with high muscle pHu (pHu +) and low glycogen reserves exhibit markers of oxidative stress, muscle proteolysis, amino acid catabolism and β-oxidation. A liquid chromatography–mass spectrometry approach performed on 17-day-old animals also revealed different blood lipidomic signatures between the two divergent lines6. Glycerolipids (involved in energy storage) and ceramides were in higher concentrations in the blood of pHu− birds, whereas glycerophospholipids and sterols were more abundant in pHu+. Finally, it has been shown that differences between lines related to protein and energy metabolism are established from hatching7.During the incubation period (21 days in chickens), the embryo depends essentially on the nutrient content of the egg and its metabolism shifts in accordance with the type of substrate available and oxygen supplyin ovo nutrient availability, the metabolome of the yolk and amniotic fluid was studied in pHu− and pHu+ lines before incubation (E0) and at day 10 of embryo development (E10)8. Metabolomic analysis of these two compartments revealed differences in metabolite composition and abundance in embryonated eggs of pHu+ and pHu− lines, which may explain subsequent metabolic and developmental specificities. To further describe the nutritional environment of the embryos, the present study investigated the metabolic composition of the allantoic fluid of both lines during development. Indeed, the allantoic fluid can store nitrogenous waste products, free amino acids and other important compounds for late embryo nutrition11.To understand the metabolic orientation of embryos and the potential link with 13. According to Da silva et al.10, some nutrients, such as proteins and peptides, secreted into the allantoic fluid can be digested to provide free amino acids, which can be reabsorbed by the allantoic membrane and redirected to the embryo via the chorioallantoic capillary plexus. Since the allantoic fluid pH decreases in the last stages of development to reach intestinal pH values, optimal conditions can be found in the allantois for protein pre-digestion (activation of proteases).Allantois of the chick embryo appears at about 3 days of incubation as an evagination of the gut wall. Soon after its appearance, a branched network of capillaries is formed. It receives blood via the allantoic arteries and returns it to the body via the allantoic veins. Several studies revealed that the blood, allantoic and amniotic fluid of chicken embryos, which are separated from each other by complex and specific barriers, contain many amino acids and related compounds1H-NMR spectroscopy was conducted on allantoic fluids at E10, E14 and E17. These stages were chosen according to the availability of substrates in the eggs and the literature on embryo development and metabolism16. At E10, yolk lipids are used as the main energy substrate. The stage E14 is characterized by the complete differentiation of the chorioallantoic membrane, an intensification of nutrient transfers to the embryonic tissues , the opening of the sero-amniotic connection allowing the albumen to enter the amniotic cavity and by a rapid growth of the embryos. At E17, nutrient sources diversify, the growth rate decreases and the embryo prepares to hatch. The objective was to obtain indirect indicators from the metabolic status of the embryos and to determine the stage at which metabolic orientation was established in the two divergent lines.To assess the developmental patterns of pHu+ and pHu− metabolism during embryogenesis, a metabolomic analysis by 1H-NMR spectroscopy at E10, E14 and E17. Figure 8. Forty distinct metabolites were identified in the allantoic fluid of the pHu lines  and clearly discriminated the pHu+ and pHu− lines. The reliability of this model was assessed by CV-ANOVA (P = 5.1e-05) and the predictive ability Q2 was 0.81. The importance of the metabolites in the model and their contribution are presented in Fig. At E10, the model adjusted from the metabolome of both lines was composed of 1 predictive and 4 orthogonal components Fig. A. It incAmong the 16 discriminating metabolites, we identified amino acids , organic compounds (dimethylamine), vitamins , sugars (mannose), Krebs cycle intermediates (fumarate), metabolites related to purines and pyrimidines  and muscle metabolism (creatine). The 3-HOisobutyrate, resulting from the partial degradation of branched chain amino acids , was also discriminant between both lines. Choline, creatine, glycine, 3-HOisobutyrate, hypoxanthine and nicotinate were more abundant in the allantoic fluid of pHu−, while cytidine, dimethylamine, fumarate, histidine, isoleucine, leucine, mannose, tyrosine and valine were more abundant in the pHu+ line.2Y) and clearly discriminated the pHu+ and pHu− lines. The reliability of this model was assessed by CV-ANOVA (P = 0.00027), and the predictive ability (Q2) was 0.56. The importance of the metabolites in the model and their contribution are presented in Fig. At E14, the model adjusted from the metabolome of both lines was composed of 1 predictive and 1 orthogonal component , organic compounds (dimethylsulfone), metabolites related to purines and pyrimidines , and metabolites involved in one-carbon metabolism . Most of them  were overrepresented in the allantoic fluid of pHu−. Only formate was more abundant in the pHu+ line.2Y) and clearly discriminated the pHu+ and pHu− lines. The reliability of this model was assessed by CV-ANOVA (P = 0.0022), and the predictive ability (Q2) was 0.55. The importance of the metabolites in the model and their contribution are presented in Fig. At E17, the model adjusted from the metabolome of both lines was composed of 1 predictive and 1 orthogonal component Fig. A. It incAmong the 25 discriminating metabolites, we identified amino acids and derivatives , metabolites related to purines and pyrimidines , organic compounds , sugars (glucose), Krebs cycle intermediates  and formate, a one-carbon metabolism intermediate. Betaine was considered a methyl group donor and organic osmolyte, but O-phosphocholine, an intermediate in the biosynthesis of phosphatidylcholine, was also detected. Glucose, dimethylamine, isoleucine, kynurenine, O-phosphocholine, phenylalanine and urocanate were overrepresented in the allantoic fluid of pHu−, while 3-HObutyrate, 3-HOisobutyrate, 3-methyl-2-oxovalerate, alanine, betaine, citrate, cytidine, dimethylsulfone, formate, fumarate, inosine, lactate, lysine and uridine-cytidine were overrepresented in the pHu+ line.1H-NMR spectroscopy, its quantitative accuracy and its ability to identify structures17, this instrumental approach was of great interest to characterize the metabolome of different egg compartments such as yolk, amniotic fluid8 and allantoic fluid.Because of the excellent reproducibility of 18. Our results are in agreement with the increase in amino acid concentration in allantoic fluid from E13 to E18 observed by Zhu et al.19. Changes in relative metabolic contents during in ovo development  reflect significant changes in embryo metabolism, transfers of compounds between egg compartments13 or activation of proteases in relation to environmental acidification10. More generally, the accumulation mentioned above was particularly strong for some metabolites, with a fold change greater than 5. The classes mainly concerned with this high accumulation were amino acids, peptides and analogues, carbonyl compounds, and those related to purine metabolism .In birds, the composition of allantoic fluid and its evolution during embryonic development are still poorly described. The OPLS-DA performed on the spectral dataset clearly identified three clusters highlighting differences in the metabolic profiles obtained at E10, E14 and E17, regardless of the line. It also revealed an increase in inter-individual variability during embryonic development. With the exception of formate, nicotinate and phenylalanine, all metabolites accumulated in the allantoic fluid, especially amino acids, peptides and analogues, the most represented class among those identified with 12 distinct metabolites . In addition to serving as building blocks for tissue protein synthesis, amino acids act as antioxidants, regulators of hormone secretion, major fuels for embryonic growth and cell signaling molecules. Amino acids are also essential precursors for the synthesis of non-protein substances with biological importance, including purine, pyrimidine nucleotides and creatine8. Among the 40 metabolites identified in the allantoic fluid, 31 were present in the yolk and/or amniotic fluid. The 9 metabolites specifically identified in the allantoic fluid include several compounds involved in tryptophan metabolism , amino acid catabolism , Krebs cycle (cis-aconitate), and purine and pyrimidine metabolism (xanthosine and uracil).Characterization of allantoic fluid in addition to yolk and amniotic fluid provided an overview of the nutritional environment of pHu+ and pHu− embryosPectoralis major muscle were strongly discriminant and revealed line-related differences in both energy metabolism and one-carbon and purine metabolism.Regardless of the stage , the OPLS-DA models based on the metabolome of the allantoic fluid of the two genetic lines divergently selected for high (pHu +) or low (pHu−) ultimate pH of the 8. Leucine was more abundant in the egg yolk of pHu+ at E0, while leucine and isoleucine were more abundant in the amniotic fluid at E10. Valine was specifically discriminant in the allantoic fluid and only at E10.At E10, the allantoic fluid of the pHu+ contained more branched amino acids (BCAAs), such as leucine, isoleucine and valine, than that of pHu−, which was also the case for some of them in the yolk and amniotic fluid20, the Krebs cycle is responsible for the complete oxidation of acetyl-CoA and the formation of intermediates required for adenosine triphosphate (ATP) production and other anabolic pathways. An overrepresentation of Krebs cycle intermediates, such as fumarate and citrate, was also observed in the allantoic fluid of the pHu+ line at E17. These results and the high concentration of alanine (the main glucogenic agent) in the pHu+ suggest a greater catabolism of amino acids in this line to produce energy, probably favored by the greater availability of BCAAs in the different extraembryonic structures . The higher abundance of 3-methyl-2-oxovalerate, 3-HObutyrate and betaine might also indicate a higher use of lipids as energy substrates in pHu+ compared to pHu− embryos. According to Zhang et al.21, betaine is involved in lipid metabolism by playing a role in the inhibition of fatty acid synthesis and in the stimulation of fatty acid oxidation and lipid secretion. Furthermore, 3-HObutyrate is known to act as a signal to regulate metabolism and maintain energy homeostasis during nutrient deprivation22. Overall, the differences observed in allantoic fluid at E17 are consistent with previous metabolomic studies performed at 6 weeks of age in serum and Pectoralis major muscle of pHu+ and pHu− lines, which showed that pHu+ chickens have a greater resort to lipid and amino acid catabolism, likely due to their low level of energy substrates compared to pHu−5.These differences disappeared at E14 but later in development , the allantoic fluid of the pHu+ contained more 3-HObutyrate, 3-methyl-2-oxovalerate and 3-HOisobutyrate. Metabolites resulting from the catabolism of BCAAs are converted into acetyl-CoA, which acts mainly as a substrate for the Krebs cycle. As specified by Hada et al.8. Creatine is deposited by the hen in the yolk and albumen23 but is also synthesized de novo by the embryo24. It is a key compound that plays an important role in energy metabolism25. According to Muccini et al.26, creatine can maintain ATP turnover, acid–base balance and mitochondrial function. However, creatine synthesis considerably solicits arginine and methionine metabolism27. Approximately 50% of all S-adenosylmethionine (SAM)-derived methyl groups are consumed by guanidinoacetate methylation, the final step in creatine synthesis28. At E17, the allantoic fluid of pHu− contained more glucose, suggesting a greater availability of carbohydrates in the allantoic fluid of pHu− as hatching (a very energy-consuming process) approaches. The fact that the end product of anaerobic glycolysis, lactate, is less abundant in pHu− than pHu+ at this stage may indicate that aerobic recycling of lactate is less efficient in pHu+.Compared to pHu+, creatine was found in higher abundance in the allantoic fluid of pHu− at E10, similar to what was observed in amniotic fluid at the same stage29. In humans, nicotinate stimulates muscle mitochondrial biogenesis and respiratory chain activity30. Due to its anti-inflammatory, anti-oxidant and anti-apoptotic activities in a variety of cells and tissues, nicotinate is actively involved in the prevention of many pathological processes32. Nicotinate is mainly synthesized in the liver through the kynurenine pathway, which is the major catabolic route of tryptophan34. From E14 onwards, nicotinate was not detected in the allantoic fluid, while kynurenine accumulated, and to a greater extent, in the pHu− line compared to the pHu+ line.Nicotinate, also known as niacin or vitamin B3, was found in greater abundance in the allantoic fluid of pHu− at E10. Nicotinate is a water-soluble vitamin that plays a central role in energy metabolism and oxidative phosphorylation. In particular, it is a precursor of the bioactive molecules nicotinamide adenine dinucleotide (NAD) and nicotinamide adenine dinucleotide phosphate (NADP)8. The present study showed no difference at E10 but a higher abundance of formate in pHu+ allantoic fluid at E14 and E17 compared to pHu−. At E17, betaine was also more abundant in the allantoic fluid of pHu+. Formate and betaine are both involved in one-carbon metabolism, which includes the folate and methionine cycles, as well as the trans-sulfuration pathway , respectively. This may suggest a greater utilization of this metabolite in the last third of in ovo development, due to greater requirements for growth and metabolism in the pHu+ line.Formate is produced via quite a number of processes, from different substrates, in different tissues, and in different cellular organelles. These metabolic processes occur through both folate-independent and folate-dependent reactions Fig. . RegardiSeveral metabolites involved in purine metabolism also discriminated against the two lines. Among them, xanthosine and hypoxanthine  were more abundant in the allantoic fluid of the pHu− line until E14, which could explain the higher uric acid content in this line at E10 . In contrast, cytidine and inosine were more abundant in the pHu+ line at E17. Purine degradation seems to be more important in the pHu− line than in the pHu+ line, which is also characterized by a higher abundance of nucleosides (nucleic base + ribose). As de novo synthesis is very energy intensive, nucleosides could be reused through the purine recycling or rescue pathway, as evidenced by the higher abundance of inosine (hypoxanthine + ribose).40. However, in our study, it is likely that betaine is considered more of an osmolyte, a molecule required for the maintenance of cellular integrity 41. Indeed, a recent study showed that eggshell percentage, as well as eggshell thickness, were lower in pHu+, suggesting greater gas exchange and water loss in this line42. At E17, the higher betaine content in the allantoic fluid of pHu+ could therefore be a sign of a stronger adaptive response to ensure embryo protection. Other metabolites involved in embryo protection, such as glycine and glutamate, however, were more abundant in the pHu− line. These amino acids are involved in the production of glutathione, a major contributor to the redox balance in cells, through its ability to scavenge and reduce reactive oxygen species (ROS)43.Betaine, presented above as a marker of fatty acid oxidation in the pHu+ line, is also considered a methyl group donorIn conclusion, the characterization of allantoic fluid by a metabolomics approach revealed different metabolic profiles during embryogenesis, as well as between the two divergent lines for their muscle glycogen reserves. At E10, pHu+ and pHu− lines have already shown a specific metabolic signature. Due to the lower availability of energy substrates, such as glucose and creatine, pHu+ embryos may use different catabolic pathways, such as amino acid catabolism and lipid oxidation, to produce energy. Finally, our study revealed that allantoic fluid could provide early and potentially accessible biomarkers of embryo metabolism.https://doi.org/10.15454/1.5572326250887292E12) , as described in Petit et al.8. The study was conducted on eggs issued from the tenth generation of the two genetic lines divergently selected for high or low breast meat ultimate pH . Fertile eggs were obtained from laying hens (pHu+ and pHu−) at the peak of laying (28–29 weeks of age). Eggs were incubated under standard conditions at 37.8 °C and 56% relative humidity in the experimental hatchery of PEAT to collect the allantoic fluid on days 10 (E10), 14 (E14) and 17 (E17) of incubation. These samples were collected from the same experimental design as the yolk and amniotic fluids analyzed previously8. As a result, the eggs were different at each stage of sampling and egg incubation was stopped.All investigators were certified by the French government to handle animal experiments. The experiments were carried out at the PEAT INRAE Poultry Experimental Facility  using a syringe. The samples were centrifuged to remove cellular debris . The supernatants were ultra-filtered on Amicon columns (cut-off 3 kDa) to remove proteins and peptides of high molecular weight. The ultrafiltrates were stored at − 80 °C until NMR analysis. Ultrafiltrate samples (150 µL) were prepared by the addition of 50 µL of phosphate buffer prepared in deuterium oxide  and 10 µL of trimethylsilylpropionic acid , used as an internal reference in NMR spectroscopy. Quality controls were prepared to assess analytical reproducibility by pooling 80 µL of randomly selected samples.1H-NMR spectra were acquired on a Bruker DRX-600 Avance III HD spectrometer  equipped with a TCI cryoprobe using a “noesypr1d” pulse sequence with a relaxation delay of 20 s, a time domain of 64 K data points, and a spectral width of 12 ppm and 64 scans. The spectra were phased, the baseline corrected and the chemical shifts referenced to the TSP. The spectra were reduced into consecutive and non-overlapping spectral regions (buckets) that were integrated and normalized by the TSP. The integration of those buckets was proportional to the concentration of the metabolites but also to the number of protons that resonate at the frequency of the peaks. Thus, each bucket represented a variable. This allowed the generation of a data matrix that was analyzed by multivariate statistics. Peak assignments were done using databases such as the Human Metabolome Database (http://www.hmdb.ca) and Chenomx NMR suite 8.1 evaluation edition .The 2Y) and the predictive ability of the model (Q2). Variance analysis (CV-ANOVA) was then applied to further evaluate the significance of the results5. The OPLS-DA were interpreted through their score plots—a projection of the linear combinations of the metabolomics data representative of each sample on two components—and their loading plots. The loading plots represent the projection of the correlation structure of the metabolic variables (X), and the stages and lines variables (Y) on the axis pq1 et pq2 which are the combination in one vector of the loading of X weighted for the importance of the metabolites in the approximation of X (p) and the loading of Y weighted by the importance of the classes in the approximation of Y variation correlated to X (q) in the component 1 (pq1) or 2 (pq2).Principal component analysis (PCA) followed by orthogonal partial least squares’ discriminant analysis (OPLS-DA) was performed using the SIMCA 16 software  on the dataset. All data were scaled to unit variance. OPLS-DA is a supervised approach to discriminate groups, i.e., embryonic stage and line, using metabolomic data. Samples identified as being out of the confidence interval in the PCA (outliers) were removed from the analysis. For optimal model classification, variables with low regression coefficients and wide confidence intervals combined with low variable importance in projection (VIP) were iteratively removed. The overall quality of the models was appreciated by the proportion of variance in Y explained by the predictive component of the model (RIn Tables https://arriveguidelines.org/) have been respected. All investigators were certified by the French government to handle animal experiments.All animal care and experimental procedures needed for this study were conducted in accordance with current European legislation (EU Directive 2010/63/EU) and were approved by the Ethics Committee for Animal Experimentation of Val de Loire (CEEA VdL). This ethics committee is registered by the National Committee under the number C2EA 19. The use of all our live animals is respectful of animal welfare and has been thought through according to the 3Rs rule . The ARRIVE guidelines were followed while reporting the experiments on animals. The points of the guide (https://zenodo.org; https://doi.org/10.5281/zenodo.7736015.All data and spectra generated or analyzed during this study are included in the supplementary data file and have been deposited on Supplementary Information 1.Supplementary Information 2.


Proc. R. Soc. B283, 20152694. (Published online 25 May 2016). (https://dx.doi.org/10.1098/rspb.2015.2694)Proceedings B is committed to the integrity of the scientific record. It therefore takes allegations concerning the validity of research published in the journal extremely seriously. At same time, however, it is conscious of the potentially far-reaching consequences of these allegations for authors and for the institutions in which they work. We therefore aim to take a fair and balanced approach to investigations on the reliability of research by thoroughly assessing the combined weight of evidence that is made available to us at any given time when deciding if an article requires a correction or a retraction. Although assessments may be informed by investigations by other journals and/or institutions, ultimately Proceedings B only investigates and adjudicates articles that it has published.Proceedings B was first made aware of concerns regarding an article by Scott & Dixson published in 2016 [When Subsequently, a new set of allegations surfaced; some submitted to the journal and others made in the public domain. The team of editors was tasked with further assessment of the combined weight of this body of evidence, including reports from two external experts on flume experiments. We conducted a detailed quantitative assessment of the Scott & Dixson data as well as analyses provided to us by several whistle blowers. Our conclusion at that time was that there was insufficient evidence to warrant retraction . AlthougProceedings B has now lost confidence in the reliability of the data that provide the basis for the main findings and conclusions in Scott and Dixson (2016) and has therefore decided to retract the article and the associated correction.Recently, Southern Cross University, the employer of the first and corresponding author Dr Anna Scott advised us of substantial doubts regarding the new timelines stated in the correction and the feasibility of the experimental design. They requested that we retract the Scott and Dixson (2016) article as well as its correction. Therefore, having considered the full body of evidence, including these new disclosures,


This investigation shows the effect of slippage on the slow spinning of a rigid sphere covered by a non-concentric spherical hollow full of an incompressible couple stress fluid. Moreover, the velocity slip conditions are employed on surfaces of both the rigid sphere and the cavity. In addition, the solid sphere and the cavity are rotating axially at various angular speeds. The solution is obtained semi-analytically at low Reynolds numbers utilizing the superposition with the numerical collocation approach. This paper discusses the hydrodynamic couple exerted by the fluid on the internal particle. The dimensionless torque increases as the slip and spin slip increase by 99%, the couple stress parameter by 49%, and the separation parameter by 79%. Additionally, the non-dimensional torque decreases with the increase of the size ratio by 89%. Consequently, it is found that all the results agreed with the corresponding numerical analysis in the traditional viscous liquids and the revolving of two eccentric rigid spheres with no slippage :1–9, 2022). Couple stress fluids are fluids that are suspended in a viscous medium with stiff particles that are randomly oriented. This theory, which is the most direct popularization of the theory of the traditional fluid, was created by Stokes. Moreover, a couple of conflicts and physical ties are maintained. The theory of couple stress fluid is thoroughly outlined in the work by Stokes2. Numerous studies into a few stress fluids, such as5. Recently, Al-Hanaya et al.6 examined the axisymmetric motion of fluid couple stress between two eccentric spinning spheres that the normalized torque on the solid sphere rotates inside the outside sphere with no slippages. Moreover, the normalized torque increases with both the size ratio and separation distance. The couple stress fluids have been applied in several applications of porous mediums such as El-Sapa and Almoneef7 investigated the axisymmetric movement of an aerosol particle contained in a couple-stress fluid moving in a slippage regime. On the other hand, Maurya et al.8 evaluated the couple stress fluid flow surrounding a solid sphere in a porous material in the presence of a homogeneous magnetic field.Recent research on the theory of couple stress fluid behavior is very helpful, and these analyses explain enough about the habits of rheological compound liquids like suspensions of polymers with molecules from prolonged chains, lubricants, liquid crystals, and blood. Therefore, the theory of couple stress fluid, which Stokes first proposed in 196610. Additionally, Ellahi11 examined how the slip boundary condition affected non-Newtonian flows in a channel. He used the symmetric analysis method (HAM) to solve nonlinear boundary value governing problems. The flows of Couette, Poiseuille, and modified Couette of an incompressible even-pressure flow between parallel walls were all correctly solved by Devakar et al.12 using slip limit conditions. The solution is like a viscous classical Newtonian fluid solution with pair pressures close to zero in the finite state. Additionally, the results demonstrate that different pressures reduce fluid velocity. A rigid sliding ball's creeping motion in an infinite binary stress fluid was studied by Ashmawy13. He also added the vanishing couple stress condition and the linear sliding limit condition to the sphere's surface. Thus, Slippage conditions at the boundary are used to study the unstable flow of an incompressible couple stress fluid between two plates that are in parallel. by Saad and Ashmawy14. Further, the two rigid spheres with slippage surfaces that have distinct diameters and are suspended in a porous medium in a horizontal magnetic field were studied semi-analytically for the Stokes flow approximation15.In the study of fluid dynamics, employing the traditional no-slip boundaries condition is prevalent. Consequently, the no-slip requirement, however, may not always hold, and fluid particle slippage on the surface of the stiff barrier can occur, according to various investigations conducted in the past century16. The Stokes flow spinning of a spheroid around its axis of revolutions in a viscous fluid with the slipping influence was further studied by Chang and Keh17 where the torque monotonically decreases with the slippage parameter for a spheroid with a constant size ratio. On the other side, at low Reynolds numbers, Lee and Keh studied the static rotation of a sliding spherical particle about the longitudinal axis of a sliding circular tube18. The creeping flow of continuous spinning of a slipping sphere that is only slightly distorted in a Brinkman medium was investigated analytically in19.Medical and industrial applications require porous material fluid flow quantification. Chemical, biological, and environmental engineering and research gently spin a solid particle in an incompressible porous medium, non-Newtonian, or Newtonian fluids. Furthermore, Jeffrey solved limitless Newtonian fluid spheroid rotation20. Therefore, it is important to understand if particle rotation is impacted by the presence of boundary walls. So, the slow rotations of a hard sphere near one or more planar walls21, inside a spherical hole22 were studied. Also, theoretical research has investigated the rotary motion of a soft or porous circular object with a low Reynolds number in a cylinder23 and a spherical cavity25. These investigations demonstrate that borders can have a significant and fascinating impact on how particles rotate. In addition, numerous researchers used the collocation method to solve various fluid dynamics problems. El-Sapa26 studied the axisymmetric motion of two rigid spheres in a Brinkman medium with slip surfaces. Sherief et al.28 discussed the problems of low Reynolds number micropolar fluid motion. The boundary collocation method is widely employed to resolve flow problems32.In actual instances of rotation of particles, solid walls surround the fluid in its immediate vicinityThe purpose of this study is to provide semi-analytical solutions for the axisymmetric rotation of a solid sphere within a non-concentric hollow filled by a couple stress fluids under the impact of slippage surfaces. As a result, the boundary collocation procedure is employed in the system of equations. Additionally, given various values of the relevant parameters such as the slippage, the separation, the size ratio, the couple stress, and the angular velocity ratios, the normalized torque acting on the interior solid sphere can be calculated. In general, the results showed good convergence across the parameters considered, and all the results are displayed graphically and calculated tabularly. Several studies have investigated this problem in viscous fluids and microfluidics, but this work concentrates on two concentric spheres with slippages in couple stress fluids.2:By the notion of a low Reynolds number, without the presence of body forces and body couples, the field constraints controlling the steady motion of an incompressible couple stress liquid are dictated by2Here, the constant entclass1pt{minimawhere 1 by using the second relation of ,,4), as w12 and micropolar fluids27 issues.Slippage restriction21 has suggested the boundary conditioning Eq.  to  to 20) tThe boundary conditions  can be wFrom Eq. 27\documeEmploying the obtained Eqs. , 21)–(2–(221)–2, and 2727 into , we get:28As a result of the axisymmetric particle being impacted by the fluid flow, a torque is generated that has a magnitude ofFor this simulation, it is assumed the annulus between the solid sphere, mentclass2pt{minimmentclass2pt{minimHence, the boundary conditions are linear so the principle of superposition can be applied. Thus, the field functions are represented as:The next boundary conditions are recommended for the two spherical surfaces:From  and use Accordingly, by applying the boundary conditions  and 35)35) to EqThese constants aic Eqs. –43) pro pro\docu in Eqs. –(43). ThA spherical particle of radius, All results produced using this collocation approach in the present section converge to at least five decimal places. We want to quantitatively represent the normalized torque operating on the inner solid sphere for a variety of parameter values found in the equations governing the phenomenon, the slippage entclass1pt{minimaentclass1pt{minima6. The curves show a clear relationship between the torque and the slip. In other words, the amount of torque produced increases with slippage and vice versa. Additionally, Fig. It is found from Tables entclass1pt{minimaentclass1pt{minimaentclass1pt{minimaentclass1pt{minimaFurthermore, Fig. Finally, the distribution of torque versus velocity slippage on the solid sphere for fixed values of the pertinent parameters is presented in Fig. 33 and34. Additionally, the impact of oscillation, the fractional approach and electro-osmotic can be employed in this study such as37.In this research, we study the interfacial slippage effect and the steady incompressible rotation of a couple stress fluids around a rotating sphere. Therefore, the graphs are used to give a numerical analysis of the torque operating on the inner solid sphere's surface. As a result, raising the couple stress coefficient results in an expected increase in torque. Additionally, it is established that the torque is significantly influenced by the second viscosity parameter. It elevates the torque's worth. In addition, it is shown that the size ratio and separation parameter have more significant on the couple stress fluid flow, especially for small values. Finally, it is determined that the slip parameter has a significant influence in raising the torque value. The motivation for studying flow slip boundary conditions comes from the possible applications in a variety of engineering and applied scientific fields, as well as from a serious grasp of hydrodynamics, which serves as the theoretical basis for the design and construction of nanofluidic devices. Additionally, the development of shale reservoirs depends heavily on a knowledge of slip flow behavior in the nano-porous medium. The future study can be applied to this work in the effect of permeability of porous medium and magnetic field such as in


Primary membranous nephropathy (MN) is a rare autoimmune cause of kidney failure. Observational studies have suggested some relationship between virus infection and primary MN, but the association remains unclear. The current study performed a two‑sample Mendelian randomization (MR) analysis to explore the causal association between varicella‐zoster virus (VZV) infection (chickenpox and shingles) and primary MN using genome‑wide association studies (GWASs) summary statistics. The exposure datasets containing chickenpox and shingles were obtained from the GWASs conducted by the 23andMe cohort. And summary‐level statistics for primary MN were used as the outcome dataset, comprising 2150 cases and 5829 controls from European Ancestry. The inverse variance weighted method was adopted as the main analysis. As a result, we found that both genetically determined chickenpox  and shingles  were causally associated with an increased risk of developing primary MN. In conclusion, our MR findings provided novel genetic evidence supporting the causal effect of VZV infection on primary MN. Further studies are needed to elucidate the underlying mechanisms mediating the causal association. It is caused by circulating antibodies binding to autoantigens expressed in the podocytes (such as M-type phospholipase A2 receptor thrombospondin type 1 domain-containing 7A), and then the immune complex deposit on the outer aspect of the basement membrane, finally leading to the thickening of the glomerular capillary walls and the leakage of proteins into the urine (proteinuria)4. The etiology of primary MN remains largely unknown, while both genetic and environmental factors have been found to play important roles in the pathogenesis of primary MN5. Among the environmental factors, few cases have described the occurrence of nephropathy triggered by Varicella zoster virus (VZV) infection6.Primary membranous nephropathy (MN) is a rare autoimmune cause of kidney failure, with an incidence estimated at about 12/million per year7. Primary VZV infection causes chickenpox, and then VZV becomes latent in ganglionic neurons8. As cellular immunity to VZV shrinks with age or in immuno-suppressed individuals, VZV would become reactivated, which causes herpes zoster (shingles) in tissues innervated by the involved neurons, and ultimately leads to persistent radicular pain 8. VZV infection has been found to be a trigger for several autoimmune diseases, such as systemic lupus erythematosus (SLE)9 and multiple sclerosis (MS)10, while little is known about the relationship between VZV infection and primary MN.VZV infection is common worldwide. The global new VZV infection cases in 2019 were over 83 million, and the global age‐standardized incidence of VZV infection has continued to increase in the past 3 decades11. For MR, the key step of MR is selecting eligible IVs (G), which are required to meet 3 assumptions: assumption 1 is that IVs (G) should be strongly related to the exposure (X); assumption 2 is that IVs(G) should not have direct associations with any other confounding factors (U) that may affect the outcome, and assumption 3 is that IVs (G) should not be directly related to the outcome (Y)12. Only if these assumptions are fulfilled, a causal effect of X on Y can be estimated using MR, taking G into account. There are distinct advantages of MR over observation studies. Firstly, assumption 2 and the test of horizontal pleiotropy in MR can avoid confounding factors11. Moreover, the direction of G → X → Y ensures the direction of the causal effect and can avoid reverse causation11. MR has been applied to find the causal effect of VZV infection on MS13 and Alzheimer’s disease (AD)14.Mendelian Randomization (MR) is an analysis method for non-experimental studies in genetic epidemiology, which utilizes genetic variants (G) strongly associated with the exposure (X) as instrumental variables (IVs) to assess the causal effect of exposures (X) on outcomes (Y) Fig. A11. For 15 were used as IVs to estimate the causal effect of VZV infection on primary MN16.In the current study, we aimed to apply the two‐sample MR approach to investigate the causal link between VZV infection (exposure) and primary MN (outcome). Briefly, single nucleotide polymorphisms (SNPs) strongly associated with chickenpox and shinglesThe flowchart of the study was described in Fig. 17, so we chose the largest GWASs to conduct the MR analyses. Chickenpox and shingles are both caused by the infection of VZV, therefore, we chose both chickenpox and shingles as exposure phenotypes to represent VZV infection. And primary MN was used as the outcome phenotype. Summary statistics of chickenpox and shingles were downloaded from the chickenpox and shingles GWASs conducted by the 23andMe cohort, which included 107,769 cases and 15,982 controls for chickenpox, and 16,711 cases and 118,152 controls for shingles, with more than 97% of the participants from European ancestry15. Summary‐level statistics for primary MN were obtained from the GWAS meta-analysis, comprising 2150 primary MN cases and 5829 controls16. All primary MN cases used in this study were defined by a kidney biopsy diagnosis of idiopathic MN16. Any suspected secondary cases due to drugs, malignancy, infection, or autoimmune diseases were excluded16. All datasets mentioned above were obtained from publicly available repositories. Sample collection, genotyping and quality control were conducted by the researchers who published the original publications, and the detailed methods can be found as referenced16. Detailed information on the records was listed in Supplementary Table GWASs require much larger sample sizes to achieve adequate statistical power, which guarantees the success of detecting loci strongly associated with human complex diseases12. To meet assumption 1 (IVs should be strongly related to the exposure) and assumption 3 (IVs should not be directly related to the outcome), SNPs that were significantly associated with the exposure but not with the outcome on a genome-wide basis (p < 5.0e−08) and had a Cragg–Donald F-statistic > 10 were selected as strong IVs18. However, assumption 2 (IVs should not have direct associations with any other confounding factors) was difficult in the IV selection stage because confounding factors were hard to define, therefore, it was calculated as horizontal pleiotropy in the post-MR analysis12.Eligible IVs for MR should meet 3 assumptions2 = 0.05 within the window of 10 mega-base pairs to avoid double counting and biased causal effect estimates. Next, the IVs were extracted from the outcome trait and were harmonized in both exposure and outcome GWAS datasets. In this step, palindromic SNPs with intermediate allele frequency were removed. Moreover, if a particular requested SNP was not present in the outcome GWAS dataset, then an SNP (proxy) that is in LD with the requested SNP (target) will be searched, which was defined using 1000 genomes European sample data (r2 ≥ 0.8). After these steps, the MR effect can be estimated. The inverse variance weighted (IVW) method was performed as the main analysis, which is the most efficient analysis method with valid IVs because it accounts for heterogeneity in the variant-specific causal estimates19. Moreover, additional sensitivity analyses including the simple mode, weighted mode, weighted median, and MR-Egger regression methods, were further conducted to assess the robustness of the findings19. And the leave-one-out analysis was conducted within the IVW method to assess the influence of individual variants on the observed association. Furthermore, post-MR analyses including Cochran’s Q test for heterogeneity and MR Egger Intercept test for horizontal pleiotropy were also performed20. Lastly, we computed the proportion of variance in the phenotype explained by IVs and calculated the statistical power for the MR study with a two-sided type-I error rate α = 0.0521. The main statistical analyses were conducted using TwoSampleMR (v.0.5.6) in the R package (V.4.2.2)22. The study was performed according to the STROBE MR guideline23.To ensure enough number of IVs, independent IVs were set at a threshold of linkage disequilibrium (LD) at rQ test indicated no heterogeneity across the IVs (p = 0.726)  Table . While sQ test detected some heterogeneity among the IVs  was different from the trend in the main IVW analysis (OR > 1.00 and significant) Fig. A,C, whic4. Viruses are known to trigger autoimmunity via mechanisms such as molecular mimicry, where a foreign antigen shares sequence or structural similarities with self-antigens26. Therefore, it can be speculated that after VZV infection, the immune system might mistake the VZVs as antigens, and the antigen–antibody complex will deposit on the basement membrane and ultimately lead to the development13, while further functional studies are needed. On the other hand, the innate immune response to VZV might trigger inflammatory cytokine secretion and/or cell death, which further involves the pathogenesis of primary MN27. However, further studies were warranted to clarify how VZV infection is involved in the development of primary MN.Primary MN is an autoimmune disease. The pathophysiological mechanism of primary MN is that the circulating antibodies bind to autoantigens expressed in the podocytes, leading to the thickening of the glomerular capillary walls and the leakage of proteins into the urine (proteinuria)28. VZV infection is very common in the general population, and previous studies have shown that the incidence ranges from 13 to 16 cases per 1000 persons per year and the global age‐standardized incidence of VZV infection has continued to increase in the past 3 decades8. Moreover, VZV infection is highly communicable, and spreads by the air-borne route, with an extraordinarily high transmission rate8. Fortunately, antiviral drugs and vaccines against both VZV are available and are effective in treating and preventing VZV-induced disease30. Therefore, our MR results indicated that prevention of VZV infection, such as administering the vaccine, might decrease the risk of developing primary MN. However, considering that VZV is common while primary MN is pretty rare, other factors such as immune response differences between individuals should also be noted, which needs further investigation. VZV, also known as human herpesvirus 3, is a ubiquitous alphaherpesvirus with a double-stranded DNA genomeThere were several advantages of the current study. Firstly, we used SNPs strongly associated with the exposure (chickenpox and shingles) as valid IVs to calculate the causal association from exposure to outcome (primary MN), therefore, it ensures the direction of the causal effect and can avoid reverse causation. Secondly, the test of horizontal pleiotropy can ensure the causal association between the exposure and the outcome without the effects of confounding factors. Thirdly, we applied both chickenpox and shingles as phenotypes to represent VZV infection, which confirmed the robustness of our findings.However, some limitations should be acknowledged in the current study. First, there were only 2 IVs used in analyzing the causal effect of chickenpox on primary MN, the IV-based heritability could only explain a small proportion of variance, and the statistical power of assessing the causality of chickenpox on primary MN was slightly low (0.72); therefore, GWASs with a larger sample size would be helpful. Secondly, chickenpox and shingles cases were determined by self-reported history, without confirmation of laboratory test results, which might bias the GWAS results. Therefore, GWAS conducted among laboratory-confirmed VZV infection cases was needed. Thirdly, the datasets used in the current study were all derived from the European population, more studies from other ethnic groups are needed to confirm the results. Last but not least, the underlying mechanisms mediating the causal association between VZV infection and primary MN were not investigated, and further functional studies were needed.In conclusion, our MR findings using GWAS summary statistics provided novel genetic evidence indicating the causal effect of VZV infection on primary MN. Therefore, prevention of VZV infection, such as administering the vaccine, might decrease the risk of developing primary MN. Further studies are needed to elucidate the underlying mechanisms mediating the causal association.Supplementary Figure 1.Supplementary Table 1.


The present study was the outcome of using chemicals and consumables purchased from this grant without compromising its main objectives and milestones.Funding for this work was provided by Ministry of Education, Malaysia, with Fundamental Research Grant Scheme (FRGS) with external reference number FRGS/1/2022/STG01/ UMP/02/1 and title: Investigations of Protein from the Lesser Known Tongkat Ali Plants of is changed to:Stema tuberosa and Polyalthia bullata for Their Potentials in Improving Men’s Health. The present study was the outcome of using chemicals and consumables purchased from this grant without compromising its main objectives and milestones.The authors would like to thank Ministry of Higher Education for providing financial support under Fundamental Research Grant Scheme (FRGS) No: FRGS/1/2022/STG01/UMP/02/1 (University reference RDU220110) and Universiti Malaysia Pahang for laboratory facilities. The title of the FRGS grant: Investigations of Protein from the Lesser Known Tongkat Ali Plants of


Calprotectin (CP) is a calcium and zinc binding protein that is widely measured on faecal samples but its determination in other biological fluids might be of interest. The aim of this work was to validate the measurement of CP in pleural fluid by chemiluminescence.®XL, a fully automated chemiluminescence analyzer, was used for CP quantification on pleural fluid. A validation protocol was designed using both quality control materials provided by the manufacturer and pools of pleural fluid samples. Stability, imprecision, bias, linearity, detection capability and carry over effect were evaluated.LIAISONCP was stable on pleural fluid at least one week, under refrigerated conditions, and four weeks at −80 °C. The observed intra- and inter-day imprecision was between 2.2 and 6.49 %, with a negative bias under 5.51 %. The linearity of the method was verified up to 2,000 ng/mL. The LoQ for the assay was 48.52 ng/mL. A statistically significant carry-over effect was observed after measuring CP concentrations above the upper limit of linearity, but given the observed magnitude, a clinically relevant impact should not be expected.® calprotectin assay allows reliable measurement of CP in pleural fluid.DiaSorin Liaison Calprotectin (CP) (S100A8/S100A9) is a calcium and zinc binding protein of 36 kDa, member of the S100 family of proteins .In clinical practice, fecal CP measurement  is widelHowever, fully automated, fast and reliable methods are nowadays available. The aim of this work was to validate the measurement of CP in pleural fluid by chemiluminescence, hypothesizing that this method could provide better clinical performance than ELISA.®XL , a fully automated chemiluminescence analyzer, was used for CP quantification on pleural fluid. This analyzer was designed for the quantification of fecal calprotectin so results are expressed in µg/g. Based on previous publication . The experiment was repeated with lower values of (a) close to the upper limit of the linear range, obtaining  . A statiCurrently, in clinical practice, CP levels are routinely measured just in fecal samples in the diagnosis and follow-up of patients suffering from chronic inflammatory gastrointestinal disorders .However, CP has been postulated in recent years as an emerging biomarker to monitor the follow-up of active disease in other autoimmune disorders, since CP plays a pivotal role in innate immunity . In fact®), has been described for the first time.CP also can be found in other biological fluids as effusions: it seems that in synovial fluid CP measure has interest in RA patients , 17 and In the present work, we have demonstrated that the stability of CP in pleural fluid (7 days) is even higher than in feces (72 h), and in EDTA plasma samples (4 days)  under reThe observed imprecision in pleural fluid sample pools is lower than that described by ELISA method  [The claimed linearity has been verified in LP.®) or lateral flow immunocromatographic method (Quantum Blue®)  [A slightly higher LoQ was observed in pleural fluid than in feces, that should be taken into consideration for future applications where low CP concentrations have clinical interest. Even so, LoQ by this method is lower than that described by ELISA (ELISA fCALctively) .Finally, a slight carryover effect can be observed not only when measuring very high CP concentrations, but also near the upper limit of the linearity range. However, being within the tolerance margins of analytical performance, clinically relevant differences due to carryover should not be expected.In conclusion, this study demonstrates that automated chemiluminescence assay provides better clinical performance than other methods described in previous studies.® calprotectin assay allows reliable measurement of CP in pleural fluid and the analytical performance described here encourages further evaluation of its potential usefulness in clinical practice.DiaSorin Liaison