Bile Samples Research Articles

Effect and mechanism of endoclip papilloplasty in reducing the incidence of cholelithiasis.

Endoscopic sphincterotomy (EST) is widely used to treat common bile duct stones (CBDS); however, long-term studies have revealed the increasing incidence of recurrent CBDS after EST. Loss of sphincter of Oddi function after EST was the main cause of recurrent CBDS. Reparation of the sphincter of Oddi is therefore crucial. This study aims to investigate the effectiveness and safety of endoclip papilloplasty (ECPP) for repairing the sphincter of Oddi and elucidate its mechanism. Eight healthy Bama minipigs were randomly divided into the EST group and the ECPP group at a 1:1 ratio, and bile samples were collected before endoscopy and 6months later. All minipigs underwent transabdominal biliary ultrasonography for the diagnosis of cholelithiasis 6months after endoscopy. The biliary microbiota composition and alpha and beta diversity were analyzed by 16S rRNA gene sequencing. Differential metabolites were analyzed by bile acid metabolomics to explore the predictive indicators of cholelithiasis. Three minipigs were diagnosed with cholelithiasis in the EST group, while none in the ECPP group showed cholelithiasis. The biliary Firmicutes/Bacteroidota (F/B) ratio was increased after EST and decreased after ECPP. The Chao1 and observed species index significantly decreased 6months after EST (P=0.017 and 0.018, respectively); however, the biliary α-diversity was similar before and 6months after ECPP. The β-diversity significantly differed in the EST group before and 6months after EST, as well as in the ECPP group before and 6months after ECPP (analysis of similarities [ANOSIM]: R=0.917, P=0.040; R=0.740, P=0.035; respectively). Glycolithocholic acid (GLCA) and taurolithocholic acid (TLCA) accumulated in bile 6months after EST. ECPP has less impact on the biliary microenvironment than EST and prevents duodenobiliary reflux by repairing the sphincter of Oddi. The bile levels of GLCA and TLCA may be used to predict the risk of cholelithiasis.

Analysis on characteristics and multilocus sequence typing of Clostridium perfringens in western China.

The objective of this study was to identify and analyse the distribution characteristics, toxin genotyping and antimicrobial susceptibility of Clostridium perfringens and to investigate its resistance mechanisms and genetic characteristics. The MICs of various antibiotics against C. perfringens were determined using the agar dilution method, and resistance genes and toxin genotypes were detected by PCR. Genetic relationships were analysed using MLST. WGS was conducted on the DNB system and PacBio platforms. Analysis of 36 strains of C. perfringens revealed that the major toxin types were types C and F, with 86.1% of the strains isolated from bile samples. Of these, 30.6% of the strains exhibited MDR, with resistance rates of 75.0%, 52.8% and 52.8% for penicillin, clindamycin and ampicillin, respectively; however, no resistance to metronidazole and carbapenems was observed. MLST analysis identified 29 STs, including 14 novel types. ST221 and ST498 were the dominant types. The WGS revealed that the most prevalent virulence factors are plc (100.0%), nagH (100.0%), colA (100.0%), nanJ (100.0%), entB (100%), nanH (97.0%), entA (97.0%) and nanI (90.9%). Among these factors, the primary determinants of tetracycline resistance are tetA (66.7%) and tetB (78.8%), which represent the most frequently detected antibiotic resistance genes. This study indicates that the infection rate of C. perfringens is relatively high, with the majority of isolated strains exhibiting MDR. The observed high levels of antibiotic resistance, combined with the significant genetic diversity of these strains, suggest a potential public health risk.

Detection of biliary trematodes in sheep and goats from northern and central Portugal

Fasciolosis and dicrocoeliosis are severe parasitic infections that result in substantial economic damages to livestock farming globally as a consequence of reduced productivity and viscera condemnation. Molecular tests such as polymerase chain reaction (PCR) can detect Fasciola hepatica and Dicrocoelium dendriticum DNA with high sensitivity and specificity. In this study, we aimed to assess the presence of F. hepatica and D. dendriticum by PCR-based techniques in 400 small ruminant bile samples retrieved from central Portugal. Additionally, we conducted genetic characterisation of F. hepatica and D. dendriticum in these samples. Only one of the 400 bile samples (0.25%; 95% confidence interval [CI]: 0.01–1.39) examined by PCR tested positive for F. hepatica, and two samples (0.50%, 95% CI: 0.06–1.79) tested positive for D. dendriticum. Our findings indicate a low prevalence of F. hepatica and D. dendriticum in Portuguese small ruminants, underscoring the need to investigatefactors leading to meat rejection in slaughterhouses. Implementing effective parasite control measures is crucial to minimise economic losses and improve food safety. Addressing these infections and deploying targeted control strategies can enhance livestock production sustainability and ensure safe, high-quality meat products for consumers.

Bile molecular landscape provides pathological insight and classifies signatures predictive of carcinoma of gall bladder.

Carcinoma of gall bladder (CAGB) has poor prognosis. Molecular analysis of bile could classify indicators of CAGB. Bile samples (n=87; training cohort) were screened for proteomics and metabolomics signatures of cancer detection. In bile, CAGB showed distinct proteomic (217up-, 258 downregulated) and metabolomic phenotype (111 up-, 505 downregulated, p<0.05, FC>1.5, FDR<0.01) linked to significantly increased inflammation (coagulation, arachidonic acid, bile acid) and alternate energy pathways (pentose-phosphate pathway, amino-acid, lipid metabolism) and decreased glycolysis, cholesterol metabolism, PPAR, RAS and RAP1 signaling, oxidative phosphorylation, and others compared to GS or HC (p<0.05). Bile proteins/metabolites signatures showed significant correlation (r2>0.5, p<0.05) with clinical parameters. Metabolite/protein signature-based probability of detection for CAGB (cancer) was >90% (p<0.05) with AUC>0.94. Validation of top four metabolites: Toluene, 5,6-DHET, Creatine, and Phenylacetaldehyde in separate cohorts (n=80; bile [T1] and paired plasma [T2]) showed accuracy (99%) and sensitivity/specificity (>98%) for CAGB detection. Tissue validation showed bile 5,6 DHET positively correlated with tissue PCNA (proliferation), and Caspase-3 linked to cancer development (r2>0.5, p<0.05). In conclusion, the bile molecular landscape provides critical molecular understanding and outlines metabolomic indicator panels for early CAGB detection.

A worrisome prevalence of extended-spectrum β-lactamase producers in patients with biliary obstruction and cholangitis: Phenotypic and molecular characterization of biliary Escherichia coli and Klebsiella pneumoniae isolates

Background & aimsThe alarming rise of antibiotic resistance presents a substantial and worrisome issue within the context of biliary obstruction, specifically in the treatment of cholangitis. This abovementioned scenario underscores the critical importance of addressing extended spectrum β-lactamase (ESBL) producers in the biliary system to adequately tackle cholangitis using third-generation cephalosporins. Hence, we aimed to determine the frequency of ESBL and carbapenemases among biliary Escherichia coli and Klebsiella pneumoniae isolated from patients with biliary obstruction. MethodsIn this cross-sectional study, bile samples were collected via aspiration from patients diagnosed with biliary obstruction during the endoscopic retrograde cholangiopancreatography (ERCP) procedure. Subsequent culturing of these samples was performed, followed by phenotypic and molecular assessments for the detection of ESBL- and carbapenemase-producing strains of E. coli and K. pneumoniae. ResultsApproximately 23.5 % of patients with biliary obstruction harbored biliary ESBL-producers, with the majority (70.2 %) being diagnosed with cholangitis. Moreover, 2.1 % of patients had biliary carbapenemase-producing K. pneumoniae strains. Molecular analysis confirmed the high prevalence of blaCTX-M and blaTEM in E. coli, and blaTEM and blaSHV in K. pneumoniae. Additionally, the presence of biliary K. pneumoniae harboring blaKPC, blaNDM, and blaIMP was observed. ConclusionOur study reveals a noteworthy observation that over half of patients experiencing biliary obstruction harbor ESBL-producing bacteria in their biliary tract. Notably, we discovered a significant link between ESBL producers and the risk of cholangitis. These findings raise important concerns regarding the suitability of employing third-generation cephalosporins as initial treatment for cholangitis and other similar biliary infections.

Енергетичний обмін у печінці щурів із паркінсонічним синдромом, індукованим ротеноном, при дії метаніндіазенону

Parkinson’s disease (PD) is symptomatically characterized by motor disorders in the human body, which are associated with the degeneration of dopamine neurons in the substantia nigra of the midbrain. It is widely recognized that mitochondrial dysfunction occurs in dopaminergic neurons, leading to their death and the development of this pathology. Recently, benzodiazepine derivatives have been found to have a neuroprotective effect against damage of dopaminergic neurons in an experimental model of Рarkinsonian syndrome (PS). The aim of this study was to investigate the effect of a novel molecule, named methanіdiazenone, on energy metabolism in the liver of rats induced by rotenone. To achieve the set goal, the concentration of ATP, ADP, AMP, hypoxanthine аnd xanthine were determined in the bile samples of rats with PS by the method of thin-layer chromatography. The obtained data indicate that under the influence of rotenone compared to the control values the content of ATP in bile decreased by 40%, while the levels of AMP, xanthine and hypoxanthine, on the contrary, increased by 87,5%, 55,6%, and 25%, respectively. Our findings suggested that the ratio of AMP/ATP, which is an important indicator of the functional state of mitochondria, under the influence of rotenone increased by 200% compared to the control. Metanindiazenone in a dose of 1,0 and 2,0 mg/kg normalized all studied parameters of purine metabolism. The presented results show that methanindiazenone significantly improves purine metabolism in the liver of rats with PS induced by rotenone, indicating the normalization of mitochondrial function in hepatocytes. Thus, methanindiazenone may be recommended for clinical trials regarding its use in combination with other drugs to treat Parkinson’s disease and possibly other neurodegenerative diseases, in which mitochondrial dysfunction occurs.

Comprehensive Analysis of Bile Medicines Based on UHPLC-QTOF-MSE and Machine Learning.

Based on UHPLC-QTOF-MSE analysis and quantized processing, combined with machine learning algorithms, data modeling was carried out to realize digital identification of bear bile powder (BBP), chicken bile powder (CIBP), duck bile powder (DBP), cow bile powder (CBP), sheep bile powder (SBP), pig bile powder (PBP), snake bile powder (SNBP), rabbit bile powder (RBP), and goose bile powder (GBP). First, 173 batches of bile samples were analyzed by UHPLC-QTOF-MSE to obtain the retention time-exact mass (RTEM) data pair to identify bile acid-like chemical components. Then, the data were modeled by combining support vector machine (SVM), random forest (RF), artificial neural network (ANN), gradient boosting (GB), AdaBoost (AB), and Naive Bayes (NB), and the models were evaluated by the parameters of accuracy (Acc), precision (P), and area under the curve (AUC). Finally, the bile medicines were digitally identified based on the optimal model. The results showed that the RF model constructed based on the identified 12 bile acid-like chemical constituents and random forest algorithm is optimal with ACC, P, and AUC > 0.950. In addition, the accuracy of external identification verification of 42 batches of bile medicines detected at different times is 100.0%. So based on UHPLC-QTOF-MSE analysis and combined with the RF algorithm, it can efficiently and accurately realize the digital identification of bile medicines, which can provide reference and assistance for the quality control of bile medicines. In addition, hyodeoxycholic acid, glycohyodeoxycholic acid, and taurochenodeoxycholic acid, and so forth are the most important bile acid constituents for the identification of nine bile medicines.

Concentrations of ceftazidime and avibactam in bile fluid-a prospective phase IIb study.

The rise in carbapenem-resistant bacteria and the limited number of effective antibiotics pose a major health-care threat. The combination of ceftazidime (CAZ) and avibactam (AVI) represents an approved treatment option for carbapenem-resistant intra-abdominal infections. However, data on the pharmacokinetic profile of AVI in the hepatobiliary compartment is lacking. To provide clinical in vivo data on the concentration of AVI in bile fluid as a surrogate for hepatobiliary excretion. A single dose of 2000/500 mg CAZ/AVI was administered prolonged over 2 h to 10 patients prior to abdominal surgery, with bile samples available in nine patients in this phase IIb study (DRKS-ID: DRKS00023533). Antibiotic concentrations in plasma (0-8 h), bile (after resection) and pharmacodynamic parameters were determined. The mean concentration across individuals in bile was 33.5 mg/L (±20.5 mg/L) for CAZ and 7.1 mg/L (±3.5 mg/L) for AVI, resulting in bile/plasma ratios of 0.58 (±0.26) and 0.61 (±0.18). The Cmax in plasma was 87.2 mg/L (±25.0 mg/L) for CAZ and 18.6 mg/L (±6.29 mg/L) for AVI, with AUC0-∞ values of 351 h·mg/L (±104 h·mg/L) and 72.1 h·mg/L (±32.1 h·mg/L), respectively. Plasma concentrations of both CAZ and AVI remained more than 50% of the dosing interval above the minimum inhibitory concentrations (T>MIC > 50%; MICCAZ = 8 mg/L, MICAVI = 1 mg/L) in all patients. No antibiotic-associated side effects were reported during the 30-day follow-up. The concentrations of CAZ and AVI in bile suggest their potential as a valuable therapeutic option for multi-resistant biliary infections.

Optimal preservation condition for the extraction of cell-free DNA from bile of patients with biliary tract cancer.

Patients with biliary tract cancer (BTC) often have dismal outcomes due to the poor performance of traditional methods for early diagnosis. Recently, bile cell-free DNA (cfDNA) has been reported as a potential liquid biopsy material for BTC diagnosis. However, bile is a complex alkaline aqueous medium, and the proper storage conditions for bile remain to be explored. The aim of this study is to explore the effects of storing bile under various conditions on the stability of bile cfDNA and to determine the optimal conditions, thereby establishing a foundation for the subsequent application of bile cfDNA in liquid biopsy for early diagnostic and prognosis monitoring of patients with malignant BTC. We evaluated the storage temperature and storage time for the preservation of bile samples. Bile samples were collected in cfDNA tubes with protectant covered inside or regular tubes without, and the stability of bile cfDNA was analyzed during 10 days at room temperature (RT) or after 2 months of storage at low temperatures. Bile cfDNA remained stable for bile samples being collected with cfDNA tubes and stored for 10 days at RT, while degraded with time for the case with regular tubes. When bile samples were collected with cfDNA tubes and stored for 2 months at 4 ℃, bile cfDNA remained stable, however, if collected with regular tubes, bile cfDNA exhibited a slight loss of integrity. No significant difference was observed for 2 months storage at -20 or -80 ℃. Our findings suggested that for bile cfDNA research, bile samples should be collected with cfDNA tubes and it can be transported for short-term shipment at RT, and could be stored at 4 ℃ with cfDNA tubes, or frozen at -20 ℃ with regular tubes.

Detection of H1N1 Influenza Virus in the Bile of a Severe Influenza Mouse Model.

Influenza virus infection may lead to fatal complications including multi-organ failure and sepsis. The influenza virus was detected in various extra-pulmonary organs in autopsy studies during the 2009 pandemic. However, limited research has been conducted on the presence of viral particle or viral components in the peripheral blood. We established a mouse model for severe H1N1 influenza. The bile and blood samples were collected over time and inoculated into embryonated chicken eggs. We detected live influenza virus in bile and blood samples in early infection. Immunofluorescence showed influenza viral components in the liver tissue. No live virus was isolated in the bile in mice intragastrically administered with influenza virus, indicating that the virus was spread from the blood stream. Targeted metabolomics analysis of bile acid and liver tissues showed that a secondary bile acid (3-dehydrocholic acid) was decreased after influenza H1N1 infection. Genes related with fatty acid metabolism and bile secretion pathways were down-regulated in liver after influenza virus infection. Our study indicated that influenza virus viremia is present in severe influenza, and that the liver is a target organ for influenza viral sepsis.

Integration of bile proteomics and metabolomics analyses reveals novel insights into different types of gallstones in a high-altitude area

BackgroundTo explore the pathogenesis of different subtypes of gallstones in high-altitude populations from a molecular perspective.MethodsWe collected bile samples from 20 cholesterol gallstone disease (CGD) patients and 20 pigment gallstone disease (PGD) patients. Proteomics analysis was performed by LC/MS DIA, while metabolomics analysis was performed by UPLC- Q-TOF/MS.ResultsWe identified 154 up-regulated and 196 down-regulated differentially expressed proteins, which were significantly enriched in neurodegenerative diseases, energy metabolism, amino acid metabolism etc. In metabolomics analysis, 20 up-regulated and 63 down-regulated differentially expressed metabolites were identified, and they were significantly enriched in vitamin B6 metabolism. Three pathways of integrated proteomics and metabolomics were significantly enriched: porphyrin and chlorophyll metabolism, riboflavin metabolism and aminoacyl-tRNA biosynthesis. Remarkably, 7 differentially expressed proteins and metabolites showed excellent predictive performance and were selected as potential biomarkers.ConclusionThe findings of our metabolomics and proteomics analyses help to elucidate the underlying mechanisms of gallstone formation in high-altitude populations.

Serological and Molecular Survey of Hepatitis E Virus in Small Ruminants from Central Portugal

Hepatitis E virus (HEV) is currently recognized as an emerging problem and a growing concern for public health in developed countries, with HEV infections mainly attributable to foodborne transmission of HEV-3. The zoonotic HEV genotype 3 infects a wide range of mammalian hosts, with swine considered as the primary host. This study investigates the occurrence of HEV among small ruminants in Portugal. The primary aim of the present research was to evaluate the circulation and the potential for HEV infection among sheep and goats. A total of 400 bile samples and 493 blood samples were collected from sheep and goats at a slaughterhouse in the center region of Portugal, between January 2022 and March 2023. The HEV RNA detection in bile samples was performed using a nested broad-spectrum RT-PCR targeting the ORF1 region. Serological analysis to detect anti-HEV antibodies was conducted using a commercial double-antigen sandwich multi-species ELISA. The HEV RNA was not detected in any bile samples using the nested broad-spectrum RT-PCR. Serological analysis revealed an overall HEV antibody seroprevalence of 2% (10/493, 95% CI: 0.98–3.70) among the small ruminants, namely 2.2% in goats and 2.0% in sheep. Curiously, no statistically significant association among the factors, age, sex and species and HEV seroprevalence was observed. Although HEV RNA was not detected in the bile of sheep and goats, this study the evidence of seroprevalence in these small ruminant species. Further research could provide additional insights into the factors influencing HEV transmission dynamics in small ruminants in Portugal and its potential implications for public health.

How contaminated is flatfish living near World Wars’ munition dumping sites with energetic compounds?

Seas worldwide are threatened by an emerging source of pollution as millions of tons of warfare materials were dumped after the World Wars. As their metal shells are progressively corroding, energetic compounds (EC) leak out and distribute in the marine environment. EC are taken up by aquatic organisms and pose a threat to both the marine ecosphere and the human seafood consumer because of their toxicity and potential carcinogenicity. Here, sediment samples and fish from different locations in the German North Sea of Lower Saxony were examined to determine whether EC transfer to fish living close to munition dumping areas. EC were found in sediments with a maximum concentration of 1.5 ng/kg. All analyzed fish muscle tissues/fillets and bile samples were positive for EC detection. In bile, the max. EC concentrations ranged between 0.25 and 1.25 ng/mL. Interestingly, while detected TNT metabolites in the muscle tissues were in concentrations of max. 1 ng/g (dry weight), TNT itself was found in concentrations of up to 4 ng/g (dry weight). As we found considerable higher amounts of non-metabolized TNT in the fish muscle, rather than TNT metabolites, we conclude an additional absorption route of EC into fish other than per diet. This is the first study to detect EC in the edible parts of fish caught randomly in the North Sea.

Acute cholangitis: a state-of-the-art review.

Acute cholangitis is a potentially life-threatening bacterial infection of the intra and/or extrahepatic bile ducts. It remains the second and third cause of community-acquired and hospital-acquired bacteremia, respectively, and is associated with mortality rates of up to 15%, despite advances in broad-spectrum antimicrobial therapy and improved access to emergency biliary tract decompression procedures. Even though not much has changed in recent years in terms of diagnosis or treatment, new data have emerged regarding multidrug-resistant bacteria that serve as etiologic agents of cholangitis. Moreover, different approaches in antibiotic regimes depending on severity grading and bile sample cultures as well as novel minimally invasive endoscopic procedures that can help when consecrated treatments such as endoscopic retrograde cholangiopancreatography (ERCP) fail, cannot be performed, or are unavailable have been proposed. This state-of-the-art review aims to offer a complete and updated assessment of the epidemiology, novel diagnostic and therapeutic methods, complications, and prognostic variables of acute cholangitis. The authors will review the prognostic implications of unusual complications, the relevance of regular bile samples and antibiograms, and their new role in guiding antibiotic therapy and limiting antibiotic resistance to present an organized and comprehensive approach to the care of acute cholangitis.

COMPARATIVE SENSITIVITY OF FAECAL AND BILE SAMPLES IN THE DIAGNOSIS OF LIVER FLUKE INFECTION IN SHEEP AND GOATS IN ZARIA, NIGERIA

Liver fluke disease (also known as liver rot) is an economically important disease of ruminant's worldwide, causing significant morbidity and grave economic loss. In a study to determine the degree of sensitivity of bile sample and faecal sample to the detection of liver flukes (Fasciola gigantica and Dicrocoelium hospes) infection in sheep and goats slaughtered in Zaria. A total of 200 samples each of gall bladders and faeces were collected from 174 goats and 26 sheep. The contents of the gall bladder and faecal samples were analyzed using sedimentation technique to isolate the eggs of these parasites and possibly the adult stages. The percentage sensitivity for bile and faecal samples were found to be approximately 90% and 63.2% respectively as more of the parasites were isolated from bile than faeces. It was then concluded that compared to examination of faeces, bile analysis is a more reliable method for the diagnoses of liver fluke infection in small ruminants around the study area.

Distinctive protein expression in elderly livers in a Sprague–Dawley rat model of normothermic ex vivo liver machine perfusion

BackgroundLiver grafts are frequently declined due to high donor age or age mismatch with the recipient. To improve the outcome of marginal grafts, we aimed to characterize the performance of elderly vs. young liver grafts in a standardized rat model of normothermic ex vivo liver machine perfusion (NMP).MethodsLivers from Sprague–Dawley rats aged 3 or 12 months were procured and perfused for 6 h using a rat NMP system or collected as a reference group (n = 6/group). Tissue, bile, and perfusate samples were used for biochemical, and proteomic analyses.ResultsAll livers cleared lactate during perfusion and continued to produce bile after 6 h of perfusion (614 mg/h). Peak urea levels in 12-month-old animals were higher than in younger animals. Arterial and portal venous pressure, bile production and pH did not differ between groups. Proteomic analysis identified a total of 1477 proteins with oxidoreductase and catalytic activity dominating the gene ontology analysis. Proteins such as aldehyde dehydrogenase 1A1 and 2-Hydroxyacid oxidase 2 were significantly more present in livers of older age.ConclusionsYoung and elderly liver grafts exhibited similar viability during NMP, though proteomic analyses indicated that older grafts are less resilient to oxidative stress. Our study is limited by the elderly animal age, which corresponds to mature but not elderly human age typically seen in marginal human livers. Nevertheless, reducing oxidative stress could be a promising therapeutic target in the future.

Recent solid-phase microextraction-based analytical approaches for the profiling of biliary bile acids in pre-transplant assessments of liver grafts subjected to normothermic ex vivo liver perfusion

BackgroundLiver transplantation is the definitive treatment for end-stage liver failure, but the scarcity of donor organs remains a significant challenge. Leveraging organs from extended criteria donors (ECD) offers a potential avenue to address worldwide shortages, though these organs are more susceptible to post-reperfusion injury. This study explores the use of normothermic ex vivo liver perfusion (NEVLP) as a method for organ preservation – an approach that sustains liver metabolism and facilitates pre-transplant assessments of organ viability via bile analysis. The focal point of this study revolves on the development of analytical methods for determining the bile acid profile throughout the peritransplantation period as a potential indicator of liver function and viability. ResultsThe study optimized and validated a high-throughput analytical method to quantify selected bile acids in bile samples using a thin-film microextraction-liquid chromatography-mass spectrometry (TFME-LC-MS) platform. Furthermore, it introduced a solid-phase microextraction-microfluidic open interface-mass spectrometry (SPME-MOI-MS) method for rapid direct analysis of bile acid isobar groups. In the animal study, discernible variations in the concentrations of specific bile acids were observed between donors after circulatory death (DCD) and heart-beating donors (HBD), particularly following normothermic perfusion and reperfusion. Noteworthy fluctuations in individual bile acid concentrations were observed throughout the entire organ transplantation process, with taurocholic acid (TCA), glycocholic acid (GCA), and glycochenodeoxycholic acid (GCDCA) emerging as promising indicators of organ quality. The efficacy of the SPME-MOI-MS platform in corroborating these trends highlights its potential for real-time bile acid analysis during liver transplantation procedures. SignificanceOur findings underscore the efficacy of NEVLP in tandem with advanced bile acid analysis methods as a reliable strategy for pre-transplant assessments of organ viability, potentially increasing the use of ECD organs and reducing organ shortages. The ability to monitor bile acid profiles in real-time provides crucial insights into liver function and ischemic injury, making significant strides in improving transplant outcomes and patient survival rates.

Quality Assessment by Bile Composition in Normothermic Machine Perfusion of Rat Livers.

Background: The persistent challenge of organ scarcity in liver transplantation leads to an escalating dependence on organs obtained from extended criteria donors (ECD). Normothermic machine perfusion (NMP) is used for improved preservation. Due to the mimicked invivo conditions during normothermic machine perfusion, the liver is metabolically active, which allows quality assessment during perfusion. Bile seems to be of rising interest in clinical studies, as it is easily collectible for analysis. As there are currently no data on biliary bile acids during NMP, the primary objective of this study was to use our experimental rodent NMP model to assess changes in bile composition through organ damage during perfusion to inform clinical evaluation of donor organs during NMP. Methods: Thirty livers from male Sprague-Dawley rats in five groups underwent 6 h of NMP using either erythrocyte-supplemented DMEM or Steen solution, with or without 30 min of warm ischemia time (WIT). We conducted regular measurements of AST, ALT, LDH, and urea levels in the perfusate at 3-hour intervals. Bile samples were analyzed for biliary pH, LDH, and gamma glutamyltransferase, as well as biliary bile acids via mass spectrometry and UHPLC. Results: Compared with regular livers, liver injury parameters were significantly higher in our donation after circulatory death (DCD) model. Bile production was significantly reduced in livers exposed to WIT, and the bile showed a significantly more alkaline pH. This correlated with the concentration of total bile acids, which was significantly higher in livers experiencing WIT. However, regular livers produced a higher total amount of biliary bile acids during perfusion. Taurocholic acid and its metabolites were most prominent. Secondary bile acids were significantly reduced during perfusion due to the missing enterohepatic circulation. Conclusions: WIT-induced liver injury affects bile composition within our small-animal NMP model. We hypothesize this phenomenon to be attributed to the energy-driven nature of bile secretion, potentially explaining why DCD livers produce less, yet more concentrated, bile. Our results may inform clinical studies, in which biliary bile acids might have a potential as a quantifiable viability marker in human NMP liver transplantation studies.

Alterations of the bile microbiome is associated with progression-free survival in pancreatic ductal adenocarcinoma patients

BackgroundPatients with pancreatic ductal adenocarcinoma (PDAC) display an altered oral, gastrointestinal, and intra-pancreatic microbiome compared to healthy individuals. However, knowledge regarding the bile microbiome and its potential impact on progression-free survival in PDACs remains limited.MethodsPatients with PDAC (n = 45), including 20 matched pairs before and after surgery, and benign controls (n = 16) were included prospectively. The characteristics of the microbiomes of the total 81 bile were revealed by 16 S-rRNA gene sequencing. PDAC patients were divided into distinct groups based on tumor marker levels, disease staging, before and after surgery, as well as progression free survival (PFS) for further analysis. Disease diagnostic model was formulated utilizing the random forest algorithm.ResultsPDAC patients harbor a unique and diverse bile microbiome (PCoA, weighted Unifrac, p = 0.038), and the increasing microbial diversity is correlated with dysbiosis according to key microbes and microbial functions. Aliihoeflea emerged as the genus displaying the most significant alteration among two groups (p < 0.01). Significant differences were found in beta diversity of the bile microbiome between long-term PFS and short-term PFS groups (PCoA, weighted Unifrac, p = 0.005). Bacillota and Actinomycetota were identified as altered phylum between two groups associated with progression-free survival in all PDAC patients. Additionally, we identified three biomarkers as the most suitable set for the random forest model, which indicated a significantly elevated likelihood of disease occurrence in the PDAC group (p < 0.0001). The area under the receiver operating characteristic (ROC) curve reached 80.8% with a 95% confidence interval ranging from 55.0 to 100%. Due to the scarcity of bile samples, we were unable to conduct further external verification.ConclusionPDAC is characterized by an altered microbiome of bile ducts. Biliary dysbiosis is linked with progression-free survival in all PDACs. This study revealed the alteration of the bile microbiome in PDACs and successfully developed a diagnostic model for PDAC.

A comprehensive study of Chuanwang Xiaoyan capsule based on characterization of chemical constituents in vitro and in vivo using ultra-high-performance liquid chromatography-quadrupole-Exactive Orbitrap mass spectrometry and pharmacokinetic study of multiple components in rats using ultra-high-performance liquid chromatography-triple quadrupole-tandem mass spectrometry.

Chuanwang Xiaoyan (CWXY) capsule is primarily used to treat a variety of acute and chronic inflammations, including acute and chronic pharyngitis and tonsillitis. However, a systematic study of its chemical constituents is still not available. This study evaluated the chemical constituents in vitro and metabolite profiles in vivo of CWXY using ultra-high-performance liquid chromatography (UHPLC) coupled with Q-Exactive Orbitrap mass spectrometry, and the pharmacokinetic behaviors of the nine main components in rats were detected using ultra-high-performance liquid chromatography-triple quadrupole-mass spectrometry (UPLC-QQQ-MS/MS). A total of 92 chemical constituents in CWXY were preliminarily identified in vitro. After oral administration to rats, 56 prototype components and 128 metabolites of CWXY were detected in the biological samples of rat plasma, urine, bile, and feces. Of these prototype components and metabolites, seven new compounds, namely M15, M16, M25, M30, M31, M71, and M128, were detected for the first time. The quantitation method of nine components in rat plasma was developed using a pharmacokinetic study. To the best of our knowledge, this study was the first to investigate the pharmacokinetic behavior of triumbelletin.