INFOGEST static in vitro simulation of gastrointestinal food digestion.

Developing a mechanistic understanding of the impact of food structure and composition on human health has increasingly involved simulating digestion in the upper gastrointestinal tract. These simulations have used a wide range of different conditions that often have very little physiological relevance, and this impedes the meaningful comparison of results. The standardized protocol presented here is based on an international consensus developed by the COST INFOGEST network. The method is designed to be used with standard laboratory equipment and requires limited experience to encourage a wide range of researchers to adopt it. It is a static digestion method that uses constant ratios of meal to digestive fluids and a constant pH for each step of digestion. This makes the method simple to use but not suitable for simulating digestion kinetics. Using this method, food samples are subjected to sequential oral, gastric and intestinal digestion while parameters such as electrolytes, enzymes, bile, dilution, pH and time of digestion are based on available physiological data. This amended and improved digestion method (INFOGEST 2.0) avoids challenges associated with the original method, such as the inclusion of the oral phase and the use of gastric lipase. The method can be used to assess the endpoints resulting from digestion of foods by analyzing the digestion products (e.g., peptides/amino acids, fatty acids, simple sugars) and evaluating the release of micronutrients from the food matrix. The whole protocol can be completed in ~7 d, including ~5 d required for the determination of enzyme activities.

Assessing the impact of waste water effluents on native fish species from a semi-arid region, NE Spain.

Mediterranean rivers are strongly affected by pollution and water scarcity. Over the summer period, urban and industrial effluents arrive into the rivers with little dilution. In order to assess the water quality, two native fish species, Barbus meridionalis and Squalius laietanus, were collected from six sites along the Ripoll River (Spain). PAH metabolites, alkylphenols (nonylphenol and octylphenol) and the musk galaxolide levels were determined in bile. 7-Ethoxyresorufin-O-deethylase (EROD) and 7-benzyloxy-4-trifluoromethyl-coumarin O-debenzyloxylase (BFCOD) were measured as metabolic biomarkers, and the activity of CYP19 aromatase was determined in the ovaries of B. meridionalis as a biomarker of endocrine disruption. The analysis of bile indicated that fish from the lower course of the river were highly exposed to different pollutants. Accordingly, a significant induction of EROD (9 to 10-fold) and BFCOD (3 to 5-fold) activities were detected in both fish species together with an increased aromatase activity in females of B. meridionalis from the most polluted sites. Considering that sewage treatment plant (STP) effluents are essential for maintaining environmental flows in small Mediterranean rivers, this study highlights the need to improve the efficiency of STPs to protect fish health.

In vitro digestibility of highly concentrated methylcellulose O/W emulsions: rheological and structural changes.

The changes in structure during the digestion of highly concentrated methyl cellulose (MC) O/W emulsions and of hydrated MC were investigated. The effect of human saliva and in vitro stomach digestion was attributed to a dilution effect, rather than to pH or pepsin activity. After in vitro intestine incubation, a decrease in viscoelasticity and an increase in fat globule size were observed. The fat released after the digestion of the MC emulsion was 49.8% of the initial fat, indicating the existence of a big physical impediment. In comparison with an O/W whey protein emulsion with fat content equal to the fat released during the MC emulsion digestion, a 12% reduction in free fatty acid formation was found, which indicates that the decrease in fat bioaccessibility in the MC emulsion should be attributed not only to a physical effect against fat release but also to a further impediment related to the fat digestion process. Fat released quantification informs about the physical retention of fat in the emulsion matrix structure. Enzymes may not act if fat is not released and solubilized. Free fatty acid quantification is the real indicator of fat digestion, but contrary to the total fat released, it is affected by a wide variety of enzymatic factors, which should be considered for the correct comparison of systems of different properties, for example systems where the amount of fat release during the digestion may be different or initially unknown.

Changes of activity and isoforms of alkaline sphingomyelinase (nucleotide pyrophosphatase phosphodiesterase 7) in bile from patients undergoing endoscopic retrograde cholangiopancreatography.

BACKGROUND: Alkaline sphingomyelinase (NPP7) is an ecto-enzyme expressed in intestinal mucosa, which hydrolyses sphingomyelin (SM) to ceramide and inactivates platelet activating factor. It is also expressed in human liver and released in the bile. The enzyme may have anti-tumour and anti-inflammatory effects in colon and its levels are decreased in patients with colon cancer and ulcerative colitis. Active NPP7 is translated from a transcript of 1.4 kb, whereas an inactive form from a 1.2 kb mRNA was found in colon and liver cancer cell lines. While the roles of NPP7 in colon cancer have been intensively studied, less is known about the function and implications of NPP7 in the bile. The present study examines the changes of NPP7 in bile of patients with various hepatobiliary diseases. METHODS: Bile samples were obtained at endoscopic retrograde cholangiopancreatography (ERCP) in 59 patients with gallstone, other benign disease, tumour, and primary sclerosing cholangitis (PSC). The NPP7 activity was determined. The appearance of the 1.4 and 1.2 kb products in the bile was examined by Western blot. The results were correlated to the diseases and also plasma bilirubin and alkaline phosphatase. RESULTS: NPP7 activity in the tumour group was significantly lower than in the gallstone group (p < 0.05). The activity in the tumour plus PSC group was also lower than in gallstone plus other benign disease group (p < 0.05). Within the tumour group NPP7 activity was lowest in cholangiocarcinoma patients, being only 19% of that in gallstone patients. Bilirubin correlated inversely to NPP7 and was higher in the tumour than in the gallstone group. Western blot identified both the 1.4 kb and the 1.2 kb products in most bile samples. The density ratio for the 1.4/1.2 kb products correlated to NPP7 activity significantly. Two patients (one PSC and one cholangiocarcinoma) lacking NPP7 activity had only the 1.2 kb form in bile. CONCLUSION: NPP7 activity and the ratio of 1.4/1.2 kb products in bile are significantly decreased in malignancy, particularly in cholangiocarcinoma. The implications of the finding in diagnosis of cholangiocarcinoma and 1.2 kb product in hepatobiliary diseases require further investigation.

Viability and resistance of lactobacilli isolated from cocoa fermentation to simulated gastrointestinal digestive steps in soy yogurt.

To study the potential probiotic characteristics such as decrease of pH, microbial viability, and tolerance to simulated digestive steps of fermented soy beverage ("soy yogurt") produced with lactobacilli isolated from cocoa fermentation (Lactobacillus fermentum TcUESC01 and Lactobacillus plantarum TcUESC02) during fermentation and refrigerated storage. The sensory acceptance of the yogurts was also tested. Samples of soy yogurt produced with L. fermentum TcUESC01 or L. plantarum TcUESC02 were collected during fermentation (0, 4, 8, and 12 h) and refrigerated storage (1, 9, 18, and 27 d), and submitted to pH and bacterial viability determinations. Tolerance to simulated digestion steps was done with refrigerated storage samples at 9 °C. Simulated digestion was performed in 3 successive steps: exposure to pepsin-HCl solution, bile shock, and simulated small intestinal juice. During storage, a decrease in pH and lactobacillus viability was observed. L. fermentum TcUESC01 showed to be more resistant than L. plantarum TcUESC02 to simulated gastrointestinal digestion. All soy yogurts showed acceptable hedonic scores (greater than 5 in a 9-point hedonic scale ranging from "like extremely" to "dislike extremely") in sensory evaluation for flavor, aroma, color, consistency, and overall impression. L. plantarum TcUESC02 and, especially, L. fermentum TcUESC01 showed potential probiotic characteristics when considering pH, cell viability, and tolerance to simulated digestive steps and did not affect the sensory characteristics when supplemented to soy yogurt during storage.

Metabolic enzyme activities and drug excretion in the small intestine and in the liver in the rat.

The aim of these experiments was the investigation of the correlation between the metabolic enzyme activities and the intestinal and hepatic excretion of p-nitrophenol (PNP) and its metabolites (PNP-glucuronide: PNP-G and PNP-sulfate: PNP-S) in the same group of rats (n = 10). A jejunal loop was perfused with isotonic medium containing PNP in a concentration of 500 µM. The samples were obtained from the luminal perfusion medium and from the bile. For enzyme assays tissue samples were obtained from the liver and jejunum at the end of experiments. Significant differences were calculated by the Student's t-test. The activity of UDP-glucuronyltransferase and sulfotransferase was about three times higher in the liver than in the small intestine. The activity of the ß-glucuronidase was about six times higher, the activity of the arylsulfatase was approximately seven times greater in the liver than in the jejunum. No significant difference was found between the luminal appearance and the biliary excretion of PNP-G. Contrary to these findings, the biliary excretion of PNP-S was significantly higher than the luminal appearance of PNP-sulfate. It can be concluded that no direct correlation exists between the activity of metabolic enzymes and the excretion rate of PNP-metabolites in the liver and in the jejunal segment of the small intestine.

Endocrine disrupting chemicals in fish bile: a rapid method of analysis using English sole (Parophrys vetulus) from Puget Sound, WA, USA.

This study describes a recently developed and rapid method to measure bisphenol A (BPA), 17ß-estradiol (E2) and 17α-ethynylestradiol (EE2) in bile of fish using enzymatic hydrolysis of samples followed by solid-phase extraction and ultra-performance liquid chromatography with tandem mass spectrometry. The limits of quantitation (LOQ) for BPA, EE2 and E2 were 6.3ngmL(-1), 12.5ngmL(-1) and 6.3ngmL(-1), respectively. These compounds were analyzed in bile of male English sole (Parophrys vetulus) collected from urban and non-urban sites in Puget Sound, WA, USA. The BPA and E2 concentrations (and occurrence) ranged from

Cancer detection by ubiquitin carboxyl-terminal esterase L1 methylation in pancreatobiliary fluids.

AIM: To evaluate the utility of measuring epigenetic alterations in pancreatic and biliary fluids in determining molecular markers for pancreatobiliary cancers. METHODS: DNA was extracted from undiluted pancreatic and biliary fluids. As a surrogate for a genome-wide hypomethylation assay, levels of long interspersed nuclear element-1 (LINE-1) methylation were analyzed using bisulfite pyrosequencing. CpG island hypermethylation of 10 tumor-associated genes, aryl-hydrocarbon receptor repressor, adenomatous polyposis coli, calcium channel, voltage dependent, T type α1G subunit, insulin-like growth factor 2, O-6-methyl-guanine-DNA methyltransferase, neurogenin 1, CDKN2A, runt-related transcription factor 3 (RUNX3), secreted frizzled-related protein 1, and ubiquitin carboxyl-terminal esterase L1 (UCHL1), was analyzed using MethyLight. To examine the role of CpG methylation and histone deacetylation in the silencing of UCHL1, human gallbladder carcinoma cell lines and pancreatic carcinoma cell lines were treated with 2 or 5 µmol/L 5-AZA-dC for 72 h or 100 nmol/L Trichostatin A for 24 h. After the treatment, UCHL1 expression was analyzed by real-time reverse transcription-polymerase chain reaction. RESULTS: Pancreatobiliary cancers exhibited significantly lower LINE-1 methylation levels in pancreatic and biliary fluids than did noncancerous pancreatobiliary disease (58.7% ± 4.3% vs 61.7% ± 2.2%, P = 0.027; 53.8% ± 6.6% vs 57.5% ± 1.7%, P = 0.007); however, LINE-1 hypomethylation was more evident in pancreatic cancer tissues than in pancreatic fluids (45.4% ± 5.5% vs 58.7% ± 4.3%, P < 0.001). CpG island hypermethylation of tumor-associated genes was detected at various frequencies, but it was not correlated with LINE-1 hypomethylation. Hypermethylation of the UCHL1 gene was cancer-specific and most frequently detected in pancreatic (67%) or biliary (70%) fluids from patients with pancreatobiliary cancer. As a single marker, hypermethylation of the UCHL1 gene in pancreatic and biliary fluids was most useful for the detection of pancreatic and pancreatobiliary cancers, respectively (100% specificity). Hypermethylation of the UCHL1 and RUNX3 genes in pancreatic and biliary fluids was the most useful combined marker for pancreatic (87% sensitivity and 100% specificity) and pancreatobiliary (97% sensitivity and 100% specificity) cancers. Treatment with a demethylating agent, 5-AZA-2'-deoxycytidine, restored UCHL1 expression in pancreatobiliary cancer cell lines. CONCLUSION: Our results suggest that hypermethylation of UCHL1 and RUNX3 in pancreatobiliary fluid might be useful for the diagnosis of pancreatobiliary cancers.

Release of EPA and DHA from salmon oil - a comparison of in vitro digestion with human and porcine gastrointestinal enzymes.

In the present study, we hypothesised whether in vitro digestion of salmon oil would release different amounts of PUFA depending on the origin of the lipolytic enzymes used. For this purpose, in vitro digestion of salmon oil (SO) was performed using human duodenal juice (HDJ) or a commercial enzyme preparation consisting of porcine pancreatin and bile (PB). The lipolytic effect was determined by measuring the release of fatty acids (FA) using solid-phase extraction and GC-flame ionisation detection, withdrawing samples every 20 min during digestion. The amount of FA released indicated that a plateau was reached after 80 min with approximately similar amounts of FA detected using both HDJ and PB (379 (sd 18) and 352 (sd 23) mg/g SO, respectively). However, the release of 18 : 2, EPA (20 : 5) and DHA (22 : 6) was significantly different during in vitro digestion. At 80 min, HDJ and PB released 43 and 33% of 18 : 2, 14 and 9% of EPA and 11 and 9% of DHA, respectively. Both enzyme preparations released approximately the same amounts of the other FA analysed. The effect of the addition of bile salts (BS) was significantly different in the two enzyme systems, where porcine pancreatin highly responded to the increase in BS concentration, in contrast to HDJ.

Metabolism of the polycyclic musk galaxolide and its interference with endogenous and xenobiotic metabolizing enzymes in the European sea bass (Dicentrarchus labrax).

This study investigates the metabolism and mode of action of galaxolide (HHCB) in the European sea bass -Dicentrarchus labrax- following a single intraperitoneal injection of 50 mg HHCB/kg body weight. In addition, a group of fish was injected with 50 mg/kg of ketoconazole (KCZ), a fungicide that is known to interfere with different Cyp isoenzymes. HHCB was actively metabolised by sea bass and acted as a weak inhibitor of the synthesis of oxyandrogens in gonads of male fish. Both, HHCB and a hydroxylated metabolite were detected in bile. The fungicide ketoconazole was a strong inhibitor of Cyp11ß and Cyp3a-catalyzed activities. The work contributes to the better understanding of the impact of synthetic musks on fish and proposes the determination of HHCB and/or its hydroxylated metabolite in bile as a tool to assess environmental exposure in wild fish.

Identification of metabolites of ganoderic acid D by ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry.

Ganoderic acid D (GD) is the major active triterpenoid in Ganoderma lucidum, a medicinal fungus used daily. However, the metabolic fate of GD remains unknown. To know whether GD is extensively metabolized, we first investigated the metabolism of GD in vitro and in vivo. The metabolic profiles of the bile samples obtained from rats in vivo were almost the same as those obtained in vitro. Using ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry, a total of 25 metabolites were identified from the bile sample. Few metabolites were found in the urine samples. These results indicated that biliary rather than renal clearance was the major route of excretion. The major metabolites were identified by comparison with the standard reference compounds. Metabolites at low concentrations were identified by interpreting the mass spectra. Both phase I and phase II metabolites were observed. The metabolic transformation included reduction, monohydroxylation, dihydroxylation, trihydroxylation, oxidation, desaturation, sulfation, and glucuronidation. The main metabolic soft spots in the chemical structure of GD were the 3-carbonyl group, angular methyl groups, the 7-hydroxy group, and the 26-carboxylic acid moiety. Overall, this study gives us an insight into the metabolism of GD, an active oxygenated tetracyclic triterpenoid.

Effects of clonidine and idazoxan on tetrathiomolybdate-induced copper and lysosomal enzyme excretion into sheep bile.

This study investigated the effects of intravenous (IV) administration of tetrathiomolybdate (TTM), and α(2)-adrenergic agonist clonidine (CLO) and α(2)-antagonist idazoxan (IDA), alone or in combination with TTM, on sheep fed low (LCu) and high (HCu) copper diets. Effects on bile flow, biliary Cu concentration and excretion, plasma Cu concentration, and lysosomal enzyme ß-glucuronidase (ß-GLU) activity in bile and plasma were determined. Tetrathiomolybdate alone or with CLO or IDA significantly enhanced biliary Cu excretion most likely by removing Cu from hepatocyte lysosomes as evidenced by a significant increase in ß-GLU enzyme activity in bile. A significant increase in plasma ß-GLU concentration occurred only in sheep treated with CLO in combination with TTM. Because of the lytic nature of the lysosomal enzymes, caution is advocated in use of drugs, especially α(2)-adrenergic agonists, to further enhance TTM-induced biliary Cu excretion in the treatment of chronic Cu poisoning in sheep.

Clinical characteristics of liver fibrosis in patients with choledochal cysts.

PURPOSE: The aim of the study was to identify the clinical characteristics and outcome of patients with liver fibrosis in choledochal cyst (CC). METHODS: Forty patients with CC who underwent liver biopsy were included. Liver fibrosis was classified as follows: grade 0, no fibrosis; grade 1, mild fibrosis localized in the portal area; grade 2, moderate fibrosis with occasional bridging; and grade 3, severe fibrosis with diffuse bridging. RESULTS: Fourteen patients (35%) had liver fibrosis. Patients in the fibrosis group were significantly younger (1.2 vs 2.7 years) and had higher total bilirubin (5.3 vs 2.6 mg/dL). Severity of liver fibrosis was inversely correlated with age (P = .044). Amylase and lipase in bile were significantly lower in the fibrosis group (amylase, 531 vs 15,000 U/L; lipase, 783 vs 23,100 U/L). Postoperative serum analysis demonstrated no differences between the two groups. Most patients in both groups had normal aspartate aminotransferase, alanine aminotransferase, total bilirubin, and γ-glutamyl transpeptidase regardless of severity of fibrosis. Postoperative biliary complication or cholangiocarcinoma was not found in the fibrosis group. CONCLUSIONS: Our data suggest that liver fibrosis is mainly influenced by obstructive cholangiopathy rather than refluxed pancreatic secretion. Prognosis of patients with CC and liver fibrosis was as good as that of patients without fibrosis.

Measurement of carbonic anhydrase isozyme VI (CA-VI) in swine sera, colostrums, saliva, bile, seminal plasma and tissues.

Swine secretory carbonic anhydrase VI (CA-VI) was purified from swine saliva and an antibody to CA-VI was generated. A specific and sensitive enzyme-linked immunosorbent assay (ELISA) has been developed for the measurement of swine CA-VI. The assay can detect as little as 5 ng/mL of swine CA-VI. Typical standard curves were determined for a range of CA-VI solutions (7.8 to 500 ng/mL). The coefficients of variation for these solutions were less than 5%. When 500, 250 or 100 ng/mL of swine CA-VI was added to swine sera, the recoveries were 102.0%, 109.7% and 100.2%, respectively. The concentrations of CA-VI in the saliva (26.2 ± 30.4 µg/mL), sera (3.3 ± 4.9 ng/mL), bile (153.0 ± 114.0 ng/mL), seminal plasma (124.0 ± 39.0 ng/mL) and parotid gland (441.3 ± 90.0 µg/g wet tissue), submaxillary gland (88.1 ± 124.4 µg/g wet tissue), sublingual gland (58.6 ± 24.6 µg/g wet tissue) and gallbladder (2.4 ± 1.3 µg/1g wet tissue) were determined by ELISA. The concentration of CA-VI in colostrum was 163.3 ± 101.4 ng/mL and did not decrease within 10 days following parturition. An immunohistochemical reaction to anti-CA-VI antiserum was observed in the columnar epithelial cells lining the gallbladder. These data suggest that secretory CA-VI plays various roles in pH regulation and the maintenance of ion and fluid balance.

The impact of pharmacogenetics of metabolic enzymes and transporters on the pharmacokinetics of telmisartan in healthy volunteers.

OBJECTIVE: Telmisartan is mainly taken up into the liver by organic anion transporting polypeptide (OATP) 1B3, conjugated with glucuronate, and excreted into the bile. We investigated the relationship between genotypes of metabolizing enzymes and transporters and pharmacokinetics of telmisartan in clinical study. We also checked which enzymes are responsible for telmisartan glucuronidation. MATERIALS AND METHODS: We collected blood samples from 57 healthy volunteers who had participated in a clinical trial of telmisartan and examined the relationship between 14 mutations in six transporters/metabolic enzymes and pharmacokinetics of telmisartan. We also performed an in-vitro glucuronidation assay with recombinant uridine 5'-diphospho-glucuronosyltransferases isoforms and human liver microsomes. RESULTS: In the clinical study, area under the plasma concentration-time curve value from time zero to infinity, of telmisartan in heterozygotes of SLCO1B3 (encoding protein: OATP1B3) rs11045585 tended to be larger than that in homozygotes of wild-type alleles. Unexpectedly, 19 heterozygotes of UGT1A1*28, whose function was decreased, significantly increased its oral clearance compared with homozygotes of UGT1A1*1 alleles (1090±690 vs. 620±430 ml/min/body). Metabolic clearance of telmisartan in human liver microsomes obtained from individuals with UGT1A1*28/*28 was higher compared with that of UGT1A1*1/*1 (168±33 vs. 93.3±27.3 µl/min/mg protein). Although telmisartan was metabolized by multiple UGT isoforms, in-vitro experiments revealed that UGT1A3 was estimated to be predominantly involved in telmisartan glucuronidation in human hepatocytes. CONCLUSION: UGT1A1*28 was thought to enhance the protein expression of UGT1A3 as reported most recently (Riedmaier et al. Clin Pharmacol Ther 2010; 87:65-73) and thereby increase glucuronidation activity of telmisartan and decrease the plasma concentration of telmisartan.

Role of pressure and pancreatic reflux in the aetiology of choledochal malformation.

BACKGROUND: The aetiology of choledochal malformation is not known. Babbitt's hypothesis remains a popular concept, and assumes that activated pancreatic juice refluxes through the common pancreatobiliary channel causing mural damage and subsequent biliary dilatation. This hypothesis was tested clinically by evaluating the relationship between epithelial histology, choledochal pressure and degree of pancreatic reflux. METHODS: Children with choledochal malformation (cystic, type 1c; fusiform, type 1f; both intrahepatic and extrahepatic dilatation, type 4) operated on between January 1999 and October 2009 were identified. Where practical, choledochal pressure was measured on entry to the abdominal cavity, by puncture of the common bile duct, and bile was sampled for amylase content. Archival bile duct sections were scored using a semiquantitative epithelial lining/mural score (ELMS). RESULTS: A total of 90 children with choledochal malformations were operated on during the study interval. Histology was available for 73 children (median age 2·9 (interquartile range 1·3-7·9) years), 29 with type 1c, 31 with type 1f and 13 with type 4 malformations. There was a significant stepwise increase in pressure with choledochal morphology (median pressure 13, 17 and 20 mmHg for types 1f, 1c and 4 respectively; P = 0·037). There was an inverse relationship between choledochal pressure and bile amylase activity (r(s) = - 0·45, P < 0·001). High ELMS values were associated with higher choledochal pressure (P = 0·057) and low bile amylase activity (P = 0·002). CONCLUSION: High choledochal pressure (not bile amylase) was associated with more severe histopathological changes and choledochal morphology. These findings suggest that distal bile duct obstruction (and therefore high intraluminal pressure) contributes more to the key features of choledochal malformation than does pancreatic reflux.

Adherent properties and macrophage activation ability of 3 strains of lactic acid bacteria.

Lactic acid bacteria (LAB) must possess probiotic properties in order to be beneficial to humans and animals. The adherent properties, the acid and bile tolerance as well as the macrophage activation ability of isolated LAB strains were investigated in this study. The adhesion was analyzed following heat, acid, trypsin, and sodium periodate treatments. Production of the cytokine tumor necrosis factor-α (TNF-α) by RAW 264.7 macrophages was also measured after stimulation with heat-killed LAB strains. The viable strains of Lactobacillus fermentum AF7, L. acidophilus GG5, and L. plantarum BB9 were able to tightly adhere to the intestinal Caco-2 cells. In addition, the GG5 strain was not affected by heating, acid, trypsin, or sodium periodate treatment. However, the adhesion of strains AF7 and BB9 was reduced significantly by heating and trypsin treatment. This result suggested the GG5 and AF7 or BB9 strains had different cell-surface adherent factors. TNF-α production by the RAW 264.7 macrophages was induced significantly following stimulation with heat-killed LAB at 10(8) CFU/mL in a dose-dependent fashion. In addition, macrophage activity was similar whether the treatment consisted of live probiotics, or probiotics treated with heat, acid, or trypsin. However, the activity was reduced after treating with sonication. These in vitro results showed that the LAB studied possess probiotic characteristics, such as acid or bile tolerance, adherent capability, and immune activation, and may suggest that these LAB strains retain their probiotic activity as they pass through the gastrointestinal tract.

Intrahepatic duct dilatation in type 4 choledochal malformation: pressure-related, postoperative resolution.

BACKGROUND: Type 4 choledochal malformations (CMs) may be defined as those with both intrahepatic and extrahepatic bile duct dilatation. The aims of this study were to investigate possible causes of intrahepatic duct (IHD) dilatation in CM and to define the effect of surgery over time. METHODS: This study was a single-center retrospective review of a database of all children with CM undergoing surgery (excision of extrahepatic bile duct dilatation and hepaticojejunostomy) and identified as type 4 (on imaging and at surgery). Data included intraoperative choledochal pressure measurements and biliary amylase content and were expressed as median (interquartile range [IQR]). All comparisons used nonparametric statistical tests. P ≤.05 was regarded as significant. RESULTS: Twenty children were identified as type 4 CM (age, 4.3 years; range, 2.7-10.4 years) with preoperative IHD dilatation (right duct: diameter [range], 8.5 [4.5-14] mm; left: 8 [4-14.5] mm). Median intraoperative choledochal pressure was 17 (8-27) mm Hg (normal, <5 mm Hg), and intraoperative bile amylase was 3647 (range, 500-58,000) IU/L (normal, <100 IU/L). Preoperative IHD diameter correlated with choledochal pressure (right: r(s)=0.46, P = .03; left: r(s)=0.34, P = .07) but not with biliary amylase (P = .28 and P = .39, respectively). At 1 year postsurgery, median (range) IHD diameter had decreased to 1 (1-2.5) mm for right duct (P = .0002) and 1.5 (1-3) mm for left duct (P = .0006) and remained stable for up to a 10-year follow-up. CONCLUSION: Our data suggest that IHD dilatation is related to sustained increased intrabiliary pressure rather than any intrinsic intrahepatic CM. Effective surgery invariably reduces measured IHD toward normal values.

Reflujo pancreáticobiliar en pacientes con colelitiasis y sin colelitiasis: estudio comparativo / Pancreaticobiliary reflux in patients with cholelithiasis and without cholelithiasis: a comparative study

Background. Pancreaticobiliary reflux is a pathologic phenomenon occurring in patients with gallstones. However, the occurrence of pancreaticobiliary reflux has not been studied in patients without gallstones. The objective of this study was to measure the bile levels of amylase and lipase in patients without gallstones submitted to cholecystectomy as part of another surgical procedure, and to compare these values with patients submitted to cholecystectomy for gallstone disease. Patients and Methods. A prospective observational comparative study was designed. A sample of 136 consecutive patients was included. Amylase and lipase levels were measured in bile. Normal serum amylase levels at our institution are 28-100 U/L and for lipase are 13-60 U/L. There are no established normal levels for pancreatic enzymes in bile. However, we considered elevated the bile amylase and lipase levels whenever they were higher than normal plasma levels. Results. One-hundred three patients (76 percent) had gallstones and 33 (24 percent) liad healthy gallbladders without gallstones. According to normal plasma levels for amylase and lipase, these enzymes in bile were elevated in 83.5 percent patients with gallstones, compared to elevated levels of amylase in 6 percent patients and lipase in 3 percent patients without gallstones. Conclusions. Pancreaticobiliary reflux is a common phenomenon in patients with gallstones and occurs sporadically in patients without gallstones.

Introducción. El reflujo pancreáticobiliar es un fenómeno patológico que ocurre en pacientes con colelitiasis. La ocurrencia de este fenómeno no ha sido estudiada en pacientes sin colelitiasis. El presente estudio tiene por objetivo medir los niveles de amilasa y lipasa en la bilis de pacientes sin colelitiasis, colecistectomizados como parte de otro procedimiento quirúrgico y comparar estos valores con pacientes colecistectomizados por colelitiasis. Pacientes y Métodos. Se diseñó un estudio observacional y comparativo. Una muestra de 136 pacientes consecutivos fue incluida. Se midieron los niveles de amilasa y lipasa en la bilis. En nuestra institución los valores normales para amilasa son 28-100 U/L y para lipasa 13-60 U/L. No se han establecido valores normales de enzimas pancreáticas en la bilis. Para efectos del presente estudio, se consideró como elevados los niveles biliares de amilasa y lipasa cuando fueron mayores a los valores plasmáticos normales. Resultados. 103 pacientes (76 por ciento) tenían colelitiasis y 33 (24 por ciento) tenían vesículas normales sin cálculos. De acuerdo a los valores plasmáticos normales de amilasa y lipasa, estas enzimas se encontraron elevadas en 83,5 por ciento de los pacientes con colelitiasis comparados con valores elevados de amilasa en 6 por ciento en pacientes sin colelitiasis y de lipasa en 3 por ciento de estos pacientes. Conclusiones. El reflujo pancreaticobiliar es un fenómeno común en pacientes con colelitiasis y ocurre esporádicamente en pacientes sin colelitiasis.