Liver Amylase Research Articles

Apparent digestibility coefficients of carbohydrates and enzymes activity in major carps in response to Chromium Chloride Hexahydrate

This study investigated the effect of carbohydrate-based corn diets with chromium chloride hexahydrate supplementation on nutrient digestibility and enzyme activity in major carps (Cirrhinus mrigala, Catla catla, and Labeo rohita) for a period of 90 days. One control (T1) and five experimental diets (T2, T3, T4, T5, T6) were prepared by using gelatinized and non-gelatinized corn with the combination of various levels (0.0, 0.6, and 0.8 mg/kg) of CrCl3H12O6. After acclimatization with a control diet (T1) (32% CP), twenty fingerlings of these species were randomly stocked in eighteen digestibility tanks, each with one replicate. At the end of the trial, results showed that gelatinized corn with chromium supplementation significantly enhanced (P<0.05) digestibility of nutrients and enzyme activities (amylase and protease) in liver and gut in all treated groups. Crude protein (76.05±0.54), crude lipid (84.53±0.46), carbohydrate (69.15±2.19), and gross energy (76.92±0.74) showed the highest apparent digestibility in Labeo rohita as compared to Catla catla and Cirrhinus mrigala. While highest amylase (liver:3.27±0.126 U/ml and gut: 1.29±0.045 U/ml) and protease activity (liver:1.22±0.116 U/ml and gut 3.42±0.087 U/ml) was recorded in Catla catla subsequently followed by Labeo rohita and Cirrhinus mrigala. Among the treatments, highest amylase activity was observed in the corn gelatinized group at 0.6 mg/kg chromium level, while maximum protease activity was observed at 0.8 mg/kg of chromium level (gelatinized group). Comparison among gelatinized and non-gelatinized corn diets suggested that gelatinization in fish feed performed best as a carbohydrate energy source with chromium supplementation at an appropriate level (0.6 mg/kg) for three major carps as compared to non-gelatinized corn.

Alterations in biochemical parameters of fish species under choline administration directly into the pond water in a semi-intensive fish farming system: A comparative study

The present study deals with the impact of choline chloride supplementation, applied directly into the pond water in a semi-intensive farm culture system for 90 d on two Indian Major Carps (IMCs) (Catla catla, Labeo rohita) and two air-breathing teleostean fishes (Anabas testudineus, Clarias batracus).The experiment was conducted in two seasons (breeding and dry) under choline (treated) and non-choline (control) exposure. Biochemical parameters, viz., PRO (total protein) content gained maximum significantly (p< 0.01) in treated-breeding with the lowest in control-dry in intestine, liver, muscle, brain. IMCs showed significant (p< 0.01) increased amylase activity in the liver and intestine in treated-dry and treatment-breeding conditions, while liver amylase activity in both the seasons increased manifolds significantly (p< 0.01) in air-breathing fishes, showing a sharp decline only in the intestine. Protease and lipase activity in the intestine and liver of choline-fed fishes disclosed declining trend in the breeding season significantly (p< 0.01), but treated (choline-fed) IMCs under dry revealed the significant (p< 0.01) increasing and decreasing trend of protease and lipase activity respectively; the reverse trend at dry season was found in air-breathing fishes under choline-fed condition. ALP (alkaline phosphatase) in intestine indicated significant (p< 0.01) lowest activity in treatment-breeding, highest in treatment-dry. This study explored the higher activity towards total protein generation, especially muscle; higher carbohydrate digestion; elevated lipid digestion in choline-fed fishes. So, the fishes from this farm-based pisciculture system are healthy and its yield can be prescribed as a rich source of quality fish food for human health.

The positive effects of single or conjoint administration of lactic acid bacteria on Channa argus: Digestive enzyme activity, antioxidant capacity, intestinal microbiota and morphology

In the present study, we aimed to evaluate the single or conjoint effects of Lactococcus lactis (L. lactis) L19 and Enterococcus faecalis (E. faecalis) W24 isolated from the intestine of Channa argus (C. argus) on digestive enzyme activity, antioxidant capacity, intestinal microbiota and morphology of C. argus (initial weight, 9.50 ± 0.03 g). The fish were fed for 56 days with a basal diet (CK) or diets supplemented with 1.0 × 108 CFU/g of L. lactis (L19), E. faecalis (W24), L. lactis + E. faecalis (L + W). Results indicated that these three lactic acid bacteria (LAB) supplementary diets especially the L19 produced significant improvement in the digestive enzyme activity (protease, amylase and lipase) in liver, stomach and intestine compared with control (P < .05). LAB supplementations were enhanced the liver and intestine antioxidant status (SOD, CAT, GSH-Px, T-AOC and MDA) in comparison to the control (P < .05). Serum biochemical parameters (ALT and AST) of fish fed with LAB supplemented diets showed significantly decreased compared with control (P < .05). The intestinal morphology showed protection in fish fed both L19 or/and W24. High-throughput sequencing revealed L19 supplementation can effectively increase the generation of probiotic bacteria and decrease the abundance of aquatic pathogens. Therefore, supplementation of L. lactis L19 showed more effective protection than E. faecalis W24 or the mixture of the two for promoting digestive enzyme activity, antioxidant capacity, intestinal microbiota and morphology of C. argus.

Efficacy of Thiol Disulfide Homeostasis and Ischemia Modified Albumin Values in Estimating the Degree of Difficulty for Laparoscopic Cholecystectomy.

The aim of this study was to investigate the efficacy of thiol disulfide homeostasis and Ischemia Modified Albumin (IMA) values in predicting the technical difficulties that might be encountered during laparoscopic cholecystectomy. The study included 65 patients who underwent laparoscopic cholecystectomy due to cholelithiasis at the General Surgery Clinic of Ankara Numune Training and Research Hospital. All patients' demographic data, previous history of cholecystitis, a history of chronic illness, preoperative white blood count (WBC), liver function tests (AST, ALT), amylase and lipase levels, intra-operative adhesion score, the ultrasonographic appearance of gall bladder, duration of hospital stay, duration of operation, thiol disulfide and IMA values were evaluated. Native thiol and total thiol averages were higher in patients without a history of cholecystitis, and on the other hand, disulfide, disulfide/native thiol rate, disulfide/total thiol rate, native thiol/total thiol rate and IMA averages were higher in patients with a history of cholecystitis. While there was a statistically significant negative correlation between native and total thiol values and age, duration of surgery and duration of hospital stay; IMA, disulfide, disulfide/Total thiol, Native/Total thiol and disulfide/Native thiol rates were higher in older patients with a longer duration of surgery and hospital stay. In addition, preoperative IMA, disulfide, disulfide/Total thiol, Native/Total thiol and disulfide/Native thiol were observed to increase as the degree of intraoperative pericholecystic adhesion increased. We believe that the evaluation of thiol disulfide homeostasis and IMA parameters prior to laparoscopic cholecystectomy can be used as an effective method for predicting intraoperative difficulties.

Monitoring of growth, digestive enzyme activity, immune response and water quality parameters of Golden crucian carp (Carassius auratus) in zero-water exchange tanks of biofloc systems

A 56-day feeding trial was implemented to evaluate the effects of promoted biofloc on the water quality, growth performance, digestive enzymes activity, immune-related genes and antioxidant status of golden crucian carp in negligible exchange culture tanks. Four handred fifty healthy golden crucian carp juveniles (5.03 ± 0.22 g) were randomly assigned among 15 tanks. Five C:N ratios in triplicate tanks were tried: C/N = 5.48:1 with a commercial diet (Control), C/N = 10:1, C/N = 15:1, C/N = 20:1 and C/N = 25:1. Anhydrous glucose (99.97 %) was added to the commercial diet to maintain elevated carbon ratio in the three treatments. Uninterrupted 24 -h aeration was supplied during the test by using an air-stone connected to an air pump to develop the biofloc in the water column. The results demonstrated that BFT significantly reduced ammonia nitrogen (TAN), nitrite (NO2−-N), total phosphorus (TP) concentration and water transparency (Secchi depth) in C/N 15–25 than control (p < 0.05), wheras the chemical oxygen demand (COD) and nitrate nitrogen (NO3−-N) concentration gradually increased over time. Apart from, weight gain (WG), specific growth rate (SGR), and protein efficiency ratio (PER) were considerately elevated in C/N 20 and C/N 25 than control (p < 0.05).This was coincidental with the higher activities for protease, lipase and amylase in liver. Significant increases in lysozyme (LSZ), acid phosphatase (ACP) and alkaline phosphatase (AKP) activities of golden crucian carp were found in the C/N 20 group than control (p < 0.05), and regulating the expression of immune-related genes including IL-1β, TNF-α and HSP70 after a 56-day feeding trial. Comparing the antioxidant capacity of golden crucian carp from the five experimental groups, the activity of superoxide dismutase (SOD), total antioxidant capacity (T-AOC), catalase (CAT) activity and glutathione peroxidase (GSH-PX) activity of juveniles in the C/N 20 and C/N 25 groups were significant higher (p < 0.05), and the content of malondialdehyde (MDA) was much lower (p < 0.05) than in the control under this experiment condition. Overall, this findings suggest that BFT not only can enhance golden crucian carp growth performance, elevate digestive enzymes activity, and strengthen antioxidant status but also upgrade immune response.

Determination of Some Human Enzymes Activities in Presence of Titanium Oxide Nanoparticles

Titanium oxide nanoparticles (TiO2NPs) are the most common particles that have industrial, commercial, medial and antibacterial applications. On the other hand, they have toxic properties within some concentrations that show the ability to inhibit some human enzymes. In this study a diluted solutions of TiO2NPs were prepared by weighting(0.08 gm)with deionized water as(10−1, 10−2, 10−3, 10−4, 10−5) with low conductivity to avoid interaction with other ions. Our study aimed to estimate the effect of different concentration of TiO2NPs on liver enzymesaspartate transaminase (GOT), alanine transaminase (GPT), alkaline phosphatase(ALP) and Amylase in human serum by using a healthy human serum with a mixture of enzyme solution in absence and presence of diluted solution of TiO2NPs. The results showed that different TiO2NPs concentrations used had inhibitory effect on all the enzymes.

An uncommon etiology of nausea and vomiting: paraesophageal hernia

A 62 year old female on omeprazole with a history of hiatal hernia, gastroesophageal reflux disease (GERD), and esophagitis presented with acute onset of nausea and vomiting. She vomited approximately 20 times, quantified as half a cup mixed with dark red blood. Initially, the emesis was all recently ingested meals. The emesis became bloody after 24 hours of vomiting, leading to her presentation in the ER. She continued to feel nauseous with belching in the ER. She denied any abdominal pain, diarrhea, constipation, chest pain, odynophagia or shortness of breath. She had no melena, hematochezia, fevers, chills or weight loss. She denied any NSAID use or recent international travel. She had no family history of gastrointestinal malignancy and no history of tobacco use. Her physical examination and vital signs were unremarkable. Laboratory data was significant for white cell count of 18.78 K/µL, hemoglobin of 15.7 G/DL, hematocrit 46.3% (hemoglobin & hematocrit at baseline), mean corpuscular volume 88FL, platelet count 65 K/µL and INR was 1.0. Her chemistry, liver function test, lipase and amylase were within normal limits. Computed tomography(CT) scan of the chest was done.

Expression of liver alpha-amylase in obese mouse hepatocytes.

The aim of this study is to demonstrate the relation between the expression of liver alpha-amylase and obesity. Alpha-amylase catalyses the hydrolysis of 1, 4-alpha-glucosidic linkages in polysaccharides and has three main subtypes, including: salivary, pancreatic, and hepatic. Hepatic alpha-amylase is involved in glycogen metabolism, and has a role in obesity and its management. In this study, we aimed to analyze the expression of liver alpha-amylase in overweight and obese mouse. In this study, NMRI male mice were randomly divided into two groups. The sample group (obese) took a high-fat and carbohydrate diet, while the control group (normal) took a laboratory pellet chow for eight weeks. During this period, their weight was measured. After eight weeks, liver hepatocytes were isolated using an enzymatic digestion method. Immunocytochemistry (ICC) and flow cytometry analysis were performed to measure alpha amylase protein expression in mouse liver hepatocyte cells. A significant difference in the body weight was observed between the two groups (p<0.05). The qualitative protein expression of liver alpha-amylase was found to be higher in the obese group in both tests (immunocytochemistry and flow cytometry). Animals from the test group presented higher alpha-amylase expression, which suggests that this hepatic protein may constitute a potential indicator of susceptibility for fat tissue accumulation and obesity. The present data demonstrates an increased expression of liver amylase in obese mice. These results suggest that liver amylase secretion might be useful for predicting susceptibility to obesity induced by consumption of a high-fat and carbohydrate diet.

Effects of dietary supplementation with green tea waste on growth, digestive enzyme and lipid metabolism of juvenile hybrid tilapia, Oreochromis niloticus×O. aureus.

An 8-week feeding trial was conducted to evaluate the effects of dietary supplementation with green tea waste (GTW) on growth, digestive enzyme and lipid metabolism of juvenile hybrid tilapia, Oreochromis niloticus×O. aureus. The fish (initial mean body weight, 12.63±0.75g) were fed five experimental diets that included 0 (control), 0.8, 1.6, 3.2 or 6.4% of GTW in triplicate aquaria, twice daily. Growth performance, plasma metabolites content and liver and intestine digestive enzyme activities were determined. Fish accepted well all experimental diets during the trial, and no mortality was observed. The weight gain increased (P<0.05) with the increase in GTW inclusion level up to 1.6%, after which it decreased, but no significant differences between the control and high level (3.2 or 6.4% of GTW) groups were observed. Moreover, fish fed on diets containing 0.8 and 1.6% GTW had lower feed conversion ratio (FCR, 1.75 and 1.73, respectively) and had better protein deposition (higher protein efficiency ratio, PER, 1.73 and 1.71, respectively), compared to other treatments. No differences among groups were observed in whole body and dorsal muscle composition with the exception of lipid content which was lower in fish fed 6.4% GTW diets, compared to other treatments. Lipase activities in liver or intestine were higher in fish fed GTW-supplemented diets with the exception of intestine lipase activities, which was unaffected, compared to the control. Similarly, liver lipoprotein lipase activities were also increased in fish fed diets supplemented a medium dose of GTW (1.6 or 3.2%), compared to other treatments. However, intestine amylase activities were decreased in fish fed diets containing a high dose of GTW (3.2 and 6.4%); while the liver amylase activities were unaffected by the GTW supplementation. Blood chemistry parameters were affected by GTW inclusion, except the values of triglycerides, which was unaffected. The values of total cholesterol, HDL cholesterol and LDL cholesterol increased with increasing GTW inclusion level up to 3.2%, after which the values decreased. These results indicate that diets supplemented with appropriate concentration of GTW (from 0.8 to 1.6%) may potentially serve as an effective functional food and additive for tilapia to improve growth performance, digestion efficacy and fat metabolism.

Dietary lipid requirements of larval genetically improved farmed tilapia,Oreochromis niloticus(L.), and effects on growth performance, expression of digestive enzyme genes, and immune response

The present study was performed to investigate the effects of dietary lipid level on growth performance, antioxidant defense, and mRNA levels of digestive enzymes and immune genes in larval genetically improved farmed tilapia (Oreochromis niloticus). The results show that larvae fed lipid levels of 4–11.5% had greater weight gain (WG), feed efficiency, protein efficiency ratio and survival than larvae fed the other dietary lipid levels (P < 0.05). No difference whole-body crude protein, ash or moisture contents were found between treatments (P > 0.05) but lipid contents increased as dietary lipid level was increased. The 8% and 11.5% dietary lipid levels had stimulatory effects on trypsin mRNA level compared with those of larvae fed the 15% and 18.5% lipid diets. Intestinal and liver amylase mRNA levels decreased in larvae fed the 18.5% lipid diet, following the decrease in dietary starch. Analysis of the secondary curve equation for WG indicated that the optimal dietary lipid level in larval tilapia was 8.56%. In addition, the 4–11.5% lipid levels improved superoxide dismutase and glutathione peroxidase activities and increased mRNA levels of heat shock protein70, hepcidin antimicrobial peptide-1, and C-type lysozyme in the liver of larvae (P < 0.05). A 192-h Streptococcus iniae challenge study was conducted after the 35-day feeding trial, and the results showed that post-challenge mortality of larvae fed the 11.5% lipid level was the lowest and enhanced the immune activities.

Alterations in Digestive Enzymes of Three Freshwater Teleostean Fishes by Almix Herbicide: A Comparative Study

Three freshwater teleostean fishes viz., Anabas testudineus (Bloch), Heteropneustes fossilis (Bloch) and Oreochromis niloticus (Linnaeus) were exposed to almix (66.67 mg/l) herbicide for 30 days to investigate the activity of digestive enzymes (amylase, lipase and protease) in stomach, intestine and liver. Amylase activity showed significantly high (p A. testudineus > H. fossilis.

Effects of light intensity on husbandry parameters, digestive enzymes and whole-body composition of juvenileEpinephelus coioidesreared in artificial sea water

To provide a specific lighting system to aquaculture, a completely randomized design with five light treatments with three replicates was used to investigate effects of light intensity on juvenile Epinephelus coioides reared in artificial sea water. The weight gain rate (WG, %), specific growth rate (SGR % day−1), protein efficiency ratio (PER) and survival rate (S, %) were significantly higher at the 320–1150-lx treatments (P ≤ 0.05) compared with the other treatments, but the highest food conversion ratio (FCR) was at the 0-lx treatment and the lowest at 600–1150 lx. Total protease, amylase and lipase activities in liver, stomach and intestine were significantly higher at 320–1150-lx conditions and significantly lower at 0 lx (P ≤ 0.05) than in the other treatments. The highest contents of crude protein and crude lipid were observed at 600–1150 lx and 320–550 lx respectively. However, both moisture and ash were highest at 0 lx and lowest at 600–1150 lx. The results indicated that light intensity could significantly affect husbandry parameters, digestive enzymes and whole-body composition of juvenile E. coioides, with optimal light intensity being at 600–1150 lx.

Catastrophic antiphospholipid syndrome (Asherson's syndrome) presenting with a splenic rupture

A PREVIOUSLY HEALTHY 51-YEAR-OLD JAPANESE WOMAN was admitted to our hospital as an emergency case because of hemorrhagic shock owing to intraabdominal bleeding without any trauma. The patient was conscious, blood pressure was 56/38 mm Hg, pulse 90 beats/min, and body temperature was 36.8 C. Her abdomen was distended and diffusely tender. Rebound tenderness was noted, but the abdominal defense reflex was absent. Bowel sounds were silent and there was no flatus. The patient had a past history of 3 spontaneous abortions, but no history of laparotomy. Laboratory tests revealed a leukocyte count of 8,690/mm with 82% neutrophils and 4.5% lymphocytes, 6.6 g/dL hemoglobin, 19.3% hematocrit, and a platelet count of 11,000/mm. Coagulation tests revealed a prothrombin time of 33% (International Normalized Ratio, 2.0), activated partial thromboplastin time of 54.6 seconds, and antithrombin III of 58%. Total proteins were 4.0 mg/dL with albumin at 2.1 g/dL. The fasting blood glucose level, liver enzyme level, creatinine, uric acid, amylase, lipase, and electrolytes were within normal limits. A contrast-enhanced computed tomograph of the abdomen showed a massive collection of intraabdominal fluid, portal vein thrombosis with splenic infarction, and hypodense lesion at the periphery of the spleen which was compatible with hematoma (Fig, A). An emergency laparotomy was performed immediately after carrying out a red blood cell and

Biliary tract prosthesis combined with pancreatic duct stents in patients with periampullary carcinoma

Objective To evaluate the efficacy of biliary tract prosthesis and pancreatic duct stents for advanced periampullary carcinoma. Methods A total of 36 patients were diagnosed as advanced periampullary carcinoma pathologically or clinically, with strictures both in pancreatic and biliary ducts confirmed by imaging. Teflon stents were firstly implanted through endoscopy to the narrowed pancreatic ducts, expansible metal prosthesis were then implanted to the biliary tract. If failed, the metal stents were given through percutaneous transhepatic biliary drainage （ PTCD ） pathway. Serum levels of liver enzymes, amylase and clinical manifestations were observed before and after operation. Results Teflon stents were successfully implanted into pancreatic ducts in all patients. Metal prostheses into bile ducts were endoscopically implanted in 29 cases, and via PTCD in 7, including 2 cases of Billroth II gastrectomy. The levels of liver enzymes significantly decreased （P 〈 0. 01 ） after stents implacement. The levels of amylase （plasma and urine） and lipase increased in 15 cases, but they were corrected to normal levels after adequate treatments. Rates of abdominal pain relieving and diahhrea improvement were 82. 4% （28/34） and 88. 2% （ 15/17 ）, respectively. Conclusion The combined implacement of biliary tract prosthesis and pancreatic duet stents is safe and effective in relieving malignant obstruction in periampullary carcinoma. Key words: Biliary tract neoplasms; Malignant obstruction; Bile ducts ; Pancreatic duets; Stents

The Exposure Of &lt;i&gt;Heterobranchus bidorsalis&lt;/i&gt; Juveniles To Different Concentrations Of Bonny-Light Crude Oil And Their Effects On Amylase And Cretinine Kinase Activities

The effects of exposing Heterobranchus bidorsalis juveniles (14.08 ± 0.12 g) to diferent concentrations of Bonny-light crude oil (BLCO) on amylase and cretinine kinase activities were studied The exposure of the fsh to 1.00, 2.00, 400, 8.00 ml L-1 BLCO and a control for 4 days toxicity period indicated that the signiicant increases (P < 0.01) in the serum amylase (SRA) and the hepatic cytosolic amylase (HCA) activities in the fish were BLCO concentrations dependent. Reduced SRA and HCA activities noticed within the first 14 days o the recovery period implied thatthe removal o the oil polutant from the ambient water chemistry probably lowered the pressure on the blood serum and liver amylase enzyme to catalyse the metabolism o the ingesed carbohydrates. Signicant increases (P < 0.05) in the serum cretinine kinase (SRCK) and the hepatic cytosolic cretinine kinase (HCCK) activities in the fish also followed the pattern shown by the SRA and the HCA activities. The increased SRA, HCA, SRCK and HCCK activities in the blood serum and liver of the fish were indications o a shit in the carbohydrate metabolism due to crudeoil exposure. Keywords: Heterobranchus bidorsalis, Bonny-light crude oil, Serum, Cytosolic, Amylase, Cretinine kinase.Animal Research International Vol. 3 (3) 2006 pp. 516-520

Cholecystocolonic fistula: an unusual cause of large bowel obstruction

A 70-year-old woman was admitted to the emergency department with a 2-week history of pain in the upper abdomen, vomiting, fever and jaundice. She was also complaining of pale stool and dark urine. Subsequently she developed abdominal distension with colicky pain and absolute constipation. There was no history of previous biliary symptoms. On examination she was pale and tachycardic with a fever of 37.9°C. Abdominal examination revealed generalized abdominal distension with a tender mass at right upper quadrant and hyperactive bowel sounds on auscultation. Laboratory examination showed a white blood cell count of 15 600/mm3 (neutrophil 85%), bilirubin of 47mmol/litre, alkaline phosphatase of 220 U/litre with normal liver enzymes and amylase. Plain abdominal X-ray demonstrated pneumobilia and dilated small and large bowel loops suggestive of intestinal obstruction (Figure 1). An unprepared barium enema showed a large smooth filling defect in sigmoid colon with large bowel obstruction (Figure 2). A diagnosis of large bowel obstruction presumably caused by a gall-stone was made. Surgical treatment followed a period of physiological optimization. At laparotomy the abnormalities were large bowel obstruction caused by an impacted gall-stone in a section of sigmoid diverticular disease plus right upper quadrant mass. A sigmoid colectomy with removal of diverticular disease and the stone, then a primary anastomosis was performed. Exploration of right upper quadrant mass revealed an inflamed friable gall bladder with a fistula between its fundus and the transverse colon. A partial cholecystectomy was performed, the cystic duct closed in-situ and the defect in transverse colon was brought out as a loop colostomy. Her postoperative course was uneventful. Closure of colostomy was performed 3 months later and she made a full recovery.

Generation of glycogen- and albumin-producing hepatocyte-like cells from embryonic stem cells

We present a novel two-step protocol for the differentiation of embryonic stem (ES) cells into the hepatic lineage. Differentiated hepatocyte-like cells express genes and proteins characteristic for endodermal and hepatic cells and acquire a functional hepatic phenotype as demonstrated by albumin secretion and glycogen storage. During differentiation, alpha-fetoprotein, albumin, transthyretin, alpha-1-antitrypsin, cytochrome P450 subunits 2b9 and 2b13 and tyrosine aminotransferase transcripts are upregulated. Quantitative RT-PCR data revealed a fetal hepatic phenotype corresponding to day 13-14 of liver development. Terminally differentiated hepatocyte-like cells show a bi-nucleated, cuboidal morphology labeled by albumin, alpha-1-antitrypsin, liver amylase, dipeptidyl peptidase IV, c-met and cytokeratin 18. ES-derived intermediate cell types transiently and partially co-express nestin with albumin and alpha-fetoprotein, respectively, but not cytokeratin 19. This finding suggests an ES-derived potential hepatic progenitor cell type, which is partially nestin-, albumin- and alpha-fetoprotein-positive, but cytokeratin 19-negative.

Electrophoretically unique amylases in rat livers: phylogenic and ontogenic study on the mammalian liver.

Liver amylase activity in rodents was assayed with Blue Starch as substrate, and found to be higher than in humans or pigs. Based on the result of concanavalin A affinity chromatography, we found that the sugar moieties of amylase molecules increased in parallel with amylase activity in the tested mammals. However, the amounts of amylase proteins determined by Western bloting with anti-human salivary-type antibody as the probe, were similar to the levels in mammalian livers. Moreover, a similar expression of amylase mRNA was also detected in the mammalian livers by a reverse transcriptional-polymerase chain reaction using primers specific for the human salivary and/or pancreatic amylase complementary DNA (cDNA) sequences. The amylase was detected at the catalytic activity, protein molecule and mRNA levels in rat liver at all ages from fetus to adult. Salivary-type liver amylase activity increased up to one week after birth, and was maintained at the adult level thereafter. However, based on the results of the electrophoretic mobility test, livers with accelerated amylase activity, e.g., at 2-4 weeks after birth or during liver regeneration after partial hepatectomy, were also found to express an amylase electrophoretical identical to pancreatic-type amylase in addition to salivary-type activity. These results suggest that the liver may express an etopic amylase in a certain condition.

Human Interleukin 10 Gene Therapy Decreases the Severity and Mortality of Lethal Pancreatitis in Rats

Background. Studies have proven the validity of interleukin-10 (IL-10) in the treatment of experimental pancreatitis. Prophylactic human IL-10 (hIL-10) gene treatment attenuated the severity in cerulein models. Our research aims to study whether the therapeutic hIL-10 gene could decrease both severity and mortality in a lethal pancreatic model.Methods. Severe acute pancreatitis (SAP) was induced by sodium taurocholate. A plasmid–hIL-10 construct (pcDNA3–hIL-10) complexed with cationic liposomes was administered to SAP rats by a single intraperitoneal injection. Levels of hIL-10 in the pancreas, liver, and lungs were determined by ELISA kits. The severity of pancreatitis was assessed in terms of serum amylase, histology, and tissue tumor necrosis factor α (TNF-α). Mortality, observed for 7 days, was evaluated for gene therapy or control groups.Results. After hIL-10 gene therapy, hIL-10 levels in the pancreas, liver, and lungs increased significantly and the serum amylase, tissue TNF-α, and histological changes in pancreas, liver, and lungs decreased markedly. Therefore, mortality was significantly reduced in the hIL-10 gene therapy group, in which 70% of rats survived in the 7-day observation, while only 10% survived in untreated groups (P < 0.05).Conclusion. We found that liposome/hIL-10 gene therapy decreased severity and mortality in SAP, even carried out after SAP establishment, predicting a more convenient shift to clinical applications.

α-Amylase expressed in human liver is encoded by the AMY-2B gene identified in tumorous tissues

Background: An α-Amylase in human liver is detected with an anti-human salivary amylase antibody, but the enzyme activity is very low. We previously found that the rat liver contained an amylase which differed from the enzyme of mice. In this study, we characterized the human liver amylases biochemically and immunohistochermically. Methods and Results: Although the amylase activity of human liver was much lower than that of rat, protein moiety and sugar chains of the human amylase were identified as similar to the rat liver enzyme with an anti-human salivary amylase antibody and by concanavalin A (Con A) affinity chromatography. Liver amylases from human and rat were the same size, 50 kDa, on Western blot analysis and had the same isoelectric points. The cytoplasm of hepatocytes was moderately stained immunohistochemically with the anti-human salivary amylase antibody. Intrahepatic bile ducts were also stained weak-to-moderately. RT-PCR, with a specific primer for the consensus sequence of human amylases, amplified a single 474-bp product from the human liver total RNA. The PCR product was sequenced and referred to the homology. Thirteen bases in the 434-bp fragment of the human liver amylase differed from the corresponding region of the AMY-1 gene transcript and the deduced amino acid sequence differed at five residues. The human liver amylase cDNA sequence was identical to the corresponding cDNA of the AMY-2B, which was known to expressed in tumorous tissues. In situ hybridization revealed the expression of AMY-2B mRNA in non-tumorous human liver. Conclusions: The present results suggest the possibility that a novel amylase detected in tumorous tissues and encoded by the AMY-2B gene is a liver-specific amylase expressed in the human liver.