Ultrastructural changes in the rat exocrine pancreas after jejunoileal bypass

The influence of a 90% jejunoileal bypass on the rat exocrine pancreas was studied by morphometrical procedures. In sham-operated animals exocrine acinar cells accounted for 80.3% of the pancreas volume. These cells are composed of 9.9% nuclei, 8.4% mitochondria, 12.2% zymogen granules, 0.3% lipid droplets and 69.2% of a compartment ("ERGLS") composed of endoplasmic reticulum, ribosomes, Golgi areas, lysosomes and the cytoplasmic ground substance. Intestinal bypass did not change the volume density of exocrine cells nor that of nuclei in the cells during the three postoperative months. The means nuclear diameter was approximately the same in both groups. However, the volume density of secretory granules diminished by 50%. This was accompanied by a decrease in mean granular diameter, but not in their numerical density. The volume density of lipid droplets increased 10 fold, that of mitochondria increased slightly from the 15th postoperative day but significantly from the 45th day. The remaining cellular compartment composed of "ERGLS" was not modified by intestinal bypass. These findings suggest that a 90% jejunoileal bypass induces major changes in the composition of pancreatic acinar cells but not in their size.     

Leucine absorption after jejunoileal bypass in rats

Jejunal and ileal absorption of L-leucine has been studied in rats subjected to jejunoileal bypass and in sham-operated rats, for five minute periods, using a perfusion technique. Aminoacid concentrations were: 1, 2.5, 5, 10 and 25 mM. In some experiments methionine was added to determine simple diffusion. The ratio of the active/diffusive components of absorption were calculated at the different luminal aminoacid concentrations in both groups of rats, showing that this ratio was lower in control animals.     

Effect of jejunoileal bypass in the rat on the enteroinsular axis

The effect of jejunoileal bypass (JIB) on the enteroinsular axis was studied in vivo and in vitro in the rat. Glucose, insulin and GIP responses to oral glucose were compared in JIB and control rats. The effect of glucose and GIP on insulin release from the isolated perfused pancreas of the same animals was investigated to determine if JIB altered the sensitivity of the beta cell. Immunocytochemical studies of gut and pancreas were also carried out. Glucose, insulin and GIP responses to a glucose load were blunted after JIB, although basal GIP levels were elevated in these animals. The insulin response of the perfused JIB pancreas to GIP was 70% reduced from controls although the insulin response to glucose appeared normal. The size and area of JIB islets were unchanged from controls as was the distribution of insulin, glucagon, somatostatin and pancreatic polypeptide. GIP immunoreactive cells were present in all regions of the intestine including the JIB blind loop. This study confirms the findings of others that a relationship exists between reduced GIP and insulin response to oral glucose after JIB, and indicates that a decrease in sensitivity of the beta cell to GIP occurs following JIB that is not rapidly reversible. GIP secreted from blind loop mucosa may contribute to the high basal GIP found in JIB rats and may be causally connected to the fall in beta cell sensitivity.     

Disaccharidase activities in pregnant and lactating rats

The changes in intestinal disaccharidase activities in pregnancy and lactation could be explained as a result of an adaptation to the energy demands of each reproductive stage. To examine these changes, disaccharidase activities (lactase, sucrase, maltase, and trehalase) were measured in the jejunum and ileum of virgin, pregnant and lactating Wistar rats. Minor changes in disaccharidase activities were observed in pregnancy, whereas an increase of disaccharidase activities per small intestine length normalized to body mass was observed in lactation.     

Hepatic structure and function after modified jejunoileal bypass surgery for obesity.

The most serious adverse effect of standard intestinal bypass for obesity is the high incidence of hepatic dysfunction and death from hepatic failure. We therefore examined the long-term effects of a modified form of jejunoileal bypass (in which a greater continuous length of ileum is retained), on liver function in 120 patients. Substantial weight loss (119-0+/-SD 23-3 kg to 82-3+/-18-8 kg) occurred during the first nine months after surgery, accompanied by a significant rise in serum concentrations of bilirubin, alanine transferase, and alkaline phosphatase, and a significant reduction in albumin concentrations. Biochemical changes were unrelated to weight loss or halothane anaesthesia. After weight stabilisation liver function reverted to normal, and four years after bypass sulphobromophthalein retention and hepatic histology did not differ from those in obese controls. There were two postoperative deaths. Three other patients died during the period of rapid weight loss with severe hepatic steatosis. While transient mild impairment of liver function is common after modified jejunoileal bypass, clinically significant hepatic dysfunction is a rare and unexplained early complication.     

Disaccharidase activities in camel small intestine: biochemical investigations of maltase-glucoamylase activity

Disaccharidases (maltase, cellobiase, lactase, and sucrase), alpha-amylase, and glucoamylase in the camel small intestine were investigated to integrate the enzymatic digestion profile in camel. High activities were detected for maltase and glucoamylase, followed by moderate levels of sucrase and alpha-amylase. Very low activity levels were detected for lactase and cellobiase. Camel intestinal maltase-glucoamylase (MG) was purified by DEAE-Sepharose and Sephacryl S-200 columns. The molecular weight of camel small intestinal MG4 and MG6 were estimated to be 140,000 and 180,000 using Sephacryl S-200. These values were confirmed by SDS-PAGE, where the two enzymes migrated as single subunits. This study encompassed characterization of MGs from camel intestine. The Km values of MG4 and MG6 were estimated to be 13.3 mM and 20 mM maltose, respectively. Substrate specificity for MG4 and MG6 indicated that the two enzymes are maltase-glucoamylases because they catalysed the hydrolysis of maltose and starch with alpha-1,4 and alpha-1,6 glycosidic bonds, but not sucrose with alpha-1,2 glycosidic bond which was hydrolyzed by sucrase-isomaltase. Camel intestinal MG4 and MG6 had the same optimum pH at 7.0 and temperature optimum at 50 degrees C and 40 degrees C, respectively. The two enzymes were stable up to 50 degrees C and 40 degrees C, followed by strong decrease in activity at 60 degrees C and 50 degrees C, respectively. The effect of divalent cations on the activity of camel intestinal MG4 and MG6 was studied. All the examined divalent cations Ca(2+), Mn(2+), Mg(2+), Co(2+) and Fe(3+) had slight effects on the two enzymes except Hg(2+) which had a strong inhibitory effect. The effect of different inhibitors on MG4 and MG6 indicated that the two enzymes had a cysteine residue.     

Phosphatase activities in rat liver before and after birth

Synopsis Several phosphatase enzymes have been studied biochemically and cytochemically to ascertain whether there are ontogenic changes in level or location. Nucleoside monophosphatase (5-nucleotidase) and lysosomal acid phosphatase are low in foetal liver and, unlike glucose-6-phosphatase, are still quite low in neonatal liver. Bile canaliculi show strong staining for 5-nucleotidase in adult liver but not in foetal or neonatal liver. Nucleoside diand triphosphatase activities in foetal liver are already near half the adult level. The diphosphatase that is active towards UDP shows the same cytochemical locations in neonatal liver as in adult liver. Triphosphatase activity in foetal and neonatal liver is located largely in star-like cells, rather than in the bile canaliculi of parenchymal cells. Biochemical comparison of foetal, neonatal and adult liver has shown that inorganic pyrophosphatase (assayed without Mg²⁺) parallels glucose-6-phosphatase, but acid ribonuclease does not parallel acid phosphatase. In albino rats injected with thyroxine, glucose-6-phosphatase has shown a more marked increase in foetal liver than in adult liver, although the uptake of thyroxine seemed to be less. In hooded rats, foetal liver showed a negligible uptake of thyroxine and no rise in glucose-6-phosphatase.A. A. El-Aaser is on leave from the Faculty of Medicine, University of Cairo.     

Changes in lipid parameters after intestinal resection or bypass in the rat.

Intestinal resection, bypass and adaptative postoperative mechanisms developed as a consequence of that surgery, are considered good methods for improving knowledge of gastrointestinal physiology as well as possible effects that the intestine could have on the general metabolism. 50% jejunoileal bypass (BP), 50% proximal (PR) and distal (DR) intestinal resections were performed on rats to compare the influence of resected intestinal segments or bypassed loop localization could exert on different serum lipid parameters. One month after surgery significant increases in total serum cholesterol and cholesterol esters were found. There was no change in free cholesterol. A decrease in triglyceride was observed after distal and proximal resection but no changes after bypass. The cholesterol/phospholipid ratio was increased after resection and after bypass. It has been suggested that the changes in lipid metabolism produced after resections and bypass depend mainly on the loss of absorptive surface rather than on the position of the resected segment. The bypass loop may itself still exert some influence on lipoprotein metabolism, mainly on high density lipoprotein-cholesterol.     

Modifications of enzyme activities in rat liver plasma membrane after carbon tetrachloride poisoning

Adenylate Cyclase activity is increased in the liver of animals treated with CCl4 (250 ul/100g body wt.) after 30 min. The maximum increase is observed 2 hours after administration of the hepatotoxin. Whereas, 3',5'-nucleotidephosphodiesterase decreases significantly throughout all the experiments. Our results present evidence that there is relationship between Adenylate Cyclase and Phosphodiesterase activity and suggest that intracellular calcium ion may mediate a regulation of the synthesis and degradation of cyclic nucleotides. It is difficult to determine the exact role Ca2+ plays in regulating these two opposing reactions. Thus, in the near future the work in this laboratory will be to define carefully the effects of the CCl4 on ions on these two systems.     

Adipogenic activities in rat organs

Adipose conversion of 3T3-L1 cells depends on adipogenic factors present in serum. In order to find their origin, adipogenic activity in extracts from adrenals, kidneys, testes, ovaries, liver, spleen, sceletal muscle and adipose tissue of the rat was studied. If tissues were homogenized with aqueous buffers at low pH, no adipogenic activity could be extracted. Treatment with chloroform/methanol followed by phase separation however revealed considerable adipogenic activity in the organic phase from adrenals, kidneys, ovaries, testes and sceletal muscle and additionally in the aqueous phase of liver. Further purification by liquid chromatography and reversed phase HPLC led to the identification of adipogenic activities from adrenals and kidneys as corticosterone and 11-dehydrocorticosterone. Adipogenic activity from liver in contrast is pronase-sensitive, exhibits an apparent molecular mass of 4 kDa and is probably a peptide.     

Lesion bypass activities of human DNA polymerase mu

DNA polymerase mu (Polmu) is a newly discovered member of the polymerase X family with unknown cellular function. The understanding of Polmu function should be facilitated by an understanding of its biochemical activities. By using purified human Polmu for biochemical analyses, we discovered the lesion bypass activities of this polymerase in response to several types of DNA damage. When it encountered a template 8-oxoguanine, abasic site, or 1,N(6)-ethenoadenine, purified human Polmu efficiently bypassed the lesion. Even bulky DNA adducts such as N-2-acetylaminofluorene-adducted guanine, (+)- and (-)-trans-anti-benzo[a]pyrene-N(2)-dG were unable to block the polymerase activity of human Polmu. Bypass of these simple base damage and bulky adducts was predominantly achieved by human Polmu through a deletion mechanism. The Polmu specificity of nucleotide incorporation indicates that the deletion resulted from primer realignment before translesion synthesis. Purified human Polmu also effectively bypassed a template cis-syn TT dimer. However, this bypass was achieved in a mainly error-free manner with AA incorporation opposite the TT dimer. These results provide new insights into the biochemistry of human Polmu and show that efficient translesion synthesis activity is not strictly confined to the Y family polymerases.