Occurrence of polyamine oxidase in tissues of the catfish

1. Polyamine oxidase (PAO) activity was found in the brain, intestine, kidney and liver of the siluroid catfish using N1-acetylspermine as substrate. It was highest in the intestine and lowest in the brain. 2. Substrate specificity of the enzyme was tested in the intestine and liver and the highest activity was found with N1-acetylspermine, followed by N1-acetylspermidine and N1,N12-diacetylspermine. 3. The apparent Km values for N1-acetylspermine were 19.6 and 46.9 microM for the intestine and liver, respectively. 4. These results suggest the presence of a system of polyamine reutilization after their acetylation in fishes.     

Effect of dietary fiber on microbial activity and microbial gas production in various regions of the gastrointestinal tract of pigs.

The microbial activity, composition of the gas phase, and gas production rates in the gastrointestinal tract of pigs fed either a low- or a high-fiber diet were investigated. Dense populations of culturable anaerobic bacteria, high ATP concentrations, and high adenylate energy charges were found for the last third of the small intestine, indicating that substantial microbial activity takes place in that portion of the gut. The highest microbial activity (highest bacterium counts, highest ATP concentration, high adenylate energy charge, and low pH) was found in the cecum and proximal colon. Greater microbial activity was found in the stomach and all segments of the hindgut in the pigs fed the high-fiber diet than in the pigs fed the low-fiber diet. Considerable amounts of O2 were found in the stomach (around 5%), while the content of O2 in gas samples taken from all other parts of the gastrointestinal tract was < 1%. The highest concentrations and highest production rates for H2 were found in the last third of the small intestine. No methane could be detected in the stomach or the small intestine. The rate of production and concentration of methane in the cecum and the proximal colon were low, followed by a steady increase in the successive segments of the hindgut. A very good correlation between in vivo and in vitro measurements of methane production was found. The amount of CH4 produced by pigs fed the low-fiber diet was 1.4 liters/day per animal. Substantially larger amounts of CH4 were produced by pigs fed the high-fiber diet (12.5 liters/day)(ABSTRACT TRUNCATED AT 250 WORDS)     

春季树麻雀体内几种消化酶活性研究

应用酶学分析法测定了树麻雀Passer montanus春季的腺胃、肌胃、小肠、大肠、肝脏和胰脏组织中淀粉酶、蛋白酶和纤维素酶的活力.结果表明,不同组织中的消化酶活力差异显著,淀粉酶和蛋白酶以胰脏中活力最高,腺胃次之,纤维素酶均较低;同一组织中不同消化酶的活力差异显著,淀粉酶活性最高,蛋白酶次之,纤维素酶活力最低.这些差异提示消化酶活力大小与器官分化有关,并受食物组成的影响,因此产生了不同的酶活力分布.这是树麻雀长期适应东北地区寒冷环境的生存策略之一.     

The cellular distribution of the metabolic deamination of benzylamine

The metabolism of benzylamine was investigated using the 600g supernatant, mitochondrial, microsomal and cytosol fractions of different rat organs and the livers of various animal species. This substrate was extensively deaminated to benzaldehyde, benzyl alcohol and benzoic acid. The ratio of the metabolic products formed varied greatly depending on the nature of the homogenate used in the incubation mixture of benzylamine. The specific activity of the deamination reaction was mainly concentrated in the mitochondrial and microsomal fractions. In many organs, the microsomal preparations were more active than the mitochondria. The liver was the rat organ with the highest deaminating activity. Hepatic homogenates from rabbit were the most active amongst similar fractions from other animal species. The N-oxygenated products, N-hydroxybenzylamine and benzaldoxime, could not be isolated from the incubation mixtures of benzylamine.     

The response of digestive proteases to abrupt salinity decrease in the euryhaline sparid Sparus aurata L

The response of the digestive proteases to abrupt salinity change was studied in juvenile gilthead sea bream (Sparus aurata) for 15 days after transfer from 33 per thousand to 21 per thousand. Salinity decrease affected significantly neither the activity of total acid proteases in stomach, nor the activities of total alkaline proteases and major serine proteases--trypsin and chymotrypsin--in the alkaline part of the intestine. The activity of the major proteases was significantly different between the alkaline segments of the intestine, with the posterior intestine presenting the highest activities followed by the pyloric caeca. This distribution pattern remained unaffected by salinity decrease. Notably, salinity change led to significant alterations in elastase and carboxypeptidase activity. The changes were more prominent in the upper part of the intestine (pyloric caeca and anterior intestine) than in the posterior intestine. In pyloric caeca significant alteration of carboxypeptidase A and B activities was observed, elastase changes were confined to anterior intestine together with alterations in carboxypeptidase B activity, while in posterior intestine the changes were restricted to carboxypeptidase A activity. The results are discussed in relation to the osmoregulatory action of the intestinal segments and dietary protein digestion.     

Carbohydrases in the digestive tract of the African bony-tongue Heterotis niloticus (Pisces: Osteoglossidae)

The presence of amylase and maltase in the stomach, intestine and pyloric caeca of Heterotis niloticus is demonstrated. Amylase activity was highest in the fore-gut, followed by the pyloric caeca, while the lowest activity was in the hind-gut. Optimum pH for intestinal amylase was 8.45. Sucrase, lactase and cellulase were not detected.     

The appearance of 1,25-dihydroxyvitamin D3 receptor during chick embryo development

The appearance of the 1,25-dihydroxyvitamin D3 receptor in intestine, kidney, and chorioallantoic membrane of chick embryo was followed by sucrose density gradient sedimentation analysis and Scatchard plot analysis. The receptor from each of these organs sediments as a single 3.7S component. At 19 days of embryonic life, intestine had the highest specific 1,25-dihydroxyvitamin D3 binding activity followed by kidney and chorioallantoic membrane. The 1,25-dihydroxyvitamin D3 binding activity increased gradually at 12-15 days and rapidly until 20 days in intestine. In kidney, this protein increased rapidly from 12 to 16 days and did not change subsequently. In chorioallantoic membrane, the receptor increased slowly from 8 through 15 days, rapidly until 19 days, and decreased at 20 days. The injection of hydrocortisone into the chick embryo at 10 days increased receptor number in intestine, kidney, and chorioallantoic membrane by a factor of 2 at 12 days. Injection of this hormone after this time had little or no effect.     

Influence of age on Cu/Zn-superoxide dismutase and indole 2,3-dioxygenase activities in rat tissues

The aim of this study was to investigate in vitro the variations with age of the activities of the two antioxidant enzymes Cu/Zn-superoxide dismutase (SOD) and indole 2,3-dioxygenase (IDO) in metabolically active tissues of rats of various ages. In rats aged one week and 2-3 months the highest Cu/Zn-SOD activity was found in the liver and the lowest in the small intestine. At 12 and 18 months of age, the activity was higher in the brain and kidneys, when compared to the small intestine, lungs and liver. Cu/Zn-SOD activity decreased significantly after 2-3 months of age with advancing age in all tissues examined. In newborn rats IDO activity was present only in the small intestine. In the group of rats aged 2-3 months, the highest specific activity was observed in the small intestine and the lowest in the lungs and kidneys, whereas at 12 months of age, the highest IDO activity was found in the brain, with kidneys presenting the lowest activity. At 18 months, IDO returned to be more elevated in the small intestine. At 12 months of age the values of IDO in the tissues varied slightly, while at 18 months similar activities were found between the lungs and brain and between the small intestine and kidneys. In relation to age, IDO specific activity declined in the small intestine, after 2-3 months of age. In the lungs, the activity remained unchanged; in the brain and in the kidneys activity decreased significantly from 2-3 to 18 months of age. In conclusion, this study demonstrates an age-related decline in Cu/Zn-SOD and IDO activities, the two enzymes responsible for scavenging O2*-.     

Novel copper amine oxidase activity from rat liver mitochondria matrix

Copper containing amine oxidases (Cu-AO) represent a heterogeneous class of enzymes classified as EC 1.4.3.6. The present study reports preliminary results on the presence of a novel amine oxidase activity in rat liver mitochondria lysates. Such enzymatic activity was found in the soluble mitochondrial fraction, obtained by simple osmotic shock. The mitochondrial amine oxidase was isolated by affinity chromatography on a newly synthesised spermine-Sepharose. SDS-PAGE showed a single band at about 60 kDa. Upon chromatographic purification, the enzymatic activity was very labile. The crude enzyme activity was tested by spectrophotometric measurements, determining hydrogen peroxide production following oxidative deamination of different substrates, such as polyamines (spermine, spermidine, putrescine and cadaverine) and monoamines (dopamine and benzylamine). The activity, observed on polyamines and not on monoamines, was inhibited by semicarbazide and azide, but not by pargyline, clorgyline and l-deprenil. Enzyme specificity was tested on several diamines characterized by different carbon atom chain length in the range 2-6 carbon atoms. The highest activity was found with 1,2-diamino-ethane and the highest affinity with 1,5-diamino-pentane. The above reported results suggest the presence of a novel copper-dependent amine oxidase in liver mitochondria matrix.     

Hydrogenosomes in known species of rumen entodiniomorphid protozoa

Abstract Yeasts of the genus Kluyveromyces grew very slowly on methylamine as sole nitrogen source. Methylamine oxidase activity in cell-free extracts was very low. Under conditions known to separate methylamine oxidase from benzylamine oxidase in other yeast genera, only a single enzyme was detected in Kluyveromyces lactis . This enzyme could oxidize benzylamine, n -butylamine and (very poorly) methylamine. The enzyme lost no activity on heating at 45°C and had a high affinity and V _max for benzylamine and 1-aminoalkanes of long-chain length, with a very low affinity and V _max for methylamine. It is concluded that growth of K. lactis on methylamine involves only benzylamine oxidase, and that a methylamine oxidase of the type found in other yeasts does not occur.     

Characterization of heme oxygenase in the small intestinal epithelium

Heme oxygenase activity was examined in the epithelial cells of the small intestine in male Sprague-Dawley rats. As with liver and spleen, the highest specific activity of this enzyme was found in the microsomal fraction of these cells. Substrate kinetics, analysis of cofactor requirements, and other biochemical characteristics suggested further similarities between heme oxygenase in the small intestine and liver. Enzyme activity was differentially localized longitudinally within the small intestine, with the highest specific activity occurring in the region approximately 15 to 30 cm beyond the pylorus. The effects of diet on the basal levels of heme oxygenase in the proximal small intestine were also examined. Although intestinal cytochrome P450-dependent monooxygenase activity, as determined by benzo[a]pyrene hydroxylase and 7-ethoxycoumarin O-deethylase, was greatly reduced (65-90%) in animals maintained on a semipurified control diet compared with standard cereal-based chow, there were no differences observed in heme oxygenase activity between the two dietary treatment groups. The activity of intestinal heme oxygenase could be increased, however, by oral treatment with several metal compounds that are known to affect hepatic heme metabolism when administered parenterally. The enzyme activity was also potently inhibited by tin (Sn4+) protoporphyrin administered orally or parenterally.     

Control of benzylamine oxidase activity in Kluyveromyces fragilis grown on primary amines as nitrogen source

Abstract After growth on a mixture of ammonium and either methylamine or n -butylamine as nitrogen sources, benzylamine oxidase activity in yeasts from a number of different genera was found to be repressed to a lesser extent by ammonium than was methylamine oxidase. Catalase activity was better repressed by ammonium with methylamine as the nitrogen source than with n -butylamine. During growth of Kluyveromyces fragilis on equimolar mixtures of ammonium and an amine as nitrogen sources, benzylamine oxidase synthesis began during the period of exclusive growth on ammonium, and a period of simultaneous use of both nitrogen sources was observed just before the ammonium was exhausted. Addition of ammonium to cultures growing on n -butylamine as nitrogen source had no immediate repressive effect on benzylamine oxidase or catalase synthesis. However, growth on limiting ammonium in the absence of amines did give rise to low levels of amine oxidase and derepression of catalase activity. It is concluded that benzylamine oxidase in yeasts is induced strongly by amines as well as being less strongly repressed by ammonium than methylamine oxidase.     

Antiproliferative and anticytotoxic effects of cell fractions and exopolysaccharides from Lactobacillus casei 01

Cell fractions including heat-treated cells, crude cell walls, intracellular extracts and exopolysaccharides (EPSs) obtained from Lactobacillus casei 01 were first studied for their effects on the proliferation of human intestinal epithelial cells, intestine 407 and the human colon cancer cell, HT-29. Their effects on the cytotoxicity of 4-nitroquinoline 1-oxide (4-NQO) against intestine 407 were further investigated. The results revealed that EPS exhibited the highest antiproliferation activity on HT-29 cells while the viability of intestine 407 cells was not affected by EPS at a concentration of 5-50μg/mL. It was also noted that all the cell fractions and EPS from L. casei 01 reduced the cytotoxicity of 4-NQO against intestine 407 with EPS showing the highest anticytotoxic activity. Additionally, it was found that EPS might exert blocking and bioanticytotoxic effects by both adjusting the function of intestine 407 and repairing the 4-NQO-damaged cells, thus reducing cytotoxicity of 4-NQO.     

Studies on the formation of gamma-aminobutyric acid from putrescine in rat organs and purification of its synthetic enzyme from rat intestine.

The formation of gamma-aminobutyric acid (GABA) from putrescine was examined in organs of rats using radioactive putrescine. Radioactive GABA was detected in all the organs of a rat injected intraperitoneally with radioactive putrescine, and the highest radioactivity of GABA was observed in the small intestine. The enzyme involved in this formation was purified from small intestine and identified as a diamine oxidase, histaminase, from the properties of the enzyme. Activity of the enzyme was found in all the organs of rat, and the highest activity was observed in the small intestine.     

Histochemical localization of glucose-6-phosphatase and 5-nucleotidase in the digestive system of Ophiocephalus Channa punctatus.

The histochemical localization of G6Pase and 5-Nase in the digestive system of Ophiocephalus (Channa) punctatus was studied. The highest activities of these enzymes were found in the liver. Appreciable activity was also found in the anterior intestine (duodenum) and pyloric caeca. The activity faded toward the middle and posterior intestine and rectum. In the stomach the activity was moderate. The activity of 5-Nase was weaker than that of G6Pase. In the stomach the enzymes were localized in the mucosa and gastric glands. The absorptive columnar epithelial cells were the major sites of localization in the intestine. The goblet cells were negative. The G6Pase activity was associated with the cytoplasm, while the 5-Nase activity was found in the cell membranes and the nuclei.     

Glutamine synthetase activity in the organs of fed and 24-hours fasted rats

Glutamine synthetase activity in several rat tissues had been measured. Liver contains the highest specific activity followed by stomach, brain, kidneys, intestine, skin, adipose tissue and striated muscle - that had the lowest specific activity both with regard to tissue, protein and DNA weight. Per unit of animal weight, liver and muscle contain similar activities, 24 Hours of fasting induced a significant decrease in liver, stomach, intestine and skin glutamine synthetase, compensated by an increase in muscle activity. During fasting, the splanchnic glutamine synthetase activity is lowered and that of peripheral tissues is increased, thus favoring a net glutamine flux from peripheral to splanchnic organs.     

Dietary regulation of adenosine deaminase activity in stomach, small intestine and spleen of mice

Activity of adenosine deaminase (ADA) and its regulation by dietary restriction were studied in the stomach, small intestine and spleen of mice. ADA activity (U/mg protein) was highest in the stomach, followed by small intestine and spleen of mice on normal diet. The activity decreased significantly in the stomach (41%) and small intestine (45%) of 24 hr fasted mice, when compared to mice fed ad-libitum. However, ADA activity in spleen did not show any change by dietary intervention. Refeeding of fasted mice for 24 hr restored the activity of ADA in tissues. In addition, dietary restriction (alternate days of feeding for three months) had a cumulative effect, whereby ADA activity decreased significantly in the stomach (53% on the day of feeding and 60% on the day of fasting) and small intestine (50% and 54% on the day of feeding and fasting, respectively) without any change in activity in spleen. These findings indicate that dietary restriction reduces ADA activity in a tissue-specific manner. Long-term dietary restriction leads to a cumulative adaptation in lowering the ADA activity of GIT, but not in spleen.     

Antifeedant activity and toxicity of two alkaloids from Adhatoda vasica Nees leaves against diamondback moth Plutella xylostella (Linn.) (Lepidoptera: Plutellidae) larvae

Two natural alkaloids viz., Vasicine acetate and 2-Acetyl benzylamine, isolated from Adhatoda vasica leaves, showed antifeedant, larvicidal and moult inhibiting properties against diamondback moth Plutella xylostella in laboratory experiments. Maximum antifeedant activity of 98.5% was recorded at 1000 ppm concentration of Vasicine acetate treatment, whereas as 2-Acetyl benzyl amine recorded only 71.4% antifeedant activity at 1000 ppm concentration. Azadirachtin treatment presented 82% antifeedant activity at the highest concentration (1000 ppm). Both the active compounds of A. vasica showed lethal toxicity on larvae and pupae. The highest larvicidal and pupicidal activities were recorded in 2-Acetyl benzylamine treatment at 125 ppm concentration. The two A. vasica compounds also affected the normal growth and development and moulting process of P. xylostella. Final moulting of larvae into pupae was disrupted by the treatments, which resulted in larval–pupal intermediates and abnormal pupae. Treatments also produced small-size pupae and malformed adults with poorly developed wings.     

Intestinal arginase in vertebrates and invertebrates.

1. Arginase was found to be present in the intestine in all species of Annelida, Arthropoda and Chordata studied. 2. The activity of intestinal arginase differs from species to species, the differences reaching two orders of magnitude (100 x). 3. The highest activity of intestinal arginase was observed in the rodents (mouse, rat, hamster). 4. In animals in which the enzyme activity was high or moderately high, arginase activity showed topographical differentiation along the long axis of the intestine.     

白条草蜥消化道内分泌细胞的免疫组织化学

应用6种胃肠激素抗血清和免疫组织化学ABC法(avidin-biotin complex method),对白条草蜥(Takydromus wolteri)消化道内分泌细胞进行了免疫组织化学定位研究和形态学观察。结果表明,5-羟色胺细胞较其他5种内分泌细胞的分布更为广泛,整个消化道中(即从食管到直肠)均有分布,其分布密度高峰位于幽门。食管、回肠和直肠未检测到生长抑素细胞,生长抑素细胞在幽门部分布密度最高,总体来说生长抑素细胞的分布在胃部较高而在小肠部较低。胃泌素细胞和胰多肽细胞分布在小肠,均在十二指肠分布密度最高。胰高血糖素细胞在胃幽门部分布密度最高,十二指肠、空肠次之,回肠分布密度最低。P-物质细胞仅分布于幽门部。6种内分泌细胞以圆形和锥体形为主,它们广泛分布于消化道黏膜之间、腺泡上皮细胞之间及上皮细胞基部。内分泌细胞的密度分布与其食性、食物组成和生活环境有关,它们的形态与其内、外分泌功能是相适应的。