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The effects of varied durations of food deprivation on the rates and kinetics of glucose phosphorylation by isolated rat hepatocytes have been examined. Glucokinase activity was measured concurrently in extracts from these cells prepared from livers of rats which had fasted for 0, 24, 48 and 72 h. Significant levels of hepatocyte glucose phosphorylation were noted even when glucokinase levels were extrapolated to zero. The K0.5-glucose value of 33 mM in cells from fed rats increased to 48 mM after a 72-h fast. It is concluded that a high K0.5 glucose-phosphorylating enzyme or enzymes compensatory to insulin-dependent glucokinase is/are involved in rat liver glucose phosphorylation.  相似文献   

3.
As with other regeneration processes, adventitious root formationmay be divided into three phases, namely dedifferentiation,induction and differentiation. We assumed that the appropriatehormonal conditions for rooting, in particular a high levelof auxin and a low level of cytokinin, are required only duringthe induction phase. Hence, the effect of 24 h pulses with indolebutyricacid (IBA) or benzylaminopurine (BAP) should be maximal duringthis phase. On this assumption, the timing of the three phaseswas determined in microcuttings of Malus ‘Jork 9’.The promotion of rooting was largest for 24 h IBA-pulses given24–48 h or 48–72 h after the onset of culture. Duringculture of shoots on IBA-containing medium 24 h BAP-pulses weregiven. Inhibition of rooting was maximal for the BAP-pulsesgiven between 24 and 96 h. An analysis of the kinetics of rootemergence also indicated that added IBA acted from 24 h onwards:emergence of roots from the tissue occurred simultaneously forthe IBA pulses at 0–24 h and 24–48 h, but lagged1 d behind for the 48–72 h pulse and 2 d for the 72–96h pulse. We concluded that dedifferentiation occurred from 0to 24 h, induction from 24 to 72 or 96 h, and after that differentiation.Histological observations showed that after 24 h, cells withswollen nuclei and dense cytoplasm had appeared in the regionsof the stem where the roots were formed. The first cell divisionswere observed after 48 h. After 96 h, meristemoids of c. 30cells had been formed. After the BAP-pulses at 24–48 hor 48–72 h, these meristematic cells formed callus. Key words: Adventitious rooting, apple, auxin, cytokinin, Malus, regeneration.  相似文献   

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为探究取食施硅水稻后白背飞虱Sogatella furcifera体内解毒酶(羧酸酯酶(CarE)、谷胱甘肽-S-转移酶(GST)、多功能氧化酶(MFO))和保护酶(超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD))活性的变化。以感虫品种TN1为供试水稻品种,采用营养液添加硅酸钠(Na2SiO3·9H2O)的方法设置施硅(Si+, 112 mg Si/L)和不施硅(Si-, 0 mg Si/L)处理,测定取食Si+或Si-水稻24 h、48 h、72 h和96 h后白背飞虱成虫体内CarE、GST、MFO、SOD、CAT和POD活性的动态变化。结果显示,与取食Si-水稻的白背飞虱相比,取食Si+水稻48 h、72 h和96 h时,CarE活性显著降低,GST活性显著升高;取食72 h和96 h时,MFO活性显著上升。取食Si+水稻24 h和48 h时,SOD和CAT活性显著增大;取食72 h时,SOD和POD活性显著增高;取食96 h时,3种保护酶活性均降低,其中SOD和POD活性显著降低。以上结果表明,施硅可能通过影响白背飞虱体内解毒酶和保护酶活性的动态平衡,进而对白背飞虱生理代谢造成影响。本研究为合理利用硅肥防治白背飞虱提供了理论依据。  相似文献   

6.
1. The effects of dietary modification, including starvation, and of corticotropin injection on the activities of acyl-CoA synthetase, glycerol phosphate acyltransferase, dihydroxyacetone phosphate acyltransferase, phosphatidate phosphohydrolase, diacylglycerol acyltransferase and lipoprotein lipase were measured in adipose tissue. 2. Lipoprotein lipase activities in heart were increased and those in adipose tissue were decreased when rats were fed on diets enriched with corn oil or beef tallow rather than with sucrose or starch. The lipoprotein lipase activity was lower in the adipose tissue of rats fed on the sucrose rather than on the starch diet. 3. Rats fed on the beef tallow diet had slightly higher activities of the total glycerol phosphate acyltransferase in adipose tissue than did rats fed on the sucrose or starch diet. The diacylglycerol acyltransferase and the mitochondrial glycerol phosphate acyltransferase activities were higher for the rats fed on the tallow diet than for those fed on the corn-oil diet. 4. Starvation significantly decreased the activities of lipoprotein lipase (after 24 and 48 h), acyl-CoA synthetase (after 24 h) and of the mitochondrial glycerol phosphate acyltransferase and the N-ethylmaleimide-insensitive dihydroxyacetone phosphate acyltransferase (after 48 h) in adipose tissue. The activities of the microsomal glycerol phosphate acyltransferase, diacylglycerol acyltransferase and the soluble phosphatidate phosphohydrolase were not significantly changed after 24 or 48 h of starvation. 5. The activities of lipoprotein lipase and phosphatidate phosphohydrolase in adipose tissue were decreased 15 min after corticotropin was injected into rats during November to December. No statistically significant differences were found when these experiments were performed during March to September. These differences may be related to the seasonal variation in acute lipolytic responses. 6. These results are discussed in relation to the control of triacylglycerol synthesis and lipoprotein metabolism.  相似文献   

7.
Synthetic rates of fatty acid, cholesterol and triacylglycerols, and contents and secretion of lipoprotein lipids, were determined in hepatocytes of rats fed ad libitum a fat-containing stock diet or of rats fasted for 48 h and then refed for 24 or 48 h with stock diet or with a glucose-rich fat-free diet. When compared with the values for the ad libitum-fed rats, fatty acid synthesis was lower in fasted rats, slightly increased in rats refed with the stock diet, but several-fold elevated after refeeding the glucose-rich fat-free diet. Cholesterol synthesis was decreased in the fasted cells, and restored to the control level upon refeeding either diet. Triacylglycerol synthesis from exogenous oleate was greatly stimulated in the cells of fasted-refed rats above the rate in cells of the ad libitum-fed rats, the increase being considerably higher after refeeding the glucose-rich fat-free diet than the stock diet. The amount of triacylglycerol secreted by the cells was also elevated by the fasting-refeeding treatment, but the difference between the two diets was much less pronounced than seen for the lipids' synthetic rates. This imbalance may underlie the huge accumulation of this lipid observed in the heptatocytes after refeeding the rats for 48 h with the glucose-rich fat-free diet.  相似文献   

8.
Heat-inactivated serum is cytotoxic to granulosa cells from preantral follicles but not to cells from preovulatory follicles. A dominant feature of the granulosa cells of preovulatory follicles is the presence of luteinizing hormone (LH) receptors on the surface of the cells. In the present study, we have examined the relationship between the process of LH receptor induction and the acquisition of serum tolerance in granulosa cells in vitro. Granulosa cells from the ovaries of immature rats primed with diethylstilbestrol (DES) were cultured in a 1:1 mixture of Ham's F-12 and Dulbecco's modified Eagle's medium containing 30 ng of ovine follicle-stimulating hormone (oFSH; NIH-15). At either 0, 24, or 48 h of culture, heat-inactivated fetal bovine serum (FBS) was added (10% by volume) to separate groups of culture tubes. All cells were cultured for a total of 72 h, at which time the cultures were assessed for LH receptor (specific 125I-human chorionic gonadotropin [hCG] binding) and DNA content. LH receptors were induced in all FSH-containing serum-free cultures by 48 h. Receptors were not induced, however, when serum was added after either 0 or 24 h of culture. Furthermore, serum addition at these times resulted in a cell loss (assessed by DNA) of 40-60%. Serum addition at 48 h to FSH-containing cultures resulted in an inability to detect LH receptors at 72 h and with no significant effect on the culture DNA content. Addition of a protein extract of FBS at the initiation of cell culture prevented FSH-stimulated LH receptor induction and was cytotoxic. A lipid extract of FSH did not interfere with receptor induction and was not cytotoxic.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

9.
A morphological study of the midgut of Lutzomyia intermedia, the primary vector of cutaneous leishmaniasis, in southeast Brazil, was conducted by light, scanning and transmission electron microscopy. The midgut is formed by a layer of epithelium of columnar cells on a non-cellular basal lamina, under which there is a musculature, which consists of circular and longitudinal muscular fibers. A tracheolar network is observed surrounding and penetrating in the musculature. Females were examined 12, 24, 48, 72 h and 5 days following a blood meal and were analyzed comparatively by transmission electron microscopy with starved females. In starved females, the epithelium of both the anterior and posterior sections of the midgut present whorl shaped rough endoplasmic reticulum. The posterior section does not present well-developed cellular structures such as mitochondria. Observations performed at 12, 24, 48 and 72 h after the blood meal showed morphological changes in the cellular structures in this section, and the presence of the peritrophic matrix up to 48 h after the blood meal. Digestion is almost complete and a few residues are detected in the lumen 72 h after blood feeding. Finally, on the 5th day after the blood meal all cellular structures present the original feature resembling that seen in starved sand flies. Morphometric data confirmed the morphological observations. Mitochondria, nuclei and microvilli of midgut epithelial cells are different in starved and blood fed females. The mitochondria present a similar profile in the epithelium of both the anterior and posterior section of the midgut, with higher dimension in starved females. The cell microvilli in the posterior section of the midgut of starved females are twice the size of those that had taken a blood meal. We concluded that there are changes in the midgut cellular structures of L. intermedia during the digestion of blood, which are in agreement with those described for other hematophagous diptera.  相似文献   

10.
The effects of varied durations of food deprivation on the rates and kinetics of glucose phosphorylation by isolated rat hepatocytes have been examined. Glucokinase activity was measured concurrently in extracts from these cells prepared from livers of rats which had fasted for 0, 24, 48 and 72 h. Significant levels of hepatocyte glucose phosphorylation were noted even when glucokinase levels were extrapolated to zero. The K0.5-glucose value of 33 mM in cells from fed rats increased to 48 mM after a 72-h fast. It is concluded that a high K0.5 glucose-phosphorylating enzyme or enzymes compensatory to insulin-dependent glucokinase is/are involved in rat liver glucose phosphorylation.  相似文献   

11.
Female mosquitoes feed on human blood, which can be collected to analyze human short tandem repeat (STR) sequences; these are specific to each human individual. Analysis of STRs might help in identification of a person found near a crime scene. Aedes aegypti and Culex pipiens mosquitoes fed on human blood were cultured at 18°C or 40°C (median temperature for summer and winter time in Riyadh governorate, Saudi Arabia) for 3, 6, 12, 24, 48 and 72 h. In A. aegypti, human DNA concentration was reduced with time at both temperatures. At 18°C, we obtained full STR profiles up to 48 h post feeding on human blood while none of the 16 loci were obtained at 72 h. At 40°C, we missed six sites at 12 h after blood sucking, 12 at 24 h, and 15 at 48 h and 72 h. In C. pipiens cultured at 18°C, full profiles were developed up to 48 h following blood feeding while we could not amplify five sites at 72 h. At 40°C, mortality among females was 50% at 24 h and 100% at both 48 h and 72 h; however, we had full profiles in all samples including dead insects. This research addressed the possibility of using mosquitoes in forensic research by DNA genotyping by changing the mosquito culturing temperature and mosquito genus. Our findings proved that different types of mosquito change the temporal pattern of STR analysis and showed that the mosquito culturing temperature affects the integrity of DNA for STR analysis.  相似文献   

12.
Betanodavirus protein alpha induces cell apoptosis or secondary necrosis by a poorly understood process. In the present work, red spotted grouper nervous necrosis virus (RGNNV) RNA 2 was cloned and transfected into tissue culture cells (GF-1) which then underwent apoptosis or post-apoptotic necrosis. In the early apoptotic stage, progressive phosphatidylserine externalization was evident at 24h post-transfection (p.t.) by Annexin V-FLUOS staining. TUNEL assay revealed apoptotic cells at 24-72 h p.t, after which post-apoptotic necrotic cells were identified by acridine orange/ethidium bromide dual dye staining from 48 to 72 h p.t. Protein alpha induced progressive loss of mitochondrial membrane potential (MMP) which was detected in RNA2-transfected GF-1 cells at 24, 48, and 72 h p.t., which correlated with cytochrome c release, especially at 72 h p.t. To assess the effect of zfBcl-xL on cell death, RNA2-transfected cells were co-transfected with zfBcl-x(L). Co-transfection of GF-1 cells prevented loss of MMP at 24 h and 48 h p.t. and blocked initiator caspase-8 and effector caspase-3 activation at 48 h p.t. We conclude that RGNNV protein alpha induces apoptosis followed by secondary necrotic cell death through a mitochondria-mediated death pathway and activation of caspases-8 and -3.  相似文献   

13.
The immune system has been recognized as a potential contributor to psychiatric disorders. In animals, lipopolysaccharide (LPS) is used to induce inflammation and behaviors analogous to some of the symptoms in these disorders. Recent data indicate that the kynurenine pathway contributes to LPS-induced aberrant behaviors. However, data are inconclusive regarding optimal LPS dose and treatment strategy. Here, we therefore aimed to evaluate the effects of single versus repeated administration of LPS on the kynurenine pathway. Adult C57BL6 mice were given 0.83 mg/kg LPS as a single or a repeated injection (LPS + LPS) and sacrificed after 24, 48, 72, or 120 h. Mice receiving LPS + LPS had significantly elevated brain kynurenine levels at 24 and 48 h, and elevated serum kynurenine at 24, 48 and 72 h. Brain kynurenic acid and quinolinic acid were significantly increased at 24 and 48 h in mice receiving LPS + LPS, whereas serum kynurenic acid levels were significantly decreased at 24 h. The increase of brain kynurenic acid by LPS + LPS was likely unrelated to the higher total dose as a separate group of mice receiving 1.66 mg/kg LPS as single injection 24 h prior to sacrifice did not show increased brain kynurenic acid. Serum quinolinic acid levels were not affected by LPS + LPS compared to vehicle. Animals given repeated injections of LPS showed a more robust induction of the kynurenine pathway in contrast to animals receiving a single injection. These results may be valuable in light of data showing the importance of the kynurenine pathway in psychiatric disorders.  相似文献   

14.
Zinc deficiency leads to decreased cellular immune responses. The overproduction of nitrogen species derived from inducible nitric oxide synthase (iNOS), its enzyme, and interleukine-1 beta (IL-1β), and inflammatory cytokine have been implicated in immune responses. The goal of this study was to investigate the effects of lipopolysaccharide (LPS)-induced changes in NO metabolites, iNOS, and IL-1β protein expression in the lungs of zinc-deficient rats. Male Sprague–Dawley rats (body weight, 100 g) were divided into two groups and were fed either a zinc-deficient diet (ZnD) or a zinc-containing diet (Cont). After 4 weeks on these diets, rats received a 10-mg/kg dose of LPS injected via the tail vein and were then maintained for an additional 72 h. To determine total NO concentrations in the blood, serum zinc concentration, iNOS protein expression, IL-1β, and iNOS immunohistochemistry, blood and lung samples were obtained at pre-LPS injection, 5, 24, and 72 h after injection. Total NO levels were significantly increased at 5, at 24, and at 72 h after LPS injection compared with pre-LPS injection level in ZnD group; significant changes in total NO levels was elevated at 5 h from at pre-LPS level but not significant changes from basal level at 24 and 72 h in the control group. Based on western blot analyses and immunohistochemistry, clear bands indicating iNOS and IL-1β protein expression and iNOS antibody-stained inflammatory cells were detected at 5 and 24 h in the ZnD group and 5 h in the Cont group, not observed at 24 and 72 h in the control group. These results suggest that zinc deficiency induces overexpression of iNOS and IL-1β proteins from inflammatory cells around the alveolar blood vessels, resulting in overproduction of total NO and persisted inflammatory response in the zinc-deficient rat lung. Taken together, overexpression of LPS-induced iNOS, overproduction of iNOS-derived NO, and overexpression of IL-1β may induce nitrosative and oxidative stresses in the lung, and these stresses may be involved low immunity of zinc deficiency states.  相似文献   

15.
The effect of neutron doses from a D-T compact neutron generator on the liver cells of adult male and female albino Swiss mice was investigated. Fast neutrons (14.5 MeV) were delivered to the whole body in a single dose or in two, four, six or eight equal doses separated by 3-day intervals. The lowest dose, 100 rem, was given for an exposure time of 6 hours and was then steadily raised to 912 rem for an exposure time of 48 hours. During exposure the neutron flux was controlled by the activation foil technique. Animals were killed for testing after each irradiation. Histological examination of the hepatocytes in the light microscope showed marked degenerative changes only after the longer irradiation periods (24, 36 and 48 h). Electron microscopy showed cytological (cytoplasmic and nuclear) changes in the hepatocytes after only 12 hours' irradiation. Densitometric scans of electron micrographs of control and 12 h-irradiated livers indicated that the control hepatocyte interphase nucleus contains approximately 72% heterochromatin, while the irradiated nucleus contains only 64% heterochromatin.  相似文献   

16.
After 16 h nocturnal deprivation of food, male Wistar rats were irradiated by a single whole body dose of 2.40 Gy X-rays. Both the irradiated and sham-irradiated (control) rats were pair-fed for the first six days after irradiation, but for the rest of the time they were fed ad libitum. Lipoprotein lipase activity (LPLA) in the adipose tissue fell between 24 and 48 h; LPLA in the heart fell at 24 h and 21 days and rose on the 14th days. The serum triacylglycerol concentration rose between 24 and 72 h. Comparison with the fed control group showed LPLA in adipose tissue to be reduced at 6 and 72 h and on the 28th day and raised between the 7th and the 14th day. In the heart it was raised at 1 h and between 72 h and the 14th day, it was reduced on the 21st day and rose on the 35th day. The triacylglycerol concentration was raised between 48 and 72 h and on the 28th day. Pair-feeding after non-lethal X-irradiation allowed more exact differentiation of the specific effect of ionizing radiation on LPLA in the adipose tissue and heart at the early post-irradiation intervals.  相似文献   

17.
The growth of L fibroblasts in suspension culture after transfer directly to choline-free medium without a rinse was reduced to zero after 72 h. Monolayer cultures similarly treated multiplied at control rates for 96 h; one rinse of the latter prior to incubation in choline-free medium reduced growth at 48 h to 75% of control cells. A decrease in the size of cells in these rinsed monolayer cultures was apparent within 12 h, and preceded any changes in cell lipid composition; further decreases in cell size were observed at 24 and 48 h. After 24 h in choline-free medium the percent phosphatidyl choline had decreased only slightly and even at 48 h remained at 72 % of the value of control cells. At this time, however, there was a 50% decrease in the total phospholipid (PL) content of the cells, and a coincident 5–10-fold increase in the amount of triglyceride. There was no adaptive increase in the other principal PL classes. Changes in the ultrastructure of mitochondria were observed as early as 24 h; after 48 h without choline there was a decrease in the relative mitochondrial volume to 50% of control values. Alterations in the adhesive properties of choline-deprived cells may be related to altered lipid content or composition of the cell surface.  相似文献   

18.
Enriched protein diet-modified ghrelin expression and secretion in rats   总被引:5,自引:0,他引:5  
Gastrointestinal (GI) integrity and function are regulated by nutrition and growth factors. The discovery of ghrelin, a natural growth hormone (GH) secretagogue produced by the gastrointestinal (GI) tract, is a potential link between diet and growth signals. The aim of this study was to evaluate macronutrient effect on ghrelin expression and secretion in addition to some possible function in intestinal trophic status. Wistar rats were fed a high-carbohydrate, high-protein (HP), high-fat or standard (St) diet. Animals received the same daily food volume and caloric intake. After 7 days, animals were fasted for 24 h and blood and tissue samples were obtained just before feeding or at 2 or 6 h after feeding. Fasting high-protein-fed rats had higher ghrelin plasma levels than with rats fed the high-carbohydrate, high-fat or standard diets. Two-hours after refeeding, ghrelin plasma levels had decreased in all groups with a slight recovery at 6 h after refeeding, except in the high-protein group. Ghrelin plasma levels in rats fed with the high-protein diet correlated negatively with their GH and insulin-like growth factor 1 (IGF-1) plasma concentrations which were also the lowest among the study groups. In conclusion, ghrelin secretion was nutritionally manipulated because a protein-enriched diet increased its levels.  相似文献   

19.
Melanoma is the most dangerous type of skin cancer due to the occurrence of metastases. This work is aimed at studying the effects of the insertion of palmitic and oleic acid chain into monastrol in the melanoma cell line, B16F10. Cells were treated with monastrol, palmitic-monastrol or oleic-monastrol for periods of 0, 24, 48 and 72 h, and the cytotoxic effect was observed for palmitic-monastrol and oleic-monastrol after 24 h. For monastrol the effects were observed in 48 h on B16F10 cells, and in 24 h for a non-tumour cell line, melan-a. In this cell line, fatty-monastrol derivatives were cytotoxic after 24 h of exposure in the same concentrations as B16F10. However, oleic-monastrol inhibited cell growth at 20 µM only after 72 h, in contrast to the B16F10 cell line, in which oleic-monastrol inhibited cell growth at 48 h, showing that at least in this structural modification, melan-a was less sensitive than B16F10. The ability of compounds to induce apoptosis and/or necrosis was measured, and it was observed that monastrol induces apoptosis within 24 h. However, the cells treated with fatty-monastrol derivatives did not remain adhered on the well plate after 3 h of treatment. At this time point, these cells still emitted fluorescence indicating viable cells, suggesting a possible effect of palmitic- and oleic-monastrol in the adhesion proteins found on the cell membrane.  相似文献   

20.
Antioxidant defences and induced oxidative stress tissue damage of the blue shrimp Litopenaeus stylirostris, under challenge with Vibrio nigripulchritudo, were investigated for a 72-h period. For this purpose, L. stylirostris were first infected by immersion with pathogenic V. nigripulchritudo strain SFn1 and then antioxidant defences: superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (Gpx), Total antioxidant status (TAS), glutathiones and induced tissue damage (MDA and carbonyl proteins) were determined in the digestive gland at 0, 12, 24, 48 and 72 h post-infection (h.p.i.). In the meantime, TAS was also measured in the blood. Infection level of the shrimps during the challenge was followed by determining V. nigripulchritudo prevalence and load in the haemolymph of the shrimps. Changes in all these parameters during the 72-h.p.i. period were recorded for control shrimps and shrimps previously fed for one month with probiotic Pediococcus acidilactici MA18/5M at 107 CFU g?1 of feed.Our results showed that immersion with V. nigripulchritudo led to maximal infection level in the haemolymph at 24 h.p.i. preceding the mortality peak recorded at 48 h.p.i. Significant decreases in the antioxidant defences were detected from 24 h.p.i. and beyond that time infection leaded to increases in oxidative stress level and tissue damage. Compared to control group, shrimps fed the probiotic diet showed lower infection (20% instead of 45% at 24 h.p.i. in the control group) and mortality (25% instead of 41.7% in the control group) levels. Moreover, infected shrimp fed the probiotic compared to uninfected control shrimps exhibited very similar antioxidant status and oxidative stress level. Compared to the infected control group, shrimps fed the probiotic sustained higher antioxidant defences and lower oxidative stress level.This study shows that bacterial infection leads to oxidative stress in L. stylirostris and highlighted a beneficial effect of P. acidilactici, suggesting both a competitive exclusion effect leading to a reduction of the infection level and/or an enhancement of the antioxidant status of the shrimps.  相似文献   

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