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[Anti-oxidative effects of proanthocyanidins in mice induced by D-galactose].

The anti-oxidative effect of proanthocyanidins(PC) was evaluated in D-galactose-induced murine model. Male KUNMING mice were divided into different groups at random. The mice were induced by subcutaneous injection of D-galactose on the back of mice daily for 60 days and simultaneously PC and VE were administered to the mice by oral feeding. Results indicate that the lipid peroxide in blood, liver and brain significantly increased respectively because of the D-galactose injection, when compared to the control. The activity of SOD in RBC and liver significantly decreased while that of GSH-PX in blood and liver decreased. MAO-B in brain was significantly increased while MAO-B in liver doesn't change significantly. Oral feeding PC markedly decreased the formation of MDA in blood, liver and brain, increased the activities of SOD in blood and liver and decreased the activity of MAO-B in brain. Vitamin E also counteract the oxidative stress induced by D-galactose. In summary, treatment with PC at all the three tested doses were effective in exerting a protective effect against oxidative stress.

Aging↗

[Urethrosonocystography with galactose in the diagnosis and follow-up of pediatric patients with vesicoureteral reflux].

INTRODUCTION AND OBJECTIVES: Since early 80's, attempts to detect Vesicoureteric Reflux (VUR) with Ultrasound (US) contrast medium have been made to study the urinary tract during voiding. The galactose-based agents are comparable in the diagnostic range with the standard fluoroscopic cystography, providing high values of sensitivity and specificity. The purpose of our work is to show our experience during last three years with the urethrosonocystography with echo contrast in the diagnosis and follow-up of pediatric patients with urological malformations. PATIENTS AND METHODS: 442 patients were included in our study (aged 1 day to 13 years; 202 males and 240 females). All cases were referred for investigation of VUR based on previous criteria of urinary tract infection, VUR follow-up, dilated urinary tract, post endoscopic treatment, surgical treatment follow-up and others. After transurethral catheterisation the bladder was filled with US galactose-based contrast medium (Levovist). VUR was diagnosed when micro bubbles appeared in ureter or pelvicalyceal system and was graded according to the International Reflux Study Committee. RESULTS: Of the 442 patients evaluated with echo enhanced urethrosonocystography, 227 were screening cases for discard VUR. Of these patients, only 58 were diagnosed as having VUR. In 165 cases the technique was indicated as follow-up. Two patients with spina bifida developed candiduria after the sonographic cystogram. In 6 patients with dilated urinary tract without reflux, galactose was detected in renal pelvis 6 months after the procedure. CONCLUSIONS: In our experience urethrocystosonography with galactose-based US medium agents is a more sensitive method than standard Voiding Cystourethrography (VCUG) for detecting VUR in pediatric patients. US cystography is an attractive technique which involves no ionising radiation and is usually well tolerated by the young. This procedure should be considered as a routine diagnostic work-up for detecting VUR in pediatrics.

Child↗

Alterations in lens permeability during galactose cataract development in rat.

Lens permeability has been shown to be compromised during galactose-induced cataract development in rats. Recent studies have demonstrated permeability and diffusion pathways as well as endocytotic activity in normal lenses of different species using tracers of different molecular weights. We investigated the permeability and diffusion of tracers in normal rat lenses and in lenses during cataract development using three different molecular weight tracers, lanthanum nitrate (LN, MW 430), ruthenium red (RR, MW 858.5) and horseradish peroxidase (HRP, MW 40.000) for this study. Sprague Dawley rats weighing 50gms were fed Purina Rat Chow with or without galactose. Our results, based on transmission electron microscopy, show that all tracers penetrated through the capsule and the basal portion of the intercellular spaces. While the diffusion of RR was restricted to the epithelial cell layer, LN and HRP were observed in intercellular spaces in cortical fiber cells. In the HRP "washout" studies, HRP could be readily removed from the intercellular spaces in the basal region in the epithelial cell layer. This and other observations suggest the presence of a barrier (tight-junction) in the apical region. Our studies also suggest an influx of tracers in the lens, specifically LN and HRP, through the equatorial region. The permeability of the tracers increased and endocytotic vesicles with tracers were often found associated with the lateral and basal membranes of epithelial cells in the galactose-fed rats at the precataractous and mature cataractous stages. This study provides further support for the presence of a tight-junction between epithelial cells and indicates the movement of tracers through the equatorial region. Moreover, it confirms previous observations that indicated alterations in lens permeability during galactose cataractogenesis.

Animals↗

[Effect of tetramethylpyrazine on learning, memory and cholinergic system in D-galactose-lesioned mice].

OBJECTIVE: To explore the effect of tetramethylpyrazine on learning, memory, and cholinergic system in D-galactose-lesioned mice. METHODS: C57BL/6J mice were given subcutaneous injection of 2% D-galactose for 40 days (100 mg.kg-1.d-1). Normal saline, tetramethylpyrazine (TMP) and Huperzine A (HupA) were given respectively by intragastric administration in different study groups from the third week on. Learning and memory ability were tested by Morris water maze for 5 days at the sixth week. Acetylcholinesterase (AchE) activity, the binding sites (Bmax) and the affinity (KD) of M-cholinergic receptor were determined. RESULTS: The learning and memory dysfunction, with lowered AchE activity and M-cholinergic receptor binding sites were found in the model group as compared with the normal control group. The tetramethylpyrazine, especially at the dose of 100 mg.kg-1.d-1, could markedly attenuate cognitive dysfunction, while elevate the lowered AchE activity (P < 0.05) and M-cholinergic receptor binding sites (P < 0.005) in the cerebral cortex of mice treated with D-galactose. CONCLUSIONS: The tetramethylpyrazine can significantly improve central cholinergic system function, and thus enhance the learning and memory ability in D-galactose-lesioned mice.

Acetylcholinesterase↗

[Effects of puerarin on fatty superoxide in aged mice induced by D-galactose].

OBJECTIVE: To study the effect of puerarin on fatty superoxide of aged mice induced by D-galactose. METHOD: The aged mice were induced by s.c. 0.12 g.kg-1 D-galactose for 6 weeks. Meanwhile, they were treated with three doses of puerarin once a day for 6 weeks. Then the spontaneous behavior was tested in the aged mice using open field at the next day after the last treatment. And the activity of SOD, the contents of MDA and lipofuscin in brain, liver, and serum were measured with Ultravioletray spectrometer and Fluorospectrophometer analysis system. RESULT: Compared with the D-galactose control group, puerarin of 50 mg.kg-1 and 100 mg.kg-1 puerarin significantly increased the spontaneous behavior, remarkably promoted the activity of SOD of brain, liver, and serum in the aged mice, and significantly decreased the contents of MDA and lipofuscin. CONCLUSION: Puerarin can improve the activity of antioxidase of the modelling aged-mice induced by D-galactose.

Aging↗

Plasma insulin, carbohydrate, and free fatty acid changes in newly born infants of diabetic and non-diabetic mothers after loading with glucose, fructose, and galactose.

Changes in the concentration of blood glucose, fructose and galactose and their disappearance rate, concentration of plasma insulin, free fatty acids, blood lactate and pyruvate after intravenous glucose, fructose and galactose loads (1 g/kg) were studied in 23 infants of insulin treated diabetic mothers (IDM). The control group consisted of 42 infants of healthy mothers (IHM). The disappearance rate of these monosaccharides was higher in IDM that in IHM (P less than 0.01). All monosaccharides enhanced plasma insulin levels, but the plasma insulin concentration varied considerably. The highest insulin response with difference between IDM and IHM occurred after loading with glucose. Fructose was the least effective insulin stimulator which did not increase glucose levels. All monosaccharides decreased free fatty acid levels. Lactate levels and lactate:pyruvate ratio in IHM were increased after fructose loading. The results of this study suggest that galactose is of some physiological importance for maintaining glucose levels and that the islet apparatus of newborns of diabetic mothers is less loaded with galactose than with glucose.

Blood Glucose↗

[The experimental study of Radix Puerariae inhibiting glycation in rats induced by D-galactose].

OBJECTIVE: To observe the effects of Radix Puerariae on protein glycation in model rats induced by D-galactose. METHODS: The model rats of protein glycatin were induced by intraperitoneal administration of D-galactose (150 mg/kg) for 8 weeks, and all rats were treated by Radix Puerariae (High dose 300 mg/kg, Middle dose 150 mg/kg, Low dose 75 mg/kg) for 6 weeks. The activity of aldose reductase in red blood cells, the content of fructosamine in serum, the amount of glycohaemoglobin and advanced glycation end-products, the level of insulin in serum, the activity of superoxide dismutase and the amount of maleic dialdehyde were measured. RESULTS: High dose and middle dose of Radix Puerariae could decrease the level of blood glucose and the activity of aldose reductase in red blood cells, inhibit the formation of glycation products significantly in model rats induced by D-galactose (P < 0.01), increase insulin sensitivity and activity of superoxide dismutase (P < 0.01) and decrease the amount of maleic dialdehyde (P < 0.01). CONCLUSION: Puerariae can significantly inhibit glycation reaction in rats induced by D-galactose.

Aldehyde Reductase↗

[Effects of green tea polyphenol on the behaviour of Alzheimer' s disease like mice induced by D-galactose and Abeta25-35].

OBJECTIVE: To study the effects of green tea polyphenol (GTP) on the behaviour of Alzheimer' s Disease like mice induced by D-galactose and Abeta25-35. METHODS: D-galactose (120 mg/kg) was intraperitoneally injected into mice for 12 weeks and Abeta25-35 (4nmol) was injected by icv to establish Alzheimer' s Disease like mice. The protective and therapeutic effects of GTP were determined by using water maze test, step down test, step through test and open field test. RESULTS: GTP ameliorated the deleterious effects of D-galactose and Abeta25-35, and thereby improved the animal' s learning and memory, prolonged latency time, and the error numbers were significantly reduced. At the same time the autonomic activities were significantly increased. CONCLUSION: GTP can improve the behaviour of Alzheimer' s Disease like mice induced by D-galactose and Abeta25-35.

Alzheimer Disease↗

Cumulative antioxidant defense against oxidative challenge in galactose-induced cataractogenesis in Wistar rats.

Natural dietary ingredients are known for their antioxidant activity. Of such, curcumin, the active principle of turmeric, at 0.01% in the diet proved as pro-oxidative in galactose-induced cataract in vivo. The purpose of this study was to investigate the effect of vitamin E (VE), a well-known antioxidant, in combination with curcumin on the onset and maturation of galactose induced cataract. Periodic slit-lamp microscope examination indicated that in combination with vitamin-E, 0.01% curcumin (G-IV) delayed the onset and maturation of galactose-induced cataract. Biochemical analyses revealed that combined treatment of 0.01% curcumin and vitamin-E diet exhibited an efficient antioxidant effect, as it inhibited lipid peroxidation and contributed to a distinct rise in reduced glutathione content. The results indicate that natural dietary ingredients are effective in combination rather than the individual administration as they are complementing each other in reducing the risk of galactose induced cataract.

Animals↗

[Improvement effects of puerarin on glycated brain damages in rats induced by D-galactose].

OBJECTIVE: To observe the improvement effects of puerarin on glycated brain damages in rat model induced by D-galactose. METHOD: The model rats of protein glycation were induced by intraperitoneal administration of D-galactose (150 mg x kg(-1) x d(-1)) for 8 weeks, and all rats were treated with puerarin (high dose 300 mg x kg(-1), middle dose 150 mg x kg(-1), low dose 75 mg x kg(-1)) for 6 weeks. The activity of aldose reductase in red blood cells, the amount of glycated products (fructosamine in serum, glycohaemoglobin, advanced glycation end-products) and AGEs in brain tissue, calcium ion in brain cells were measured. Moreover, mitochondria in brain hippocampus cells were observed under electronic microscope. RESULT: High dose and middle dose of puerarin can decrease the activity of aldose reductase in red blood cells (P < 0.01), and inhibit the formation of glycation products significantly in model rats induced by D-galactose (P < 0.01). Also, puerarin can decrease the content of AGEs in brain and the level of calcium ions in brain cells (P < 0.05, P < 0.01), and decrease lesions degree in mitochondria in brain hippocampus cells. CONCLUSION: Puerarin can produce the protective effects on glycated brain damages through inhibiting the glycation reaction in rats induced by D-galactose.

Aldehyde Reductase↗

Clearance of tissue plasminogen activator by mannose and galactose receptors in the liver.

The mechanism of uptake of radio-iodinated tissue plasminogen activator (125I-t-PA) was studied in rats. When trace amounts of 125I-t-PA were injected alone, the clearance followed a biphasic pattern in which 65% and 35% were cleared with alpha- and beta-kinetics (t1/2 (alpha) = 0.6 min, and t1/2 (beta) = 6.4 min), respectively. Co-injection with excess unlabelled t-PA or mannan changed the uptake kinetics to the monophasic beta-elimination pattern. Mannosylated albumin and ovalbumin, both of which bind to the hepatic mannose receptor, reduced the proportion of t-PA cleared with t1/2 (alpha) to 48% and 21%, respectively. A corresponding increase in the beta-elimination of t-PA was observed. The t1/2 (alpha) and t1/2 (beta) were unchanged. Studies on the clearance of 125I-ovalbumin also showed a biphasic elimination with an initial rapid phase, t1/2 (alpha), accounting for only 39% of the clearance of ovalbumin, as compared to 65% in the case of t-PA. Macromolecules with affinity for the galactose-receptor only, such as asialofetuin, or galactosylated albumin, did not significantly affect the clearance kinetics at the concentrations used. Asialoorosomucoid, which also carries galactosyl residues in the terminal position, reduced somewhat (from 65% to 48%) the proportion cleared with alpha-kinetics. Very high concentrations of galactose and N-acetyl-galactosamine, which are also known to compete for binding to the galactose receptor, lowered the proportion of t-PA cleared in the late beta-phase (reduced from 35% to 26% with galactose and to 19% with N-acetyl-galactosamine).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Red cell aging results in a change of cell surface carbohydrate epitopes allowing for recognition by galactose-specific receptors of rat liver macrophages.

Binding and uptake studies of in vitro aged or senescent rat erythrocytes by isolated rat liver macrophages suggest recognition by galactose-specific receptors on the cell surface of the macrophages. We analyzed carbohydrates exposed on old erythrocytes by plant lectins in an agglutination assay in comparison with freshly isolated untreated erythrocytes. Rat erythrocytes aged in vitro by storage are agglutinated by a panel of lectins that do not react with freshly isolated erythrocytes. Specificity of agglutination was shown by inhibition with monosaccharides. Antibodies eluted from senescent rat erythrocytes agglutinate in vitro aged as well as senescent rat erythrocytes, but not freshly isolated cells nor human erythrocytes. Galactose-specific lectins isolated from rat liver give similar results; they also agglutinate normal human erythrocytes. Agglutination by the liver lectin is inhibitable by galactose and N-acetylgalactosamine but not by N-acetylglucosamine or mannose. Furthermore, rat liver macrophages devoid of galactose-specific receptors show markedly reduced binding of senescent rat erythrocytes. We conclude that recognition of old rat erythrocytes is mediated by two systems: old erythrocytes expose different terminal sugar residues or a different arrangement of glycans when compared to young erythrocytes, rendering them recognizable by liver lectins and by autoantibodies.

Agglutination Tests↗

Elemental and structural studies of the rat galactose cataract.

A series of rat galactose lenses, from 1 to 20 days on the 50% galactose diet, were frozen in the whole eye, and fractured from pole to pole in the frozen state. Lyophilized half-lenses were prepared for analysis by energy dispersive spectrometry (EDS). Following elemental analysis, some specimens were embedded and sectioned for histological studies. Elemental X-ray maps, and/or profiles, were made for K, Na, Cl, Ca, P, and S. As early as two days on the galactose diet, a crescent-shaped region ("streak") of Cl, Na, and Ca gain, and K loss develops near the equatorial surface. Between this region and the equatorial surface are the nucleated differentiating fiber cells which maintain low Cl, Na, and Ca, and high K (viable equatorial zone, VEZ). With time the "streak" expands anteriorly, centrally and posteriorly, eventually (by 20 days) including most of the lens. The VEZ, including the epithelium, however, is non-reactive to the galactose diet, which is deleterious to the fully differentiated fiber cells. Eventually, the VEZ undergoes a characteristic morphological change, apparently due in part to changes in its physical environment.

Animals↗

Aldose reductase inhibitors and prevention of galactose cataracts in rats.

Our previous studies have shown that the aldose reductase inhibitor (ARI), sorbinil, prevents galactose-induced alterations and cataracts in rat lenses. We have now used sorbinil as well as another ARI, Eisai compound E-0722, to determine their potency in inhibiting aldose reductase- and galactose-induced alterations in lens morphology and Na+-K+-ATPase activity. Young Sprague Dawley rats were fed Purina Rat Chow plus 50% galactose, with or without 15 mg sorbinil, 0.15, 0.5 or 1.0 mg of E-0722/kg body weight per day. Controls were given Purina Rat Chow with or without ARIs. Lenses were studied for up to 60 days following the initiation of the diet using morphological, cytochemical and biochemical approaches to assess any alterations in the lens. While galactose-induced damage and cataracts were delayed by low doses (0.15 mg and 0.5 mg) of E-0722, they were completely prevented by the administration of 15 mg of sorbinil or 1 mg of E-0722/kg body weight per day. This study further showed that just 1 mg of E-0722 was more effective in preventing cataracts than 15 mg sorbinil. Thus it appeared that E-0722 was a more potent inhibitor of aldose reductase than sorbinil.

Aldehyde Reductase↗

Changes in alpha-lactalbumin, total lactose, UDP-galactose hydrolase and other factors in tammar wallaby (Macropus eugenii) milk during lactation.

alpha-Lactalbumin was isolated from milk of M. eugenii and its concentration in milk samples taken at various times during lactation (0-40 weeks post partum) was determined by single radial immunodiffusion using rabbit antiserum to the purified protein. The alpha-lactalbumin concentration remained almost constant throughout lactation even though the concentration of total lactose (free lactose plus lactose contained in oligosaccharides) fell to zero after 34 weeks post partum. This fall in lactose was accompanied by a rise in the free galactose and glucose concentrations and marked increases in UDP-galactose hydrolase, nucleotide pyrophosphatase, alkaline phosphatase and acid beta-galactosidase activities. It is suggested that the in vitro hydrolysis of UDP-galactose was due to nucleotide pyrophosphatase and that this enzyme may also play a role in vivo late in lactation by making UDP-galactose unavailable for the synthesis of lactose. Alternatively, lactose and lactose-containing oligosaccharides might be degraded by the acid beta-galactosidase during or after secretion.

Animals↗

Channeling of alpha-D-glucose 6-phosphate in tumoral islet cells exposed to D-galactose.

In human erythrocytes, in which the fractional turnover rate of glucose 6-phosphate is rather low, menadione increases to almost the same relative extent the oxidation of D-[U-14C]glucose and D-[U-14C]galactose. However, in pancreatic tumoral islet cells (RINm5F line), in which the fractional turnover rate of glucose 6-phosphate is considerably higher, menadione increases the oxidation of D-[1-14C]glucose but not that of D-[1-14C]galactose. These results suggest that alpha-D-glucose 6-phosphate generated from exogenous D-galactose is channeled preferentially into the glycolytic rather than pentose phosphate pathway. Such was no more the case, however, when the RINm5F cells were exposed simultaneously to both D-glucose and D-galactose.

Adenoma, Islet Cell↗

Kluyveromyces bulgaricus yeast lectins. Isolation of N-acetylglucosamine and galactose-specific lectins: their relation with flocculation.

Kluyveromyces bulgaricus is a yeast which, upon culture in a calcium-enriched glucose-peptone medium, flocculates. Its flocculation can be reversed by the addition of galactose. In this paper, it is shown that two lectins can be isolated either from the concentrated culture broth or from the supernatant of deflocculated cells suspended in galactose solution. The N-acetylglucosamine-specific lectin, at pH 7.4, agglutinates untreated sheep red blood cells, but agglutinates neither untreated rabbit red blood cells nor glutaraldehyde-fixed sheep or rabbit red blood cells. Conversely, at pH 4.5, this lectin agglutinates glutaraldehyde-fixed sheep red blood cells. The galactose-specific lectin, at pH 7.4, agglutinates both untreated and glutaraldehyde-fixed rabbit red blood cells but does not agglutinate untreated or glutaraldehyde-fixed sheep red blood cells. At pH 4.5, this lectin agglutinates both glutaraldehyde-fixed sheep and rabbit red blood cells and induces flocculation of deflocculated K. bulgaricus cells. In all cases, the agglutination and the flocculation induced by one of these two lectins were inhibited by free or conjugated N-acetyl-D-glucosamine or by free or conjugated D-galactose, respectively. No glycosylhydrolase activity could be detected in the purified lectins.

Acetylglucosamine↗

Sorbinil prevents the galactose-induced inhibition of prostaglandin synthesis in lens cells.

The relationship between microsomal prostaglandin biosynthesis, PGH synthase activity and cell morphology was investigated with cultured bovine lens epithelial cells under physiological and hypergalactosemic conditions. The rate of lens cell microsomal PGE2 generation fell from 5.3 to 2.2 pg micrograms protein-1 min-1; PGF2 alpha declined from 7.7 to 4.8 pg micrograms protein-1 min-1 within 20 hr of exposure to 40 mM galactose. The decreased PGE2 and PGF2 alpha biosynthetic capability was attributed to a reduction in PGH synthase activity which reduced to 62% of control (5.5 mM glucose) after a 20 hr exposure to galactose. Measurement of PGH synthase activity after 6 days of continuous exposure to galactose resulted in a further reduction to 55% of control. The diminution in microsomal prostagladin biosynthesis and decline in PGH synthase activity precluded ultrastructural alterations, such as large vacuole formation and severe autodegradation of organelles. The simultaneous introduction of sorbinil, an aldose reductase inhibitor, to the galactose medium not only prevented the decrease in microsomal prostaglandin biosynthesis and PGH synthase activity, but also the detrimental morphological complications. The lens epithelial cell system may provide a useful model for examining the biochemical and morphological-related consequences of sustained hypergalactosemia and their potential regulation by aldose reductase inhibitors.

Animals↗