Phenotypic variations in osmotic lysis of Sahel goat erythrocytes in non-ionic glucose media

2016 ◽  
Vol 27 (2) ◽  
Author(s):  
Nanacha Afifi Igbokwe ◽  
Ikechukwu Onyebuchi Igbokwe
Keyword(s):  
Glucose Transporter ◽  
Osmotic Fragility ◽  
Late Pregnancy ◽  
Transporter Protein ◽  
Extracellular Glucose ◽  
Pregnancy And Lactation ◽  
Osmotic Lysis ◽  
Sahel Goat ◽  
In Vitro Interaction

AbstractErythrocyte osmotic lysis in deionised glucose media is regulated by glucose influx, cation efflux, and changes in cell volume after water diffusion. Transmembrane fluxes may be affected by varied expression of glucose transporter protein and susceptibility of membrane proteins to glucose-induced glycosylation and oxidation in various physiologic states.Variations in haemolysis of Sahel goat erythrocytes after incubation in hyposmotic non-ionic glucose media, associated with sex, age, late pregnancy, and lactation, were investigated.The osmotic fragility curve in glucose media was sigmoidal with erythrocytes from goats in late pregnancy (PRE) or lactation (LAC) or from kid (KGT) or middle-aged (MGT) goats. Non-sigmoidal phenotype occurred in yearlings (YGT) and old (OGT) goats. The composite fragility phenotype for males and non-pregnant dry (NPD) females was non-sigmoidal. Erythrocytes with non-sigmoidal curves were more stable than those with sigmoidal curves because of inflectional shift of the curve to the left. Erythrocytes tended to be more fragile with male than female sex, KGT and MGT than YGT and OGT, and LAC and PRE than NPD. Thus, sex, age, pregnancy, and lactation affected the haemolytic pattern of goat erythrocytes in glucose media.The physiologic state of the goat affected the in vitro interaction of glucose with erythrocytes, causing variations in osmotic stability with variants of fragility phenotype. Variations in the effect of high extracellular glucose concentrations on the functions of membrane-associated glucose transporter, aquaporins, and the cation cotransporter were presumed to be relevant in regulating the physical properties of goat erythrocytes under osmotic stress.

Phenotypic homogeneity with minor deviance in osmotic fragility of Sahel goat erythrocytes in non-ionic sucrose media during various physiologic states

2016 ◽  
Vol 27 (6) ◽  
Author(s):  
Nanacha Afifi Igbokwe ◽  
Ikechukwu Onyebuchi Igbokwe
Keyword(s):  
Osmotic Fragility ◽  
Late Pregnancy ◽  
Erythrocyte Osmotic Fragility ◽  
Osmotic Water ◽  
Sigmoidal Curve ◽  
Osmotic Lysis ◽  
Osmotic Behaviour ◽  
The Mean ◽  
A Minor ◽  
Sahel Goat

AbstractBackground:Erythrocyte swelling in non-ionic sucrose media and the subsequent osmotic lysis are influenced by mechanisms of regulatory volume adjustment and osmotic water permeability. Kinetics of transmembrane water and ion fluxes in varied physiologic states may determine the phenotype of erythrocyte osmotic fragility (EOF) and affect estimates of EOF.Methods:Effects of sex, age, late pregnancy (third trimester) and lactation on the haemolysis of Sahel goat erythrocytes incubated in a series of hyposmotic non-ionic sucrose media were investigated.Results:The fragiligram was sigmoidal in 72 (97%) out of 74 goats. Two male (3%) goats with low and high extreme median erythrocyte fragilities (MEF), had non-sigmoidal curves. The mean fragilities at osmolarities of 30–300 mosmol/L of sucrose and the mean osmolarities responsible for 10%–90% haemolysis (CHConclusions:Physiologic states of the goat did not affect EOF phenotype in non-ionic sucrose media. Sigmoidal fragility phenotype seemed to be homogeneously conserved by osmoregulatory mechanisms not partitioned by sex, age, late pregnancy or lactation, but a minor non-sigmoidal curve might have occurred due to altered erythrocyte osmotic behaviour that would require further investigation.

Influence of extracellular media’s ionic strength on the osmotic stability of Sahel goat erythrocytes

2015 ◽  
Vol 26 (2) ◽  
Author(s):  
Nanacha Afifi Igbokwe ◽  
Ikechukwu Onyebuchi Igbokwe
Keyword(s):  
Ionic Strength ◽  
Osmotic Fragility ◽  
Ionic Concentration ◽  
Fragility Curve ◽  
Osmotic Resistance ◽  
Volume Changes ◽  
Erythrocyte Osmotic Fragility ◽  
Osmotic Stability ◽  
Osmotic Lysis ◽  
Sahel Goat

AbstractHeparinised blood was exposed to osmotic lysis in hypotonic buffered saline to evaluate erythrocyte membrane stability. When KThe erythrocyte osmotic fragility curve in saline was hyperbolic even when the ionic concentration was reduced by 50% with saccharides. Haemolysis was higher with EDTA than heparinised blood at saline concentrations of 90 and 150–180 mosmol/L. The fragility curve was sigmoidal and shifted to the left when saline was completely substituted with a saccharide. The non-ionic saccharides increased erythrocyte osmotic resistance linearly (r=0.88; p<0.02) from median to minimal hyposmolarities (150–300 mosmol/L) and reduced the osmolyte concentration at median fragility by 36%. No effect occurred at <30–120 mosmol/L and >90% fragility; and saccharide concentrations were almost non-lytic at comparable saline concentrations evoking <10% haemolysis. Fragilities were neither affected by period (30–60 min) of incubation nor the type of saccharide used.In this study, the variation in osmotic stability of caprine erythrocytes was linked to ionic strength of the suspending extracellular media which seemed to exert an influence through transmembrane ion fluxes and regulatory volume changes in erythrocytes.

Effects of ovariectomy and exercise training on muscle GLUT-4 content and glucose metabolism in rats

1996 ◽  
Vol 80 (5) ◽  
pp. 1605-1611 ◽  
Author(s):  
P. A. Hansen ◽  
T. J. McCarthy ◽  
E. N. Pasia ◽  
R. J. Spina ◽  
E. A. Gulve
Keyword(s):  
Skeletal Muscle ◽  
Glucose Transport ◽  
Glucose Transporter ◽  
Hormone Deficiency ◽  
Ovarian Hormone ◽  
Transporter Protein ◽  
Glut 4 ◽  
Female Wistar Rats ◽  
Flexor Digitorum Brevis

The present study examined the effects of 6 wk of ovarian endocrine deficiency on skeletal muscle GLUT-4 glucose transporter protein and glucose transport activity in sedentary and endurance-trained rats. Female Wistar rats (10 wk old) underwent bilateral ovariectomy (OVX) or sham surgery followed by a 5-wk swim-training protocol. OVX resulted in no significant changes in glycogen or GLUT-4 glucose transporter concentration in the soleus, epitrochlearis, or flexor digitorum brevis (FDB) muscles or in basal and maximally insulin-stimulated 2-deoxy-D-[1,2-3H]glucose (2-[3H]DG) transport in the soleus or epitrochlearis, suggesting that moderate-duration ovarian hormone deficiency does not significantly impair insulin action in skeletal muscle. In contrast, OVX decreased the maximal activation of 2-[3H]DG transport in the FDB by in vitro electrical stimulation. OVX had no significant effect on the training-induced changes in oxidative enzyme activities, GLUT-4 protein expression, glycogen content, or insulin-stimulated 2-[3H]DG transport in the soleus or epitrochlearis. These findings provide the first evidence that ovarian hormone deficiency decreases contraction-stimulated glucose transport in skeletal muscle.

EFFECT OF PERINATAL HYPOTHYROIDISM ON PITUITARY SECRETION OF GROWTH HORMONE AND PROLACTIN IN RATS

1974 ◽  
Vol 62 (2) ◽  
pp. 213-223 ◽  
Author(s):  
SAKAE KIKUYAMA ◽  
HIROSHI NAGASAWA ◽  
REIKO YANAI ◽  
KOREHITO YAMANOUCHI
Keyword(s):  
Growth Hormone ◽  
Late Pregnancy ◽  
Gonadal Development ◽  
Sprague Dawley ◽  
Sprague Dawley Rats ◽  
Pregnancy And Lactation ◽  
Pituitary Glands ◽  
Pituitary Secretion ◽  
Prolactin Content

SUMMARY Female Sprague—Dawley rats were fed 6-propyl-2-thiouracil (PTU) in their diet during late pregnancy and lactation. The growth and gonadal development of their pups were inhibited and in females the day of vaginal opening and onset of oestrous cycles were delayed; thyroid glands were hypertrophied. Treatment of the pups with thyroxine largely reversed these changes. The effect on body weight persisted even after treatment with PTU had stopped. At 20 days of age, the anterior pituitary glands of the pups of PTU-treated mothers contained significantly less growth hormone (GH) and prolactin than those of normal pups of both sexes. These changes persisted at 60 days of age. If the pups of PTU-treated mothers were given thyroxine from day 1 to day 20 of age, pituitary GH and prolactin content on day 20 had returned towards normal values. Thyroid deficiency was found to suppress the synthesis and release of prolactin and the synthesis of GH by the pituitary in vitro. These findings suggest that thyroxine influenced the maturation of the pituitary directly and/or through the hypothalamus and that thyroxine deficiency in early life brought about persistent alteration of the pituitary secretion of GH and prolactin.

GLUT4-containing vesicles are released from membranes by phospholipase D cleavage of a GPI anchor

2002 ◽  
Vol 283 (2) ◽  
pp. E374-E382 ◽  
Author(s):  
Søren Kristiansen ◽  
Erik A. Richter
Keyword(s):  
Phospholipase D ◽  
Glucose Transporter ◽  
Glucose Transporter 4 ◽  
Protein Fractionation ◽  
Membrane Structures ◽  
Transporter Protein ◽  
Membrane Compartment ◽  
A Cell ◽  
Triton X

We have previously developed a cell-free assay from rat skeletal muscle that displayed in vitro glucose transporter 4 (GLUT4) transfer from large to small membrane structures by the addition of a cytosolic protein fraction. By combining protein fractionation and the in vitro GLUT4 transfer assay, we have purified a glycosylphosphatidylinositol (GPI) phospholipase D (PLD) that induces transfer of GLUT4 from small to large membranes. The in vitro GLUT4 transfer was activated and inhibited by suramin and 1,10-phenanthroline (an activator and an inhibitor of GPI-PLD activity, respectively). Furthermore, upon purfication of the GLUT4 transporter protein, the protein displayed an elution profile in which the molecular mass was related to the charge, suggesting the presence or absence of phosphate. Second, by photoaffinity labeling of the purified GLUT4 with 3-(trifluoromethyl)-3-(m-[125I]iodopenyl)diazirine, both labeled phosphatidylethanolamine and fatty acids (constituents of a GPI link) were recovered. Third, by using phase transition of Triton X-114, the purified GLUT4 was found to be partly detergent resistant, which is a known characteristic of GPI-linked proteins. Fourth, the purified GLUT4 protein was recognized by an antibody raised specifically against GPI links. In conclusion, GLUT4-containing vesicles may be released from a membrane compartment by action of a GPI-PLD.

The nutrition of grazing ewes during pregnancy and lactation: a comparison of alkane-based and chromium/in vitro-based estimates of herbage intake

2000 ◽  
Vol 51 (6) ◽  
pp. 765 ◽  
Author(s):  
H. Dove ◽  
M. Freer ◽  
J. Z. Foot
Keyword(s):  
Late Pregnancy ◽  
In Vitro Digestibility ◽  
Faecal Output ◽  
Stage Of Lactation ◽  
Herbage Intake ◽  
Faecal Samples ◽  
Pregnancy And Lactation ◽  
The Difference

The n-alkane and chromium/in vitro procedures for estimating herbage intake were compared in grazing ewes during late pregnancy, early lactation, and mid-lactation. To ensure differences in herbage intake, the ewes were grazed in 4 plots of phalaris-dominant pasture at 2 levels of stocking: 17.1 ewes/ha and 30.8 ewes/ha. To investigate whether either procedure for estimating herbage intake was influenced by supplement consumption, half of the ewes at each stocking level received 500 g/day air-dry of a pelletted supplement (1 : 1 milled oat grain : sunflower meal). Supplement intakes were estimated using tritiated gypsum as a marker. During intake measurement periods, ewes were dosed twice daily with both alkane capsules and capsules containing chromium sesquioxide. For the last 6 days of the 12-day dosing period, rectal faecal samples were taken twice daily, immediately before the dosing. Over these same periods, wether sheep fitted with faecal collection harnesses were similarly dosed and sampled, and their total faecal output collected to establish the faecal recovery of chromium and the alkanes. Herbage intakes were estimated using the C27/C28, C29/C28, C31/C32, and C33/C32 alkane pairs. Estimates of intake based on the shorter alkane pairs were lower than those estimated with the C33/C32 alkane pair, by amounts which differed between the periods. Evidence is presented that estimates based on the last pair of alkanes (C33/C32) are the most accurate and are also more accurate than those based on the chromium/in vitro procedure. The relationship between these 2 methods for estimating intake was different in mid-pregnancy compared with either stage of lactation. The consumption of supplement did not interfere with any of the methods for estimating herbage intake. Estimates of faecal output based on the use of chromium, C28 alkane, or C32 as an external marker were statistically identical, indicating that the difference between the 2 methods for estimating herbage intake was not related to a failure to accommodate the incomplete recovery of any of the markers used or to the failure of rectal grab samples to be representative of total faeces. Our results indicate that herbage collected by oesophageally fistulated (OF) sheep was representative of that grazed by the ewes and could thus be used to provide the herbage alkane data needed to estimate herbage intake by the alkane method. However, the in vitro digestibility values obtained from the OF samples did not represent the digestibilities actually occurring in vivo. This was the main cause of the observed difference between the 2 methods for estimating intake. Possible reasons for the differences between the in vitro and in vivo estimates of digestibility are discussed.

scholarly journals Protective role of tocopherol and ascorbic acid in taxol-treated human erythrocytes in vitro

2017 ◽  
Vol 1 ◽  
pp. 239784731770581
Author(s):  
Uzma Faridi ◽  
Fatema Alatawi ◽  
Magda Mostafa
Keyword(s):  
Ascorbic Acid ◽  
Superoxide Dismutase ◽  
Side Effects ◽  
Protective Effect ◽  
Haematological Toxicity ◽  
Osmotic Fragility ◽  
Natural Antioxidants ◽  
Drug Induced ◽  
Osmotic Lysis

Taxol is a well-known anticancer agent. It is used for the treatment of several kinds of cancer, including breast cancer, lung cancer and ovarian cancer. In spite of being a good chemotherapeutic agent, taxol has several side effects. Drug-induced haemolytic anaemia is one of the most common side effects of taxol. This study investigated the haemolytic effect of taxol on normal erythrocytes and the protective effect of natural antioxidants ascorbic acid and tocopherol in the presence of taxol. We evaluated the osmotic fragility and the activity of enzymes superoxide dismutase and catalase of erythrocytes in the presence of taxol alone and taxol in combination with tocopherol and ascorbic acid. Taxol-induced haematological perturbation significantly caused haemolysis and reduced the activities of superoxide dismutase and catalase in erythrocytes. The antioxidants tocopherol and ascorbic acid demonstrated a protective effect when added to taxol. The combination of tocopherol with taxol significantly protected the osmotic lysis of erythrocytes and increased the activities of superoxide dismutase but had less effect on catalase. Ascorbic acid showed significant protection of erythrocytes from osmotic lysis but didn’t show any significant effect on superoxide and catalase. The results suggest that both antioxidants, especially tocopherol, could exhibit a protective effect against taxol-induced haematological toxicity.

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