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Z Etzion

Publications and source records attributed to Z Etzion.

At least 19 recordsLinked to original sources

Normal Ca2+ extrusion by the Ca2+ pump of intact red blood cells exposed to high glucose concentrations.

The ATPase activity of the plasma membrane Ca2+ pump (PMCA) has been reported to be inhibited by exposure of red blood cell (RBC) PMCA preparations to high glucose concentrations. It has been claimed that this effect could have potential pathophysiological relevance in diabetes. To ascertain whether high glucose levels also affect PMCA transport function in intact RBCs, Ca2+ extrusion by the Ca2+-saturated pump [PMCA maximal velocity (V(max))] was measured in human and rat RBCs exposed to high glucose in vivo or in vitro. Preincubation of normal human RBCs in 30-100 mM glucose for up to 6 h had no effect on PMCA V(max). The mean V(max) of RBCs from 15 diabetic subjects of 12.9 +/- 0.7 mmol. 340 g Hb(-1). h(-1) was not significantly different from that of controls (14.3 +/- 0.5 mmol. 340 g Hb(-1). h(-1)). Similarly, the PMCA V(max) of RBCs from 11 streptozotocin-diabetic rats was not affected by plasma glucose levels more than three times normal for 6-8 wk. Thus exposure to high glucose concentrations does not affect the ability of intact RBCs to extrude Ca2+.

Animals↗

Identification and characterization of a newly recognized population of high-Na+, low-K+, low-density sickle and normal red cells.

We describe a population of sickle cell anemia red cells (SS RBCs) ( approximately 4%) and a smaller fraction of normal RBCs (<0.03%) that fail to dehydrate when permeabilized to K(+) with either valinomycin or elevated internal Ca(2+). The nonshrinking, valinomycin-resistant (val-res) fractions, first detected by flow cytometry of density-fractionated SS RBCs, constituted up to 60% of the lightest, reticulocyte-rich (R1) cell fraction, and progressively smaller portions of the slightly denser R2 cells and discocytes. R1 val-res RBCs had a mean cell hemoglobin concentration of approximately 21 g of Hb per dl, and many had an elongated shape like "irreversibly sickled cells," suggesting a dense SS cell origin. Of three possible explanations for val-res cells, failure of valinomycin to K(+)-permeabilize the cells, low co-ion permeability, or reduced driving K(+) gradient, the latter proved responsible: Both SS and normal val-res RBCs were consistently high-Na(+) and low-K(+), even when processed entirely in Na-free media. Ca(2+) + A23187-induced K(+)-permeabilization of SS R1 fractions revealed a similar fraction of cal-res cells, whose (86)Rb uptake showed both high Na/K pump and leak fluxes. val-res/cal-res RBCs might represent either a distinct erythroid genealogy, or an "end-stage" of normal and SS RBCs. This paper focuses on the discovery, basic characterization, and exclusion of artifactual origin of this RBC fraction. Many future studies will be needed to clarify their mechanism of generation and full pathophysiological significance.

Anemia, Sickle Cell↗

K(86Rb) transport heterogeneity in the low-density fraction of sickle cell anemia red blood cells.

Previous studies have suggested ion transport heterogeneity among sickle cell anemia (SS) reticulocytes that could influence their dehydration susceptibility. We examined Ca2(+)-independent K transport in the lowest density (F1), reticulocyte-rich SS cells, measuring the effects of acidification, ouabain, and bumetanide on their unidirectional K(86Rb) fluxes. Unlike those of normal red blood cells and SS discocytes, the SS-F1 K(86Rb) fluxes were highly nonlinear, with large 5-min flux components (previously unobserved) and a more gradual decline over 60 min. Analysis revealed two distinct K pools: a rapid-turnover pool in a small fraction of cells, whose major ouabain-resistant K(86Rb) transport path showed distinctive properties including inhibition by high concentrations of bumetanide (> or = 1 mM) and stimulation at pH 7.0, and another heterogeneous, relatively slow-turnover pool, in most of the F1 cells, whose main ouabain-resistant K(86Rb) path was insensitive to bumetanide but was stimulated at pH 7.0, which is consistent with heterogeneous expression of the acid-sensitive K-Cl cotransport and with both rapid and slower generation of dehydrated SS cells.

Anemia, Sickle Cell↗

The distribution of intracellular calcium chelator (fura-2) in a population of intact human red cells.

Using quantitative fluorescence microscopy of red cells loaded non-disruptively with 1-2.5 mmol/l cells of fura-2, we examined the distribution of the incorporated free chelator among and within individual cells. Cytoplasmic hemoglobin quenched the effective fluorescence yield of fura-2 by a factor of about 100. All red cells were found to fluoresce upon excitation at 380 nm, and the fluorescence intensities they emitted at 510 nm were approximately +/- 20% about the mean intensity, indicating a fairly uniform distribution of incorporated chelator among the cells. Red cells loaded with these high levels of fura-2 retained their biconcave shape, and a comparison between their transmission images at 415 nm and their fura-2 fluorescence images suggests that the concentration of fura-2 was also uniform throughout the cytosol. These results validate assumptions made in earlier experiments with non-fluorescent incorporated Ca2+ chelators, and demonstrate the feasibility of fura-2 and Ca2+ imaging of intact red cells, despite considerable quenching of probe fluorescence by hemoglobin.

Calcium↗

Effects of deoxygenation on active and passive Ca2+ transport and cytoplasmic Ca2+ buffering in normal human red cells.

1. The effects of deoxygenation on cytoplasmic Ca2+ buffering, saturated Ca2+ extrusion rate through the Ca2+ pump (Vmax), passive Ca2+ influx and physiological [Ca2+]i level were investigated in human red cells to assess whether or not their Ca2+ metabolism might be altered by deoxygenation in capillaries and venous circulation. 2. The study was performed in fresh human red cells maintained in a tonometer either fully oxygenated or deoxygenated. Cytoplasmic Ca2+ buffering was estimated from the equilibrium distribution of 45Ca2+ induced by the divalent cation ionophore A23187 and the Vmax of the Ca2+ pump was measured either by the Co(2+)-exposure method or following ionophore wash-out. The passive Ca2+ influx and physiological [Ca2+]i were determined in cells preloaded with the Ca2+ chelator benz-2 and resuspended in autologous plasma. 3. Deoxygenation increased the fraction of ionized Ca2+ in cell water by 34-74% and reduced the Vmax of the Ca2+ pump by 18-32%. 4. To elucidate whether or not these effects were secondary to deoxygenation-induced pH shifts, the effects of deoxygenation on cell and medium pH, and of pH on cytoplasmic Ca2+ binding and Ca2+ pump Vmax in oxygenated cells were examined in detail. 5. Deoxygenation generated large alkaline pH shifts that could be explained if the apparent isoelectric point (pI) of haemoglobin increased by 0.2-0.4 pH units in intact cells, consistently higher than the value of 0.15 reported for pure haemoglobin solutions. 6. In oxygenated cells, the fraction of ionized cell calcium, alpha, was little affected by pH within the 7.0-7.7 range. Ca2+ pump Vmax was maximal at a medium pH of about 7.55. Comparison between pH effects elicited by HCl-NaOH additions and by replacing Cl- with gluconate suggested that Vmax was inhibited by both internal acidification and external alkalinization. Since deoxygenation alkalinized cells and medium within a range stimulatory for Vmax, the inhibition observed was not due to pH. 7. There was no significant effect of deoxygenation on passive Ca2+ uptake, or steady-state physiological [Ca2+]i level. 8. The deoxygenation-induced reduction in Ca2+ binding capacity may result from the increased protonation of haemoglobin on deoxygenation and from binding of 2,3-diphosphoglyceric acid (2,3-DPG) and ATP to deoxyhaemoglobin; inhibition of the Ca2+ pump may result from shifts in the [Mg2+]i/[ATP]i ratio away from a near optimal stimulatory value in the oxygenated state.

Biological Transport↗

Effects of deoxygenation on active and passive Ca2+ transport and on the cytoplasmic Ca2+ levels of sickle cell anemia red cells.

Elevated [Ca2+]i in deoxygenated sickle cell anemia (SS) red cells (RBCs) could trigger a major dehydration pathway via the Ca(2+)-sensitive K+ channel. But apart from an increase in calcium permeability, the effects of deoxygenation on the Ca2+ metabolism of sickle cells have not been previously documented. With the application of 45Ca(2+)-tracer flux methods and the combined use of the ionophore A23187, Co2+ ions, and intracellular incorporation of the Ca2+ chelator benz-2, in density-fractionated SS RBCs, we show here for the first time that upon deoxygenation, the mean [Ca2+]i level of SS discocytes was significantly increased, two- to threefold, from a normal range of 9.4 to 11.4 nM in the oxygenated cells, to a range of 21.8 to 31.7 nM in the deoxygenated cells, closer to K+ channel activatory levels. Unlike normal RBCs, deoxygenated SS RBCs showed a two- to fourfold increase in pump-leak Ca2+ turnover. Deoxygenation of the SS RBCs reduced their Ca2+ pump Vmax, more so in reticulocyte- and discocyte-rich than in dense cell fractions, and decreased their cytoplasmic Ca2+ buffering. Analysis of these results suggests that both increased Ca2+ influx and reduced Ca2+ pump extrusion contribute to the [Ca2+]i elevation.

Anemia, Sickle Cell↗

Isolation and characterization of rhoptries of Plasmodium falciparum.

Rhoptries have been isolated from Plasmodium falciparum schizont-infected erythrocytes by isopycnic density centrifugation. Gradient fractions were analyzed by immunoblotting with antibodies against two polypeptides of 140 and 110 kDa, known to be components of the rhoptry. The proteins were present primarily in fractions with a density of 1.16 g ml-1. Electron microscopy of these fractions indicated they were enriched in rhoptries. For the most part, the isolated organelle retained in situ morphology, although some rhoptries were distorted, indicating the structure of some of the organelles is not rigid. Electrophoretic analysis of the rhoptry fractions indicated the presence of a number of proteins, many of which have not been identified to date. Properties of proteins in the isolated rhoptry were examined using the 140 and 110 kDa proteins as representative markers. Both proteins are present in a complex with a 130-kDa protein, as all three co-immunoprecipitate. At the late schizont stage, the rhoptry proteins are present in two distinct forms; a soluble form with an Mr of 480 000 which would correspond to a single copy of the 140/130/110 kDa complex and a form that can be sedimented at 130 000 x g. Properties of the sedimentable form suggest that the proteins are included in structures that resemble membranes. Ionic detergents were required to solubilize the proteins while high concentrations of NaCl and Na2CO3 resulted in only partial solubilization. Furthermore, treatment of disrupted rhoptries with phospholipase A and C resulted in the release of proteins into the soluble form.

Animals↗

Erythrocyte invasion by two Plasmodium falciparum isolates differing in sialic acid dependency in the presence of glycophorin A antibodies.

Merozoites of Plasmodium falciparum depend on glycophorins for invasion into human erythrocytes, although this dependency varies between different geographic isolates of the species. The FCR-3 (Gambia) isolate appears to be fully dependent on the N-acetylneuraminic acid (NeuNAc) residues of the O-linked tetrasaccharide of glycophorin for invasion. Invasion of the CDC-1 (Honduras) isolate into neuraminidase treated erythrocytes is 50% of that into normal erythrocytes. This and additional results suggest that this isolate is not fully dependent on the O-linked saccharides of glycophorin. In the present study, invasion of CDC-1 and FCR-3 isolates into erythrocytes was examined in the presence of Fab fragments of monoclonal antibodies directed against different domains of glycophorin. Fab fragments directed against the carbohydrate domain inhibited invasion of both isolates but inhibited invasion by the FCR-3 isolate more than CDC-1 isolate. The reactivity of monoclonal antibodies (Mabs) directed against the carbohydrate domain was dependent on the NeuNAc residues as binding was abolished in neuraminidase treated erythrocytes. Mabs directed against the peptide domain of glycophorin A did not significantly inhibit invasion by either isolate. These results are consistent with other findings that the CDC-1 isolate is not fully dependent on the carbohydrate domain of glycophorin for invasion.

Animals↗

Localization of a parasite encoded protein to erythrocyte cytoplasmic vesicles of Plasmodium falciparum-infected cells.

Intracellular development of the malarial parasite results in substantial modifications of the membrane and cytoskeleton of the erythrocyte host cell. Two related Plasmodium falciparum-encoded proteins of 50 kDa and 43 kDa (Pf 50/43), identified by reactivity with a single mAb, were demonstrated to be localized to the erythrocyte cytoplasm of parasite-infected cells. Immunofluorescence and immunoelectron microscopy using mAb.7E11 demonstrated the Pf 50/43 is localized in the membrane of the vesicles in the erythrocyte cytoplasm, vesicles which correspond to Maurer's clefts. Solubility properties of the proteins suggest they are integral membrane proteins. By immunofluorescence, Pf 50/43 is shown to colocalize with actin which has a highly modified organization in the infected erythrocyte. Pf 50/43 is located exclusively in the vesicles, is not transported to the erythrocyte membrane or secreted. It is proposed the vesicles may play a role in transport of molecules across the erythrocyte cytoplasm, between the parasite and the external erythrocyte plasma membrane.

Animals↗

Metabolic effects in rats drinking increasing concentrations of sea-water.

Research on laboratory rats confirmed that drinking sea-water when dehydrated, was not beneficial and caused impaired renal function. When the concentration of sea-water in the drinking water is gradually increased there is a gradual increase in water uptake and corresponding urine excretion. At 50% sea-water the maximum uptake and excretion is reached. Following this there is a decline in appetite, water uptake and urine secretion. When on 100% sea-water, the creatinine clearances were greater than on tap water, while urine/plasma osmolalities (U/P) averaged 7. The only higher U/P was found in animals drinking sea-water when dehydrated, i.e. a U/P of 11. The urea metabolism appears to be suited to either the need to conserve body water, up to 50% sea-water, or to guarantee an adequate urine production, from 50% sea-water to pure sea-water. It is suggested that when a man is stranded at sea it is not advisable to drink all the fresh water and then be compelled to drink sea-water when dehydrated. It is better to slowly increase the sea-water uptake. This will prolong the time before sea-water needs to be drunk and result in only minor metabolic changes. Return to fresh water will be followed by an immediate return to normal homeostasis.

Animals↗

Uptake, distribution, and excretion of 31silicon in normal rats.

This study examines the uptake, distribution, and excretion of 31-labeled silicic acid in rat tissues at 1, 2, and 4 h after intracardiac injection of 31Si(OH)4. Plasma levels of 31Si decrease rapidly from 0.71 +/- 0.04% at 1 h to 0.07 +/- 0.06% of the dose administered per milliliter at 4 h. 31Si in plasma was found to be virtually entirely nonprotein bound. Kidney, liver, and lung accumulated the greatest amounts of 31Si per gram of wet weight, with concentrations at 4 h suggesting both relatively avid uptake and retention. Bone, skin, spleen, muscle, and testes also accumulated 31Si, but the levels were considerably lower than the aforementioned organs. Brain, however, contained negligible concentrations of 31Si throughout the study, indicating active exclusion by the blood-brain barrier. The major portion of the administered 31Si, 77 +/- 12%, was recovered in the urine within 4 h.

Animals↗

Camels' milk: for drinking only?

Camels' milk, women's milk and cows' milk were kept at 30 degrees C and refrigerated at 4 degrees C. This explains the necessity to immediately freeze milk if it needs to be kept even for a few days. Cows' milk remained good for days if stirred and then turned sour, enabling the making of cheeses and butter. Camels' milk did not sour at 4 degrees C for up to 3 months. This means that camels' milk is mainly good only for drinking, as was promised to this animal by the Prophet.

Animals↗

Tritiated water metabolism during dehydration and rehydration in the camel.

The metabolism of tritiated water in the camel was compared in two 10-day periods, one when water was readily available and the second during dehydration. There was a radically depressed metabolism after 2 days of dehydration. Two other experiments examined the absorption rate of drinking water. In one experiment drinking water was labeled with tritium, and in the second experiment dilution of tritium-labeled blood was examined. In both experiments there was a rapid uptake and dilution of the blood, which continued for 4 h. Following this only slight changes were observed up to 24 h after drinking. The results are in accord with other data showing changes in erythrocyte shape 4 h after rapid rehydration. It is concluded that there is a rapid absorption of water in the rehydrating camel.

Absorption↗

Growth and bone mineralisation as affected by dietary calcium, phytic acid and vitamin D.

1. Rats were fed various diets ranging from the normal chow, pure flour containing large amounts of phytic acid, Ca-enriched flour and mixtures of flour and normal food with various levels of calcium. 2. It was found that the animals eating the pure flour grew less and were smaller. 3. They suffered from hypocalcemia and had low plasma alkaline phosphatase and 25-HCC-vitamin D3 levels. 4. These animals had rib-cage deformities. 5. Additional calcium in the flour improved the animals' growth and calcification. 6. The mixed food did not greatly affect the animals and additional calcium did not improve growth or bone mineralisation. 7. The Bedouin eat large amounts of unleavened bread containing large amounts of phytates. 8. It is concluded that uptake of large amounts of phytates by the Bedouin eating unleavened bread is due to the flour and that the clinical manifestations are a direct result of the flour and not the lack of vitamin D due to covering the skin from sunlight.

Animals↗

Renal involvement following near-drowning in the sea.

1. The metabolic changes in rats following introduction of sea water directly into the stomach were studied. The results were compared with those of a human case report of acute renal failure following swallowing of sea water. 2. In both rats and man, acute renal failure appeared within the first 24 hr. A slight improvement was found five days later. 3. There was an acute hypocalcemia. This was supposedly due to the large magnesium content of the sea water. 4. It is probable that water entering the alimentary canal, dry drowning, affects the kidneys by the initial flow of water to the intestines. This results in hemo-concentration and reduced kidney function. The abnormality is a transient one and could be due to a partial anoxia due to shrinking of the erythrocytes. 5. It is concluded that an apparently uneventful swallowing of sea water can turn into secondary drowning with transient acute renal failure.

Animals↗

Cooking-smoke as a factor affecting thyroid metabolism and oxygen transport capability.

1. Young Bedouin women found to be prone to illnesses of the thyroid gland and lungs. It was surmised that the cause of both complaints had the same origin: the smoke from cooking over a fire. 2. Laboratory rats were placed in the cooking space in a Bedouin encampment for 2 months. These rats (termed "smoke" animals) were smaller and less active than normal siblings. 3. The smoke animals had increased secretion of thyroid hormones and TSH and the thyroid glands were enlarged, when compared to body weight. 4. Histology of the thyroid showed areas of atrophy, areas of hypertrophy and cysts. There was increased erythropoiesis, allowing for enhanced oxygen transport capability. 5. It is concluded that the smoke, directly, or indirectly by hampering normal lung function, caused a reaction in the thyroid gland, which in turn led to increased production of hemoglobin, probably to guarantee sufficient oxygen for basic cell metabolism.

Animals↗

Hormonal and behavioural patterns in the male camel (Camelus dromedarius).

Androgen levels in blood and the behaviour of male camels were determined during 1 year. In the months January to April large amounts of androgens were found in blood serum (approximately 30 ng/ml compared with approximately 2 ng/ml throughout the remaining months) as well as in a secretion from glands on the back of the neck (36 ng/ml). During this period when hormone levels were high the behaviour of the camels changed: they became unmanageable, blew-out a palatal flap from the mouth, vocalized, and spread urine over their backs with the aid of their tails. The changes in the sexual cycle of the male camels corresponded with the oestrous period in female camels.

Androgens↗