Metabolic and functional characteristics of erythrocytes from Glycera and Noetia

• 1.1. Annelid and molluscan red blood cells (RBC) may de differentiated metabolically from vertebrate RBC by their increased permeability to substrate, their magnitude of amino acid catabolism and their higher aerobic metabolism.
• 2.2. At 22°C, Glycera and Noetia RBC oxidize glucose and glutamate to CO₂ without accumulation of either d- or l-lactate. By comparison, the oxidation of glutamate by rat and chicken RBC is negligible at this temperature despite its incorporation into the cells.
• 3.3. At 37°C, chicken RBC oxidize glutamate at a rate 4 times greater than at 22°C, with oxygen uptake still lower than that in Noetia RBC at 32°C. At 37°C, rat RBC do not increase their oxidation of glutamate above that at 22°C, but oxygen uptake increases to slightly more than half that of chicken RBC.
• 4.4. Our finclings indicate that RBC of these two invertebrate species have both a higher aerobic metabolism and lower anerobic capacity than vertebrate RBC.
• 5.5. Moreover, the annelid and molluscan RBC have a relatively lower activity of the pentose phosphate (PPO₄) pathway than vertebrate RBC, as evidenced by their higher thermal sensitivity of oxygen uptake and their higher *C₁O₂/*C₆O₂ isotope ratio.

     

Contrast in adenine uptake by chicken and rabbit erythrocytes in vitro

• 1.1. Relative to rabbit erythrocytes, chicken red blood cells exhibit a much greater capacity to utilize [³H]adenine for nucleotide synthesis in vitro, even at 5°C and in the absence of added inorganic phosphate.
• 2.2. This difference is largely due to a higher concentration of phosphoribosylpyrophosphate and greater activity of adenine phosphoribosyltransferase in the avian cells. lli]3. The capacity of avian erythrocytes for utilization of guanine and hypoxanthine is several fold less than that of adenine.
• 3.4. The data are consistent with lower activity for hypoxanthine/guanine phosphoribosyltransferase than for adenine phosphoribosyltransferase in intact chicken erythrocytes.
• 4.5. The results indicate that reutilization of adenine by chicken erythrocytes may be physiologically significant.

     

The effect of time on physiological changes in eel Anguilla anguilla,induced by lindane

• 1.1. Eel were exposed to a sublethal concentration of lindane (0.335 ppm) for 6, 12, 24, 48, 72 and 96 hr.
• 2.2. Concentrations of glycogen, glucose, lactate, pyruvate and lipids were determined in gill tissue after lindane exposure.
• 3.3. Gill glycogen descreased and glucose levels increased at 6 hr of treatment, lactate and pyruvate concentration increased between 6 and 48 hr. Total lipid values decreased between 6 and 24 hr; thereafter, the levels increased up to 72 hr of exposure.
• 4.4. Clear changes were found in all parameters tested in gill tissues. The observed effects of lindane on metabolism in fish are discussed in relation to acute stress syndrome.

     

Effect of ozone exposure on the compositions of gill and erythrocyte membrane lipids and proteins of Japanese charr (Salvelinus leucomaenis)

• 1.1. Compositions of lipids and proteins of erythrocytes (RBC) and gills from Japanese charr (Salvelinus leucomaenis) which were exposed to 0.4 and 0.7 ppm ozone for 30 min were compared with those of the control.
• 2.2. On exposure to ozone, both RBC and gill membrane phospholipid content, especially phosphatidylethanolamine (PE), dropped.
• 3.3. The decrease of PE was brought about by the decrease of docosahexaenoic acid content which comprised the major component of PE.
• 4.4. RBC membrane protein with 215 and 225 kDa, which is equivalent to cytoskeletal protein, selectively disappeared on exposure to ozone.

     

A comparative study of seafish erythrocytes and agglutinins from seaweeds

• 1.1. To date, only a few authors have assayed the agglutinic activity of marine algae against fish erythrocytes, and in these cases, mainly against freshwater fish.
• 2.2. For the first time, the hemagglutinic activity of 70 seaweeds (29 brown, 37 red and four green algae) against erythrocytes of 16 seaflsh species is reported.
• 3.3. The presence of agglutinins was demonstrated in 100% of algae assayed, against at least one of the different types of erythrocytes tested.
• 4.4. The results obtained confirm the presence of receptors for algae agglutinins on the surface of the erythrocytes of the fish studied.
• 5.5. This could be useful in establishing the origins of fish populations, as these serological differences could distinguish between populations of cultivated and wild fish.

     

Effects of dehydration and rehydration on the intravascular space in horses

• 1.1. The resistance of sub-tropical horses, and desert-dwelling horses to 72 hr dehydration/24 hr rehydration was investigated via changes in red cell parameters and plasma protein concentration.
• 2.2. Red cell count, haemoglobin and haematocrit increased up to 48 hr dehydration. Between 48 and 72 hr dehydration these parameters decreased, implying a fluid shift onto the intravascular space from the interstitium/hindgut. Most parameters had regained baseline values by 24 hr rehydration.
• 3.3. Mean cell volume, mean cell haemoglobin, mean cell haemoglobin concentration and total plasma protein were not significantly different between breeds at, or between most stages of hydration.
• 4.4. Protection of plasma volume during dehydration/rehydration was aided by maintaining intravascular protein (especially albumin) levels. Red cells were transiently dehydrated and overhydrated but resisted osmolysis.

     

Acute stress suppresses plasma estradiol levels in female alligators (Alligator mississippiensis)

• 1.1. Five adult, female alligators (Alligator mississippiensis) were captured at night during the breeding season, and a blood sample taken within 5 min of capture.
• 2.2. The alligators were physically restrained (tied to boards) and additional blood samples taken at 4, 8, 12, 16, 22, 28, 38, and 48 hr after capture. After the last blood sample was collected the animals were released.
• 3.3. Plasma estradiol-17β and corticosterone were measured by radioimmunoassay. Estradiol declined significantly from initial values by 22 hr post capture, but remained unchanged for 48 hr.
• 4.4. Plasma corticosterone rose from a mean of 0.8 ng/ml at capture to 12.6 ng/ml after 4 hr. Corticosterone continued to rise up to 16 hr then declined after 22 hr. From 28 until 48 hr corticosterone again increased significantly.
• 5.5. These results demonstrate that acute stress in female alligators causes significant suppression of plasma estradiol and a biphasic pattern of corticosterone secretion.

     

The taurine content of avian erythrocytes and its role in osmoregulation

• 1.1. The taurine content of erythrocytes from 15 avian species contained levels of taurine in the range of 20–70 mmol/kg of hemoglobin, about 100-fold that of mammalian red blood cells.
• 2.2. This high taurine content did not appear to be related to the nucleation of these cells as nucleated amphibian erythrocytes and human reticulocytes contained low levels.
• 3.3. The erythrocytes lacked cysteine sulfinic acid decarboxylase, a key enzyme in the synthesis of taurine from cysteine, indicating a probable lack of synthetic capabilities.
• 4.4. The cells were able to accumulate labeled taurine against a concentration gradient. This uptake was inhibited by β-alanine and was Na⁺-dependent.
• 5.5. When incubated in hypotonic medium, the cell volume of pigeon erythrocytes rapidly increased and was followed by a much slower return to normal size. The cell volume reduction was accompanied by a slow efflux of taurine into the medium.
• 6.6. These data suggest that taurine plays a role in cell volume maintenance and osmotic regulation in avian erythrocytes.

     

Increased muscle glucose uptake in response to chronic glyburide treatment is not related to changes in glucose transporter (GLUT4) protein

• 1.1. The mechanism of action of glyburide (a sulfonylurea) on muscle has been investigated by measuring glucose uptake and glucose transporter (GLUT4) protein levels after chronic glyburide treatment.
• 2.2. A dietary induced insulin resistant rat model (4 wk of high-fat, high-sucrose feeding) was given glyburide (2mg/kg/day) for 10 days and glucose uptake was measured in a perfused hindquarter preparation.
• 3.3. Protein levels of the GLUT4 glucose transporter were determined by Western analysis.
• 4.4. After 7 days of treatment, rats fed glyburide had lower blood glucose concentrations 2 hr (72 ± 5 vs 103 ± 12 mg/dl) and 24 hr (97 ± 7 vs 123 ± 7 mg/dl) after glyburide administration with no difference in serum insulin levels compared to vehicle treated animals.
• 5.5. Glucose uptake was approx doubled in basal state (0 insulin) in response to glyburide (2.8 + 0.4 vs 1.7 ± 0.2μ mol/g per hr).
• 6.6. Maximal insulin (100 nM) stimulated glucose uptake tended to be higher in the glyburide treated group, but did not reach statistical significance (8.0 ± 0.7 vs 7.0 ± 0.6 μmol/g per hr).
• 7.7. Western analysis revealed no significant effect of glyburide on the GLUT4 protein level in skeletal muscle.
• 8.8. These results suggest that glyburide alters glucose uptake through some mechanism other than alterations in the level of the GLUT4 glucose transporter protein.

     

Purification of the flavin-containing monooxygenase from mouse and pig liver microsomes

• 1.1. The microsomal flavin-containing monooxygenase has been purified from mouse and pig liver utilizing Cibacron-Blue Sepharose, Procion-Red agarose, and 2'5'-ADP Sepharose.
• 2.2. The enzymes had a final specific activity of 1200 and 954 nmol/min/mg protein from mouse and pig liver respectively.
• 3.3. The enzyme from both mouse and pig liver displayed typical flavoprotein spectra and appeared homogeneous by denaturing polyacrylamide gel electrophoresis.

     

Characteristics of Crassostrea virginica crystalline style chitin digestion

• 1.1. Fundamental chitin digestion characteristics of Crassostrea virginica crystalline style were investigated.
• 2.2. Optimum temperature and pH were 34°C and 4.8. respectively.
• 3.3. The colloidal regenerated chitin (0.56mol/0.5 ml: GlcNAc equivalents) was saturating under all enzyme levels encountered.
• 4.4. There was no evidence of end product inhibition, even after 100 hr incubation.
• 5.5. Calculated K_m for the chitinase complex was 1.19mM when determined using a 30 min assay, but was only 0.70 mM when determined using a 4.6 hr assay.
• 6.6. Both K_m values are lower than reported for similar assays in other molluscs and for most bacteria.
• 7.7. Effect of substrate preparation on the kinetics are discussed.
• 8.8. Eight peaks of chitinase activity were resolved by DEAE-Fractogel ion exchange chromatography.

     

The role of citrate derived from glucose in the acetylcholine synthesis in rat brain synaptosomes

• 1.1. Synaptosomes utilizing glucose or glucose plus malate produced citrate with rates of 2.4 and 7.8 nmol/hr/mg of protein, respectively.
• 2.2. (−)Hydroxycitrate increased citrate net synthesis 4 times and inhibited acetylcholine synthesis by 40%.
• 3.3. Oxygen and glucose consumption as well as lactate and CO₂ production were not changed by this inhibitor.
• 4.4. (−)Hydroxycitrate inhibited utilization of exogenous citrate in synaptosomes by 50%.

     

Renal function in migrating adult atlantic salmon,Salmo salar L.

• 1.1. Renal function in migrating adult Atlantic salmon was studied in sea-water (SW) and following abrupt transfer to fresh water (FW).
• 2.2. Urine flow rate of SW-adapted fish, 0.72 ml/kg/hr, increased 6.3-fold to 4.55 ml/kg/hr after 2–3 days in FW, later decreasing to around 1 ml/kg/hr.
• 3.3. Changes in glomerular filtration rate and ion filtration rates largely paralleled changes in urine flow. In SW-adapted salmon about 4% of excreted magnesium is filtered. Tubular magnesium secretion declined within 1 day of FW transfer.
• 4.4. During the period of maximum diuresis, urinary sodium loss is 77% of the branchial sodium uptake rate. This falls to less than 20% in FW-adapted fish.

     

The effect of feed restriction on certain haematological indices,enzymes and metabolites in Nubian goats

• 1.1. Some effects of restricting feed intake for 96 or 168 hr were determined in male Nubian goats.
• 2.2. Goats restricted for 96 hr lost 11.6% of their body weight, and goats restricted for 168 hr lost 19.8%.
• 3.3. Feed restriction for up to 168 hr did not produce significant effects on the heart rate, respiratory rate or rectal temperature.
• 4.4. Haemoglobin concentration, packed cell volume and erythrocyte number were all decreased by feed restriction. There was also a tendency towards eosinopenia and lymphopenia.
• 5.5. Feed restriction for 96 or 168 hr raised the plasma activity of aspartate transaminase, and did not affect significantly cholinesterase activity. Plasma amine oxidase activity was significantly reduced in goats restricted for 168 hr.
• 6.6. Feed restriction produced significant increases in the blood or plasma concentrations of lactate. pyruvate, non-esterified fatty acids, cholesterol, ketone bodies and bilirubin.
• 7.7. Significant decreases were found in the concentrations of total protein and calcium.
• 8.8. No significant changes were observed in the plasma concentrations of glucose, sodium or potassium.

     

Circadian rhythms of determined blood chemistry values in buzzards and eagle owls

• 1.1. Fifteen values were determined in blood samples from six buzzards (Buteo buteo) and six eagle owls (Bubo bubo) over the 24 hr of the day.
• 2.2. Glucose, urea, uric acid, triglyceride and calcium values showed diurnal rhythms in both species. Their respective patterns of diurnal variation were compared.
• 3.3. Phosphorus, cholesterol and cholinesterase levels underwent circadian rhythms only in the buzzards. Albumin/globulin and amylase exhibited diurnal variations exclusively in the eagle owls.
• 4.4. Glutamatic oxaloacetic transaminase, albumin, globulin, total protein and creatinine concentrations did not show diurnal rhythms in either of the species.
• 5.5. Blood values of the different parameters were studied on the basis of the ranges described in birds.

     

The effect of hypoxia on carbohydrate metabolism in flounder (Platichthys flesus L.)—I. Utilization of glycogen and accumulation of glycolytic end products in various tissues

• 1.1. Resting oxygen consumption at 10°C did not change from normoxia (150 mm Hg) down to an oxygen tension of 55 mm Hg for the flounder, Platichtys flesus.
• 2.2. Flounders exposed to hypoxia showed increased levels of blood glucose and lactate, dependent on the degree of hypoxia.
• 3.3. Due to hypoxia glycogen was depleted in the liver and swimming muscle but in the heart there was no significant change.
• 4.4. Liver glucose increased after 7 hr of hypoxia. Heart and muscle glucose did not change but the absolute glucose concentration in the heart was five times higher than in the muscle.
• 5.5. There is a transient accumulation of lactate in heart, liver and kidney after 7 hr of hypoxia while lactate accumulation in the swimming muscle is significant only after 21 hr of hypoxia.
• 6.6. Succinate only accumulated in the liver while alanine accumulated in muscle, heart and liver.

     

Carbohydrate metabolism in several tissues of rainbow trout,Oncorhynchus mykiss,is modified during ovarian recrudescence

• 1.1. Several pathways of carbohydrate metabolism were evaluated in three different tissues—liver, gonad and kidney—of a hatchery-reared population of rainbow trout (Oncorhynchus mykiss) which characterised two different stages of their gonadal maturation, i.e. previtellogenesis and established exogenous vitellogenesis.
• 2.2. A fall in liver glycogen levels was observed during exogenous vitellogenesis. A decrease in activity of the enzymes involved in glycolysis and in the pentose phosphate shunt was also observed, suggesting that at the end of exogenous vitellogenesis the necessity of energy and reducing power has decreased compared to the situation at the onset of this period.
• 3.3. The main changes observed in gonad during vitellogenesis were the decreased activity of glycolysis and the pentose phosphate shunt as well as increased glycogen levels. The stored glycogen should be used later in association with the embryo development.
• 4.4. No major changes were observed in kidney metabolism throughout the vitellogenic process.
• 5.5. Exogenous vitellogenesis in rainbow trout is mainly associated with increased glycogen levels in the gonad and decreased metabolic activity in the liver.

     

Energy metabolism of human erythrocytes in psoriasis

• 1.1. Adenine nucleotide concentrations and metabolism in red blood cells (RBC)2 and RBC ghosts from psoriatic patients and healthy subjects were compared.
• 2.2. The ATP and total adenine nucleotide levels and the adenylate energy charge (EC) were elevated in the blood from psoriatic patients.
• 3.3. The rate of glycolytic production of ATP by intact RBC was unchanged, but the Na⁺, K⁺-ATPase activity of RBC ghosts was decreased significantly in psoriasis.
• 4.4. Results suggest that the defect in adenine nucleotide metabolism is a systemic manifestation of psoriasis, and that the quantification of adenine nucleotides in RBC and in whole blood samples may be of pathophysiological value in psoriatic lesion.

     

Acute hepatic porphyria syndrome with porphobilinogen synthase defect

On the basis of metabolite and enzyme studies a new type of acute hepatic porphyria with porphobilinogen synthase defect and repeated intermittent acute manifestations, abdominal colics, tachycardia and hypertension, and a persistent neurological syndrome was found in two young male patients. The main characteristic features are the following:

• 1.1. High urinary δ-aminolevulinic acid excretion( ⪢ 1 mmol/24hr), slight increase of porphobilinogen (up to 25 μmol/24 hr) and high increase of porphyrins (up to 22 μmol/24 hr) with coproporphyrin dominance.
• 2.2. Normal fecal and liver porphyrins.
• 3.3. Slight increase of erythrocyte protoporphyrin.
• 4.4. Decrease of porphobilinogen synthase activity in erythrocytes in both cases below 1% of healthy and not lead-exposed persons; normal activities of uroporphyrinogen synthase and decarboxylase in erythrocytes.
• 5.5. Low-normal lead concentrations in blood and low-normal lead excretion in urine in both cases; normal lead content in bone.
• 6.6. Normal plasma and urinary amino acids.
• 7.7. Irrelevant hepatological (liver biopsy), general clinical chemical and hematological findings.
• 8.8. Diminished activity of porphobilinogen synthase in nearly all family members of both patients. From these investigations it can be concluded that there is no exogeneous, “toxic” cause of this porphyria. Porphobilinogen synthase in lead poisoning is not diminished to such an extent as demonstrated here; in contrast to lead intoxication, porphobilinogen synthase activity cannot be activated or reactivated by thiols. All clinical and pathobiochemical data point at a new enzymatic type of endogeneous acute hepatic porphyria with intermittent acute manifestations, clinically analogous to so-called acute intermittent porphyria. Porphyrin precursors and porphyrin excretion both reflects the enzymatic defect and the regulatory consequences starting with the induction of δ-aminolevulinic acid synthase.

     

A novel inhibitor of glycogen phosphorylase from crayfish hepatopancreas

• 1.1. A novel glycogen phosphorylase inhibitor was partially purified from crayfish hepatopancreas.
• 2.2. The inhibitor was found only in two species of crayfish examined, and not in lobster, fresh and salt water clams, mussels or cockroaches.
• 3.3. The inhibitor is a small protein (M_r = 23,000) which did not show proteolytic activity.
• 4.4. Preliminary kinetic analysis of the inhibitory mechanism indicated that it bound to both glycogen and the glycogen phosphorylase protein.
• 5.5. Inhibitor binding to glycogen resulted in a competitive inhibition pattern with respect to glycogen phosphorylase (inhibition constant of ca 10 μg/ml).
• 6.6. The inhibitor also bound glycogen phosphorylase directly with a binding coefficient of 100 μg/ml resulting in a partially non-competitive inhibition pattern with respect to phosphate.