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.

     

Alterations in proteases,protease inhibitors and ecdysone levels: a profile of stress in insects

• 1.1. A comparison of proteolytic and protease inhibitory activity, and ecdysteroid levels in body fluids was made between normal larvae of the flesh fly, Sarcophaga bullata, and those that had been water-stressed for two days.
• 2.2. The course of proteolytic activity in water stressed flies decreases 6 hr after beginning the experiment and remains low in comparison with control flies.
• 3.3. The course of protease inhibitors exhibits a mirror image pattern to proteases.
• 4.4. Ecdysteroid pattern shows two peaks in control animals: minor at 24 hr and major at pupariation, in experimental animals: at 1 hr, at 6 hr and at white pupal stage.

     

Laboratory studies on the oxygen consumption of the marine teleost,Lichia amia (Linnaeus, 1758)

• 1.1. The oxygen consumption of the marine teleost, Lichia amia was investigated under controlled laboratory conditions.
• 2.2. The routine oxygen consumption showed a strong circadian rhythm with the fish being mainly active during the light period.
• 3.3. The specific mass exponent (dimension: μg O₂/g/hr) is temperature independent and ranges from 0.27–0.29.
• 4.4. Starving the fish results in a mean decrease in active, routine and standard oxygen consumption of 21%, 24% and 20%, respectively.
• 5.5. Feecling led to an increase in the oxygen consumption of the teleosts, with the mean metabolic rate over the 24 hr that followed, being 58% and 50% higher for fish that had been starved for 162hr and 40 hr, respectively.
• 6.6. Apparent SDA showed some variation and ranged from 6.0 to 35.5%.
• 7.7. The results obtained are generally in agreement with those recorded for other teleosts.

     

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.

     

Heat shock proteins from the lungless salamanders Eurycea bislineata and Desmognathus ochrophaeus

• 1.1. Exposing adult salamanders, Eurycea bislineata and Desmognathus ochrophaeus, to heat shocks of 1 hr at 2 or 5°C below Critical Thermhal Maximum (CTM) resulted in the induction of two heat shock proteins (hsps) of approx. M_r 70,000 and 30,000.
• 2.2. Induction patterns in response to similar heat shocks generally differed between the two species.
• 3.3. The milder heat shocks (5°C below CTM) caused different induction patterns than those from the more severe heat shocks, on a tissue-dependent basis. These results indicate that induction of the two hsps is probably independent.

     

The effects of temperature and moisture on survival capacity,cuticular permeability,hemolymph osmoregulation and metabolism in the scorpion,Centruroides hentzi (banks) (scorpiones,buthidae)

• 1.1. The temperature and water relations of Centruroides hentzi females were investigated. At 12 and 72% relative humidity (RH), the lower and upper Lt₅₀ were -4.5 and 43.7°C, and -4.7 and 45.1°C, respectively. When exposed to high temperature stress, survivorship was significantly greater under mesic conditions.
• 2.2. Cuticular water loss was higher under xeric conditions (12% RH), ranging from 0.061 mg/cm²/hr at 30°C to 0.211 at 41°C.
• 3.3. Exposure to dry air (0–5% RH) resulted in a significant increase in hemolymph osmolality: from 441 to 688 mOsm over a 5 day period.
• 4.4. Mean oxygen consumption rates increased from 161.7 mm³/g/hr at 34°C to 541.6 at 44°C. ATPase activity was significantly higher in animals acclimated and tested at 35°C.

     

Changes in free amino acid composition in haemolymph of larvae of the wax moth,Galleria mellon ella L., during cold acclimation

• 1.1. Free amino acids were analysed in the haemolymph of Galleria mellonella larvae by HPLC chromatography with o-phthaldialdehyde (OPA)-l-thio-β-d-glucose as derivatization agent.
• 2.2. Fourteen primary amino acids were detected among which glutamine, alanine, γ-aminobutyric acid (GABA) and glycine predominated and constituted 67.7% of the amino acids found.
• 3.3. The concentration of GABA increased significantly with the age of larvae entering the wandering phase and reached a maximum during metamorphosis.
• 4.4. Analysis of cold-acclimated larvae revealed a net increase of free primary amino acids from 96 to 151.8 μmol/ml during consecutive acclimation to 0°C within 20 days and to 205.4μmol/ml during cold shock injury at 0°C (3 hr).
• 5.5. The bulk of this increase was accounted for by alanine, glycine, phenylalanine and lysine.

     

Effect of inorganic phosphate on synthesis of 5-phosphoribosyl-1-pyrophosphate in cultured plant cells

• 1.1. Inorganic phosphate (Pi) was absorbed rapidly by suspension-cultured cells of Catharanthus roseus which had previously been cultured in Pi-free Murashige Skoog medium.
• 2.2. The intracellular levels of ATP, ADP and 5-phosphoribosyl-l-pyrophosphate (PRPP) increased markedly during the 24 hr which followed the addition of Pi (1.25mM).
• 3.3. Availability of PRPP in vivo, estimated by the measurement of nucleotide synthesis from [8-¹⁴C]adenine, was also increased by addition of Pi.
• 4.4. Only a 20% increase in the maximum catalytic activity of PRPP synthetase was observed in extracts of cells, prepared 24 hr after addition of Pi.
• 5.5. In contrast to results for mammalian PRPP synthetase, the activity of PRPP synthetase, partially purified from Catharanthus roseus, was inhibited by concentration of Pi greater than 5mM.
• 6.6. The mechanisms involved in the increased availability of PRPP and the synthesis of adenine nucleotides in the plant cells cultured in Pi-containing medium are discussed.

     

Changes in high-energy phosphate metabolism in the water scorpion,Ranatra chinensis,under cold water-warm water stress

• 1.1. To evaluate changes in high-energy phosphate metabolism in the water scorpion (Ranatra chinensis) under restraint and cold water-warm water stresses, in vivo [³¹P]NMR spectra were obtained.
• 2.2. Under restraint stress, arginine phosphate (Arg-P) decreased by 10% after 1 hr and remained at that level thereafter, while β-ATP showed negligible changes over 6 hr.
• 3.3. As the water temperature gradually increased or decreased, the relative concentration of Arg-P decreased due to enzyme regulation.
• 4.4. Repeated cold water-warm water stress, which consisted of repeated 15 min exposures to cold water (5°C) followed by 15 min exposures to warm water (30°C) caused distinct decreases in Arg-P and β-ATP concentration. These decreases were dependent on the frequency of exposure.
• 5.5. Phosphomonoesters (PME) increased not only with restraint stress but also with cold water-warm water stress.
• 6.6. The effect of cold water-warm water stress on high-energy phosphate metabolism was greater than that of restraint stress.

     

Effect of temperature and salinity on the oxygen consumption of laboratory produced Penaeus californiensis postlarvae

• 1.1. The oxygen consumption by P. californiensis postlarvae (mean wt = 0.38 g) was determined at five different temperatures and four salinities.
• 2.2. The O₂ in each chamber was recorded at 10 min intervals for 1 hr. The time course of oxygen depletion was independent of O₂ concentration down to 1.6 mg/l.
• 3.3. Oxygen consumption increased with temperature from 0.0045 mg/g/min at 19°C, to 0.0142 mg/g/min at 35°C. The thermal coefficient (Q₁₀) indicated a very high sensitivity of the postlarvae to temperature variations at 19–23°C.
• 4.4. The results show that oxygen consumption significantly depends on temperature (P < 0.001) while salinity has only a marginal effect.

     

Light-induced transient increase of the activity of topoisomerase I in plasmodia of Physarum polycephalum

• 1.1. Activity of topoisomerase I and incorporation of [³H]uridine and [¹⁴C]thymidine were monitored during light-induced sporulation of the slime mold Physarum polycephalun.
• 2.2. A 4-fold transient increase of topoisomerase I activity but not of [³H]uridine or [¹⁴C]thymidine incorporation was observed after 42 hr of illumination with 6 hr impulses.
• 3.3. The activity of topoisomerase I did not increase in the absence of light impulses. However, ca 5-fold increase of the activity was observed in dark when 100 μ M dibutyryl-cAMP was administered 12 hr before harvesting of plasmodia.
• 4.4. Fluorodeoxyuridine and cycloheximide administered 36 hr after starting of the illumination cancelled the increase of the activity of topoisomerase I.
• 5.5. After 7 days of the illumination, when fruiting bodies appeared, the activity of topoisomerase I dropped to about 15% of the initial value.

     

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.

     

Energy metabolism and body temperature in 13 sunbird species (Nectariniidae)

• 1.1. Diurnal cycles of body temperature, T_b, and energy metabolism, M, at different ambient temperatures (T_a: +5 −+ 32°C) were tested in 13 sunbird species from various habitats and of different body masses (5.2–14.2 g) including one of the smallest passerines, Aethopyga christinae.
• 2.2. Resting M-level (night) reaches T_a-dependent mean values of 54% (+5°C) and 49% (+25°C) of activity M-levels (day). Expected level is ca 75%.
• 3.3. Resting metabolic rate of sunbirds lies within the range of theoretically expected values for birds.
• 4.4. Mean linear metabolism-weight regression of the night values follows: M = 0.102 × W^0.712 (M = energy metabolism in kJ/hr and W = body mass in g).
• 5.5. Thermal conductances, T_c, are lower (−24%) than the predicted values. This is caused by a decrease of T_b at low T_a. Mean nocturnal T_c is 3.2 J/g × hr × °C, mean day-time value is 4.3 J/g × hr × °C.
• 6.6. The zone of thermoneutrality is, in most species, within a T_a-range of 24–28°C.
• 7.7. Normal day and night levels of T_b are in the same range as reported for other birds of the same weight class. T_b decreases slightly with falling T_a (partial heterothermia). Lowest recorded T_b was 34.2°C.
• 8.8. No species tested showed any sign of torpor at night, independent of T_a, body mass or habitat origin.

     

Water relations and thermal sensitivity of several cockroach species (Dictyoptera: Blattidae and blaberidae)

• 1.1. Per cent total body water content (%TBW), cuticular permeability (CP), rate of water loss, critical thermal maxima (CTMax), and upper lethal limits (ULL) were determined for Pacific beetle, Diploptera punctata (Eschscholtz), Surinam, Pycnoscelus surinamensis (L.), and Turkestan, Blaita lateralis (Walker), cockroaches.
• 2.2. Initial body mass ranged from 153.16 to 464.96 mg, for D. punctata and P. surinamensis cockroaches, respectively. Mean %TBW was 57.8 for P. surinamensis and 67.7 for B. lateralis.
• 3.3. Mean cuticular permeability was not related to initial mass and ranged from 20.9 to 38.7 μg/cm²/hr/mmHg for D. punctata and P. surinamensis, respectively.
• 4.4. Cumulative mass loss and %TBW lost increased linearly with desiccation time.
• 5.5. CTMax ranged from 43.2°C for D. punctata to 44.3°C for P. surinamensis. There were significant, but small differences in CTMax among the three species.
• 6.6. ULL were 2.2 to approximately 4°C greater than CTMax. The greatest ULL was 48.1°C for B. lateralis and the lowest ULL was 45.0°C for D. punctata.

     

Regulatory role of δ-aminolevulic acid dehydratase in the dimorphic fungus Mucor rouxii

• 1.1. With the aim of finding a possible relationship between the known dimorphism phenomenon existing in the fungus Mucor rouxii and the biosynthesis of respiratory pigments, the activity of aminolevulic acid synthetase (ALA-S) and ALA-dehydratase (ALA-D) was studied in crude extracts and in 15,000 g supernatants of both mycelium and yeast-like cells.
• 2.2. The activity of ALA-S was unusually high (3 nmol ALA/hr/mg protein) compared with that reported for other tissues and did not vary with the fungus morphology.
• 3.3. Instead, ALA-D specific activity was found to be 16.5 nmol PBG/hr/mg protein in mycelium extracts, that is 7-fold greater than that measured in the yeast-like morphology (2.6 nmol PBG/hr/mg protein).
• 4.4. It was of importance to determine the activity levels of ALA-D along with the morphogenic transition from yeast to mycelium. It was observed that the greatest change and enhancing of specific activity occurred 2 hr before the emergence of the germ tubes and was held constant up to the complete development of mycelium.
• 5.5. Both hyphae formation and enhancement of ALA-D activity were diminished when cAMP was added to the culture shifted from the anaerobic atmosphere to air.
• 6.6. These findings and preliminary studies on the characterization of M. rouxii ALA-D indicate that this enzyme plays a regulatory role in porphyrin biosynthesis in this fungus as well as a key function in the characteristic morphogenic transition.

     

Absorption and blood clearance of labelled carotenoids ([14C]astaxanthin, [3H]Canthaxanthin and [3H]zeaxanthin) in mature female rainbow trout (Oncorhynchus mykiss)

• 1.1. Using one force-fed meal, eight mature female rainbow trout received [¹⁴C]astaxanthin ([¹⁴C]Ax) with [³H]canthaxanthin ([³H]Cx; N = 3) or with [³H]zeaxanthin ([³H]Zx; N = 5).
• 2.2. Approximately 200 μl of blood were collected via caudal puncture every 24 hr for 4 days. After 96 hr, the fish were killed and pyloric caeca (P.C.) from the duodenal intestine (D.I.) section, ileal intestine (I.I.), and posterior intestine (P.I.) were dissected out.
• 3.3. In the blood, Ax levels were higher than Cx followed by Zx levels.
• 4.4. This corresponds to their respective absorption by the trout as was confirmed by their relative concentrations in P.C., I.I. and P.I.
• 5.5. However, blood clearance was similar for all three compounds. [¹⁴C]Phoenicoxanthin ([¹⁴C]Px) was detected as a reduced metabolite of [¹⁴C]Ax in all gut sections.

     

Specific dynamic action demonstrated in the herbivorous marine periwinkles,LIttorina littorea L. and Littorina obtusata L. (Mollusca,Gastropoda)

• 1.1. Respiration rates of starved periwinkles, Liltorina littorea and Littorina obtusata, increased by 40–60% when fed their preferred algal food for 1 hr, or after having been exposed for the same period to an aqueous extract of the alga.
• 2.2. The stimulus causing the rise in metabolic rate by algal extracts is therefore thought to be chemosensory in nature, and possibly composed primarily of dissolved free amino acids.
• 3.3. The respiration rates of L. littorea responded only to the green alga Enteromorpha intestinalis while L. obtusata demonstrated an increase in respiration rates when fed the ubiquitous brown alga, Ascophyllum nodosum, and to a lesser degree to Enteromorpha.
• 4.4. These results compare very well to postprandial increases in oxygen consumption demonstrated in vertebrates and marine bivalves where the components of specific dynamic action (SDA) are thought to be chiefly biosynthetic costs of digestion and assimilation.

     

A two-peak circadian system in body temperature and activity in the domestic cat,Felis catus L.

• 1.1.|The body temperature and activity of cats exhibit two-peak patterns during the 24 hr period.
• 2.2.|The two peaks are retained when the temperature and activity are permitted to freerun.
• 3.3.|A third prominent peak appears in the actograms in cats in the main colony, induced by the presence of humans.

     

Induction kinetics of immune antibacterial proteins in pupae of Galleria mellonella and Pieris brassicae

• 1.1. Pupae of Galleria mellonella and Pieris brassicae given an injection with live, non-pathogenic Enterobacter cloacae or abiotic foreign molecules induce an acquired immunity that corresponds with the synthesis of haemolymph proteins of antibacterial activity.
• 2.2. This humoral defensive response which persists for several days, differs quantitatively between insect species and between the inducers used, although very different foreign bodies induced the same immune proteins in both lepidopteran insects.
• 3.3. A stronger and longer lasting response was consistently noticed in pupae immunized with non-pathogenic bacterium than after sterile nutrient broth injections.
• 4.4. A demonstrably elevated activity of haemolymph lysozyme and trace activity of cecropins found in pupae of Galleria treated with saline W, a salt solution physiological to moths, disappear soon after 36 hr from injection.
• 5.5. In P. brassicae, however, sterile insect Ringer can give a varying, if present at all, immune response.
• 6.6. A mechanical injury (sterile wounding of insect body) can occasionally induce a similar but much weaker response.
• 7.7. The antibacterial activity was drastically reduced in Pieris or completely depressed in most pupae of Galleria when actinomycin D or cycloheximide was given at an early time post-immunization with E. cloacae.
• 8.8. It is concluded that the de novo synthesis of ribonucleic acid and immune proteins is required for expression of antibacterial activity in pupal haemolymphs.
• 9.9. The synthesis of an immune mRNA was completed about 7 hr after the injection of the immunizing bacteria.

     

Purification of glycosylphosphatidylinositol-anchoring aminopeptidase in from the plasma membrane of larval midgut epithelial cells of the silkworm,Bombyx mori

• 1.1. Aminopeptidase N was selectively released from larval midgut of silkworm, Bombyx mori, by phosphatidylinositol-specific phospholipase C, and purified to a homogeneous state by ion exchange, gel filtration. Con A-Sepharose and 4-aminobenzyl phosphonic acid-agarose column chromatographies.
• 2.2. The purified aminopeptidase N preparation showed 190.8 U/mg of specific activity. Its molecular weight was estimated to be around 100 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
• 3.3. Purified aminopeptidase N molecule preferentially hydrolyzed Leu-, Ala- and Met-p-nitroanilide as substrates. Especially, Leu-p-nitroanilide proved to be the best substrate for aminopeptidase N from larval midgut of silkworm.
• 4.4. By treatment with phosphatidylinositol-specific phospholipase C, two other hydrolases, alkaline phosphatase and alkaline phosphodiesterase I, were also solubilized from silkworm midgut.