Insulin administration induces capillary growth in brown adipose tissue of heat-exposed rats

• 1.1. An apparent effect of insulin administration on enlargement of interscapular brown adipose tissue (BAT) was found in heat-exposed rats, but not in warm-adapted or cold-acclimated rats.
• 2.2. BAT extracts from the heat-acclimated/insulin-treated (HI) rats notably increased the capillary growth in an in vitro angiogenesis model in which microvascular fragments and myofibroblastic (Mf) cells isolated from lipid tissues were grown in co-culture, although a direct effect of insulin was not high.
• 3.3. BAT extracts from the HI rats stimulated the production of endothelial cell growth factor and collagen by Mf cells.
• 4.4. It is probable that an increased angiogenic activity contributes to the capillary growth and tissue growth in BAT of HI rats.

     

Skin blood flow in the Wistar-Kyoto rat and the spontaneously hypertensive rat

• 1.1. Using laser Doppler techniques in man, we have previously demonstrated differences in skin blood flow properties at sites with primarily nutritive (NUTR) perfusion, such as the elbow or knee, as compared to sites such as the finger pulp, with predominantly arteriovenous anastomotic (AVA) perfusion.
• 2.2. Basal and heat stimulated flow is greater at AVA sites. In man, blood pressure changes are reflected primarily by changes at AVA rather than NUTR sites.
• 3.3. These blood pressure induced changes affect the red blood cell velocity (VEL) component at AVA sites more than microvascular volume (VOL).
• 4.4. Given these findings in man, we decided to compare skin blood flow properties in a suitable animal model.
• 5.5. We chose the Wistar-Kyoto (WKY) and Spontaneously Hypertensive Rat (SHR) strains, in view of the marked difference in systemic blood pressure in these two related strains.
• 6.6. Skin blood flow varied considerably at different skin sites in the rats. Skin sites with hair covering, on the back and at the base of the tail, showed low basal and heat stimulated blood flow.
• 7.7. In contrast, the plantar surface of the paw behaved similarly to the finger or toe pulps in man, with 3–4-fold higher basal flow than the hair covered areas and a 7–8-fold rise with local heating to 44°C.
• 8.8. Furthermore, there was a 25% greater blood flow at the plantar paw surface in the SHR rats as compared to the WKY rats, corresponding to the 25% higher systemic blood pressure in these animals.
• 9.9. The heat induced increase in flow at the plantar surface of the paw was primarily a result of a marked increase in VEL rather than VOL.
• 10.10. The higher flow at this site in SHR as compared to WKY rats was likewise ascribable to an increase in VEL, VOL being equivalent in the two strains.

     

Measurement of blood flow in the skin and oral mucosa of the rhesus monkey (Macaca mulatta) using laser doppler flowmetry

• 1.1. It is generally assumed that oral blood flow is higher than that of skin, and invasive methods to measure blood flow support this view.
• 2.2. However, it was not known whether this finding would be confirmed by laser Doppler flowmetry, which is a noninvasive method to measure blood flow.
• 3.3. The purpose of this study was to compare blood flow in oral and skin regions of the rhesus monkey using laser Doppler flowmetry.
• 4.4. The results demonstrated that blood flow was significantly higher in oral regions as compared to facial skin (P < 0.05).
• 5.5. This finding is most likely related to the more abundant capillary supply in oral mucosa as compared to skin.

     

Effects of the nonsteroidal anti-inflammatory drug piroxicam on energy metabolism in the perfused rat liver

• 1.1. The actions of piroxicam, a nonsteroidal and noncarboxylic anti-inflammatory drug, on the metabolism of the isolated perfused rat liver were investigated. The main purpose was to verify if piroxicam is also active on glycogenolysis and energy metabolism, as demonstrated for several carboxylic nonsteroidal anti-inflammatories.
• 2.2. Piroxicam increased oxygen consumption in livers from both fed and fasted rats.
• 3.3. Piroxicam increased glucose release and glycolysis from endogenous glycogen (glycogenolysis).
• 4.4. Gluconeogenesis from lactate plus pyruvate was inhibited.
• 5.5. The action of piroxicam on oxygen consumption was blocked by antimycin A, but not by atractyloside.
• 6.6. The action of piroxicam in the perfused rat liver metabolism seems to be a consequence of its action on mitochondria.
• 7.7. It can be concluded that inhibition of energy metabolism and stimulation of glycogenolysis are not specific properties of carboxylic nonsteroidal anti-inflammatory drugs.

     

Sex differences in vascular reactivity to adrenergic agents in the rat

• 1.1.The study was designed to determine if there are sex-dependent differences in vascular reactivity to adrenergic agents.
• 2.2.Vascular reactivity of both aortic and tail artery smooth muscle from male and female rats to various vasoactive agents was assessed. 3.li]The vascular response of aortic smooth muscle to both phenylephrine and isoproterenol were significantly greater in male rats as compared to females.
• 3.4.There were apparent sex differences in responsiveness to the KCl-induced, non-receptor mediated contraction of aortic smooth muscle in that the sensitivity to KCl was enhanced in male rats.
• 4.5.No sex differences were observed in tail artery preparations.
• 5.6.Phentolamine reduced the maximal tension induced by KCl in the tail artery but not aortic artery preparations. This effect was not sex dependent.
• 6.7.No differences in the vascular smooth muscle responsiveness to acetylcholine or sodium nitrate was observed between groups or within different vascular beds.
• 7.8.The increased sensitivity of males to adrenergic challenge could explain in part some of the existing sex differences in cardiovascular disease and hypertension.

     

The effect of d2o on glycolysis by rat hepatocytes

• 1.1. The effect of incorporating D₂O into the incubation medium on glycolysis and gluconeogenesis by hepatocytes from fasted rats was examined.
• 2.2. The substitution by heavy water, D₂O, at concentrations from 10 to 40%, stimulated glucose uptake, lactate production and CO₂ yields from glucose. At 10 mM glucose, 40% D₂O doubled glucose uptake, increased CO₂ production by 40%, and increased lactate production by 350%.
• 3.3. The stimulation of lactate production decreased at higher glucose concentrations, but was still substantial even at 80 mM glucose.
• 4.4. There was no effect on CO₂ production above glucose concentrations of 30 mM.
• 5.5. Ten percent D₂O showed little inhibition of lactate uptake, its oxidation and gluconeogenesis. At 40% D₂O the inhibition ranged from 10 to 20%.
• 6.6. No effect of D₂O on the rate of glucokinase or glucose-6-phosphatase was observed.
• 7.7. The concentration of fructose, 2,6-P was not affected by D₂O

     

Effect of methyl methacrylate on mitochondrial function and structure

• 1.1. Treatment of isolated rat liver mitochondria with methyl methacrylate (MM) produced membrane disruption as evidenced by the release of citrate synthase, and changes in the ultrastructure of mitochondria.
• 2.2. At concentration 0.1%, MM uncoupled oxidative phosphorylation as evidenced by stimulation of state 4 respiration supported either by pyruvate plus malate or succinate (+rotenone) and ATP-ase activity in intact mitochondria.
• 3.3. At concentration 1% MM stimulated ATP-ase activity in intact mitochondria and succinate (+rotenone) oxidation at state 4 and was without effect on this substrate oxidation at state 3.
• 4.4. MM inhibited pyruvate plus malate oxidation either at state 3 or in the presence of uncoupling agents.
• 5.5. MM inhibited the NADH oxidase of electron transport particles at a concentration which failed to inhibit either succinic oxidase or the NADH-ferricyanide reductase activity.
• 6.6. The data presented suggest that in the isolated mitochondria MM inhibits NADH oxidation in the vicinity of the rotenone sensitive site of complex I.
• 7.7. The general conclusion is that MM may block an electron transport and to uncouple oxidative phosphorylation in rat liver mitochondria. The overall in vitro effect would be to prevent ATP synthesis which could result in cell death under in vivo conditions.

     

Locality-dependent concentrations of bromophenols in Lanice conchilega (Polychaeta: Terebellidae)

• 1.1. At 37 stations in the English Channel and the southern North Sea concentrations of four brominated phenols in Lanice conchilega were determined using ECD equipped capillary gas chromatography: 2,4-dibromophenol, 2,6-dibromo-4-methylphenol, 2,4,6-tribromophenol, 3,5-dibromo-4-hydroxybenzaldehyde.
• 2.2. Concentrations in worms of the southern North Sea were generally below 1 μg/g wet wt.
• 3.3. Levels were slightly raised in worms of sheltered shores, those of 3,5-dibromo-4-hydroxybenzaldehyde were increased in subtidal populations.
• 4.4. Concentrations in L. conchilega of Brittany were considerably higher than those in worms of the Frisian Islands, North Sea; they deviated from the latter by factors of 3, 16, 4, 4, respectively.
• 5.5. The reasons for the conspicuous differences are hitherto unknown; three explanations are suggested.

     

Reduced muscle protein breakdown in septic rats following treatment with interleukin-1 receptor antagonist

• 1.1. The role ofinterleukin-1 (IL-1) in sepsis-induced muscle proteolysis was assessed by treating septic rats with recombinant IL-1 receptor antagonist (rIL-Ira).
• 2.2. In initial experiments, we tested the effectiveness of IL-Ira in preventing muscle proteolysis induced by administration of IL-1.
• 3.3. When normal rats were treated with rIL-α (three intraperitoneal doses of 100 μ g/kg body weight each over 16 hr), total and myofibrillar muscle protein breakdown rates, measured as release oftyrosine and 3-methylhistidine, respectively, by incubated extensor digitorum longus muscles, were significantly increased.
• 4.4. This metabolic response to IL-α was completely abolished by rIL-Ira, administered as three intraperitoneal doses of 3 mg/kg body weight each over 16hr.
• 5.5. In subsequent experiments, sepsis was induced in rats by cecal ligation and puncture (CLP); non-septic rats were sham-operated.
• 6.6. Treatment of septic rats over 16hr with a total dose of 25mg/kg body weight of rIL-Ira reduced, but did not normalize, the increased muscle protein breakdown rates seen during sepsis.
• 7.7. When the dose of rIL-Ira was more than doubled and given as a constant infusion at a rate of 4.2 mg/kg body weight/hr for 16 hr, the increased rate of muscle proteolysis in septic rats was normalized.
• 8.8. The present study offers the first direct evidence that IL-1 is involved in the regulation of muscle proteolysis during sepsis.

     

Effects of high dietary methionine on activities of selected enzymes in the liver of kittens (felis domesticus)

• 1.1. Growing male kittens were fed an 18% casein diet supplemented with 2, 3, or 4% l-methionine (MET) for 6 weeks.
• 2.2. Free MET concentration in liver increased 30-fold and cystathionine two- to three-fold; the activity of adenosyl-MET transferase and cystathionase also increased but remained lower than previously found in rats.
• 3.3. Taurine concentration in liver decreased in cats fed excess MET and appeared to depend on taurine intake.
• 4.4. Alanine aminotransferase activity was high in all groups while serine dehydratase activity was very low.
• 5.5. Pyruvate kinase and malic enzyme activities which are normally low in cat liver increased after excess MET. Also, glucose 6-phosphate and 6-phosphogluconate dehydrogenases increased.
• 6.6. Cat liver metabolism showed limited adaptation to an excess dietary intake of methionine compared to that found in rats.

     

Effect of lindane on phosphatidylinositol synthesis by cerebral cortex after acute and subchronic treatment

• 1.1. The incorporation of myo-[2-³H]inositol into phosphatidylinositols was unmodified in brain cortex miniprisms from convulsant rats.
• 2.2. However, the incorporation had increased by 300–400% in non convulsant rats which had received the same amount of lindane at a lower concentration.
• 3.3. This result suggests that phosphatidylinositols are implicated in the convulsion syndrome.
• 4.4. Experiments with lindane added in vitro were performed with both subchronically lindane intoxicated and untreated rats.
• 5.5. The results show an interesting lack of parallelism.
• 6.6. This might indicate the development of some resistance to the effects of lindane, possibly as the result of complex compensatory changes in inositol lipid biosynthesis.

     

Properties of the transition from short-term to long-term depression in neurons of Helix pomatia

• 1.1. Synaptic short-term depression could be transferred into long term depression by repetition of series of stimuli.
• 2.2. The transition from short-term depression to long-term depression was blocked by puromycin.
• 3.3. The majority of the transition took place during resting periods between stimulus series.
• 4.4. The initiation of the transition process was 83% completed after 5 min of stimulation.
• 5.5. Short- and long-term depression were quantitatively separated into their two serial sites of origin: afferent axons and synaptic terminals.
• 6.6. Long sequences evoked periods with increased and variable EPSPs not conforming to depression.

     

Differential post-receptor responses of adenylate cyclase in white and brown adipose tissue membranes of rats fed high-energy diets

• 1.1. Adenylate cyclase activity was determined in membranes of white and brown adipose tissue (WAT and BAT, respectively) from rats fed a high-energy diet (EXP group) vs those fed a nutritionally balanced one (CON group).
• 2.2. The isoproterenol- and guanine nucleotide-induced adenylate cyclase activity in WAT membranes of EXP rats was lower than that in CON rats.
• 3.3. Relative adenylate cyclase activity in like treated BAT membranes was higher in EXP than in CON rats.
• 4.4. It is concluded that feeding high-energy diets to rats induces similar post-receptor modifications of adenylate cyclase as found in genetic obese rodents.

     

D-d-lactoylglutathione accumulation in activated human platelets

• 1.1. S-d-Lactoylglutathione accumulates in human platelets activated by agonists. Among the tested inducers thrombin is the most active.
• 2.2. The effect is dose and time-dependent. S-d-Lactoylglutathione, corresponding to depleted pool of reduced glutathione, can also be detected in platelets incubated with exogenous methylglyoxal.
• 3.3. A further significant increase was observed in platelets stimulated with trombin in the presence of methylglyoxal.
• 4.4. No change in glyoxalase activities upon platelet stimulation with thrombin was shown.

     

FlashPack: Fast and Simple Preparation of Ultrahigh-performance Capillary Columns for LC-MS*

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Highlights

• •Fast and simple capillary column packing protocol.
• •Low-pressure packing at <100 bars from ultrahigh sorbent suspension concentration.
• •Sorbent particle aggregation leading to blocking of the column entrance is avoided.
• •Effective for long capillary UHPLC column packing with a wide range of sorbents.

     

Identification of neurons involved in the earthworm Amynthas hawayanus reflex activity

• 1.1. The medial (MGF), lateral (LGF) and motor (RMS-2) giant neurons were confirmed as neural components in the earthworm Amynthas hawayanus polysynaptic reflex circuit by simultaneous potential recording and dye injection.
• 2.2. The reflex was initiated from the mechanoreceptors when evoked by mechanical stimulation but electrical stimulation also evoked an antidromic response in the motoneuron.
• 3.3. The primary reflex response propagates decrementally along both giant axons but directly evoked action potentials conduct in an all-or-none fashion.
• 4.4. The secondary reflex response continues to propagate after the primary response disappears.
• 5.5. A rhythmically discharging neuron of uncertain function was also identified.

     

Fiber type composition and enzyme activities of muscles in two models of obese rats

• 1.1. In oxidative (soleus) and glycolytic (extensor digitorum longus) muscles of obese Zucker rats, a significant decrease in the percentage of relative area occupied by glycolytic fibers was observed.
• 2.2. The activity of citrate synthase and β-hydroxy-acyl-CoA-dehydrogenase was significantly higher in muscles of obese than of lean Zucker rats.
• 3.3. In rats, 6 weeks after lesion of the ventromedial hypothalamus, no changes were observed.
• 4.4. This indicates that neither the proportion of oxidative fibers, nor the oxidative capacities are decreased in skeletal muscles of obese rats suggesting that insulin resistance cannot be ascribed to a higher glycolytic-oxidative fiber ratio.

     

Modulation of mitochondrial ATPase sensitivity to inhibitors and stimulators by diet-induced changes in membrane lipid

• 1.1. Weanling rats were fed diets differing in fatty acid composition to determine if changes induced in cardiac mitochondrial membrane structural components alter the sensitivity of mitochondrial ATPase to inhibition by oligomycin and stimulation by 2,4-dinitrophenol.
• 2.2. Mitochondrial ATPase assayed in situ within the mitochondrial membrane isolated from animals fed diets higher in fatty acids of longer chain length, exhibited greater oligomycin sensitivity and lower 2,4-dinitrophenol-induced stimulation.
• 3.3. Concomitant diet-induced changes occur in the fatty acid, composition of phosphatidylcholine, phosphatidylethanolamine and cardiolipin, increasing overall length of fatty-acyl tails in the membrane phospholipids.
• 4.4. Diet fat mediated alterations in oligomycin sensitivity of mitochondrial ATPase and membrane fatty acid chain length suggest that _vivo changes in thickness of the lipid bilayer may alter mitochindrial ATPase functions.
• 5.5. The present study extends the concept that dietary fat affects mitochondrial membrane structure and function by demonstrating that the membrane-dependent sensitivity of mitochondrial ATPase to inhibitors and stimulators may be modulated by dietary fat.

     

Role of glutamate dehydrogenase reaction in the control of citrate pool in yeast

• 1.1. Role of NADP-glutamate dehydrogenase in the depletion of citrate was analyzed using permeabilized yeast cells.
• 2.2. Citrate was converted to 2-oxoglutarate, which was then metabolized to glutamate by NADP-glutamate dehydrogenase in the presence of ammonium ion.
• 3.3. Formation of 2-oxoglutarate plus glutamate was in good agreement with the concentration of citrate decreased. Glutamate formation can be a good indicator of the depletion of citrate, because 70% of the citrate decreased was converted to glutamate.
• 4.4. Glycolytic activity was closely correlated with the decrease in citrate under the in situ conditions.
• 5.5. NADP-glutamate dehydrogenase increased in anaerobically grown yeast cells.
• 6.6. An effective depletion of citrate by increased synthesis of NADP-glutamate dehydrogenase can explain the lowered mechanism of citrate causing glycolytic stimulation under the anaerobic growth conditions of yeast.

     

Anaerobic threshold in rats

• 1.1. The aim of this study was to find out whether the anaerobic threshold (AT) can be estimated in rats running at increasing speed and if so what is the reproducibility of the measurements.
• 2.2. Lactate (LA) concentrations in blood taken from 11 rats were determined during a discontinued, multistage treadmill exercise test repeated four times in each animal.
• 3.3. It was found that blood LA changes vs speed have an exponential pattern with a distinct, rapid rise at the speed above 25 m/min which corresponds to blood LA of approx. 4 mmol/1.
• 4.4. The variation coefficient of the speed at which AT occurred in individual animals ranged between 10 and 20%.
• 5.5. These results offer a potential application of AT determination in the animal studies concerning mechanisms controlling exercise metabolism.