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Leukocyte-endothelial interactions in environmental hypoxia.

Hypoxia induced by reducing inspired PO2 (PIO2) to 70 Torr, promotes a rapid microvascular response characterized by increased leukocyte rolling and adherence to the venular endothelium, leukocyte emigration to the perivascular space and increased vascular permeability. This appears to be a generalized response since it is observed in venules of the mesentery, cremaster muscle and pial microcirculations. After three weeks of acclimatization to hypoxia (barometric pressure 380 Torr, PIO2 70 Torr), the initial microvascular response resolves and exposure to even lower PIO2 (50 Torr) fails to elicit a microvascular response. The initial response is accompanied by a reversible increase in the generation of reactive oxygen species (ROS) and is blocked by antioxidants and by interventions that increase the tissue levels of nitric oxide (NO). In contrast to ischemia/reperfusion, ROS levels increase during hypoxia and return towards pre-hypoxic values after return to normoxia. Acclimatization involves upregulation of inducible NO synthase (iNOS): inhibition of iNOS using two different antagonists results in increased leukocyte-endothelial interactions and increased ROS generation. The results suggest that hypoxia initially leads to an alteration of the ROS/NO balance which is eventually restored during the acclimatization process. This phenomenon may have relevance to the microcirculatory alterations associated with hypoxic exposure, including acute mountain sickness and high altitude pulmonary and cerebral edema.

Altitude Sickness↗

Ventilatory effects of prolonged systemic (CNS) hypoxia in awake goats.

Hypoxia isolated to the carotid body (CB) can induce time-dependent progressive hyperventilation (ventilatory acclimatization) in the absence of brain hypoxia. The studies reported in this paper were designed to determine if CNS hypoxia in the absence of CB hypoxia would affect ventilation over a 4 h period. In addition, the effect of 4 h of CNS hypoxia on the ventilatory responses to central chemoreceptor stimulation and to isolated CB stimulation were also determined. The studies were carried out in awake goats with CB blood gases controlled by an extracorporeal circuit while systemic (CNS) blood gases were determined independently by the level of inhaled gases. Systemic arterial PO2 was reduced to 40 Torr while the CB was maintained normoxic and normocapnic. Systemic arterial PCO2 was kept isocapnic. The data obtained indicate that 4 h of CNS hypoxia produced mild hyperventilation that reached a peak after 30 min of hypoxia and was sustained for the entire period of hypoxia. There was no evidence of a time-dependent progressive hyperventilation, i.e. no acclimatization. In contrast to studies in which whole body hypoxia is induced, CNS hypoxia did not result in any changes in the ventilatory responses to either central or peripheral chemoreceptor stimulation after return to normoxic conditions. These findings suggest no significant role for CNS mechanisms induced by hypoxia in ventilatory acclimatization to hypoxia in goats.

Animals↗

Effect of environmental temperature on rainbow trout (Oncorhynchus mykiss) innate immunity.

Phagocytosis, complement lytic activity and opsonization capacity of rainbow trout plasma as well as the ability of phagocytes to recognize foreign particles were studied at different temperatures. Respiratory burst (RB) activity and opsonization capacity were assessed as chemiluminescence emission from diluted whole blood of fish which were acclimatized for 57 days at temperatures between 5 and 20 degrees C. RB activity was higher at higher acclimatization and in vitro assay temperatures. The peak time of RB was significantly delayed in fish kept at lower temperatures (5-10 degrees C) as compared to fish kept at 15 or 20 degrees C temperatures. Opsonization capacity of plasma decreased in fish acclimatized at low temperatures and was also affected by in vitro assay temperature. The importance of glucan receptors in RB activity increased in fish kept at higher temperatures and was also affected by the in vitro assay temperature. The higher acclimation temperatures increased the lytic activity of both total and alternative complement pathways.

Animals↗

Aerobic biodegradation of organotin compounds in activated sludge batch reactors.

The biodegradation behavior of four organotin (OT) compounds, namely tributyltin (TBT), dibutyltin (DBT), monobutyltin (MBT) and triphenyltin (TPhT), was studied in lab-scale activated sludge batch reactors. The activated sludge was spiked with the OT compounds at a level of 100 microg l(-1) as Sn. Determination of the OT compounds by GC-FPD after ethylation in the dissolved and particulate phase revealed that 24 h after the start of the experiments, almost the total of OT compounds has been removed from the dissolved phase and is associated with the suspended solids. Calculation of mass balance in batch reactors showed that OT compounds biodegradation was performed via a sequential dealkylation process. Removals due to biodegradation were differentiated according to the parent compound. In experiments with non-acclimatized biomass, a percentage of 27.1, 8.3, 73.8 and 51.3 was still present as TBT, DBT, MBT and TPhT, respectively, at the end of the experiment (18th day). Half-lives (t1/2) of 10.2 and 5.1 days were calculated for TBT and DBT, respectively, whereas apparent t1/2 values could not be determined for MBT and TPhT (t1/2>18 days). The capacity of activated sludge to biodegrade OT compounds in the absence of supplemental substrate indicated that these compounds can be metabolized as single sources of carbon and energy in activated sludge systems. Excluding TBT, the presence of low concentrations of supplemental substrate did not affect the biodegradation potential of activated sludge. The acclimatization of biomass on OT compounds enhanced significantly biodegradation, resulting in significant decreases of half-lives of OT compounds. As a result in the presence of acclimatized biomass, half-lives of 1.4, 3.6, 9.8 and 5.0 days were calculated for TBT, DBT, MBT and TPhT, respectively.

Biodegradation, Environmental↗

Monitoring the stability of Rubisco in micropropagated grapevine (Vitis vinifera L.) by two-dimensional electrophoresis.

Plants cultured in vitro suffer from several physiological and biochemical impairments due to the artificial conditions of growth, namely the composition of the heterotrophic media. Upon transfer to ex vitro, the higher irradiances, compared to in vitro, can lead to oxidative stress symptoms, which can be counteracted by CO2 concentrations above atmospheric levels. Here we analyse the stability of Rubisco in in vitro grapevine plantlets, and after transfer to ex vitro under four acclimatization treatments: low irradiance (LL, 150 micromol m(-2)s(-1)) and high irradiance (HL, 300 micromol m(-2)s(-1)) in association with CO2 concentrations of 350 (LCO2) and 700 (HCO2) microL L(-1). Proteins were separated with SDS polyacrylamide gel electrophoresis and two-dimensional electrophoresis and Rubisco degradation peptides were analysed by immunoblotting with anti-LSU antibodies. These degradation products were present in the leaves of plantlets under both in vitro and ex vitro treatments. Under LCO2 they were maintained for almost all of the 28 days of the acclimatization period, while becoming scarcely detected after 14 days under HCO2 and after 7 days when HCO2 was associated with HL. These results appear to confirm the counteraction of HCO2 concentrations over the oxidative stress eventually caused by HL. The patterns of soluble sugars in acclimatizing leaves under HLHCO2 also gave an indication of a faster acquisition of autotrophic characteristics.

Carbon Dioxide↗

Characterisation of the ventilatory response to hypoxia in a model of transgenic anemic mice.

Both polycythemia and the increase in hypoxic ventilatory response (HVR) are considered as important factors of acclimatization to hypoxia. The objective of this study was to characterise the ventilation pattern at different inspired oxygen fraction in a model of chronic anemic mice. These mice have a targeted disruption in the 5' untranslated region of the Epo gene that reduces Epo expression such that the homozygous animal is severely anemic. Ventilation in normoxia in Epo-TAg(h) mice was significantly greater than in wild type, and the difference was mainly due to a higher tidal volume. HVR was higher in Epo-TAg(h) mice at every FIO2 suggesting a higher chemosensitivity. Resting oxygen consumption was maintained in anemic mice. Maximal oxygen consumption was 30% lower while hemoglobin was 60% lower in anemic mice compared to wild type. This small decrease in maximal oxygen consumption is probably due a greater cardiac output and/or a better tissue oxygen extraction and would allow these anemic mice to acclimatize to hypoxia in spite of low oxygen carrying capacity. In conclusion, Epo-TAg(h) anemic mice showed increased ventilation and hypoxic ventilatory response. However, whether these adaptations will contribute to acclimatization in chronic hypoxia remains to be determined.

Anemia↗

Disparate release of prolactin and growth hormone from the tilapia pituitary in response to osmotic stimulation.

In most teleost fishes, prolactin (PRL) plays a key role in freshwater (FW) adaptation, whereas growth hormone (GH) is involved in seawater (SW) adaptation in salmonids and certain euryhaline species including the tilapia, Oreochromis mossambicus. Consistent with its osmoregulatory activity, PRL release increases in response to physiologically relevant reductions in extracellular osmolality. When dispersed PRL and GH cells from FW-acclimatized fish were incubated in media of varying osmolalities, PRL release increased significantly in response to a 12% reduction in medium osmolality during 1 and 4h of exposure. By contrast, cells from SW-acclimatized fish responded only to a 24% reduction in osmolality. Growth hormone release on the other hand increased whether medium osmolality was reduced or raised. Cell volume increased together with PRL release during the perifusion of dispersed PRL cells in direct proportion to the reduction in medium osmolality. Growth hormone release increased whether GH cell volume increased or decreased. In in vivo studies, circulating PRL levels increased as early as 1h after the transfer of fish from SW to FW, whereas GH levels remained unchanged during 24h of acclimatization. These results indicate that while PRL and GH cells are osmosensitive, the PRL cells respond to reductions in extracellular osmolality in a manner that is consistent with PRL's physiological role in the tilapia. While the rise in GH release following the reduction in osmolality is of uncertain physiological significance, the rise in GH release with the elevation of medium osmolality may be connected to its role in SW adaptation.

Animals↗

Gene structure of the carp fish ribosomal protein L41: seasonally regulated expression.

The seasonal acclimatization of the carp fish demands physiological compensatory responses. The process involves profound nucleolar adjustments and remarkable changes in rRNA synthesis, which affect ribosomal biogenesis. We have documented that protein kinase CK2, whose activity is related to ribosomal protein L41 and the regulation of rRNA synthesis, was expressed in notably higher amounts in summer-acclimatized carp compared to the cold-season adapted fish. Thus, we approached the study of the functional genomics of carp L41 protein. We report the first cloning of a fish L41 gene encoding the highly conserved 25 amino acids, including approximately 1700 bp regulatory upstream region and the 3(') polyadenylation signal, plus the isolation and characterization of two different L41 cDNAs. We found a clear differential expression of L41, which follows the same pattern as protein kinase CK2beta that transcribes at higher levels in the summer-acclimatized carp than it does in the winter-adapted fish.

Animals↗

Effect of chromium(VI) on bacterial kinetics of heterotrophic biomass of activated sludge.

The effect of hexavalent chromium, Cr(VI), on the maximum specific growth rate, mu(m) and biomass yield, Y(H), of heterotrophic biomass was studied in batch tests conducted under high (= 10) and low (= 1.5) substrate-to-biomass ratios (S0/X0). The effects of sludge age and biomass acclimatization to Cr(VI) on the bacterial kinetics were also studied. The mu(m) values were determined by measuring oxygen uptake rate (OUR) and volatile suspended solids (VSS) increase. Cr(VI) concentrations equal or greater than 10 mg l(-1) inhibited the growth of unacclimatized activated sludge and caused a significant decrease in mu(m) and Y(H) values. The acclimatization of biomass and the selection of a high operating sludge age reduced the inhibitory effect of Cr(VI). At a sludge age of 20 days, Cr(VI) concentrations of <10 mg l(-1) stimulated bacterial growth as evidenced by an increase in both the mu(m) and Y(H) values. Determining mu(m) values by OUR and VSS methods, revealed that the presence of Cr(VI) in unacclimatized biomass caused an inhibitory effect mostly on substrate oxidation, while in acclimatized biomass, anabolic pathways were inhibited more.

Biomass↗

Production of polyhydroxyalkanoates (PHA) by activated sludge treating municipal wastewater: effect of pH, sludge retention time (SRT), and acetate concentration in influent.

In this paper, the production of biodegradable plastics polyhydroxyalkanoates (PHA) by activated sludge treating municipal wastewater was investigated. The effect of three operational factors, i.e. the acetate concentration in influent, pH, and sludge retention time (SRT) were studied. Sludge acclimatized with municipal wastewater supplemented with acetate could accumulate PHA up to 30% of sludge dry weight, while sludge acclimatized with only municipal wastewater achieved 20% of sludge dry weight. It was found that activated sludge with an SRT of 3 days possessed better PHA production capability than sludge with an SRT of 10 days. Sludge acclimatized under pH 7 and 8 conditions in sequencing batch reactors (SBRs) exhibited similar PHA production capability. However, in PHA production batch experiments, pH value influenced significantly the PHA accumulation behavior of activated sludge. When pH of batch experiments was controlled at 6 or 7, a very low PHA production was observed. The production of PHA was stimulated when pH was kept at 8 or 9.

Acetates↗

[Changes in spirometric parameters and arterial oxygen saturation during a mountain ascent to over 3000 meters].

OBJECTIVE: To ascertain whether climbing a mountain over 3000 meters high produces any alterations in ventilation, whether such alterations are modified by acclimatization, and whether they correlate with changes in arterial oxygen saturation (SaO2) or the development of acute mountain sickness (AMS). SUBJECTS AND METHODS: The following parameters were measured in 8 unacclimatized mountaineers who climbed Aneto (3404 m) and spent 3 days at the summit: forced vital capacity (FVC), forced expiratory volume in 1 second (FEV1), airway response to inhaled terbutaline, SaO2, and the symptoms of AMS. RESULTS: At the summit, mean (SD) FEV1 declined by 12.3% (5.7%) and mean FVC by 7.6% (6.7%) while the ratio of FEV1 to FVC remained normal. The means for both parameters were higher on the following day. No airway response to bronchodilator treatment was observed. The restriction disappeared entirely on descent. At the peak, SaO2 increased progressively as the climbers became acclimatized. During the ascent, FEV1 correlated with SaO2 (r=0.79). One participant who suffered from AMS had a ratio of FEV1 to FVC less than 70% and the worst SaO2 during the 3 days on the summit. Obstruction preceded the AMS symptoms, did not respond to bronchodilator treatment, and disappeared when the climber descended. CONCLUSIONS: The mountaineers who climbed over 3000 meters presented restriction that correlated with hypoxemia. This restriction did not respond to bronchodilator treatment, improved with acclimatization, and disappeared on descent. One person with AMS presented obstruction that did not respond to terbutaline and disappeared on descent.

Adult↗

The carp muscle-specific sub-isoenzymes of creatine kinase form distinct dimers at different temperatures.

We have previously cloned three muscle-specific sub-isoforms of creatine kinase (CK, EC 2.7.3.2) from the common carp ( Cyprinus carpio ), designated M1-CK, M2-CK, and M3-CK. The enzyme has a key role in maintaining the energy homoeostasis of cells with fluctuating energy requirements. In the present paper, we report that all three M-CKs in the red and white muscle of different temperature-acclimatized carp were ubiquitously distributed in the cytosol and along membranes. In addition, the expression levels of these isoforms were not significantly altered in response to the temperature acclimatization. Interestingly, our studies showed that the formation of distinct homo- or heterodimers among these three M-CKs was found at various temperatures. At higher temperature, the M1M1-CK and M2M2-CK homodimers, and the M1M3-CK heterodimer are the predominant MM-CKs, whereas the M3M3-CK achieves its homodimeric state at lower temperature. We postulated testable homology models to investigate the chemical properties of these dimeric interfaces. M1M1-CK was used as a reference to compare the structural differences with the M3M3-CK dimer. The calculated solvent accessible surface area that was buried in the contact interfaces of the M1M1-CK and M3M3-CK dimers showed an overall decrease of 12% in the M3M3-CK interface. The modelling analysis also suggested a net decrease of twelve hydrophobic residues and a Phe(3)-->Lys substitution in the M3M3-CK interface. An increase in thermolability of the M3M3-CK homodimer might be due to the decrease in subunit ion pairs and buried surface area in this dimer. Based on our findings, we propose that the carp-muscle-specific CK isoenzymes could undergo shuffling to form distinct M-CK homo- or heterodimers in acclimatization to environmental temperatures.

Amino Acid Sequence↗

Early steps in cold sensing by plant cells: the role of actin cytoskeleton and membrane fluidity.

Many plants acquire freezing tolerance through cold acclimatization (CA), a prolonged exposure to low but non-freezing temperatures at the onset of winter. CA is associated with gene expression that requires transient calcium influx into the cytosol. Alfalfa (Medicago sativa) cells treated with agents blocking this influx are unable to cold-acclimatize. Conversely, chemical agents causing increased calcium influx induce cold acclimatization-specific (cas) gene expression in alfalfa at 25 degrees C. How low temperature triggers calcium influx is, however, unknown. We report here that induction of a CA-specific gene (cas30), calcium influx and freezing tolerance at 4 degrees C are all prevented by cell membrane fluidization, but, conversely, are induced at 25 degrees C by membrane rigidification. cas30 expression and calcium influx at 4 degrees C are also prevented by jasplakinolide (JK), an actin microfilament stabilizer, but induced at 25 degrees C by the actin microfilament destabilizer cytochalasin D (CD). JK blocked the membrane rigidifier-induced, but not the calcium channel agonist-induced cas30 expression at 25 degrees C. These findings indicate that cytoskeleton re-organization is an integral component in low-temperature signal transduction in alfalfa cell suspension cultures, serving as a link between membrane rigidification and calcium influx in CA.

Actins↗

Deposition and clearance of labeled cigarette smoke particles in rat respiratory and digestive tract.

In this investigation 160 8-wk-old Sprague-Dawley rats inhaled smoke from 14C-labeled cigarettes in a Hamburg II smoking machine. The research cigarettes were labeled with [16,17-14C]dotriacontane. Eighty of the rats were acclimatized to smoke from nonlabeled research cigarettes for 21 d before inhaling labeled cigarette smoke. The investigation was divided into four experiments. In experiment I all rats were killed immediately after exposure to the 14C-labeled smoke from 30 cigarettes (10 muCi per cigarette) and the organs of the respiratory and digestive tracts were measured for 14C activity. These animals showed high activity in the apex nasi, fundus nasi, and larynx. The highest activity was found in the lobus dexter caudalis and the lobus sinister of the lung. The lungs contained four-fifths of the condensate inhaled into the respiratory tract. In experiment II three animals were killed immediately after smoking, three after 6 h, and four after 24 h. No activity was found after 6 h in apex, fundus nasi, larynx, and trachea. The lungs showed no clearance at all. The activity in the digestive tract was highest in the stomach and the esophagus immediately after cessation of smoking. These values decreased rapidly after 6 h. In experiment III the animals were killed after 7, 21, and 42 d. Clearance from the upper respiratory tract was fast, whereas clearance from the lungs was slow. Clearance from all lung lobes was faster in the nonacclimatized animals. In experiment IV the rats inhaled the smoke from 14C-labeled cigarettes (2 muCi per cigarette) on 5 successive days. The animals were killed 7, 14, and 21 d after the last smoke inhalation. Clearance from the lungs was slower in acclimatized and non-acclimatized animals than in experiment III.

Animals↗

Simultaneous removal of volatile organic compounds (VOCs) and nitrogen: batch test.

A batch test was conducted to investigate the effect of the chemical oxygen demand, (COD)/N ratio, nitrate, nitrite and temperature, on the microbial degradation of volatile organic compounds (VOCs), and the denitrification capability using VOCs-acclimatized and un-acclimatized cultures. The nitrite reduction rates differed with each reactor, as follows: 15.418 NO2-N mg MLVSS g(-1) h(-1) in the benzene and methanol (BM) reactor, 27.463 NO2-N mg MLVSS g(-1) h(-1) in the toluene and methanol (TM) reactor and 44.358 NO2-N mg MLVSS g(-1) h(-1) in the methanol (M) reactor. According to the COD/N ratio, the nitrate reduction rates of the BM and TM reactors acclimatized by VOCs changed in the ranges of 29.4-33.41 NO3-N mg MLVSS g(-1) day(-1) and 56.4-65.9 NO3-N mg MLVSS g(-1) day(-1), respectively. Thus, benzene was not effectively utilized as a carbon source. Conversely, toluene was utilized as a carbon source by the denitrifiers under substrate limited conditions. The specific denitrification rates were also greater in the TM reactor than those for both the substrate limited and unlimited conditions in the BM reactor.

Air Pollution↗

Plasma amino acids and excretion of protein end products by mice fed 10 or 40% soybean protein diets with or without dietary 2-acetylaminofluorene or N,N-dinitrosopiperazine.

Studies were conducted with 5- to 8-week-old male and female B6C3F1 mice to determine the influence of two carcinogens, 2-acetylaminofluorene (AAF) and N,N-dinitrosopiperazine (DNP), on plasma amino acid concentrations and on the excretion of lipids and nitrogenous metabolites. The carcinogens, AAF and DNP, were fed at concentrations of 0.25 and 0.05 g/kg of purified diet, respectively. Soybean protein constituted either 10 or 40% of the diet. Nutritional balances were measured over a 7-day period, after 7 days of acclimatization. Females ate less feed, gained less weight during acclimatization and excreted less fecal lipid as a percentage of intake than males. On the average, animals fed 40% protein consumed less total feed than those fed 10% protein. During acclimatization, DNP-fed animals ate and gained significantly less than controls. During week 2 DNP-fed animals gained significantly less than controls, although their feed intake was not significantly different. Fecal lipid excretion as a percentage of intake was significantly lower with carcinogens in the diet. The 40% protein diets increased lipid excretion in total and as a percentage of intake. With the exception of decreasing fecal lipid, AAF caused no consistent changes in feed intake, body weight, nitrogen (N) retention or N excretion. Neither carcinogen significantly influenced total fecal or urinary N, or the relative concentrations of the different forms of urinary N, when expressed as a percentage of N intake. Plasma ammonia rose with AAF feeding and plasma histidine rose with DNP feeding. Plasma concentrations of other amino acids were not changed consistently by either carcinogen. Feeding 40% protein caused a significant rise in plasma branched-chain amino acids, glycine and phenylalanine, and a significant decline in aspartate, threonine, serine, proline, citrulline, lysine and arginine.

2-Acetylaminofluorene↗

Role of airway receptors in altitude-induced dyspnea.

PURPOSE: The purpose of this study was to examine the role of airway receptors in respiratory-related sensations after ascent to altitude. METHODS: Ratings of respiratory-related sensations, perceived exertion and acute mountain sickness, heart rate, and peripheral oxygen saturation were recorded at rest and exercise in male and female subjects who had inhaled either aerosolized saline or saline with tetracaine after acute ascent to an altitude of 3500 m and after prolonged acclimatization of 18 d at altitudes between 4000 and 5000 m. RESULTS: Tetracaine had no effect on respiratory-related sensations at altitude either at rest or during exercise, and male and female subjects experienced similar respiratory-related sensations. Sensations of rapid breathing were experienced at rest after acute exposure to 3500 m as compared with sea level, but not after acclimatization to 5000 m. Sensations of rapid breathing, air hunger, and heavy breathing were experienced during exercise after acute and prolonged altitude exposure as compared with sea level, with a sensation of chest tightness experienced at 3500 m and a sensation of gasping experienced at 5000 m. CONCLUSION: These results suggest that airway afferents play no role in the respiratory-related sensations experienced by male and female subjects either during acute ascent to altitude or after prolonged acclimatization at altitude.

Adult↗

Influence of two beta-adrenoceptor antagonists, propranolol and pindolol, on cold adaptation in the rat.

1. Adult male rats were treated with propranolol (2.0 mg kg-1 day-1 i.p.), pindolol (0.2 mg kg-1 day-1 i.p.) or 0.9% NaCl day-1 i.p. and exposed to +4 degrees C for 42 days, or treated with 0.9% NaCl day-1 i.p. and kept at +23 degrees C for 42 days. They were weighed once a week, when a 24 h urine sample was also collected and colon temperature measured. 2. Urinary noradrenaline (NA), adrenaline (Ad) and dopamine were analysed by high-performance liquid chromatography with an electrochemical detector. After the acclimatization period the interscapular brown adipose tissue was excised and weighed and the activity of the oxidative enzymes succinate dehydrogenase and cytochrome oxidase measured. 3. The pindolol-treated and propranolol-treated rats gained weight during the cold-acclimatization period. The amount of interscapular brown adipose tissue increased in the cold, but the increase was lowest in the pindolol-treated group. No changes were seen in the other brown adipose tissue parameters in cold-exposed animals. The excretion of catecholamines followed the same pattern in all three cold-exposed groups, with an initial rise in noradrenaline and adrenaline excretion and a slight rise in dopamine excretion. 4. The results suggest possible connections between beta-adrenoceptor antagonists, weight gain and cold acclimatization. Pindolol had a slight inhibitory effect on cold-induced brown adipose tissue hypertrophy in rats.

Adaptation, Physiological↗