Effects of experimental ventilation and ambient Po2 on O2 uptake of the isolated cutaneous tissue in the carp,Cyprinus carpio

Effects of experimental ventilation and ambient Po₂ on cutaneous O₂ uptake in vitro were studied in the carp, Cyprinus carpio. Oxygen uptake rate of the isolated cutaneous tissue was determined by ventilating the epidermis side of the skin with normoxic water in flow-through respirometers. Oxygen uptake rate of the skin increased with ventilation rate across the skin between 2.5 and 40 ml/min and became 3.2 nmol/cm²/min at a flow rate of 40 ml/min, which corresponds to an apparent water velocity of 1.1 cm/sec. At a ventilation rate of 10 ml/min, oxygen uptake rate of the skin increased with the ambient Po₂ between 115 and 230 Torr and became constant (3.8 nmol/cm²/min) between 230 and 295 Torr. When both sides of the skin were ventilated with normoxic water, oxygen uptake rate of the skin increased and became 3.7 nmol/cm²/min at a flow rate of 20–40 ml/min. These results suggest that the oxygen requirement of the skin is 3.7–3.8 nmol/cm²/min at 21.3°C and that cutaneous O₂ uptake in vitro depends on experimental ventilation and ambient Po₂, consistent with values measured in vitro in the carp (ref).     

28-Homobrassinolide ameliorates the saline stress in chickpea (Cicer arietinum L.)

The response of chickpea (Cicer arietinum L.) cv. KPG-59 to pre-sowing seed treatment with 28-homobrassinolide (HBR) and/or sodium chloride (NaCl) was investigated. The seeds imbibed in aqueous solution of 10⁻¹⁰ or 10⁻⁸ M of HBR for 8 h, resulted in an increase in the values for most of the characteristics of shoot and root at 90-day stage and seed yield, at harvest. The plants resulting from the seeds soaked in HBR (10⁻⁸ M) possessed 23% and 31% higher leaf nitrate reductase (E.C. 1.6.6.1) and carbonic anhydrase (E.C. 4.2.2.1) activities, 34% more dry mass, 30% higher nodule number, 31% and 18% more nodule fresh and dry mass, compared with water soaked, control. Leghaemoglobin content and nitrogenase activity (E.C. 1.7.99.2) were 28% and 30% higher while nodule nitrogen and carbohydrate contents decreased by 5% and 6%, compared with the control. Moreover, seed yield increased by 26% over the control, at harvest. The values for all the above characteristics declined significantly, in the plants raised from the seeds soaked in NaCl. However, this ill effect was overcome, if NaCl treatment was given before or after HBR treatment.     

Increased oxygen consumption during the uptake of water by the eversible vesicles of Petrobius brevistylis

The oxygen consumption of individuals of Petrobius brevistylis has been measured before, during, and after the uptake of fluid by the insects eversible abdominal vesicles. There was no difference in consumption between fresh animals and those that had been dehydrated or those that had been allowed to rehydrate after an exposure to a NaCl solution. However, during the process of taking up fluid there was over a 200% increase in the respiration rate. While the increase in the rate of oxygen consumption was related to the concentration of the fluid from which the animals were taking up water, the increase in the amount of oxygen consumed was related to the concentration of the substrate solution and the amount of uptake. Part of the oxygen increase is ascribed to the posture adopted by the animals during the uptake of fluid and the remainder to the active transport processes that may be occurring.     

Myelosuppression in the Pig (Sus scrofa): Uteroferrin Reduces the Myelosuppressive Effects of 5-Fluorouracil in Young Pigs

The present study investigated the ability of uteroferrin to modulate the myelosuppressive effects of 5-fluorouracil (5-FU) in young pigs (Sus scrofa). Pigs (28–35 days of age; n = 6 per treatment) were infused with equal amounts of 5-FU on days 0 and 1 of the experimental period (37.5 mg/kg cumulative dose). Uteroferrin (100 μg/kg in 0.9% NaCl) or control (equivalent volume of 0.9% NaCl) was administered to pigs as intramuscular injections twice daily (08:00 and 20:00 hr) on days 1 through 21. Peripheral blood cell number, composition and progenitor cells were determined over 28 days. Treatment of pigs with 5-FU resulted in a rapid dose-dependent (P < 0.05) leukocytopenia. Concurrent treatment of pigs with uteroferrin reduced (P < 0.05) the rate of 5-FU-induced leukocytopenia (44 vs 77 ± 7% decline from baseline on day 3) and enhanced (P < 0.05) the recovery from 5-FU on days 10 and 12 postinfusion. The positive effect of uteroferrin on leukocytes resulted primarily from a protection and/or enhanced recovery of neutrophils and monocytes. In addition, uteroferrin attenuated (P < 0.05) the suppression of red blood cell numbers after 5-FU administration (6.9 vs 6.1 ± 0.2 × 10⁶ cells/μl on day 3), an affect reflected in increased hematocrit and hemoglobin concentrations. The effects of uteroferrin appeared to result from enhancement of the proliferation and/or differentiation of primitive pluripotent stem cells resistant to 5-FU, as concurrent treatment of pigs with uteroferrin resulted in a protection and/or enhanced recovery (P < 0.05) of CFU-GEMM, CFU-GM and BFU-E progenitor cells in the peripheral blood. These results are the first to demonstrate that uteroferrin can reduce the myelosuppressive effects of 5-FU in the pig and suggest that uteroferrin has hematopoietic growth factor activity in vivo.     

Effect of temperature on electrolyte metabolism of isolated frog skin

A study is presented on the effect of temperature on unidirectional active ion transport, resting electrolyte equilibrium (electrolyte composition), and oxygen consumption in isolated frog skin. The aims were twofold: first, to find out whether the rate of active transport can be changed without affecting the Na⁺ and K⁺ balance of skin itself; second, to arrive at minimal ΔNa/ΔO₂ values by correlating quantitatively inhibition of active ion transport with inhibition of O₂ consumption. NaCl transport was maximal at 20°C. At 28° and at temperatures below 20°, rate of NaCl transport was diminished. In many instances NaCl transport was diminished in skins which maintained their normal Na⁺ and K⁺ content. In several cases, however, neither rate of transport nor resting electrolyte equilibrium was affected; in other cases, both were. O₂ consumption decreased when lowering the temperature over the range from 28 to 10°C. From a plot of log Q_OO2 against 1/T an activation energy of µ 13,700 cal. was calculated, valid for the range from 10 to 20°C. It appeared that µ was smaller for temperatures above 20°C. Working between 10 and 20°, it was found that, on the average, 4 to 5 equivalents of Na⁺ were transported for one mole of O₂ consumed in skins with undisturbed resting electrolyte equilibrium.     

Metabolism of excised rat skin in hypertonic media

The rate of oxygen consumption by rat skin is inhibited similarly by hypertonic solutions of NaCl, KCl, and sucrose. The rate of anaerobic glycolysis is inhibited by hypertonic NaCl. The inhibition of QO₂ by NaCl concentrations above about 2 osmols per liter is not reversible.     

Respiratory metabolism of the semen of the honey-bee, Apis mellifera

Mean respiratory quotients determined on samples from individual drones by Cartesian diver respirometry averaged 0·64, suggesting that phospholipids constitute the major source of energy. The average sperm density of semen was 7·76 millions/μl. Dilution of semen increased the rate of oxygen consumption by 68 per cent. Semen stored at 13 to 14°C for 1 month showed only 40 per cent of the oxygen uptake of freshly ejaculated semen. Streptomycin sulphate treated samples of semen showed significantly lower rates of oxygen consumption than untreated samples. These results suggest that the high density of sperm in the semen and low metabolic activity during storage may be at least partly responsible for the successful long-term storage of honey-bee spermatozoa.     

Action du diméthylsulfoxyde sur l'absorption du 42K et du glucose-14C (U) par des disques foliaires de Zea mays et Pelargonium zonale

⁴²K and ¹⁴C (U)-glucose uptake by foliar discs of maize (Zea mays L.) and pelargonium (Pelargonium zonale L.) is inhibited by dimethyl sulfoxide (DMSO) at concentrations of 5 and 10%. This effect increases with the duration of the absorption and appears with varied concentrations in K or glucose. In most cases, this inhibitory effect is less with aged discs than with freshly cut discs. Besides, the DMSO (10%) enhances the exsorption of ⁴²K by foliar discs of maize. The effects of DMSO and two osmotic agents, mannitol and polyethylene glycol 1000, on the uptake and water content of discs are compared. It seems that the effect of DMSO on the uptake is not only an osmotic effect. It may also be related to other actions of this substance in affecting an active uptake.     

Failure to detect nephrotoxicity of chronically administered dimethyl sulfoxide (DMSO) in rats.

Male and female rats received daily intraperitoneal injections of dimethyl sulfoxide (DMSO; 2 or 4 g/kg) or saline for 28 days. At the end of that time, the ability of the animals' kidneys to transport p-aminohippurate (PAH) and N-methylnicotinamide (NMN) was tested in an in vitro system.Both doses of DMSO used produced measurable toxicity of a general nature: There was some mortality, and growth of the animals was retarded. However, DMSO had no detectable nephrotoxic action. Terminal serum urea and creatinine concentrations were normal in all treatment groups, and there was no effect of DMSO on the uptake of PAH or NMN by renal cortical slices. The data suggest that DMSO does not have an important specific toxic effect on the kidneys in this species.     

K+--induced changes of oxygen uptake by neuronal enriched and glial enriched fractions from mouse brain cortex.

Neuronal and glial enriched fractions were incubated in a medium with 10mM pyruvate, 5mM fumarate and 0.9mM 5'-AMP and the effect of increased external K+ concentrations was studied upon oxygen uptake. A concentration of 65 mM K+ had a different effect on the oxygen consumption of glial and neuronal perikarya. The rate of oxygen uptake by glia was stimulated by 52.81% whilst an insignificant decrease of 15.79% occurred in the neurones. The highest rate of oxygen uptake by incubated cells was estimated in the presence of the substrate system containing pyruvate, fumarate and 5'-AMP. The significance of components in the substrate system for a high rate of oxygen uptake by cells was also tested with 6.2 mM K+ and 65 mM K+.     

L-serine and GABA uptake by synaptosomes during postnatal development of rat

Postnatal development changes in mechanisms of synaptosomal amino acid transport have been studied in rat cerebral cortex. Specific uptake of radiolabeled L-serine was examined and compared with that of radiolabeled GABA using synaptosomes-enriched fractions freshly prepared from cerebral cortex at different postnatal days from the birth to young adulthood. The preparations were incubated with 10 nM of [3H]L-serine and 10 nM of [3H]-GABA in either the presence or absence of NaCl, KCl or choline chloride, at 2 and 30 degrees C, for different periods up to 30 min. The uptake of [3H]l-serine was temperature dependent in synaptosomal fractions prepared from cerebral cortex of rats in postnatal days 5, 7, 13 and 21, but stronger dependence was observed in adult brain, irrespective of the presence of Na+, K+ or choline ions. At all postnatal ages studied, [3H]-GABA uptake showed a high activity in the presence of Na+ ions and at 30 degrees C. The values of Km were 90-489 microM in L-serine uptake. However, in the uptake of GABA the values of Km were 80-150 microM. The highest values of Vmax were obtained at 5 and 21 postnatal days for both transport systems. These results indicate that the uptake of l-serine and GABA are regulated differentially during postnatal development.     

Body temperature, heart rate and oxygen consumption of normothermic and heterothermic western jumping mice (Zapus princeps)

1. Heart rates and oxygen consumption were measured for normothermic, hibernating, and arousing Zapus princeps. 2. Rectal and oral body temperature were monitored and differential rates of rewarming of the body were recorded. 3. Lengths of periodic arousals differ in two stages of hibernation; days 50-80 arousal durations were 12 hr, 43 min while days 140-210 were 10 hr, 28 min. 4. Total energy budget of hibernation was calculated for a 295 day hibernation cycle based on the utilization of 18,020 cc of oxygen for that period. The calculated utilization of fat would be 8.97 g while the observed use was 9.5-11.1 g.     

A novel oxyconforming response in the freshwater fish Galaxias maculatus

How fish oxygen consumption is modulated by external PO₂ has long been a matter of interest, yet is an experimentally complicated question to answer. In this study closed and semi-closed respirometry were used to evaluate the oxygen consumption rate of the scaleless galaxiid fish, inanga (Galaxias maculatus) as a function of decreasing external PO₂. Both respirometry techniques showed that as environmental oxygen levels declined, oxygen consumption rates also decreased. At no point did inanga regulate oxygen consumption. This is strong evidence that inanga is an oxyconformer. Partitioned respirometry experiments showed that skin plays an important role in oxygen uptake in this fish species, and cutaneous oxygen uptake may have an important role in shaping the oxygen consumption response to hypoxia.     

Altered metabolic response of iron-deficient women during graded, maximal exercise.

Metabolic responses during a standardized, progressive, maximal work capacity test on a cycle ergometer were studied in 11 women, mean age 28 (SEM 2) years, at admission to the study, after their body iron stores were depleted by diet, phlebotomy and menstruation for about 80 days and after iron repletion by diet for about 100 days, including daily iron supplementation (0.9 mmol iron as ferrous sulfate) for the last 14 days of repletion. Iron depletion was characterized by a decline (P less than 0.05) in hemoglobin, ferritin and body iron balance. Iron repletion, including supplementation, increased (P less than 0.05) hemoglobin, ferritin and iron balance. No changes were observed in cardiovascular and ventilatory responses or peak oxygen uptake. Iron depletion was associated with a reduced (P less than 0.05) rate of oxygen utilization, total oxygen uptake and aerobic energy expenditure, and elevated (P less than 0.05) peak respiratory exchange ratio and post-exercise concentration of lactate. Reduction of body iron stores without overt anemia affects exercise metabolism by reducing total aerobic energy production and increasing glycolytic metabolism.     

Effect of sodium fluoride on radiation sensitivity of barley seeds

Barley seeds soaked in 0.01 M sodium fluoride (NaF) in phosphate buffer (pH7) or in buffer alone for 18 h were dried and equilibrated to 10% moisture, either in air or in nitrogen. Pre-treated and re-dried seeds were irradiated in air or in nitrogen with 0, 13, 20, 26 and 32 kR of γ rays, and were immediately hydrated in oxygen- or nitrogen-bubbled water. Parameters of radiation effect considered were seedling injury, mitotic and meiotic cells with bridge aberrations at anaphase and pollen fertility in M₁, and the frequency of chlorophyll mutations in M₂. NaF at 0.01 M was not mutagenic by itself. Pre-treatment with NaF significantly enhanced the radiation effect, when the irradiation was done in air, in comparison with the buffer soaked seeds. The increased effect due to NaF was additional to the oxygen effect. In nitrogen, NaF pre-treatment increased the mutagenic effect but it was not always significant. Post-soaking of irradiated seeds in 0.01 M NaF for 5 h increased seedling injury in comparison with the irradiated seeds soaked in buffer alone or in 0.01 M NaCl. At least a part of the sensitizing effect of NaF may be due to the inhibition of repair.     

ON THE EFFECTS OF DMSO IN CATION TRANSPORT BY EXCISED BARLEY ROOTS

Dimethyl sulfoxide (DMSO) in concentrations of up to 10% by volume stimulates the uptake of zinc by excised barley roots. In the same concentration it severely depresses uptake of sodium and of rubidium. It does not seem to affect the permeability of the membrane since roots treated with desorption solutions which were 10% in DMSO did not lose more of the preferred ion than did roots desorbed in solutions not containing DMSO. Oxygen utilization (measured in the Warburg respirometer) was reduced when DMSO was present. It is suggested that DMSO is a poisoning agent which interferes with cation transport by attacking some aspect of metabolism and not by influencing the permeability of the membrane.     

Respiratory and osmoregulatory responses of the hermit crab, Clibanarius vittatus (Bosc), to salinity changes

Intertidal hermit crabs were stepwise acclimated to 10, 20, and 30‰ salinity (S) and 21 ± 1 °C. Hemolymph osmolality, sodium, chloride, and magnesium were isosmotic (isoionic) to ambient sea water at 30‰ and hyperosmotic (hyperionic) at 20 and 10‰ S, while hemolymph potassium was significantly hyperionic in all acclimation salinities. Total body water did not differ significantly at any acclimation salinity. Oxygen uptake rates were higher in summer-than winter-adapted crabs. No salinity effect on oxygen consumption occurred in winter-adapted individuals. Summer-adapted, 30‰ acclimated crabs had a significantly lower oxygen consumption rate than those acclimated 10 and 20‰ S. Crabs exposed to 30 10 30‰ and 10 30 10‰ semidiurnal (12 h) and diurnal (24.8 h) fluctuating salinity regimes showed variable osmoregulatory and respiratory responses. Hemolymph osmolality followed the osmolality of the fluctuating ambient sea water in all cases, but was regulated hyperosmotically. Hemolymph sodium, chloride, and magnesium concentrations were similar to hemolymph osmolality changes. Sodium levels fluctuated the least. Hemolymph potassium was regulated hyperionically during all fluctuation patters, but corresponded to sea water potassium only under diurnal conditions. The osmoregulatory ability of Clibanarius vittatus (Bosc) resembles that reported for several euryhaline brachyuran species. The time course of normalized oxygen consumption rate changed inversely with salinity under semidiurnal and diurnal 10 30 10‰ S fluctuations. Patterns of 30 10 30‰ S cycles had no effect on oxygen consumption rate time course changes. The average hourly oxygen consumption rates during both semidiurnal fluctuations were significantly lower than respective control rates, but no statistical difference was observed under diurnal conditions.     

Cryopreservation of cultured mantle cells of Paphia malabarica for perennial availability

Laboratory friendly, cryopreservation procedures with respect to cryopreservation formulations and cryopreservation temperatures were attempted, in the present study to ensure perennial availability of cultured mantle cells of bivalve (Paphia malabarica). Screening of cryopreservative formulations with different concentrations of DMSO, Propylene glycol and Glycerol was carried out for cryopreservation of freshly dissociated cells of Paphia malabarica. Out of these cryopreservative formulations, 10% DMSO, 10% Propylene glycol and 15% Glycerol were selected for cryopreservation of the mantle cells pooled from 1-day old primary culture and cell line after 3 passages at the end of different cryopreservation periods. Cryopreservative formulation with 15% glycerol, served as a best cryoprotectant for the cryopreservation of cells sourced from freshly dissociated cells as well as from primary cultures and cell cultures after three passages of mantle cells of Paphia malabarica, retaining metabolic activity of resurrected cells. Both, cell cultures established from uncryopreserved cells as well as cryopreserved cells showed similar alkaline phosphatase and carbonic anhydrase activities thus indicating retention of their biomineralization capacity even after cryopreservation at low and ultralow temperatures.     

Oxygen consumption by the bivalve Donax vittatus (da Costa)

The acute oxygen consumption of Donax vittatus (da Costa) freshly collected at different times from a beach at Barrassie, Ayrshire, Scotland, has been measured at different temperatures. The logarithmic relationship between oxygen consumption and body weight showed a significant difference on only one occasion, and a common regression coefficient (b) of 0.865 could be used for regressions of oxygen consumption on weight. Over the temperature range 2.9–20 °C oxygen consumption rose with temperature. There was a linear decline of Q₁₀ with temperature in the range 2.9 –20 °C. Differences in values of the constant (a) in the regression equation suggest that there is some acclimation to temperature, resulting in rotation of the rate/temperature curve counterclockwise for warm-acclimated animals, and a reduction of Q₁₀ in cold-acclimated animals. The differences in oxygen consumption which result are small and appear to have little practical significance. High levels of metabolically-inactive materials such as stored glycogen reserves lead to a reduction in the weight-specific oxygen consumption. Spawning animals show an increased oxygen consumption.     

Viability and engraftment of hematopoietic progenitor cells after long-term cryopreservation: effect of diagnosis and percentage dimethyl sulfoxide concentration

Background aimsWe carried out a retrospective analysis of viability by diagnosis and dimethyl sulfoxide (DMSO) concentration in patients who had undergone autologous transplants using hematopoietic progenitor cells (HPC) after long-term storage (up to 17.8 years).MethodsViability was tested using flow cytometry for HPC that were harvested and preserved using a controlled rate freezer and 5% or 10% DMSO with human serum albumin, then stored in liquid nitrogen. Data from 262 samples were analyzed (249 myeloma patients and 13 other diagnoses): 100 consecutively thawed samples with a storage time of <1 year (all 10% DMSO), 50 consecutive samples stored for 1–4.9 years (10% DMSO), 50 samples stored for 5–9 years (5% DMSO) and all samples stored and used for transplant after >9 years (60 samples, 5% DMSO; two samples, 10% DMSO).ResultsNo statistically significant difference in viability between the 5% DMSO and 10% DMSO groups was observed (P = 0.08), so the 1–4.9 years and 5–9 years were combined and the three groups (<1 year, 1–9 years and >9 years) were compared using an anova test. There was no difference in viability based on cryostorage period (P = 0.23) or between myeloma and other diagnoses (P = 0.45). No difference was seen in time to White blood cell (WBC) engraftment (P = 0.10) or to platelet engraftment between groups (P = 0.52).ConclusionsThese data suggest that long-term storage in 5% DMSO and human serum albumin is safe.