Joaquim Azcón-Bieto

The metabolic environment of cancer

The tumor cell has a very distinctive metabolism. It acts as a metabolic trap for host nutrients thus taking vital compounds for the metabolism of the host. Depending on the particular tumor growing pattern, cancer cells use preferentially glucose or amino acids for their energetic or biosynthetic needs. Lipids, fatty acids in particular, can also be taken up by the tumor cell. In addition, it can also release some compounds into the host circulation which are not normally produced by the original cell before neoplastic transformation. Some of these compounds affect the metabolism of the host in an unfavorable way since they can oppose the hosts metabolic responses, which sustain homeostasis. The final product is that the metabolic machinery of these cells allows them to grow continuously in an uncontrolled manner. The consequences of tumor invasion on the hosts metabolism are varied. They have, however, one thing in common: the reduction of the metabolic efficiency of the host. Muscular protein depletion, increased gluconeogenesis, uncoupling of oxidative phosphorylation constitute the main metabolic responses of the host as a result of tumor invasion. The net result of all these metabolic changes is profound energy imbalance which normally ends with cachexia and, eventually, death.

Acclimation of Respiratory O2 Uptake in Green Tissues of Field-Grown Native Species after Long-Term Exposure to Elevated Atmospheric CO2

C3 and C4 plants were grown in open-top chambers in the field at two CO2 concentrations, normal ambient (ambient) and normal ambient + 340 [mu]LL-1 (elevated). Dark oxygen uptake was measured in leaves and stems using a liquid-phase Clark-type oxygen electrode. High CO2 treatment decreased dark oxygen uptake in stems of Scirpus olneyi (C3) and leaves of Lindera benzoin (C3) expressed on either a dry weight or area basis. Respiration of Spartina patens (C4) leaves was unaffected by CO2 treatment. Leaf dry weight per unit area was unchanged by CO2, but respiration per unit of carbon or per unit of nitrogen was decreased in the C3 species grown at high CO2. The component of respiration in stems of S. olneyi and leaves of L. benzoin primarily affected by long-term exposure to the elevated CO2 treatment was the activity of the cytochrome pathway. Elevated CO2 had no effect on activity and capacity of the alternative pathway in S. olneyi. The cytochrome c oxidase activity, assayed in a cell-free extract, was strongly decreased by growth at high CO2 in stems of S. olneyi but it was unaffected in S. patens leaves. The activity of cytochrome c oxidase and complex III extracted from mature leaves of L. benzoin was also decreased after one growing season of plant exposure to elevated CO2 concentration. These results show that in some C3 species respiration will be reduced when plants are grown in elevated atmospheric CO2. The possible physiological causes and implications of these effects are discussed.

The reaction of the plant mitochondrial cyanide-resistant alternative oxidase with oxygen

The dependence of electron flux through the cyanide-resistant respiratory pathway on the redox poise of the ubiquinone pool and oxygen concentration was studied in purified mitochondria isolated from green and etiolated soybean (Glycine max L. Merr. cv. Ransom) cotyledons at different ages (4 and 10 days after planting), soybean roots and mung bean (Vigna radiata L.R. Vilcz) hypocotyls. In soybean, the Km of the alternative oxidase with respect to oxygen was found to vary between values of 10 and 20 μM. These are generally higher than values of the Km for oxygen of the alternative oxidase reported previously (0.5 to 2.0 μM). In addition, the value of the Km for oxygen varied with the redox poise of the ubiquinone pool, measured voltametrically; the more reduced the quinone pool, the larger the observed Km. These results are at variance with the behavior expected of the kinetic model developed by Siedow and Moore (1993; Biochim. Biophys. Acta 1142, 165–174) which predicts that the Km for oxygen should decrease as the quinone pool becomes more reduced. A modified kinetic model is developed that incorporates an additional reaction step involving activation of the four-electron reduced oxidase into the earlier kinetic model. The modified model successfully simulates the dependence of the alternative oxidase activity on both ubiquinone pool redox poise and oxygen concentration.

Tumour necrosis factor-alpha uncouples respiration in isolated rat mitochondria

Recent studies have demonstrated the existence of an intracellular (associated with mitochondria) tumour necrosis factor-alpha (TNF) binding protein. In an attempt to elucidate if this receptor could be involved in TNF action, we have incubated liver isolated mitochondria in the presence of recombinant murine TNF. The results show that the addition of TNF at concentrations as low as 10(-6) U/microl resulted in a clear uncoupling respiration of liver isolated mitochondria, therefore suggesting that TNF can indeed exert intracellular effects, which are possibly linked with its cytotoxic mechanism.

Changes in Respiratory Mitochondrial Machinery and Cytochrome and Alternative Pathway Activities in Response to Energy Demand Underlie the Acclimation of Respiration to Elevated CO2 in the Invasive Opuntia ficus-indica

Studies on long-term effects of plants grown at elevated CO2 are scarce and mechanisms of such responses are largely unknown. To gain mechanistic understanding on respiratory acclimation to elevated CO2, the Crassulacean acid metabolism Mediterranean invasive Opuntia ficus-indica Miller was grown at various CO2 concentrations. Respiration rates, maximum activity of cytochrome c oxidase, and active mitochondrial number consistently decreased in plants grown at elevated CO2 during the 9 months of the study when compared to ambient plants. Plant growth at elevated CO2 also reduced cytochrome pathway activity, but increased the activity of the alternative pathway. Despite all these effects seen in plants grown at high CO2, the specific oxygen uptake rate per unit of active mitochondria was the same for plants grown at ambient and elevated CO2. Although decreases in photorespiration activity have been pointed out as a factor contributing to the long-term acclimation of plant respiration to growth at elevated CO2, the homeostatic maintenance of specific respiratory rate per unit of mitochondria in response to high CO2 suggests that photorespiratory activity may play a small role on the long-term acclimation of respiration to elevated CO2. However, despite growth enhancement and as a result of the inhibition in cytochrome pathway activity by elevated CO2, total mitochondrial ATP production was decreased by plant growth at elevated CO2 when compared to ambient-grown plants. Because plant growth at elevated CO2 increased biomass but reduced respiratory machinery, activity, and ATP yields while maintaining O2 consumption rates per unit of mitochondria, we suggest that acclimation to elevated CO2 results from physiological adjustment of respiration to tissue ATP demand, which may not be entirely driven by nitrogen metabolism as previously suggested.

Actual and potential dark respiration rates and different electron transport pathways in freshwater aquatic plants

Abstract The rates of respiratory O 2 uptake have been studied in leaves, stems and whole shoots of several freshwater plants: 6 angiosperms, 2 bryophytes and one alga. For angiosperm leaves, rates varied widely with species (30–142 μmol O 2 (gDW) −1 h −1 ), were correlated with chlorophyll content and were higher than those of the stems (13–71 μmol O 2 (gDQ) −1 h −1 ). The rates for the shoots of bryophytes (53–66 μmol O 2 (gDW) −1 h −1 ) and for the alga Cladophora glomerata (L.) Kutz. (96 μmol O 2 (gDW) −1 h −1 ) were slightly higher than those of most angiosperm stems, but lower than those for most leaves. These plants had a significant cyanide-resistant respiration, suggesting the existence of an alternative pathway to the “classic” cytochrome system. This pathway was found to be active in all the species studied, as judged by responses to a specific inhibitor, SHAM (salicylhydroxamic acid). Measurement of electron-transport system (ETS) activity showed that there is a large electron-transport capacity which is not normally used by respiration in vivo.

Assessing the stable carbon isotopic composition of intercellular CO2 in a CAM plant using gas chromatography-combustion-isotope ratio mass spectrometry

Most of the literature focused on internal CO(2) (Ci) determinations in plants has used indirect methods based on gas-exchange estimations. We have developed a new method based on the capture of internal air gas samples and their analysis by gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS). This method provided a direct measure of intercellular CO(2) concentrations combined with stable carbon isotopic composition in O. ficus-indica plants. Plants were grown at both ambient and elevated CO(2) concentration. During the day period, when the stomata are closed, the Ci was high and was very (13)C-enriched in both ambient and elevated CO(2)-grown plants, reflecting Rubiscos fractionation (this plant enzyme has been shown to discriminate by 29 per thousand, in vitro, against (13)CO(2)). Other enzyme fractionations involved in C metabolism in plants, such as carbonic anhydrase, could also be playing an important role in the diurnal delta(13)C enrichment of the Ci. During the night, when stomata are open, Ci concentrations were higher in elevated (and the corresponding delta(13)C values were more (13)C-depleted) than in ambient CO(2)-grown plants.

The impairment of respiration by glycolysis in the Lewis lung carcinoma

Isolated vegetative tumour cells from mice bearing the Lewis lung carcinoma showed low rates of basal respiration with both low oxygen uptake rates and cytochrome-c oxidase activity. The cells were affected by a marked Crabtree effect and a high rate of lactate production in the presence of 10 mM glucose. The glycolytic capacity of the tumour was also assessed through the measurement of the maximum activities for hexokinase, phosphofructokinase, pyruvate kinase and lactate dehydrogenase. These activities were similar to the ones found in other fast-growing, undifferentiated tumours. The concentration of fructose-2,6-bisphosphate in the tumour was 2,3 nmoles/g fresh tissue wt., a value which is of the same order of magnitude as that found in other types of highly glycolytic cells. It is concluded that the Lewis lung carcinoma follows the same pattern as other undifferentiated tumours with a high capacity for both glucose and amino acid utilization.

Effects of long‐term exposure to elevated CO2 conditions in slow‐growing plants using a 12C‐enriched CO2‐labelling technique

Despite their relevancy, long-term studies analyzing elevated CO(2) effect in plant production and carbon (C) management on slow-growing plants are scarce. A special chamber was designed to perform whole-plant above-ground gas-exchange measurements in two slow-growing plants (Chamaerops humilis and Cycas revoluta) exposed to ambient (ca. 400 micromol mol(-1)) and elevated (ca. 800 micromol mol(-1)) CO(2) conditions over a long-term period (20 months). The ambient isotopic (13)C/(12)C composition (delta(13)C) of plants exposed to elevated CO(2) conditions was modified (from ca. -12.8 per thousand to ca. -19.2 per thousand) in order to study carbon allocation in leaf, shoot and root tissues. Elevated CO(2) increased plant growth by ca. 45% and 60% in Chamaerops and Cycas, respectively. The whole-plant above-ground gas-exchange determinations revealed that, in the case of Chamaerops, elevated CO(2) decreased the photosynthetic activity (determined on leaf area basis) as a consequence of the limited ability to increase C sink strength. On the other hand, the larger C sink strength (reflected by their larger CO(2) stimulatory effect on dry mass) in Cycas plants exposed to elevated CO(2) enabled the enhancement of their photosynthetic capacity. The delta(13)C values determined in the different plant tissues (leaf, shoot and root) suggest that Cycas plants grown under elevated CO(2) had a larger ability to export the excess leaf C, probably to the main root. The results obtained highlighted the different C management strategies of both plants and offered relevant information about the potential response of two slow-growing plants under global climate change conditions.

Water status, photosynthetic pigments, C/N ratios and respiration rates of Sitka spruce seedlings exposed to 70 ppbv ozone for a summer

Abstract Three-year old seedling of Picea sitchensis (Bong.) Carr. were exposed in large-scale chambers (solardomes) to 70 ppbv O 3 for an entire summer, 7 hr day −1 , under approximately ambient conditions. No macro- or microscopic injury, no fluorescence differences and no growth rate effects were found in the fumigated spruce. However, fumigated seedlings showed significantly lowered water potentials, chlorophyll concentrations, nitrogen and sulfur contents, and respiration rates. The old needles had higher cyanide-resistant respiration. These results were consistent with symptoms of forest decline affecting high-altitude forest and showed the presence of physiological changes even though there were no visible symptoms.