Inhibition of nitrification by waste tea (‘Tea Fluff’)

Summary Ammonium fertilizer applied to tea soils is readily converted to nitrate by the nitrifying bacteria in soil. Excess nitrate in soil could undergo rapid leaching losses under high rainfall conditions. Data is presented in this paper to show that waste tea could be effectively used to retard and delay nitrate production and thereby prevent loss of nitrogen as nitrate by leaching. Evidence is also presented to show that waste tea readily liberates ammonium nitrogen in soil.     

Does canopy nitrogen uptake enhance carbon sequestration by trees?

Temperate forest ¹⁵N isotope trace experiments find nitrogen (N) addition‐driven carbon (C) uptake is modest as little additional N is acquired by trees; however, several correlations of ambient N deposition against forest productivity imply a greater effect of atmospheric nitrogen deposition than these studies. We asked whether N deposition experiments adequately represent all processes found in ambient conditions. In particular, experiments typically apply ¹⁵N to directly to forest floors, assuming uptake of nitrogen intercepted by canopies (CNU) is minimal. Additionally, conventional ¹⁵N additions typically trace mineral ¹⁵N additions rather than litter N recycling and may increase total N inputs above ambient levels. To test the importance of CNU and recycled N to tree nutrition, we conducted a mesocosm experiment, applying 54 g N/¹⁵N ha^?1 yr^?1 to Sitka spruce saplings. We compared tree and soil ¹⁵N recovery among treatments where enrichment was due to either (1) a ¹⁵N‐enriched litter layer, or mineral ¹⁵N additions to (2) the soil or (3) the canopy. We found that 60% of ¹⁵N applied to the canopy was recovered above ground (in needles, stem and branches) while only 21% of ¹⁵N applied to the soil was found in these pools. ¹⁵N recovery from litter was low and highly variable. ¹⁵N partitioning among biomass pools and age classes also differed among treatments, with twice as much ¹⁵N found in woody biomass when deposited on the canopy than soil. Stoichiometrically calculated N effect on C uptake from ¹⁵N applied to the soil, scaled to real‐world conditions, was 43 kg C kg N^?1, similar to manipulation studies. The effect from the canopy treatment was 114 kg C kg N^?1. Canopy treatments may be critical to accurately represent N deposition in the field and may address the discrepancy between manipulative and correlative studies.     

Does light spectral quality affect survival and regeneration of potato (Solanum tuberosum L.) shoot tips after cryopreservation?

Cryopreservation, the storage of germplasm at ultra-low temperature is the most reliable tool for long-term preservation of plant genetic resources. Cryopreservation techniques are widely applied but the effect of light spectra on plant recovery after cryopreservation is largely unknown. Therefore, we investigated the effect of different light spectral qualities on survival and regeneration of shoot tips of potato (Solanum tuberosum L.) cultivars Agrie Dzeltenie, Maret, Bintje, Désirée and Anti cryopreserved by the DMSO-droplet method. Prior to cryopreservation, the plants were stored under cool white fluorescent light (CW). Post-cryopreservation, the plants were allowed to regenerate under six different light qualities: CW, warm white light (HQI), blue LEDs (B), red LEDs (R), red with 10 % of blue (RB) and RBF - red with 10 % of blue with addition of 20 % of far-red LEDs. The light spectral quality had a significant effect on the survival and regeneration of potato shoot tips after cryopreservation. The combination of red light with 10 % of blue (RB) doubled the regeneration percentage of all cultivars, whereas red light (R) was not suitable for regeneration after cryopreservation. Specifically, the regeneration percentages were increased in RB compared to CW from 25.5 to 52.6 % for ‘Agrie Dzeltenie’, 25.0–43.6 % for ‘Maret’, 8.1–26.1 % for ‘Bintje’, 0.0–17.1 % for ‘Anti’ and 18.2–36.6 % for ‘Désirée‘. Therefore, the modification of light spectra during the recovery phase is a promising tool for increasing the regeneration of potato shoot tips after cryopreservation.     

Stimulation of (Na+K+)-ATPase of rat liver plasma membrane by amino acids

The effect of twelve l-amino acids on the activity of liver plasma membrane (Na⁺K⁺)-ATPase has been tested. Histidine and arginine significantly enhanced the activity. The activtion by histidine showed saturation kinetics with an apparent K_a of about 8 mM, and was evident over a wide range of Na⁺ concentrations. The same amino acid did not significantly affect the Mg²⁺-dependent ATPase activity.     

Protective effects of allopurinol against acute liver damage and cirrhosis induced by carbon tetrachloride: Modulation of NF-κB, cytokine production and oxidative stress

Background

The aim of this work was to evaluate the hepatoprotective ability of allopurinol to prevent the liver injury induced by carbon tetrachloride (CCl₄).

Methods

Acute liver damage was induced with CCl₄ (4 g/kg, by gavage); allopurinol (50 mg/kg, by gavage) was given 1 h before and 1 h after CCl₄ intoxication and two daily doses for the previous three days. Cirrhosis was established by CCl₄ administration (0.4 g/kg, i. p. three times a week, eight weeks); allopurinol was administered (100 mg/kg, by gavage, daily) during the long-term of CCl₄ treatment. Alanine aminotransferase (ALT), γ-glutamyl transpeptidase (γ-GTP), xanthine oxidase (XO), lipid peroxidation, reduced and oxidized glutathione (GSH, GSSG, respectively), hydroxyproline and histopathologycal analysis were performed. Nuclear factor-κB (NF-κB), pro-inflammatory and anti-inflammatory cytokines, transforming growth factor-β (TGF-β) and metalloproteinase-13 (MMP-13) were analyzed by Western blots.

Results

Acute injury increased ALT and γ-GTP activities, additionally enhanced NF-κB nuclear translocation and cytokines production such as tumor necrosis factor-α, interleukine-1β, and interleukine-6. Allopurinol partially prevented these effects, while increased interleukine-10. Acute and chronic CCl₄ treatments altered the levels of XO activity, lipid peroxidation, and GSH/GSSG ratio, while these remained within normal range with allopurinol administration. Necrosis, fibrosis and TGF-β production induced in chronic injury were partially prevented by allopurinol, interestingly, this drug induced MMP-13 activity.

Conclusions

Allopurinol possesses antioxidant, anti-inflammatory and antifibrotic properties, probably by its capacity to reduce NF-κB nuclear translocation and TGF-β expression, as well as to induce MMP-13.General significanceAllopurinol might be effective treatment of liver diseases.     

Carnitine palmitoyltransferase activities (EC 2.3.1.–) of rat liver mitochondria

1. A continuously recording and sensitive fluorimetric assay is described for carnitine palmitoyltransferase. This assay has been applied to whole or disintegrated mitochondria and to soluble protein fractions. 2. When rat liver mitochondria had been disintegrated by ultrasound, the specific activity of carnitine palmitoyltransferase was 15-20m-units/mg of protein. Only one-fifth of this activity was assayable (with added substrates) before mitochondrial disintegration. 3. It is concluded that there are two carnitine palmitoyltransferase activities in rat liver mitochondria, of which one (type I) is relatively superficial in location and catalyses an acyl-group transfer between added CoA and carnitine, whereas the other (type II) is less superficial and catalyses an acyl-group transfer in unbroken mitochondria between added carnitine and intramitochondrial CoA. The existence of two distinct carnitine palmitoyltransferases was predicted by Fritz & Yue (1963). 4. In unbroken mitochondria, type I transferase is accessible to the inhibitor 2-bromostearoyl-CoA whereas the type II transferase is inaccessible. 5. A major part of the total carnitine palmitoyltransferase activity of rat liver mitochondria is membrane-bound and of type II. 6. These observations, when considered in conjunction with the penetration of mitochondria by CoASH or carnitine, indicate that the type II transferase is attached to the inner mitochondrial membrane.     

The proton-translocating nicotinamide–dinucleotide (phosphate) transhydrogenase of rat liver mitochondria

1. The NAD(P) transhydrogenase activity of the soluble fraction of sonicated rat liver mitochondrial preparations was greater than the NAD-linked isocitrate dehydrogenase activity, and the NAD-linked and NADP-linked isocitrate dehydrogenase activities were not additive. The NAD-linked isocitrate dehydrogenase activity was destroyed by an endogenous autolytic system or by added nucleotide pyrophosphatase, and was restored by a catalytic amount of NADP. 2. We concluded that the isocitrate dehydrogenase of rat liver mitochondria was exclusively NADP-specific, and that the oxoglutarate/isocitrate couple could therefore be used unequivocally as redox reactant for NADP in experiments designed to operate only the NAD(P) transhydrogenase (or loop 0) segment of the respiratory chain in intact mitochondria. 3. During oxidation of isocitrate by acetoacetate in intact, anaerobic, mitochondria via the rhein-sensitive, but rotenone- and arsenite-insensitive, NAD(P) transhydrogenase, measurements of the rates of carbonyl cyanide p-trifluoromethoxyphenylhydrazone-sensitive and carbonyl cyanide p-trifluoromethoxyphenylhydrazone-insensitive pH change in the presence of various oxoglutarate/isocitrate concentration ratios gave an -->H(+)/2e(-) quotient of 1.94+/-0.12 for outward proton translocation by the NAD(P) transhydrogenase. 4. Measurements with a K(+)-sensitive electrode confirmed that the electrogenicity of the NAD(P) transhydrogenase reaction corresponded to the translocation of one positive charge per acid equivalent. 5. Sluggish reversal of the NAD(P) transhydrogenase reaction resulted in a significant inward proton translocation. 6. The possibility that isocitrate might normally be oxidized via loop 0 at a redox potential of -450mV, or even more negative, is discussed, and implies that a P/O quotient of 4 for isocitrate oxidation might be expected.     

Toll like receptor 2 knock-out attenuates carbon tetrachloride (CCl4)-induced liver fibrosis by downregulating MAPK and NF-κB signaling pathways

Innate immune signaling associated with Toll-like receptors (TLRs) is a key pathway involved in the progression of liver fibrosis. In this study, we reported that TLR2 is required for hepatic fibrogenesis induced by carbon tetrachloride (CCl₄). After CCl₄ treatment, TLR2^−/− mice had reduced liver enzyme levels, diminished collagen deposition, decreased inflammatory infiltration and impaired activation of hepatic stellate cells (HSCs) than wild type (WT) mice. Furthermore, after CCl₄ treatment, TLR2^−/− mice demonstrated downregulated expression of profibrotic and proinflammatory genes and impaired mitogen-activated protein kinases (MAPK) and nuclear factor kappa B (NF-κB) activation than WT mice. Collectively, our data indicate that TLR2 deficiency protects against CCl₄-induced liver fibrosis.     

Dehydrogenation of androsterone by purified 3α-hydroxy steroid-dependent nicotinamide–adenine dinucleotide (phosphate)-transhydrogenating enzyme of rat liver

1. An enzyme from rat liver, catalysing 3alpha-hydroxy steroid-dependent NAD(P) transhydrogenation and NAD-linked and NADP-linked dehydrogenation of 3alpha-hydroxy steroids, has been purified 100-fold by chromatography on DEAE-cellulose and calcium phosphate gel. 2. No separation of these activities into different protein fractions has been achieved. 3. The properties of the enzyme in catalysing NAD-linked and NADP-linked dehydrogenation have been compared, with androsterone as substrate. Differences were found in pH optima, affinity for coenzyme and steroid, equilibrium constants and effects of salts. 4. NAD-linked dehydrogenation is inhibited by NADPH(2) but is protected from this inhibition by chloride, which alone is itself an inhibitor. 5. The relevance of these findings to the problem of the number of enzymes involved in catalysis of 3alpha-hydroxy steroid-dependent transhydrogenation is discussed.     

Augmenter of liver regeneration (ALR) gene therapy attenuates CCl₄-induced liver injury and fibrosis in rats

Liver fibrosis represents a process of healing and scarring in response to chronic liver injury. Augmenter of liver regeneration (ALR) has been shown to protect hepatocytes from various toxins. The aim of this study was to investigate the effects of ALR gene therapy on liver injury and fibrosis induced by CCl(4) in rats and further explore the underlying mechanisms. Human ALR expression plasmid was delivered via the tail vein. ALR gene therapy might protect the liver from CCl(4)-induced injury and fibrogenesis by attenuating the mitochondrial dysfunction, suppressing oxidative stress, and inhibiting activation of HSCs. This report demonstrated that ALR gene therapy protected against the ATP loss, increased the activity of ATPase, decreased intrahepatic reactive oxygen species level, and down-regulated transforming growth factor-β1, platelet-derived growth factor-BB, and α-smooth muscle actin expression. Following gene transfer liver function tests were significantly improved. In brief, ALR gene therapy might be an effective therapeutic reagent for liver fibrosis with potential clinical applications.     

The oxidation of N,N′-bis(4-aminophenyl)-N,N′-dimethylethylenediamine (BED) by rat liver

We have examined the oxidation of N,N′-bis(4-aminophenyl)-N,N′-dimethylethylenediamine (BED) by tissue homogenates and fractions of liver homogenates. We find that this agent both gives osmiophilic deposits in tissue blocks and readily increases the uptake of oxygen by hepatic homogenates. The highest activity was in the mitochondrial and, next, in the microsomal fractions. Kinetic evidence indicates that the former represents two enzymatic activities while the latter is only a single site. The activity was greatest in the outer membrane of the mitochondria, in agreement with electron micrographic studies and in the rough microsomal fraction. Further, it was very sensitive to both formaldehyde and detergents. The activity was not well associated with either monamine oxidase (benzylamine substrate) or xanthine oxidase activities. Activity was observed in a large number of tissues.     

In vitro induction of tumor necrosis factor-α by ochratoxin A (OTA) from rat liver: role of Kupffer cells

Tumor necrosis factor-α (TNF-α) is released from blood-free perfused rat liver by the fungal metabolite ochratoxin A. Here we have identified Kupffer cells as the sole source of OTA-mediated cytokine release. If single cell preparation of Kupffer cells, hepatocytes, or sinusoidal endothelial cells were prepared from rat livers, only Kupffer cells released TNF-α upon incubation with 2.5 μmol/l OTA. OTA failed to induce TNF-α release in the blood-free perfused isolated rat liver when Kupffer cells were blockedin vitro by 15 μmol/l gadolinium chloride. When rats were pretreatedin vivo with the Kupffer cell depleting clodronate liposomes, OTA-mediated TNF-α release was abrogated in the isolated perfused liver model.     

Specific inhibition of pyruvate transport in rat liver mitochondria and human erythrocytes by α-cyano-4-hydroxycinnamate (Short Communication)

alpha-Cyano-4-hydroxycinnamate greatly inhibits the transport of pyruvate but not that of acetate or butyrate in liver mitochondria and erythrocytes. In the latter, lactate uptake is also inhibited. It is concluded that a specific carrier is involved in membrane transport of pyruvate and that the plasma-membrane carrier may also be involved in lactate transport.     

Shoot regeneration and cryopreservation of shoot tips of apple (Malus) by encapsulation–dehydration

Here, we report an efficient and widely applicable method for cryopreservation of Malus shoot tips by encapsulation–dehydration using adventitious shoots. Shoots were induced from leaf segments cultured on a shoot induction medium containing 2–3 mg L^?1 thidiazuron, depending on genotype, and 0.5 mg L^?1 indole-3-butyric acid. Shoot tips (3 mm in length) containing six leaf primordia excised from 11-wk-old adventitious shoots were encapsulated and precultured with 0.5 M sucrose for 5 d, followed by air-drying for 6 h prior to direct immersion in liquid nitrogen. With our protocol, we obtained a mean organogenesis rate of 100%, a mean of 4.5 adventitious shoots per explant (leaf segment), and a mean shoot recovery of 57.0% from cryopreserved shoot tips in four Malus species. Inter-simple sequence repeat (ISSR) analysis did not reveal any polymorphic bands in regenerants recovered from either leaf segments or cryopreserved shoot tips of ‘Gala’. To the best of our knowledge, this is the first report on cryopreservation of Malus shoot tips using adventitious shoots derived from leaf segments and is the most widely applicable protocol so far reported for cryopreservation of Malus. Establishment of this protocol provides an alternative means for cryopreservation of Malus.     

Do hermaphrodites of gynodioecious Phacelia linearis (Hydrophyllaceae) trade off seed production to attract pollinators?

Sexual selection theory predicts that hermaphroditic plants might trade off seed production to attract pollinators. This paper reports a test of this prediction in gynodioecious Phacelia linearis (Hydrophyllaceae), a species in which attractiveness to pollinators increases with corolla diameter. The relationship between corolla diameter and seed production was determined in three natural and three experimental populations. Phenotypic selection analysis was used, with lifetime seed production as a surrogate for fitness. Negative directional selection was expected on hermaphrodite corolla diameter. No directional selection (but possibly stabilizing selection) was expected on corolla diameter in females, which have smaller corollas than hermaphrodites. Shoot biomass and flowering time were included in selection analyses so that the effects of corolla size could be assessed independendy of these correlated characters. A parent-offspring study of the quantitative genetics of these characters also was performed. High seed production was associated with large shoot biomass (especially in natural populations) and early flowering (especially in experimental populations). The effect of hermaphrodite corolla size on seed production was not as predicted. Directional selection on hermaphrodite corolla diameter was generally positive. Directional selection on female corolla diameter was relatively weak and variable; stabilizing selection was found in one population. Most of the phenotypic selection on corolla diameter was caused by its (environmental) correlation with shoot biomass. Although these findings do not support the hypothesis that hermaphrodites trade off seed production to attract pollinators, a cautious interpretation is warranted. In one natural population, insect herbivory generated positive covariance between hermaphrodite corolla diameter and seed number, leading to a spurious estimate of positive directional selection for corolla diameter. Other, undetected sources of positive covariance between corolla size and seed production also might have obscured the expected tradeoff. Corolla diameter was found to be heritable, but shoot biomass and flowering time were not. The genetic correlation between hermaphrodite corolla diameter and female corolla diameter was positive and significant, but it was significantly less than 1, so corolla size could evolve independently in the two genders.     

Kinetics of oxygen and carbon monoxide binding to liver fluke (Dicrocoelium dendriticum) hemoglobin. An extreme case?

The kinetics of oxygen and carbon monoxide binding to the monomeric liver fluke (Dicrocoelium dendriticum) hemoglobin have been studied. The ligand association rates are approximately 1 X 10(8) and approximately 3 X 10(8) M-1 s-1, respectively, for CO and O2 and show no pH dependence. On the contrary the ligand dissociation rates decrease by lowering the pH below 7, the pK of the transition being around 5.5. These findings, together with spectroscopic properties of the protein, are discussed in relation to the fact that, in this hemoglobin, the distal histidine is replaced by a glycine.     

Effect of selenite and selenate on rat liver nuclear 3,5,3′-triiodothyronine (T3) receptor

The present study was undertaken to investigate the effects of selenite (Se^IV) or selenate (Se^VI) on nuclear T₃ receptors of rat liver. Selenite at 0.1 μM (p<0.01) inhibited the T₃ specific binding to rat liver nuclear receptors. The specific binding of the T₃ receptor was fully restored when even 1.0 μM selenite was separated from the T₃ receptor by gel filtration. No inhibitory effect of selenite (up to 100 μM) on the T₃ binding to nuclear receptor was found in the presence of 1.0 mM dithiothreitol. The rate of dissociation of the T₃-nuclear receptor complex was effectively increased by 0.1 μM selenite. Selenate up to 1 mM as well as sulfite or sulfate up to 0.1 mM did not exert an inhibitory effect on T₃ receptors. The results based on the in vitro experiments suggest that the selenium in the form of selenite may reversibly affect the T₃ binding on the receptor molecule.