Intestinal absorption of forsythoside A in different compositions of Shuang-Huang-Lian

Shuang-Huang-Lian (SHL), a traditional Chinese formula containing Lonicerae japonicae flos (LJF), Scutellariae radix (SR) and Forsythiae fructus (FF), is commonly used to treat acute upper respiratory tract infection, acute bronchitis and light pneumonia. Forsythoside A is one of the main active ingredients in Forsythiae fructus, a key herb in SHL. In the present study, effects of different compositions in SHL on the intestinal absorption of forsythoside A were investigated. The observations from in situ intestinal circulation model showed that A/%(h^− 1) of forsythoside A in FF + LSF, FF + SR and SHL were all reduced greatly compared with that in FF. However, in pharmacokinetics study, C_max and AUC_0 → 1440 of forsythoside A all increased and T_1/2 prolonged in SHL, FF + LJF and FF + SR compared with FF. The results indicated that the different compositions of SHL decreased absorption but increased bioavailability of forsythoside A, which may be related to its metabolism inhibited in intestine or liver.     

In vitro metabolism in Sprague-Dawley rat liver microsomes of forsythoside A in different compositions of Shuang-Huang-Lian

Shuang–Huang–Lian (SHL), a traditional Chinese formula containing Lonicerae japonicae flos (LJF), Scutellariae radix (SR) and Forsythiae fructus (FF), is commonly used to treat acute upper respiratory tract infection, acute bronchitis and light pneumonia. Forsythoside A is one of the main active ingredients in Forsythiae fructus, a key herb in SHL. In the present study, effects of different compositions in SHL on the in vitro metabolism in Sprague–Dawley rat liver microsomes of forsythoside A were investigated. The observations from Sprague–Dawley rat liver microsomes in the presence of β-NADPH or UDPGA that forsythoside A may be the substrates of CYP3A4, CYP2C9, CYP1A2, UGT1A6, UGT1A3, UGT1A1 and UGT1A9; Chlorogenic acid may be the substrates of CYP3A4, CYP2C9, CYP1A2, CYP2C19, UGT1A6, UGT1A3 and UGT1A1; Baicalin may be the substrates of CYP3A4, CYP2C19, CYP1A2, UGT1A9, UGT1A1 and UGT1A3; Baicalein may be the substrates of CYP3A4, CYP2E1 and UGT1A6. It was also found that the residue of forsythoside A in SHL, FF + LJF and FF + SR was greatly increased compared with that in FF in Sprague–Dawley rat liver microsomes in the presence of β-NADPH or UDPGA, which indicated that the metabolism of forsythoside A in SHL may be influenced by chlorogenic acid in LJF acting on the CYP3A4, CYP2C9, CYP1A2, UGT1A6, UGT1A3 and UGT1A1; baicalin in SR acting on the CYP3A4, CYP1A2, UGT1A9, UGT1A1 and UGT1A3; baicalein acting on the CYP3A4 and UGT1A6 respectively.     

Hypoglycemic effect of protopanaxadiol-type ginsenosides and compound K on Type 2 diabetes mice induced by high-fat diet combining with streptozotocin via suppression of hepatic gluconeogenesis

Compound K (CK) is a final intestinal metabolite of protopanaxadiol-type ginsenosides (PDG) from Panax ginseng. Although anti-diabetic activity of CK have been reported with genetic mouse models (db/db mice) in recent years, the therapeutic usefulness of CK and PDG in type 2 diabetes, a more prevalent form of diabetes, remains unclear. In the present investigation, we developed a mouse of non-insulin-dependent diabetes mellitus that closely simulated the metabolic abnormalities of the human disease. For this purpose, type 2 diabetes was induced in male ICR mice by combining of streptozotocin. The male ICR mice fed with HFD for 4 weeks received 100 mg/kg of STZ injected intraperitoneally. After 4 weeks, mice with fasting (12 h) blood glucose levels (FBG) above 7.8 mmol/L were divided into 3 groups (n = 12) and treated with vehicle (diabetes model, DM), 300 mg/kg/day of PDG and 30 mg/kg/day of CK for 4 weeks while continuing on the high-fat diet. Hypoglycemic effects of CK and PDG were consistently demonstrated by FBG levels, and insulin-sensitizing effects were seen during oral glucose tolerance testing (OGTT). Moreover, the mechanism of hypoglycemic effect in type 2 diabetic mice was examined. Gluconeogenic genes, Phosphoenolpyruvate carboxykinase (PEPCK) and Glucose-6-phosphatase (G6Pase), were decreased in two treatment groups with CK showing greater effects. These findings demonstrated the hypoglycemic and insulin-sensitizing capabilities of CK on type 2 diabetes induced by HFD/STZ via down-regulation of PEPCK and G6Pase expression in liver.     

HPLC-based activity profiling of Angelica pubescens roots for new positive GABAA receptor modulators in Xenopus oocytes

A petroleum ether extract of the traditional Chinese herbal drug Duhuo (roots of Angelica pubescens Maxim. f. biserrata Shan et Yuan), showed significant activity in a functional two-microelectrode voltage clamp assay with Xenopus oocytes which expressed recombinant γ-aminobutyric acid type A (GABA_A) receptors of the subtype α₁β₂γ_2S. HPLC-based activity profiling of the active extract revealed six compounds responsible for the GABA_A receptor modulating activity. They were identified by microprobe NMR and high resolution mass spectrometry as columbianetin acetate (1), imperatorin (3), cnidilin (4), osthol (5), and columbianedin (6). In concentration-dependent experiments, osthol and cnidilin showed the highest potentiation of the GABA induced chloride current (273.6% ± 39.4% and 204.5% ± 33.2%, respectively at 300 μM). Bisabolangelone (2) only showed minor activity at the GABA_A receptor. The example demonstrates that HPLC-based activity profiling is a simple and efficient method to rapidly identify GABA_A receptor modulators in a bioactive plant extract.     

Comparative pharmacokinetics of paclitaxel after oral administration of Taxus yunnanensis extract and pure paclitaxel to rats

Taxus yunnanensis (T. yunnanensis) is endemic to China and has been used in traditional medicine for the treatment of cancer, diabetic ailments and others. Paclitaxel is a representative antitumor compound in the Taxus species. The pharmacokinetic behavior of paclitaxel after oral administration of the crude extract of T. yunnanensis has not been investigated. This study attempts to compare the pharmacokinetics of paclitaxel after an oral administration of the crude extract of the twigs and leaves of T. yunnanensis and pure paclitaxel. A UPLC and a UPLC/MS/MS analysis method were developed for the determination of paclitaxel in T. yunnanensis extract and in the comparative pharmacokinetic study. Caco-2 cells were used to investigate the transport profile of paclitaxel in vitro. In the pharmacokinetic study, rats were randomly grouped and administered with T. yunnanensis extract or pure paclitaxel. The results showed that the AUC and C_max of paclitaxel in rats receiving the T. yunnanensis extract were significantly increased than those receiving the pure paclitaxel, and the in vitro Caco-2 cell monolayer transport study found that the coexisting constituents in the extract of T. yunnanensis could inhibit the efflux of paclitaxel. These findings suggested that the oral absorption and bioavailability of paclitaxel in T. yunnanensis extract were remarkably higher when compared with the pure paclitaxel, and the coexisting constituents in the T. yunnanensis extract might play an important role for the enhancement of the oral absorption and bioavailability of paclitaxel.     

Soil–atmosphere CO2, CH4 and N2O fluxes in boreal forestry-drained peatlands

Greenhouse gas emissions from managed peatlands are annually reported to the UNFCCC. For the estimation of greenhouse gas (GHG) balances on a country-wide basis, it is necessary to know how soil–atmosphere fluxes are associated with variables that are available for spatial upscaling. We measured momentary soil–atmosphere CO₂ (heterotrophic and total soil respiration), CH₄ and N₂O fluxes at 68 forestry-drained peatland sites in Finland over two growing seasons. We estimated annual CO₂ effluxes for the sites using site-specific temperature regressions and simulations in half-hourly time steps. Annual CH₄ and N₂O fluxes were interpolated from the measurements. We then tested how well climate and site variables derived from forest inventory results and weather statistics could be used to explain between-site variation in the annual fluxes. The estimated annual CO₂ effluxes ranged from 1165 to 4437 g m⁻² year⁻¹ (total soil respiration) and from 534 to 2455 g m⁻² year⁻¹ (heterotrophic soil respiration). Means of 95% confidence intervals were ±12% of total and ±22% of heterotrophic soil respiration. Estimated annual CO₂ efflux was strongly correlated with soil respiration at the reference temperature (10 °C) and with summer mean air temperature. Temperature sensitivity had little effect on the estimated annual fluxes. Models with tree stand stem volume, site type and summer mean air temperature as independent variables explained 56% of total and 57% of heterotrophic annual CO₂ effluxes. Adding summer mean water table depth to the models raised the explanatory power to 66% and 64% respectively. Most of the sites were small CH₄ sinks and N₂O sources. The interpolated annual CH₄ flux (range: −0.97 to 12.50 g m⁻² year⁻¹) was best explained by summer mean water table depth (r² = 64%) and rather weakly by tree stand stem volume (r² = 22%) and mire vegetation cover (r² = 15%). N₂O flux (range: −0.03 to 0.92 g m⁻² year⁻¹) was best explained by peat CN ratio (r² = 35%). Site type explained 13% of annual N₂O flux. We suggest that water table depth should be measured in national land-use inventories for improving the estimation of country-level GHG fluxes for peatlands.     

Swietenine: A potential oral hypoglycemic from Swietenia macrophylla seed

Swietenine, a tetranortriterpenoid, was isolated from the Swietenia macrophylla seeds. The in vivo hypoglycemic activity was evaluated against neonatal-streptozotocin induced type 2 diabetic rats. Oral administration of swietenine at 25 and 50 mg/kg body weight per day to diabetic rats was found to possess significant dose dependant hypoglycemic and hypolipidemic activity in type 2 diabetic rats.     

Ultra-performance liquid-chromatography with tandem mass spectrometry performing pharmacokinetic and biodistribution studies of croomine,neotuberostemonine and tuberostemonine alkaloids absorbed in the rat plasma after oral administration of Stemonae Radix

Stemonae Radix (Stemona tuberosa Lour, Bai Bu) is an important traditional Chinese medicinal (TCM) plant known for its antitussive activity. Croomine, neotuberostemonine and tuberostemonine alkaloids of Stemonae Radix are major components responsible for antitussive action. In this work, plasma pharmacokinetic and biodistribution characteristics of the three alkaloids after oral administration of Stemonae Radix are investigated using a rapid and sensitive UPLC-Q-TOF–HDMS method. Mass spectrometry (MS) was performed on a Waters Micromass high-definition technology with an electrospray ionization source in positive ion mode, with excellent MS mass accuracy and enhanced MS data acquisition. Separation of main alkaloids was achieved on a Waters BEH C₁₈ column by linear gradient elution. Data were analyzed and estimated by compartmental methods and pharmacokinetic parameters calculated using WinNonlin Professional version 5.1. It was found that croomine, neotuberostemonine and tuberostemonine had faster absorbed into the bloodstream, maintain the high plasma concentration, and pose a large AUC value. The biodistribution of neotuberostemonine and tuberostemonine showed that the higher levels were in liver, and lung. Croomine was discovered in brain and showed that it could cross the blood–brain barrier, indicating that croomine plays an antitussive effect as acting on the central nervous system. Neotuberostemonine and tuberostemonine were not discovered in brain, demonstrating that they play an antitussive effect as peripheral antitussive. This work suggests that the pharmacokinetics and biodistribution based-UPLC-Q-TOF–HDMS can provide a reliable tools for screening bioactive components contributing to pharmacological effects of medicinal herbs.     

Pharmacokinetic study of luteolin,apigenin, chrysoeriol and diosmetin after oral administration of Flos Chrysanthemi extract in rats

Flos Chrysanthemi (the flower of Chrysanthemum morifolium Ramat.) is widely used in China as a food and traditional Chinese medicine for many diseases. Luteolin and apigenin are two main bioactive components in Flos Chrysanthemi, and chrysoeriol and diosmetin are two methylated metabolites of luteolin in vivo by cathechol-O-methyltransferase (COMT). However, there was lack of pharmacokinetic information of chrysoeriol and diosmetin after oral administration of Flos Chrysanthemi extract (FCE). The present study aimed to develop an HPLC-UV method for simultaneous determination of rat plasma concentration of luteolin, apigenin, chrysoeriol and diosmetin and utilize it in pharmacokinetic study of the four compounds after orally giving FCE to rats. The method was successfully validated and applied to the pharmacokinetic study when oral administration of FCE to rats with or without co-giving a COMT inhibitor, entacapone. Chrysoeriol and diosmetin were detected in rat plasma after oral administration of FCE and their concentrations were significantly decreased after co-giving entacapone. Furthermore, AUC of luteolin was significantly increased by entacapone, while that of chrysoeriol was decreased by entacapone, which revealed COMT might play an important role in the disposition of luteolin in rats after dosing of FCE. In conclusion, a sensitive, accurate and reproducible HPLC-UV method for simultaneous determination of luteolin, apigenin, chrysoeriol and diosmetin in rat plasma were developed, pharmacokinetics of chrysoeriol and diosmetin combined with luteolin and apigenin were characterized after oral administration of FCE to rats, which gave us more information on pharmacokinetics and potential pharmacological effects of FCE in vivo.     

Reactive oxygen species scavenging activities and inhibition on DNA oxidative damage of dimeric compounds from the oxidation of (−)-epigallocatechin-3-O-gallate

The dimeric catechins dehydrotheasinensin A (2) and theacitrin C (3) were prepared from the oxidation of (−)-epigallocatechin-3-O-gallate (EGCG, 1), and their antioxidant activity was investigated using a chemiluminescence (CL) method in vitro. Both compounds showed significant inhibitory effects on reactive oxygen species (O₂⁻, H₂O₂ and •OH) and DNA oxidative damage, with 2 being more potent than 3 and EGCG itself.     

Hypoglicemic effect of Leandra lacunosa in normal and alloxan-induced diabetic rats

Leandra lacunosa, popularly known as "erva-do-jabuti", is used in Brazilian folkloric medicine for the treatment of diabetes mellitus. Based on this traditional indication, the aim of this work was to evaluate the hypoglycemic activity of the hydroalcoholic extract of L. lacunosa aerial parts (LLH) in normal and alloxan-induced diabetic rats. Chromatographic fractionation of LLH was also carried out by several techniques, affording isolation of the following major compounds: ursolic acid (1), kaempferol (2), luteolin (3), and quercetin (4). The oral administration of LLH (500 mg/kg) in normal rats caused a significant reduction of 24.7% (P<0.05) in the blood glucose levels after 2 h of treatment, while the administration of chlorpropamide (20 mg/kg, p.o.) led to a reduction of 40.2% (P<0.01). After oral administration of glucose (10 g/kg, p.o.), LLH (500 mg/kg, p.o.) significantly inhibited the increase in blood glucose levels compared with the negative control group. The oral treatment with LLH (500 mg/kg) in alloxan-induced diabetic rats significantly reduced the blood glucose levels in 47.8% after 4 h of treatment, while chlorpropamide resulted in a significant reduction of 71.7% in the 4th hour. Our results showed that LLH, displays hypoglycemic activity, which may be related to the effect of the major compounds identified in the crude extract. This study seems to provide biological evidence for the folkloric use of L. lacunosa in the treatment of diabetes mellitus.     

Pharmacological mechanisms of black cohosh in Sprague-Dawley rats

Background

Studies indicate that extracts and purified components from black cohosh inhibit the growth of human breast cancer cells, but the molecular targets and signaling pathways have not yet been defined.

Purpose

This study examines the pharmacological mechanisms and toxicological effects in the short term of the herb black cohosh on female Sprague–Dawley rats.

Materials and methods

To assess effects on gene activity and lipid content, we treated female Sprague–Dawley rats with an extract of black cohosh enriched in triterpene glycosides (27%) at 35.7 or 0 mg/kg. Four animals for each group were sacrificed at 1, 6 and 24 h after treatment; liver tissue and serum samples were obtained for gene expression and lipid analysis.

Results

Microarray analysis of rat liver tissue indicated that black cohosh markedly downregulated mitochondrial oxidative phosphorylation genes. Phospholipid biosynthesis and remodeling, PI3-Kinase and sphingosine signaling were upregulated, driven largely by an upregulation of several isoforms of phospholipase C. Hierarchical clustering indicated that black cohosh clustered with antiproliferative compounds, specifically tubulin binding vinca alkaloids and DNA alkylators. In support of this, black cohosh repressed the expression of cyclin D1 and ID3, and inhibited the proliferation of HepG2, p53 positive, liver cancer cells. Black cohosh reduced the level of free fatty acids at 6 and 24 h and triglycerides at 6 h in the serum, but increased the free fatty acid and triglyceride content of the treated livers at 24 h.

Conclusion

Our results suggest that black cohosh warrants further study for breast cancer prevention and therapy.     

Bilberry extract protect restraint stress-induced liver damage through attenuating mitochondrial dysfunction

The aim of the present study is to investigate the protective effect of bilberry extract on liver damage in restraint stressed mice. A remarkable increase in alanine aminotransferase (ALT) and reactive oxygen species (ROS) levels was observed in stressed mice. Treatment with bilberry extract restored ALT and ROS to normal levels, and enhanced mitochondrial complex II activity that was lowered in restraint stressed mice. The mitochondrial electron transfer chain (ETC)-related gene expression was measured by RT-PCR, and a significant up-regulation of complex II mRNA was observed for SDHA, B, C and D mRNA in bilberry extract-treated group compared with that in stressed group. Bilberry extract administration also profoundly elevated the Na⁺-K⁺-ATPase activity and mitochondria membrane potential (?Ψ_m), which was reduced in the stressed group. Bilberry extract exhibited protective effect by scavenging free radicals and attenuating mitochondrial dysfunction in the liver of restraint stressed mice. It may be used as a promising therapeutic agent in preventing and delaying the life-related disease.     

Anti-diabetic effects of emodin involved in the activation of PPARγ on high-fat diet-fed and low dose of streptozotocin-induced diabetic mice

Rheum palmatum Linn has been widely applied in the clinical treatment of diabetes mellitus. It has been found that emodin as the major bioactive component of R. palmatum L exhibits the competency to activate peroxisomal proliferator-activated receptor-γ (PPARγ) in vitro. So the aim of this study was to evaluate the anti-diabetic effects of emodin through the activation of PPARγ on high-fat diet-fed and low dose of streptozotocin (STZ)-induced diabetic mice. The diabetic mice were intraperitoneally injected with emodin for three weeks. No changes of food consumption and the body weight in emodin-treated mice were monitored daily during the entire experiment. At the end of experiment, the levels of blood glucose, triglyceride and total cholesterol in serum were significantly decreased after emodin treatment. However, serum high-density lipoprotein cholesterol (HDLc) concentration was significantly elevated. The glucose tolerance and insulin sensitivity in emodin-treated group were significantly improved. Furthermore, the results of quantitative RT-PCR analysis showed that emodin significantly elevated the mRNA expression level of PPARγ and regulated the mRNA expressions of LPL, FAT/CD36, resistin and FABPs (ap2) in liver and adipocyte tissues. No effects on the mRNA expressions of PPARα and PPARα-target genes were observed. Taken together, the results suggested that the activation of PPARγ and the modulation of metabolism-related genes were likely involved in the anti-diabetic effects of emodin.     

Inhibition of glucose intestinal absorption by kaempferol 3-O-α-rhamnoside purified from Bauhinia megalandra leaves

Glucose intestinal absorption (GIA) is one of the factors that increase glycemia. Its reduction could be an important factor in decreasing hyperglycemia in diabetic patients. It has been shown that the aqueous extract of Bauhinia megalandra leaves inhibits GIA. In the present study we identified a compound present in the extract of B. megalandra responsible for the biological effect. The methanol extract of B. megalandra leaves was fractionated using different solvents, and high-speed counter-current chromatography yielding two pure compounds identified by ¹H NMR and ¹³C NMR as kaempferol 3-O-α-rhamnoside and quercetin 3-O-α-rhamnoside. The first one increased the K_M without changes in the V_MAX of GIA. In addition it exerted an additive inhibitory effect, on GIA, when combined with phlorizin. We suggest that kaempferol 3-O-α-rhamnoside is a competitive inhibitor of intestinal SGLT1 cotransporter.     

Anti-hyperglycemic effect of Potentilla discolor decoction on obese-diabetic (Ob-db) mice and its chemical composition

Potentilla discolor is used as an ethnomedicine in treatments of diabetes mellitus in China for years. In the present study, the anti-hyperglycemic effects of a clinical active extract (decoction) from P. discolor were investigated in Ob-db mice. Four week's treatment of P. discolor decoction ameliorated the development of hyperlipidemia, lipid peroxidation and hyperglycemia associated with hyperphagia and polydypsia in Ob-db mice. P. discolor significantly attenuated the increase of blood glucose and cholesterol levels in Ob-db mice. These findings clearly provided evidences regarding the anti-hyperglycemic potentials of P. discolor decoction. High-resolution liquid chromatography-mass spectrometry/mass spectrometry (HR-LC-MS/MS) was used to analyze the phytochemicals in P. discolor decoction. In an comprehensive analysis of phytochemicals in P. discolor, thirty‐five components were identified or characterized in P. discolor decoction and only sixteen of them have been reported in P. discolor previously. There are five major components identified in P. discolor decoction. One of the major components is a flavonoid sulfate, and this is the first evidence for the presences of sulfated flavonoid in P. discolor. Sulfated flavonoids have been reported to improve the complications of diabetes mellitus by inhibition of the aldose reductase in both experimental animals and clinical trials. Therefore, the sulfated flavonoid in P. discolor decoction may in part contribute to the anti-hyperglycemic effect of P. discolor.     

Potential insulin secretagogue effects of isovitexin and swertisin isolated from Wilbrandia ebracteata roots in non-diabetic rats

The antihyperglycemic effect and mechanism of action of extracts, fractions and compounds from Wilbrandia ebracteata was studied. The crude extract reduced the glycemia, increased glycogen content and serum insulin in hyperglycemic rats. Also, a significant effect was observed with the n-butanol and metanol subfraction. However, the antihyperglycemic effect of the n-butanol fraction was not observed in diabetic rats. The C-glycosylflavones isovitexin and swertisin showed a strong antihyperglycemic action compared with the extracts and fractions. These results show that the extracts, fractions, and isolated C-glycosylflavones have an antihyperglycemic action that was reinforced by the stimulation on in vivo insulin secretion.     

Cathartic effect of the leaf extract of Vernonia amygdalina

A methanol extract of Vernonia amygdalina leaves was evaluated for cathartic effect. The extract exhibited a significant promotion of intestinal motility on charcoal meal test in mice. Frequency of defecation and of faeces were markedly increased following administration of the extract which also promoted gastric emptying in rats. Preliminary studies on the isolated rat fundus strip showed a contractile effect, which was blocked by atropine.     

Pharmacological studies on the leaf of Psidium guajava

The methanol extract of the leaves of Psidium guajava was found to inhibit paw oedema induced by carrageenan in rats and pain induced by acetic acid in mice, and exhibited an antipyretic effect. Oral administration of the extract reduced intestinal transit time and prevented castor oil-induced diarrhoea in mice. A CNS depressant activity was exhibited by the extract by potentiating the phenobarbitone sleeping time in mice.     

Effects of irrigation on water use and water use efficiency in two fast growing Eucalyptus plantations

Eucalyptus plantations occupy almost 20 million ha worldwide and exceed 3.7 million ha in Brazil alone. Improved genetics and silviculture have led to as much as a three-fold increase in productivity in Eucalyptus plantations in Brazil and the large land area occupied by these highly productive ecosystems raises concern over their effect on local water supplies. As part of the Brazil Potential Productivity Project, we measured water use of Eucalyptus grandis × urophylla clones in rainfed and irrigated stands in two plantations differing in productivity. The Aracruz (lower productivity) site is located in the state of Espirito Santo and the Veracel (higher productivity) site in Bahia state. At each plantation, we measured stand water use using homemade sap flow sensors and a calibration curve using the clones and probes we utilized in the study. We also quantified changes in growth, leaf area and water use efficiency (the amount of wood produced per unit of water transpired). Measurements were conducted for 1 year during 2005 at Aracruz and from August through December 2005 at Veracel. Transpiration at both sites was high compared to other studies but annual estimates at Aracruz for the rainfed treatment compared well with a process model calibrated for the Aracruz site (within 10%). Annual water use at Aracruz was 1394 mm in rainfed treatments versus 1779 mm in irrigated treatments and accounted for approximately 67% and 58% of annual precipitation and irrigation inputs respectively. Increased water use in the irrigated stands at Aracruz was associated with higher sapwood area, leaf area index and transpiration per unit leaf area but there was no difference in the response of canopy conductance with air saturation deficit between treatments. Water use efficiency at the Aracruz site was also not influenced by irrigation and was similar to the rainfed treatment. During the period of overlapping measurements, the response to irrigation treatments at the more productive Veracel site was similar to Aracruz. Stand water use at the Veracel site totaled 975 mm and 1102 mm in rainfed and irrigated treatments during the 5-month measurement period respectively. Irrigated stands at Veracel also had higher leaf area with no difference in the response of canopy conductance with air saturation deficit between treatments. Water use efficiency was also unaffected by irrigation at Veracel. Results from this and other studies suggest that improved resource availability does not negatively impact water use efficiency but increased productivity of these plantations is associated with higher water use and should be given consideration during plantation management decision making processes aimed at increasing productivity.