Cytotoxicity evaluation of ketoprofen found in pharmaceutical wastewater on HEK 293 cell growth and metabolism.

Inefficient ketoprofen removal from pharmaceutical wastewater may negatively impact the ecosystem and cause detrimental risks to human health. This study was conducted to determine the cytotoxicity effects of ketoprofen on HEK 293 cell growth and metabolism, including cyclooxygenase-1 (COX-1) expression, at environmentally relevant concentrations. The cytotoxic effects were evaluated through the trypan blue test, DNS assay, MTT assay, and the expression ratio of the COX-1 gene. The results of this study show insignificant (p > 0.05) cytotoxic effects of ketoprofen on cell viability and cell metabolism. However, high glucose consumption rates among the treated cells cause an imitation of the Warburg effect, which is likely linked to the development of cancer cells. Apart from that, the upregulation of COX-1 expression among the treated cells indicates remote possibility of inflammation. Although no significant cytotoxic effects of ketoprofen were detected throughout this study, the effects of prolonged exposure of residual ketoprofen need to be evaluated in the future.

Assessment of heavy metals by ICP-OES and their impact on insulin stimulating hormone and carbohydrate metabolizing enzymes.

Arsenic (As) and cadmium (Cd) have recently emerged as major health concerns owing to their strong association with diabetes mellitus (DM). We aimed to investigate the heavy metals exposure towards incidence of DM at various enzymatic and hormonal levels. Additionally, association of As and Cd with Zinc (Zn, essential metal) was also evaluated. Spot urine samples were collected to assess As, Cd and Zn through ICP-OES. Serum was analyzed by assay method for fasting blood glucose, liver and renal function biomarkers. ELISA was performed to investigate the impact of heavy metals on HbA1c, α-amylase, DPP-IV, IGF-1, leptin, GSH, MDA, SOD, HDL, FFA, TG and interleukin (IL)-6. Association of heavy metals with DM was measured by odds ratio (OR) and level of significance was assessed by Chi-squared test. Unpaired student's t-test was used to compare DM-associated risk factors in heavy metals-exposed and unexposed participants. As and Cd were detectable in 75.4% and 83% participants with mean concentration of 75.5 ppb and 54.5 ppb, respectively. For As exposure, OR in the third quartile was maximum ie 1.34 (95% CI, 0.80 to 2.23), however the result was not statistically significant (P > .05). For Cd exposure, OR in the fourth quartile was considerably high, 1.62 (95% CI, 1.00 to 2.61), with a significant probability value (P < .05). Urinary Cd was negatively associated with Zn. As and Cd exposure increases the incidence of DM in the general population. Impaired hormonal and enzymatic levels in diabetic and non-diabetic exposed participants reflect the multiple organ damage by heavy metal exposure.

USING OF CELL LINE HepG2 FOR ASSESSMENT OF TOXIC AND METABOLIC EFECTS OF ANTIPSYCHOTIC DRUGS.

In order to study in vitro the toxic and metabolic effects of antipsychotic drugs (AP) on the cells of hepatic origin we used human hepatoblastoma cell line HepG2. We cultured HepG2 cells in the presence of two AP of the first and second generations (haloperidol and olanzapine, respectively) adding them to the culture medium in concentrations that may at the therapeutic use of AP take place in liver and other tissues of a high lipid content. In the process of cultivation, we detected several products of carbohydrate and lipid metabolism, measured activity of four hepatocellular enzymes in the culture medium, and estimated cell viability/proliferation in the MTS-test. We observed that both AP performed a toxic effect on HepG2 cells, the effect was manifested by a decrease in cell viability/proliferation and an increase in alkaline phosphatase activity in the culture medium. The toxic effect of olanzapine was less pronounced in comparison to haloperidol. According to the data from literature, AP upregulate the expression of lipogenesis genes in the cells of central nervous system, adipose tissue and liver, that might lead to hyperlipidemia. However, we observe in our experiments no increase in the levels of total cholesterol, of cholesterol in lipoproteins of high and low density, of triglycerides in the culture medium containing haloperidol or olanzapine. That observation may have been due to the fact that both AP, which are cationic amphiphiles, are capable to inhibit intracellular traffic of lipids. We also found no effects of haloperidol and olanzapine on the activity of aspartate aminotransferase and gamma-glutamyltransferase, while both AP did reduce the alanine aminotransferase activity. Our work proves that HepG2 cells can be helpful as an in vitro model to obtain new data on metabolic effects of drugs on the cells of hepatic origin and to assess the risk of a drug hepatotoxicity in preclinical studies.

Applications in environmental risk assessment of biochemical analysis on the Indian fresh water fish, Labeo rohita exposed to monocrotophos pesticide.

Pesticides are widely used in modern agriculture to aid in the production of high quality food. However, some pesticides have the potential to cause serious health and environmental damage. Repeated exposure to sub-lethal doses of pesticides can cause physiological and behavioral changes in fish that reduce populations such as abandonment of nests and broods, decreased immunity to disease and increased failure to avoid predators. Monocrotophos is one of the organophosphorus pesticide used in this study. The median lethal concentration (LC50) of Monocrotophos to fish L. rohita for 96h was found to be 45.1ppm. In sublethal concentration (1/10th of LC50 96h value, 4.51ppm) fishes were exposed for 24, 48, 72, 96h and 10, 20 and 30days. Organs of fishes were sacrificed and tested for biochemical analysis. A significant decrease in protein, carbohydrate and lipids were observed throughout the study period when compared to the control. It is essential for assessing the ecological risk of these pesticides.

Dapagliflozin, a Sodium-Glucose Co-Transporter 2 Inhibitor, Acutely Reduces Energy Expenditure in BAT via Neural Signals in Mice.

Selective sodium glucose cotransporter-2 inhibitor (SGLT2i) treatment promotes urinary glucose excretion, thereby reducing blood glucose as well as body weight. However, only limited body weight reductions are achieved with SGLT2i treatment. Hyperphagia is reportedly one of the causes of this limited weight loss. However, the effects of SGLT2i treatment on systemic energy expenditure have not been fully elucidated. Herein, we investigated the acute effects of dapagliflozin, a SGLT2i, on systemic energy expenditure in mice. Eighteen hours after dapagliflozin treatment oxygen consumption and brown adipose tissue (BAT) expression of ucp1, a thermogenesis-related gene, were significantly decreased as compared to those after vehicle treatment. In addition, dapagliflozin significantly suppressed norepinephrine (NE) turnover in BAT and c-fos expression in the rostral raphe pallidus nucleus (rRPa) which contains the sympathetic premotor neurons responsible for thermogenesis. These findings indicate that the dapagliflozin-mediated acute decrease in energy expenditure involves a reduction in BAT thermogenesis via decreased sympathetic nerve activity from the rRPa. Furthermore, common hepatic branch vagotomy abolished the reductions in ucp1 expression and NE contents in BAT and c-fos expression in the rRPa. In addition, alterations in hepatic carbohydrate metabolism, such as decreases in glycogen contents and upregulation of phosphoenolpyruvate carboxykinase, manifested prior to the suppression of BAT thermogenesis, e.g. 6 hours after dapagliflozin treatment. Collectively, these results suggest that SGLT2i treatment acutely suppresses energy expenditure in BAT via regulation of an inter-organ neural network consisting of the common hepatic vagal branch and sympathetic nerves.

Impact of zinc supplementation on the improved fructose/xylose utilization and butanol production during acetone-butanol-ethanol fermentation.

Lignocellulosic biomass and dedicated energy crops such as Jerusalem artichoke are promising alternatives for biobutanol production by solventogenic clostridia. However, fermentable sugars such as fructose or xylose released from the hydrolysis of these feedstocks were subjected to the incomplete utilization by the strains, leading to relatively low butanol production and productivity. When 0.001 g/L ZnSO4·7H2O was supplemented into the medium containing fructose as sole carbon source, 12.8 g/L of butanol was achieved with butanol productivity of 0.089 g/L/h compared to only 4.5 g/L of butanol produced with butanol productivity of 0.028 g/L/h in the control without zinc supplementation. Micronutrient zinc also led to the improved butanol production up to 8.3 g/L derived from 45.2 g/L xylose as sole carbon source with increasing butanol productivity by 31.7%. Moreover, the decreased acids production was observed under the zinc supplementation condition, resulting in the increased butanol yields of 0.202 g/g-fructose and 0.184 g/g-xylose, respectively. Similar improvements were also observed with increasing butanol production by 130.2 % and 8.5 %, butanol productivity by 203.4% and 18.4%, respectively, in acetone-butanol-ethanol fermentations from sugar mixtures of fructose/glucose (4:1) and xylose/glucose (1:2) simulating the hydrolysates of Jerusalem artichoke tubers and corn stover. The results obtained from transcriptional analysis revealed that zinc may have regulatory mechanisms for the sugar transport and metabolism of Clostridium acetobutylicum L7. Therefore, micronutrient zinc supplementation could be an effective way for economic development of butanol production derived from these low-cost agricultural feedstocks.

Assessment of efficacy of specialized food products with modified carbohydrate profile in patients with type 2 diabetes.

Objective - To evaluate the effect of diet therapy with the inclusion of specialized food product with a modified carbohydrate profile (fruit jelly) on glycemic and metabolic parameters in patients with type 2 diabetes mellitus. The study included 45 patients (aged 35 to 69 years) with type 2 diabetes with concomitant obesity I-III degree. The study was conducted in two stages. The first phase evaluated the effect of jelly intake on postprandial glycemia with determination of blood glucose levels on an empty stomach and 30, 60, 120 and 180 minutes after consumption of fruit jelly with maltitol, pectin and sublimated raspberry juice and traditional marmalade with fructose (control), containing 25 g of digestible carbohydrates. In the second phase the effectiveness of combination therapy during 2 weeks of the inclusion of fruit jelly with a modified carbohydrate profile in a standard hypocaloric diet (1550 kcal/day) has been assessed. It has been shown that the dynamics of postprandial glycemia in patients with type 2 diabetes after the consumption of fruit jelly with a modified carbohydrate profile was comparable to the dynamics of blood glucose after eating traditional marmalade with fructose. Inclusion of fruit jelly with a modified carbohydrate profile in the standard diet therapy of patients with type 2 diabetes during 2 weeks was accompanied by positive dynamics of carbohydrate and lipid metabolism, as well as a decrease in the content of lipid peroxidation products in blood plasma: reducing the level of conjugated dienes and malondialdehyde averaged 15.2 and 17.4% from baseline (p<0.05), respectively, while content of lipid peroxidation products in blood plasma did not changed in patients from the control group.

Techno-economic evaluation of conditioning with sodium sulfite for bioethanol production from softwood.

Conditioning with reducing agents allows alleviation of inhibition of biocatalytic processes by toxic by-products generated during biomass pretreatment, without necessitating the introduction of a separate process step. In this work, conditioning of steam-pretreated spruce with sodium sulfite made it possible to lower the yeast and enzyme dosages in simultaneous saccharification and fermentation (SSF) to 1g/L and 5FPU/g WIS, respectively. Techno-economic evaluation indicates that the cost of sodium sulfite can be offset by benefits resulting from a reduction of either the yeast load by 0.68g/L or the enzyme load by 1FPU/g WIS. As those thresholds were surpassed, inclusion of conditioning can be justified. Another potential benefit results from shortening the SSF time, which would allow reducing the bioreactor volume and result in capital savings. Sodium sulfite conditioning emerges as an opportunity to lower the financial uncertainty and compensate the overall investment risk for commercializing a softwood-to-ethanol process.

Antipsychotic Cardiometabolic Side Effect Monitoring in a State Community Mental Health System.

Antipsychotic medications can cause serious cardiometabolic side effects. No recent research has broadly evaluated monitoring and strategies to improve monitoring in U.S. public mental health systems. To address this knowledge gap, we evaluated education with audit and feedback to leaders to improve cardiometabolic monitoring in a state mental health system. We used Chi square statistics and logistic regressions to explore changes in monitoring recorded in randomly sampled records over 2 years. In 2009, assessment of patients on antipsychotics was 29.6 % for cholesterol, 40.4 % for glucose, 29.1 % for triglycerides, 54.3 % for weight, 33.6 % for blood pressure, and 5.7 % for abdominal girth. In 2010, four of ten mental health centers improved their rate of adult laboratory monitoring. Overall monitoring in the state did not increase. Education for prescribers with audit and feedback to leaders can improve monitoring in some settings, but more intensive and/or prolonged interventions may be required.

Conversion of agroindustrial residues for high poly(γ-glutamic acid) production by Bacillus subtilis NX-2 via solid-state fermentation.

Poly(γ-glutamic acid) (γ-PGA) production by Bacillus subtilis NX-2 was carried out through solid-state fermentation with dry mushroom residues (DMR) and monosodium glutamate production residues (MGPR; a substitute of glutamate) for the first time. Dry shiitake mushroom residue (DSMR) was found to be the most suitable solid substrate among these DMRs; the optimal DSMR-to-MGPR ratio was optimized as 12:8. To increase γ-PGA production, industrial waste glycerol was added as a carbon source supplement to the solid-state medium. As a result, γ-PGA production increased by 34.8%. The batch fermentation obtained an outcome of 115.6 g kg(-1) γ-PGA and 39.5×10(8) colony forming units g(-1) cells. Furthermore, a satisfactory yield of 107.7 g kg(-1) γ-PGA was achieved by compost experiment on a scale of 50 kg in open air, indicating that economically large-scale γ-PGA production was feasible. Therefore, this study provided a novel method to produce γ-PGA from abundant and low-cost agroindustrial residues.

Assessment of integrated process based on hydrothermal and alkaline treatments for enzymatic saccharification of sweet sorghum stems.

In this study, sweet sorghum stem was subjected to hydrothermal pretreatment (HTP) and alkaline post-treatment to enhance its saccharification ratio by reducing its recalcitrance. The results showed that the HTP (110-210°C, 0.5-2.0h) significantly degraded hemicelluloses, and the pretreatment at the temperature higher than 190°C led to the partial degradation of the cellulose. As compared to the sole HTP, the integrated process removed most of lignin and hemicelluloses, which incurred a higher cellulose saccharification ratio. Under an optimum condition evaluated (HTP at 170°C for 0.5h and subsequent 2% NaOH treatment), 77.5% saccharification ratio was achieved, which was 1.8, 2.0 and 5.5 times as compared to the only HTP pretreated substrates, alkaline treated substrates alone and the raw material without pretreatment, respectively. Clearly, the integrated process can be considered as a promising approach to achieve an efficient conversion of lignocellulose to fermentable glucose.

Efficient production of l-lactic acid using co-feeding strategy based on cane molasses/glucose carbon sources.

L-Lactic acid is an important platform chemical, which ought to be produced under cost control to meet its huge demand. Cane molasses, a waste from sugar manufacturing processes, is hopeful to be utilized as a cheap carbon source for L-lactic acid fermentation. Considering that cane molasses contains nutrients and hazardous substances, efficient production of L-lactic acid was developed by using a co-feeding strategy based on the utilization of cane molasses/glucose carbon sources. Based on the medium optimization with response surface method, 168.3g/L L-lactic acid was obtained by a Bacillus coagulans strain H-1 after 78h fed-batch fermentation, with a productivity of 2.1g/Lh and a yield of 0.88g/g. Since cane molasses is a feasible carbon source, the co-feeding fermentation might be a promising alternative for the economical production of L-lactic acid.

Assessment of insulin action on carbohydrate metabolism: physiological and non-physiological methods.

Carbohydrate metabolism in humans is regulated by insulin secretion from pancreatic ß-cells and glucose disposal by insulin-sensitive tissues. Insulin facilitates glucose utilization in peripheral tissues and suppresses hepatic glucose production. Any defects in insulin action predispose an individual to glucose intolerance and Type 2 diabetes mellitus. Early detection of defects in insulin action could provide opportunities to prevent or delay progression of the disease state. There are different approaches to assess insulin action. Initial methods, such as peripheral insulin concentration and simple indices, have several limitations. Subsequently, researchers developed methodologies using intravenous glucose infusion to determine glucose fluxes. However, these methodologies are limited by being non-physiological. Newer, innovative techniques that have been developed are more sophisticated and physiological. By modelling glucose kinetics using isotope dilution techniques, several robust parameters can be obtained that are physiologically relevant and sound. This brief review summarizes most of the non-physiological and physiological methodologies used to measure the variables of insulin action.

Cost analysis of cassava cellulose utilization scenarios for ethanol production on flowsheet simulation platform.

Cassava cellulose accounts for one quarter of cassava residues and its utilization is important for improving the efficiency and profit in commercial scale cassava ethanol industry. In this study, three scenarios of cassava cellulose utilization for ethanol production were experimentally tested under same conditions and equipment. Based on the experimental results, a rigorous flowsheet simulation model was established on Aspen plus platform and the cost of cellulase enzyme and steam energy in the three cases was calculated. The results show that the simultaneous co-saccharification of cassava starch/cellulose and ethanol fermentation process (Co-SSF) provided a cost effective option of cassava cellulose utilization for ethanol production, while the utilization of cassava cellulose from cassava ethanol fermentation residues was not economically sound. Comparing to the current fuel ethanol selling price, the Co-SSF process may provide an important choice for enhancing cassava ethanol production efficiency and profit in commercial scale.

Consumption of fructose-sweetened beverages for 10 weeks reduces net fat oxidation and energy expenditure in overweight/obese men and women.

BACKGROUND/OBJECTIVES: The results of short-term studies in humans suggest that, compared with glucose, acute consumption of fructose leads to increased postprandial energy expenditure and carbohydrate oxidation and decreased postprandial fat oxidation. The objective of this study was to determine the potential effects of increased fructose consumption compared with isocaloric glucose consumption on substrate utilization and energy expenditure following sustained consumption and under energy-balanced conditions. SUBJECTS/METHODS: As part of a parallel arm study, overweight/obese male and female subjects, 40-72 years, consumed glucose- or fructose-sweetened beverages providing 25% of energy requirements for 10 weeks. Energy expenditure and substrate utilization were assessed using indirect calorimetry at baseline and during the 10th week of intervention. RESULTS: Consumption of fructose, but not glucose, led to significant decreases of net postprandial fat oxidation and significant increases of net postprandial carbohydrate oxidation (P<0.0001 for both). Resting energy expenditure (REE) decreased significantly from baseline values in subjects consuming fructose (P=0.031) but not in those consuming glucose. CONCLUSIONS: Increased consumption of fructose for 10 weeks leads to marked changes of postprandial substrate utilization including a significant reduction of net fat oxidation. In addition, we report that REE is reduced compared with baseline values in subjects consuming fructose-sweetened beverages for 10 weeks.

Toxic impact of two organophosphate insecticides on biochemical parameters of a food fish and assessment of recovery response.

Sublethal effects of chlorpyrifos (CPF) and monocrotophos (MCP) on fish biochemical constituents were investigated along with the assessment of recovery response after cessation of intoxication. The fish, Clarias batrachus were exposed to 1.656 mg(-l) and 2.114 mg(-l) of CPF and MCP for 28 days. After 28 days, they were released in freshwater and allowed to recover for 21 days. The CPF exposure resulted in the decrease of carbohydrate and glycogen content, whereas MCP intoxication caused mixed response. Pyruvate and lactate contents were altered under the stress of CPF and MCP. Recovery of these alterations was observed after the cessation of toxicity. Exposure of C. batrachus to CPF and MCP resulted in decreased activity of lactate dehydrogenase in the kidney, liver and muscle but its activity increased in the gills. The CPF caused inhibition of succinate dehydrogenase enzyme in all tissues. Induction in the activity of malate dehydrogenase was caused by both insecticides. Glycogen phosphorylase a was induced in all tissues, whereas glycogen phosphorylase ab showed both induction and inhibition. Of the two insecticides, CPF was more toxic and the recovery response was less. These results are important in the assessment of the risk caused by organophosphate insecticides on nontarget organisms, especially the food fish.

Increasing protein at the expense of carbohydrate in the diet down-regulates glucose utilization as glucose sparing effect in rats.

High protein (HP) diet could serve as a good strategy against obesity, provoking the changes in energy metabolic pathways. However, those modifications differ during a dietary adaptation. To better understand the mechanisms involved in effect of high protein diet (HP) on limiting adiposity in rats we studied in parallel the gene expression of enzymes involved in protein and energy metabolism and the profiles of nutrients oxidation. Eighty male Wistar rats were fed a normal protein diet (NP, 14% of protein) for one week, then either maintained on NP diet or assigned to a HP diet (50% of protein) for 1, 3, 6 and 14 days. mRNA levels of genes involved in carbohydrate and lipid metabolism were measured in liver, adipose tissues, kidney and muscles by real time PCR. Energy expenditure (EE) and substrate oxidation were measured by indirect calorimetry. Liver glycogen and plasma glucose and hormones were assayed. In liver, HP feeding 1) decreased mRNA encoding glycolysis enzymes (GK, L-PK) and lipogenesis enzymes(ACC, FAS), 2) increased mRNA encoding gluconeogenesis enzymes (PEPCK), 3) first lowered, then restored mRNA encoding glycogen synthesis enzyme (GS), 4) did not change mRNA encoding ß-oxidation enzymes (CPT1, ACOX1, ßHAD). Few changes were seen in other organs. In parallel, indirect calorimetry confirmed that following HP feeding, glucose oxidation was reduced and fat oxidation was stable, except during the 1(st) day of adaptation where lipid oxidation was increased. Finally, this study showed that plasma insulin was lowered and hepatic glucose uptake was decreased. Taken together, these results demonstrate that following HP feeding, CHO utilization was increased above the increase in carbohydrate intake while lipogenesis was decreased thus giving a potential explanation for the fat lowering effect of HP diets.

Arabidopsis plants acclimate to water deficit at low cost through changes of carbon usage: an integrated perspective using growth, metabolite, enzyme, and gene expression analysis.

Growth and carbon (C) fluxes are severely altered in plants exposed to soil water deficit. Correspondingly, it has been suggested that plants under water deficit suffer from C shortage. In this study, we test this hypothesis in Arabidopsis (Arabidopsis thaliana) by providing an overview of the responses of growth, C balance, metabolites, enzymes of the central metabolism, and a set of sugar-responsive genes to a sustained soil water deficit. The results show that under drought, rosette relative expansion rate is decreased more than photosynthesis, leading to a more positive C balance, while root growth is promoted. Several soluble metabolites accumulate in response to soil water deficit, with K(+) and organic acids as the main contributors to osmotic adjustment. Osmotic adjustment costs only a small percentage of the daily photosynthetic C fixation. All C metabolites measured (not only starch and sugars but also organic acids and amino acids) show a diurnal turnover that often increased under water deficit, suggesting that these metabolites are readily available for being metabolized in situ or exported to roots. On the basis of 30 enzyme activities, no in-depth reprogramming of C metabolism was observed. Water deficit induces a shift of the expression level of a set of sugar-responsive genes that is indicative of increased, rather than decreased, C availability. These results converge to show that the differential impact of soil water deficit on photosynthesis and rosette expansion results in an increased availability of C for the roots, an increased turnover of C metabolites, and a low-cost C-based osmotic adjustment, and these responses are performed without major reformatting of the primary metabolism machinery.

Effluent impact assessment using microarray-based analysis in common carp: a systems toxicology approach.

Effluents are a main source of direct and continuous input of pollutants to the aquatic environment, and can cause ecotoxicological effects at different levels of biological organization. Since gene expression responses represent the primary interaction site between environmental contaminants and biota, they provide essential clues to understand how chemical exposure can affect organismal health. The aim of the present study was to investigate the applicability of a microarray approach for unraveling modes of action of whole effluent toxicity and impact assessment. A chronic toxicity test with common carp (Cyprinus carpio) was conducted where fish were exposed to a control and 100% effluent for 21 days under flow-through conditions. Microarray analysis revealed that effluent treatment mainly affected molecular pathways associated with the energy balance of the fish, including changes in carbohydrate and lipid metabolism, as well as digestive enzyme activity. These gene expression responses were in clear agreement with, and provided additional mechanistic information on various cellular and higher level effects observed for the same effluent. Our results demonstrate the benefit of toxicogenomic tools in a "systems toxicology" approach, involving the integration of adverse effects of chemicals and stressors across multiple levels of biological complexity.

Anticipated yield loss in field-grown soybean under elevated ozone can be avoided at the expense of leaf growth during early reproductive growth stages in favourable environmental conditions.

Ozone is a powerful oxidizing agent which is responsible for more damage to vegetation than any other air pollutant. In this study, leaf growth, photosynthesis, and carbohydrate content were analysed during the seed-filling growth stage of field-grown soybeans exposed to ambient air and 1.2 times ambient ozone concentration using a Free Air Concentration Enrichment (FACE) facility. By contrast to predictions based on controlled-environment and open-top chamber studies, final yield did not differ between treatments, although the cultivar used here was sensitive to ozone damage: growth and carbohydrate content of upper canopy leaves was reduced during the seed-filling stage in which an ozone-induced decrease of photosynthesis was present. However, 2004 was an ideal growing season in central Illinois and the cumulative ozone indices were lower than in previous years. Still, the results indicate that the anticipated yield loss under ozone concentrations was avoided at the expense of leaf growth, as reserves were diverted from vegetative to reproductive organs.