Enzyme leakage during cryopreservation of ram spermatozoa

Spermatozoal motility and enzyme leakage during cryopreservation of ram spermatozoa was evaluated in the presence and absence of antifreeze proteins (AFP). Loss of spermatozoal motility due to the cryopreservation process was reduced by 10% in the presence of AFP. Samples of the diluted semen at various stages of the cryopreservation process were assayed for aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), acid phosphatase (ACP) and lactate dehydrogenase (LDH). The levels of AST, ALT and ACP were low in ram spermatozoa and were not considered suitable for the objective measurement of spermatozoal damage. ALP leaked during the cooling and freezing process, whereas LDH leakage was prevalent during cooling, freezing and post-thaw incubation of spermatozoa. Changes in ALP and LDH could be used as marker enzymes in the development of semen processing protocols and of semen diluents.     

ZnO nanoparticles augment ALT,AST, ALP and LDH expressions in C2C12 cells

The present study aimed to investigate the effect of ZnO nanoparticles on alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) enzyme expressions in C2C12 cells. ZnO nanoparticles are widely used in the several cosmetic lotions and other biomedical products. Several studies report on ZnO nanoparticle mediated cytotoxicity. However, there are no reports on the effect of ZnO nanoparticles on ALT, AST, ALP and LDH enzyme expressions in C2C12 cells. A cytotoxicity assay was carried out to determine the effect of ZnO nanoparticles (1–5 mg/ml) on C2C12 cell viability at 48 and 72 h. ZnO nanoparticles increased ALT, AST, ALP and LDH enzyme mRNA expression and their activities in C2C12 cells. In conclusion, the present study showed that ZnO nanoparticles increased these enzyme activities and its mRNA expression in C2C12 cells in a dose-dependent manner.     

Effect of a polyherbal formulation, Ambrex, on butylated hydroxy toluene (BHT) induced toxicity in rats

Effect of polyherbal formulation Ambrex was evaluated in butylated hydroxytoluene (BHT) induced toxicity of lungs and liver in rats. Toxicity was produced by administering BHT (500 mg/kg/day) for 3 days. Lung damage was evidenced by elevated levels of broncho alveolar lavage fluid (BAL) parameters such as protein, lactate, lactate dehydrogenase (LDH), alkaline phosphatase (ALP), acid phosphatase (ACP) and glucose-6-phosphate dehydrogenase (G6PDH). Liver damage was proved by elevated levels of serum protein and markers such as LDH, ALP, aspartate amino transferase (AST), alanine amino transferase (ALT), decreased level of lipid peroxides (LPO) in serum and glutathione (GSH) in liver. Administration of aqueous suspension of Ambrex (50 mg/kg orally) retained these elevated levels of BAL-protein, lactate, LDH, ALP, ACP, G6PDH and serum-protein, LDH, ALP, AST and ALT at near normal values. Decreased level of liver GSH was retained at near normalcy in Ambrex pretreated BHT-administered animals. There was no change in liver LPO in all the four groups.     

L-Gln and L-Ser suppress the D-galactosamine-induced IL-18 expression and hepatitis

d-Galactosamine (GalN) induces acute hepatitis in experimental animals and this hepatitis has been shown to be suppressed by preceding ingestion of amino acids such as Gly, l-Ser, and l-Gln. However, little is known about the mechanism of its action. The present study shows for the first time that IL-18 reduction is involved in the suppressive actions of l-Gln and l-Ser on GalN-induced hepatitis. Elevation of IL-18 mRNA expression in liver and its concentration in serum in GalN-treated rats were found to be suppressed by preceding ingestion of 10% l-Gln- or 10% l-Ser diets, and resulted in the attenuation of the increase in serum transaminase (ALT and AST) activities, indexes of hepatic injury. These results suggest that suppressive effects of some dietary amino acids on the GalN-induced hepatitis are mediated by IL-18 reduction.     

Zinc supplementation and cardiovascular disease risk factors: A GRADE-assessed systematic review and dose-response meta-analysis

Background and objectiveA deficit in zinc has been related to a higher probability of developing cardiovascular diseases (CVDs). The anti-inflammatory and anti-oxidative capabilities of zinc may have a wide range of therapeutic impacts on CVDs. We conducted a comprehensive systematic review and meta-analysis of the possible impacts that zinc supplementation may have on the risk factors associated with CVDs.MethodsTo identify eligible randomized clinical trials (RCTs) evaluating the effects of zinc supplementation on CVDs risk factors, electronic databases including PubMed, Web of Science, and Scopus were systematically searched up to January 2023. The heterogeneity of trials was checked using the I² statistic. According to the heterogeneity tests, random effects models were estimated and pooled data were defined as the weighted mean difference (WMD) with a 95% confidence interval (CI).ResultsOf 23165 initial records, 75 studies that met inclusion criteria were analyzed in this meta-analysis. The pooled findings indicated the significant lowering effects of zinc supplementation on triglycerides (TG), total cholesterol (TC), fasting blood glucose (FBG), Hemoglobin A1C (HbA1C), Homeostatic Model Assessment for Insulin Resistance (HOMA-IR), C-reactive protein (CRP), interleukin-6 (IL-6), Tumor necrosis factor-α (TNF-α), nitric oxide (NO), malondialdehyde (MDA), total antioxidant capacity (TAC), and glutathione (GSH), with no noticeable effects on low-density lipoprotein (LDL), high-density lipoprotein (HDL), insulin, systolic blood pressure (SBP), diastolic blood pressure (DBP), aspartate transaminase (AST), and Alanine aminotransferase (ALT).ConclusionOverall, zinc supplementation may boost recognized coronary risk factors that contribute to the development of CVDs. Future research should be conducted to bolster our results.     

Seasonal and physiological variations in serum chemistry and mineral concentrations in cattle

The present study reports the seasonal and physiological variations of copper, zinc, magnesium, iron sodium chlorine, potassium, calcium, phosphorus, urea, alkaline phosphatase (ALP), creatinine (CR), aspartate aminotransfrase (AST), alanine aminotransferase (ALT), cholesterol, albumin, globulin, lactate dehydrogenase (LDH), and total protein concentrations in cattle. Two groups of mated (n=14) and nonmated (n=10), healthy cows were selected for the study. Serum samples were collected at each of four periods: (1) early pregnancy (May), (2) midpregnancy (August), (3) late pregnancy (October), and (4) lactation (February). Physiological variations result in changes of cholesterol, calcium, LDH, and total protein concentrations. Phosphorus varies only with seasonal but not physiological changes, whereas ALP, copper, magnesium, and potassium concentrations change with physiological and seasonal conditions. The copper concentration is increased through the pregnancy. Neither the seasonal nor the physiologic variations affect zinc, iron, sodium, chlorine, calcium, urea, creatinine, albumin, and globulin values in both groups in all periods. Thus, these values can be used as reference for both mater and nonmated bovines. The measured total protein might not reflect its true value because of dehydration during the hot season. These observations suggest that seasonal and physiologic variations have to be taken into consideration for the correct interpretation of serum chemistry and elements status in cattle. Nutritional supplements are required for cattle during certain periods to avoid a decline of their performance, which would then represent consequent economic loses.     

支链氨基酸对运动大鼠氨基酸代谢和运动能力的影响

观察了支链氨基酸(BCAA)对大鼠运动能力和血清游离氨基酸代谢的影响。实验用21只雄性wistar大鼠,随机分为3组:正常组、游泳对照组和游泳补充BCAA组。2个运动组每天游泳训练1h,10天后游泳6h,观察补充BCAA对大鼠游泳运动能力和血清游离氨基酸水平的影响。实验结果表明,补充BCAA可明显提高大鼠游泳存活率,抑制血清中必需氨基酸、非必需氨基酸和总氨基酸水平升高,游泳运动后大鼠的血清中乳酸和LDH的升高幅度有所降低,抑制骨骼肌LDH活力的下降。说明补充BCAA可明显提高大鼠的运动能力,减少运动造成的蛋白质分解     

Influence of dietary culture material containing aflatoxin and T<Subscript>2</Subscript> toxin on certain serum biochemical constituents in Japanese quail

Forty newly hatched unsexed Japanese quail (Coturnix coturnix japonica) chicks were fed diets containing 3 ppm aflatoxin (AF) and 4 ppm T₂ toxin either singly and in combination for 35 days. Sera samples were collected from six birds in each group at the end of the trial to study the effect of certain serum biochemical parameters. Highly significant (P < 0.01) differences were observed for serum total protein, albumin, globulin, glucose, cholesterol, aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP) and gamma glutamyl transferase (GGT) between control and toxin treated groups. Toxin treated groups did not reveal any significant difference for serum urea nitrogen, creatinine, uric acid, calcium, phosphorus, sodium and potassium. There was a reduction in the levels of serum total protein, albumin, globulin, glucose, cholesterol, ALT and elevation of AST and GGT and variable ALP levels observed in toxin treated groups. Forms part of M.V.Sc. thesis of first author approved by the Tamil Nadu Veterinary and Animal Sciences University, Chennai 600 051, India.     

Biochemical predictors for Sars-Cov-2 severity

It is of interest to assess the inflammatory marker profile in SARS-CoV-2 patients and to correlate the levels of systemic inflammatory biomarkers such as neutrophil-to-lymphocyte ratio (NLR), C-Reactive Protein CRP, Ferritin, Creatine kinase (CK), Lactate dehydrogenase (LDH) and liver function analytes total serum proteins, albumin, total bilirubin, direct bilirubin, alkaline phosphatase, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) with the severity of SARS-CoV-2 infections. A total of 1000 COVID-19 positive patient''s data were collected. Laboratory assessments consisted of NLR (neutrophil-lymphocyte ratio) by cell counter, C Reactive Protein (CRP) by immunoturbidimetry, Ferritin by electrochemiluminescence (ECLIA) and Creatine Kinase (CK), Lactate Dehydrogenase (LDH), Alkaline phosphatase (ALP), Aspartate aminotransferase (AST), Alanine aminotransferase (ALT), Total Bilirubin, Direct Bilirubin, Total Protein and Albumin by spectrophotometry. The mean plasma CRP levels, NLR, ferritin, CK and LDH levels were higher in severe cases than in non-severe cases, and the difference was statistically significant (p<0.05). All liver function tests such as the total and direct bilirubin, AST, ALT, ALP, total protein and albumin were higher in severe patients than non-severe patients and the difference was statistically significant (p<0.05). Data indicate that NLR, CRP, Ferritin, CK, LDH and liver function analytes have a crucial role as prognostic markers for SARS-CoV-2 infections and hence should be routinely recommended for risk assessment and stratification of the patients to reduce the associated morbidity and mortality.     

The effect of citric acid and microbial phytase on mineral utilization in broiler chicks

An experiment was conducted to study the effect of microbial phytase (Natuphos^® 500) supplementation in chicks fed different levels of available phosphorus (AP) and citric acid (CA) on performance, mineral retention (Ca, P, Mg, and Zn), bone and plasma minerals (Ca, P, Mg, and Zn), plasma total protein (TP), and serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH) activities. Data were analyzed as a 2×4×2 factorial arrangement with two levels of AP (3.5 and 2.5 g/kg), four levels of phytase (0, 200, 400 and 600 U/kg), and two levels of citric acid (0 and 20 g/kg). The low-AP diets reduced performance. Phytase supplementation increased weight gain (up to 7% quadratically) and feed consumption (up to 5%). This response was statistically maximized by 200 U/kg phytase. Feed to gain ratio was not affected by phytase addition. Growth response to phytase was negatively affected by citric acid. Decreasing AP content in the diet increased Ca, P, and Mg retention, and reduced Zn retention. Phytase supplementation linearly increased Ca, P, and Zn retention by 9, 10 and 16%, respectively. Citric acid addition also increased Ca, P, and Zn retention by 3, 3 and 4%, respectively. Likewise, the decrease in AP content in the diet caused a reduction of tibia ash and tibia Zn, and an increase in tibia Ca and P contents. Phytase supplementation increased tibia ash (up to 4%), tibia Ca (up to 2%), P (up to 1%) and Zn (up to 4%) contents, tibia weight (up to 9%), and relative tibia (up to 19%) and liver (up to 13%) weights. Citric acid increased tibia ash (2%), and tibia Ca (2%) and P (2%) contents. Finally, by decreasing AP levels in the diet, plasma Ca and Zn concentrations as well as AST, ALP, and LDH activities were increased. However, plasma P and TP content were reduced. Phytase supplementation increased linearly plasma Ca (up to 4%), P (up to 12%), Mg (up to 10%), Zn (up to 22%) and TP (up to 7%) content, and serum AST (up to 22%), ALT (up to 40%), and LDH (up to 17%) activities, and reduced linearly serum ALP (up to 34%) activity. Citric acid addition increased plasma Ca, Mg, and Zn content by 10, 4, and 5%, respectively, and reduced ALP activity by 13%. In conclusion, these results indicated that the addition of phytase to maize and soyabean meal low-AP diets improved the performance and increased Ca, P, and Zn utilization in chicks. However, the inclusion of citric acid depressed the performance and caused an increase in mineral utilization. Growth response to phytase was negatively affected by citric acid.     

Potential preventive effect of carvacrol against diethylnitrosamine-induced hepatocellular carcinoma in rats

Antioxidants are one of the key players in tumorigenesis, several natural and synthetic antioxidants were shown to have anticancer effects. The aim of the present study is to divulge the chemopreventive nature of carvacrol during diethylnitrosamine (DEN)-induced liver cancer in male wistar albino rats. Administration of DEN to rats resulted in increased relative liver weight and serum marker enzymes aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), and gamma glutamyl transpeptidase (γGT). The levels of lipid peroxides elevated (in both serum and tissue) with subsequent decrease in the final body weight and tissue antioxidants like superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH), glutathione peroxidase (GPx), and glutathione reductase (GR). Carvacrol supplementation (15 mg/kg body weight) significantly attenuated these alterations, thereby showing potent anticancer effect in liver cancer. Histological observations and transmission electron microscopy studies were also carried out, which added supports to the chemopreventive action of the carvacrol against DEN-induction during liver cancer progression. These findings suggest that carvacrol prevents lipid peroxidation, hepatic cell damage, and protects the antioxidant system in DEN-induced hepatocellular carcinogenesis.     

A novel amino acid supplementation strategy based on a stoichiometric model to enhance human IL-2 (interleukin-2) expression in high-cell-density Escherichia coli cultures

A novel amino acid supplementation strategy was developed for enhancing the production of IL-2 (interleukin-2; as a model protein) by recombinant Escherichia coli BL21 (pET21a-hil2) in fed-batch high-cell-density cultures. The amino acids most needed and their amounts were determined using a stoichiometric model, and full factorial design experiments were conducted to determine the effects of single amino acids and amino acid mixtures on production. One of the most effective amino acid mixtures was found to be leucine, aspartic acid and glycine. This amino acid mixture was utilized for the production of IL-2 in batch and fed-batch fermentations. The amount of IL-2 produced increased from 403 to 722 mg/l and from 5.15 × 103 to 8.08 × 103 mg/l in batch and fed-batch cultures respectively. The results also revealed that the above amino acid mixture specifically increases IL-2 concentration in the cells.     

Effects of Exercise and Zinc Supplementation on Cytokine Release in Young Wrestlers

The present study aims to examine the effect of zinc supplementation on the release of some cytokines in young wrestlers actively involved in wrestling. A total of 40 male subjects of the same age group were included in the study: half were wrestlers and the other half were not involved in sports. The subjects were equally divided into four groups and treated during an 8-week period as follows: group 1, zinc-supplemented athletes; group 2, non-supplemented athletes; group 3, zinc-supplemented sedentary subjects, and group 4, non-supplemented sedentary group. Blood samples were taken from each subject at the beginning and at the end of the study period. The serum tumor necrosis factor-α (TNF-α), interleukin-2 (IL-2), and interpheron-γ levels (IFN-γ) were determined using the enzyme-linked immunosorbent assay method. At the beginning of the study, there were no significant differences of the measured parameters between the four study groups. At the end of the study, the levels of TNF-α, IL-2, and IFN-γ were significantly higher in the two zinc-supplemented groups compared to those that did not receive supplementation, regardless of the activity status (p < 0.01).     

Gastroprotective activity and mechanism of novel dichlorido-zinc(II)-4-(2-(5-methoxybenzylideneamino)ethyl)piperazin-1-iumphenolate complex on ethanol-induced gastric ulceration

Zinc complexes were reported to have anti-ulcer activity and used as drug for the treatment of gastrointestinal disorders. A novel compound dichlorido-zinc(II)-4-(2-(5-methoxybenzylidene amino)ethyl)piperazin-1-iumphenolate (ZnHMS) was synthesized, characterized and evaluated for its gastroprotective activity against ethanol-induced ulcer in rats. Gross and microscopic lesions, histochemical staining of glycogen storage, biochemical and immunological parameters were taken into consideration. Oral administration of ZnHMS (30 and 60 mg/kg; 14 days) dose-dependently inhibited gastric lesions. It significantly increased the mucus content and total acidity compared to the control group (P<0.01). Serum levels of aspartate (AST), alanine (ALT) transaminases, pro-inflammatory interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α) and anti-inflammatory interleukin-10 (IL-10) in the rats exposed to ethanol induced ulceration have been altered. ZnHMS considerably enhances (P<0.05) the protection of gastric epithelia by modulating the acute alterations of AST, ALT, IL-6, IL-10, TNF-α and stomach glycogen. Interestingly, ZnHMS did interfere with the natural release of nitric oxide. In addition, acute toxicity study revealed no abnormal sign to the rats treated with ZnHMS (2000 mg/kg). These findings suggest that the gastroprotective activity of ZnHMS might contribute in adjusting the inflammatory cytokine-mediated oxidative damage to the gastric mucosa.     

Coordinated increase in albumin, fibrinogen, and muscle protein synthesis during hemodialysis: role of cytokines

Serum albumin, fibrinogen levels, and lean body mass are important predictors of outcome in end-stage renal disease (ESRD). We estimated the fractional synthesis rates of albumin (FSR-A), fibrinogen (FSR-F), and muscle protein (FSR-M) in nine ESRD patients and eight controls, using primed constant infusion of l-[ring-(13)C(6)]phenylalanine. Cytokine profile and arteriovenous balance of amino acids were also measured. ESRD patients were studied before (Pre-HD) and during hemodialysis (HD). Plasma IL-6, IL-10, and C-reactive protein increased significantly during HD. Despite a decrease in the delivery of amino acids to the leg, the outflow of the amino acids increased during HD. The net balance of amino acids became more negative during HD, indicating release from the muscle. HD increased leg muscle protein synthesis (45%) and catabolism (108%) but decreased whole body proteolysis (15%). FSR-A during HD (9.7 +/- 0.9%/day) was higher than pre-HD (6.5 +/- 0.9%/day) and controls (5.8 +/- 0.5%/day, P < 0.01). FSR-F increased during HD (19.7 +/- 2.6%/day vs. 11.8 +/- 0.6%/day, P < 0.01), but it was not significantly different from that of controls (14.4 +/- 1.4%/day). FSR-M intradialysis (1.77 +/- 0.19%/day) was higher than pre-HD (1.21 +/- 0.25%/day) and controls (1.30 +/- 0.32%/day, P < 0.001). Pre-HD FSR-A, FSR-F, and FSR-M values were comparable to those of controls. There was a significant and positive correlation between plasma IL-6 and the FSRs. Thus, in ESRD patients without metabolic acidosis, the fractional synthesis rates of albumin, fibrinogen, and muscle protein are not decreased pre-HD. However, HD increases the synthesis of albumin, fibrinogen, and muscle protein. The coordinated increase in the FSRs is facilitated by constant delivery of amino acids derived from the muscle catabolism and intradialytic increase in IL-6.     

Protective effects of dl-α-tocopherol acetate and sodium selenate on the liver of rats exposed to gamma radiation

The aim of this study is to investigate whether vitamin E (as dl-α-tocopherol acetate) and selenium (as sodium selenate) exert a protective effect against radiation damage. The liver tissue of rats irradiated with a single dose of 1000 cGy ⁶⁰Co-γ-irradiation was examined for morphological changes after the intraperitoneal (ip) administration dl-α-tocopherol acetate and sodium selenate as compared to controls. Also, the amounts of blood glutathione and serum alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), and total protein were determined by spectrophotometric methods. Degenerative changes were observed under light and electron microscopy in the liver tissue of the control (radiation only) group. In the group receiving radiation and ip doses of dl-α-tocopherol acetate and sodium selenate, the damage to the liver tissue was minimal or absent. In the radiation-only group, a reduction of the blood glutathione level and increases in serum values of AST, ALT, ALP, and LDH activity were observed, whereas in the irradiation-treated group, the reverse was found to occur. Based on these morphological and biochemical observations, it was concluded that the ip administration of dl-α-tocopherol acetate and sodium selenate exerts a protective effect against liver radiation damage.     

Efficacy of Citrus reticulata and Mirazid in treatment of Schistosoma mansoni

This work has been carried out to investigate the effect of Schistosoma mansoni infection on mice livers after treatment with the ethanolic extract of Citrus reticulata root or the oleo-resin extract from Myrrh of Commiphora molmol tree (Mirazid), as a new antishistosomal drug. Marker enzymes for different cell organelles were measured; succinate dehydrogenase (SDH); lactate dehydrogenase (LDH) and its isoenzymes; glucose-6-phosphatase (G-6-Pase); acid phosphatase (AP) and 5'- nucleotidase. Liver function enzymes; aspartate aminotransferase (AST); alanine aminotransferase (ALT), and alkaline phosphatase (ALP) were also estimated. Parasitological studies through ova count and worm burden will also be taken into consideration. The results showed a marked reduction in SDH, LDH, AST, and ALT enzyme activities and a significant increase in G-6-Pase, AP, 5'- nucleotidase, and ALP after S. mansoni infection. A noticeable alteration in LDH subunits were also noticed. Treatment with C. reticulata or Mirazid improved all the previous enzyme activities with a noticeable reduction in ova count and worm burden.     

Smad3基因剔除小鼠血清酶活性的变化

目的　探讨Smad3基因对剔除小鼠血清酶活性的影响。方法　采用荷兰半自动生化分析仪对 35日龄、70日龄、6月龄的三种不同基因型的小鼠的ALP、AST、ALT、CK、LDH L进行测定。结果　纯合型小鼠各项指标较野生型高 ,且表现出ALP随着年龄的增长而降低 ;AST、ALT、CK、LDH L随着年龄的增长而增高。结论　Smad3基因的剔除对小鼠的血清酶活性有一定的影响 ,为该小鼠的进一步研究提供基础。     

Enzyme activities in plasma, liver and kidney of black ducks and mallards

Activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatine phosphokinase (CPK), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) were measured in plasma, liver, and kidney, and gamma-glutamyl transferase (GGT) was measured in liver and kidney of black ducks (Anas rubripes). Activities of ALT, AST, GGT, and ornithine carbamyl transferase (OCT) were assayed in plasma, liver, and kidney of game-farm mallards (Anas platyrhynchos). Appreciable OCT and AST activity occurred in both liver and kidney. Activities of ALT, CPK, ALP and GGT were higher in kidney, while LDH was higher in liver, GGT was detected in plasma from one of four mallards.     

The effect of fish oil supplementation on cytokine production in children

The ex vivo production of inflammatory cytokines during fish oil supplementation (n-3 polyunsaturated fatty acids, n-3 PUFA) is a matter of considerable controversy. Studies on human subjects have generally reported decreased lymphocyte proliferation and decreased production of IL-2, interferon-gamma, IL-1beta, IL-6 and TNF-alpha, but other studies showed no effect or even increased production. There are no published reports on ex vivo cytokine production in children on long-term, n-3 PUFA supplementation. The current double-blind study explored cytokine production by peripheral blood mononuclear cells (PBMCs), with and without lipopolysaccharide (LPS) stimulation in children on 12 weeks' supplementation with 300 mg/day of n-3 PUFA. Twenty-one children (aged 8-12 years) were randomized to receive 1 g canola oil (control) or 300 mg n-3 PUFA + 700 mg canola oil in a chocolate spread. Blood was then drawn and PBMCs were separated and cultured for 24 h in a culture medium with or without 10 microg/mL LPS for 5 x 10(6) PBMCs. The pro-inflammatory cytokines, IL-1beta, TNF-alpha and IL-6, and the anti-inflammatory cytokines, IL-10 and IL-1RA, were evaluated by ELISA. The levels of all the cytokines were higher in non-stimulated and LPS-stimulated cultures, from n-3 PUFA-treated subjects as compared to controls. There was no difference in the IL-1beta/IL-1RA ratio between the two groups, with and without LPS stimulation. Nevertheless, the ratio tended to be lower in the treated subjects on both occasions. In conclusion, our results indicate an increased production of both pro-inflammatory and anti-inflammatory cytokines, with and without LPS stimulation, in children on 12 weeks' n-3 PUFA supplementation.