Association of interleukin-2 and interleukin-10 with the pathophysiology and development of generalized anxiety disorder: a case-control study.

BACKGROUND: Generalized anxiety disorder (GAD) is a devastating mental health condition characterized by constant, uncontrolled worrying. Recent hypotheses indicate that pro-inflammatory cytokines and chemokines are potential contributors to the pathogenesis of GAD. Here, we aimed to assess the role of interleukin-2 (IL-2) and interleukin-10 (IL-10) in the pathophysiology and development of GAD. METHODS: This study recruited 50 GAD patients diagnosed according to the DSM-5 criteria and 38 age-sex-matched healthy controls (HCs). A qualified psychiatrist evaluated all study subjects. The socio-demographic and clinical characteristics of the study population were determined using pre-structured questionnaires or interviews, and cytokine serum levels were estimated using commercially available ELISA kits. RESULTS: We observed reduced serum IL-10 levels in GAD patients compared to HCs (33.69 ± 1.37 pg/ml vs. 44.12 ± 3.16 pg/ml). Also, we observed a significant negative correlation between altered IL-10 levels and GAD-7 scores (r=-0.315, p = 0.039). Moreover, IL-10 serum measurement exhibited good predictive value in receiver operating characteristics (ROC) analysis with an area under the curve (AUC) value of 0.793 (p < 0.001) with 80.65% sensitivity and 62.79% specificity at a cutoff value of 33.93 pg/ml. Conversely, we noticed elevated serum IL-2 levels in GAD patients than in HCs (14.81 ± 2.88 pg/ml vs. 8.08 ± 1.1 pg/ml); however, it failed to maintain any significant association with GAD-7 scores, implying that IL-2 might not be involved in GAD pathogenesis. The lower AUC value (0.640; p > 0.05) exhibited by IL-2 serum measurement in ROC analysis further supported that IL-2 might not be associated with GAD. CONCLUSION: This study provides new insights into the complex interplay between anti-inflammatory cytokines and GAD pathogenesis. Based on the present findings, we can assume that IL-10 but not IL-2 may be associated with the pathophysiology and development of GAD. However, further research with a larger population size and longitudinal design is required to confirm the potential diagnostic efficacy of IL-10.

Fishmeal supplementation during ovine pregnancy and lactation protects against maternal stress-induced programming of the offspring immune system.

BACKGROUND: Prenatally stressed offspring exhibit increased susceptibility to inflammatory disorders due to in utero programming. Research into the effects of n-3 PUFAs shows promising results for the treatment and prevention of these disorders. The purpose of this study was to investigate whether maternal fishmeal supplementation during pregnancy and lactation protects against programming of the offspring's immune response following simulated maternal infection. METHODS: In order to accomplish this, 53 ewes were fed a diet supplemented with fishmeal (FM; rich in n-3 PUFA) or soybean meal (SM; rich in n-6 PUFAs) from day 100 of gestation (gd 100) through lactation. On gd135, half the ewes from each dietary group were challenged with either 1.2 µg/kg Escherichia coli lipopolysaccharide (LPS) endotoxin to simulate a bacterial infection, or saline as the control. At 4.5 months of age the offspring's dermal immune response was assessed by cutaneous hypersensitivity testing with ovalbumin (OVA) and candida albicans (CAA) 21 days after sensitization. Skinfold measurements were taken and serum blood samples were also collected to assess the primary and secondary antibody immune response. RESULTS: Offspring born to SM + LPS mothers had a significantly greater change in skinfold thickness in response to both antigens as well as a greater secondary antibody response to OVA compared to all treatments. CONCLUSIONS: Supplementation during pregnancy with FM appears to protect against adverse fetal programming that may occur during maternal infection and this may reduce the risk of atopic disease later in life.

Comparative studies on the metabolism of linoleic acid by rumen bacteria, protozoa, and their mixture in vitro.

Linoleic acid was differentially catabolized by the various rumen microbial fractions, such as rumen bacteria (B), protozoa (P), and their mixture (BP). The predominant isomer of conjugated linoleic acids (CLA) synthesized by B, P, and BP from linoleic acid was 9c11t-CLA. The formation of 9c11t-CLA was higher (P < 0.05) in P suspension (53.6 µg/mg microbial nitrogen) compared with B (38.3 µg/mg microbial nitrogen) and BP (28.8 µg/mg microbial nitrogen) suspensions by 12 h of incubation. The second most abundant CLA isomer was 10t12c. The accumulation of 10t12c-CLA in BP suspension was 2.3 times lower (P < 0.05) than that in B suspension (84.8 µg/mg microbial nitrogen) by 12 h of incubation. The accumulation of 10t-18:1 in BP suspension during 6- and 12-h incubation periods were not different (P > 0.05) than that in B suspension (6.8 and 14.0 µg/mg microbial nitrogen, respectively). However, the accumulation of 11t-18:1 in BP suspension at 6- and 12-h incubations were 2.7 and 3.3 times higher (P < 0.05), respectively, than that in B suspension. There were no significant accumulations of 11t-18:1, 10t-18:1, and 18:0 in P suspension throughout the incubation period. It was concluded that B, P, and BP metabolized linoleic acid to different isomers of CLA, whereas B, including BP, was only capable of biohydrogenating the CLA isomers to 18:0 by the reduction of 18:1 isomers. P was incapable of biohydrogenating LA, but its association with B in the BP suspension altered the biohydrogenation of LA significantly compared with B alone.

Effect of subacute ruminal acidosis on milk fat concentration, yield and fatty acid profile of dairy cows receiving soybean oil.

The objective of this study was to investigate the effect of ruminal infusion of soybean oil (SBO) with either a moderate- or high-forage diet on fat concentration, yield and composition in milk from dairy cows. Six rumen-fistulated Holstein dairy cows (639+/-51 kg body weight, 140+/-59 days in milk) were used in the study. Cows were randomly assigned to one of two dietary treatments, a high forage:concentrate (HFC, 74:26) or a moderate forage:concentrate (MFC, 56:44) total mixed ration. Cows were fed at 08.00 and 13.00 h and pulse-dosed ruminally at 13.00 h over a 10-min duration with 2% of diet dry matter of SBO. Ruminal pH was recorded continuously. Cows receiving the MFC treatment had lower daily mean ruminal pH and ruminal pH was below 6.0 for a longer duration compared with the HFC treatment (640 vs. 262 min/d, P<0.05). Cows receiving the MFC treatment had a greater reduction (diet by week interaction, P<0.05) in milk fat concentration and yield than cows receiving the HFC treatment (42 vs. 22% and 45 vs. 21%, respectively). Additionally, cows receiving the MFC diet had a greater reduction in milk fat concentration (g/100 g FA) of FA C16 (17 vs. 9%), trans-10 18:1 (159 vs. 21%) and trans-9, cis-11 conjugated linoleic acid (121 vs. 55%) (P<0.05) compared with cows receiving the HFC diet. This study demonstrated that cows fed the MFC diet had lower ruminal pH and showed a greater rate of milk fat depression when infused with SBO.

Microbial fatty acid conversion within the rumen and the subsequent utilization of these fatty acids to improve the healthfulness of ruminant food products.

Consumers are aware of foods containing microcomponents that may have positive effects on health maintenance and disease prevention. In ruminant milk, meat, and milk products; these functional food components include eicosapentaenoic acid (20:5n3), docosahexaenoic acid (22:6n3), 9c11t-conjugated linoleic acid, and vaccenic acid (11t-18:1). Modifying ruminal microbial metabolism of fatty acid in rumen through animal diet formulation is an effective way to enhance these functional fatty acids in ruminant-derived food products. However, it requires an understanding of the interrelationship between supply of lipid through the diet and rumen fermentation. Lipids in ruminant diets undergo extensive hydrolysis and biohydrogenation in the rumen. Apparent transfer efficiency of eicosapentaenoic acid and docosahexaenoic acid from feed to milk is very low (1.9 to 3.3%), which is, to a large extent, related to their extensive biohydrogenation in the rumen. Therefore, feeding a rumen-protected supplement containing eicosapentaenoic acid and docosahexaenoic acid, can be used to bypass the rumen. Ruminant-derived foods also contain different types of conjugated linoleic acid isomers, which are intermediates of rumen biohydrogenation of linoleic acid (9c12c-18:2). The predominant isomer of conjugated linoleic acid is 9c11t, which has numerous health benefits in animal models. The concentration of conjugated linoleic acid in ruminant-derived food products can be significantly enhanced through animal diet modification. We conclude that most current functional food products from ruminants have potential for their health-supporting properties, and for this market to succeed, an evidence-based approach should be developed in humans.

Fatty acid profile of bovine milk naturally enhanced with docosahexaenoic acid.

Recent studies have shown that the fatty acid profile of dietary lipid has the potential for improving the health of consumers. The present study was conducted to determine the fatty acid composition of commercial milks, namely, Dairy-Oh! Homo-Milk (DOHM), which is naturally enhanced with docosahexaenoic acid (DHA), or regular Homo-Milk (HM). The milk was collected from local supermarkets. The most abundant saturated fatty acids in the milk were butyric (C4:0), lauric (C12:0), myristic (C14:0), palmitic (C16:0), and stearic (C18:0) acids. Among unsaturated fatty acids, oleic acid (cis-9-C18:1) was also considerably high (502.7 mg/100 mL of milk). The concentration of total trans-18:1 was higher (P < 0.05) in DOHM than in HM (134.7 vs 107.0 mg/100 mL of milk, respectively), whereas total cis-18:1 was higher (P < 0.05) in HM than in DOHM (566.4 vs 508.4 mg/100 mL of milk, respectively). The concentration of DHA was 24.0 times higher (P < 0.05) in DOHM than in HM. DOHM contained 2.8 times higher (P < 0.05) eicosapentaenoic acid (EPA) compared to HM. Milk fat from DOHM contained a greater concentration of cis-9,trans-11 conjugated linoleic acid (CLA, 16.4 vs 11.6 mg/100 mL of milk, DOHM vs HM, respectively). The total omega-3 polyunsaturated fatty acids content was 2.23 times greater (P < 0.05) in DOHM compared with HM, due to an increase in C18:3n-3, EPA, and DHA. The result of the milk fatty acid analyses indicates that milk fat from DOHM had increased contents of EPA, DHA, and cis-9,trans-11 CLA, which could have a more favorable impact on diet composition and healthfulness.

Studies on the production of conjugated linoleic acid from linoleic and vaccenic acids by mixed rumen protozoa.

The present study was designed to investigate the capability of mixed rumen protozoa to synthesize conjugated linoleic acid (CLA) from linoleic (LA) and vaccenic acids (VA). Rumen contents were collected from fistulated cows. The protozoal fraction was separated and washed several times with MB9 buffer and then resuspended in autoclaved rumen fluid. The suspensions were anaerobically incubated up to 18 h at 38.5 degrees C with substrates in the presence (P-AB) or the absence of antibacterial-agents (P-No-AB). Neither P-AB nor P-No-AB suspensions were capable of producing CLA from VA (11t-18:1). Linoleic acid was catabolized by P-No-AB to a greater extent than P-AB. Different isomers of CLA were synthesized by P-AB from LA. The 9c11t-CLA was predominant. Thirty seven percent of the maximum accumulated 9c11t-CLA was found in the P-AB suspension as early as 0.1 h into the incubation period. Accumulation of 10t12c-CLA in P-AB suspension was approximately 10.0 times lower than that of 9c11t-CLA. There were no significant productions of VA, 10t-18:1, and 18:0 in P-AB compared with the control, indicating that rumen protozoa have no ability to biohydrogenate CLA isomers. On the other hand, the concentrations of 10t-18:1, VA, and 18:0 in P-No-AB were greater (P < 0.05) compared with those in P-AB, indicating the role of symbiotic bacteria associated with P-No-AB in biohydrogenating CLA isomers. We concluded that mixed rumen protozoa are capable of synthesizing CLA from LA through isomerization reactions. However, they are incapable of metabolizing CLA further. They are also incapable of vaccenic acid biohydrogenation and/or desaturation.

Fatty acid composition of yak (Bos grunniens) cheese including conjugated linoleic acid and trans-18:1 fatty acids.

The esterified fatty acid composition of cheese (YC) from yak ( Bos grunniens), reared in the highlands of the Nepalese Himalayas, was studied using capillary gas-liquid chromatography and compared with that of dairy cow Cheddar cheese (DC) purchased in a local market. The YC was collected from Dolakha, Nepal. The YC had a lower (P<0.001) myristic acid (C14:0; 6.7 vs 10.3%, YC vs DC, respectively) and palmitic acid content (C16:0; 23.3 vs 29.2%, YC vs DC, respectively) compared to DC. The YC had a lower (P<0.01) total medium-chain saturated fatty acids (C10:0-C16:0) content compared to DC (36.7 vs 47.3%, YC vs DC, respectively). On the other hand, the YC had a 24.8% higher (P<0.01) level of total long-chain saturated fatty acids (C17:0-C26:0) and a 3.2 times higher (P<0.001) content of total n-3 PUFA than DC. The ratio of n-3 PUFA to n-6 PUFA in YC was 0.87 compared to 0.20 in DC. YC had a 2.8 times higher (P<0.001) total trans-18:1 (9.18 vs 3.31%, YC vs DC, respectively) content. The percentage of vaccenic acid ( trans-11-C18:1) in YC was 4.6 times higher (6.23 vs 1.35% of total fatty acids, YC vs DC, respectively) than in DC. Vaccenic acid constituted 67.9% of total trans-C18:1 in YC. The Delta9-desaturase index for YC was lower than that of DC. The total conjugated linoleic acid (CLA) content in YC was 2.3% of total fatty acids compared to 0.57% in DC. The cis-9, trans-11 CLA isomer in YC constituted 88.5% of the total CLA. The results suggest that cheese from yak, grazed on Himalayan alpine pastures, may have a more healthful fatty acid composition compared to cheese manufactured from dairy cattle fed grain-based diets.

Conjugated linoleic acid increases skeletal muscle ceramide content and decreases insulin sensitivity in overweight, non-diabetic humans.

The effect of conjugated linoleic acid (CLA), a popular weight-loss supplement, on insulin sensitivity in humans is controversial and has not been extensively studied. To date no studies have examined the effects of CLA supplementation on human skeletal muscle metabolism or lipid content. It is also unknown whether CLA accumulates in human skeletal muscle with supplementation. In the present study, 9 overweight, non-diabetic individuals received 4 g/d of mixed CLA isomers in the form of 1 g supplements, for 12 weeks. CLA isomers significantly increased in both plasma and skeletal muscle following supplementation. Skeletal muscle ceramide content was also significantly increased, although there was no consistent change in muscle diacylglycerol or triacylglycerol content. Insulin sensitivity was significantly decreased (p

Method for determination of histidine in tissues by isocratic high-performance liquid chromatography and its application to the measurement of histidinol dehydrogenase activity in six cattle organs.

A selective and simple HPLC procedure has been developed to determine histidine (His) and histidinol (HDL) in liver supernate. The separation was performed on a column, Mightysil RP-18 GP. The eluted analytes were measured with UV detection without derivatization which provided detection limits of 1.1 and 2.0 microM for His and HDL (S/N ratio, 3:1), respectively. Recovery of the analytes added to liver sample was 98.3-101.6% within a 1-day study and 95.7-98.6% on different day (6 days) studies. The apparent histidinol dehydrogenase activities (nmol/g wet tissue) at pH 8, 9, 10, 11, and 12 were 38.6, 50.4, 160.3, 274.3, and 185.6 for liver; 90.6, 132.2, 30.7, 22.1, and 6.76 for kidney; 0.0, 0.0, 38.2, 20.1, and 12.9 for pancreas; 0.0, 0.0, 0.0, 14.7, and 6.8 for spleen; 0.0, 0.0, 4.2, 6.8, and 0.0 for muscle; and 0.0, 0.0, 4.9, 1.8, and 0.0 for small intestine, respectively. On the basis of optimum pH values, histidinol dehydrogenase activity in the organs was in the following order: liver>kidney>pancreas>spleen>muscle>small intestine.