We investigated the effects of dietary folic acid and age, on immunological parameters in laying hens challenged with LPS. 48 Shaver White hens at different ages were fed 2 wheat-soybean based diets with 0 or 4 mg supplemental FA per kg of diet for 8 wk. 6 hens from each dietary treatment were injected with 8 mg/kg body weight of LPS or saline and observed for 4 h. A few diet X challenge interaction were observed in young hens, whereas FA and LPS independently influenced a number of parameters. In older hens, there were very few diet X challenge interactions as well as effects of FA, whereas LPS affected several parameters. In conclusion, dietary FA influenced some immune responses in younger hens, but no such effects were observed in older hens. FA may modulate immune responses in laying hens under acute LPS challenge which could be tissue and age dependent.

Background: There has been a concern regarding the masking of vitamin B-12 deficiency in the post-fortification period (after January 1, 1998). Objective: The objective of this study was to investigate the potential masking of vitamin B-12 deficiency by comparing the proportion of individuals with low serum B-12 without macrocytosis between pre- and post-fortification periods using data from the National Health and Nutrition Examination Survey (NHANES). Methods: The study included 7242 individuals from NHANES III (pre-fortification group) and combined NHANES 1999-2000, 2001-2002, 2003-2004, 2005-2006 (post-fortification group). Vitamin B-12 deficiency and macrocytosis were defined as having <148 pmol/L of serum vitamin B-12 and mean corpuscular volume (MCV) of>98 fL, respectively. A multivariate logistic regression was performed to estimate the likelihood of being low serum B-12 without macrocytosis in the post-fortification period in relation to the pre-fortification period. Results: Between pre- and post-fortification periods, there was no significant difference in the proportion of individuals with low serum vitamin B-12 without macrocytosis. However, odds of having low serum vitamin B-12 without macrocytosis in the post fortification era increased in men (OR=2.65, 1.24-5.65), non-Hispanic blacks (OR=3.12, 1.04-9.35), Non-smokers (OR=4.63, 1.90-11.27), and those aged 55 and older (OR=2.183, 1.01-4.74) compared their respective counterparts in the pre-fortification period. Conclusions: No significant difference in the proportion of individuals with vitamin B-12 deficiency without macrocytosiss was seen between the pre and post-fortification periods suggesting no making of vitamin B-12 deficiency. In the post-fortification period, serum folate was found to be a predictor of the masking effect. The impact of increased folic acid intake in the post-fortification period needs to be evaluated on a periodic basis especially, in non-target population.

Hyperhomocysteinemia, a condition of elevated blood homocysteine level, is an independent risk factor for cardiovascular diseases. Folic acid can effectively reduce blood homocysteine levels. Recent studies have shown that hyperhomocysteinemia is also associated with liver disorders. However, the underlying mechanisms remain unclear. The general objective of my study was to investigate the biochemical and molecular mechanisms of homocysteine-induced liver injury and abnormal lipid metabolism. Hyperhomocysteinemia was induced in Sprague-Dawley rats by feeding a high-methionine diet for 4 weeks. An elevation of serum aminotransferases activities (indicator for liver injury) and an increase in hepatic lipid peroxidation were observed in hyperhomocysteinemic rats. Hyperhomocysteinemia-induced superoxide anion production led to oxidative stress in the liver. Reduction of oxidative stress by inhibiting superoxide anion production ameliorated hyperhomocysteinemia-induced liver injury. A significant elevation of hepatic and serum cholesterol concentrations in hyperhomocysteinemic rats was observed, exclusively due to increased expression of HMG-CoA reductase in hepatocytes. The molecular mechanisms of homocysteine-induced adverse effects were further investigated in isolated rat hepatocytes and in human hepatoma cells (HepG2). Hcy stimulated HMG-CoA reductase expression in hepatocytes via activation of transcription factors, namely, sterol regulatory element-binding protein-2 (SREBP-2), cAMP response element binding protein (CREB) and nuclear factor Y (NF-Y). Activation of these 3 transcription factors was detected in hyperhomocysteinemic rat liver and in homocysteine-treated hepatocytes. Pretreatment of hepatocytes with inhibitors for individual transcription factors effectively attenuated Hcy-induced HMG-CoA reductase mRNA expression. Supplementation of folic acid in diet significantly reduced serum homocysteine level and effectively inhibited hyperhomocysteinemia-induced superoxide anion production, resulting in amelioration of oxidative stress-mediated liver injury in hyperhomocysteinemic rats. These results reflected a protective role of folic acid in hyperhomocysteinemia-induced liver injury. In conclusion, the present study demonstrates that (1) hyperhomocysteinemia can cause oxidative stress and liver injury; (2) homocysteine stimulates cholesterol biosynthesis in hepatocytes via transcriptional regulation of HMG-CoA reductase expression; (3) supplementation of folic acid offers a hepatoprotective effect during hyperhomocysteinemia. Oxidative stress and accumulation of cholesterol in the liver contribute to liver injury associated with hyperhomocysteinemia. The role of folic acid in maintaining good health may extend beyond the cardiovascular system to encompass hyperhomocysteinemia-associated liver disorders.

The enrichment of egg with folate is a viable option for supplying the general population of a food product rich in natural folates. However, attempts to increase the concentration of folate in egg beyond the achieved level of enrichment had been unsuccessful because egg folate reached a maximum plateau when folic acid (FA) was increased in the diet. Thus, experiments were conducted to determine the factors regulating the deposition of dietary FA into the eggs. In the first study, the effect of feeding equimolar intake of FA and 5-methyltetrahydrofolate (5-methylTHF), the biologically active form of folate; on egg folate concentrations, indices of folate status, and activities of folate-dependent enzymes was evaluated. Folic acid and 5-methylTHF demonstrated equivalent effects in enhancing the egg folate concentrations and improving the indices of folate status in the laying hen. The activities of folate-dependent enzymes were similar between the two forms of folate except for hepatic dihydrofolate reductase (DHFR) activity which increased in FA-fed birds compared to 5-methylTHF-fed birds. However, this demonstrated the ability of the laying hen to metabolically convert FA into its biologically active forms. Therefore, the influence of intestinal FA absorption in the regulation of FA deposition in the egg was subsequently evaluated. Using the in vitro everted intestinal sac technique, FA was absorbed in all regions of the intestine. Absorption was maximum at acidic pH 6.0, and increased in the duodenum and jejunum compared to the ileum and cecum. The rate of FA absorption in the jejunum diminished at higher FA concentrations. Therefore, further study was conducted to determine the regulation of FA absorption when levels of FA in the laying hen diet are increased. Supplementation of increased FA levels resulted to a down-regulation of FA absorption in the duodenum, but not in the jejunum of the laying hen. This down-regulation was not associated to a decreased mRNA gene expression of the duodenal folate transporters. Overall, decreased intestinal rate of FA absorption possibly associated to a post-transcriptional or translational regulation of specific folate transporters in the intestine of the laying hen may contribute to the saturation in the egg folate concentration.

We investigated the effects of dietary folic acid and age, on immunological parameters in laying hens challenged with LPS. 48 Shaver White hens at different ages were fed 2 wheat-soybean based diets with 0 or 4 mg supplemental FA per kg of diet for 8 wk. 6 hens from each dietary treatment were injected with 8 mg/kg body weight of LPS or saline and observed for 4 h. A few diet X challenge interaction were observed in young hens, whereas FA and LPS independently influenced a number of parameters. In older hens, there were very few diet X challenge interactions as well as effects of FA, whereas LPS affected several parameters. In conclusion, dietary FA influenced some immune responses in younger hens, but no such effects were observed in older hens. FA may modulate immune responses in laying hens under acute LPS challenge which could be tissue and age dependent.

Methylenetetrahydrofolate reductase (MTHFR), an important enzyme in folate metabolism, mediates the conversion of 5,10-methylenetetrahydrofolate to 5-methyltetrahydrofolate which serves as the carbon donor for the conversion of homocysteine to methionine. It is also inhibited by S-adenosylmethionine which has shown to be actively demethylated to form S-adenosylhomocysteine, which is hydrolysed to homocysteine. MTHFR deficiency exhibits well-documented clinical and biochemical symptoms. The human MTHFR cDNA was isolated by Goyette et al (1994), and fifteen mutations have been identified at this locus. / An animal model would prove to be useful for designing therapeutic approaches for understanding the pathogenesis of this genetic disease at the molecular level. The mouse MTHFR gene and cDNA have been isolated and partially characterized. Four genomic clones were isolated by library screening. One of these clones (clone 3) contained the 5$ sp prime$ end of the gene and was completely characterized. The clone was shown to have no rearrangements and is to be used to design targeting vectors for 'knockout mice' and mice carrying a common mutation which has been postulated to be a genetic risk factor for cardiovascular disease. The other three clones contain the remaining 3$ sp prime$ portion of the gene. The coding portion has approximately 90% homology with the human cDNA and also shows a similar gene structure. / A 2.2 Kb mouse MTHFR cDNA was isolated by library screening and was found to contain a 320 base pair extension at the 5$ sp prime$ end which has not been found in the human cDNA. The cDNA contains exons -1 -3, but also contains two possibly unspliced introns. A portion of this cDNA can however still be used to rescreen libraries to isolate a full length cDNA. The above research is the first genetic data on the mouse MTHFR gene and provides the basis for future research involving mouse models of MTHFR deficiency.

We have studied the effect of S-adenosylmethionine (SAM) on tail flick latency in the rat. We also studied the effect of methionine the immediate precursor of SAM. Administration of methionine to the rat increases brain SAM, but little is known about its behavioral effects. Long-Evans rats were given SAM and methionine orally at different doses and tail-flick latency was measured at various times. Both methionine and SAM increased tail-flick latency, but methionine did so at a lower dose. A biochemical study showed that methionine was more effective than SAM in raising brain SAM probably because it is transported better into brain. The biochemical measurements were not consistent with the idea that the effects of SAM and methionine were mediated by an increase in brain 5-HT. / Folate deficiency can lower brain SAM levels and cause depression. Thus, methionine, which raises brain SAM, may overcome the effects of folate deficiency. Seven day food records were done by 26 psychiatric outpatients who were stable on lithium treatment. Eight patients had mean daily folate intakes below those recommended. Some of those with low folate intake had high methionine intake consistent with the idea that methionine could substitute metabolically for folate deficiency. Daily methionine intakes ranged from 13 to 304% of the recommended intake. As methionine had behavioral effects in the rat at doses much less than the daily dietary intake this raises the question of whether varying daily intakes of methionine in humans have behavioral implications. (Abstract shortened by UMI.)

Micronutrient poor diets around the globe and in particular in the developing world cause deficiencies in iron and folic acid. This may be rectified by the incorporation of these micronutrients into currently running salt iodization processes. The objective of this project was to develop folic acid and iodine spray solutions to be ready for pilot scale testing and to investigate the stability of triple fortified salt containing iodine, folic acid and microencapsulated ferrous fumarate. The optimal spray solutions were buffered to pH 9 with a carbonate/bicarbonate buffer to stabilize folic acid and contained 1%-2% w/v folic acid and 1%-3% w/v iodine (as KIO3). They remained in solution and retained ≥80% of both micronutrients after 5 months of storage at 25ºC and 45ºC. Double fortified salt produced using these spray solutions retained 100% of both folic acid and iodine over a 5 month period when stored at ambient conditions. Unfortunately triple fortified salt did not sufficiently retain the micronutrients due to excess moisture absorption and inadequate encapsulation of iron.

Micronutrient poor diets around the globe and in particular in the developing world cause deficiencies in iron and folic acid. This may be rectified by the incorporation of these micronutrients into currently running salt iodization processes. The objective of this project was to develop folic acid and iodine spray solutions to be ready for pilot scale testing and to investigate the stability of triple fortified salt containing iodine, folic acid and microencapsulated ferrous fumarate. The optimal spray solutions were buffered to pH 9 with a carbonate/bicarbonate buffer to stabilize folic acid and contained 1%-2% w/v folic acid and 1%-3% w/v iodine (as KIO3). They remained in solution and retained ≥80% of both micronutrients after 5 months of storage at 25ºC and 45ºC. Double fortified salt produced using these spray solutions retained 100% of both folic acid and iodine over a 5 month period when stored at ambient conditions. Unfortunately triple fortified salt did not sufficiently retain the micronutrients due to excess moisture absorption and inadequate encapsulation of iron.

Thesis (M.P.H.)--Georgia State University, 2007. / Title from file title page. Russ Toal, committee chair; Catherine McCarroll, Joseph Mulinare, committee members. Electronic text (83 p. : ill.) : digital, PDF file. Description based on contents viewed Jan 8, 2008. Includes bibliographical references (p. 79-83).