No description available.

Oxidative stress has been associated with a wide range of diseases, including cardiovascular diseases, Alzheimer's disease, atherosclerosis, Parkinson's disease and cancer. It also plays a role in the ageing process. Hyperhomocysteimia is commonly found to be associated with these diseases. The hyperhomocysteimia is a result of a deficiency in both folate and cobalamin Folate is known to reduce Hey and protect cells from apoptosis, but there are no studies investigating the impact of cobalamin on apoptosis induced by oxidative stress or the mechanism(s) of the protection. The aims of the research are to investigate the protective role of cobalamin and the possible mechanism(s) for this protection. It also examines the protective role of novel cobalamin and investigates their superior protection. The methods used in this research for apoptosis detection we used caspase-3 and the annexin-V, while for necrosis we used PI staining, where cell viability were detected using MTS assay. We also measured the generation of superoxide by Lucigenin-enhanced chemiluminescence and reactive oxygene species by using the redox active prob DCFH-DA. Moreover, the intracellular proteins were measured via staining with specific fluorescent-conjugated antibodies were detected using flowcytometry. Our result demonstrated that 25|iM of cobalamin protects cells from apoptosis. The protection by cobalamin was associated with induction of iHsp72 and iHO-1, and these are shown to be essential for the protection. Furthermore, our research demonstrated a novel mechanism of cobalamin-apoptosis protection involving induction of NfkB, ERK1/2 and AKT signal transduction pathways. In order to protect cells from apoptosis induced by oxidative stress, cobalamin induces the pNfkB which in turn regulate the iNOS and HO-1 induction. Cobalamin also induces thepERK1/2 which regulates the induction of Hps72 and Nrf2. And finally, pAKT induced by cobalamin which regulate the Nrf2 and HO-1 induction. The inhibition of any of theses pathways leads to loss the protection. The GSCbl and NACCbl provide a superior protection against oxidative stress, this protection involved induction of the signal transduction pathways and Hsps. To conclude; cobalamin provides protection against cells death induced by oxidative stress. Cobalamin achieves this by multiple pathways which include direct antioxidant stimulation and induction of signal transduction pathways. Different cobalamin derivatives have superior protections. These finding are a useful pharmaceutical tool in the treatment of the oxidative stress related diseases.

No description available.

No description available.

abstract: According to a 2016 census, eight million adults conform to a vegetarian diet within the United States, and about 50% of these adults follow a vegan diet. The census determined that plant-based diets are quickly growing in popularity particularly in young adults between the ages of 18 to 34 years. Many Americans are aware of the health benefits of a plant-based diet, however, the dietary risks associated with these diets are not well emphasized. Health concerns such as vitamin deficiencies and altered metabolism are heightened in vegetarian populations. One Particular nutrient that is commonly lacking in the vegetarian diet is vitamin B12. Vitamin B12 is found mainly in animal-derived food sources such as meat, poultry, fish, dairy, and eggs. Although some vegetarians, called lacto-ovo vegetarians, consume dairy and eggs, vegans do not consume any animal products at all. Vitamin B12 deficiency can have devastating consequences on the human body due to its role as a methylation cofactor. Metabolism, DNA replication, and cancer formation all involve methylation processes. This cross-sectional, differential study aimed to further understand the relationship between vegetarianism, vitamin B12 status, and methylation capacity in healthy adults. A group of 34 healthy adults (18 vegetarians and 16 omnivores) was recruited to analyze serum B12, homocysteine, methylmalonic acid, serum total folate, and transcobalamin II status. It was hypothesized that (1) vegetarians would have a lower vitamin B12 status, and thus, a lower methylation capacity than omnivores and that (2) low vitamin B12 status would be correlated with low methylation capacity. The data show that vegetarians did not have significantly lower vitamin B12 methylation capacity status than omnivores. Nor was vitamin B12 status correlated with methylation capacity. However, the data revealed that diet quality had a positive influence on folate status. There was also a statistical trend (p=0.08) for homocysteine reduction in participants consuming high-quality diets. The data herein suggest that methylation capacity may be impacted by the quality of diet rather than the type of diet. / Dissertation/Thesis / Masters Thesis Nutrition 2019

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Non-invasive markers that are clinically useful for the diagnosis and monitoring of canine chronic enteropathies are scarce. The first aim of this study was to investigate the prevalence of cobalamin deficiency on a cellular level in dogs with chronic gastrointestinal disease by measuring serum methylmalonic acid (MMA) concentrations. Hypocobalaminemia has been associated with a negative outcome in dogs with chronic enteropathies, but the prevalence of cellular cobalamin deficiency is unknown. The second aim of this study was to determine the utility of fecal and urinary concentrations of N-methylhistamine (NMH) as a marker of gastrointestinal inflammation and disease activity in dogs with chronic enteropathies. Serum MMA concentrations were measured in healthy control dogs to establish a reference interval, which was calculated to be 415-1,193 nmol/L. Measurement of MMA concentrations in 555 serum samples from dogs with varying cobalamin concentrations showed a significant increase (p<0.05) in dogs with hypocobalaminemia. In a prospective group of 56 dogs with chronic enteropathies, 36% had decreased serum cobalamin concentrations, five of which (9% of 56 dogs) had increased serum MMA concentrations. We conclude that hypocobalaminemia is commonly seen in dogs with chronic gastrointestinal disease, but does not always appear to be associated with cellular cobalamin deficiency. In 47 dogs with chronic enteropathies, fecal and urinary NMH concentrations were increased in 21% and 27%, respectively, indicating that mast cell degranulation plays a role in a subset of dogs with chronic enteropathies. However fecal and urinary NMH concentrations did not correlate with each other, or with the clinical activity index. Urinary NMH concentrations correlated significantly with serum CRP concentrations, and were also significantly associated with severity of duodenal mucosal inflammation (p=0.008). The lack of correlation with the clinical activity index suggests that degranulation of mast cells only plays a role in some dogs with chronic enteropathies. Also, these results suggest that NMH alone may not be a good marker for clinical disease activity in dogs with chronic enteropathies. Due to its linear association with serum CRP and severity of mucosal inflammation, urinary NMH concentrations may be a better marker of intestinal inflammation than fecal NMH concentrations.

abstract: Vitamin B12, found only in animal products, is a water-soluble vitamin important for DNA methylation, purine and pyrimidine synthesis, and the myelination of nerves. Symptoms of vitamin B12 deficiency include anemia, gait disturbances, altered vibration proprioception, impaired vision, psychosis, depression, dementia-like illness, and ultimately death. Because vegetarians and vegans consume fewer animal products in their diet than omnivores, they are inherently more at risk for developing these symptoms of vitamin B12 deficiency. Thus, the purpose of this study is to examine the correlation between nervous system markers (balance, dexterity, and vibration sensitivity) and markers of vitamin B12 nutriture (serum B12 and serum holo-transcobalamin II) in a cross-sectional study (n=38). In addition, the impact of daily oral vitamin B12 supplementation on these markers in an 8-week randomized controlled trial was also examined (n=18). The results of the cross-sectional study revealed a moderate correlation (R=-0.351, p=0.031) between serum B12 and left-hand functional dexterity. The results of the intervention study revealed no significant time*group interactions for markers of nervous system functions and biochemical values (after the removal of outliers). In addition, the time*group interaction appeared to be larger for those individuals with a baseline serum B12 of less than 303 pmol/L. These results suggest that vitamin B12 supplementation may have a more pronounced effect on those individuals who are in a state of vitamin B12 depletion (<303 pmol/L serum concentration). In addition, the results also suggest that 8 weeks of oral supplementation is not a long enough period to create significant clinical change, and it is likely that improvements in neurological measures would require long-term supplementation. / Dissertation/Thesis / Doctoral Dissertation Physical Activity, Nutrition and Wellness 2016

Einleitung: Vitamin B12 hat wichtige Funktionen im Energiestoffwechsel sowie bei der Erythropoese. Beide Funktionskreise werden bei Hochleistungskühen besonders beansprucht und können bei Belastungen und ungenügender Vitamin-B12-Versorgung Ausgangspunkt für klinische Störungen werden. Ziele der Untersuchungen: In den vorliegenden Studien wurde der Fragestellung nachgegangen, wie sich die Vitamin-B12-Konzentration im Blutserum von Milchkühen in der Frühlaktation verhält und welche Zusammenhänge zu Stoffwechselparametern, dem Erythrogramm sowie dem Gesundheitsstatus der Kühe bestehen. Des Weiteren wurde geprüft, inwieweit der postpartale Stoffwechsel und der Gesundheitsstatus durch orale Vitamin-B12-Substitutionen stabilisiert werden können. Material und Methoden: Die Untersuchungen zur Vitamin-B12-Statuserhebung erfolgten an 157 Kühen der Rasse Holstein Friesian. Blutproben zur Stoffwechselanalytik wurden 2-6 Tage p.p. sowie 4-5 Wochen p.p. entnommen. Parallel dazu wurden klinische Daten zu Leistung und Gesundheitsstatus bis 3 Monate p.p. erhoben. In einem zweiten Versuch wurden die Kühe in 2 Gruppen eingeteilt, wobei die Versuchsgruppe (65 Kühe) eine orale Vitamin-B12-Substitution in Höhe von 0,5 g Cyanocobalamin/Kuh/Tag 4-6 Wochen a.p. beginnend bis zur Kalbung erhielt. 71 Kühe, die das stallübliche Mineralfutter erhielten, dienten als Kontrollgruppe. Auch hier erfolgten die Blutkontrollen 2-6 Tage p.p. sowie 4-5 Wochen p.p. Es wurden die Milchleistung sowie auftretende Erkrankungen dokumentiert. Die Blutentnahme erfolgte aus der Vena caudalis mediana. Neben den hämatologischen Untersuchungen wurden folgende Parameter aus dem Serum bestimmt: Freie Fettsäuren (FFS), Betahydroxybutyrat (BHB), Glukose, Bilirubin, Cholesterol, Gamma-Glutamyltransferase (GGT), Creatinkinase (CK), Harnstoff, Calcium, Eisen, anorganisches Phosphat (Pi), Cyanocobalamin und Cobalt. Ergebnisse: Die Vitamin-B12-Konzentration zeigt eine signifikante Laktationsdynamik. Alle untersuchten Kühe hatten 4 Wochen p. p. gegenüber 2–6 Tage p. p. erniedrigte Vitamin-B12-Konzentrationen (p ≤ 0,05). Bei den p.p. kranken Kühen sank gegenüber den gesunden Kühen die Vitamin-B12-Konzentration weniger stark ab (p ≤ 0,05), d.h., höhere Vitamin-B12-Konzentrationen können auf klinische Probleme hinweisen. Gesunde sowie auch p.p. kranke Kühe wiesen 2–6 Tage p. p. höhere Werte für die Parameter Erythrozytenzahl, Hämatokrit und Hämoglobinkonzentration auf als 4 Wochen p. p. Die BHB-, FFS- und Bilirubin-Konzentrationen waren bei allen Kühen 2–6 Tage p. p. infolge der partusbedingten Lipolysesteigerung erhöht (p ≤ 0,05). Bei allen Kühen korrelierte die Aktivität der cholestaseanzeigenden GGT eng mit der Vitamin-B12-Konzentration (p ≤ 0,01). Aufgrund dieser engen Korrelation mit der GGT-Aktivität sowie der Bilirubinkonzentration kann Vitamin B12 bei einer Serumkonzentration ≥ 227 ng/l bei Kühen cholestatische Stoffwechselbelastungen anzeigen. Nach Vitamin-B12-Substitution blieben in der Versuchsgruppe 60 % und in der Kontrollgruppe 47,9 % der Kühe in der Frühlaktation gesund. In der Kontrollgruppe hatten 12,7 % eine Nachgeburtsverhaltung und 40,8 % eine Mastitis, in der Versuchsgruppe betrugen die Anteile 21,5 % sowie 26,2 %. Mit x̃ = 320 pg/ml war die Vitamin-B12-Konzentration 2-6 Tage p.p. in der Vitamin-B12-substituierten Gruppe gegenüber x̃ = 224 pg/ml in der Kontrollgruppe gesichert höher. Auch vier Wochen p.p. war die Differenz noch signifikant. Cobalt, die Parameter des Leber- und Energiestoffwechsels sowie Harnstoff und CK unterschieden sich zwischen der Vitamin-B12-substituierten Gruppe und der Kontrollgruppe nicht gesichert. Die Erythrozytenzahlen sowie die Hämoglobin-Konzentrationen waren in der Vitamin-B12-substituierten Gruppe gesichert höher. Der Milchfettgehalt (%) war bei den Vitamin-B12-substituierten Kühen gegenüber den Kontrollkühen signifikant erhöht (p = 0,022), die Milchleistung unterschied sich unwesentlich. Schlussfolgerungen: Signifikant höhere Vitamin-B12- und Hämoglobin-Konzentrationen, höhere Erythrozytenzahlen sowie geringere Morbidität sprechen für positive Effekte der Vitamin-B12-Substitution. Anhand der Parameter des Leber-Energiestoffwechsels sowie der Milchleistung ließ sich dies nicht bestätigen. Cholestase stört die Vitamin-B12-Bewertung im Blut bzw. ist bei der Interpretation zu beachten. / Introduction: Vitamin B12 has important functions in energy metabolism and erythropoiesis. Both functional groups are particularly stressed in high-yielding dairy cows and can be a starting point for clinical disorders under stress and insufficient vitamin B12 supply. Objective: The studies presented were designed to ascertain the characteristics of the serum vitamin B12 concentration of dairy cows in early lactation and to check the relations with metabolic parameters, the erythrogram as well as the health status of the cows. Furthermore, it was examined to what extent the postpartum metabolism can be stabilized by oral vitamin B12 substitutions. Material and methods: The investigations on vitamin B12 status survey were carried out on 157 cows of the Holstein Friesian breed. Blood samples were taken for metabolic analysis at 2-6 days p.p. and at 4-5 weeks p.p. In parallel, clinical findings on the milk yield and the health status were compiled up to 3 months p.p. In a second trial the cows were divided into 2 groups in which the experimental group (65 cows) received an oral vitamin B12 substitution in the amount of 0.5 g cyanocobalamin/cow/day starting 4-6 weeks a.p. up to calving. 71 cows, which recieved the common mineral feed, served as control group. Again the blood tests were performed at 2-6 days p.p. and at 4-5 weeks p.p. The milk yield and emerging diseases were documented. The blood samples werde taken from the Vena caudalis mediana. In addition to haematological investigations the following parameteres were measured: Non-esterified fatty acids (NEFA), beta-hydroxybutyrate (BHB), glucose, bilirubin, cholesterol, gamma-glutamyl transferase (GGT), creatinkinase (CK), urea, calcium, ferric, anorganic phosphor, cyanocobalamin and cobalt. Results: The vitamin B12 concentration shows a significant dynamic in lactation. All examined cows had decreased vitamin B12 concentrations at 4 weeks p. p. compared to 2-6 days p. p. (p ≤ 0.05). In the p.p. morbid cows the vitamin B12 concentration fell less than in the healthy cows (p ≤ 0.05), which means higher vitamin B12 concentrations may indicate clinical problems. Healthy as well as p.p. morbid cows showed higher values for the parameters erythrocyte count, hematocrit and hemoglobin concentration at 2-6 days p.p. than 4 weeks p.p. BHB, FFS, and bilirubin levels were increased in all cows at 2-6 days p.p. as a result of partus related rise in lipolysis (p ≤ 0.05). In all cows, the activity of the GGT, which indicates cholestasis, was closely correlated with the vitamin B12 concentration p ≤ 0.01). Because of this close correlation with GGT activity and bilirubin concentration, vitamin B12 may show cholestatic metabolic stress in cows at a serum concentration ≥ 227 ng/l. After vitamin B12 substitution 60 % of the cows in the experimental group and 47.9 % of the cows in the control group remained healthy during early lactation. In the control group 12.7 % had a Retentio secundinarum and 40.8 % had a mastitis, in the experimental group the proportions were 21.5 % and 26.2 %. At two to six days p.p. the vitamin B12 concentration in the vitamin B12 substituted group was significant higher (x̃ = 320 pg/ml) than in the control group (x̃ = 224 pg/ml). This difference was still significant at four weeks p.p. Cobalt, parameters of liver and energy metabolism as well as urea and CK did not differ significantly between the vitamin B12 substituted group and the control group. Erythrocyte counts and hemoglobin concentrations were significant higher in the vitamin B12 substituted group. Milk fat content (%) was significant higher in the vitamin B12 substituted group compared to the control group (p = 0.022), the milk yield did not differ significantly. Conclusions: Significant higher vitamin B12 and hemoglobin concentrations, higher erythrocyte counts as well as lower morbidity speak for positive effects of vitamin B12 substitution. Based on the parameters of hepatic and energy metabolism as well as milk yield this could not be confirmed. Cholestasis interferes with the evaluation of vitamin B12 in the blood respectively should be considered in the interpretation.

O aborto espontâneo recorrente (AER) é caracterizado pela ocorrência de três ou mais abortos consecutivos e acomete 2-4% das mulheres em idade fértil. A etiologia está associada a vários fatores de risco, tais como anomalias uterinas, aberrações cromossômicas, autoimunidade, trombofilias, elevação na concentração de homocisteína (tHcy), porém cerca de 40% dos casos permanece sem causa definida. O metabolismo de unidades de carbono desempenha papel fundamental na disponibilidade de folato na célula, sendo essencial para o desenvolvimento placentário e fetal. Deficiência de vitaminas que regulam este metabolismo, como o ácido fólico, e polimorfismos em genes que codificam enzimas relacionadas ao metabolismo de folato (MTHFR, RFC1, GCP2 e MTHFD1) podem levar à redução das concentrações desta vitamina e ao aumento das concentrações de tHcy. Objetivo foi avaliar a associação entre polimorfismos em genes relacionados ao metabolismo do folato (RFC1, GCP2, MTHFR e MTHFD1) e o risco de se ter AER, bem como avaliar a associação entre estes polimorfismos e as alterações nas concetranções de folato, cobalamina e homocisteína. Foram constituídos três grupos: AER primário: 117 mulheres com AER e nenhum feto viável; AER secundário: 139 mulheres com AER e pelo menos um feto viável; e Controle: 264 mulheres sem história de aborto espontâneo. Nenhuma das mulheres estava grávida no momento da coleta do sangue. Amostras de sangue foram obtidas para dosagens bioquímicas (folato, Cbl, tHcy, entre outras), imunológicas e extração de DNA genômico. As genotipagens foram feitas por PCR-RFLP ou PCR em tempo real. As concentrações séricas de folato e Cbl foram maiores no AER primário e secundário (p<0,05). A distribuição dos genótipos de todos os polimorfismos foi semelhante nos três grupos. O aumento nas concentrações de folato sérico (OR: 1,05, 95% IC: 1,03 - 1,07, p<0,001), Cbl (OR: 1,00, 95% IC: 1,00 - 1,00, p= 0,016), tHcy (OR: 1,03, 95% IC: 0,97 - 1,11, p= 0,033) e T4 (OR: 1,02, 95% IC: 1,00 - 1,03, p= 0,006) e a presença de FAN reagente (1:160) (OR: 2,90, 95% IC: 1,25 - 6,75, p= 0,013) foram considerados fatores de risco para aborto primário. Para o aborto secundário, foram considerados fatores de risco o aumento nas concentrações de folato sérico (OR: 1,04, 95% IC: 1,02 - 1,05, p<0,001), Cbl (OR: 1,00, 95% IC: 1,00 - 1,00, p= 0,019) e tHcy (OR: 1,05, 95% IC: 1,00 - 1,09, p= 0,039), maiores idades (OR: 1,02, 95% IC: 0,98 - 1,06, p= 0,031), hábito de fumar (OR: 2,54, 95% IC: 1,41 - 4,60, p= 0,002) e ter maior IMC (OR:1,42, 95% IC: 1,07 - 1,88, p= 0,015). Os polimorfismos estudados não foram associados ao maior risco de se ter AER, quando analisados isoladamente, e também não foram associados a alterações nas concentrações séricas de folato, Cbl e tHcy, com exceção do genótipo MTHFR 677TT, cujas portadoras apresentaram maior concentração de tHcy, quando comparadas com as portadoras de genótipos 677CC e 677CT nos três grupos. As variáveis concentrações de folato, Cbl, tHcy e T4 e presença de FAN reagente foram associadas ao maior risco de se ter aborto primário. As variáveis idade, IMC, tabagismo, concentrações de folato, Cbl e tHcy foram associadas ao maior risco de aborto secundário. / The recurrent spontaneous abortion (RSA) is characterized by the occurrence of three or more consecutive miscarriages and affects 2-4% of women of childbearing age. The etiology is associated with several risk factors such as uterine abnormalities, chromosomal aberrations, autoimmunity, thrombophilia, increased concentration of homocysteine (tHcy). About 40% of cases remains unknown cause. The units of carbon metabolism plays an essential role in the availability of the cell folate, is essential for the placental and fetal development. A deficiency of the vitamins that regulate this metabolism, like folic acid, and polymorphisms in genes encoding enzymes related to folate metabolism (MTHFR, RFC1, and GCP2 MTHFD1) may lead to decreased concentrations of this vitamin and increased concentrations of tHcy. Objective was to evaluate the association between polymorphisms in genes related to folate metabolism (RFC1, GCP2, MTHFD1 and MTHFR) and the risk of having AER, and to evaluate the association between these polymorphisms and changes in concetranções folate, cobalamin, and homocysteine. Three groups were divided: AER primary: 117 women with RSA and no viable fetus, AER secondary: 139 women with RSA and at least one viable fetus and Control: 264 women with no history of miscarriage. None of the women was pregnant at time of blood collection. Blood samples were taken for biochemical (folate, Cbl, tHcy, etc.), immunological and genomic DNA extraction. The genotyping were carried out by PCR-RFLP or real time PCR. Serum concentrations of folate and Cbl were higher in groups 1 and 2 (p <0.05). The distribution of genotypes of MTHFR c.677C> T, MTHFR c.1298A> C, MTHFD1 c.1958G> A, RFC1 c.80G>GCP2 A and c.1561C> T was similar among the three groups. The increased concentrations of serum folate (OR: 1.05, 95% CI: 1.03 - 1.07, p <0.001), Cbl (OR: 1.00, 95% CI: 1.00 to 1.00, p = 0.016), tHcy (OR: 1.03, 95% CI: 0.97 to 1.11, p = 0.033) and T4 (OR: 1.02, 95% CI: 1.00 to 1.03, p = 0.006) and the presence of ANA (1:160) (OR: 2.90, 95% CI: 1.25 - 6.75, p = 0.013) were considered risk factors primary for abortion. For secondary abortion, were considered risk factors increased the concentrations of serum folate (OR: 1.04, 95% CI: 1.02 - 1.05, p <0.001), cobalamin (OR: 1.00, 95 % CI: 1.00 to 1.00, p = 0.019) and tHcy (OR: 1.05, 95% CI: 1.00 to 1.09, p = 0.039), higher age (OR: 1.02, 95% CI: 0.98 to 1.06, p = 0.031), cigarette smoking (OR: 2.54, 95% CI: 1.41 to 4.60, p = 0.002) and had a higher BMI (OR : 1,42,95% CI: 1.07 to 1.88, p = 0.015). The studied polymorphisms were not associated with increased risk of having RSA when analyzed separately, and were not associated with changes in serum folate, Cbl and tHcy, with the exception of the MTHFR 677TT genotype, whose patients had a higher concentration of total tHcy compared with those with 677CC and 677CT genotypes in the three groups. The variable concentrations of folate, Cbl, tHcy, and T4, presence of ANA and have been associated with increased risk for miscarriage primary. The variables age, BMI, smoking, concentrations of folate, Cbl and tHcy were associated with increased risk of secondary miscarriage.