Vitamin B6 Dependence of One-Carbon Metabolism
Condition: Vitamin B6 Deficiency
MeSH terms: Vitamin B 6 Deficiency
Study type: Interventional

Summary: 
      Marginal vitamin B6 deficiency, which occurs commonly worldwide, leads to a cellular
      deficiency of the coenzyme pyridoxal phosphate (PLP). PLP is a coenzyme in several phases of
      one carbon (1C) metabolism, which is the array of reactions in which one carbon units are
      acquired and used in reactions including nucleotide synthesis, regeneration of methionine
      (Met) from homocysteine (Hcy), and methylation of many biological compounds. 1C metabolism is
      linked to the transsulfuration pathway in which Hcy undergoes PLP-dependent catabolism
      leading to cysteine, whose availability governs the formation of the antioxidant glutathione.
      Nutritional or genetic conditions that impair 1C metabolism are associated with elevation in
      plasma Hcy concentration and increased risk of vascular disease. It is believed that the
      metabolic effects of vitamin B6 deficiency will be most pronounced following protein intake
      when the vitamin B6-dependent pathways of amino acid metabolism experience the greatest
      substrate load. The human subjects protocols of this study consist of two distinct phases
      intended to extend our understanding of basic human 1C metabolism and the effects of marginal
      vitamin B6 deficiency under postprandial conditions. Phase 1 will investigate the effects of
      vitamin B6 nutrition on the PLP-dependent generation of 1C units by the glycine cleavage
      system and on the synthesis of glutathione. Phase 2 will investigate the dependence of
      methionine metabolism on vitamin B6 nutritional status, with particular emphasis on the
      recycling of Hcy to Met. Each phase of this study will involve 14 healthy, nutritionally
      adequate, young adults (7 male, 7 female) who will undergo metabolite profiling and kinetic
      analysis using intravenously infused stable isotopic tracers performed both before and after
      a ~4-week period of dietary vitamin B6 restriction. Subjects will be assigned to either Phase
      1 or Phase 2, which will be identical in design except for the tracers and analytical methods
      used. We hypothesize that vitamin B6 deficiency will yield reduction in postprandial rates of
      homocysteine remethylation, generation of 1C units from glycine, and synthesis of
      glutathione. The results of this study will aid in assessing the consequences of nutritional
      and genetic variables affecting human metabolism and will further our understanding of the
      relationships between vitamin B6 nutrition and disease.
    

Detailed description: 
      If you volunteer for this study, you will come to the General Clinical Research Center (GCRC)
      at Shands Hospital for a screening visit. There approximately 27.5 mL of blood (5 1/2
      teaspoons) will be taken from a vein in your arm. This will be used for tests to determine
      whether you are receiving adequate amounts of vitamin B6 and other vitamins, to analyze a
      part of the genetic information in certain blood cells that relates to how your body
      processes some nutrients, and to confirm your general health. If the vitamin B6 content of
      your blood is higher than the range needed for this study, which sometimes occurs from eating
      fortified foods, you might be asked to give another blood sample (2 teaspoons) after several
      weeks to have the screening test repeated. You will also be asked questions about your health
      and a detailed assessment of your regular diet will be carried out. Your general health will
      be confirmed by taking a medical history, physical exam, and standard blood tests including
      liver, kidney, bone marrow function, cholesterol and urine analysis. To take part in this
      study, you must be in good health and in adequate nutritional status for vitamin B6 and other
      vitamins.

      Because of risks to a developing fetus from inadequate vitamin B6 intake, a pregnancy test
      will be conducted with females three times during the study (during the screening visit and
      before each hospital admission) and women who participate in this study should use protection
      to prevent pregnancy during the study; if you become pregnant you will be released from the
      study.

      Once you have completed all screening procedures you will be asked to eat meals prepared by
      the GCRC for 2 days, you will then participate in the procedure to test how effectively
      vitamin B6 functions in helping your body use certain amino acids. This will involve giving
      you a solution of certain amino acids by a vein in your arm over the course of 9 hours.

      On the evening before this procedure, you will report to the GCRC at approximately 8:00 p.m.
      You will spend the night at the GCRC and will not be allowed to eat after 9:00 p.m. On the
      following morning, you will not be given breakfast. You will have an intravenous catheter
      placed into a vein in each arm, one for blood samples to be taken throughout the day, and one
      for the amino acid solution to be given. You will also be asked to breath (one breath)
      through a straw into a glass tube at different times, while the amino acid solution is being
      given. Before starting the amino acid solution, a blood sample (about 7 teaspoons) will be
      taken from a vein in one arm. Then the solution containing amino acids and nucleotides will
      be pumped slowly into your blood over the next 9-hour period. You will not be allowed to eat
      normal foods during this 9-hour period. However, you will be given a liquid nutritional
      formula to drink each hour during the day (starting after the first blood sample is taken).
      Approximately 14 blood samples (9 mL, less than 2 teaspoons each) and 8 breath samples will
      be taken from you while you receive the amino acid solution. In addition, at 6 times during
      the day you will be asked to lie quietly for 10 minutes while breathing into a special, clear
      hood that will measure your metabolic rate (the rate at which your body uses energy). You
      will remain at the GCRC for the entire 9-hour period, although you will be able to do limited
      activities (TV and reading). At the end of the 9-hour period, you will be able to go home and
      resume your activities. The following day you will resume eating meals prepared by the GCRC.
      While at the GCRC on days 3 and 4 after this procedure, you will be asked to give two more
      blood samples, one in the morning and one in the evening, and on day 5 you will be asked to
      give one blood sample in the morning (2 teaspoons each).

      The day after you are discharged from the GCRC, you will be asked to eat, for a period of
      approximately 4 weeks, a diet containing reduced amounts of vitamin B6 and limit the amount
      of caffeine (in coffee, tea, and carbonated drinks) you drink to one to two per day. Twice
      each day during this approximately 4 week period you will come to the General Clinical
      Research Center (GCRC) at Shands Hospital where you will eat your breakfast and evening
      meals. Also, you will be weighed once daily and have your heart rate, respirations and blood
      pressure monitored twice weekly. You will be given a sack lunch to be consumed wherever you
      choose. To be sure that you are getting adequate amounts of all nutrients except the vitamin
      being studied, you will be given a pill containing a vitamin and mineral supplement each day.
      Once each week (when you come into the GCRC to eat breakfast), you will have two teaspoons of
      blood taken from a vein in your arm to allow monitoring of your vitamin B6 status.

      After finishing approximately 4 weeks on this diet, you will participate again in the
      procedure to test how effectively vitamin B6 functions in helping your body use certain amino
      acids. Again, this will involve giving you a solution of certain amino acids by a vein in
      your arm over the course of 9 hours. As before, you will report to the GCRC at approximately
      8:00 p.m. on the evening before the procedure. You will spend the night at the GCRC and will
      not be allowed to eat after 9:00 p.m. The exact same procedures will be carried out as in the
      first admission.

      A few days prior to the second admission, while you are on the GCRC receiving your diet, you
      will have the same general health blood and urine tests collected as you had done before the
      first admission.

      When the amino acid solution has been completely given, approximately 9 hours, you will be
      able to go home and resume your activities. You will be asked to return to the GCRC to give
      blood samples on days 3, 4, and 5 after this procedure just as you did after the first
      admission. You will then be finished with the study. You will be given a 30 day supply of
      vitamin and mineral supplements to be taken daily to be sure that your body gets adequate
      amounts of vitamin B6 and other nutrients.

      After the laboratory analysis of blood samples from this study has been completed, if you
      agree, some of the samples will be kept in frozen storage for possible future analysis of
      nutrients or genetic information related to how your body processes nutrients. Please refer
      to the Addendum for Blood Storage and Future Testing attached.
    

Intervention name(s): Arm 1 glycine and leucine; Arm 2 Intervention of Serine and methionine infusion
Intervention type(s): Dietary Supplement; Dietary Supplement
Intervention description(s): Arm 1 glycine and leucine: A subset of 14 participants out of the total enrolled participants with an adequate nutritional status will undergo the tracer infusion of glycine and leucine amino acids protocol while an adequate vitamin pyridoxine, B6 status. After the first infusion each participant will then begin a diet low in vitamin B6 (<0.5 mg/d) for 4 weeks to achieve marginal B6 status followed by a repeat of fasting blood sampling and tracer infusion protocol.; Arm 2 Intervention of Serine and methionine infusion: A subset of 14 participants out of the total enrolled participants. Received an adequate nutritional status will undergo the tracer infusion of methionine and serine amino acids protocol while in adequate vitamin pyridoxine, B6 status. After the first infusion each subject will then begin a diet low in vitamin B6 (<0.5 mg/d) for 4 weeks to achieve marginal B6 status followed by repeat of fasting blood sampling and tracer infusion protocol.
Other intervention name(s): amino acids; B6; leucine; methionine; deficieny; B6 deficiency; methionine; serine
Intervention MeSH terms: Glycine
Allocation: Non-Randomized
Intervention model: Single Group Assignment
Masking: None (Open Label)
Primary purpose: Diagnostic
Keywords: Vitamin B6; Vitamin B6 deficiency; Amino acids; pyridoxine