Comparisons of uptake and cell surface binding among pyridoxal,pyridoxine, and pyridoxamine in RAW264.7 cells

ObjectiveVitamin B6 (B6) suppresses the expression of cyclooxygenase-2 stimulated by lipopolysaccharide in mouse macrophage RAW264.7 cells. The greatest effect is recognized for pyridoxal (PL) compared with pyridoxamine (PM), pyridoxine (PN), and pyridoxal 5′-phosphate (PLP). However, it has not been elucidated why PL has the strongest effect. We compared the uptakes and cell surface interactions among PL, PM, PN, and PLP in RAW264.7 cells.MethodsCyclo-oxygenase-2 mRNA expression was evaluated by real-time polymerase chain reaction. Intracellular B6 concentrations were measured by high-performance liquid chromatography. Interactions of B6s with the cell surface were analyzed using a surface plasmon resonance biosensor. B6 uptake speeds were measured using [³H]-PN.ResultsThe intracellular PLP levels did not change significantly when cells were cultured in medium containing PL, PM, PN, or PLP. Only PL interacted with the cell surface. Although PM and PN were associated with the cell surface, their binding was only recognized during sample loading. After the change to phosphate buffered saline after sample loading, the binding resonances of PM and PN returned to baseline, whereas that of PL did not. Uptake of [³H]-PN was inhibited by non-labeled PN, PL, or PLP, but not PM, at 1 μM. The inhibition rate of PL was higher than those of PN and PLP.ConclusionThe inhibition of cyclo-oxygenase-2 mRNA expression by PL may be related to the cell surface interaction of PL, rather than the intracellular PLP level. The uptake mechanism for PN and PL may differ from that for PM.     

The relationship between plasma and red cell concentrations of vitamins thiamine diphosphate, flavin adenine dinucleotide and pyridoxal 5-phosphate following elective knee arthroplasty

BACKGROUND & AIMS: Water soluble vitamins B1, B2 and B6 are essential precursors for a wide variety of coenzymes involved in intermediary metabolism and their status is usually assessed from blood samples. The aim of the study was to examine the relationship between plasma and intra-cellular B-vitamins following the systemic inflammatory response of surgery. METHODS: Patients (n = 10) who underwent an elective knee arthroplasty, had venous blood samples withdrawn pre-operatively and at 12, 24, 48, 72 and 168 h after the start of surgery for the analysis of circulating concentrations of C-reactive protein and albumin and also plasma and/ or red cell thiamine diphosphate (TDP), flavin adenine dinucleotide (FAD), pyridoxal 5-phosphate (PLP) as indicators of vitamins B1, B2, and B6 status respectively. RESULTS: Pre-operative, baseline vitamin assessments were all within population reference ranges. Over the study period of 0-168 h there was a significant increase in circulating C-reactive protein concentrations (peak 48 h, P < 0.001) and a significant fall in albumin concentrations (trough 48 h, P < 0.001). Plasma FAD and PLP concentrations fell transiently (P < 0.001) by approximately 40% reaching their nadir at approximately 48 h. CONCLUSIONS: The results of the present study indicate that plasma concentrations of FAD and PLP are transiently reduced following an inflammatory insult and therefore unlikely to be a reliable measure of status in the presence of a systemic inflammatory response. It may be that during such a response red cell concentrations provide a more reliable measure.     

Plasma pyridoxal 5'-phosphate is a significant indicator of immune responses in the mechanically ventilated critically ill

OBJECTIVES: This study assessed the effect of vitamin B6 status on immune responses in mechanically ventilated, critically ill patients and compared the results with those of healthy controls. METHODS: This was designed as a cross-sectional observational study. Forty patients in the intensive care unit successfully completed this study. Vitamin B6 intake was recorded for 8 d. Severity of illness (Second Acute Physiology and Chronic Health Evaluation score) was recorded. Thirty-eighty healthy controls were recruited from the physical check unit of Taichung Veterans General Hospital (Taichung, Taiwan). All control subjects were given instruction on how to complete a 24-d diet recall. Vitamin B6 status was assessed by direct measures (plasma pyridoxal 5'-phosphate [PLP] and 4-pyridoxic acid) and indirect measures (erythrocyte alanine and aspartate aminotransferase activity coefficients). Levels of serum albumin, hemoglobin, hematocrit, high-sensitivity C-reactive protein, and immune responses (white blood cell, neutrophil, total lymphocytes, T lymphocytes [CD3], B lymphocytes [CD19], T-helper cells [CD4], and suppressor cells [CD8]) were determined. RESULTS: Critically ill patients had sufficient vitamin B6 intake but showed marginal PLP deficiency (20.9 +/- 1.5 nmol/L). In addition, critically ill patients had significantly lower and abnormal immune responses than did healthy controls. There was no significant correlation of vitamin B6 intake and erythrocyte alanine and aspartate aminotransaminase activity coefficients with immune indices. Plasma PLP concentration was strongly negatively correlated with high-sensitivity C-reactive protein level. However, plasma PLP was significantly associated with immune responses after adjustment for age, sex, high-sensitivity C-reactive protein, and the other four vitamin B6 indicators. CONCLUSIONS: Plasma PLP is a significant indicator of immune responses in human subjects. Further research is warranted to study whether vitamin B6 supplementation in critically ill patients improves their immune responses.     

Vitamin B6 intakes and status of mechanically ventilated critically ill patients in Taiwan

OBJECTIVE: To assess vitamin B6 intake and status of critically ill patients. The relationship between vitamin B6 status indicators and the severity of illness and outcome in these patients was also examined. DESIGN: Prospective clinical study. SETTING: The study was performed at the Taichung Veteran General Hospital, in the central part of Taiwan. SUBJECTS: Ninety-four patients in the intensive care unit (ICU) entered the study and 46 patients successfully completed this study. INTERVENTIONS: No intervention. MAIN OUTCOME MEASURES: Vitamin B6 intake was recorded for 14 days. Vitamin B6 status was assessed by direct measures (plasma pyridoxal 5'-phosphate (PLP), pyridoxal (PL), and urinary 4-pyridoxic acid (4-PA)) and indirect measures (erythrocyte alanine (EALT-AC) and aspartate (EAST-AC) aminotransaminase activity coefficient). The severity of illness (APACHE II score), the length of ventilation dependency, and the length of ICU and hospital stay were recorded. RESULTS: Patients had an adequate mean vitamin B6 intake (16.26+/-19.39 mg) during the 14 day study. Mean vitamin B6 intake was significantly higher on day 14 than on day 1 (P<0.001). However, plasma PLP and PL concentrations significantly decreased at the 14th day after admission (P<0.05). Erythrocyte alanine aminotransaminase activity coefficient and EAST-AC did not change significantly. Urinary 4-PA significantly increased at the 14th day (P<0.001). No significant relationships were found between APACHE II scores and clinical outcomes (the length of ICU and hospital stay, the length of ventilation dependency) of patients, vitamin B6 intake or status indicators. CONCLUSIONS: Critically ill patients received nutritional support in the ICU, and had sufficient mean vitamin B6 intake and adequate vitamin B6 status. Therefore, the severity of illness and the results should not be affected by vitamin B6 status. However, we have noted that plasma PLP and PL concentrations significantly decreased while vitamin B6 intake significantly increased on day 14. Critical clinical conditions and complex metabolism in the critically ill may account for the reduction of plasma PLP and PL. Since vitamin B6 deficiency causes profound effects on immune system function, dietary or supplemented vitamin B6 intake is suggested for hospitalized patients.     

Vitamin B6 supplementation increases immune responses in critically ill patients

OBJECTIVE: To investigate whether vitamin B6 supplementation has a beneficial effect on immune responses in critically ill patients. DESIGN: A single-blind intervention study. SETTING: The study was performed at the Taichung Veterans General Hospital, the central part of Taiwan. SUBJECTS: Fifty-one subjects who stayed over 14 days in the intensive care unit completed the study. Subjects were not treated with any vitamin supplement before the intervention. INTERVENTIONS: Patients were randomly assigned to one of three groups, control (n = 20), a daily injection of 50 mg vitamin B-6 (B6 -50, n=15), or 100 mg vitamin B-6 (B6 -100, n = 16) for 14 days. MAIN OUTCOME MEASURES: Plasma pyridoxal 5'-phosphate (PLP), pyridoxal (PL), 4-pyridoxic acid (4-PA), erythrocyte alanine (EALT-AC) and aspartate (EAST-AC) aminotransaminase activity coefficient, and urinary 4-PA were measured. The levels of serum albumin, hemoglobin, hematocrit, high-sensitivity C-reactive protein (hs-CRP) and immune responses (white blood cell, neutrophils, total lymphocytes count (TLC), T- (CD3) and B-(CD19) lymphocytes, T-helper (CD4) and suppressor (CD8) cells) were determined. RESULTS: Plasma PLP, PL, 4-PA and urinary 4-PA concentrations significantly increased in two treated groups. T-lymphocyte and T-helper cell numbers and the percentage of T-suppressor cell significantly increased on day 14 in the B6 -50 group. Total lymphocyte count, T-helper and T-suppressor cell numbers, the percentage of T-lymphocyte cells and T-suppressors significantly increased in the B6 -100 group at the 14th day. There were no significant changes with respect to immune responses in the control group over 14 days. CONCLUSIONS: A large dose of vitamin B6 supplementation (50 or 100 mg/day) could compensate for the lack of responsiveness of plasma PLP to vitamin B6 intake, and further increase immune response of critically ill patients. SPONSORSHIP: This study was supported by the National Science Council, Taiwan, Republic of China (NSC-92-2320-B-040-026).     

Neuroprotective effects of pyridoxal phosphate and pyridoxal against ischemia in monkeys

Previously, in monkeys undergoing 20 min whole brain ischemia we demonstrated that the activated calpain-induced lysosomal disruption with the resultant leakage of cathepsins B and L, causes neuronal death in the cornu Ammonis (CA) 1 sector on day 5. Selective cathepsin inhibitors significantly protected ischemic CA1 neurons from delayed necrosis. Recently, pyridoxal phosphate (PLP) and pyridoxal (hydrochloride) (PL) were demonstrated to inhibit cathepsins B and L in vitro, because the active aldehyde at position 4 of the pyridine ring has an affinity for the active site -SH of cysteine residues of cathepsins. Here, we studied whether PLP and PL can, in vivo, protect monkey CA1 neurons from ischemic insult. In monkeys undergoing 20 min whole brain ischemia, 15 mg/kg body weight/day of drugs were intravenously injected for 10 days before and after the ischemic insult. Histological analysis of the surviving CA1 neurons was done using the hippocampus resected on day 5 after ischemia. For PLP or PL, approximately 17% (P = 0.0639) or 54% (P < 0.0001) of the total population (100%) of control CA1 neurons were, respectively, saved from the ischemia-induced neuronal death, showing a remarkable contrast to the surviving neurons (approximately 3.9%) in non-treated monkeys. These data suggested that PL (perhaps PLP intracellularly) is useful as a novel neuroprotectant in primates.     

The relationship between plasma and red cell B-vitamin concentrations in critically-ill patients

BACKGROUND AND AIMS: Low vitamin B-complex status has been associated with poorer outcome in critically-ill patients. However, these findings have been based on indirect methods. Using direct methods for assessing vitamin status, we examined the effect of B-complex vitamin supplementation by measuring plasma and red blood cell B1, B2 and B6-vitamin concentrations in critically-ill patients. METHODS: Thiamine diphosphate (TDP), flavin adenine dinucleotide (FAD) and pyridoxal phosphate (PLP) concentrations were measured in plasma and red cells of normal subjects (n=49) and ITU patients (n=41). RESULTS: Compared with the normal subjects, critically-ill patients had higher C-reactive protein and lower albumin concentrations (P<0.001). Also, plasma FAD and PLP were lower (P<0.001) and red cell concentrations of both were higher (P<0.01) in critically-ill patients. Critically-ill patients were grouped according to whether (n=23) or not (n=18) they had been supplemented with B-complex vitamins. Compared with non-supplemented group, the supplemented group had significantly higher red cell TDP and PLP concentrations (P<0.01). Plasma FAD and PLP concentrations did not differ significantly between the groups. CONCLUSIONS: The results of the present study suggest that direct measurements of red cell FAD and PLP are more responsive to supplementation than plasma measurements in the critically-ill patient.     

Plasma pyridoxal and pyridoxal 5'-phosphate concentrations in response to ingestion of water or glucose polymer during a 2-h run

The effects of ingesting a glucose polymer solution (GP) or water (W) on plasma pyridoxal 5'-phosphate (PLP) and pyridoxal (PL) concentrations were compared in six men (age: 30 +/- 2 y; VO2max: 57.4 +/- 3.2 mL.kg-1.min-1) under running (R) and control (C) conditions. Subjects ran for 2 h at 60-65% of VO2max for R and remained standing for C. For both R and C, 200 mL W or GP was ingested before (0-time) and every 30 min while running (30, 60, and 90 min). Plasma PLP decreased to 95% and 87% of 0-time at 180 min for WC and GPC and increased to 126% and 119% at 90 min and to 124% and 119% at 120 min for WR and GPR. By 60 min postrun, plasma PLP was 98% (WR) and 101% (GPR) of 0-time. There were no significant differences between W and GP conditions. Changes in PLP were not related to plasma volume or blood glucose, free fatty acids, lactate, alkaline phosphatase, aspartate aminotransferase, or alanine aminotransferase. No significant changes in plasma PL were noted. Exercise induces an increase in plasma PLP, perhaps due to transfer of B-6 vitamers from liver to skeletal muscle.     

Intestinal absorption of pyridoxal 5'-phosphate at physiological levels in rats

The intestinal absorption of pyridoxal 5'-phosphate (PLP) at physiological levels (10(-7) -10(-6) M) was studied in comparison with that of pyridoxal (PL) in rat, using in vitro everted sac and an intestinal preparation that permitted continuous in situ collection of mesenteric venous blood. After PLP administration (10(-6) -10(-3) M) in situ, larger amounts of PLP were found in the mesenteric venous plasma than after PL administration at the same dose. The amount of PLP found in the mesenteric venous plasma was dependent on its dose at lower concentrations up to 10(-4) M but became independent at higher concentrations. After PL administration at various doses, the amount of PL found in the mesenteric venous blood increased linearly with the dose. When various concentrations of PLP were added to the mucosal side, under the in vitro condition with protection from alkaline phosphate hydrolysis, PLP was detected in the serosal side and the extent of PLP transport was dependent on the initial concentration of PLP in the mucosal side. When various concentrations of PL were added to the mucosal side, the extent of PL transport was independent of the initial concentration of PL in the mucosal side. In rat pretreated with actinomycin D, PLP transport in vitro was inhibited but not that of PL. N2-induced anoxia and pyridoxamine 5'-phosphate and anion transport inhibitor (4,4'-diisothiocyanostilben-2,2'-disulfonic acid disodium salt) showed no effect on PLP transport. These results suggest that PLP can be absorbed in the phosphorylated form and imply the presence of a saturable process for direct absorption of PLP itself and a diffusive process for PL absorption. In addition, the result of the in vivo neonatal experiment suggests that the neonatal intestine also can transport PLP in phosphorylated form.     

The relationships between plasma and red cell vitamin B2 and B6 concentrations and the systemic and local inflammatory responses in patients with colorectal cancer

B vitamins have been implicated in cancer pathogenesis. It is therefore of interest that plasma B6 falls as part of the systemic inflammatory response (SIR), whereas red cell concentrations do not. The modified Glasgow Prognostic Score (mGPS) is a validated inflammation-based prognostic score that consists of a combination of albumin and C-reactive protein concentrations. The aim of this study was to examine the relationships between the concentrations of plasma and red cell vitamin B concentrations, the local and systemic inflammatory response in patients with colorectal cancer. Preoperative venous blood of 108 patients with colorectal cancer were analyzed for C-reactive protein, albumin, flavin adenine dinucleotide (FAD), and pyridoxal phosphate (PLP), and lymphocyte counts. Pathological slides were retrieved for assessment of inflammatory cell infiltration. Increasing mGPS was associated with lower plasma PLP concentrations (P < 0.01) but not plasma and red cell FAD and red cell PLP concentrations. Increasing tumor stage was associated with the presence of venous invasion (P < 0.01) and low-grade inflammatory cell infiltrate (P < 0.05) but not the SIR, FAD, or PLP concentrations. A low-grade inflammatory cell infiltrate was not significantly associated with any other parameter. The presence of a SIR was associated with lower concentrations of plasma PLP but not red cell PLP concentrations in patients with colorectal cancer. Neither FAD and PLP were associated with the tumor inflammatory cell infiltrate.     

Adequacy of maternal pyridoxine supplementation during pregnancy in relation to the vitamin B6 status and growth of neonates at birth

To evaluate the adequacy of maternal pyridoxine supplementation during pregnancy for both maternal and neonatal status at birth, vitamin B6 status, assessed by plasma pyridoxal phosphate (PLP), pyridoxal (PL) and total aldehyde vitamer (PLP + PL) concentrations, and the growth of neonates, including weight, length, head and chest circumferences, were examined for 209 neonates whose mothers were supplemented with 0, 1, 2 or 3 mg pyridoxine.HCl (PN.HCl)/d during pregnancy. Maternal PN.HCl supplementations were positively correlated to both maternal (r = 0.62) and cord (r = 0.78) plasma PLP concentrations (p < 0.0001). Mothers supplemented with 2 or 3 mg/d PN.HCl had significantly higher plasma concentrations of PLP and total B6 aldehyde vitamer in maternal and cord blood compared with those receiving 0 or 1 mg PN.HCl/d. A growth benefit for neonates whose mothers had maternal and cord plasma PLP concentrations > or = 40 nM was revealed by the maternal supplementation of 2 mg PN.HCl/d during pregnancy. Thus, in healthy pregnant women, according to our study, a daily supplement of 2 mg PN.HCl provides the adequacy of maternal and neonatal vitamin B6 status and the satisfactory growth of neonates at birth.     

B-6 vitamers and 4-pyridoxic acid in the plasma, erythrocytes, and urine of postmenopausal women

BACKGROUND: Although many studies have reported reduced vitamin B-6 status with aging, little information is available about the specific effects of menopause. OBJECTIVE: We aimed to examine vitamin B-6 metabolism in premenopausal and early postmenopausal women. DESIGN: We examined dietary intake and vitamin B-6 metabolites in the plasma, erythrocytes, and urine of 30 premenopausal women (x +/- SD age: 41.9 +/- 4.8 y) and 30 women (aged 54.0 +/- 3.8 y) who were 4.0 +/- 1.4 y past menopause. RESULTS: Vitamin B-6 intake in the postmenopausal group (1.97 +/- 0.40 mg/d) was significantly greater than that in the premenopausal group (1.63 +/- 0.50 mg/d). Plasma pyridoxal phosphate (PLP) and pyridoxal concentrations and erythrocyte PLP, pyridoxal, and pyridoxamine phosphate concentrations were in the normal range in both groups and did not differ significantly between the 2 groups. Plasma and erythrocyte 4-pyridoxic acid (4-PA) concentrations were significantly higher in the postmenopausal group than in the premenopausal group, which may have been due at least partly to the slightly higher vitamin B-6 intake of the former group. Erythrocyte 4-PA was correlated (r = -0.37, P < 0.01) with serum estradiol in both groups. Urinary 4-PA did not differ significantly between the 2 groups. The serum phosphate concentration was higher in the postmenopausal group than in the premenopausal group, and it was correlated (r = 0.40, P < 0.01) with plasma PLP. Inhibition of alkaline phosphatase by the increased phosphate may help to increase plasma PLP. CONCLUSION: Menopause may not necessarily be associated with a decrease in vitamin B-6 status.     

Pyridoxine supplementation of lactating mothers: relation to maternal nutrition status and vitamin B-6 concentrations in milk

Vitamin B-6 status, assessed by plasma pyridoxal phosphate (PLP) concentrations, and vitamin B-6 concentrations in breast milk were examined in 47 lactating mothers supplemented with different amounts of pyridoxine.HCl (PN.HCl) during pregnancy and the first 6 mo of lactation. PLP concentrations in cord blood and maternal plasma at 2 d postpartum and vitamin B-6 concentration in colostrum were positively correlated with the amount of PN.HCl supplementation prenatally (r = 0.71, p less than 0.001; r = 0.74, p less than 0.001; and r = 0.78, p less than 0.001, respectively). Correlations between the amounts of PN.HCl supplementation postnatally and plasma PLP concentrations increased with the length of supplementation. Plasma PLP concentrations were also correlated with vitamin B-6 concentrations of milk samples, which were obtained on the same day as plasma. PN.HCl supplements between 2.5 and 4.0 mg/d (2.1-3.4 mg PN equivalents) ensured vitamin B-6 adequacy of the mother and maintained relatively saturated concentrations of vitamin B-6 in breast milk.     

Relationship between blood, liver and brain pyridoxal phosphate and pyridoxamine phosphate concentrations in mice

Plasma pyridoxal 5'-phosphate (PLP) concentrations are considered to be the most reliable single indicator of vitamin B-6 nutritional status and are thought to reflect tissue PLP and pyridoxamine 5'-phosphate (PMP) levels. We investigated the relationship between dietary level of pyridoxine hydrochloride (PN-HCl) and concentrations of PLP in blood and PLP and PMP in liver and brain of mice. Female heterogeneous stock mice, 60 to 90 d old, were fed purified diets containing 0.5, 1.0, 2.0, 3.0, 5.0, or 7.0 mg PN-HCl/kg diet for 5 wk. PLP and PMP concentrations were determined by a spectrophotometric apotryptophanase assay. PLP content of plasma, erythrocytes, whole blood, liver and brain and PMP levels in liver and brain were highly correlated with dietary level of PN-HCl (r values ranged from 0.81 to 0.94, n per correlation = 32 to 43). By using the entire range of dietary levels of PN-HCl, both plasma and erythrocyte PLP were found to be significantly correlated with tissue PLP and PMP concentrations. For any one dietary level, however, correlations between plasma or erythrocyte PLP and tissue PLP and PMP concentrations were low and nonsignificant. These results suggest that plasma PLP levels may be suitable to determine vitamin B-6 status of populations, but not to reliably predict tissue concentrations of PLP or PMP in individuals.     

A comparison of plasma pyridoxal 5'-phosphate concentrations in capillary (finger prick) and venous blood

The purpose of our study was to investigate the validity of using capillary blood plasma to estimate the pyridoxal 5'-phosphate (PLP) concentration of venous blood plasma. To accommodate the small volumes of capillary blood usually obtained, we modified the experimental conditions of the L-tyrosine apodecarboxylase (TDC) assay for PLP by increasing both the specific activity of 14C-tyrosine and the reaction incubation time. Plasma PLP concentrations determined by the TDC assay and the micro-modified TDC assay were highly correlated (r = 0.995, p = 4.0 X 10(-6)). Using the micro-modified TDC assay, we observed no significant difference between the plasma PLP concentrations of venous and capillary (finger prick) blood from 10 healthy adults. Thus, capillary blood plasma can be used to estimate the PLP concentration of venous blood plasma. This observation will aid in verifying data concerning the vitamin B-6 status assessment of individuals as determined by capillary blood plasma PLP concentrations.     

Depressed plasma pyridoxal phosphate concentrations in adult asthmatics

In 15 adult patients with bronchial asthma, plasma and erythrocyte pyridoxal phosphate (PLP) concentrations were significantly lower than in 16 controls (P less than 0.0001 and P less than 0.005, respectively). Oral supplementation of seven asthmatics with 50 mg pyridoxine as pyridoxine X HC1 twice daily failed to produce a sustained elevation of PLP in either the plasma or erythrocytes. However, all subjects reported a dramatic decrease in frequency and severity of wheezing or asthmatic attacks while taking the supplement. The reasons for the failure of a uniform elevation in plasma and erythrocyte PLP concentration and for the apparent beneficial effects of pyridoxine supplementation on the asthmatic symptoms of the patients are unknown at present.     

Cheek cell phospholipids in human infants: a marker of docosahexaenoic and arachidonic acids in the diet, plasma, and red blood cells

BACKGROUND: Assessment of essential fatty acid status requires collection of blood or adipose tissue samples. However, these invasive techniques cannot always be used in studies involving infants, young children, or subjects from whom it is difficult to obtain blood. A body tissue that is easy to access is the buccal mucosa (cheek cells). OBJECTIVE: The objective was to investigate the degree to which fatty acids of cheek cells reflect the fatty acid content of plasma, red blood cells, and the diet. DESIGN: Thirty-one infants aged 12 mo were enrolled. Five infants were fed human milk and 26 infants received formulas that provided a wide range of arachidonic acid and docosahexaenoic acid (DHA) intakes. Cheek cells were collected on a small piece of gauze by gently swabbing the inside of the cheek 3 times. Lipids were extracted from the gauze and the phospholipid fatty acid content of the cheek cells was determined. RESULTS: Cheek cell DHA and arachidonic acid in phospholipids were significantly correlated with DHA and arachidonic acid in plasma [r = 0.61 (P < 0.001) and r = 0.37 (P <0.05), respectively], red blood cells [r = 0.58 (P < 0.001) and r = 0.37 (P < 0.05), respectively], and the diet [r = 0.65 (P < 0.001) and r = 0. 51 (P < 0.01), respectively]. CONCLUSIONS: Given these correlations and the ease and noninvasive nature of this technique, cheek cell fatty acids may serve as a marker of the essential fatty acid content, especially of DHA and arachidonic acid, in plasma, red blood cells, and the diet.     

The Relationships Between Plasma and Red Cell Vitamin B2 and B6 Concentrations and the Systemic and Local Inflammatory Responses in Patients With Colorectal Cancer

B vitamins have been implicated in cancer pathogenesis. It is therefore of interest that plasma B6 falls as part of the systemic inflammatory response (SIR), whereas red cell concentrations do not. The modified Glasgow Prognostic Score (mGPS) is a validated inflammation-based prognostic score that consists of a combination of albumin and C-reactive protein concentrations. The aim of this study was to examine the relationships between the concentrations of plasma and red cell vitamin B concentrations, the local and systemic inflammatory response in patients with colorectal cancer. Preoperative venous blood of 108 patients with colorectal cancer were analyzed for C-reactive protein, albumin, flavin adenine dinucleotide (FAD), and pyridoxal phosphate (PLP), and lymphocyte counts. Pathological slides were retrieved for assessment of inflammatory cell infiltration. Increasing mGPS was associated with lower plasma PLP concentrations (P < 0.01) but not plasma and red cell FAD and red cell PLP concentrations. Increasing tumor stage was associated with the presence of venous invasion (P < 0.01) and low-grade inflammatory cell infiltrate (P < 0.05) but not the SIR, FAD, or PLP concentrations. A low-grade inflammatory cell infiltrate was not significantly associated with any other parameter. The presence of a SIR was associated with lower concentrations of plasma PLP but not red cell PLP concentrations in patients with colorectal cancer. Neither FAD and PLP were associated with the tumor inflammatory cell infiltrate.     

A new perspective in the assessment of vitamin B-6 nutritional status during pregnancy in humans

Plasma pyridoxal-5'-phosphate (PLP) concentration has been suggested as a valid indicator to assess vitamin B-6 nutritional status. Animal and human studies have shown that plasma PLP concentrations decrease progressively during pregnancy and large doses of vitamin B-6 supplementation are required to maintain plasma PLP at early or prepregnant levels. PLP is known to be hydrolyzed to pyridoxal (PL) by alkaline phosphatase (ALP), resulting in an inverse relationship between PLP and ALP. The object of this study was to compare the PLP-PL equilibrium in a group of healthy pregnant females with that of an age-matched nonpregnant control group from a similar socioeconomic background. The mean plasma PLP level was 37% lower, (P less than 0.0001), whereas the mean PL level was almost 90% higher (P less than 0.001) in the pregnant group than in the nonpregnant control group. The total amount of plasma PLP and PL levels, however, did not differ significantly (P greater than 0.24) between the two groups. Because the PL vitamer is regarded as the ultimate transport form of vitamin B-6, it may serve as a readily available source of vitamin B-6 to meet possible increased metabolic demands. Therefore, the estimation of plasma PLP alone does not permit an accurate assessment or understanding of the nutritional status and the physiology of vitamin B-6 in conditions associated with altered vitamin B-6 homeostases.     

Occupancy of glycoprotein IIb/IIIa by B-6 vitamers inhibits human platelet aggregation

Vitamin B-6 inhibits platelet aggregation. However, the effect of the occupancy of GPIIb/IIIa, a major receptor responsible for aggregation on platelet membranes, by B-6 vitamers on platelet aggregation is unknown. This study was carried out to quantify GPIIb/IIIa occupancy in platelets treated with B-6 vitamers [pyridoxal-5-phosphate (PLP); pyridoxal (PL); pyridoxine (PN); pyridoxamine (PM)], using a monoclonal antibody-based assay, by flow cytometry. Antibody binding was compared with inhibition of platelet aggregation. PLP, PL, PN and PM occupied GPIIb/IIIa with dissociation constants of 1.83 +/- 1.15, 19.43 +/- 7.86, 3.63 +/- 1.67 and 10.89 +/- 2.93 mmol/L, respectively. Occupancy of GPIIb/IIIa by the four B-6 vitamers was negatively correlated with platelet aggregation (r = -0.90 to -0.94, P < 0.001). The concentrations of the four B-6 vitamers that inhibited maximal platelet aggregation were in the order of PLP < PN or =80% of the GPII/IIIa receptor. Platelet aggregation was inhibited by B-6 vitamers via the occupancy of GPIIb/IIIa with the potency of PLP > PN > PM > PL.