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Effect of sodium ascorbate concentration on the stability of samples for determination of serum folate levels.

Sodium ascorbate can be used as a preservative of patient samples for folate assay when freezing of serum is impractical. To evaluate the effect of sodium ascorbate on folate levels in human serum, it was added to pooled human sera in 1 g/l increments from 0 to 10 g/l serum. Free folate levels remained constant when the sodium ascorbate concentration was 6 g or less per liter of serum. At more than 6 g/l serum, free folate levels decreased. Bound folate levels increased when sodium ascorbate levels were 4 g/l or less, but remained stable at more than 4 g/l. A sodium ascorbate concentration of 5.0 +/- 1 g/l serum provided optimal preservation of folate in patient samples, indicated by obtaining constant values for four days when serum was kept at room temperature.

Ascorbic Acid

Serum folate levels in horses, with particular reference to the English thoroughbred.

The assay of serum folate has been used to investigate the folate status of thoroughbreds in training to race, in-foal thoroughbred mares at stud and crossbred ponies kept permanently at grass. Throughbreds in training, with no obvious haematological or clinical abnormalities, had lower serum folate levels (mean 3.3, range 1.5 to 6.1 microgram per litre) compared with thoroughbred mares at stud (mean 10.6, range 6.4 to 15.8 microgram per litre) and crossbred ponies at grass (mean 10.9, range 7.4 to 16.6 microgram per litre. Intensive exercise, over a period of six months, significantly decreased serum folate levels (P less than 0.001) and significantly increased the levels of bilirubin (P less than 0.0001) and PCV (P less than 0.0002) in a group of 30 thoroughbred racehorses. Haptoglobin levels showed no significant changes.

Animal Feed

Serum folate levels. Comparison of microbiologic assay and radioisotope kit methods.

Serum folate levels were measured prospectively in 72 patients and six normal controls using the Lactobacillus casei microbiologic method and a commercially available sequential-binding radioassay kit method. Values between 0 and 14 ng/ml (0 and 31.78 nmol/l) (clinically significant range) obtained by the microbioassay and sequential-binding radioassay were analyzed statistically. The data suggest that comparable values are obtained by the two technics (Y = 0.9 X where Y = radioassay value). Data obtained by the sequential-binding radioassay and a competitive protein-binding radioassay kit were then compared and subjected to statistical analysis. Values obtained by the two radioassay methods were found to be comparable clinically, but significantly different statistically. The authors conclude that these commercially available radioassay kits for measuring serum folate yield reliable values that can be applied to the clinical evaluation of patients with anemia, and can be readily performed in a clinical radioisotope laboratory.

Biological Assay

Haematological profile in leprosy. Part II--Relationship to severity of disease and treament status.

321 adult male lepromatous leprosy patients were studied for relationship between haematological findings, severity of disease and duration of treatment. Significant changes were noticed in relation in haemoglobin concentration, serum vitamin B12 and serum folate levels, serum albumin and globulin. No significant changes were observed in serum iron levels in relation to disease and treatment status. With rising bacterial load, there was a trend towards lower haemoglobin concentration, higher vitamin B12 level and lowered serum folate levels. Serum albumin showed a significant decline, while serum globulin showed a significant rise. The findings are discussed in relation to replacement of bone marrow by lepromatous tissue as well as possible interference in the metabolism of haematinics by M. leprae. The exact mechanism of neurlogical deficit in leprosy in relation to deficiency of vitamin B12 and folic acid need to be further elucidated.

Folic Acid

Serum erythrocyte folate levels in thalassaemic patients in Thailand.

The mean serum and adjusted red cell folate levels, as measured by microassay using Lactobacillus casei, respectively were 8.87 +/- SD 3.28 microgram/l and 436 +/- SD 107 microgram/l in 76 normal subjects, 4.22 +/- SD 2.70 microgram/l and 182 +/- SD 114 microgram/l in 55 patients with beta-thalassaemia/Hb E disease and 6.36 +/- SD 2.95 microgram/l and 320 +/- 158 microgram/l in 37 patients with Hb H disease. The mean serum and adjusted red cell folate values of the patients with beta-thalassaemia/Hb E disease and the mean serum folate value of the patients with Hb H disease were significantly lower than those of the normal subjects (P less than 0.001). 33% of the beta-thalassaemia/Hb E patients and 8% of the Hb H patients showed low serum folate levels (less than 3 microgram/l) whereas 84% of the former and 45% of the latter showed low adjusted red cell folate levels (less than 270 microgram/l). The group of beta-thalassaemia/Hb E disease with low serum folate levels had lower mean haemoglobin concentration and lower mean adjusted red cell folate level than the group with normal serum folate levels. Since the Thai diets have high folate content, the observed low serum and erythrocyte folate levels in thalassaemic patients most likely occur from massively increased erythropoiesis. Folate, 5 mg/d, is now routinely prescribed to such patients especially to those with severe anaemia.

Adolescent

Folic acid binding protein and folate balance in uremia.

To determine the incidence and importance of folate deficiency in uremia, we studied 41 patients who had chronic uremia but who were not receiving hemodialysis. Serum folate level was assayed by microbiological, whole serum radioassay, and heat-extracted radioassay techniques. Mean serum folate level, as determined with the Lactobacillus casei method, was 6.9 ng/ml (normal, more than 3.0 ng/ml). The mean heat-extracted radioassay serum folate level was 6.6 ng/ml. Only 10% of our patients had subnormal serum folate values, as determined with these techniques. No cases of megaloblastic anemia were discovered, and the hematologic profiles correlated with L casei and heat-extracted radioassay serum folate values. The mean serum folic acid binding protein (FABP) level was significantly greater for the uremic patients than for control patients (26% vs 9.9%, respectively; P = .0005). Our results show that elevated levels of serum FABP spuriously depress the serum folate level, as determined with the whole serum radioassay technique, but apparently do not retard delivery of folate to the tissue in vivo.

Creatinine

Serum vitamin B 12 and folic acid levels in patients with fasciolopsiasis.

Serum vitamin B12, serum and red cell folate concentrations and vitamin B12 absorption were studied in 100 patients with fasciolopsiasis. A mean value of serum vitamin B12 level in the patient group was found to be significantly lower than that of normal subjects and 14% of these patients had serum vitamin B12 level less than 100 pg/ml. Serum UBBC and TBBC levels in the patients were significantly higher than those of the normal subjects. Serum TCI and TCIII increased significantly while TCII decreased. Vitamin B12 absorption was found to be impaired in 3 of 9 patients studied. There was no relationship seen between serum vitamin B12 level and vitamin B12 absorption. The mean values of serum folate and red cell folate levels in the patient group were significantly lower than those of normal subjects. Fifteen of 100 patients (15%) had serum folate level less than 3 ng/ml, while all of them had red cell folate higher than 100 ng/ml. Serum folic acid binding protein levels (FABP) in these patients, were not significantly different from those of normal subjects.

Child

Folate levels among various populations in central Iran.

Serum folic acid levels (Lactobacillus casei assay) were determined in 467 subjects representing various socioeconomic classes from rural and urban central Iran. Of the subjects 170 were pregnant females in different trimesters. The mean serum folate levels of the various groups studied ranged from 7.5 to 11.1 ng/ml and are within the normal reported range of 5.9 to 21.0 ng/ml. Only two subjects had serum folate levels below 3.0 to 5.9 ng/ml. In anemic subjects (hematocrit is less than 37%), red blood cell folate levels were also determined and were significantly higher than nonanemic controls (558 +/- 56 versus 346 +/- 35 ng/ml of packed red blood cells, P is less than 0.01). This difference may be related to underutilization of folate in patients with iron deficiency (a common metal deficiency in this area). A probable explanation for the well maintained serum folate levels in late pregnancy as well as in other populations studied in this report may be the high dietary intake of Iranian bread made from wheat flour of high extraction rate.

Adolescent

Geriatric mental function and serum folate: a review and survey.

A review of the growing body of information regarding folic acid biochemistry indicates a relationship between folate metabolism and mental disturbance. Clinical correlations between altered mental functions associated with aging and lowered serum folate values are found to suggest that further studies are warranted. The fasting serum folate levels of two groups of subjects 65 years of age and older are compared after excluding the influence of drugs, alcohol, and gastrointestinal disease. The results show that those elderly persons with good dietary habits and mental acuity had no lowered serum folate levels. Thus, the suggestion that serum folate level normally falls with age is not confirmed. Additionally, it was found that 35% of similarly aged subjects with combined mental dysfunction and poor dietary habits had lowered serum folate values.

Aged

Bird-fancier's lung and jejunal villous atrophy.

Sixteen patients with bird-fancier's lung were screened for evidence of coeliac disease by assessing their clinical features, red-bloodcell or serum folate levels, and serum for reticulin antibodies. Five of nine patients selected for jejunal biopsy showed villous atrophy, and in some this seemed to be a true gluten-sensitive enteropathy.

Adult

Interpretation of serum and red cell folate results. A comparison of microbiological and radioisotopic methods.

Serum folate and red blood cell folate levels were measured in 167 patients using a microbiological method (Lactobacillus casei) and a commercial 125I radioassay kit. The results of the two methods were compared and related to the clinical findings to see which method more truly reflected the physiological folate status of the patient. There was a good linear correlation between the methods for both serum folate and red cell folate over the whole range encountered and the mean results obtained with each method were nearly identical. Both methods seemed to perform equally well in distinguishing low serum folate levels, but when the red cell folate results were compared to the clinical and other laboratory findings, the radioassay more closely related to the folate status of the patient. The study again emphasized the high number of low serum folate levels found in hospital patients and the importance of measuring the red cell folate.

Biological Assay

Cotrimoxazole and folate metabolism.

Cotrimoxazole 4 tablets daily (1 tablet = trimethoprim 80 mg and sulphamethoxzole 400 mg) was given for a period of six to fourteen days to 13 inpatients, and serum-folate levels were measured before and one day after the course of treatment. The results were compared with those from 8 patients not receiving antibacterial therapy, tested on admission and one week later. Two assay techniques were used, one employing Lactobacillus casei and the other 125I-labelled folate isotope dilution. The microbiological technique showed a significant decline in folic-acid levels in the serum after cotrimoxazole, and this decline was not seen in controls. By contrast, the radioisotope technique showed no significant alteration in serum-folate levels compared with controls. This suggests that cotrimoxazole does not depress true serum-folate and that many low microbiological results obtained during cotrimoxazole therapy reflect interference with the assay organism. There is insufficient evidence to incriminate cotrimoxazole as a significant cause of blood dyscrasias in excess of those which might occur on sulphonamide alone or even with other antibacterials.

Biological Assay

[Investigations of iron and folate levels in serum after implantation of heart valve prostheses (author's transl)].

In 90 patients with prosthetic heart valves, the presence of iron and folate deficiencies was assessed in a double-blind study. Anemia was found in 18 cases. Average serum iron levels were below the normal range and could be incremented significantly through the administration of 193 mg Fe++ daily during a 4-week period. No evidence of folate deficiency was found.

Anemia, Hypochromic

Serum folate and vitamin B12 levels in hypothyroid and hyperthyroid patients.

We measured the levels of serum folate and vitamin B12 in newly discovered hypothyroid (n =56) and hyperthyroid (n =47) patients and in age- and sex-matched control subjects (n =103). Except for one patient with latent pernicious anemia, serum folate and vitamin B12 levels did not differ greatly in our patients and in our control subjects. Another patient was receiving monthly injections of cyanocobalamin for previously diagnosed pernicious anemia. We conclude that abnormalities of thyroid function per se did not alter serum folate or vitamin B12 levels in our patients.

Adult

Folate-induced remission in aplastic anemia with familial defect of cellular folate uptake.

Severe aplastic anemia developed in a young man with an extensive family history of leukemia, pancytopenia, and neutropenia. Megaloblastic changes became evident, and treatment with high doses of folic acid resulted in striking clinical improvement. However, red-cell folate levels remained persistently low despite high serum folate levels. A defect in cellular folate uptake was suspected, and, indeed, uptake of 5-14CH3-H4-folate by stimulated lymphocytes and by bone-marrow cells from the patient was significantly reduced (P less than 0.05 as compared to normal cells. Further characterization of folate metabolism showed that intestinal absorption of the vitamin, membrane transport of 5-14CH3-H4-folate by mature red cells, folate utilization in the conversion of deoxyuridylate to thymidylate and polyglutamate formation were all normal. At least five other family members manifest decreased uptake of 5-14CH3-H4-folate by stimulated lymphocytes. These studies suggest that a genetically induced abnormality of folate uptake contributed to this patient's severe, but reversible, aplasia.

Adult

Neutrophilic hypersegmentation as an indicator of incipient folic acid deficiency.

The authors have identified a group of subjects with neutrophilic hypersegmentation who are normal or near-normal with respect to other hematologic indices (hemoglobin, mean corpuscular volume). In a high proportion of these subjects, serum folate levels are abnormally low. In this group and a non-hypersegmented-neutrophil control group there was a significant negative correlation between average numbers of neutrophilic lobes and serum folate levels. In the subjects with hypersegmented neutrophils the predominant alteration is a shift from three-lobed to five-lobed neutrophils. It is believed that neutrophilic hypersegmentation can be a valuable adjunct in documenting and/or uncovering incipient folate deficiency.

Erythrocytes

Dietary folate intake and concentration of folate in serum and erythrocytes in women using oral contraceptives.

Conflicting reports regarding the possible effect of oral contraceptives agents (OCA's) on folate status prompted us to evaluate the relationship between dietary folate intake and the concentration of folate in serum and erythrocytes among users and nonsuers of OCA's during two consecutive menstrual cycles. Twenty-two women (ages 19 to 28) had been on combination type OCA's for 4 months or more and a control group of 18 women (ages 18 to 29) had not used OCA's for at least 6 months prior to this study. The serum folate levels were lower in the OCA users than in the controls and the difference was statistically significant on day 5 of the menstrual cycle (P less t-an 0.05) but not on day 20. However, the differences in the erythrocyte folate levels and dietary folate intakes were not statistically significant between the two groups of subjects. There was a consistently higher degree of correlation between serum folate and folate intake among the control women than among the OCA users. Hematological parameters such as hemoglobin, hematocrit, mean corpuscular volume, mean corpusclar hemoglobin, mean corpuscular hemoglobin concentration and red cell count were similar in the two groups. It is concluded that the use of OCA's produces significantly lower serum folate levels during the first week of the menstrual cycle in spite of adequate folate intake.

Adult