Plasma homocysteine level and megaloblastic anemia status are two factors that can affect the quality of life of patients with multiple sclerosis (MS). We conducted this study to determine the effect of vitamin B12 and folic acid supplementation on serum homocysteine, megaloblastic anemia status and quality of life of patients with MS. A total of 50 patients with relapsing remitting multiple sclerosis (RRMS) included in this study which divided into 2 groups. The vitamin group received 5 mg folic acid tablet daily and 3 doses of vitamin B12 (1,000 mcg) injection and the other group received placebo and normal saline injection (same doses). The quality of life was measured by using Multiple Sclerosis Quality of Life-54 questionnaire (MSQOL-54). Fully automated fluorescence polarization immunoassay was used to measure serum homocysteine, vitamin B12 and folate. Complete blood count blood test was conducted to determine the anemia status. The mean homocysteine level reduced by 2.49 ± 0.39 µmol/L (p = 0.001), hemoglobin increased from 11.24 ± 1.54 to 13.12 ± 1.05 g/dL (p = 0.001), and mean corpuscular volume decreased from 95.50 ± 6.65 to 89.64 ± 4.24 in the vitamin group (p = 0.001). There was a significant improvement in the mental field of life quality in the placebo group (37.46 ± 19.01 to 50.98 ± 21.64; p = 0.001), whereas both physical and mental fields of quality of life were improved significantly in the vitamin group (40.38 ± 15.07 to 59.21 ± 12.32 and 29.58 ± 15.99 to 51.68 ± 18.22, respectively; p = 0.001). Serum homocysteine level decrease and anemia status improvement with vitamin B12 and folic acid supplementation reveal the potential role of these two vitamins in improving the life quality of MS patients. TRIAL REGISTRATION: Iranian Registry of Clinical Trials Identifier: IRCT2015100313678N7
Anemia* ; Anemia, Megaloblastic ; Blood Cell Count ; Erythrocyte Indices ; Fluorescence Polarization Immunoassay ; Folic Acid* ; Hematologic Tests ; Homocysteine* ; Humans ; Multiple Sclerosis* ; Multiple Sclerosis, Relapsing-Remitting ; Plasma ; Quality of Life* ; Vitamin B 12* ; Vitamins*

The term congenital adrenal hyperplasia (CAH) covers a group of autosomal recessive disorders caused by defects in one of the steroidogenic enzymes involved in the synthesis of cortisol or aldosterone from cholesterol in the adrenal glands. Approximately 95% of all CAH cases are caused by 21-hydroxylase deficiency encoded by the CYP21A2 gene. The disorder is categorized into classical forms, including the salt-wasting and the simple virilizing types, and nonclassical forms based on the severity of the disease. The severity of the clinical features varies according to the level of residual 21-hydroxylase activity. Newborn screening for CAH is performed in many countries to prevent salt-wasting crises in the neonatal period, to prevent male sex assignment in affected females, and to reduce long-term morbidities, such as short stature, gender confusion, and psychosexual disturbances. 17α-hydroxyprogesterone is a marker for 21-hydroxylase deficiency and is measured using a radioimmunoassay, an enzyme-linked immunosorbent assay, or a fluoroimmunoassay. Recently, liquid chromatography linked with tandem mass spectrometry was developed for rapid, highly specific, and sensitive analysis of multiple analytes. Urinary steroid analysis by gas chromatography mass spectrometry also provides qualitative and quantitative data on the excretion of steroid hormone metabolites. Molecular analysis of CYP21A2 is useful for genetic counseling, confirming diagnosis, and predicting prognoses. In conclusion, early detection using neonatal screening tests and treatment can prevent the worst outcomes of 21-hydroxylase deficiency.
Adrenal Glands ; Adrenal Hyperplasia, Congenital* ; Aldosterone ; Cholesterol ; Chromatography, Liquid ; Diagnosis ; Enzyme-Linked Immunosorbent Assay ; Female ; Fluoroimmunoassay ; Gas Chromatography-Mass Spectrometry ; Genetic Counseling ; Humans ; Hydrocortisone ; Infant, Newborn ; Male ; Mass Screening ; Neonatal Screening ; Prognosis ; Radioimmunoassay ; Steroid 21-Hydroxylase* ; Tandem Mass Spectrometry

OBJECTIVETo investigate the correlation of gestational age and birth weight with 17α-hydroxyprogesterone (17α-OHP) levels, and with results of adrenal hyperplasia newborn screening.

METHODUsing time-resolved fluorescence immunoassay, the authors measured concentrations of heel blood 17α-OHP by newborn dried blood spots on filter paper which included 29 hospitals newborns of Wujiang, Taicang, Zhangjiagang, Kunshan, and Suzhou, where there were 118 050 infants in total who had accurate gestational age and birth weight (62 490 males, 55 560 females). According to the classification by gestational age, there were 4 693 premature infants, 113 300 term infants and 57 overdue infants. According to the classification by birth weight, there were 4 172 infants with weight < 2 500 g, and 113 878 infants weight ≥ 2 500 g. And, in all premature infants, gestational age of 189 infants was < 32 weeks, 2 277 infants less than 36 weeks but ≥ 32 weeks, and 2 227 infants less than 37 weeks but not less than 36 weeks. Neonatal heel blood concentration of 17α-OHP was measured by dissociation enhanced lanthanide fluorescence immunoassay (DELFIA), and the correlation between 17α-OHP and gestational age or birth weight was retrospectively analyzed by using Spearman test.

RESULTThe distribution of 17α-OHP levels was skew. The 17α-OHP levels decreased significantly from very preterm births, moderately preterm, later period preterm to term infants [19.21 (8.07, 24.00), 12.35 (6.81, 18.00), 8.58 (5.66, 13.80), 5.60 (3.57, 8.51) , 3.34 (2.58, 5.23) nmol/L; 479.42, 62.25, 36.24, 23.30, 13.73 nmol/L;P all = 0.000]. The 17α-OHP levels decreases from very low birth weight (VLBW), extremely low birth weight (ELBW), low birth weight (LBW), normal birth weight to macrosomia [5.24 (3.24, 8.96) , 11.30 (6.84, 22.95) , 8.50 (5.28, 14.90) , 5.66 (3.61, 8.62) , 5.38 (3.40, 8.11) nmol/L; 485.26, 125.18, 39.50, 23.80, 22.15 nmol/L; P = 0.000 for all comparison]. Neonatal 17α-OHP levels and gestational age, body weight was significantly negatively correlated respectively -16.40 and -10.10 (P both = 0.000) by using Spearman test. Neonatal 17α-OHP levels and gestational age, body weight were binomially distributed, and the formulae were y = 0.105 5x²-2.457 6x + 17.689, R² = 0.980 3 and y = 0.411x²-3.988x+14.75, R² = 0.983. Little preterm infants, preterm infants and term infants in low birth weight infants 17α-OHP levels were significantly higher than non-low birth weight infants [11.20 (6.01, 18.90) vs 9.05 (5.85, 14.90) nmol/L, 9.76 (4.32, 10.35) vs 5.59 (3.56, 8.48) nmol/L, P all = 0.000].

CONCLUSIONNeonatal 17α-OHP levels and gestational age, body weight was significantly negatively correlated; in order to improve the accuracy and sensitivity, cut-off value of neonatal 17α-OHP should be adjusted according to gestational age and weight.

17-alpha-Hydroxyprogesterone ; blood ; Adrenal Hyperplasia, Congenital ; blood ; diagnosis ; Birth Weight ; Female ; Fluoroimmunoassay ; Gestational Age ; Humans ; Infant, Newborn ; Infant, Premature ; blood ; Infant, Very Low Birth Weight ; blood ; Male ; Neonatal Screening ; Retrospective Studies ; Sensitivity and Specificity

A fluorescence polarization immunoassay (FPIA) was developed for the analysis ofaflatoxins (AFs) using an anti-aflatoxin B1 (AFB1) monoclonal antibody and a novel fluorescein-labeled AFB1 tracer. The FPIA showed an IC50 value of 23.33 ng/mL with a limit of detection of 13.12 ng/mL for AFB1. The cross-reactivities of AFB1, AFB2, AFG1, AFG2, AFM1, and AFM2 with the antibody were 100%, 65.7%, 143%, 23.5%, 111.4%, and 2%, respectively. The group-specificity of anti-AFB1mAb indicated that the FPIA could potentially be used in a screening method for the detection of total AFs, albeit not AFG2 and AFM2. The total time required for analyzing 96 samples in one microplate was less than 5 min. This study demonstrates the potential usefulness of the FPIA as a rapid and simple technique for monitoring AFs.
Aflatoxins ; analysis ; Fluorescence Polarization Immunoassay

Animals ; Antibodies ; Estrone ; chemistry ; immunology ; Fluorescence Polarization Immunoassay ; methods ; Molecular Structure ; Rabbits ; Sensitivity and Specificity

OBJECTIVE: This study was conducted to confirm the results of the authors' previous research on schizophrenia manifesting high serum homocysteine and low folate levels. This study is anchored on a theory that a high serum homocysteine concentration affects schizophrenia by virtue of a neurotoxic mechanism, and on a report that some schizophrenia patients with high homocysteine levels benefited from high folate ingestion. METHODS: The serum homocysteine, folate, and vitamin B12 levels of 236 normal-control-group subjects and 234 schizophrenia subjects who met the diagnostic criteria based on DSM-IV-TR were compared. The homocysteine levels were measured via fluorescence polarization immunoassay, and the folate and vitamin B12 levels were determined via radioimmunoassay. RESULTS: The homocysteine levels of the patient group were significantly higher than those of the normal control group. The homocysteine level was more negatively correlated with the folate level in the schizophrenia group than in the control group. The percentages of female and male schizophrenia subjects manifesting high homocysteine levels were 33.8 and 51.5%, respectively. The percentage of schizophrenia subjects with low folate levels was 66.2%. In the low- and normal-folate-level groups, the patient group showed significantly higher homocysteine levels than the normal control group. The low-folate-level patient group particularly showed significantly higher homocysteine levels than the low-folate-level normal control group. CONCLUSION: Some schizophrenia patients with high serum homocysteine levels may have the genetic defect of having low folate serum levels. In such cases, folate ingestion may be a good management modality for clinical improvement.
Eating ; Female ; Fluorescence Polarization Immunoassay ; Folic Acid ; Homocysteine ; Humans ; Male ; Schizophrenia ; Virtues ; Vitamin B 12

Biomarkers ; blood ; Enzyme-Linked Immunosorbent Assay ; Fluoroimmunoassay ; Hepatitis B ; blood ; Hepatitis B virus ; Humans ; Luminescent Measurements

OBJECTIVETo prepare reference samples of Mycobacterium tuberculosis culture filtrate protein-10 (CFP-10) and CFP10-streptavidin fusion proteins (CFP10/SA) for time-resolved fluoroimmunoassay (TRFIA).

METHODSThe CFP10 gene was amplified by PCR from Mycobacterium tuberculosis strain H37Rv and cloned into pET24b, pET24b-streptavidin (SA) or pET21a-SA expression vectors. The recombinant proteins CFP10, CFP10-SA and SA-CFP10 were expressed in Rosetta cells, purified via nickel affinity chromatography and refolded by dialysis. The sensitivity and stability of the resultant proteins as reference samples were evaluated by double-antibody sandwich TRFIA.

RESULTSCFP10-SA and SA-CFP10 fusion proteins were expressed as inclusion bodies, whereas CFP10 was expressed in a soluble form. The resultant purity of the 3 recombinant proteins all exceeded 95%. TRFIA results showed that CFP-SA fusion protein possessed the best sensitivity (0.02 µg/L) and stability.

CONCLUSIONThe reference samples of CFP10 for TRFIA detection have been successfully prepared and can be used in the development of a diagnostic kit for Mycobacterium tuberculosis.

Bacterial Proteins ; genetics ; standards ; Fluoroimmunoassay ; methods ; Gene Amplification ; Mycobacterium tuberculosis ; isolation & purification ; Reference Standards

OBJECTIVETo develop a time-resolved fluoroimmunoassay (TRFIA) for detection of pertussis toxin (PT) S1 subunit for quality control of human PT vaccine.

METHODSA double antibody sandwich one-step method was used to establish the TRFIA for detecting PT S1 subunit in the vaccine.

RESULTSThe sensitivity of c peptide analysis reached 2.5 ng/ml without cross-reactions with other antigens. This assay could be used in detecting S1 subunit in the vaccine.

CONCLUSIONThe TRFIA for detecting PT S1 subunit is simple, sensitive and rapid for quality control of the PT vaccine.

Cross Reactions ; Fluoroimmunoassay ; methods ; Pertussis Toxin ; analysis ; Pertussis Vaccine ; chemistry ; standards ; Quality Control ; Sensitivity and Specificity

OBJECTIVETo study the expression of the ID3 protein in prostate cancer and its clinicopathological significance.

METHODSWe detected the expression of the ID3 protein in PC-3M cells by indirect immunofluorescence, and that in 29 prostate cancer and 15 prostate hyperplasia specimens by immunohistochemistry. Then we analyzed the correlation between the expression level of ID3 and the clinicopathological parameters.

RESULTSThe ID3 protein was expressed predominantly in the nucleus of PC-3M cells. Its expression rate was 82.7% (24/29) in the prostate cancer specimens, significantly higher than 6.6% (1/15) in prostate hyperplasia (P < 0.05), and was positively correlated with the Gleason score of prostate cancer (P < 0.05).

CONCLUSIONThe ID3 protein is expressed in prostate cancer, and is elevated with the increase of Gleason score.

Aged ; Aged, 80 and over ; Fluoroimmunoassay ; Humans ; Immunohistochemistry ; Inhibitor of Differentiation Proteins ; metabolism ; Male ; Middle Aged ; Neoplasm Proteins ; metabolism ; Prostatic Neoplasms ; metabolism ; pathology