Nutritional anemia predominant form of anemia in educated young Thai women.

OBJECTIVE: Anaemia is one of the most common health problems worldwide, with a high prevalence in Africa and South East Asia, including Thailand. Thai women of childbearing age have an increased risk of anaemia due to several factors including underlying health problems, lifestyles and poor diet. Therefore, we investigated the prevalence of anaemia among female students of Chulalongkorn University (aged 18-22) and categorized causes of the anaemia. DESIGN: 400 Thai female student-volunteers, without known underlying diseases were subjected to blood tests; complete blood count, Haemoglobin typing and serum ferritin level. Bloods, having haemoglobin under 12 g/dl and hematocrit under 36%, were designated as anaemia. Then causes of anaemia are categorized into 3 groups; Iron deficiency, Thalassemia and Others. RESULTS: We found that 21% of the volunteers were anaemic. In 85 anaemic volunteers, they were classified as Iron deficiency anaemia (IDA); with low serum ferritin levels 42.4%, Thalassemia; total of 6 types 25.9%, IDA and Thalassemia 2.3% and Others 29.4% in which haemoglobin typing and serum ferritin level were normal. CONCLUSION: Iron deficiency anaemia (IDA) is the major cause of anaemia in Thai female students in our study. Several students were gradually developing anaemia where their haematocrit (Hct) and haemaglobin (Hb) were within reference range but mean corpuscular volume (MCV), mean corpuscular haemaglobin (MCH) and serum ferritin fell below reference range, indicating latent iron deficiency. A few volunteers had both IDA and Thalassemia and also Thalassemia with iron overloaded where health can be deteriorated without knowledge of having these conditions or proper health care. To improve their health, universities or public organizations should provide education and/or screen for anaemia. With the knowledge and understanding of their health issues or underling diseases, students themselves can prevent serious health conditions, improve university performances, and improve their quality of life.

Anti-Leukemic Effects on a U937 Cell Line of Fresh and Steamed Chinese Kale Juice and Their Pro-Apoptotic Effects via a Caspase-Dependent Pathway.

Chinese kale is a vegetable belonging to the family Brassicaceae in which members of this family produce unique metabolites called glucosinolates and isothiocyanates. These substances have been found to exhibit many benefits to human health. This study aimed to investigate and compare the contents of glucosinolates and isothiocyanates, and the anti-leukemic activity of fresh and steamed Chinese kale juice (CKJ). Cell viability and proliferation activity of U937 cells treated with CKJ were determined. Cell apoptosis and alterations of apoptosis-related protein expression were studied. Results showed that CKJ significantly decreased the viability of leukemic cells and inhibited cell proliferation in a dose- and time-dependent manner. After treatment with 5% v/v fresh and steamed CKJ for 24 h, the percentage of apoptotic cells increased to 53% and 36%, respectively. Increased amounts of cleaved caspase-3 in U937 cells treated with CKJ were observed, indicating that CKJ can trigger apoptotic cell death through a caspase-dependent pathway. Fresh CKJ was found to be more effective than steamed CKJ in suppressing cell survival and inducing cell apoptosis. The results suggest that Chinese kale possesses an anti-leukemic potential and could be further developed for cancer therapy and prevention. However, thermal cooking could reduce its beneficial function.

Exploring Matrix Effects on Binding Properties and Characterization of Cotinine Molecularly Imprinted Polymer on Paper-Based Scaffold.

Commercially available sorbent materials for solid-phase extraction are widely used in analytical laboratories. However, non-selective binding is a major obstacle for sample analysis. To overcome this problem, molecularly imprinted polymers (MIPs) were used as selective adsorbent materials prior to determining target analysts. In this study, the use of non-covalent molecularly imprinted polymers (MIPs) for cotinine adsorption on a paper-based scaffold was studied. Fiberglass paper was used as a paper scaffold for cotinine-selective MIP adsorption with the use of 0.5% agarose gel. The effects of salt, pH, sample matrix, and solvent on the cotinine adsorption and extraction process were investigated. Under optimal conditions, the adsorption isotherm of synthesized MIPs increased to 125.41 µg/g, whereas the maximum adsorption isotherm of non-imprinted polymers (NIPs) was stable at 42.86 µg/g. The ability of the MIP paper scaffold to absorb cotinine in water medium was approximately 1.8⁻2.8-fold higher than that of the NIP scaffold. From Scatchard analysis, two dissociation constants of MIPs were calculated to be 2.56 and 27.03 µM. Nicotine, myosmine, and N-nitrosonornicotine were used for selectivity testing, and the calculated selectivity factor of cotinine to nicotine, myosmine, and N-nitrosonornicotine was 1.56, 2.69, and 2.05, respectively. Overall, the MIP paper scaffold is promising for simple onsite sampling of cotinine and can be used to assess tobacco smoke exposure.

Production of the α and ß Subunits of Spirulina Allophycocyanin and C-Phycocyanin in Escherichia coli : A Comparative Study of Their Antioxidant Activities.

Allophycocyanin and c-phycocyanin have been reported to be potent antioxidants. In this work, the genes encoding the apo-proteins of allophycocyanin α (ApcA), allophycocyanin ß (ApcB), c-phycocyanin α (CpcA), and c-phycocyanin ß (CpcB) from Spirulina platensis were cloned, and the recombinant proteins were produced in Escherichia coli to study their antioxidant effects. All four recombinant phycocyanins could be produced in the soluble form and purified to more than 97% purity. The results of radical scavenging assays showed that the Trolox equivalent values for peroxyl radical scavenging by the ApcA, ApcB, CpcA, and CpcB proteins were 1.81 ± 0.2 µM, 1.98 ± 0.22 µM, 0.95 ± 0.15 µM, and 1.49 ± 0.15 µM, respectively. The IC50 values for hydroxyl radical scavenging of ApcA, ApcB, CpcA, CpcB, and Trolox were 269 ± 9 µg/mL, 190 ± 5 µg/mL, 129 ± 8 µg/mL, 108 ± 4 µg/mL, and 195 ± 12 µg/mL, respectively. These results indicated that allophycocyanin exhibited higher activity than c-phycocyanin in scavenging peroxyl radicals, whereas c-phycocyanin exhibited higher activity than allophycocyanin in scavenging hydroxyl radicals. All of the apo-phycocyanin subunits possessed strong antioxidant activities and can be further developed and applied to the food and drug industries. However, the selection of the most useful antioxidant should depend on the type of targeted free radical to obtain the highest efficiency.

Anti-oxidant activity of holo- and apo-c-phycocyanin and their protective effects on human erythrocytes.

This study was conducted to investigate the anti-oxidant activity of the recombinant apo-c-phycocyanin (c-PC) ß-subunit compared to native c-PC purified from Spirulina sp. The gene encoding the ß-subunit of c-PC was successfully cloned and expressed in Escherichia coli. The anti-oxidant capacities of recombinant apo-c-PC(ß) and native c-PC were evaluated by measuring their Trolox equivalent antioxidant capacities and examining their protective effects on erythrocytes from normal and homozygous haemoglobin E individuals against peroxyl radicals and hydrogen peroxide. The results demonstrated that the anti-oxidant capacities are native c-PC≫Trolox>recombinant apo-c-PC(ß). Both anti-oxidant proteins can potentially protect erythrocytes from oxidative damage. Expression of c-PC in bacteria reduces the cost and time for protein production, and the recombinant protein could be further developed to obtain a more efficient protein for therapeutic purposes.

Chimeric antibody-binding Vitreoscilla hemoglobin (VHb) mediates redox-catalysis reaction: new insight into the functional role of VHb.

Experimentation was initiated to explore insight into the redox-catalysis reaction derived from the heme prosthetic group of chimeric Vitreoscilla hemoglobin (VHb). Two chimeric genes encoding chimeric VHbs harboring one and two consecutive sequences of Fc-binding motif (Z-domain) were successfully constructed and expressed in E. coli strain TG1. The chimeric ZVHb and ZZVHb were purified to a high purity of more than 95% using IgG-Sepharose affinity chromatography. From surface plasmon resonance, binding affinity constants of the chimeric ZVHb and ZZVHb to human IgG were 9.7 x 10(7) and 49.1 x 10(7) per molar, respectively. More importantly, the chimeric VHbs exhibited a peroxidase-like activity determined by activity staining on native PAGE and dot blotting. Effects of pH, salt, buffer system, level of peroxidase substrate and chromogen substrate were determined in order to maximize the catalytic reaction. From our findings, the chimeric VHbs displayed their maximum peroxidase-like activity at the neutral pH (approximately 7.0) in the presence of high concentration (20-40 mM) of hydrogen peroxide. Under such conditions, the detection limit derived from the calibration curve was at 250 ng for the chimeric VHbs, which was approximately 5-fold higher than that of the horseradish peroxidase. These findings reveal the novel functional role of Vitreoscilla hemoglobin indicating a high trend of feasibility for further biotechnological and medical applications.

Co-expression of zinc binding motif and GFP as a cellular indicator of metal ions mobility.

A significant role of zinc-binding motifs on metal mobility in Escherichia coli was explored using a chimeric metal-binding green fluorescent protein (GFP) as an intracellular zinc indicator. Investigation was initiated by co-transformation and co-expression of two chimeric genes encoding the chimeric GFP carrying hexahistidine (His6GFP) and the zinc-binding motif fused to outer membrane protein A (OmpA) in E. coli strain TG1. The presence of these two genes was confirmed by restriction endonucleases analysis. Co-expression of the two recombinant proteins exhibited cellular fluorescence activity and enhanced metal-binding capability of the engineered cells. Incorporation of the zinc-binding motif onto the membrane resulted in 60-fold more binding capability to zinc ions than those of the control cells. The high affinity to metal ions of the bacterial surface influenced influx of metal ions to the cells. This may affect the essential ions for triggering important cell metabolism. A declining of fluorescent intensity of GFP has been detected on the cell expressed of zinc binding motif. Meanwhile, balancing of metal homeostasis due to the presence of cytoplasmic chimeric His6GFP enhanced the fluorescent emission. These findings provide the first evidence of real-time monitoring of intracellular mobility of zinc by autofluorescent proteins.