Prenatal and postnatal diagnoses of thalassemias and hemoglobinopathies by HPLC

The conventional approach to qualitative and quantitative analyses of hemoglobin (Hb) molecules for the diagnoses of hemoglobinopathies requires a combination of tests. We used an automated HPLC (VARIANT) system to study alpha-thalassemia and beta-thalassemia syndromes in Thailand. The beta-thalassemia short program is applicable to the diagnosis of alpha-thalassemia and beta-thalassemia disorders, including Hb H, EA Bart's disease, and EF Bart's disease, in adults, newborns, and fetuses. The system cannot quantify accurately certain Hb molecules, such as Hb H and Hb Bart's. The alpha-thalassemia short program was therefore developed and used to quantify Hb Bart's to detect alpha-thalassemia genotypes in cord blood. This automated HPLC system is an alternative approach to the diagnosis of complicated thalassemia syndromes in Thailand and Southeast Asia.     

Technological properties,in vitro starch digestibility and in vivo glycaemic index of bread containing crude malva nut gum

The effects of crude malva nut gum (CMG) on the texture, digestion and the glycaemic index (GI) of white bread were investigated. Different CMG levels (0%, 2.5%, 5%, 7.5% and 10% w/w based on weight of flour) were added into a bread formula. The lightness (L*) and loaf‐specific volumes of the bread decreased with increased CMG levels. Firmness of the bread increased with increased CMG content. The in vitro starch digestion kinetics showed that bread containing CMG had generally lower predicted GI (74–81 vs. the control). In vivo human testing showed that increasing the CMG reduced the GI of the bread. The GI values of bread containing CMG ranged from 73 to 98 of the control. Overall liking scores of consumers ranged between ‘like slightly’ and ‘like moderately’ (6.1–7.1) compared to 7.1 for the control. Thus, the addition of CMG in carbohydrate‐based foods may lower the GI.     

Production,iron analysis and consumer perception of functional Thai Sinlek iron rice (Oryza sativa) drink

We produced the iron rice (IR) drink, determined the relevant chemical composition and antioxidant activity and tested the consumers’ degree of satisfaction. IR grains were digested with α-amylase and analysed chemical composition and antioxidant activity, while the hydrolysate (pH 4.0) and flavouring agents were added for final preparation of the IR drink. The IR hydrolysate contained a total iron content of 13.9 mg g⁻¹, a total phenolic content of 1.33 ± 0.07 mg gallic acid equivalent g⁻¹ and γ-oryzanol of 70.12 ± 2.7 mg kg⁻¹ rice grain, all of which inhibited the generation of free radicals in a concentration-dependent manner. The IR drink was composed of flavonoids and phenolic acids for which kaempferol was predominant. The members of all three groups of elderly subjects who consumed the low-dose IR (1.47 mg iron/100 mL), high-dose IR (2.74 mg iron/100 mL) and white rice (WR; 0.5 mg iron/100 mL) drinks expressed satisfaction and interest with the beverages at 91.2%, 100% and 88.9%, respectively. In terms of sensory evaluation, the low-dose IR drink revealed a higher acceptance rating than high-dose IR and WR (P < 0.05). Notably, the Sinlek rice drink was abundant with iron, anti-oxidative phenolics and γ-oryzanol, for which consumers expressed an overall degree of acceptance (8.1 ± 1.1).     

Increased serum levels of macrophage colony-stimulating factor (M-CSF) in alpha- and beta-thalassaemia syndromes: correlation with anaemia and monocyte activation

The Glasgow Coma Scale (GCS) is routinely used in the acute care setting after traumatic brain injury (TBI) to guide decisions in triage, based on its ability to predict morbidity and mortality. Although the GCS has been previously demonstrated to predict mortality, efficacy in prediction of functional outcome has not been established. The purpose of this study was to assess the value of the acute GCS in predicting functional outcome in survivors of TBI. This study used the Multicenter National Institute on Disability and Rehabilitation Research TBI Model Systems database of 501 patients who had received acute medical care and inpatient rehabilitation within a coordinated neurotrauma program for treatment of TBI. Initial and lowest 24 hr GCS scores were correlated with the following outcome measures: the Disability Rating Scale (DRS), Rancho Los Amigos Levels of Cognitive Functioning Scale (LCFS), and cognitive and motor components of the Functional Independence Measure (FIM(SM)-COG and FIM(SM)-M). Outcome data were collected at admission to and discharge from the inpatient TBI rehabilitation unit. Correlation analysis revealed only modest, but statistically significant, relationships between initial and lowest GCS scores and outcome variables. Initial and lowest GCS score comparison with outcome demonstrated the following correlation coefficients: admission DRS, -0.25 and -0.28; discharge DRS, -0.24 and -0.24; admission LCFS, 0.31 and 0.33; discharge LCFS, 0.27 and 0.25; admission FIM-COG, 0.36 and 0.37; discharge FIM-COG, 0.23 and 0.23; admission FIM-M, 0.31 and 0.31; discharge FIM-M, 0.25 and 0.21. The GCS as a single variable may have limited value as a predictor of functional outcome.     

Identification of Hb C [beta 6(A3)Glu-->Lys] in a Thai male

The propositus was a 29-year-old Thai male, whose electrophoretic pattern showed Hb A (58%) plus an abnormal hemoglobin (42%) with mobility identical to Hb A2 and Hb E. Protein sequencer analysis and tryptic peptide mapping of the beta chain indicated that the abnormal hemoglobin was Hb C [beta 6(A3)Glu-->Lys], rather than Hb E which is more commonly found in South East Asia. This conclusion was confirmed by direct sequence analysis of the propositus' DNA, which showed AAG as well as GAG at codon 6 of the beta gene, in agreement with heterozygosity for Hb C and Hb A. Furthermore, the beta gene framework (Ava II-, Bam HI+) of the propositus suggested that the beta C gene may have arisen from an independent mutation. Since Hb C and Hb E have the same mutation (Glu-->Lys) in the beta chain, although at different positions, and behave similarly in electrophoresis, cases of Hb C and Hb E may sometimes have been mistakenly identified for each other, based on whichever variant is most prevalent in the particular population.     

Thalassaemia classification by neural networks and genetic programming

This paper presents the use of a neural network and a decision tree, which is evolved by genetic programming (GP), in thalassaemia classification. The aim is to differentiate between thalassaemic patients, persons with thalassaemia trait and normal subjects by inspecting characteristics of red blood cells, reticulocytes and platelets. A structured representation on genetic algorithms for non-linear function fitting or STROGANOFF is the chosen architecture for genetic programming implementation. For comparison, multilayer perceptrons are explored in classification via a neural network. The classification results indicate that the performance of the GP-based decision tree is approximately equal to that of the multilayer perceptron with one hidden layer. But the multilayer perceptron with two hidden layers, which is proven to have the most suitable architecture among networks with different number of hidden layers, outperforms the GP-based decision tree. Nonetheless, the structure of the decision tree reveals that some input features have no effects on the classification performance. The results confirm that the classification accuracy of the multilayer perceptron with two hidden layers can still be maintained after the removal of the redundant input features. Detailed analysis of the classification errors of the multilayer perceptron with two hidden layers, in which a reduced feature set is used as the network input, is also included. The analysis reveals that the classification ambiguity and misclassification among persons with minor thalassaemia trait and normal subjects is the main cause of classification errors. These results suggest that a combination of a multilayer perceptron with a blood cell analysis may give rise to a guideline/hint for further investigation of thalassaemia classification.     

Erythroblast- and erythrocyte-bound antibodies in alpha and beta thalassaemia syndromes

Thirty-five Thai patients with various alpha-thalassaemia (alpha-thal 1/alpha-thal 2, alpha-thal 1/HbCS, HbCS/HbCS) and beta-thalassaemia (beta-thal/HbE, severe and mild form, HbE/HbE) syndromes were examined for the presence of immunoglobulins and C3d on o-tolidine positive erythroblasts in the bone marrow, and for the amounts of IgG of some specificities bound to circulating erythrocytes. In mild, but not in severe beta-thal/HbE and in alpha-thalassaemia, the percentages of Ig-positive erythroblasts were significantly higher than in controls and correlated well with the percentage of IgG-positive erythroblasts. By contrast, the percentages of IgM and C3d positive erythroblasts were low and similar in thalassaemic and control marrows. A substantial proportion of thalassaemic patients showed more erythrocyte-bound IgG than controls, but statistically significant elevations were seen only in severe beta-thal/HbE. Within a particular syndrome erythrocyte-bound IgG was more abundant in splenectomized than non-splenectomized subjects. It showed specificity for spectrin in some beta-thalassaemic patients and for band 3 protein in several individuals with alpha- or beta-thalassaemia. The results suggest that IgG antibodies play a role in the haemolysis of thalassaemia and that they are likely to be involved in the ineffective erythropoiesis in at least some of the syndromes studied.