Human serum albumin variants in China: a molecular epidemiological investigation and literature review

Background Bisalbuminemia is a hereditary and/or acquired abnormality characterized by a double albumin (ALB) band on serum protein electrophoresis. However, there have been no epidemiological investigations of ALB variants in Chinese populations. Methods This retrospective study examined 71,963 unrelated subjects from five provinces in southern China. ALB variants were screened by cellulose acetate electrophoresis at pH 8.6 and ALB mutations were confirmed by polymerase chain reaction-DNA sequencing. Results The average incidence of inherited bisalbuminemia in the southern Chinese population was 0.0264% (19/71,963). Thirteen cases showed slow and six showed fast genetic variants on cellulose acetate electrophoresis. Four kinds of ALB variants were identified: proalbumin Lille (p.Arg23His), ALB Castel di Sangro (p.Lys560Glu), ALB Fukuoka-1 (p.Asp587Asn), and a novel ALB Wuxi (p.Lys562Glu). The gene frequency of ALB variants in the Wuxi region (0.126%, 13/10,297) was significantly higher than in other regions in southern China, and 90.9% (10/11) of cases of proalbumin Lille were also found in the Wuxi region. Conclusions This study provides the first report of the detailed prevalence and molecular characterization of ALB variants in southern China. Compared with other areas of China, Wuxi had a different pattern of ALB variants and a high prevalence of proalbumin Lille.

Evaluation of Microchip-Based Point-Of-Care Device “Gazelle” for Diagnosis of Sickle Cell Disease in India

Sickle cell disease is a major public health problem in India. Lack of rapid and reliable diagnostic methods result in many avoidable deaths in affected population. Current diagnostic tools are laboratory based, expensive and need trained manpower. Here, we evaluated the performance of a microchip-based cellulose acetate electrophoresis test, “Gazelle” in the tribal-dominated Indian states of Chhattisgarh and Madhya Pradesh. A total of 1,050 patients were screened by sickle cell solubility, hemoglobin (cellulose acetate) electrophoresis, high-performance liquid chromatography (HPLC) and Gazelle. Of the total 1,027 test results obtained, 960 tests were “Valid” (93.5%) and included in the analysis. Gazelle identified all patients with disease (HbSS and Thalassemia Major) with 100% accuracy. Gazelle demonstrated 100% sensitivity when comparing sickle cell disease (SCD) vs. sickle cell trait and SCD vs. normal. Specificity was 98.9% and 99.5% when comparing SCD vs. trait and trait vs. normal, respectively. Specificity was 99.8% when comparing SCD vs. normal and sensitivity was 99.3% when comparing trait vs. normal. Overall, Gazelle yielded a high accuracy (99.0%) compared to reference standard tests (hemoglobin electrophoresis and HPLC). Gazelle is a low-cost, rapid diagnostic test with high accuracy for detecting SCD both quantitatively and qualitatively. Gazelle can be a potential screening tool for the rapid diagnosis in resource limited settings and developing countries with high burden of hemoglobin disorders.

Blood protein polymorphism and differentiation of selected indigenous cattle breeds in Nigeria

Genetic variation is the basis of effective improvement in farm animals. Population differentiation is used for objective choice of parental genotypes that constitutes new hybrids in crossbreeding. In Nigeria, population characteristics of selected indigenous cattle breeds have not been fully documented. Therefore, blood protein electrophoretic patterns of selected indigenous cattle breeds in Nigeria were assessed. Blood samples (5mL) were taken underneath the tail by venipuncture from 40 cattle randomly selected from each of the five breeds. The samples were subjected to cellulose acetate electrophoresis to determine the genetic variants of haemoglobin (Hb), carbonic anhydrase (CA) and transferrin (Tf) following standard procedure. Data were analysed using descriptive statistics, cluster analysis and Euclidean genetic distance. Allele frequencies ranged between 0.10 (Hb ) and 0.90 (Hb ), 0.11 (CA ) and 0.89 (CA ) and 0.02 (Tf ) and 0.49 (Tf ) across the breed.Two main clusters from the dendrogram were observed for each of Hb, CA and Tf. Euclidean genetic distance at the blood protein polymorphism level between WF and SG, WF and RB, WF and BK, WF and Muturu were 29, 30, 80 and 93, respectively. Genetic variants of transferring were largest within breed which indicated potential for selection

Evaluation of haemoglobin variants of Red Sokoto goats and Yankasa sheep in Beji, Niger State, Nigeria: Effect on body weight and reproductive parameters

Blood was sampled from 76 Red Sokoto (RS) goats and 76 Yankasa Sheep (YS) to evaluate Haemoglobin (Hb) genotypes and their effect on body weight and some reproductive parameters of the animals. The blood samples were analyzed by cellulose acetate electrophoresis. The electrophoretic test revealed the presence of three co-dominant alleles (A, B and C) which produced four haemoglobin genotypes (AA, AB, BB andBC) in RS goats, and two codominantalleles (AandB) which gave rise to two haemoglobin genotypes (AA and AB) in YS. The frequencies of alleles A, B and C were 0.32, 0.61 and 0.07 in RS goats; and 0.945 and 0.055 for alleles A and B in YS. The estimated heterozygosities were 0.406 and 0.118 in RS goats and YS while the estimated inbreeding coefficients were 0.009 and 0.008 in the RS goats and YS, respectively. The observed and expected genotype frequencies deviated significantly from the expected Hardy-Weinberg equilibrium (P 0.05;chi-square=7.82for RS goats, P 0.01;chi-square = 13.13 for YS).There were significant (P<0.05) differences observed in the body weight of male YS as a result of the Hb type; individuals with Hb AB were observed to have higher body weight (44.00 kg) compared to those with Hb AA (22.83 kg).No significant (P>0.05) difference was observed however in the body weight of RS goats. Male YS with Hb AB had longer (P 0.05) scrotum (26.00 kg) and wider scrotal circumference (33.00 kg) compared to those with Hb AA (18.83kg;25.67kg,respectively). No significant (P>0.05) difference was however observed in there productive parameters of RS goats. It was concluded that polymorphism exist at the Hb locus in the YS and RS goats sampled in Beji and this had an influence on body weight and the reproductive parameters evaluated particularly in the sheep. More heterozygotes were also observed in the population studied there by leading to alow rate of in breeding coefficient in the animals.

Diversity of West African dwarf goat in southwestern Nigeria based on allozyme Markers

Effective conservation, rational management and inadequate information on genetic diversity are the major challenges in livestock production. Genetic diversity has been used to reveal the extent of differentiation within livestock species. However, information on the use of allozymes in genetic diversity of the West African Dwarf (WAD) goat is insufficient. Therefore, genetic diversity of the WAD goat populations in southwestern Nigeria was investigated in this study. Three protein loci markers were used. Blood (5 mL each) samples were randomly collected from20, 20, 40 and 60 goats from Ondo ,Oyo, Ogun andOsun States respectively. The samples were subjected to cellulose acetate electrophoresis to determine the genetic variants at Haemoglobin (Hb), Carbonic Anhydrase (CA) and Transferrin (Tf) loci. Another set of blood (5 mL) from 20 different individual animals were randomly obtained from each of Ondo, Oyo, Ogun, and Osun States. Allele frequency, observed heterozygosity (H ), Polymorphic Information Content (PIC), F-statistic (F F and F ), gene flow (Nm), gene diversity (D), number of alleles per loci (A ), effective number of allele (AE) Mean Number of Allele (MNA) were generated from the data obtained. Data were analysed using Hardy-Weinberg equilibrium (HWE) at α0.05. The allele frequency ranged between 0.11 (Hb ) and 0.58 (Hb ), 0.17 (CA ) and 0.44 (CA ) and 0.08 (Tf ) to 0.60 (Tf ). Deviation from HWE was not significant in all populations except at Tf locus (0.00). The H ranged from 0.43 to 0.62 andNm and D ranged between 3.68 and 32.40 and 0.34 to 0.50 respectively. TheMNAwas 0.67 butA ranged from 1.52 to 2.00. The allozymes revealed some level of genetic diversity and a genetic differentiation indicative of the amount of genetic differences among individualswithin theWestAfricanDwarf goat population.

Production of sialic acid rich glycopeptide from bovine κ-casein glycomacropeptide by hydrolyzing with papain

Bovine κ-casein glycomacropeptide (GMP) is a sialic acid containing glycopeptide having many biological activities. The study described in this research communication was undertaken to determine whether sialic acid rich glycopeptide can be produced from GMP by proteinase treatment. A sample of GMP was hydrolyzed with papain, and the obtained hydrolysate was chromatographed on a column of diethylaminoethyl-Sephacel to obtain a glycopeptide fraction (GPF). This product accounted for average 48.1% dry weight of GMP or 81.1% total recovered sialic acid from GMP. The content of sialic acid (expressed as % dry weight) was 1.7 times higher in GPF (22.6) than in unhydrolyzed GMP (13.4). Major differences in amino acid composition between GPF and GMP were found in the contents (mol%) of: lysine (<1 and 4.5, respectively), serine (20.3 and 10.3, approximately twice higher in GPF), asparagine/aspartic acid and isoleucine. The contents of the last two amino acids were approximately twice lower in GPF. On gel filtration chromatography with Sephacryl S-100, GMP was eluted as a single peak with elution volume similar to that of dimeric β-lactoglobulin (36.6 kDa) whereas GPF was eluted in two peaks both with elution volumes greater than that of α-lactalbumin (14.2 kDa). These peak fractions containing high (fraction I) and low (fraction II) molecular size glycopeptides gave different sialic acid to peptide ratio, which was 1.7 times higher in fraction I than in fraction II. Results of size exclusion HPLC on Superdex-75 were consistent with those of gel filtlation chromatography. On cellulose acetate electrophoresis, the mobility of GPF relative to that of GMP as 1.0 was found to average 1.2, suggesting a higher negative charge density in GPF than in GMP. It was concluded that papain digestion of GMP is an efficient method to produce glycopeptide with high sialic acid content.

Hematological Profiles in Hemoglobinopathy Patients in South Western Nepal

INTRODUCTION: Homozygous inheritance of the hemoglobin results in sickle cell anemia (SCA), heterozygous inheritance results in sickle cell trait (SCT). Sickle cell anemia and β-thalasemia (β-TT) have been a major health threat for the tharu living in the South-Western Terai of Nepal. This study is carried out to apply and optimise the phenotypic method and haematological profile to characterise the SCT, SCA and β-TT from suspected tharu community dwellers. MATERIAL AND METHODS: We enrolled 100 suspected cases of hemoglobinopathies of 12-14 years children and ≥15 years adults tharu community dwellers from Dang, Kapilvastu, Nepalgunj, Rupandehi and Nawalparasi of South-Western Nepal from May 2018 to November 2018. Five mililitre of blood was collected in EDTA vial and transported to the laboratory maintaining cold chain. The hematological profile was recorded after investigations. The hemolysate from blood samples were subjected to phenotypic testing by adopting cellulose acetate electrophoresis at pH 8.6. RESULTS: Our result showed the commonest hemoglobinopathy was SCT (38%) followed by β-TT (21%) and SCA (5%). Males were more affected with SCA (60%) while that of females were most affected with β-TT (57.1%). Only 44.7% females were affected with SCT while 42.9% males were affected with β-TT. The significant difference in mean was observed in Hb level (p=0.0001), RBC (p=0.004), MCHC (p=0.015) and RDW (p=0.028) whereas the non-significant difference in mean was observed for glucose 6 phosphate dehydrogenase (G6PD) level (p=0.063) in hemoglobinopathy patients. CONCLUSION: Most cases develop severe type of anemia as shown by change in hematological parameters. This information could advocate for timely counselling before constellation of associated condition appeared in hemoglobinopathy patients.

Prevalence of Sickle Cell Disease and Other Haemoglobin Variants in Calabar, Cross River State, Nigeria

Background: Sickle cell disease (SCD) is the commonest genetic disorder worldwide with a global prevalence of 20-25 million. About 12-15 million affected persons are in Sub-Sahara Africa with Nigeria bearing the highest burden of people living with sickle cell disease. SCD is a disease characterized as an autosomal, recessive, heterogeneous, and a monogenetic disorder caused by an A-to-T point mutation in the β-globin gene responsible for the production of abnormal hemoglobin S (HbS), which polymerizes in the deoxygenated state and results in the sickling of erythrocytes. Haemoglobin variants are mutant forms of haemoglobin in a population usually occurring as a result of genetic changes in specific genes, or globins that causes change on alterations in the amino acid. They could affect the structure, behavior, the production rate and the stability of the specific gene. Well-known haemoglobin variants such as sick-cell anaemia are responsible for diseases and are considered haemoglobinopathies. Other variants cause no detectable pathology and are thus considered as non-pathological variants. Aim: The study is aimed at evaluating the burden of sickle cell disease and other haemoglobin variants in Calabar, South-South Nigeria. Methods: This is a retrospective study done at the haematology laboratory of University of Calabar Teaching Hospital, Calabar. Cellulose acetate electrophoresis at alkaline pH was used for the evaluation of haemoglobinopathies. The data were entered into Microsoft Excel 2016 spreadsheet and analysed with the IBM SPSS Version 22. Data were summarized into percentage of different phenotypes. Results: Results of the total 3648 haemoglobin electrophoresis recorded, 1368 (37.50%) were male while the remaining 2280 (62.5%) females given a male to female ratio of 1:1.7. Five haemoglobin phenotypes were identified as HbAA, HbAS, HbAC, HbSC and HbSS. The overall average values of their prevalence were HbAA 64.78%, HbAS 32.62%, HbSS 2.14%, HbAC 0.33%, HbSC 0.14%. Thus, the prevalence of SCD (Prevalence of HbSS+HbSC) was 2.28%. The highest proportion of SCD was observed in 2011 with least in 2016 and 2017 respectively. Conclusion: The prevalence of SCD and other haemoglobin variants in Calabar is similar to that of the national prevalence rate. There is need for continuous enlightenment and premarital counselling on the pattern of inheritance of SCD most especially with the increased burden of sickle traits in the environment has reported in this study.

Glucose 6 Phosphate Dehydrogenase Deficiency and Hemoglobinopathy in South Western Region Nepal: A Boon or Burden

Objectives: The study was carried out to optimize the phenotypic method to characterize the sickle cell trait (SCT), sickle cell anaemia (SCA) and β-thalassemia (β-TT) suspected sample from tharu community of South Western province-5, Nepal. SCT and SCA were further evaluated by genotypic method employing amplification refractory mutation system (ARMS PCR). Moreover, Glucose 6 Phosphate Dehydrogenase (G6PD) was estimated in those hemoglobinopathy to observe its prevalence. The accurate and reliable method can play an important role in reduction of morbidity and mortality rate. Results: The 100 suspected cases were subjected to phenotypic method adopting cellulose acetate electrophoresis and genotypic metod using ARMS PCR which portraits (5%) SCA positive test showing HBS/HBS, (38%) SCT positive trait HBA/HBS and (36%) cases normal HBA/HBA. β-TT (21%) cases were confirmed by electropherogram. G6PD deficiency was observed in (40%) of SCA, (18.4%) of SCT, (4.8%) of β-TT and (2.8%) in normal cases. Increased G6PD were developed only in SCT (5.3 %) and β-TT (4.8%). The study highlighted sickle cell disorder (SCD) and β-TT as the most common hemoglobinopathy coexisting with G6PD deficiency. Though hemoglobinopathy sometime could be protective in malaria but G6PD deficiency can cause massive hemolysis which may exacerbate the condition.