Hemoglobin Oxidation in Stored Blood Accelerates Hemolysis and Oxidative Injury to Red Blood Cells

Introduction: Maintaining blood supply is a challenge in blood banks. Red blood cells stored at 4 °C have issues of biochemical changes due to metabolism of cells leading to biochemical changes, collectively referred to as “storage lesions”. Oxidation of the red cell membrane leading to lysis contributes to these storage lesions.

Methods: Blood bags with CPD-SAGM, stored at 4 °C for 28 days, were withdrawn aseptically on days 1, 14 and 28. Hematology analyzer was used to investigate RBC indices. Hemoglobin oxidation was studied by spectrophotometric scan of spectral change. RBC Lysis was studied by Drabkin’s assay, and morphological changes were observed by light and scan electron microscopy.

Results: Significant changes, RBC shows progressive changes in morphology echinocytes and spherocytes on day 28. There was 0.85% RBC lysis, ~20% decrease in percentage oxyhemoglobin, and a 14% increase in methemoglobin formation shows hemoglobin oxidation on day 28.

Conclusions: Oxidative damage to RBC with an increase in storage time was observed in the present study. The observed morphological changes to RBC during the course of increased time shows that there is progressive damage to RBC membrane and a decrease in hemoglobin concentration, and percentage RBC lysis is probably due to free hemoglobin and iron.


Biocompatibility and toxicity of novel iron chelator Starch-Deferoxamine (S-DFO) compared to zinc oxide nanoparticles to zebrafish embryo



Clinically approved iron chelators are effective in decreasing significant transfusional iron accumulation. Starch-Deferoxamine (S-DFO), a novel high molecular weight iron chelator, was produced to increase binding capacity to iron and reduce toxicity. Although its efficacy was established in one small cohort clinical trial, its potential adverse effect was not adequately addressed.


We utilized zebrafish model to assess S-DFO toxicity using following assays: mortality, teratogenicity, hatching rate, tail flicking, Acridine Orange staining for apoptosis detection, o-dianisidine staining for hemoglobin synthesis, and the level of Hsp70 as a general stress indicator. Embryos were exposed to different concentrations of S-DFO, Zinc Oxide nanoparticle (ZnO) (positive control), along with untreated control (UC).


S-DFO showed no significant mortality nor deformities at all tested concentrations (0.0–1000 μM). Thus, the LC50 is expected to >1000 μM. 100 μM S-DFO treatment did not affect embryo development (as judged by hatching rate); neuromuscular activity (as judged by tail flicking); and hemoglobin synthesis. Neither apoptosis, nor increase in Hsp70 level was noticed upon S-DFO treatment.


Our assays demonstrate that S-DFO does not induce cellular or biochemical stress and has no adverse effect on organ development of zebrafish embryos, suggesting its safe use as an iron chelator.https://www.sciencedirect.com/science/article/abs/pii/S0892036218301338

Genetic epidemiology of beta-thalassemia in the Maldives: 23 years of a beta-thalassemia screening program


The Maldives is an archipelago of 407,660 people according to population census of 2014, made up of 20 atolls, which has one of the highest prevalence of β-thalassemia worldwide. However, there is a dearth of studies related to β-thalassemia in the Maldives; therefore, in this study, we aimed to investigate the genetic epidemiology of β-thalassemia in Maldives. Blood samples were collected from 110,504 participants (1992–2015). Hemoglobin and RBC indices were measured on automated hematology analyzers. The quantitation of hemoglobin, HbA2, Hb F, and other abnormal Hb variants were assessed by HPLC. Molecular analysis was performed for the most common mutations in Southeast Asia for only 874 individuals either heterozygous or homozygous for these mutations using reverse dot blot hybridization. We screened 110,504 individuals for β-thalassemia between 1992 and 2015, which is ~ 30% of the entire population. The β-thalassemia carrier frequency was estimated to be 16.2%. Molecular diagnosis of 874 β-thalassemia carriers/major was performed for the most common seven mutations in Southeast Asia; of these, 139 patients were diagnosed as β-thalassemia major. This analysis showed that the most common mutations were IVS1 + 5G > C, (678; 77.6%), followed by the CD 30 (136; 15.6%). The least frequent mutation was FS8/9, (1, 0.001%), followed by IVS1 + 1G > T and CD15 (2; 0.2%). The frequency of β-thalassemia varies significantly among the 20 different atolls in Maldives. This study is expected to improve genetic counseling, creating awareness, enhance premarital screening, nd customize the prevention and treatment strategies based on the needs of each atoll. https://www.sciencedirect.com/science/article/abs/pii/S0378111920302134

Role of Zebrafish in medical research

In biomedical research always it is a challenge to find an experimental model to test various innovative drugs. In modern-day hematology, research zebrafish demonstrated its suitability and complex nature of known physiological functions of zebrafish has some resemblance to humans that makes these tiny fish in the forefront of cutting edge modern research. The blood of zebrafish is curious if we look at the RBC. In human RBC in peripheral blood is non-nucleated cells, but in these tiny fish blood has nucleated  RBCs. Zebrafish are commonly used as a vertebrate model to study multiple human diseases including cardiovascular disease. Zebrafish model is suitable for drug discovery research because its genome is homologous to the human genome by 70%. Besides, zebrafish develop rapidly during a short period where reaches to adulthood phase. In research zebrafish embryos and adult zebrafish are widely used to study metabolic and neurological functions.

Zebrafish larvae as a model to demonstrate secondary iron overload



Thalassemia is the most common genetically inherited blood disorder arising from a defect in hemoglobin production, resulting in ineffective erythropoiesis and severe hemolytic anemia. While transfusion therapy corrects the anemia, it gives rise to secondary iron overload. Current iron chelation therapy performed using deferoxamine, and the efficiency of this drug was demonstrated here using the zebrafish animal model.


Zebrafish larvae were exposed for 3 days to iron [100 μmol L−1 ferric ammonium citrate; 3‐6 days post fertilization (dpf)]. Then, iron treated larvae were exposed to 100 μmol L−1 deferoxamine for 3 days (6‐9 dpf). Total tissue iron concentration in the whole larvae, assessed by three different assays; inductively coupled plasma mass spectrometry, colorimetry (spectrophotometry), and microscopy using iron staining followed by imaging and quantification.


The three assays showed that iron treatment alone resulted in a significant increase in total iron. Deferoxamine treatment of the iron‐loaded zebrafish larvae showed a significant decrease in total iron concentration.


This study presented a clear evidence of the effectiveness of zebrafish larvae to use as a tool to study iron overload and open the door for studying the efficiency of potential new iron chelating compounds other than commercially available ones.https://onlinelibrary.wiley.com/doi/abs/10.1111/ejh.13035

Time Dependent Assessment of Morphological Changes: Leukodepleted Packed Red Blood Cells Stored in SAGM

Usually packed red blood cells (pRBCs) require specific conditions in storage procedures to ensure the maximum shelf life of up to 42 days in 2–6°C. However, molecular and biochemical consequences can affect the stored blood cells; these changes are collectively labeled as storage lesions. In this study, the effect of prolonged storage was assessed through investigating morphological changes and evaluating oxidative stress. Samples from leukodepleted pRBC in SAGM stored at 4°C for 42 days were withdrawn aseptically on day 0, day 14, day 28, and day 42. Morphological changes were observed using scanning electron microscopy and correlated with osmotic fragility and hematocrit. Oxidative injury was studied through assessing MDA level as a marker for lipid peroxidation. Osmotic fragility test showed that extended storage time caused increase in the osmotic fragility. The hematocrit increased by 6.6% from day 0 to day 42. The last 2 weeks show alteration in the morphology with the appearance of echinocytes and spherocytes. Storage lesions and morphological alterations appeared to affect RBCs during the storage period. Further studies should be performed to develop strategies that will aid in the improvement of stored pRBC quality and efficacy. http://www.hindawi.com/journals/bmri/2016/4529434/


Incidence of blood groups among blood donors in Maldives

Prevalence of blood groups among different ethnic groups have variation in different part of the world. There is no published data about the prevalence of blood groups in Maldives. The data shown here is the incidence trend among blood donors in blood bank at Maldivian Blood Services. As expected O Rh+ blood group is most common blood group among Maldivians.

Blood group

Maldives World “hot spot” for Thalassemia

Maldives is a world ‘hot spot’ for thalassemia. The prevalence of b-thalassemia in the population of this country is 18.1%; 1 in 5 people are carriers of the disease and 1 in 120 live births are estimated to be thalassemia majors. Currently, there are more than 700 thalassemia major patients who require regular blood transfusions, and as a result, 60% of the blood utilization of the Maldives treats these patients.

Cancer Awareness Program in Hulhmeedhoo

Cancer is alarmingly increasing in Hulhumeedho for the last decade the data shows.

Cancer awareness workshop held in Adducity Hulhumeedhoo on 22-23rd June 2012. The target audience was women of age group 21-45. The areas covered on this workshop were introduction of cancer, and common predisposing factors for cancer. Major areas covered among most common female cancers were cervical cancer, uterine and breast cancer.