Childhood aplastic anaemia with paroxysmal nocturnal haemoglobinuria clones: A retrospective single-centre study in South Africa

Background: Paroxysmal nocturnal haemoglobinuria (PNH) clones in children are rare but commonly associated with aplastic anaemia (AA) and myelodysplasia.Objective: This study aimed to determine the prevalence of PNH clones in paediatric patients with idiopathic AA, identify differences in clinical and laboratory features and outcomes, and determine the impact of clone size on clinical presentation.Methods: Patients with confirmed idiopathic AA who were tested for PNH between September 2013 and January 2018 at the Inkosi Albert Luthuli Central Hospital, Durban, KwaZulu-Natal, South Africa, were included. PNH clones were detected in neutrophils and monocytes by flow cytometry using fluorescent aerolysin, CD24, CD66b and CD14. Results: Twenty-nine children with AA were identified and 11 were excluded. Ten patients (10/18, 55.6%) had PNH clones ranging from 0.11% to 24%. Compared to the PNH-negative group, these children were older (median: 10 years vs 4 years, p= 0.02) and had significantly lower total white cell counts (median 1.7 × 109/L vs 3.2 × 109/L; p= 0.04). There was no difference in median absolute neutrophil count or haemoglobin concentration. Four patients in each group received immunosuppressive therapy (IST). At six months, all four patients with PNH clones had responded, compared to one in the PNH-negative group. Conclusion: More than half of children with AA had a PNH clone. The size of the clone did not impact clinical severity; however, IST use may positively impact prognosis. We recommend early initiation of IST in patients with AA to avoid delays associated with human leukocyte antigen typing.

Clinical features of five children with hepatitis-associated aplastic anaemia / 中国小儿急救医学

Objective:To investigate the clinical characteristics of hepatitis-associated aplastic anaemia(HAAA)in children.Methods:A retrospective analysis was performed on the clinical manifestations, laboratory examinations, treatments and other clinical data of five children with aplastic anemia(AA)diagnosed by bone marrow examination after admission with acute liver dysfunction admitted to the Department of Gastroenterology, Children′s Hospital Affiliated to Capital Institute of Pediatrics from January 2016 to December 2020.Results:All five children were boys and the onset age of these children ranged from 2 to 13 years.All of the five cases were acute onset and presented with jaundice.The time frame of the diagnosis of HAAA was 0 to 12 weeks from the presentation of the liver disease.One patient had simultaneous onset of hepatitis and aplastic anemia.The liver function was significantly improved at the diagnosis of HAAA in three patients and worsen in one patient.Only one patient showed CMV-DNA positive and the pathogen results of other patients were negative.Lymphocyte immunity disorders were found in all five patients, and the proportion of inhibitory/cytotoxic T lymphocytes(CD3 + CD8 + ) increased.Two children received hematopoietic stem cell transplantation, of which one died and one improved after transplantation.One child improved after treated with antithymocyteglobulin and cyclosporin.One child died due to severe infection.There was no significant improvement in one child treated with cyclosporine. Conclusion:HAAA should be alerted in acute hepatitis patients.Blood routine should be monitored even if liver function improves.Bone marrow tests are needed if patients have peripheral cytopenia in two or more lineages.Early and timely treatments with immunosuppressive therapy and hematopoietic stem cell transplantation can improve the prognosis.

Study of etiological spectrum and clinical profile of patients admitted in tertiary care hospital of South Gujarat, India

Background: Pancytopenia is defined as reduction of all three formed elements of blood below normal reference range. The symptoms are fatigue, fever, dizziness and weight loss. Evaluation was done using complete hemogram and peripheral smear. The presenting marrow biopsy is most useful and accurate in evaluation of pancytopenia.Methods: Two ml of anticoagulated blood was collected for complete hemogram. The peripheral blood smear was stained with Leishman's stain and studied. Bone marrow biopsy and aspiration was done in all the patients to identify etiology.Results: Predominance was seen in the age group of 31-60 years. Most common cause of pancytopenia was megaloblastic anaemia in this study compared to other studies all over the world where most common cause was aplastic anaemia. This reflects higher prevalence of megaloblastic anaemia in the Indian subjects.Conclusions: A comprehensive clinical, haematological land bone marrow study of patients with pancytopenia usually helps in identification of the underlying cause.

Anti-thymocyte globulin in haematology: Recent developments

Anti-thymocyte globulin (ATG) is a polyclonal antiserum introduced into clinical medicine more than 30 years ago. It induces a broad non-specific immunosuppression. In haematology, standard indications are severe aplastic anaemia and prophylaxis and treatment of graft-versus-host disease (GVHD) (after allogeneic transplantation). For aplastic anaemia, ATG from horses has been found to be superior to ATG from rabbits. In the situation of allogeneic transplantation, ATG lessens the risk of chronic GVHD but may not improve survival. There is current controversy regarding which patients benefit most from ATG and what the ideal dosage is. It is likely that in the coming years a more specific immunosuppressive will be developed that will minimize GVHD while maintaining the graft-versus-malignancy effect.

A prospective clinico‑hematological study in 100 cases of Pancytopenia in a tertiary care teaching hospital

Background: Pancytopenia is encountered regularly in haematology practice, yet there exist only few published assessments of the frequencies of various aetiologies and this exhibit substantial geographic variation. Pancytopenia is a manifestation of many life- threatening diseases with a wide range of differential diagnosis. Haematological investigation forms the bedrock in the detection and management of patients with pancytopenia.Methods: This study is a prospective study conducted in the Institute of pathology and haematology, Madras medical college and Rajiv Gandhi Government General Hospital, Chennai during the period from August 2015 to August 2016 on 100 cases. Case selection is based on clinical features and supported by laboratory evidence. Peripheral smear was obtained and stained by Leishman stain for all cases and examined in detail. Bone marrow aspiration /biopsy was subsequently carried out under aseptic precautions.Results: Among the 100 cases studied, age of the patients ranged from 13 to 80 years with a slight male predominance. Most of the patients presented with generalized weakness and fever. The commonest cause for pancytopenia was aplastic anaemia followed by megaloblastic anaemia. The other causes include acute myeloid leukaemia, myelodysplastic syndrome, myelofibrosis, multiple myeloma, malarial parasite, miliary tuberculosis and osteopetrosis.Conclusions: Pancytopenia can be diagnosed, and its etiological profile can be ascertained with the help of detailed clinical history, meticulous physical examination and haematological investigations. Every attempt should be done to establish the underlying cause so that treatable conditions are diagnosed without delay and prognosis is improved.

To evaluate the clinical and etiological profile of patients presenting with pancytopenia in Government Dharmapuri Medical College Hospital, Dharmapuri

Introduction: The frequency of underlying pathology causing pancytopenia varies considerably depending upon various factors including geographic distribution and genetic disturbances. The severity of pancytopenia and the underlying pathology determine the management and prognosis of the patients with pancytopenia. The basic investigations in a suspected case of pancytopenia include Complete Blood Count with peripheral blood film and Reticulocyte count. Aim: To evaluate the clinical and etiological profile of patients presenting with pancytopenia. Materials and methods: A total of 65 patients were identified over a period of 12months were included in the study. Basic investigations were performed for each patient including Haemoglobin, Total leukocyte count, Platelet count, Reticulocyte count. Absolute values including packed cell volume (MCV), mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC) were calculated for every patient. Results: 24.62% of patients had aplastic anemia. Of which 10 (62.5%) were male and 6 (37.5%) were female Mean age of patients with aplastic anemia is 33.43 yrs. Average Hb% of these patients are 4.78 gms%.56.25% of patients had thrombocytopenia < 100000 cells/chum Megaloblastic anaemia was more common in females (14) compared to males (11) in our study. While aplastic anemia is common in males (10) than females (6); both of which are statistically significant. Anisopoikilocytosis (92%) and hyper segmented neutrophils (92%) are the most common findings in peripheral smear of patients with megaloblastic anaemia. Hypersplenism, MDS, viral infections were P. S. Rani, K. Sureshkumar. To evaluate the clinical and etiological profile of patients presenting with pancytopenia in Government Dharmapuri Medical College Hospital, Dharmapuri. IAIM, 2017; 4(6): 125-131. Page 126 common in males; while acute leukaemia’s, myelofibrosis were common in females. Both of which is statistically insignificant. Conclusion: As much physicians should have a high index of suspicion for Vitamin B12 deficiency when dealing with patients presenting with symptoms of anemia such as pallor and weakness and/or diagnosed with pancytopenia on further workup. The finding of hyper segmented neutrophils in the peripheral smear will guide in the diagnosis of megaloblatic anemia.

Treatment for the relapsed or refractory severe aplastic anaemia / 国际儿科学杂志

Severe aplastic anaemia is a marrow failure syndrome characterized by pancytopenia and hypocellular bone marrow.Immunosuppressive therapy(IST)consisting of antithymocyte or antilymphocyte globulin (ATG) and cyclosporine has been recommended for children lacking HLA-matched sibling donor.For children unresponsive to initial immunosuppressive therapy or those who relapsed after first immunosuppressive therapy,they can choose repeat course of immunosuppressive therapy,allogeneic hernatopoietic stem cell transplantation,new drugs or clinical trials.And the prognosis has been greatly improved in recent years,dueing to the development of allogeneic hematopoietic stem cell transplantation and the application of new drugs such as eltrombopag.This article reviews researches advance in different therapies for children with relapsed or refractory severe aplastic anaemia,aimed at guaiding treatment methods.

Detecção de células com fenótipo HPN por citometria de fluxo, utilizando análise multiparamétrica / Detection of PNH cells by flow cytometry, using multiparameter analysis

Introduction: The laboratory diagnosis of paroxysmal nocturnal hemoglobinuria (PNH), disease that is categorized by reduced synthesis of glycosylphosphatidylinositol (GPI) anchor, is based on the detection of blood cells deficient in GPI-anchored proteins by flow cytometry. PNH clones have been detected in patients with aplastic anaemia (AA) and myelodysplastic syndrome (MDS), with therapeutic implications. Objectives: To validate a sensitive assay for detection of GPI-anchored protein-deficient cells, by flow cytometry, and to analyze the clone frequency in AA and MDS patients. Methods: Samples from 20 AA patients, 30 MDS patients and 20 adult volunteers (control group) were analyzed using monoclonal antibodies to CD16, CD24, CD55 and CD59 (neutrophils); CD14 and CD55 (monocytes); CD55 and CD59 (erythrocytes); besides fluorescent aerolysin reagent (FLAER) (neutrophils and monocytes) and lineage markers. The proportions of PNH cells detected in neutrophils and monocytes, using different reagent combinations, were compared by analysis of variance (ANOVA) and Pearson's correlation. Results: PNH cells were detected in five (25%) AA patients, and the proportions of PNH cells varied from 0.14% to 94.84% of the analyzed events. PNH cells were not detected in the MDS patients. However, by the analysis of these samples, it was possible to identify the technical challenges caused by the presence of immature and dysplastic circulating cells. FLAER showed clear distinction of GPI-deficient cells. Conclusion: Multiparameter flow cytometry analysis offers high sensitivity and accuracy in the detection of subclinical PNH clones. FLAER shows excellent performance in detection of PNH neutrophils and monocytes...

Introdução: O diagnóstico laboratorial da hemoglobinúria paroxística noturna (HPN), doença caracterizada por deficiência de síntese da molécula de ancoragem glicosilfosfatidilinositol (GPI), baseia-se na detecção de células sanguíneas deficientes em proteínas ancoradas ao GPI, por citometria de fluxo. Clones de células com fenótipo HPN podem ser detectados em pacientes com anemia aplásica (AA) e síndrome mielodisplásica (SMD), com implicações terapêuticas. Objetivos: Validar técnica sensível para detecção de células HPN, por citometria de fluxo, e avaliar a frequência dos clones em pacientes com AA e SMD. Métodos: Amostras de 20 pacientes com AA, 30 pacientes com SMD e 20 voluntários (controles) foram analisadas, utilizando anticorpos monoclonais anti-CD16, CD24, CD55 e CD59 (neutrófilos); CD14 e CD55 (monócitos); e CD55 e CD59 (hemácias); além do reagente de aerolisina fluorescente (FLAER) (neutrófilos e monócitos) e marcadores de linhagem celular. A comparação do tamanho dos clones HPN detectados em neutrófilos e monócitos, pelas diferentes combinações de reagentes, foi realizada por análise de variância (ANOVA) e correlação de Pearson. Resultados: Em cinco (25%) pacientes com AA foram identificadas células HPN, em proporções entre 0,14% e 94,84% dos eventos analisados. O clone não foi detectado nos pacientes com SMD. Contudo, a análise dessas amostras permitiu evidenciar as dificuldades técnicas secundárias à presença de células imaturas e displásicas circulantes. O reagente FLAER propiciou separação precisa das células alteradas. Conclusão: A análise multiparamétrica por citometria de fluxo apresenta sensibilidade...

Cytogenetic profile of aplastic anaemia in Indian children.

Background & objectives: Aplastic anaemia is a rare haematological disorder characterized by pancytopenia with a hypocellular bone marrow. It may be inherited/genetic or acquired. Acquired aplastic anaemia has been linked to many drugs, chemicals and viruses. Cytogenetic abnormalities have been reported infrequently with acquired aplastic anaemia. Majority of the studies are in adult patients from the West. We report here cytogenetic studies on paediatric patients with acquired aplastic anaemia seen in a tertiary care hospital in north India. Methods: Patients (n=71, age 4-14 yr) were diagnosed according to the guidelines of International Agranulocytosis and Aplastic Anaemia Study. Conventional cytogenetics with Giemsa Trypsin Giemsa (GTG) banding was performed. Karyotyping was done according to the International System for Human Cytogenetics Nomenclature (ISCN). Results: Of the 71 patients, 42 had successful karyotyping where median age was 9 yr; of these 42, 27 (64.3%) patients had severe, nine (21.4%) had very severe and six (14.3%) had non severe aplastic anaemia. Five patients had karyotypic abnormalities with trisomy 12 (1), trisomy 8 (1) and monosomy 7 (1). Two patients had non numerical abnormalities with del 7 q - and t (5:12) in one each. Twenty nine patients had uninformative results. There was no difference in the clinical and haematological profile of patients with normal versus abnormal cytogenetics although the number of patients was small in the two groups. Interpretation & conclusions: Five (11.9%) patients with acquired aplastic anaemia had chromosomal abnormalities. Trisomy was found to be the commonest abnormality. Cytogenetic abnormalities may be significant in acquired aplastic anaemia although further studies on a large sample are required to confirm the findings.

A novel autologous stem cell procedure for the treatment of aplastic anaemia using reprogrammed mature adult cells: a pilot study.

Background & objectives: Aplastic anaemia is a life threatening rare bone marrow failure disorder. The underlying haematopoietic cellular deficit leads to haemorrhage, infection and severe anaemia. The treatment of choice for this haematological condition is allogeneic bone marrow transplantation from fully matched HLA sibling. Though this procedure is curative in the majority of young patients with aplastic anaemia, extending this benefit to older patients or those lacking a family donor remains a major challenge. Herein, the safety and efficacy of infusing autologous retrodifferentiated haematopoietic stem cells (RHSC) into four patients with aplastic anaemia without the use of any pre- or post-conditioning regimen including immunosuppression is described. Methods: Un-mobilized, mononuclear cells were harvested from four patients with acquired aplastic anaemia by aphaeresis. Mononuclear cells of patients were cultured with purified monoclonal antibody against the monomorphic regions of the beta chain of MHC class II antigens (Clone CR3/43) for 3 h, to obtain autologous RHSC. Autologous RHSC were washed and infused into the four patients without the use of any pre- or post-conditioning regimen. Thereafter, the efficacy (engraftment) of autologous RHSC was assessed in these patients. Results: Following single infusion of the autologous RHSC, two of the four patients with aplastic anaemia become transfusion independent for more than seven years. Karyotyping and G-banding analysis prior and post-procedure in all patients remained the same. Interpretation & conclusions: The findings of this pilot study demonstrated the functional utility of reprogrammed fully differentiated adult cells into pluripotent stem cells with extensive repopulation potentials in a human setting and without any pre- or post-conditioning regimen, including immunosuppression. This autologous approach of stem cell creation may broaden the curative potentials of stem cell therapy to a wider population of patients with aplastic anaemia, including many patients suffering from other haematological and non-haematological disorders.

Carbimazole-induced aplastic anaemia - a case report

Antithyroid drugs have been used for more than 50 years for the management of hyperthyroidism. Most patients tolerate treatment well, but some may develop rare life threatening side effects such as agranulocytosis and aplastic anaemia. Clinical experience with the latter condition is extremely limited. We report on a case of carbimazole-induced aplastic anaemia caused by hypocellular bone marrow and associated plasmacytosis in a thyrotoxic patient chronically treated with carbimazole. This resolved after substitution with propylthiouracil. The clinical course was complicated by neutropaenic septicaemia and atrial fibrillation.

Study of the Therapeutic Effects of Bushenshengsui I on Mice With Experimental Aplastic Anaemia / 中国药房

OBJECTIVE:To study the possible mechanism of Bushenshengsui I(BSSS I),a kind of compound Chinese herbs,in treatment of aplastic anaemiaMETHODS:The animal model of mice with aplastic anaemia was established by 60Co irradiationThe therapeutic effects of BSSS I on aplastic anaemia were observed with the changes of peripheral blood cell count,bone marrow GM-CFU-c and activities of blood IL-1,IL-2 and IL-6RESULTS:After treatment with BSSS I,the peripheral blood cell count,amount of bone marrow karyocyte and GM-CFU-c of bone marrow of the aplastic anaemia mice elevated markedly,and the blood IL-1 and IL-6 activities increased obviouslyThe levels of IL-6 increased to the level of normal and showed positive relationship with IL-1(r=0904)The levels of IL-2 of the treatment group showed a decrease but with no significant differencesCONCLUSION:The results showed that BSSS I may improve the function of hematopoiesis by elevating the peripheral blood cells and the amount of bone marrow karyocytes,accelerating GM-CFU-c of bone marrow,and through moderating the activities of IL-1,IL-2,IL-6