Latest Implications of Next-Gen Sequencing in Diagnosis of Acute and Chronic Myeloid Leukemia

The spectacular progress which was present in the past few years in the field of genome sequencing, together with the appearance on the market of some high performance devices in this field, the reduction of the costs regarding the analysis of the samples and the standardization of some protocols, has led to the establishment and introduction of the new generation of sequencing techniques in clinical diagnostic labs. An important role is played by the implementation of this technique in the oncology clinics. In this context, we found it appropriate to discuss in this chapter about the role of next-gen sequencing in determining the genetic probabilities of occurrence of oncological pathologies in the healthy population, the screening of these diseases at the population level, the diagnosis and classification of this pathology, the establishment of the therapeutic conduct using the technique, as well as the progression of the disease. In this chapter, we intend to discuss in particular the involvement of this technology in hemato-oncological diseases

Granulocytic Sarcoma as the First Sign of Acute Leukemia in Childhood

Acute myeloid leukemia (AML) may rarely involve the orbit as a solid tumor termed granulocytic sarcoma. This report describes the case of a child who presented with rapidly progressive unilateral proptosis and was diagnosed as rhabdomyosarcoma. However subsequent examination of the peripheral blood film revealed AML. Thus proptosis may present as the initial manifestation of AML

Studies on the morphology of leukaemic blast cells in relation to haematological parameters

A combination of haematological parameters with morphological evaluation of peripheral blood and bone marrow blast cells is crucial for leukaemia diagnosis. FAB (French– American–British) classification is a simple and powerful diagnostic tool for leukaemia           in developing countries like India. Differentiation block in the early stages of haematopoiesis and morphological characteristics of leukemic blast cells are directly related to haematological parameters. The present study is an approach to increase understanding of the simple morphological FAB classification of leukaemia  in relation to haematological parameters. The present study revealed that Chronic Myeloid Leukaemia  (CML) was the most common type of leukaemia , followed by Acute Myeloid Leukaemia, Acute Lymphoid Leukaemia  (ALL), and Chronic Lymphoid Leukaemia  (CLL) in Nagpur. Most of the cases of Acute Leukaemia  had severe anaemia and thrombocytopenia. Highest variation was found in Total WBCs count of different types of leukaemia , particularly in different subtypes of AML. The present study also suggested that FAB classification is not outdated, but it does require continuous revalidation and other procedures for refinement.          &nbsp

Clinical and Hematological evaluation of leukemias, using cytochemical stains and immunophenotyping

INTRODUCTION: Leukemias are abnormal proliferation of hematopoietic cells, causing progressive infiltration of the marrow. It is the eleventh most common cancer in the world, and increasingly found now. Two widely used classification are used now, the FAB, and the WHO classification, which has got supplanted now, with increasing knowledge on cytomorphology and cytogenetics. This study attempts to evaluate the role of cytochemistry in the various types of leukemias, as well as the role of immunophenotyping in a select few cases, in patients which were referred to the Department of pathology, at Thanjavur Medical College. AIM OF THE STUDY: The main aim of the study was to assess the type and subtype of leukemia, using cytochemistry and their morphology, as well as to see the distribution of demographic data among them, and to evaluate their clinical and hematological presentation. RESULTS: 56 cases of leukemia, identified by morphology and cytochemistry, were seen in the study between January 2015 and June 2015. Cytochemical stains used were Sudan black B, and PAS. Out of the total, 36 cases were acute, rest were chronic cases. AML accounted for 43% of the cases, followed by CML at 33%, and ALL at 22%. A slight female preponderance was seen in the number of AML cases, but a male predominance in ALL. ALL was common in the child age group, followed by AML, and CML in the 21-40 age group, followed by AML.CLL was seen mostly above 40 yrs of age. Anemia was seen more in acute leukemias especially ALL, followed by AML. Total count values were seen high in CML, followed by AML. Platelet counts were less in acute leukemias, especially ALL, followed by AML. Splenomegaly was the commonest feature seen in 21 cases. Immunophenotyping was done in 6 cases, 4 cases were concordant, showing a 67% rate. CONCLUSION: In a setting where there is a lack of facilities for flow cytometry, as in majority of centres in the developing countries, morphology combined with cytochemistry, still serves as the best means in diagnosing leukemia cases

A study on the cytomorphologic structure of blood cells by vital staining II. Leukemic cells in the bone marrow

Leukemic cells were cytologically studied in the human bone marrow culture by the utilization of vital staining of Janus green B and neutral red. The minute cellular morphology of various types of leukemia was studied with special reference to their alterations in the course of the culture. The cytologic deviation of leukemic cells from the corresponding normal blood cells was clarified on monocytic leukemia, chronic myelogenous leukemia with the blastic crisis, chronic myelogenous leukemia, and acute and chronic lymphocytic leukemias.</p

A Study on the Occurrence and Subtyping of Acute Leukemias Based on Morphology and Cytochemistry.

Leukemia Is The Term Used For Haematopoietic Neoplasm, Presenting With Wide Spread Involvement Of Bone Marrow And Peripheral Blood By Blasts And Is The Most Common Cancerous Disorder In Children1. Virchow Recognized Leukemia As A Distinct Disease In 1845 When He Used The Term Weisses Blut To Describe The Findings In An Autopsy In Which The Ratio Of Red Corpuscles To “Colorless (In Mass White)” Was Reversed2. He Used The Term Leukemia For The First Time In 1847 And Subsequently, Described Lymphatic And Splenic Forms3. Friedreich Recognized Acute And Chronic Types In 1857. Neumann, In 1868, Identified The Bone Marrow As The Origin Of Leukemia And Used Myelogenous As A Provisional Term That Has Been Validated Over Time. The First Case Of Acute Myeloid Leukemia (AML) Was Originally Termed Acute Non Lymphocytic Leukemia In 1900. During The First Half Of The Century Most Of The Subcategories Of Myeloid Leukemia Were Identified By Light Microscopy With The Aid Of Cytohistochemical Stains And Were Described Based On The Resemblance To Normal Hematopoietic Precursors. The Recognition Of The Philadelphia Chromosome In Chronic Myeloid Leukemia (CML) By Norwell And Hungerford In 1960 Was The First Demonstration Of Recurring Chromosomal Abnormality In Leukemia. In 1976 FAB Classification Was Proposed, With Several Subsequent Modifications. The FAB Group Defines Seven Subsets Of AML. Four Based On The Percentage Of Maturing Cells And Three Based On Lineage.FAB Classifies Acute Lymphoblastic Leukemia (ALL) Into Three Types Based On Morphology As L1, L2, And L3. In The Latter Half Of The Last Century, Numerous Cytogenetic Abnormalities, Including Specific Translocations, Were Identified In Subtypes Of AML And Led To Changes In Classifications, As Proposed In 1997 By The WHO4. Cytogenetics Has Profound Effect On Prognosis And Treatment. The WHO Subdivides AML Into True Denovo AML Occurring Predominantly In Young To Middle-Aged Adults, With Recurring Cytogenetic Translocations Or Inversions And MDS Related AML Occurring In Elderly Adults Often With Complex Chromosomal Abnormalities The WHO Recognizes Therapy Related AML And Retains The Morphologic Subcategories Of The FAB In Cases Not Otherwise Classified. The Category Of Acute Leukemia Of Ambiguous Lineage Is Also Added. As Per WHO Classification (2001) The Alls Are Separated Under Three Broader Categories, Precursor T-Cell, Precursor B-Cell, And Mature B-Cell Neoplasm Based On Their Immunophenotype5. The High Cost Of Cytogenetics And Immunophenotypic Studies Stands In Way Of Routine Application In All Patients. Inspite Of Advances Made In Classification And Prognostic Factor Delineation, It Is Essential To Classify Leukemias Basically Into AML And ALL As They Differ Significantly In Management And Prognosis. This Can Very Well Be Done By Combination Of Morphology And Cytochemistry Which Has A Sensitivity Of More Than 92%6. In A Developing Country Like India All The Patients Do Not Have Accessibility To Cytogenetics And Immunophenotypic Studies. Hence Classification Was Hitherto Done By Morphology Alone In The Past Decades. This Situation Necessitated The Introduction Of Cytochemistry, Which Is An Improvement Over The Previous Method, Though Not Up To The Level Of Precision Obtained In Cytogenetic Studies. Hence The Present Study Has Been Taken Up To Subtype Leukemias Using Morphology And Cytochemistry

Extramedullary leukemia in children presenting with proptosis

<p>Abstract</p> <p>Background</p> <p>We highlight the orbital manifestations of acute myeloid leukemia and the role of peripheral blood smear in the diagnosis of these cases. A total of 12 patients who presented with proptosis and were subsequently diagnosed to have acute myeloid leukemia based on incision biopsy or peripheral blood smear were included in the study.</p> <p>Results</p> <p>A retrospective review of all cases of acute myeloid leukemia presenting to the Orbital clinic was performed. The age at presentation, gender, presenting features, duration of symptoms and fundus features were noted. In addition the temporal relationship of the orbital disease to the diagnosis of leukemia, laterality, location of the orbital mass, imaging features and the diagnostic tools used to diagnose leukemia were noted. The median age at presentation was 6 years. The male: female ratio was 0.7:1. None of these patients had been diagnosed earlier as having acute myeloid leukemia. The presenting features included proptosis in all patients, orbital mass in 5 (41.7%), visual symptoms in 2 (16.7%) and subconjunctival hemorrhage in one patient (8.3%). A diagnosis of acute myeloid leukemia was established by incision biopsy in 4 patients, subsequently confirmed by peripheral blood smear testing and bone marrow biopsy in 2 patients which revealed the presence of systemic involvement. Imprint smears of the biopsy identified blasts in 2 of 4 cases. In 8 patients presenting with ocular manifestations, diagnosis was established by peripheral blood smear examination alone which revealed a diagnosis of acute myeloid leukemia.</p> <p>Conclusion</p> <p>A peripheral blood smear should be performed in all cases of sudden onset proptosis or an orbital mass in children and young adults along with an orbital biopsy. It can always be complemented with a bone marrow biopsy especially in cases of aleukemic leukemia or when the blood smear is inconclusive.</p

A case of platelet refractoriness in a patient with acute myelogenous leukaemia and paraplegia: management in a low resource setting

Background: Acute myelogenous leukaemia is a type of acute leukaemia more commonly seen in adults than in children and usually presents with features of anaemia, neutropoenia and thrombocytopoenia (pancytopoenia). Red cell transfusions and the use of granulocyte colony stimulating factor usually correct the anaemia and neutropoenia respectively while platelet concentrates are required for correction of thrombocytopoenia. However, some patients develop platelet refractoriness where they fail to achieve expected increment in platelet count following platelet transfusions which may be fatal because severe thrombocytopoenia may lead to bleeding into vital organs including the brain. The aim of this report is to document the management of platelet refractoriness in a patient with acute myelogenous leukaemia and paraplegia, with correction of thrombocytopoenia in a low resource setting.Methods: Data was obtained from the case notes of a 14 year old male with acute myelogenous leukaemia and paraplegia who developed platelet refractoriness. A review of literature was done by searching on Google and PubMed.Results: A 14 year old male who presented with pancytopoenia and paraplegia was diagnosed with acute myelogenous leukaemia. He had multiple transfusions and developed platelet refractoriness. Despite severe thrombocytopoenia and platelet refractoriness, he was commenced on chemotherapy and achieved remission.Conclusion: Paraplegia is an uncommon presentation of central nervous system involvement in acute myelogenous leukaemia. Platelet refractoriness is a feared complication occurring in haematological malignancies. The acute myelogenous leukaemia complicated by platelet refractoriness was treated successfully in a low resource setting.Keywords: Platelet refractoriness; Acute Myelogenous Leukaemia; Acute myeloid leukaemia; paraplegia; AML; Nigeri