Clinical and hematological correlates of aberrant immunophenotypic profiles in adult and pediatric acute myeloid leukemia at presentation

Background: Aberrant phenotypes in acute leukemia have been reported with varying frequencies in independent studies and their association with prognostic factors is still a matter of debate. Aim: This study aims to identify the frequency of aberrant immunophenotypes in de novo acute myeloid leukemia (AML) and to evaluate their association with initial clinical and hematological features. Materials and Methods: A total of 181 patients of de novo AML were included during the time (July 2010–June 2012). The immunophenotype of all cases of AML was studied by using flow cytometry. Results: Aberrant lymphoid antigen expression was seen in 43.1% cases. Most frequent aberrant lymphoid antigen was CD7, seen in 26.5% cases. All French-American-British (FAB) subtypes except AML-M3 expressed aberrant lymphoid antigens. The expression was most common in AML-M4 in the current study. CD34 expression in AMLs was significantly associated with the expression of aberrant lymphoid antigens. Lymphoid antigen expression in adult AML was significantly associated with higher white blood cell (WBC) count (>50,000/mm3) and higher number of peripheral blasts (>70%). Conclusion: In summary, CD7 is the most common aberrant lymphoid antigen expressed in AML. FAB subtype AML-M3 is usually not associated with aberrant lymphoid antigen expression. AML cases with CD34 positivity are more likely to express aberrant lymphoid markers. The current study also supports that aberrant lymphoid antigen expression in adult AML is associated with adverse presenting hematological features (WBC count >50,000/mm3, peripheral blasts >70%). Pediatric Ly + AML cases are not associated with adverse presenting clinical and biological features.

Spectrum of AIDS defining & non-AIDS defining malignancies in north India.

Background & objectives: There is scarcity of data on the frequency of malignancies in HIV infected individuals from India. The objective of this study was to determine the type and frequency of malignancies in HIV infected individuals attending a tertiary care hospital in north India. Methods: The study design included retrospective analysis of data of all HIV infected individuals registered in the Immunodeficiency clinic from December 2009 to December 2011 and a prospective analysis of HIV infected individuals registered from January 2012 to April 2013. The clinical details and treatment outcomes of all individuals diagnosed to have AIDS defining and non-AIDS defining malignancies were recorded. Results: Records of 2880 HIV infected individuals were reviewed. Thirty one (19 males, 12 females) individuals were diagnosed to have malignancy. AIDS defining malignancy was found in the form of non-Hodgkin’s lymphoma in 12 individuals and cervical cancer in six women. Non-AIDS defining malignancies included Hodgkin’s lymphoma (n=2); and chronic myelogenous leukaemia, carcinoma base of tongue, carcinoma larynx, carcinoma bronchus, sinonasal carcinoma, ovarian carcinoma, anal carcinoma, carcinoma urinary bladder, pleomorphic sarcoma, parathyroid adenoma, and renal cell carcinoma in one individual each. Mean CD4+cell count prior to ART initiation was 250 ± 195.6 (median: 187; range, 22-805) cells/μl and at the time of diagnosis of malignancy was 272 ± 202 (median: 202; range, 15-959) cells/μl. The mean CD4+ count of individuals with AIDS defining malignancy was significantly lower when compared with non-AIDS defining malignancy (P<0.001). Fourteen individuals were alive and on regular follow up, 15 had died and two cases were lost to follow up. Interpretation & conclusions: The frequency of malignancies in HIV infected patients at our centre was 1 per cent, with non-Hodgkin’s lymphoma being the commonest. Further studies need to be done to document similar data from different parts of the country.

An Adolescent with Kawasaki Disease.

Kawasaki disease is an acute vasculitis of unknown etiology that predominantly affects children <5 years of age. The incidence and the severity of myocarditis in this disease is variable and depends upon the stage of the disease, acute or chronic. Acute-stage Kawasaki disease shows relatively high incidence of myocarditis, but almost all cases are clinically mild. We describe teenage boy presenting with atypical/incomplete manifestations of Kawasaki disease and developing fulminant myocarditis within a week of illness resulting in death. The case underscores the importance of suspecting Kawasaki disease in a young child presenting with features of myocardial ischemia.

Haematological profile of 21 patients with hairy cell leukaemia in a tertiary care centre of north India.

Background & objectives: Hairy cell leukaemia (HCL) is a B cell neoplasm which constitutes around 2 per cent of all the lymphoid leukaemias. It has a characteristic morphology and immunophenotypic profile. It is important to distinguish HCL from other B cell lymphoproliferative disorders due to availability of different chemotherapeutic agents. This study presents clinical, haematological and immunophenotypic profile of patients with HCL seen over a period of four years in a tertiary care hospital in north India. Methods: Twenty one cases of hairy cell leukaemia were analyzed for their clinical details, haemogram, bone marrow examination and immunophenotypic findings. Results: Age of the patients ranged from 28-76 yr with male predominance. Weakness and fever were commonest presentations. Splenomegaly, hepatomegaly, lymphadenopathy were seen in decreasing order of frequency. Anaemia was noted in all 21 patients, leukopenia in 15 and thrombocytopenia in 19 cases. Fourteen patients were pancytopenic. Bone marrow examination showed typical hairy cells in all cases. Immunophenotyping showed expression of CD19, CD20, CD103, CD25 and CD11c in all cases, while positivity was seen for CD79b in 93.7 per cent, kappa light chain restriction in 60 per cent and lambda in 40 per cent cases. Notably, 20 per cent showed CD10 and 12 per cent showed CD23 expression. Interpretation & conclusions: This study reveals some unusual findings in otherwise classical disease entity, like absence of palpable spleen, presence of lymphadenopathy, normal or elevated leukocyte counts, expression of CD10, which at times could be diagnostically challenging.

of multiplex polymerase chain reaction using IS6110 and Protein b for the diagnosis of extra-pulmonary tuberculosis: North India.

Background: Prompt and accurate diagnosis of extra-pulmonary tuberculosis (TB) is highly challenging. Current conventional techniques lack sensitivity and are time-consuming. Here, we report our experience with multiplex polymerase chain reaction (MPCR) using two targets namely IS6110 and protein antigen b in the diagnosis of extra-pulmonary TB. Materials and Methods: A total of 150 patients of extra-pulmonary TB visiting tertiary care center in north India between September 2008 and December 2009 were included in the study. Sixty-six biopsy samples and 84 were body fluids from these patients were subjected for microscopy (Ziehl-Neelsen), culture on LJ medium and for Multiplex PCR using IS6110 and Protein b antigen. Results: Smear positivity was noted in 11 samples (7.33%), and LJ culture yielded Mycobacterium tuberculosis in 8 biopsies and 9 body fluids with overall positivity of 11.3%. The multi-targeted PCR could detect M. tuberculosis in a total of 112 samples. Of 112 positive samples, only Protein b band was detected in 7 samples and only IS6110 was detected in 5 samples. Overall Protein b, PCR could detect 71.33% of the cases, whereas IS6110 was positive in 66.6% of the cases. Overall the sensitivities of microscopy, culture, IS6110 PCR, Protein b PCR and MPCR were 7.33%, 11.3%, 66.67%, 71.3% and 74.6%, respectively. Thus by using more than two targets the sensitivity increased from 66.67% of IS6110 to 74.6% in MPCR. Conclusion: Multiplex polymerase chain reaction using IS6110 and Protein b antigen is a highly sensitive and specific tool in the diagnosis of pauci-bacillary conditions like extra-pulmonary TB.

Viral markers in patients with hemophagocytosis: A prospective study in a tertiary care hospital.

Background : Hemophagocytic syndrome (HPS) is a rare clinicopathological condition characterized by the activation of macrophages with prominent hemophagocytosis in bone marrow and other reticulo-endothelial systems. HPS can be familial or secondary to infections including viruses. Aim : To study the viral markers in patients with HPS. Materials and Methods : Serum samples of patients with HPS and control group were screened for anti EBV VCA IgM, and IgG, anti-Parvo B19 IgM, and anti-CMV IgM antibodies using commercially available ELISA kits and CMV and ParvoB19 DNA by polymerase chain reaction (PCR). Results and Discussion : The present prospective study reports the profile of viral markers in HPS cases from north India. Among the 14 HPS cases 43% (6/14) were positive for at least one viral marker tested, of which EBV was found to be the most prevalent (3/6: 50%) followed by parvovirus B19(2/6: 33%) and cytomegalovirus (1/6: 17%). Mortality was noted in 33% of virus associated HPS patients. Our study highlights the higher association of Epstein-Barr virus (EBV) with HPS as compared to other viruses along with higher rate of mortality in both parvovirus B 19 and EBV associated HPS.

Philadelphia chromosome detection in chronic myeloid leukemia: Utility of phytohemagglutinin-stimulated peripheral blood culture.

Background: The conventional cytogenetic approach to demonstrate Philadelphia (Ph) chromosome at times does not yield enough number of metaphases or are of suboptimal quality. Further, the rapid molecular tests have completely pushed this simple technique into disrepute. Aims: This study aimed to evaluate usefulness of phytohemagglutinin (PHA)-stimulated peripheral blood culture for detection of Ph chromosome in chronic myeloid leukemia (CML) patients. Materials and Methods: Fifty-six patients, including 11 newly diagnosed cases of CML and 45 patients of CML on imatinib therapy showing the presence of Ph chromosome in unstimulated samples, were included in the study. Cytogenetic analysis was done on unstimulated samples, i.e. bone marrow aspirate, 24- and 48-h peripheral blood culture, and compared with PHA-stimulated 72-h peripheral blood culture. Results: The preparations from PHA-stimulated peripheral blood culture samples in all 56 patients yielded high number of good-quality metaphases. All the 11 (100%) newly diagnosed patients and 39/45 (87%) of the patients on imatinib therapy showed the presence of Ph chromosome in PHA-stimulated samples. Addition of PHA-stimulated 72-h peripheral blood culture preparation can be of use for increasing the diagnostic yield in cases of CML with suboptimal results on conventional cytogenetics from bone marrow aspirate sample.

Cytogenetic & molecular analyses in adult chronic myelogenous leukaemia patients in north India.

Background & objectives: Chronic myelogenous leukaemia (CML) is the commonest leukaemia in Asia. There is a paucity data on cytogenetic and molecular analyses of Indian CML patients. This apparently reflects the low availability of cytogenetic and molecular techniques in our country. This study aimed to document various types of BCR-ABL fusion transcripts in different phases of CML and to compare the Ph chromosome positivity/negativity vis-a-vis BCR-ABL fusion transcripts in adult CML patients. Methods: Between June 2004 and February 2009, 208 patients were diagnosed as CML in chronic phase (CP), accelerated phase (AP) and blast crisis (BC), according to standard criteria. Cytogenetic and molecular genetic analyses were performed in all patients. Various types of BCR-ABL hybrid transcripts were compared with phases of CML and cytogenetic abnormalities. Results: Among 208 CML patients, b3a2 BCR-ABL transcripts were most commonly detected (66.82%) followed by b2a2 (28.84%), b3a2 + b2a2 (3.36%), b3a2 + e19a2 (0.48%) and b2a2 + e19a2 (0.48%). b3a2 transcripts were more frequently detected than b2a2 transcripts, in the whole group of 208 as well as in 183 CML-CP patients (P<0.0001). Ph chromosome was positive in 135 of 139 patients with b3a2 transcripts and 56 of 60 patients with b2a2 transcripts, difference not being significant. Additional cytogenetic abnormalities detected in 3.8 per cent patients in CML-CP and 44 per cent patients in CMLAP/ BC, did not show predilection for any BCR-ABL transcript type. Interpretation & conclusions: This study documents higher Ph positivity (96.15%) by cytogenetic analysis among CML patients, as confirmed by qualitative reverse transcriptase-polymerase chain reaction (RT-PCR) analysis in a large patient group from north India. Both the techniques contribute towards understanding the disease biology, and have important implications for diagnosis and management of CML patients.

Dietary adequacy of HIV infected individuals in north India - A cross-sectional analysis.

Background & objectives: Dietary inadequacy is common in developing countries and so is in immunedeficient HIV infected individuals. Hence, an assessment of dietary patterns was done among a group of HIV infected individuals and compared with recommended dietary allowances. Methods: One hundred consecutive HIV infected individuals were interviewed from the Immunodeficiency Clinic of a tertiary care center at Chandigarh. Dietary intake was assessed by 24 h recall method. Mean carbohydrate, protein and fat intakes were evaluated. Mean difference in the calorie intake from recommended dietary intake was then calculated. Mean absolute CD4 cell count was calculated and correlated with BMI and mean calorie intake. Results: Mean weight and BMI of the individuals participated in the study was 58.6 ± 11.7 (range, 34 - 94) kg and 21.5 ± 3.7 (range, 13.6 - 36.7) kg/m2, respectively. Mean total calories intake was 1713 ± 292.8 (860 - 2525) calories/day and mean difference in the calories taken from the standard values was 249.5 ± 190.7 (10.6 - 967.5) calories/day. There was no significant correlation between CD4 cell count and total calories taken. Interpretation & conclusions: In HIV-infected individuals the energy intake was significantly lower than the recommended average intake. Hence, efforts should be taken to ensure that HIV-infected individuals have access to high-quality, nutritious food choices that promote optimal dietary patterns.

Imaging in adult patients with acute febrile encephalopathy: What is better computerized tomography or magnetic resonance imaging.

Context: Imaging techniques are commonly used by emergency physicians in a febrile comatose patient. Their utility requires judicious use of the available resources. In this study, we have compared the efficacy of cranial imaging techniques in adult patients with acute febrile encephalopathy. Materials and Methods: This prospective observational study enrolled 101 patients presenting to the emergency with fever of less than 15 days duration and altered sensorium. All the patients were subjected to routine investigations, detailed cerebrospinal fluid analysis, computerized tomograms (noncontrast followed by contrast enhanced), and magnetic resonance imaging of the brain. Final diagnosis was reached after considering the clinical, biochemical findings, imaging results, and response to therapy. The positive yield of radiological investigations was compared against the final diagnosis. Results: The patients were divided into three groups. Forty-eight had evidence of meningoencephalitis, 22 patients had pyogenic meningitis, and 20 were combined together in other group. In 12 patients, a definitive diagnosis could not be made. Only 37% patients were detected to have abnormal computerized tomograms and the most common abnormality was diffuse edema, which failed to point to an etiological diagnosis. Magnetic resonance imaging was abnormal in 62.75% cases and was able to suggest an etiological diagnosis in 100% cases of cerebral venous thrombosis, tubercular meningitis, 95% cases of meningoencephalitis, and 45% patients with meningitis. Conclusions: We can conclude that magnetic resonance imaging provides better information than computerized tomography in adult patients with acute febrile encephalopathy.

Abnormal chromatin clumping in leucocytes of Ph positive chronic myeloid leukemia cases - extending the morphological spectrum.

The syndrome of abnormal chromatin clumping is largely a morphological entity characterized by exaggerated chromatin clumping seen in the neutrophils. According to the recent World Health Organization (WHO) classification, it is categorized as a variant of atypical chronic myeloid leukemia (aCML) or Ph-negative CML. Most of the cases reported in literature have been negative for the Ph chromosome or the BCR-ABL gene. Till date, Ph positivity has been demonstrated in just one case. We report two more Ph-positive CML cases with abnormal chromatin clumping in neutrophils. To the best of our knowledge, this is only the second time in literature that such cases have been described. These two unusual cases go on to extend the morphological spectrum of granulocytic changes seen in Ph-positive CML.

Proliferative indices, cytogenetics, immunophenotye and other prognostic parameters in myelodysplastic syndromes.

Thirty-five adult myelodysplastic syndrome (MDS) patients were included in this study: 11 refractory anemia (RA), 4 RA with ring sideroblasts (RARS), 9 RA with excess of blasts (RAEB), 10 RAEB in transformation (RAEB-T) and 1 chronic myelomonocytic leukemia (CMML). The ranges of survival were 4-51 months, 40-59 months, 7-38 months, 5-24 months and 5 days, respectively. Three patients died and 3 showed disease progression during the course of the study. A composite analysis of proliferative indices, cytogenetics, immunophenotype and other conventional/novel prognostic parameters in the context of Indian MDS patients was performed. The proliferative indices (AgNOR and Ki 67 positivity), immunophenotypic markers, serum LDH and ferritin levels revealed wide variations and great overlap among different FAB subtypes. The scoring systems (Bournemouth, Dusseldorf and Goasguen) did not correlate with the prognosis and survival (p > 0.05). Clonal cytogenetic abnormalities were detected in 24/35 (68.57%) patients, +8, -5 and -7 being observed commonly. Cytogenetic abnormalities were more frequent in RAEB (88.8%), RAEB-T (80.0%) and RA (63.6%) subtypes of MDS. By Using Mufti's prognostic system and International prognostic scoring system (IPSS), a good positive correlation was found between low risk category and RARS with better survival as compared to other risk categories/FAB subtypes (p < 0.01). However, rest of the FAB subtypes were assigned into high, intermediate and low risk categories without any correlation with the survival and/or leukemic transformation. RARS subtype revealed itself as the better prognostic category according to the cytogenetic findings as well as Mufti's grading system. This was possible due to the longer follow up available for these patients (40-59 months).

Improvement in CD 4 counts in HIV positive patients treated with HAART and antitubercular drugs: an observational study from North India.

A decrease in CD4 counts in HIV positive patients with concomitant tuberculosis leads to an increase in the morbidity and mortality. Little data exists about the use of antiretroviral drugs along with antitubercular drugs on the improvement in CD4 counts from this part of country. The records of 119 HIV and TB positive patients were obtained from immunodeficiency clinic of tertiary care centre of North India who were on drug treatment for both the diseases and were analysed for demographic profile and effects on CD4 counts. There was a statistically significant improvement in the CD4 counts of the patients as compared to their baseline values mean (SD) as 120.03 (124.1) at visit one to 270.2 (141.3) at visit two (p < 0.01) and 320.9 (184.3) at visit three (p < 0.05). Six patients died during the period of evaluation. Concomitant use of antitubercular drugs with antiretroviral drugs has resulted in a significant improvement in the CD4 counts which is a marker of delay in disease progression.

Acute myeloid leukemia (AML-M2) with mast cell hyperplasia of bone marrow: a report of three cases.

Bone marrow mastocytosis, though infrequently documented in Indian patients, may be observed in association with many non mast cell hematological neoplasms, including acute myeloblastic leukemia (AML) and myelodysplastic syndromes (MDS). We report three cases of acute myeloid leukemia with excess of mast cells in the bone marrow (BM) samples. Mast cell hyperplasia may remain under diagnosed due to shortcoming of morphological identification and diagnostic workup.

Multiple constitutional aetiological factors in bone marrow failure syndrome (BMFS) patients from north India.

BACKGROUND AND OBJECTIVES: A large number of patients diagnosed with bone marrow failure syndromes (BMFS), comprising aplastic anaemia (AA) and myelodysplastic syndromes (MDS), remain aetiologically uncharacterized worldover, especially in resource constrained set up. We carried out this study to identify a few constitutional causes in BMFS patients attending a tertiary care hospital in north India. METHODS: Peripheral blood lymphocyte cultures were performed (with and without clastogens) in a cohort of 135 consecutive BMFS patients, in order to detect Fanconi anaemia (FA), Down's syndrome (+21), trisomy 8 (+8) and monosomy 7 (-7). RESULTS: Constitutional factors were detected in 17 (12.6%) patients. FA defect was observed in 24.07 percent (13/54), 16.66 percent (1/6) and 2.85 percent (1/35) paediatric aplastic anaemia, paediatric MDS and adult MDS patients respectively. Down's syndrome was detected in 5.00 percent (2/40) adult aplastic anaemia patients. None of the patients revealed trisomy 8 or monosomy 7. INTERPRETATION AND CONCLUSION: Presence of an underlying factor determines appropriate management, prognostication, family screening and genetic counselling of BMFS patients. Special tests required to confirm or exclude constitutional aetiological factors are not available to majority of the patients in our country. Diepoxybutane (DEB) test yielded better results than mitomycin C (MMC) test in our experience.