Absolute Lymphocyte Count at the End of Induction as a Surrogate Marker for Minimal Residual Disease in T-cell Acute Lymphoblastic Leukemia

Objective: The relation of absolute lymphocyte count (ALC) with minimal residual disease(MRD) in T cell – acute lymphoblastic leukemia (T-ALL) is not known. The objective of thestudy was to correlate ALC with MRD, steroid-response and complete remission (CR).Methods: De-novo T- ALL patients (age 1-18 y) recruited prospectively; 52 enrolled, 9excluded, and 43 analyzed. 39 achieved CR and MRD was available for 28 patients; 23 wereMRD negative. Results: ALC did not correlate with steroid response and CR. Median (range)ALC at the end of induction was significantly higher in patients who were MRD negativecompared to MRD positive [1.24 (0.12, 6.69) vs 0.62 (0.15, 0.87); P=0.03], respectively.Patients having ALC ≥700 ×109 /L were significantly more likely to be MRD negative thanthose with lower values (P= 0.028) Conclusion: Our study suggests that ALC is a favorablefactor, and may act as surrogate marker for MRD

Nucleophosmin mutation analysis in acute myeloid leukaemia: Immunohistochemistry as a surrogate for molecular techniques.

Background & objectives: Mutation of nucleophosmin (NPM1) gene in the absence of FLT3-ITD (FMS related tyrosine kinase 3 - internal tandem duplications) mutation carries a good prognosis in cytogenetically normal acute myeloid leukaemia (AML). NPM1, a multifunctional nucleolar phosphoprotein that shuttles between nucleus and cytoplasm, gets trapped in the cytoplasm when mutated. Immunohistochemical (IHC) demonstration of its aberrant cytoplasmic location (NPMc+) has been suggested as a simple substitute for the standard screening molecular method. This study was aimed to assess the diagnostic utility of IHC on formalin fixed bone marrow biopsies in comparison with the reference molecular method (allele specific oligonucleotide - polymerase chain reaction; ASO-PCR) to predict NPM1 mutation status in AML patients. Methods: NPM protein IHC was performed using mouse anti-NPM monoclonal antibody on 35 paraffin-embedded bone marrow biopsies of patients with primary AML of any French-American-British (FAB) subtype. Results of IHC were compared with those of ASO-PCR. Results: Of the 35 AML patients, 21 (60%) were positive for NPM1 exon 12 gene mutation by ASO-PCR, 19 (90.47%) of these 21 were NPMc+. Thirteen of the 35 patients were negative by both the methods. One NPMc+ patient was not detected by ASO-PCR. IHC had a sensitivity and specificity of 90 and 93 per cent, respectively, compared to the molecular screening gold standard. Interpretation & conclusions: Mutation of NPM1 determined by the widely available and inexpensive IHC agrees closely with results of the standard molecular methods. Thus, technically and financially not well endowed laboratories can provide the prognostically and potentially therapeutically important information on NPM1 mutation using IHC.

Alpha 1 Antitrypsin Deficiency in Children with Chronic Liver Disease in North India.

Objective: We attempted to determine the role of alpha-1- antitrypsin (AAT) deficient variants as an etiologic factor for chronic liver disease in North Indian children. Design: This study investigated 1700 children (682 retrospectively and 1018 prospectively) (840 CLD, 410 neonatal cholestasis and 450 without liver disease) for AAT deficiency. Setting: Tertiary referral center, All India Institute of Medical Sciences, New Delhi. Patients: Of 1250 liver disease patients, 98 (7.8%) were suspected to be AAT deficient on the basis of screening tests (low serum AAT levels and/or absent/faint alpha-1- globulin band on serum agarose electrophoresis and/or diastase resistant PAS positive granules on liver biopsy). Main outcome measures: AAT deficient Z or S allele in suspected patients. Results: Z or S allele was not observed on phenotyping (1700 subjects), or with PCR-RFLP, SSCP and sequencing done in 50 of 98 suspected AAT deficient patients. A novel mutation G-to-A at position 333 in exon V was found in two siblings having positive immunohistochemistry for AAT on liver biopsy, both of whom had significant liver disease with portal hypertension. Conclusion: In conclusion, AAT deficiency as an etiologic factor for chronic liver disease in childhood appeared to be uncommon in North India.

Multiple myeloma presenting with coexisting severe marrow hypoplasia.

A 68-year-old man was referred to us with clinical and bone marrow (BM) features compatible with aplastic anemia. The correct diagnosis, hypoplasia of the BM coexisting with multiple myeloma, became apparent after noting rouleaux in the peripheral blood (PB) and approximately 50% plasma cells in the touch imprint of one of the two BM biopsies done. As standard therapy was precluded, the patient was put on dexamethasone but died within 4 days. This first case of the coexistence of untreated myeloma with aplastic BM shows that even apparently straightforward hypoplasia seen on the BM biopsy should be interpreted in conjunction with the PB smear and the BM touch imprint findings. Among other things, the BM biopsy and imprint should be repeated if the PB has findings such as rouleaux that do not fit with straightforward aplastic anemia. The combination of myeloma and BM aplasia precludes standard therapy and is rapidly fatal.

Unusual lymphocytosis with systemic lupus erythematosus.

A 15-year-old boy with systemic lupus erythematosus, who on a follow up visit complained of recurrent episodes of fever, easy fatiguability, and seizures. Investigations revealed lymphocytosis (95%), anemia, and a positive PCR for cytomegalovirus (CMV). Electron microscopy of the lymphocytes revealed intranuclear inclusion bodies supporting the diagnosis of CMV infection. The child was treated with ganciclovir and discharged. At discharge the child was afebrile. However, lymphocytosis persisted even after 9 months of discharge. Repeated screening for possible lymphoreticular malignancy was negative. It is likely that lymphocytosis in this child was due to persistence of CMV infection in host cells leading to continued provocation of the host immune system.

Pelger-Huet anomaly: a case report.

Impaired lobulation of neutrophils together with exaggerated clumping of chromatin, characteristic of Pelger-Huet anomaly was seen as an incidentalfinding in a 43-year old man who presented with depression. Peripheral blood and bone marrow findings and cytochemistry of the abnormal cells are described and the disease entity discussed.

Melphalan and dexamethasone for patients with multiple myeloma who are not candidates for autologous stem cell transplantation.

BACKGROUND: Multiple myeloma is a disease for which a number of treatment options are available. The choice of therapy is often based on factors such as cost, ease of administration and faster response as the survival rates are similar with most of the regimens. We assessed the efficacy of a combination of melphalan and dexamethasone as first-line therapy in patients with multiple myeloma who were not candidates for autologous stem cell transplantation. METHODS: Thirty-four patients with multiple myeloma were included in the study. Patients received a maximum of 12 cycles of chemotherapy consisting of oral melphalan 8 mg/m2 on days 1-4 and oral dexamethasone 40 mg on days 1-4 and days 9-12 every 4 weeks. Patients were assessed for response on the basis of M proteins and a bone marrow biopsy with touch preparation. RESULTS: The median follow up of surviving patients was 40 months. Nine patients (26.1%) had complete response/near complete response (5 had negative immunofixation) and 15 (44%) had partial response. The regimen was well tolerated and there were no therapy-related deaths. The 3-year overall and progression-free survival rates using the Kaplan-Meier method were 53% and 34%, respectively. The median duration of overall and progression-free survivals were 58 and 28 months, respectively. CONCLUSION: The combination of melphalan and dexamethasone is safe and effective in patients with multiple myeloma who are not candidates for autologous stem cell transplantation.

Economical choices in setting up a diagnostic myeloma laboratory.

In setting up a diagnostic myeloma laboratory the popular, highly automated and otherwise excellent choices of equipment and laboratory practices, so exorbitantly raise costs that the sustainability, even in large government hospitals in third world countries may become difficult. Based on our experience in a regional cancer center in India, we offer here, guidelines for carrying out high resolution electrophoresis, densitometry, immunofixation and urine concentration. We show that by simply employing well established techniques and doing them properly, one can get results of excellent quality at minimum cost and minimum dependence on costly imports.

Etoposide, vinblastine, adriamycin and prednisolone (EVAP) combination chemotherapy as first-line treatment for Hodgkin's disease.

BACKGROUND: Mechlorethamine, vincristine, procarbazine, prednisolone (MOPP) and doxorubicin, bleomycin, vinblastine, dacarbazine (ABVD) are well established first-line chemotherapy protocols for the treatment of Hodgkin's disease. The aim of this study was to try a new combination of drugs that individually have a proven efficacy in Hodgkin's disease but have less incidence of severe nausea, vomiting, pulmonary toxicity and sterility. METHODS: This prospective, single-arm study enrolled 66 newly diagnosed, previously untreated patients of Hodgkin's disease with stages IA (bulky)-IVB disease. They were given 6-8 courses of etoposide, vinblastine, doxorubicin and prednisolone (EVAP) as first-line chemotherapy between January 1992 and December 1997. Radiotherapy (RT) was given to the involved fields of those patients who had bulky (> or = 10 cm) stages I or II disease at presentation. The end-points were (i) complete and overall response; (ii) disease-free and overall survival; and (iii) toxicity. RESULTS: Complete response was seen in 78.8% and partial response in 12.2% of patients; the overall response rate was 91%. The median follow up was 48 months. The 5-year overall and disease-free survivals were 72% and 62%, respectively. There were 3 episodes of grade IV neutropenia requiring hospitalization. One patient developed avascular necrosis of the femur. There were 2 deaths during treatment, one due to chemotoxicity, and another due to progressive disease. CONCLUSION: The overall and complete responses were fractionally inferior to the recently published hybrid MOPP/ ABV combination and that of ABVD chemotherapy. The advantages of the EVAP combination are absence of pulmonary toxicity, markedly lower incidence of sterility and nausea and vomiting. EVAP is an attractive option and a randomized trial is warranted to assess its efficacy against established protocols.