Distribution of common BCR-ABL fusion transcripts and their impact on treatment response in Imatinib treated CML patients: A study from India

Background: Philadelphia chromosome (Ph): Hallmark of CML is caused by reciprocal translocation between chromosomes 9 and 22 resulting in BCR-ABL fusion protein. Most commonly associated breakpoint with CML is M-bcr in exon 13 or exon 14, producing splice variant b2a2 or b3a2 respectively. The distribution of these transcripts and their influence on clinico-hematological parameters is variable. Impact of the fusion transcripts on treatment outcome in Imatinib treated CML patients is still a matter of debate. Aims/settings and design: We conducted this study on 400 CML-CP patients to look for the distribution of fusion transcripts i.e. b3a2 and b2a2, their clinico-hematological profile and impact on treatment response in patients treated with Imatinib. Material and Methods: CML-CP was diagnosed by reverse transcriptase PCR (RT-PCR) for the BCR-ABL fusion transcript. Real-time quantitative PCR (RQ-PCR) was performed on peripheral blood every 3-6 monthly to look for treatment response. Results: The overall frequency of b3a2 transcript was observed in 288 (72%) followed by b2a2 in 104 (26%) and hybrid fusion transcript (b3a2 + b2a2) was seen in 8 (2%) cases. MMR was attained in 198/288 (68.7%) patients with b3a2 transcript and 90/288 (31.3%) patients failed to achieve MMR after 12 months of Imatinib therapy. Among the patients with b2a2 transcript, 44/104 (42.3%) patients achieved MMR and 60/104 (57.7%) patients failed to achieve MMR after 12 months of Imatinib therapy. Conclusions: In conclusion, the frequency of b3a2 transcript was more as compared to b2a2 transcript. MMR was significantly higher in patients with b3a2 transcript as compared to patients with b2a2.

Association of JAK2V617F mutation with thrombosis in Indian patients with Philadelphia negative chronic myeloproliferative neoplasms

Background: It is still a matter of debate regarding the association of JAK2V617F mutation with thrombosis in BCR-ABL negative CMPN patients. The role of JAK2V617F mutation in increasing the thrombotic risk in CMPNs is yet unequivocal. Aims: To clarify the contribution of JAK2V617F mutation in thrombosis in CMPN patients. Settings and Design: This retrospective study was done to evaluate role of JAK2V617F mutation in thrombosis in CMPNs. Materials and Methods: 65 CMPN patients (PV, ET and PMF) were analyzed for JAK2V617F mutation using ARMS-PCR and detailed history of thrombosis was recorded in these patients. Statistical Analysis: P values were 2 tailed, and statistical significance was set at P < 0.05. Results: 46/65 were males and 19/65 were females [M: F: 2.4:1] with median age 46 years [range, 14-80 years]. Patients had median Hb 15.6 g/dl [range, 5.1-20.3], median TLC 10.7 × 109/l [range 2.4-216] and platelet count 360 × 109/l [range, 20-1859]. 32 were JAK2V617F positive and 33 were negative for this mutation. On comparing the prevalence of thrombosis in JAK2V617F positive patients with JAK2V617F negative patients, we observed that 20/32 (62.5%) JAK2V617F positive patients had thrombosis as compared to 16/33 (48%) in JAK2V617F negative patients (P = 0.04). We observed significant association of JAK2V617F mutation with thrombosis, however no association of this mutation with thrombosis was observed among the JAK2V617F negative patients. Conclusion: Our study suggests that JAK2V617F mutation may increase the risk of thrombosis in CMPNs. This finding could lead to risk stratification, setting up the treatment strategy in CMPNs.

Childhood T-lineage Acute Lymphoblastic Leukemia: Management and Outcome at a Tertiary Care Center in North India.

Objective: To assess the clinical features, prognostic factors and outcome of childhood T-ALL in comparison with B-lineage ALL, treated with a uniform treatment regimen (MCP 841). Setting: Pediatric oncology division of a tertiary care institution in Northern India. Design: Retrospective analysis of clinical data and survival outcome. Participants: 60 children with T-ALL and 139 with Blineage ALL, and less than 15 years of age treated over 15 years. Results: T-ALL was observed in 30%. High risk features at presentation (age 10 years, WBC >50,000/mm3, mediastinal mass, and CNS leukemia) were significantly more frequent in T-ALL as compared to B-lineage ALL (P=0.049, P<0.001, P<0.001 and P=0.02, respectively). Fifty five of 60 T-ALL patients (91.7%) achieved complete remission after induction therapy. There were 3 induction and 10 remission deaths while 11 (18.3%) relapsed. The overall survival and event-free survival of T-lineage ALL (61.5±7.6 and 49.9±7.4, respectively) were similar to that of B-lineage patients (68.7±4.7 and 47.1±5.1, respectively). National Cancer Institute risk groups emerged as significant prognostic factor for event free survival only in B-lineage patients. Conclusions: Even though high risk features were significantly more frequent in T-ALL, survival outcome was similar to that of B-lineage patients. None of the routinely described prognostic parameters significantly impacted survival.

Effect of zinc supplementation on cell-mediated immunity and lymphocyte subsets in preschool children.

OBJECTIVE: In a zinc supplementation trial (with a significant impact on diarrheal morbidity), to evaluate effect of zinc supplementation on cellular immune status before and after 120 days of supplementation. DESIGN: A double blind, randomized controlled trial with immune assessment at baseline and after 120 days on supplement. SETTING: Community based study in an urban slum population. SUBJECTS: Randomly selected children (zinc 38, control 48), had a Multitest CMI skin test at both times. In 66 children (zinc 22, control 34), proportions of CD3, CD4, CD8, CD16, CD20 cells and the CD/CD8 ratio were also estimated using a whole blood lysis method and flowcytometry. INTERVENTION: Zinc gluconate to provide elemental zinc 10 mg daily and 20 mg during diarrhea. MAIN OUTCOME RESULTS: Regarding CMI, the percentage of anergic or hypoergic children (using induration score) decreased from 67% to 47% in the zinc group, while in the control group it remained unchanged (73% vs 71%) (p = 0.05). The percentage of children deteriorating between first and second tests was significantly lower in the zinc group (13% vs 33%, p = 0.03). Regarding lymphocyte subsets, the zinc group had a significantly higher rise in the geometric means of CD3 (25%, p = 0.02), CD4 (64% p = 0.001), and CD4/CD8 ratio (73% p = 0.004) with no difference in CD8 and CD20. The rise in CD4 was significantly higher in the zinc as compared to the control group; the ratio of geometric means was 1.45 (95% CI, 1.03-2.01). CONCLUSION: Zinc supplementation improves cellular immune status, which may have been one of the mechanisms for observed impact of zinc supplementation on diarrheal morbidity.

Association between diarrheal duration and nutritional decline: implications for an empirically validated definition of persistent diarrhea.

In an empiric approach to develop the definition of persistent diarrhea, we evaluated the relationship between diarrheal duration and risk of ensuing clinically significant decline in nutritional status, in a cohort of 395 children < 24 mo. Weights were obtained at the onset of diarrhea (wt I) and after three months interval (wt II). The occurrence of an adverse outcome (AO) was defined as a decline of -- 5% in NCHS weight for age (% WFA) between weights I and II or death in this interval. The risk of AO was similar for episodes of / or > 7 days while it was substantially higher in episodes with > 14 days duration (45%) than for shorter duration episodes, relative risk (RR) = 2.5 (p < 0.001). Relative risk remained similar for duration thresholds of 21 (2.3) and 28 days (2.6). As episode durations greater than 14 days are associated with substantial elevation of the risk of clinically cogent sequelae, such episodes may be termed 'persistent' at least in terms of poor prognostic expectations.

Intestinal glucoamylase & other disaccharidases in children with protracted diarrhoea.

Brush border lactase, sucrase and glucoamylase activities were assessed in jejunal mucosal biopsy specimens from 34 children (median age 11 months; range 1.5-38) having protracted diarrhoea with failure to thrive and 8 well nourished children with normal jejunal mucosal histology (median age 10.2 months; range 2-37). All enzymes showed progressive decrease in activity which was directly in relation to increasing degree of mucosal injury (P less than 0.002). Lactase was significantly reduced even in patients with protracted diarrhoea and normal mucosa (P less than 0.05). Glucoamylase and sucrase were significantly reduced only in the presence of mucosal injury (P less than 0.01). Our data suggest that most children with protracted diarrhoea may not tolerate lactose containing feeds and may need lactose-free diets preferably based on starch. A small number of children with protracted diarrhoea, who have severe mucosal injury may not be able to handle even starch and may require diets based on short chain glucose polymers. The findings of this study, need to be corroborated with well-controlled metabolic balance studies.

Efficacy of nalidixic acid in the treatment of acute bloody diarrhea.

Of sixty four children (mean age 20.1 +/- 1.2 mo) with acute bloody diarrhea and high fever, 47 had infection with non-typhoidal Salmonella (NTS) (20), Shigella (15) and enteropathogenic E. coli (EPEC) (12) and were treated with nalidixic acid (NA). The mean duration (h) of presence of macroscopic blood in the stool following institution of treatment was significantly shorter (p less than 0.05) in those with EPEC (11.5 +/- 4.9) as compared to NTS (30.4 +/- 15.4) or Shigella groups (22.9 +/- 15.6). The number of children having less than or equal to 50% reduction in stool frequency within 72 h was: NTS (17); Shigella (14); EPEC (10). Negative stool cultures on day 5 were obtained in all patients with Shigella and EPEC and 16 (80%) of those with NTS. Two patients with NTS and one with EPEC failed to respond to NA.

Evaluation of a new latex agglutination kit for detection of human rotavirus in faecal specimens.

A commercial latex agglutination (LA) test was compared with ELISA and direct electron microscopy (EM) for detection of rotavirus antigen in 93 stool specimens obtained from as many children with acute gastroenteritis. Seventy one specimens (76.3%) were either positive or negative with all the three techniques, while 22 (23.7%) gave contradictory results. Only 1 sample was positive by LA test but not with ELISA or EM. The sensitivity of LA test and EM was 62.5 per cent (30 of 48) and 75 per cent (36 of 48); the corresponding specificity being 97.7 per cent (44 of 45) and 100 per cent (45 of 45) respectively. ELISA was more sensitive than the LA test and EM for detection of rotavirus antigen. LA test which is highly specific and a rapid method, may be useful in certain situations but its low sensitivity makes it unsuitable for use in routine clinical practice.

Aggregative Escherichia coli, Salmonella, and Shigella are associated with increasing duration of diarrhea.

In order to relate etiology of diarrhea to the duration of the episode, a cohort of 452 children up to 36 months of age was visited once weekly at their households for 18 consecutive months to record diarrheal morbidity. Fecal specimens were obtained in 453 diarrheal episodes occurring in 354 children during this period. The common putative agents as single isolations associated with diarrhea were EA-AggEC (17.2%), ETEC (14.1%), EPEC (6.0%) and rotavirus (4.0%). The pathogens with higher median duration of diarrhea were Shigella (13.5; mean +/- SD 13.7 +/- 2.7), Salmonella (8.8: 15.3 +/- 4.5) and EA = AggEC (12.0: 15.1 +/- 1.8). Of 55 episodes with duration of greater than 14 days, the main pathogens isolated were EA-AggEC (32.7%), ETEC (9.0), Salmonella, G. lamblia (5.5% each) and Shigella (3.5%). These data provide a preliminary evidence to suggest that EA = AggEC may well be the main long sought microbial agent responsible for diarrhea of long duration.