Background & objectives: The usefulness of cation exchange high performance liquid chromatography (CE-HPLC) as a tool for detection of thalassaemia/haemoglobin variants was evaluated in a prospective study in a tertiary care centre in north India. We also tried to evaluate the effect of concurrent nutritional deficiency on the HPLC pattern in the local ethnic population. Methods: A total of 800 blood samples were analyzed on the Bio-Rad Variant HPLC system by β-thal short program. The retention times, proportion of the haemoglobin (%), and the peak characteristics for all haemoglobin fractions were recorded. Alkaline and acid haemoglobin electrophoresis was performed to document the identities of the haemoglobin variants, wherever necessary. Many cases were subjected to family studies for a definitive diagnosis. Results: Among 800 samples tested, 553 (69.1%) were found to have normal HPLC pattern. Apart from β- thalassaemia, nine additional variants were encountered; HbS (2.8%), HbE (2.5%) and HbD (1.1%) being the most common variants present. Other variants included Hb Q-India, Hb-Lepore, δβ-thalassemia/ HPFH, HbD-Iran, HbJ-Meerut and HbH disease. There was a significant decrease in the level of HbA₂ associated with iron deficiency anaemia (IDA) (P=0.004) and increase in megaloblastic anaemia (P<0.001) among subjects with normal HPLC pattern. Interpretation & conclusions: HPLC was found to be a simple, rapid and reliable method for the detection of hemoglobin variants. An accurate diagnosis can be provided in majority of cases by use of retention time, proportion of total haemoglobin, and peak characteristics of HPLC. Haemoglobin electrophoresis and family studies play a valuable role in difficult cases. Concurrent nutritional deficiency also has an effect on HbA₂ levels.

Background: Primary mediastinal lymphoma is an uncommon tumor. Hodgkin's lymphoma (HL), primary mediastinal B-cell lymphoma (PMBCL), and T-lymphoblastic lymphoma are the most common primary mediastinal lymphomas. Key morphological and immunohistochemistry (IHC) features play a very crucial role in diagnosis as well as further categorization. Materials and Methods: In this study, the morphological spectrum and histological features of 32 cases of primary mediastinal lymphomas diagnosed over 5 years were studied and morphological and IHC features of PMBCL versus HL were compared. Features of PMBCL were also compared against a control group of systemic diffuse large B-cell lymphoma. Results: Although PMBCL and HL are known to show overlapping morphological features, it was observed that presence of clear cells and compartmentalizing fibrosis in PMBCL; and classical Reed–Sternberg cells and dense inflammatory background in HL are important morphological clues while evaluating the biopsies. PMBCL showed diffuse, strong and uniform CD20 positivity; whereas CD30 showed focal/patchy, weak to moderate and heterogeneous expression, wherever found positive. As against this, HL showed diffuse, strong and uniform CD30 positivity; and focal/patchy, weak to moderate and heterogeneous CD20 expression, if found positive. CD20, CD3, and CD30 were sufficient in most of the cases while diagnosing PMBCL and HL. Conclusion: This study emphasizes the critical examination of IHC markers. Only positive expression in neoplastic cells is not sufficient to make a diagnosis, equal importance should be given to percentage, intensity, pattern, and type of positivity. Apart from basic IHC described above; CD15, leukocyte common antigen and fascin played an important role in differentiating HL and PMBCL in select doubtful cases.

Rrelate them with p53 expression. Materials and Methods: This was a retrospective study where 17 diagnosed cases of retinoblastoma were included. Cases were re-evaluated for various histomorphological parameters. Immuno-histochemical analysis was done with p53 antibody by Streptavidin biotin method. Results: The patients were in the age range of 1.5-50 years. Common histological features included necrosis (70.5%), calcification (64.7%), and retinal detachment (58.8%). Incidence of various morphological parameters was anterior chamber seeding (47.2%), ciliary body involvement (29.4%), iris involvement (29.4%), choroid involvement (58.8%), scleral invasion (29.4%), extrascleral invasion (11.8%), and optic nerve infiltration (23.5%). p53 expression was present in four cases out of 13 cases (30.7%) and showed a significant association with choroid invasion (P = 0.02). Discussion: The presence of HRF should alert the physician for a possible metastasis, and such patients should be kept on regular follow-up to detect an early recurrence. p53 expression, a known poor prognostic indicator, showed significant association with choroid invasion, however, no association was seen with other HRF. Conclusion: Histopathological HRF have significant therapeutic and prognostic implications. The incidence of HRF is higher in developing countries as patients present with a more advanced stage of disease. p53 expression is significantly associated with choroid invasion out of all HRF....