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Background: Carcinomas of the stomach are a heterogeneous group of lesions in terms of architecture, pattern of growth, cell differentiation, and histogenesis. Altered MUC5AC expression patterns have been reported previously in intestinal metaplasia as well as in gastric cancer. The aim of the study was to analyse the expression pattern of MUC5AC in normal, pre-neoplastic and neoplastic gastric epithelium.Methods: Formalin fixed paraffin embedded sections of sixty cases which include twenty cases of each normal gastric mucosa, intestinal metaplasia and gastric carcinoma were taken up for the study and subjected to immunohistochemistry using MUC5AC.Results: The intensity of MUC5AC immunostaining in normal gastric mucosa, intestinal metaplasia and gastric carcinoma was evaluated. Immunoreactivity was graded as 0 (negative), ± (trace positive), + (positive) or ++ (strongly positive). Statistical analysis was performed with Chi-Square test and significant differences were noted between these 3 groups (p value <0.05).Conclusions: Authors concluded that MUC5AC expression rates might be good parameters in progression of intestinal metaplasia to gastric carcinoma and might be a good prognostic marker for gastric carcinoma as it is very well implicated in understanding of gastric carcinogenesis.
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Background: To study the expression of PTEN (Phosphatase and Tensin homologue) and Cyclin D1 in normal, hyperplastic and neoplastic endometrium by immunohistochemistry and to corroborate the interrelationship between PTEN and Cyclin D1 in normal to neoplastic endometrial disorders including endometrial carcinoma.Methods: Formalin fixed paraffin embedded (FFPE) sections of spectrum of endometrium in fifty different cases were taken from secretory phase to endometrial carcinoma and subjected to Immunohistochemistry using PTEN and Cyclin D1 .Results: Immunoreactivity was regarded as positive when brown staining was localized in the nuclei or cytoplasm. The intensity of nuclear staining was graded from 0 to 3+ and the extent was semi quantitatively estimated. If less than 10% of cells were positive a score of 0 was given, 11 % to 30% cell positivity was scored as 1+, 31% to 60 % positivity was scored as 2+ and more than 60% positive cells was labelled as 3+. Statistical analysis was performed with Chi-Square test and significant differences were noted between these 3 groups (p value < 0.05).Conclusions: The present study supports that an inverse correlation exists in the expression of PTEN and Cyclin D1 in normal, hyperplastic and neoplastic endometrium. The decreased PTEN expression is a marker of the earliest endometrial premalignant lesions, and we propose that use of PTEN immunostaining may be informative in identifying premalignant lesions that are likely to progress to carcinoma. Cyclin D1 expression in endometrial glands increases progressively in intensity and extent from normal endometrium to hyperplasia to carcinoma.
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Background: Acute Myeloid Leukemia (AML) is a malignancy of the cells of myeloid series characterized by the rapid growth of Myeloblasts. The diagnosis of AML is established by demonstration of more than 20% of the blood and/or bone marrow by leukemic myeloblasts. Immunophenotyping is one of the most useful tool for the confirmation, lineage assignment and subtyping of leukemias. This study was aimed to phenotype and classify acute leukemias by flow cytometry using commonly used markers for leukemia diagnosis and to establish whether CD 117 can be considered as a lineage specific marker in diagnosis and subclassification of AML.Methods: Flow Cytometric Immunophenotyping was employed for the study. The myeloid antibodies employed in AML in our study included - CD117, CD11c, CD13, CD15, CD33, CD34, CD36, CD41, CD65 and MPO.Results: In our study AMLs constituted 46% of all acute leukemias. CD117 positivity was seen in 86.56% of the French American British (FAB) category of AML. The blasts gated using CD45 v/s SSC revealed variable expression of CD34, CD13 and CD33. The expression of CD117 was consistent particularly in AML-M0, AML-M1 and AML M2.Conclusions: CD117 is virtually a myeloid blast marker with a high sensitivity, specificity and positive predictive value. Among the various myeloid markers like cMPO, CD13, CD33 and CD117, it is just CD117 that has got a tremendous reproducibility in AMLs. Besides CD117 is a surface marker unlike MPO thus easier to process, time saving and less prone to nonspecific binding.
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We present a case of B-acute lymphoblastic leukemiain an elderly patient who presented with severe weakness and pancytopenia. The patient was a 75 year old Female whose blasts had an unusual morphology in form of coarse azurophilic granules and cytoplasmic blebs and on flow cytometry the blasts were present in the bright CD45 zone with a high side scatter. Bone marrow aspirate sample was subjected to multicolour flow cytometry using Beckman Coulter Navios® which is an 8 colour flow cytometer.Flow cytometricanalysis of the bone marrow aspirate showed blasts in the monocytic zone with a precursor B cell immunophenotype. Complete blood counts showed pancytopenia with peripheral blood film not showingany blasts. Bone marrow aspirate smears showed 20% blasts with coarse azurophilic granules and cytoplasmic blebs.The position of the blasts in this case which were in monocytic zone giving them a bright expression of CD45 and a high side scatter on the CD45 side scatter. This is not the usual positionfor blasts in B-acute lymphoblastic leukemia as these blasts are less complex. A bright expression of CD45 by blasts in B-acute lymphoblastic leukemia is known to be associated with a poor prognosis but the clinical significance of blasts being bright CD45 with a high side scatter is a very rare occurrence and more number of cases with a similar presentation are required to determine a prognostic significance.
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Background: This study was conducted to assess the CD34+ hematopoietic progenitor cells enumeration by flow cytometry and the utility of performing mononuclear cell count before performing the Stem cell enumeration. Collection was done on two consecutive days with CD34+ hematopoietic progenitor cell enumeration of both the samples. Mononuclear cell counts were done in all the patients. The purpose of the study was to do counts directly from the leukapheresis pack and see the reliability of this practice.Methods: Samples were collected from the leukapheresis pack and subjected to mononuclear cell count and CD34+ hematopoietic progenitor cells enumeration by flow cytometty before harvesting.Results: A total of 66 samples from 34 patients were taken up for the study. 76.47% of our cases were that of multiple myeloma and 17.64% of the cases were that of non Hodgkin lymphoma and 2.94% cases each of neuroblastoma and Hodgkin lymphoma. It was noted that the mononuclear cell counts correlated well with the CD34+ HPC in most of the cases with MNC being above 4 x 108 per pack per kg body weight in cases where CD34+ HPC counts were more than the desired lower limit of 2 x 106 per pack per kg body weight.Conclusions: It was observed that flow cytometric enumeration of CD34+ hematopoietic progenitor cells directly from the leukapheresis pack gave satisfactory results even without doing peripheral blood CD34+ HPCs enumeration before leukapheresis. Also, in our study we were able to set a limit of mononuclear Cell at 4 x 108 per pack/kg BW as counts beyond that always correlated with the more accurate flow cytometric method of CD34+ HPC count of more than 2 x106 per pack/kg body weight, therefore acting as a crude method for assessing the mobilization.
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Background: Breast cancer is the most frequent cancer in India. During the last few years, several investigators have focused on tumor angiogenesis as a critical step in cancer development and progression. Among these, vascular endothelial growth factor (VEGF) is emerging as a prognostic marker in patients with several type of cancer including breast cancer. The aim of the study was to analyse the expression of VEGF in human breast cancer as compared to normal breast tissue and benign breast lesions by immunohistochemistry. Also, to assess the usefulness of VEGF as a predictor of aggressiveness of breast lesions.Methods: Formalin fixed paraffin embedded sections of 10 cases of normal breast tissue, 20 cases of benign breast lesions and 20 cases of malignant breast lesions were taken up for the study and subjected to immunohistochemistry using VEGF.Results: The intensity of VEGF immunostaining in normal breast, benign and malignant breast lesions was evaluated and scoring was graded as 0, 1+, 2+, 3+ and 4+. Statistical analysis was performed with Chi-Square test and significant differences were noted between these 3 groups (p value <0.05).Conclusions:VEGF expression correlated well with the grade and stage of tumor indicating that VEGF positive tumors are biologically aggressive and are associated with poor prognosis but little is known about the implication of genetic alterations of VEGF in benign breast lesions.
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Introduction: The blasts in the peripheral blood is usuallyassociated with a haematological disorder. Study aimed tosee if a single blast seen in a peripheral blood film of healthyindividuals and patients of non-neoplastic haematologicaldisorders and non-hematological neoplasms/disorders didhave any clinical significance.Material and Methods: The period of study was from 2014to 2018 conducted at a tertiary care hospital. The study wasdone exclusively on Peripheral Blood Films. The films werestained with Leishman stain. An occasional blast was detectedin some healthy subjects and patients of non-neoplastichaematological disorders and non-hematological neoplasms/disorders.Result: The routine smears were analyzed with caution afterthe first case of a patient of Iron Deficiency Anemia revealeda clear-cut Blast. In the period of 4 years from 2014 to 2018,a total of 23 cases of apparently normal individuals or patientswith non neoplastic haematological disorders and nonhematological neoplasms/disorders showed at least 1% on oneseparate occasion.Conclusion: A careful morphological examination in theperipheral smears of few normal individuals and individualswith non neoplastic haematological disorders and nonhematological neoplasms/disorders showed at least 1%Blast on one separate occasion. The cells were clear cutblasts with a large size, fine nuclear chromatin, one to twonucleoli with round to slightly irregular nuclear contours anda mild amount of pale agranular cytoplasm. On thoroughinvestigations of these individuals there was no evidence ofa neoplastic haematological disorder. Therefore a single blastin a peripheral blood seen in a healthy individuals or patientswith benign haematological disorder and non-neoplasticneoplasms/disorders may not always indicate a neoplasticprocess.