Investigating the Association between the Autophagy Markers LC3B, SQSTM1/p62, and DRAM and Autophagy-Related Genes in Glioma.

High-grade gliomas are extremely fatal tumors, marked by severe hypoxia and therapeutic resistance. Autophagy is a cellular degradative process that can be activated by hypoxia, ultimately resulting in tumor advancement and chemo-resistance. Our study aimed to examine the link between autophagy markers' expression in low-grade gliomas (LGGs) and high-grade gliomas (HGGs). In 39 glioma cases, we assessed the protein expression of autophagy markers LC3B, SQSTM1/p62, and DRAM by immunohistochemistry (IHC) and the mRNA expression of the autophagy genes PTEN, PI3K, AKT, mTOR, ULK1, ULK2, UVRAG, Beclin 1, and VPS34 using RT-qPCR. LC3B, SQSTM1/p62, and DRAM expression were positive in 64.1%, 51.3%, and 28.2% of glioma cases, respectively. The expression of LC3B and SQSTM1/p62 was notably higher in HGGs compared to LGGs. VPS34 exhibited a significant differential expression, displaying increased fold change in HGGs compared to LGGs. Additionally, it exhibited robust positive associations with Beclin1 (rs = 0.768), UVRAG (rs = 0.802), and ULK2 (rs = 0.786) in HGGs. This underscores a potential association between autophagy and the progression of gliomas. We provide preliminary data for the functional analysis of autophagy using a cell culture model and to identify potential targets for therapeutic interventions.

Immunohistochemical Profile of Hodgkin and Non-Hodgkin Lymphoma.

OBJECTIVE: To analyze the frequencies of histological types of lymphoma, diagnosed with complete immunohistochemical profile in younger and older age group. STUDY DESIGN: Cross-sectional analytical study. PLACE AND DURATION OF STUDY: Dow Diagnostic Research and Reference Laboratory, Dow University of Health Sciences, Karachi, from January 2009 to September 2013. METHODOLOGY: Consecutive cases of lymphomas, which were diagnosed using immunohistochemistry, were analyzed according to WHO classification. Frequency and percentages for different types of lymphomas were calculated. Hodgkin and non-Hodgkin lymphomas characteristics in two age groups of less than and more than 40 years were compared, applying chi-square test. RESULTS: Out of the 318 cases, 79 (25%) were Hodgkin Lymphomas (HL) and 239 (75%) were Non-Hodgkin Lymphomas (NHL). Mixed Cellularity Hodgkin Lymphoma (MCHL) was the commonest (n=48). Amongst the NHL, 215 (89.95%) were B cell lymphomas and 24 (10.05%) were T-cell lymphomas. Diffuse Large B-Cell Lymphoma (DLBCL) was the commonest lymphoma (n=165, 69.95% of NHL). Anaplastic T-Cell Lymphoma (ALCL, n=10) was the commonest T-cell lymphoma. The frequency of HLwas significantly higher in the younger age group and that of NHLwas higher in the older age group (p < 0.001). Primary lymph node involvement was reported in 175 (55%) and cervical lymph node was the most frequent site. Extra nodal involvement was seen in 93 (29%) of all cases and was reported in 87 (36.4%) of NHLand 6 (7.5%) of HL. The most common extra nodal site was the gastrointestinal tract. CONCLUSION: Hodgkin lymphoma comprises 25% and non-Hodgkin lymphoma comprises 75% of all lymphomas. Both occur in younger age groups than reported in the West. B-cell NHLis three times more common than T-cell lymphoma. DLBCLis the most frequent lymphoma. ALCLis the most common T-cell, and mixed cellularity is the most common Hodgkin lymphoma.

Frequency and clinicopathologic correlation of different types of non Hodgkin's lymphoma according to WHO classification.

OBJECTIVES: To analyze the frequency and clinicopathologic correlation of different types of Non Hodgkin's Lymphomas (NHL) according to WHO classification of lymphoid neoplasms. METHODS: Total sixty two consecutive biopsy proven cases of NHL, from October 2008 to June 2010, were selected. The inclusion criteria was, all newly diagnosed patients of NHL with appropriate clinical information regarding age, gender, anatomic location and occurrence of B symptoms. All the cases were evaluated on Haematoxylin and Eosin (H & E) and special stains. Cases were subjected to Immunohistochemistry (IHC) using extensive panel of antibodies and classified according to WHO classification of lymphoid neoplasms. RESULTS: Clinical data showed that 42 (67%) were males and 20 (33%) females. The male to female ratio was 2.6:1. The age range was 6 to 80 years. Mean age for males was 39.6 +/- 17.3 years and for females 45.1 +/- 17.8 years. The B cell lymphoma comprised of 85.5% as compared to T cell lymphoma consisting of 14.5%. The extra nodal involvement was seen in 25 (40.3 %) cases, while 37 (59.3%) cases showed nodal involvement. The B symptoms were found in overall 29 (46.7 %) cases. CONCLUSION: B cell NHL is more common as compared to T cell lymphoma. Diffuse large B cell lymphoma (DLBCL) was the most frequent B cell lymphoma. The major bulk of T cell lymphomas comprised of anaplastic large cell lymphoma (ALCL). Significant association was seen in the occurrence of B symptoms with extra nodal origin and male gender.

Frequency of Helicobacter pylori in biopsy proven gastritis and its association with lymphoid follicle formation.

OBJECTIVE: To determine the frequency of H. pylori infection in biopsy proven gastritis and its association with lymphoid follicle formation. METHODS: This was a cross sectional study conducted at the Department of Histopathology, Dow Diagnostic Reference and Research Laboratory between January 2008 and December 2009. Analysis of 185 gastric antral biopsy specimens was done. Tissue sections were stained with haematoxylin and eosin for histological examination for severity of gastritis and lymphoid follicle formation. Giemsa stain was used for H. pylori assessment. RESULTS: Out of 185 cases, H pylori was found in 114 (61.6%) patients. Frequency of H. pylori infection was seen in fourth and fifth decades as 17.8% and 15.1% respectively. A total of 51 (27.6%) cases in which lymphoid follicle formation was found, 44 (38.6%) were associated with H. pylori infection. This association was statistically significant (p value < 0.0005) by using Chi square test. CONCLUSION: The frequency of H. pylori infection is common in our population, moreover, significant association is seen between lymphoid follicle formation and H. pylori infection.

Correlation of Ki 67 proliferative index with clinical and pathological features on tissue sections of non Hodgkins lymphoma by immunostaining.

OBJECTIVES: To assess Ki 67 proliferative index (PI) in tissue sections, with relation to grade and clinical parameters associated with non Hodgkin's lymphoma in Pakistani population. METHODS: All cases of non Hodgkin's lymphoma, diagnosed at histopathology section of Dow Diagnostic research and reference laboratory from October 2008 to June 2010, were included for this cross sectional study. Immunohistochemical study using monoclonal antibody MIB 1, Dako, Denmark against Ki 67 antigen was carried out on paraffin embedded tissue blocks of 62 patients. Ki 67 PI, with cutoff 45% was assessed in relation to specific phenotype, age, gender, site of origin and B symptoms. Mean Ki 67 PI was also calculated considering specific phenotype according to WHO classification. RESULTS: Out of 62 patients, Ki 67 PI > 45% was noted in 39 (62.9%) cases, whereas, in 23 (37%) cases the PI was < 45%. The mean Ki 67 PI was highest in Burkitt's lymphoma. Furthermore Ki 67 PI > 45% was seen in 21/25 (84%) cases with extra nodal involvement and 24/29 (82.7%) cases with B symptoms. CONCLUSION: Mean Ki 67 PI can discriminate the indolent versus aggressive behaviour of disease. In the study population significant association of high Ki 67 PI > 45% was found in relation to B symptoms and site of involvement, in terms of extra nodal origin, for non Hodgkin's lymphoma. These correlations demonstrate the significant role of high Ki 67 PI, to establish the proliferative activity of tumour as prognostic index marker along with clinical parameters at the time of diagnosis.

Clinicopathologic evaluation of subgroups of diffuse large B cell lymphoma by immunohistochemistry.

UNLABELLED: Diffuse large B cell lymphoma (DLBCL) has become an emerging epidemic in recent years. Striking heterogeneity in its clinical, biological and treatment responses prompted us to identify variation in our study group. The aim was to classify the DLBCL into prognosis-based subgroups according to the WHO classification and to evaluate their relation to clinical parameters (age, gender, anatomic location and B symptoms), as well as bcl 2 and Ki 67 status. PATIENTS AND METHODS: A cross sectional study was carried out on 42 DLBCL patients, classified histologically and immunophenotypically into germinal center B cell like (GCB) or non-GCB type. Immunohistochemistry (IHC) was performed using antibodies against CD 10, MUM-1 and bcl 6; additionally anti-apoptotic protein bcl 2 and proliferative marker Ki 67 (using cutoff value of 70%) were also assayed by IHC. RESULTS: Of the total 27/42 (64%) were males and 15/42 (36%) females, with a mean age of 44.1∓15 years. 15/42 (36%) cases were of GCB type as compared to 27/42 (64%) of non GCB type. Extranodal involvement and B symptoms were seen in 18/27 (66.6%) and 20/27(74%) of the non GCB type, whereas bcl 2 protein expression and Ki 67 proliferative index (PI) <70% were each noted in 22/27 (81.4%). CONCLUSION: We document an astonishingly high number of non-GCB type DLBCL in our population. It is alarming to see such an aggressive tumor proliferating in our region. Significant association of non-GCB type with extranodal origin, B symptoms and low Ki 67 PI (<70%) is another concern.