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Introduction: Excision of intra-axial tumours with maximal resection and minimal deficits is one of the intricate procedures in neurosurgery. Fluorescence-guided Surgery (FGS) helps in better identification of tumour tissue and allows Maximal Safe Resection (MSR) of high-grade brain tumours. Sodium fluorescein dye, when injected intravenously, gets concentrated in High-grade Glioma (HGG) tissue due to a disrupted Blood-Brain Barrier (BBB), thus aiding in MSR, Extent Of Resection (EOR), and Gross Total Resection (GTR). Aim: To assess the effectiveness of intraoperative use of sodium fluorescein in achieving MSR and GTR. Materials and Methods: This was a cross-sectional study conducted on 30 patients with HGG who underwent surgery between January 2021 and May 2023 at SDM College of Medical Sciences, Dharwad, Karnataka, India. Two groups of 15 patients each were divided into those operated with the use of Fluorescein Sodium (FS) and those without FS. FS was administered at a dose of 5 mg/kg at the time of craniotomy. The primary endpoint was the number of patients with Histopathological Examination (HPE)-confirmed HGGs showing no contrast-enhancing tumour in the immediate postoperative Magnetic Resonance Imaging (MRI). Secondary endpoints included residual tumours on postoperative MRI, Focal Neurological Deficits (FND), and FS-related toxicity. The association between FS status and recurrence at different treatment time points was assessed using the Pearson Chisquare test. Recurrence rates were compared in each FS group using the Cochran Q test. Statistical significance was set at a 5% level (p-value<0.05). Results: Homogeneous or heterogeneous yellow-green fluorescence was observed on the tumour tissue in all 15 patients (100%). Of the 15 patients where FS was used, two showed lesion recurrences at six months, and 13 showed recurrences at 12 months. Among the 15 patients where FS was not used, 11 showed recurrences at six months, and four showed recurrences at 12 months. This indicates that MSR, GTR, and EOR can be improved using FS intraoperatively during tumour resection. Conclusion: Based on the results, fluorescein dye with a 560 nm yellow filter under a highly illuminated microscope can help achieve better tumour resection with minimal injury to surrounding eloquent areas. It can be utilised as an ideal adjunct for tumour resection rather than with a microscope alone. [ABSTRACT FROM AUTHOR]

Introduction Glioblastoma multiforme (GBM) is a devastating form of cancer with a poor prognosis despite available treatments. Managing recurrent GBM remains challenging and lacks guidelines. This study aims to provide practice patterns for managing recurrent GBMs in India. Methods A panel of experts was assembled to develop practice patterns using the Delphi technique. Their responses were analyzed anonymously to ensure impartiality and generate recommendations. The statements were intended to be nonbinding and focused on promoting best practices in the field, without legal or regulatory authority. Results A total of 23 experts participated in the study, providing their opinions on various aspects of managing recurrent GBM. Consensus was achieved on individualized and multidisciplinary management as the preferred approach. Surgery in combination with other treatments was found to impact survival in patients older than 65 years, with re-surgery and adjuvant radiation and chemotherapy being the preferred options. Gadolinium-enhanced magnetic resonance imaging (MRI) brain with spectroscopy and diffusion-weighted imaging was favored. Molecular profiling was considered significant, with O 6 -methylguanine DNA methyltransferase methylation being most relevant. Surgery was recommended for recurrent GBMs, primarily based on Karnofsky's performance score (KPS). Surgical adjuncts such as neuronavigation and intraoperative MRI were considered valuable. Radiation therapy, specifically stereotactic radiosurgery, was recommended for selected cases, while opinions on re-chemotherapy were divided. Palliative care was deemed important. Conclusion This study presents practice patterns for managing recurrent GBM in India, providing standardized recommendations for practice. By implementing these, clinicians can make informed decisions, leading to improved patient outcomes and reduced variability in the management of recurrent GBM. [ABSTRACT FROM AUTHOR]

Brain tumors have always fascinated and intrigued histopathologists due to their diverse morphology and rarity. Recent surge in the molecular developments has further posed a challenge in diagnosis especially in a resource limited setting. Therefore, comprehensive tumor registries have become quintessential to compare our existing database with new found knowledge. A descriptive retrospective study was carried out on archival data of 5 years in a neuroscience institute. All neurosurgical cases operated with complete clinical history and definitive histopathological diagnosis were included. The cases were analyzed with reference to age, sex, location of the lesion, grade of the tumor, and immunohistochemical profile as available and compared with existing registries and literature. The primary brain tumors accounted for 38.29% of all pathologies. Majority of cases clustered around 40–70 years of age (65%). Pediatric (0–19 years) cases comprised 7%. Most common primary brain tumor found in the adult population was meningioma (28%) followed by Glioblastoma (25%). The most common neoplasm in pediatric age group were gliomas (46.29%) followed by embryonal neoplasms. Pituitary adenomas (PAs) constituted 16% of all intracranial neoplasm. Gonadotroph adenoma was the most common of the nonfunctional adenomas constituting one half of the PAs (51.72%). Somatotroph adenoma was most common in the functional group constituting 20% of all PAs. The layout of cases when compared with available brain tumor registries showed nearly similar trends in distribution. Our study succored data from the population in eastern part of India of which our institute is a major referral centre for neurosurgical cases. [ABSTRACT FROM AUTHOR]

Surgery of eloquent area gliomas is challenging and requires monitoring of the nearby white fiber tracts. In the present study, we analyzed 102 patients with eloquent region gliomas and discussed the concept of intraoperative dynamic white fiber tract navigation and monitoring. A total of 102 patients with an eloquent area glioma (52 insular, 29 motor area, 21 temporoparietal) were evaluated. The position of the white fiber tracts (corticospinal tract [or motor fiber; CST], inferior fronto-occipital fasciculus [ventral language fiber; IFOF], superior longitudinal fasciculus [SLF], and arcuate fasciculus [dorsal language fiber; AF) was recorded. Awake mapping of the cortical and subcortical eloquent structures was performed for all 102 patients. The suction stimulator was coregistered and used as a dynamic stimulator navigator. Of the 102 patients, 60 were men and 42 were women, with an average age of 39.8 years. Most of the white fiber tracts were normal (CST, 31.3%; IFOF, 39.2%; SLF/AF, 40.19%) or displaced (CST, 59.8%; IFOF, 47.05%; AF/SLF, 44.11%). A few were disrupted (CST, 8.8%; IFOF, 13.7%; SLF/AF, 15.7%). The extent of tumor resection was 82.8%, 86.5%, and 94% for those with insular glioma, motor area glioma, and temporoparietal glioma, respectively. Of the 102 patients, 18 had developed transient speech and language disturbances with improvement, and 14 had developed motor deficits, of whom, all except for 2, had shown gradual improvement. When the dynamic suction stimulator navigator was used, the extent of resection was 96.5%, without any added deficits. The use of intraoperative neuronavigation and neurophysiological assessment can help achieve maximal tumor resection of eloquent area gliomas. Use of the integrated suction stimulator navigator provided dynamic navigation and mapping of the peritumoral eloquent fibers. [ABSTRACT FROM AUTHOR]

Introduction: Isocitrate Dehydrogenase (IDH) plays an important role in cellular metabolism. In gliomas, the mutational status of IDH1/2 genes have paramount significance, however, study from Western India is limited. Therefore, the current study we sought to explore the clinical impact of IDH1/2 mutations for glioma patients from Western India. Materials and Method: A total of 50 pre-therapeutic, histopathologically confirmed patients with astrocytoma tumors were included and IDH1/2 mutations were detected using real-time PCR. IDH1/2 mutations were correlated with clinicopathological parameters and disease outcome. Data was evaluated by SPSS software. Results: The overall incidence of IDH1/2 mutations was noted in 24% (12/50) of glioma patients. Out of 12 patients whose tumors showed IDH mutations, 83% patients have IDH1 mutations, whereas 17% showed IDH2 mutation. Further, in IDH1 mutations, IDH1 R132H & IDH1 R132C mutations were noted in, 80% and 20% of patients, respectively. When correlated with clinicopathological parameters, significant inverse correlation was found with patients age (χ²= 9.75, r = -0.476, p=0.001) and grade of tumors (χ²=17.51, r =-0.636, p=0.0001). In Kaplan-Meier survival analysis, a part from age (Log rank=5.443, p=0.020), IDH mutation status (Log rank=3.855, p=0.050), and both, IDH mutation and low grade glioma tumors (Log rank=6.492, p=0.039) remained significant parameters for predicting better 24 months PFS and OS of glioma patients. However, in multivariate survival analysis using Cox Proportional Hazard Forward Stepwise Model, only combination of low grade glioma with presence of IDH mutation emerged at step one as positive significant independent prognostic factor that predict better PFS (HR=2.92, 95% CI=1.12-7.61, p=0.028) and OS (HR=3.0, 95% CI=1.45-6.19, p=0.003). Conclusion: Based on this data, we concluded that for glioma patients, apart from patients age, low grade tumors with presence of IDH mutations remained significant independent positive prognosticators and would be helpful to clinicians for better management of glioma patients. [ABSTRACT FROM AUTHOR]

Introduction: Advanced radiological imaging technology allows detailed anatomical view for diagnosis of diseases. The prevalence of incidental Magnetic Resonance Imaging (MRI) findings is relatively higher. Aim: To identify and describe the Incidental Findings (IF) in patients undergoing MRI brain and spine at a tertiary care centre in Southern Rajasthan, India. Materials and Methods: The cross-sectional observational study was carried out for a period of 18 months on patients referred to the Department of Radiodiagnosis for MRI brain and spine at Geetanjali Medical College and Hospital, Udaipur, Rajasthan, India, where report showed any incidental neuroradiological abnormality were included in the study. Patients were subjected to MRI machine (1.5 tesla SEIMENS-MAGNETOM Avanto OR 3 tesla GE SIGNA Architect) with T1-T2 weighted sequences. Incidental MRI findings with provisional diagnosis and clinical history were recorded and analysed for prevalence and proportion of IF (along with age and anatomical location specific IF). After entering data into Microsoft Excel 365, it was analysed for prevalence and proportion of IF (also categorised by age and anatomical location of IF among patients). All the data were in number or percentage. Results: Total 49 patients had IF with an overall prevalence of 0.82%. Male: female ratio was 1.04:1. Mean age of patients was 47.6±29.03 years. Majority 29 (59.18%) patients were in age group 31-60 years. Most common IF noted was meningioma 13 (26.5%) and 10 out of 13 meningioma patients belonged to 31-60 years age group. Other findings like glioma and pituitary macroadenoma were noted in 5 (10.2%) patients each. Total 22 (44.89%) patients had IF in supratentorial region of brain. Conclusion: Overall, low prevalence of IF was noted in the present study. Meningioma was one of the most common findings, especially among adults. Review of MRI scans by neuroradiologist, neurophysician and neurosurgeon for diagnosis of IF should be made mandatory. [ABSTRACT FROM AUTHOR]

Background: With the incorporation of molecular subtyping in glioma patients in 2016 WHO classification, there is a need to understand the immunohistochemistry (IHC) marker expression in various glioma patients and to clinically correlate with various subgroups. Objective: Aim of the study was to assess IHC marker expression profile in glioma patients and to clinically correlate them in various subgroups. Materials and Methods: The prospective study included 115 glioma patients. IHC markers (isocitrate dehydrogenase [IDH] 1, ATRX, P53, Ki-67 antibody) were done in all patients. Patients received treatment as per the grade of tumor. The patients were followed in 3 monthly intervals, for a period of 12 months. SPSS software version 20.0 was used for statistical analysis. Tables were prepared in Microsoft Excel sheet. Kaplan-Meier method was used for survival analysis. Results: There were 11 Grade 1, 33 Grade 2, 26 Grade 3, and 45 glioblastoma multiforme (GBM) patients out of which 10 patients were secondary GBM cases. IDH1 mutation is frequent in Grade 2 and Grade 3 tumors of both astrocytic and oligodendroglia tumors. Its mutation is also common in secondary GBM patients. ATRX mutation is specific to astrocytic lineage, Grade 2, Grade 3, and secondary GBM patients. Conclusion: Molecular nature of DA and AA cases can be accurately confirmed by combined IDH1 and ATRX IHC thereby avoiding costly investigations such as fluorescence in situ hybridization. In astrocytic tumors, p53 can act as a surrogate marker. IDH-mutant glioma patients have better prognoses than IDH wild gliomas. [ABSTRACT FROM AUTHOR]

Background: Neuro-oncology is a relatively young subspecialty of neurosurgery. 2018 was the 10th year since the founding of the Indian Society of Neuro-oncology.Objective: To assess patterns in neuro-oncology practice in India.Methods: This was an online survey covering various domains of neuro-oncology such as demographics and practice setting, protocols for the medical management of patients with brain tumors, protocols for surgery and the perioperative period (including antibiotic prophylaxis, dural closure techniques, etc.), technological adjuncts used for brain/spine tumors (including intraoperative neurologic monitoring-IONM), and management protocols for certain specific clinical scenarios.Results: The response rate was 13%. Although 37% of the respondents' institutions could be considered as having reasonable surgical volumes (>1 procedure/day), only about half of these had high volumes of malignant brain tumor surgery. A wide variation was seen in medical management, perioperative protocols, use of adjuncts and intraoperative technologies, and paradigms for specific clinical scenarios.Conclusions: There is a need to standardize the protocols in neuro-oncology. This could be achieved by strengthening the formal training process in surgical neuro-oncology. [ABSTRACT FROM AUTHOR]

Context: Gliomas are a heterogeneous group of relatively rare cancers that have an important public health-care implication due to their high levels of mortality and morbidity. While standard management guidelines are available, their implementation in a resource-limited scenario needs greater scrutiny. Settings and Design: This is a retrospective analysis of disease characteristics, treatment parameters including the time to imaging and time to treatment, and overall survival (OS) at 1 and 5 years in patients of brain gliomas. Subjects and Methods: Demographic, clinical, and follow-up data of histologically proven glioma patients that received radiotherapy (RT) between 2009 and 2013 at two tertiary care hospitals of India were collected and analyzed. Statistical Analysis: Kaplan–Meier curves were used to compare OS at 12 and 60 months. Cross-tabulation and Pearson's Chi-square test were used to study the association of study variables with survival. Results: One hundred and nine patients were included. The mean age was 45 years and males were three times as common as females. Astrocytomas were the most common histology with Grade IV astrocytomas comprising 48% of the total. The OS at 12 and 60 months was 79.8% and 24%, respectively, for the entire cohort. The average time taken for brain imaging from onset of symptoms was 24 days, while the time to surgery and the time to start RT were 18 and 44 days, respectively. Old age and ability to tolerate treatment were shown to affect survival at 1 year from diagnosis, though tumor histology and grade had an apparent impact on long-term prognosis. Conclusions: Hospital registries are an important source of demographic and clinical information on less common cancers such as gliomas. Increasing awareness among the general public and sensitization of primary health-care apparatus are critical for early diagnosis and treatment. [ABSTRACT FROM AUTHOR]

Background: Gliomas comprises the group of most common primary tumour of central nervous system. The current study was undertaken to evaluate the various usual and unusual radiological presentations among the patients of glioma among the Indian population. Aims and Objective: The aim of the current study was to observe the various radiological presentations of glioma occurring among the subjects. It was also intended to correlate the radiological and histopathological grading in glioma among subjects. Materials and Methods: The current study was a prospective observational study carried out among seventy-five patients admitted in a tertiary care hospital in Eastern India with provisional diagnosis of supratentorial glioma and operative biopsy confirmed to be Glioma. Results: The most common grade of tumour encountered was grade IV tumour. Of the clinical features, 70.66 % patients had hypodense lesion, 25.33% patients had iso dense and hypodense (mixed density) lesion and 3 patients had hyperdense lesion on CT Brain. On MRI in T1 Weighted images, 81.33% patients had hypointense lesion lesion on MRI T1 image. On T2 Weighted Images, 76% patients had hyperintense lesion., 77.33% demonstrated heterogenous enhancement. Ring enhancement was seen in 9 cases Minimal to no enhancement was seen in 8 cases. MRS showed maximum cases had Choline peak with altered Choline: creatinine ratio and decreased NAA peaks. Conclusion: Good imaging interpretations is crucial for planning of surgical excision and adjuvant radiotherapy. This review has been put to solve the basic problem in interpreting the Ct Scan and MRI of Glioma. [ABSTRACT FROM AUTHOR]

Purpose: Delayed diagnosis and poor awareness are significant barriers to the early intervention of pediatric brain tumors. This multicenter observational study aimed to evaluate the baseline routes and time to diagnosis for pediatric brain tumors in Tamil Nadu (TN), with the goal of promoting early diagnosis and timely referrals in the future., Methods: A standard proforma was used to retrospectively collect information on demographics, diagnosis, referral pathways, and symptoms of incident pediatric brain tumor cases between January 2018 and October 2020 across eight tertiary hospitals in TN. Dates of symptom onset, first presentation of health care, and diagnosis were used to calculate total diagnostic interval (TDI), patient interval (PI), and diagnostic interval (DI)., Results: A total of 144 cases (mean age, 6.64 years; range, 0-15.1 years) were included in the analysis. Among those, 94% (135/144) were from city/district areas, 40% (55/144) were self-referred, and 90% (129/144) had one to three health care professional visits before diagnosis. Median TDI, PI, and DI were 3.5 (IQR, 1-9.3), 0.6 (IQR, 0.1-4.6), and 0.6 (IQR, 0-3.3) weeks, respectively. Low-grade gliomas had the longest median TDI (6.6 weeks), followed by medulloblastomas (4.6 weeks) and high-grade gliomas (3.3 weeks). Average number of symptoms recorded was 1.7 at symptom onset and 1.9 at diagnosis., Conclusion: Although there are some similarities with data from the United Kingdom, many low-grade and optic pathway tumors were unaccounted for in our study. DIs were relatively short, which suggests that infrastructure may not be a problem in this cohort. Increased training and establishment of proper cancer registries, combined with proper referral pathways, could enhance early diagnosis for these children.

Introduction: With improvements in overall and progression-free survival in gliomas, current focus in neurosurgical oncology has largely shifted to the issue of quality of life (QOL) after treatment. There are not too many prospective studies evaluating QOL in these patients. We prospectively analyzed the health related quality of life (HRQOL) using the short form (SF) 36 questionnaire among patients harbouring a supratentorial glioma who underwent surgery at a tertiary care center in India.Methods: HRQOL was evaluated prospectively in 103 patients, between May 2016 and November 2017, at Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow. Score improvements were evaluated in terms of the smallest difference that a patient perceived to be clinically beneficial. The responsiveness of scores was assessed as minimum clinically important difference (MCID), effect size (ES) and relative efficiency. Cronbach's alpha co-efficient was used to assess the reliability of the result obtained.Results: The mean pre-operative score was the highest in the mental health domain (50.21±21.45) and significantly lower in patients with raised intracranial pressure (ICP), pre-operative deficits, a poor Karnofsky performance scale (KPS) score, in large tumors (>4cm) located in an eloquent location and in deep-seated tumors. A significant improvement was noted across all domains except the physical role (P-value 0.20). Overall, the general health domain was most responsive to change with the largest t-value (6.56) and 55.7% patients achieved their target MCID in general health (GH) domain. Among all the factors studied, only a low baseline SF-36 score was significantly associated with a change in QOL after surgery.Conclusion: Most parameters of HRQOL improved following surgery. Only small-to-moderate improvements occured in the early follow-up period. Large improving trends were noted on a long-term basis, irrespective of the histo-pathological grade. Similar improvements were also noted in patients who subsequently underwent re-surgery despite the development of post-operative complications or new deficits. Also, a low baseline QOL score heralded a poor survival. [ABSTRACT FROM AUTHOR]

Purpose: Studies have shown that cellularity of glial tumors are inversely correlated to minimum apparent diffusion coefficient (ADC) values derived on diffusion-weighted imaging (DWI). The purpose of this prospective exploratory study was to evaluate whether temporal change in "minimum ADC" values during follow-up predict progressive disease in glial tumors post radiotherapy and surgery. Materials and Methods: Adult patients of glial tumors, subjected to surgery followed by Radiotherapy (RT), were included in the study. Serial conventional magnetic resonance imaging with DWI at the following time points - presurgery, pre-RT, post-RT imaging at 3, 7, and 15 months were done. For "minimum ADC" values, multiple regions of interest (ROI) were identified on ADC maps derived from DWI. A mean of 5 minimum ADC values was chosen as "minimum ADC" value. The correlation was drawn between histology and minimum ADC values and time trends were studied. Results: Fourteen patients were included in this study. Histologies were low-grade glioma (LGG) - 5, anaplastic oligodendroglioma (ODG) -5, and glioblastoma multiforme (GBM) - 4. Minimum ADC values were significantly higher in LGG and GBM than ODG. Presurgery, the values were 0.812, 0.633, and 0.787 × 10-3 mm2/s for LGG, ODG, and GBM, respectively. DWI done at the time of RT planning showed values of 0.786, 0.636, 0.869 × 10-3 mm-3/s, respectively. During follow-up, the increasing trend of minimum ADC was observed in LGG (P = 0.02). All these patients were clinically and radiologically stable. Anaplastic ODGs, however, showed an initial increase followed by the fall of minimum ADC in all the 5 cases (P = 0.00). Four of the five cases developed progressive disease subsequently. In all the 4 GBM cases, a consistent fall of minimum ADC values was observed (P = 0.00), and they all progressed in spite of RT. Conclusions: The DWI-derived minimum ADC values are an important yet simple quantitative tool to assess the treatment response and disease progression before they are evident on conventional imaging during the follow-up of glial tumors. [ABSTRACT FROM AUTHOR]

Objective: In recent decades, a significant body of research has focused on targeted therapies for the treatment of central nervous system (CNS) tumors to enhance the effectiveness of management strategies. However, most of these efforts have been centered in high-income countries, which renders the generalizability of their results to low- and middle-income countries questionable. Therefore, in this review, we systematically investigated the status of research conducted on targeted therapy for CNS tumors in low- and lower-middle-income countries to elucidate the contribution of these countries in advancing neuro-oncology., Methods: A systematic search of 3 databases was performed using a predefined search strategy. After screening the articles based on our inclusion/exclusion criteria, the data were extracted to a predesigned Excel worksheet., Results: A review of 44 included studies showed that India, Iran, and Lebanon were the only countries with a contribution to this field. All included studies were laboratory or animal experiments, and there were no clinical studies in this field. The most investigated CNS tumor was malignant glioma, and gene-targeted therapy was the most investigated category of targeted therapies in these countries., Conclusions: Low- and lower-middle-income countries comprise more than half of the world population, but they are deprived of targeted therapies against CNS tumors. Although there are basic experiments performed on this subject, they originate in a limited number of these countries. Therefore, targeted therapy is in its preliminary stage in these countries., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Introduction: Diffusion weighted imaging and diffusion tensor imaging (DTI) give information about the amount and directionality of water diffusion occurring in a given tissue. Here we study the role of diffusion tensor metrics including fractional anisotropy (FA) and spherical anisotropy (CS) in preoperative grading of diffusely infiltrating astrocytomas. Methods: We performed DTI in 38 patients with pathologically proven diffusely infiltrating astrocytomas, who were classified into two groups, i.e., 15 patients with high-grade astrocytoma (HGAs, WHO grade III and IV) and 23 patients with low-grade astrocytoma (LGAs, WHO grade II). We measured maximum FA and minimum CS values in all cases from tumor. Histopathological diagnosis was established in all cases. Results: The mean maximum FA values were higher in HGA (0.583 ± 0.104) than LGA (0.295 ± 0.058), while mean minimum CS values were lower in HGA (0.42 ± 0.121) than LGA (0.722 ± 0.061). The difference in the diffusion tensor indices between HGA and LGA was found to be statistically significant with P value of <0.001. Keeping cutoff FA value of 0.4, all HGAs showed higher maximum FA values, and all LGAs showed lower maximum FA values. Also, all HGAs showed minimum CS values less than a cutoff value of 0.6, and all LGAs showed minimum CS values higher than 0.6. Conclusion: Diffusion tensor metrics such as maximum FA and minimum CS can help to differentiate HGA from LGA. [ABSTRACT FROM AUTHOR]

Background: FAT1 is a transmembrane protein and function as an adhesion molecule. FAT1 gene is located on chromosome 4q35.2 encoding a 506KDa protein. The role of FAT1 in tumors is not fully characterized. Few reports suggest it behaves as a tumor suppressor and few reported it as oncogenic. Here in our study we are characterizing the role of FAT1 in primary brain tumors. There are studies showing altered expression of FAT1 in human gliomas with ~30% of them showing LOH at the FAT1 locus. TP53 is mutated in 50% of primary brain tumors. P53 being a transcription factor, regulates the expression of a number of target genes including regulating biological functions such as cell cycle arrest, apoptosis, senescence, differentiation and antiangiogenesis. Glioma is known to have hypoxic environment in its tumour core and hypoxic tumour micro environment and HIF1A is believed to play role in tumour progression . So, in this study we are analysing the interaction between FAT1, P53 and HIF1A in glioma progression. Methodology: In order to delineate molecular crosstalk among factors driving glioma progression, we used knockdown and overexpression strategies in glioma cell lines followed by expression analysis by Q-PCR, Immunofluorescence and Western Blot. Migratory potential of cells was assessed by Migration assay and invasion potential were assessed using Matrigel invasion assay and images were captured using Nikon fluorescence microscopy and confocal microscopy. For cell proliferation and viability assay MTT/ XTT were used. Cell cycle analysis was carried out using PI staining in Flow cytometry (BD canto FACS) Sample Collection: - Glial tumor samples were collected from Department of Neurosurgery/Pathology of All India Institute of Medical Sciences, India. Tissue were collected in 1 ml of RNA-Later solution and incubated for 24 hours at 4°C. After removing RNA-Later solution the tumor tubes were immediately stored at -70°C till further use. For the control, normal human brain RNA was purchased commercially. Transfection of cell lines: - 1) For Knockdown: Selection of target siRNA and corresponding control- We have been using commercially available siRNA (Invitrogen) for FAT1 knockdown in our lab. FAT1 and control siRNA Oligos are designed by Genscript siRNA construct builder software(http://www.genscript. com/siRNA_construct_builder.html). 2) FAT1_trunc (Truncated FAT1) transfection: plasmid containing truncated FAT1 gene in pcDNA vector 3.1 was used (gifted by Dr Timothy A Chan). Truncated FAT1 plasmid contains first two repeats of cadherins at N terminals and around 22 repeats of cadherins are removed retaining last few cadherin repeats along with intercellular region of FAT1 gene. The vectors is tested for its expression and also known to retain all its functions. Transfection is carried out for 48 hr. Empty vector PCDNA is used as control. Transfection was carried out after plating 40000 to 50000 cells in 6 well plates for U87MG cells line and 1.2-1.5 lakhs cells for GOS and U373 Cell lines. The total duration of transfection was 72 hr (excluding plating) for knockdown and 48 hr for overexpression experiments. Hypoxia exposure was given with 0.2% oxygen for 48 hr After transfection following experimental techniques were done • MTT/XTT assay - For Cell survival • Flow cytometry- For Apoptosis assay/cell cycle analysis • RNA isolation and lysate preparation- For pathway(s) analysis by Real-Time PCR and Western blotting. • Migration and invasion assay - BD Matrigel • ICC Results Hypoxic treatment exhibited a significant increase in expression levels (7 to 9-fold) of FAT1 in U87MG, U373MG and GOS3 glioma cell lines. On FAT1 Knockdown under severe hypoxia, P53 and HIF1A expression was decreased and on Fat1 Knockdown under normoxia the expression of p53 and HIF1A was increased. The downstream targets of P53 (Noxa, Puma, p21 and MDM2) and downstream targets of HIF1A (VEGF, Ca9, GLUT1 and PGK3) also showed similar trend as p53 and HIF1A. On western blot analysis, protein levels of HIF1A and p53 showed increased levels after FAT1 knockdown under normoxia and no significant change was observed under FAT1 Knockdown in hypoxia. Our array data showed differential gene expression on FAT1 after knockdown. We observed reduction in HIF1A expression after FAT1 Knockdown under hypoxia and increased p53 and HIF1A on FAT1 Knockdown under normoxia. Increased in FAT1 expression under hypoxia and reduction of HIF1A after FAT1 Knockdown seems to suggest a direct link of FAT1 and HIF1A. Further, on subjecting U87 MG (grade IV GBM cell line) to hypoxia, we observed the shift in subcellular localization of FAT1 from perinuclear and plasma membrane location to inside the nucleus on ICC experiments suggesting FAT1 itself act as a transcription factors or help other transcription factors to increase or decrease gene expression involved in hypoxia and tumour progression. On over expressing FAT1 trunc we observed increased p53 [removed]1.49 fold) in normoxia and 1.87 fold increase under hypoxia (0.2%O2). MDM2 (downstream target gene of p53) also increased after FAT1 over expression by 1.629 fold in normoxia and 1.42 in hypoxia. We also observed elevated HIF1A in U87 and GOS3 along with their downstream molecules. RNA was isolated from histopath positive glioma grade IV tumors samples and gene expression analysis was performed in 18 tumour samples. We observed in GBM patients samples a positive correlation between FAT1 expression and P53 (r2 0.723 p=0.0029) and positive correlation of p53 with HIF1A (r2 =0.620, p=0.0106) and correlation of FAT1 expression with HIF1A showed r2 0.44, p=0.063. The expressions correlating analysis for patient samples and qPCR analysis of glioma cell lines suggests an existence of an intermediary molecule connecting the HIF1A and p53 regulation by FAT1. We examined p300 a co transcription factor necessary for both p53 and HIF1A transcription activity. P300 is upregulated when FAT1 is knockdown under normoxic condition. Absence of FAT1 (knockdown) under normoxia in U87MG cell lines increased p53 (1.7fold) and HIF1A (1.5 fold) and also p300 (1.8 fold). Increase in P53 in FAT1 Knockdown under normoxia is due to C/EBP beta. C/EBP is a known transcription factor and has two binding site in p53 promotors and C/EBP beta expressions is increased on FAT1 knockdown under normoxia to 1.56 fold. HIF1A being onocogene should promotes proliferation/migration and invasion but in our study we observed decreased in migration and invasion (Matrigel assay) and decreased in proliferation (MTT/XTT cell proliferation assay) and increased in apoptosis (Annexin FITC and PI flowcytometry assay) and increased Sub G0 population (PI cell cycle assay) on Knock down of FAT1 in normoxia. Increased p53 and its downstream genes after FAT1 Knockdown activated p300/p53 combination over p300/HIF1A resulting in decreased oncogenicity in normoxia in absences of FAT1. On other hand after FAT1 over expressions we observed decreased p300 and decreased p53 and its downstream and increased migration/ invasion and viability. The cell viability was 74.61 % after FAT1 over expression compared to control under normoxia. In hypoxia, the cell viability was increased after FAT1 over [removed]91% compared to control). Migratory properties also showed significant increase in over expression sample (4.19 times) then mock control under normoxia. This observation points to the critical and important role of FAT1 in cell migration. In conclusion FAT1 plays an oncogenic role in glioma and HIF1A and p53 plays and important role. The exact mechanism involved in the oncogenesis by FAT1 is under study. [ABSTRACT FROM AUTHOR]

Unlabelled: Purpose of the present study was to evaluate the role of (18)F-FDOPA PET/CT for predicting survival in patients with suspected recurrent glioma., Methods: A total of 33 previously treated, histopathologically proven glioma patients with clinical and contrast enhanced MRI findings suspicious for recurrence were enrolled in this prospective study. All patients underwent (18)F-FDOPA PET/CT. Ratios of tumor uptake to normal tissue uptake were generated by dividing the tumor SUVmax with SUVmax of the contralateral normal brain tissue (T/N), normal striatum (T/S), normal white matter (T/W) and normal cerebellum (T/C). Patients were followed up clinically and by repeated imaging. Data was censored, if the patient died of disease or at the end of the study. Survival analysis was performed for the distributions of each variable and by multivariate analysis., Results: (18)F-FDOPA PET/CT was positive for recurrence in 25 patients and negative in 8. Death occurred in nineteen patients. Median follow up period was 20.2 months. Median survival in this study was 39.2 months. In univariate analysis significant association of survival was noted with results of (18)F-FDOPA PET/CT (P=0.007) and (18)F-FDOPA PET/CT quantitative parameters namely SUVmax (P=0.001), T/S (P=0.005), T/W (P=0.0004), T/N (P=0.001) and T/C (P=0.003) were found to be significant. On multivariate analysis, only MRI size of the recurrent tumor (P=0.002) and T/N ratio of (18)F-FDOPA PET/CT (P=0.005) were found to be independent predictors of survival., Conclusion: T/N ratio on (18)F-FDOPA PET/CT is an independent predictor of survival in patients with suspected recurrent glioma, along with size of recurrent tumor on MRI., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)