Metabolic Variations between Low-Grade and High-Grade Gliomas-Profiling by 1H NMR Spectroscopy.

Altered cellular metabolism is one of the crucial hallmarks of glioma that deserves exploration, as the metabolites act as direct indicators of protein function and genetic variations. The current study focused on the metabolomic profiling of patients from whom glioma specimens were obtained for the identification of specific metabolites that could distinguish the low grade and high grade. In the current study, 1H NMR spectroscopy was carried out and the data were analyzed by partial least-squares discriminant analysis (PLS-DA) and orthogonal projection to latent structure with discriminant analysis (OPLS-DA). Pathway analysis was done to associate characteristic metabolites with the grades of sample using MetaboAnalyst 4.0 software based on the KEGG metabolic pathways database. Distinctive metabolic profiles among low- and high-grade gliomas with top 15 characteristic metabolites that could discriminate these grades were identified on the basis of their VIP scores from the OPLS-DA model. The major altered metabolic pathways include choline, taurine and hypotaurine, glutamate/glutamine, glutathione, and phenyl alanine/tyrosine, which were found to be consistent with the particular grade of a sample. Our study clearly demonstrated a characteristic metabolic profile of individual grades of glioma, suggesting that an altered metabolism is consistent with the specific grades of glioma appreciation and could lead to the development novel treatment strategies.

Connexin 30 mediated rewiring of glucose metabolism in rat C6 xenograft and grades of glioma.

Connexin 30 (Cx30), a tumour-suppressive gap junctional protein, impacts on insulin-like growth factor receptor 1-mediated progression and stemness of glioma. Of late, metabolic reprogramming, a recently adjudged hall mark of malignancy, could reasonably associated with the changes in gap junctional communication in glioma. This newly recognized hallmark of reprogramming of metabolism to maintain the rapid proliferation necessitates further probing to establish the stronger hall marks. Hence, the current study attempted to link the association between the expression of Cx30 with glucose uptake and glucose metabolism in glioma. We have transfected Cx30 in C6 glioma cells, characterized by a low level of intercellular communication and developed xenografts to study the status of glucose transporters (GLUTs), hexokinase 2 and Pyruvate dehydrogenase kinase 1 (PDK 1) along with human glioma tissues by RT-PCR and immunoblotting. The results showed a significant increase in the levels of GLUTs, hexokinase 2 and PDK 1 in C6-implanted rat xenografts and high grades compared to their respective controls, whereas Cx30-transfected C6-implanted rat xenograft and low grades show no significant change compared to that of controls supporting the association between Gap junctional communications and glucose metabolism. We strongly speculate the impact of Cx30 over the glucose metabolism that might provide therapeutic prospects and challenges for anti-glycolytic cancer therapy.

Aberrant neurotransmissional mRNAs in cerebral ganglions of rotenone-exposed Lumbricus terrestris exhibiting motor dysfunction and altered cognitive behavior.

Rotenone (ROT) was shown to affect cerebral ganglions (CGs) of Lumbricus terrestris as a pioneering observation in our earlier investigation. Though ROT is a well-known neurotoxin causing neurodegeneration (ND), the precipitation of movement dysfunction remains largely unknown. We have designed the current study to analyze motor abnormalities in worms by exposing them to different concentrations (0.0-0.4 ppm) of ROT for 7 days. GABA, cholinergic receptor, serotonin transporter (SERT), acetylcholine esterase (AchE), and dopamine-ß-hydroxylase that are well known for their involvement in neuromuscular junctions were investigated by qRT-PCR. Further, neuronal mitochondrial genes (cytochrome C oxidase-2, NADH deydrogenase-1, cytochrome-b) and actin-1 that are essential for regeneration and calreticulin (phagocytosis) were investigated. The levels of neurotransmitters, lipids, ATPase, neuronal behavior analyses, and fluorescence analysis (lipid droplets) were performed in CGs which showed significant variations at 0.3 ppm. Ultrastructural changes in lipid droplet and neuromelatonin were prominent in 0.3 ppm. Dose-dependent effect of ROT on behavior alteration and expression of m-RNAs studied suggested that at 0.3 ppm, it could deteriorate motor and cognitive functions. We predict that perhaps, by virtue of its effect on cerebral ganglionic genes and their neurotransmitting potential, ROT may cause morbidities that resemble features characteristic of hemiparkinsonic degeneration.

Pre-clinical analysis of changes in intra-cellular biochemistry of glioblastoma multiforme (GBM) cells due to c-Myc silencing.

Glioblastoma Multiforme (GBM) is an aggressive form of brain Tumor that has few cures. In this study, we analyze the anti-proliferative effects of a new molecule JQ1 against GBMs induced in Wistar Rats. JQ1 is essentially a Myc inhibitor. c-Myc is also known for altering the biochemistry of a tumor cell. Therefore, the study is intended to analyze certain other oncogenes associated with c-Myc and also the change in cellular biochemistry upon c-Myc inhibition. The quantitative analysis of gene expression gave a co-expressive pattern for all the three genes involved namely; c-Myc, Bcl-2, and Akt. The cellular biochemistry analysis by transmission electron microscopy revealed high glycogen and lipid aggregation in Myc inhibited cells and excessive autophagy. The study demonstrates the role of c-Myc as a central metabolic regulator and Bcl-2 and Akt assisting in extending c-Myc half-life as well as in regulation of autophagy, so as to regulate cell survival on the whole. The study also demonstrates that transient treatment by JQ1 leads to aggressive development of tumor and therefore, accelerating death, emphasizing the importance of dosage fixation, and duration for clinical use in future.

Asiaticoside, a trisaccaride triterpene induces biochemical and molecular variations in brain of mice with parkinsonism.

BACKGROUND: Parkinson's disease characterized by oxidative stress and mitochondrial damage in the pars compacta of substantia nigra remains a challenge to manage with an added disadvantage of side effects of L-levo dopa, the standard drug used for therapy. Thus, an alternative approach of utilizing natural components would be beneficial in the management of the disease. The present study was aimed to investigate the potential role of asiaticoside (As), a trisaccaride triterpene against1 - methyl 4 - phenyl 1,2,3,6 tetrahydropyridine (MPTP)-induced neurotoxicity in experimental mice. METHODS: Mice were divided into 4 groups: Group I received vehicle saline, group II was treated with 20 mg/kg of body weight of MPTP (2 doses with 2 h intervals), group III received MPTP along with 50 mg/kg body weight of As for the 21 consecutive days starting from the day of MPTP intoxication. Group IV received 50 mg/kg body weight of asiaticoside for the same period serving as drug control. Animals were sacrificed at the end of experimental period and the striatum and midbrain samples were analyzed for enzyme assays, transmission electron microscopic (TEM) analysis. Immunofluorescent assay was performed to study the expression of GFAP to detect astrocyte, which are activated due to neuronal damage. Imunohistochemical studies were carried out to quantify the expression of Bax and Bcl2, the molecular signatures that would provide clues of the extent of neurodegeneration. RESULTS: The activities of enzymes were increased on As administration when compared with those of group II animals. Expressions of Bax and Bcl2 along with GFAP did show significant variations (p < 0.05) on MPTP treatment when compared to control animals and the changes were found to be reversed significantly (p < 0.05) after treatment with asiaticoside. TEM analysis also showed attenuated degenerative architecture on As administration. The mice which received As alone (drug control IV) did not show significant variation from that of the control mice. CONCLUSION: The observations suggest that asiaticoside may be efficacious in protecting neurons from the oxidative damage caused by the insult of MPTP.

Biochemical and cytogenetic analysis of brain tissues in different grades of glioma patients.

BACKGROUND: Glioma, a neoplasm of neuroglial cells, is the most common type of brain tumor, constituting more than 50% of all brain tumors. PURPOSE: This report summarizes the current knowledge regarding the clinical utility of biochemical enzyme markers for both diagnostic and therapeutic purposes in different grades of glioma. METHODS: Sixty patients with different grades of glioma include glioblastoma multiforme (n=20), Anaplastic astrocytoma (n=10). Oligodendroglioma (n=10). Ependymoma (n=10), Pilocytic astrocytoma (n=10) and patients with benign lesions (n=5) served as controls. Activity of antioxidants and marker enzymes were assayed in all grades followed by karyotyping. RESULTS: Activities of antioxidant enzymes reduced significantly (p<0.05) compared to controls except CAT, GST that showed marked increase (p<0.05) in accordance with malignancy. CK, Na-K(+) ATPases, 5'-Nucleotidases showed marked increase in grade IV. Similarly, Mg2-ATPase, Ca2+ATPases showed significant increase in grade III. CONCLUSION: The clinical importance for classification and treatment of glioma is governed by biochemical enzyme markers. The study of enzymes supported by related chromosomal changes is anticipated to provide better appreciation of biological properties in different grades of glioma.