The management of Degenerative Lumbar Spondylolisthesis through Ayurveda - A Case Report

Introduction: Back pain is the most common symptom of degenerative lumbar spondylolisthesis. This pain is typically worse with activities such as bending and lifting, and often eases when lying down. This condition occurs as a consequence of the general aging process in which the bones, joints, and ligaments in the spine become weak and less able to hold the spinal column in alignment. In Ayurveda, the complaints of Degenerative Lumbar spondylolisthesis can be effectively managed on the treatment principles of Gridhrasi and Katishula. Methodology: The present article deals with a case of diagnosed grade 1 Degenerative Lumbar spondylolisthesis of L4 and L5 and was advised for surgery. The Ayurvedic diagnosis of Gridhrasi was made. Management included internal medications and procedures like Kati Basti and Agni Karma. Results: The effectiveness of Ayurvedic management was assessed by Roland Morris's low back pain and disability questionnaire scale and Oswestry Low back disability questionnaire. On Roland-Morris's low back pain and disability questionnaire, the before treatment score was 20(83.33%) and at the time of discharge, it was reduced to 4 (16.66%). That means the patient showed an improvement of 66.67% on the Roland-Morris low back pain and disability questionnaire. Conclusion: The present case substantiates the effectiveness of classical Ayurvedic treatment in degenerative lumbar spondylolisthesis

Learning of bimanual motor sequences in normal aging

While it is well accepted that motor performance declines with age, the ability to learn simple procedural motor tasks appears to remain intact to some extent in normal aging. Here we examined the impact of aging on the acquisition of a simple sequence of bimanual actions. We further asked whether such learning results from an overall decrease in response time or is also associated with improved coordination between the hands. Healthy young and old individuals performed a bimanual version of the classic serial reaction time task. We found no learning deficit in older adults and noted that older subjects were able to learn as much as young participants. We also observed that learning in both groups was associated with an overall decrease in response time, but switch cost, the increase in response time when a switch in hands was required during sequence execution, did not decrease with learning. Surprisingly however, overall switch cost was lower in the older group compared to the younger subjects. These findings are discussed in the context of interactions between procedural and declarative memory, reduced interhemispheric inhibition and more symmetric cortical activation during motor performance in normal aging

Expression and purification studies on ATM kinase

by Kaushik Bhowmik, Rashmi Bhakuni and Sivapriya Kirubakara

High MDC1 expression in cervical cancer cells can affect the chemo and radio- therapeutic response as it inhibition leads to increased cell death

by Neeru Singh, Rashmi Bhakuni and Sivapriya Kirubakara

High MDC1 expression in cervical cancer cells can affect the chemo- and radiotherapeutic response as its depletion leads to increased cell death

Cervical cancer is the third most frequent cancer and common cause of death in women worldwide. The work presented identifies mediator of DNA damage checkpoint 1 (MDC1) as an important molecular target to increase sensitivity of cervical cancer cells to chemo or radiotherapy. MDC1 functions as an adaptor protein for recruitment and retention of many other DNA damage repair proteins in ataxia telangiectasia mutated (ATM) pathway for double-stranded DNA damage repair. It is reported to be highly expressed in cervical cancer cells. Also, its expression tends to increase with increase in malignancy. We have studied in detail MDC1 mRNA expression in three cervical cancer cell lines, HeLa, SiHa and CasKi, in response to various genotoxic stresses including some known inhibitors, UV exposure or gamma irradiation through quantitative PCR. The cellular response to the DNA damage resulted in increase in MDC1 expression, which declined with increase in treatment time period. Protein expression and activation by Western blotting with anti-MDC1 and anti-phosphoMDC1 antibody indicated a higher level of phosphorylated as compared to unphosphorylated MDC1. The significance of this increase in MDC1 expression was studied by generating stable cell lines knocked down for MDC1 expression. The modified cell lines were assessed for apoptosis through various assays, including flow cytometry, and showed greater cell death in response to DNA damage. In summary, high MDC1 expression can significantly affect chemo or radiotherapeutic response and its inhibition can improve sensitivity to these treatments.by Neeru Singh, Rashmi Bhakuni and Sivapriya Kirubakara

Validation of NFBD1 as a novel molecular target in cervical cancer therapy utilising advanced molecular approaches

by Neeru Singh, Rashmi Bhakuniand and Sivapriya Kirubakara

Evaluation of MBP tagged-hATR kinase domain catalytic activity with p53 Ser-15 phosphorylation

DNA damage response (DDR) pathways form an integral part of the body’s repair machinery, and ATR (ataxia-telangiectasia and Rad3-related kinase) protein is one of the key mediators in the DDR pathway that helps in maintaining genomic integrity. A growing body of evidence suggests that inhibition of ATR can help sensitize tumor cells to combinatorial treatment. However, specific ATR kinase inhibitors have largely remained elusive until now. Despite much interest in the protein for more than a decade, there has been little characterization of only the kinase domain, an essential target site for a variety of ATR inhibitors. Here, we report our findings for the bacterial expression, purification, and biological characterization of this potentially important recombinant kinase domain, which could further be considered for structure elucidation studies. Introduction of a solubility partner, i.e., maltose binding protein (MBP), at the N-terminus of the ATR kinase domain generated a soluble form of the protein, i.e., MBP-tagged hATR kinase domain (MBP-ATR-6X His), which was found to be catalytically active, as assessed by substrate p53 Ser-15 phosphorylation (EPPLSQEAFADLWKK). Our results also highlight the prospect of utilization of the overexpressed recombinant ATR kinase domain in characterization of kinase domain specific inhibitors.by Rashmi Bhakuni, Althaf Shaik and Sivapriya Kirubakara

Design, Synthesis, and Docking Studies of New Torin2 Analogs as Potential ATR/mTOR Kinase Inhibitors†

Targeting DNA damage and response (DDR) pathway has become an attractive approach in cancer therapy. The key mediators involved in this pathway are ataxia telangiectasia-mutated kinase (ATM) and ataxia telangiectasia-mutated, Rad3-related kinase (ATR). These kinases induce cell cycle arrest in response to chemo- and radio-therapy and facilitate DNA repair via their major downstream targets. Targeting ATP-binding site of these kinases is currently under study. Torin2 is a second generation ATP competitive mTOR kinase inhibitor (EC50 = 250 pmol/L) with better pharmacokinetic profile. Torin2 also exhibits potent biochemical and cellular activity against ATM (EC50 = 28 nmol/L) and ATR (EC50 = 35 nmol/L) kinases. In this study, eight new Torin2 analogs were designed and synthesized through multistep synthesis. All the synthesized compounds were characterized by NMR and mass analysis. The newly synthesized analogs were evaluated for their anti-cancer activity via CellTiter-Glo® assay. Additionally, compounds 13 and 14 also showed significant inhibition for ATR and mTOR substrates, i.e., p-Chk1 Ser 317 and p70 S6K Thr 389, respectively. Compounds 13 and 14 displayed promising anti-cancer activity with HCT-116 cell lines in the preliminary study. Further, a comparative model of ATR kinase was generated using the SWISS-MODEL server and validated using PROCHECK, ProSA analysis. Synthesized compounds were docked into the ATP-binding site to understand the binding modes and for the rational design of new inhibitors.by Althaf Shaik, Rashmi Bhakuni and Sivapriya Kirubakara

ATR and ATM: key regulators of genome integrity

by Rashmi Bhakuni, Vichitra Behel, Vijay Thiruvenkatam and Sivapriya Kirubakara

ATR specific novel therapeutics for cancer

by Althaf Shaik et.a