Detection of serum M-protein in acetonitrile precipitates by MALDI-TOF mass spectrometry: A novel, low-cost methodology.

BACKGROUND: Several studies have demonstrated the analytical sensitivity of MALDI-TOF mass spectrometry (MALDI-TOF MS) by immunoenrichment for M-protein analysis. We report the results of a novel, low-cost, reagent-based extraction process using acetonitrile (ACN) precipitation to enrich for κ and λ light chains which can be analysed by MALDI-TOF MS. METHODS: Institutional Ethics committee approval was obtained. Serum samples from patients with monoclonal gammopathy of undetermined significance (MGUS), multiple myeloma (MM), plasmacytoma, AL amyloidosis and Waldenström macroglobulinemia (WM) underwent ACN precipitation. The images obtained were overlaid on apparently healthy donor serum samples to confirm the presence of M-protein. A sample was considered positive for M-protein if there was a sharp or broad peak within the κ or λ mass/charge (m/z) range: m/z- [M + 2H]2+: 11,550-12,300 Da and λ m/z- [M + 2H]2+: 11,100-11,500 Da. Images were acquired at a m/z range of 10,000-29,000 Da. Corresponding serum protein electrophoresis (SPEP), serum immunofixation electrophoresis (IFE) and serum free light chain (sFLC) assay by nephelometry were performed for all the samples. RESULTS: Two-hundred-and-two serum samples were included in the study: MM- 184 (91%); AL amyloidosis- 2 (1%); plasmacytoma- 8 (4%); MGUS- 6 (3%) and WM- 2 (1%). All the SPEP positive samples were identified by MALDI-TOF MS. Out of 179 samples positive for M-protein by IFE, MALDI-TOF MS was positive in 176 samples (98%). Compared to IFE, the sensitivity and specificity of M-protein identification by MALDI-TOF MS were 98.3% and 52.2%, respectively. CONCLUSIONS: This study demonstrates the feasibility of qualitatively identifying M-protein without the need for antibody-based immunoenrichment, making the technique cost-effective.

Economic perspective of cancer treatment in India.

Financial constraints faced by the families play a vital role in cancer treatment refusal, non-adherence, and failure of the prescribed therapy. This review aims to give an insight into the economic perspective of cancer treatment in India, focusing on the accessibility and affordability of oncological drugs, and the move towards generics/biosimilars without compromising on the quality of the treatment. The monthly cost of a set of drugs available in India for the treatment of solid malignancies, approved after 2010 by the US FDA and the Drugs Controller General of India (DCGI) were calculated based on standard patient parameters. The information on the clinical trial, the monthly cost of treatment, and the availability of its equivalent have been compiled. Newer cancer drugs are approved based on surrogate endpoints, with a very modest prolongation of life, but the cost incurred can be unbearable. There is a considerable variation in costs between the innovator and the equivalent drugs, making the latter cost-effective. We have highlighted the importance of generics and biosimilars, as a cost-cutting strategy, in delivering state-of-art health care with a lesser chance of treatment abandonment: this will ensure that all patients have equal access to personalized medicine which are reliable, effective, and affordable for better curative, supportive, and palliative care.

TGFß C-509T, TGFß T869C, XRCC1 Arg194Trp, IKBα C642T, IL4 C-590T Genetic polymorphisms combined with socio-economic, lifestyle, diet factors and gastric cancer risk: A case control study in South Indian population.

BACKGROUND: Gastric cancer is worldwide the third major cause of cancer related death. Risk factors for gastric cancer includes Helicobacter pylori infection, gastric ulcer, less hygienic condition, use of tobacco, alcohol consumption, use of salted, smoked food, genetic alterations etc. In order to identify the risk factors associated with gastric cancer in South Indian population a case-control study involving 200 proven gastric cancer cases and 400 controls was conducted. METHODS: A structured questionnaire was used to interview all the subjects who participated in our study. Genotyping assay was performed using Taqman allelic discrimination assay for 5 Single Nucleotide Polymorphisms (SNPs)-TGFß C-509T, TGFß T869C, XRCC1 Arg194Trp, IkBα C642T and IL4C-590T. RESULTS: Odds Ratios (ORs) and 95% confidence intervals (CIs) were calculated using conditional logistic regression. Statistical analysis on socio-economic factors, lifestyle factors had showed that subjects from low socio economic status, use of tobacco and consumption of non-vegetarian food had increased risk of developing gastric cancer. Multi-factorial analysis for the SNPs adjusting for the risk factors obtained in this study showed that TGFΒ C-509T TT genotypes had four fold increased risk of gastric cancer (OR = 4.11, CI = 1.02-16.56) and TGFß T869C CC genotype had a decreased risk of gastric cancer (OR = 0.21, CI = 0.05-0.85). CONCLUSION: Economic status, tobacco use and food habits play a significant role in gastric cancer development. TT genotype for TGFß C-509T had an increased risk and CC genotype for TGFß T869C had a decreased risk of gastric cancer in south Indian population after adjusting for socio-economic factors and lifestyle factors.

Assessment of the radiation sensitivity of cervical cancer cell lines.

The aim of radiotherapy is to kill tumor cells in a primary tumor, in draining lymph nodes, and/or in small metastatic lesions. The response of tumor cells to radiation depends on the dose, an individual's radiosensitivity, the duration of radiation exposure (i.e., the timing), the fraction size, and the presence of other variables (e.g., chemotherapy). Sensitivity of the cells to radiation can be determined by cell proliferation and clonogenicity assays, which assess the ability of the cells to survive at low cell densities and to successfully initiate and sustain cell proliferation over time yielding viable colonies or clones after irradiation with a range of doses (0-10Gy). Apart from assessing the sensitivity of the cells to radiation, these assays are now being increasingly used to test for the effects of drugs/genes on the growth and proliferative characteristics of cells in vitro. Additionally, they are being used to determine the combinatorial effect of novel agents or inhibitors, which can modify the response to radiation for a favorable therapeutic outcome. The rates of cell survival and proliferation obtained from these assays help in identifying the most sensitive and resistant cell lines among particular cancer types. Because of their wide range of application, from identifying the most sensitive and resistant cell lines, to evaluating novel therapeutic agents, we describe here the basic steps involved in assessing the radiosensitivity of cervical cancer cell lines.