**Author details**

used for color detection or chemiluminescent detection. However, chemiluminescent detection is much more sensitive than color detection. Expression of a target is normalized to an

Different signaling cascade checkpoints and markers are commonly evaluated to understand the effect of the exposing agent on the cell through RT PCR and western blot analysis [30]. Some of these are highlighted. Akt, protein kinase B is involved in cell proliferation, transcription, migration and glucose metabolism. Caspases particularly, caspase 3 and caspase 9 indicate the progression of apoptosis. LC3B indicates the onset of autophagic processes. Hsp70 and Hsp90 indicate the heightened cellular responses of protein folding to external stresses. Expression of NFκB is an inflammatory response towards survival and cellular propagation.

As important it is to design and execute meaningful investigations in understanding the effects of toxic agents on cellular functions, it is also important to do them with statistical validation. Experiment results need to be accurate and consistent within the framework of the defined system. Reproducibility of protocols and outcomes are important in formulating further scope for any research. In this endeavor, several statistical tools can be employed which include the determination of average, standard deviation, p value and range of error. Especially with studying toxicity from nanoparticle exposure, the number of exposing agents is exponentially growing every year. As such it becomes time and cost consuming to exact the required number of experiments to test each particle for various routes of exposure. This necessitates the need for development of adequate and accurate prediction tools for toxicity. A raising technique today is the advent of quantitative structure activity-based relationship (QSAR) models [31]. A response curve is defined, and various physiochemical and biological parameters are gathered or determined and fitness to response is evaluated. For *in vitro*

tools help screen large numbers of particles for their toxic outcomes. This ensure only the most promising leads are further evaluated with actual we laboratory experiments, in pro-

Accurate risk assessment is an interdisciplinary approach, where acquiring information and processing is a never-ending ordeal. Therefore, an ethical yet smart approach is needed for studying risks associated with nanoparticles. Only the understanding of the mechanisms of toxicity can enable future endeavors towards strategies of safe nanomaterial design and stress

(correlation coefficient) of any such effort must be greater than 0.81. Such

internal control such as β- Actin, to override experimental errors.

**7. Statistical analysis of experimental data**

experiments the R2

104 Cell Culture

**8. Conclusion**

recovery solutions.

cess, saving resources, time and funds.

mTOR is directly or indirectly involved in the regulation of protein function.

Ansie Martin\* and Angshuman Sarkar

\*Address all correspondence to: ansiemartin@gmail.com

CMBL, Department of Biological Sciences, Birla Institute of Technology and Sciences, K K Birla Goa Campus, Sancoale, South Goa, India
