ZnO nanoparticles-associated mitochondrial stress-induced apoptosis and G2/M arrest in HaCaT cells: a mechanistic approach.

Zinc oxide nanoparticles (ZnO NPs) with their wide range of consumer applications in day-to-day life received great attention to evaluate their effects in humans. This study has been attempted to elucidate the DNA damage response mechanism in a dermal model exposed to ZnO NPs through Ataxia Telangiectasia Mutated (ATM)-mediated ChK1-dependent G2/M arrest. Further, viability parameters and mechanism involved in the cell death with special reference to the consequences arising due to DNA damage were explored. Our study showed that ZnO NPs at concentrations 5 and 10 µg/ml induced significant cytotoxic effect in skin cell line. Moreover, the results confirmed generation of reactive oxygen species (ROS) induces the cell death by genotoxic insult, leading to mitochondrial membrane depolarisation and cell cycle arrest. Subsequently, ZnO NPs treatment created DNA damage as confirmed via Comet assay (increase in olive tail moment), micronucleus assay (increase in micronucleus formation), double-strand breaks (increase in ATM and Ataxia Telangiectasia and Rad3 related (ATR) expression), DNA fragmentation and cell cycle (G2/M arrest) studies. Finally, marker proteins analysis concluded the mechanistic approach by demonstrating the key marker expressions HMOX1 and HSP60 (for oxidative stress), cytochrome c, APAF1, BAX, Caspase 9, Caspase 3 and decrease in BCL2 (for activating apoptotic pathway), pATM, ATR and γH2AX (for double-strand breaks), DNA-PK (involved in DNA repair) and decrease in cell cycle regulators. In together, our data revealed the mechanism of ROS generation that triggers apoptosis and DNA damage in HaCaT cell lines exposed to ZnO NPs.

A new systematic approach of teaching and learning of forensic science for interdisciplinary students: A step towards renovating the forensic education system.

The new pedagogical approach of teaching and learning provides better deliverables by the teachers and better understanding and student engagement. In this order, a course was designed on Forensic Science for undergrad students from interdisciplinary background. Six pedagogies were used in this course with the aim to develop creativity, critical and logical thinking, practical learning, social accountability and research aptitude among the students. The main reasons for selecting these students were to avoid influencing the learning outcome due to their prior knowledge and observing the actual impact of the pedagogical learning. The suggested approach may be helpful in reducing the theoretical and practical gap in forensic science education. Besides, this teaching and learning approach may open a new avenue of forensic research and may result in a paradigm shift.

Forensic application of comet assay: an emerging technique.

Postmortem interval (PMI) estimation is a recurring problem in the field of forensic medicine. Conventional methods are effective but are insufficient to estimate accurate and precise time of death or PMI. In addition, degradation of biological samples is another major problem in forensic science which affects the investigation process and misleads the result. Some previous studies reported that DNA fragmentation has strong correlation with PMI. DNA fragmentation increased with prolonged PMI. Comet assay is a rapid sensitive, versatile, reliable and cost effective technique that is specifically used for qualitative and quantitative estimation of nuclear DNA fragmentation. Due to this attribute, comet assay can help to estimate accurate and precise time of death for some extent that is for early PMI estimation. In addition, two confounding factors are responsible for DNA fragmentation: (1) micro-organism; (2) environmental condition. Here, comet assay plays a dual role: (1) partially degraded samples get repaired using repair enzyme; (2) accurate time since deposition can be measured without using repair enzyme. Furthermore, this assay can also help to identify potential exposures of environmental-released chemicals/toxicants and its deleterious effects on human population. In this way, comet assay shows its versatile applications that could be useful for forensic investigation. Therefore, with the help of this review, an attempt was made to explore the versatility of comet assay technique for forensic applications and its future perspective.