Raj Kamal Singh

A high-throughput in vitro drug screen in a genetically engineered mouse model of diffuse intrinsic pontine glioma identifies BMS-754807 as a promising therapeutic agent.

Diffuse intrinsic pontine gliomas (DIPGs) represent a particularly lethal type of pediatric brain cancer with no effective therapeutic options. Our laboratory has previously reported the development of genetically engineered DIPG mouse models using the RCAS/tv-a system, including a model driven by PDGF-B, H3.3K27M, and p53 loss. These models can serve as a platform in which to test novel therapeutics prior to the initiation of human clinical trials. In this study, an in vitro high-throughput drug screen as part of the DIPG preclinical consortium using cell-lines derived from our DIPG models identified BMS-754807 as a drug of interest in DIPG. BMS-754807 is a potent and reversible small molecule multi-kinase inhibitor with many targets including IGF-1R, IR, MET, TRKA, TRKB, AURKA, AURKB. In vitro evaluation showed significant cytotoxic effects with an IC50 of 0.13 μM, significant inhibition of proliferation at a concentration of 1.5 μM, as well as inhibition of AKT activation. Interestingly, IGF-1R signaling was absent in serum-free cultures from the PDGF-B; H3.3K27M; p53 deficient model suggesting that the antitumor activity of BMS-754807 in this model is independent of IGF-1R. In vivo, systemic administration of BMS-754807 to DIPG-bearing mice did not prolong survival. Pharmacokinetic analysis demonstrated that tumor tissue drug concentrations of BMS-754807 were well below the identified IC50, suggesting that inadequate drug delivery may limit in vivo efficacy. In summary, an unbiased in vitro drug screen identified BMS-754807 as a potential therapeutic agent in DIPG, but BMS-754807 treatment in vivo by systemic delivery did not significantly prolong survival of DIPG-bearing mice.

Disinfection of water by pulsed power technique: a mechanistic perspective

The mechanism of disinfection by pulsed plasma technology was investigated in detail, with Escherichia coli as the model bacteria. Studies were carried out to investigate the behavior of bacterial surface potential as a function of applied energy, based on molecular level analysis of DNA and proteins. The cell survival and morphological changes during plasma treatment were correlated with membrane damage using SEM analysis, along with DNA degradation and protein leakage. In addition, cell size (Z-average) and poly-dispersive index (PI) of E. coli cells were measured to understand the colloidal behavior of disinfected bacterial cells in plasma reactors. Significant membrane deformities and morphological changes were observed under SEM analysis, and lactate dehydrogenase (LDH) assay confirmed a drastic increase in membrane permeability upon plasma treatment. An energy dependent DNA fragmentation and protein leakage confirmed the killing of bacterial cells. The E. coli cell membrane zeta potential (−14.81 ± 2.08 mV) was neutralized to −1.08 ± 0.83 mV in 6 min of treatment time due to the propensity of H+ ions on bacterial membranes. The FTIR confirmed the changes occurring in the functional groups of bacterial cell membranes, which further emphasized the possible interaction with ROS/RNS and membrane damage.

Terrestrial Macrofungal Diversity from the Tropical Dry Evergreen Biome of Southern India and Its Potential Role in Aerobiology

Macrofungi have long been investigated for various scientific purposes including their food and medicinal characteristics. Their role in aerobiology as a fraction of the primary biological aerosol particles (PBAPs), however, has been poorly studied. In this study, we present a source of macrofungi with two different but interdependent objectives: (i) to characterize the macrofungi from a tropical dry evergreen biome in southern India using advanced molecular techniques to enrich the database from this region, and (ii) to assess whether identified species of macrofungi are a potential source of atmospheric PBAPs. From the DNA analysis, we report the diversity of the terrestrial macrofungi from a tropical dry evergreen biome robustly supported by the statistical analyses for diversity conclusions. A total of 113 macrofungal species belonging to 54 genera and 23 families were recorded, with Basidiomycota and Ascomycota constituting 96% and 4% of the species, respectively. The highest species richness was found in the family Agaricaceae (25.3%) followed by Polyporaceae (15.3%) and Marasmiaceae (10.8%). The difference in the distribution of commonly observed macrofungal families over this location was compared with other locations in India (Karnataka, Kerala, Maharashtra, and West Bengal) using two statistical tests. The distributions of the terrestrial macrofungi were distinctly different in each ecosystem. We further attempted to demonstrate the potential role of terrestrial macrofungi as a source of PBAPs in ambient air. In our opinion, the findings from this ecosystem of India will enhance our understanding of the distribution, diversity, ecology, and biological prospects of terrestrial macrofungi as well as their potential to contribute to airborne fungal aerosols.

Seasonal variation of the dominant allergenic fungal aerosols – One year study from southern Indian region

Quantitative estimations of fungal aerosols are important to understand their role in causing respiratory diseases to humans especially in the developing and highly populated countries. In this study we sampled and quantified the three most dominantly found allergenic airborne fungi, Aspergillus fumigatus, Cladosporium cladosporioides, and Alternaria alternata from ambient PM10 samples using the quantitative PCR (qPCR) technique in a southern tropical Indian region, for one full year. Highest concentrations of A. fumigatus and C. cladosporioides were observed during monsoon whereas A. alternata displayed an elevated concentration in winter. The meteorological parameters such as temperature, relative humidity, wind speed, and precipitation exhibited a substantial influence on the atmospheric concentrations of allergenic fungal aerosols. The morphological features of various allergenic fungal spores present in the PM10 were investigated and the spores were found to possess distinct structural features. In a maiden attempt over this region we correlate the ambient fungal concentrations with the epidemiological allergy occurrence to obtain firsthand and preliminary information about the causative fungal allergen to the inhabitants exposed to bioaerosols. Our findings may serve as an important reference to atmospheric scientists, aero-biologists, doctors, and general public.