DMA, a Bisbenzimidazole, Offers Radioprotection by Promoting NFκB Transactivation through NIK/IKK in Human Glioma Cells

BACKGROUND: Ionizing radiation (IR) exposure often occurs for human beings through occupational, medical, environmental, accidental and/or other sources. Thus, the role of radioprotector is essential to overcome the complex series of overlapping responses to radiation induced DNA damage. METHODS AND RESULTS: Treatment of human glioma U87 cells with DMA (5- {4-methylpiperazin-1-yl}-2-[2'-(3, 4-dimethoxyphenyl)-5'-benzimidazolyl] in the presence or absence of radiation uncovered differential regulation of an array of genes and proteins using microarray and 2D PAGE techniques. Pathway construction followed by relative quantitation of gene expression of the identified proteins and their interacting partners led to the identification of MAP3K14 (NFκB inducing kinase, NIK) as the candidate gene affected in response to DMA. Subsequently, over expression and knock down of NIK suggested that DMA affects NFκB inducing kinase mediated phosphorylation of IKKα and IKKβ both alone and in the presence of ionizing radiation (IR). The TNF-α induced NFκB dependent luciferase reporter assay demonstrated 1.65, 2.26 and 3.62 fold increase in NFκB activation at 10, 25 and 50 µM DMA concentrations respectively, compared to control cells. This activation was further increased by 5.8 fold in drug + radiation (50 µM +8.5 Gy) treated cells in comparison to control. We observed 51% radioprotection in control siRNA transfected cells that attenuated to 15% in siRNA NIK treated U87 cells, irradiated in presence of DMA at 24 h. CONCLUSIONS: Our studies show that NIK/IKK mediated NFκB activation is more intensified in cells over expressing NIK and treated with DMA, alone or in combination with ionizing radiation, indicating that DMA promotes NIK mediated NFκB signaling. This subsequently leads to the radioprotective effect exhibited by DMA

Nations within a nation: variations in epidemiological transition across the states of India, 1990–2016 in the Global Burden of Disease Study

18% of the world's population lives in India, and many states of India have populations similar to those of large countries. Action to effectively improve population health in India requires availability of reliable and comprehensive state-level estimates of disease burden and risk factors over time. Such comprehensive estimates have not been available so far for all major diseases and risk factors. Thus, we aimed to estimate the disease burden and risk factors in every state of India as part of the Global Burden of Disease (GBD) Study 2016

Glutathione Transferase GSTπ In Breast Tumors Evaluated By Three Techniques

The glutathione transferases are involved in intracellular detoxification reactions. One of these, GSTπ, is elevated in some breast cancer cells, particularly cells selected for resistance to anticancer agents. We evaluated GSTπ expression in 60 human breast tumors by three techniques, immunohistochemistry, Northern hybridization, and Western blot analysis. There was a significant positive correlation between the three methods, with complete concordance seen in 64% of the tumors. There was strong, inverse relationship between GSTπ expression and steroid receptor status with all of the techniques utili zed. [n addition, there was a trend toward higher GSTπ expression in poorly differentiated tumors, but no correlation was found between tumor GSTπ content and DNA ploidy or %S-phase. GSTπ expression was also detected in adjacent benign breast tissue as well as infiltrating lymphocytes; this expression may contribute to GSTπ measurements using either Northern hybridization or Western blot analysis. These re sults suggest that immunohistochemistry is the method of choice for measuring GSTπ in breast tumors

When monoclonal antibodies are not monospecific: Hybridomas frequently express additional functional variable regions.

Monoclonal antibodies are commonly assumed to be monospecific, but anecdotal studies have reported genetic diversity in antibody heavy chain and light chain genes found within individual hybridomas. As the prevalence of such diversity has never been explored, we analyzed 185 random hybridomas, in a large multicenter dataset. The hybridomas analyzed were not biased towards those with cloning difficulties or known to have additional chains. Of the hybridomas we evaluated, 126 (68.1%) contained no additional productive chains, while the remaining 59 (31.9%) contained one or more additional productive heavy or light chains. The expression of additional chains degraded properties of the antibodies, including specificity, binding signal and/or signal-to-noise ratio, as determined by enzyme-linked immunosorbent assay and immunohistochemistry. The most abundant mRNA transcripts found in a hybridoma cell line did not necessarily encode the antibody chains providing the correct specificity. Consequently, when cloning antibody genes, functional validation of all possible VH and VL combinations is required to identify those with the highest affinity and lowest cross-reactivity. These findings, reflecting the current state of hybridomas used in research, reiterate the importance of using sequence-defined recombinant antibodies for research or diagnostic use

When monoclonal antibodies are not monospecific: Hybridomas frequently express additional functional variable regions.

Monoclonal antibodies are commonly assumed to be monospecific, but anecdotal studies have reported genetic diversity in antibody heavy chain and light chain genes found within individual hybridomas. As the prevalence of such diversity has never been explored, we analyzed 185 random hybridomas, in a large multicenter dataset. The hybridomas analyzed were not biased towards those with cloning difficulties or known to have additional chains. Of the hybridomas we evaluated, 126 (68.1%) contained no additional productive chains, while the remaining 59 (31.9%) contained one or more additional productive heavy or light chains. The expression of additional chains degraded properties of the antibodies, including specificity, binding signal and/or signal-to-noise ratio, as determined by enzyme-linked immunosorbent assay and immunohistochemistry. The most abundant mRNA transcripts found in a hybridoma cell line did not necessarily encode the antibody chains providing the correct specificity. Consequently, when cloning antibody genes, functional validation of all possible VH and VL combinations is required to identify those with the highest affinity and lowest cross-reactivity. These findings, reflecting the current state of hybridomas used in research, reiterate the importance of using sequence-defined recombinant antibodies for research or diagnostic use

Bay of Bengal intraseasonal oscillations and the 2018 monsoon onset

Author Posting. © American Meteorological Society, 2021. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Bulletin of the American Meteorological Society 102(10), (2021): E1936–E1951, https://doi.org/10.1175/BAMS-D-20-0113.1.In the Bay of Bengal, the warm, dry boreal spring concludes with the onset of the summer monsoon and accompanying southwesterly winds, heavy rains, and variable air–sea fluxes. Here, we summarize the 2018 monsoon onset using observations collected through the multinational Monsoon Intraseasonal Oscillations in the Bay of Bengal (MISO-BoB) program between the United States, India, and Sri Lanka. MISO-BoB aims to improve understanding of monsoon intraseasonal variability, and the 2018 field effort captured the coupled air–sea response during a transition from active-to-break conditions in the central BoB. The active phase of the ∼20-day research cruise was characterized by warm sea surface temperature (SST > 30°C), cold atmospheric outflows with intermittent heavy rainfall, and increasing winds (from 2 to 15 m s−1). Accumulated rainfall exceeded 200 mm with 90% of precipitation occurring during the first week. The following break period was both dry and clear, with persistent 10–12 m s−1 wind and evaporation of 0.2 mm h−1. The evolving environmental state included a deepening ocean mixed layer (from ∼20 to 50 m), cooling SST (by ∼1°C), and warming/drying of the lower to midtroposphere. Local atmospheric development was consistent with phasing of the large-scale intraseasonal oscillation. The upper ocean stores significant heat in the BoB, enough to maintain SST above 29°C despite cooling by surface fluxes and ocean mixing. Comparison with reanalysis indicates biases in air–sea fluxes, which may be related to overly cool prescribed SST. Resolution of such biases offers a path toward improved forecasting of transition periods in the monsoon.This work was supported through the U.S. Office of Naval Research’s Departmental Research Initiative: Monsoon Intraseasonal Oscillations in the Bay of Bengal, the Indian Ministry of Earth Science’s Ocean Mixing and Monsoons Program, and the Sri Lankan National Aquatic Resources Research and Development Agency. We thank the Captain and crew of the R/V Thompson for their help in data collection. Surface atmospheric fields included fluxes were quality controlled and processed by the Boundary Layer Observations and Processes Team within the NOAA Physical Sciences Laboratory. Forecast analysis was completed by India Meteorological Department. Drone image was taken by Shreyas Kamat with annotations by Gualtiero Spiro Jaeger. We also recognize the numerous researchers who supported cruise- and land-based measurements. This work represents Lamont-Doherty Earth Observatory contribution number 8503, and PMEL contribution number 5193.2022-04-0