Anu Raghunathan

The size and subcellular distribution of the cellular NADH pool governing sensitivity to mitochondrial perturbation remain poorly understood in glioblastoma. We propose that two factors, an absolute NADH pool-size threshold of 0.677 pmol/cell, at or below which the electron transport chain (ETC) is the sole route for NAD⁺ regeneration, and a substrate-dependency context above that value, together shape Complex I sensitivity across a 271-fold pharmacological gradient in three glioblastoma states. Using NAD⁺/NADH measurements, Seahorse profiling, IC₅₀ assays, and compartment-specific NADH oxidase (LbNOX), we show that the neural stem-like side population sits at this threshold, hypersensitive to rotenone and NADH oxidation. Above 0.677 pmol/cell, sensitivity is context-dependent: glutaminolytic LN229 is sensitive, whereas U87MG resists through an ETFDH-mediated Complex I bypass. Cytosolic LbNOX rescues or sensitises cells according to their baseline NAD⁺/NADH ratio. The ratio alone does not predict vulnerability; metabolic architecture does, providing a framework for subpopulation-specific ETC vulnerability in glioblastoma.

The production of nonribosomal peptides (NRPs) in non-native hosts remains challenging due to the size and complexity of the biosynthetic enzymes. Here, we report the first successful reconstruction of the δ-(L-α-aminoadipyl)-l-cysteinyl-d-valine (ACV) pathway in Pichia pastoris. By introducing codon-optimized versions of the pcbAB and npgA genes directly into the genome, we were able to rebuild the minimal pathway required for ACV synthesis, which we confirmed using LC–MS/MS. Although the starting strain produced only small amounts of ACV (∼5 ng/mL), yields increased 10-fold after adaptive evolution (∼50 ng/mL) and further improved with precursor supplementation (∼60 ng/mL). Transcriptomic analysis showed that the engineered strain reorganized its metabolism to meet the energy and precursor demands of NRPS activity. Overall, this study suggests that P. pastoris can serve as a practical, proof-of-concept host for complex NRP biosynthesis and lays the groundwork for future engineering of β-lactam pathways.

The grand challenge in biobased Manufacturing Lies in achieving the sustainable, economically competitive conversion of renewable biomass into high-value Chemicals capable of replacing fossil-derived products. Among these, 3-hydroxypropionic acid (3-HP) has emerged as a top-tier target—an exceptionally versatile platform molecule. It finds applications in the synthesis of acrylic acid, 1,3-propanediol, and other derivatives, positioning it as a potential cornerstone for bio-based plastics. This review consolidates the latest breakthroughs in microbial 3-HP production, encompassing advanced strain engineering, pathway rewiring, cofactor optimization, metabolic modeling, and flux balance analysis. We critically examine strategies to overcome inherent metabolic and physiological constraints, including byproduct suppression, redox balancing, and tolerance engineering. Emerging approaches—such as dynamic regulation of metabolic flux, control of cell morphology and density, and integration of co-production pathways—are highlighted for their capacity to boost yields and process robustness. Additionally, we address the fermentation process innovations targeting enhanced productivity, substrate efficiency, minimal nutrient input, and industrially relevant titres. Collectively, these insights Chart a clear path toward the scalable, sustainable biomanufacturer of 3-HP. Graphical abstract

Summary: The modern response to pandemics, critical for effective public health measures, is shaped by the availability and integration of diverse epidemiological outbreak data. Genomic surveillance has come to the forefront during the coronavirus disease 2019 (COVID-19) pandemic at both local and global scales to identify variants of concern. Tracking variants of concern (VOC) is integral to understanding the evolution of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in space and time. Combining phylogenetics with epidemiological data like case incidence, spatial spread, and transmission dynamics generates actionable information. Here we discuss the genome surveillance done in Pune, India, through sequencing 10,496 samples from infected individuals and integrating them with multiple heterogeneous outbreak data. The rise and fall of VOCs along with shifting transmission dynamics in the time interval of December 2020 to March 2022 was identified. Population-based estimates of the proportion of circulating variants indicated the second and third peak of infection in Pune to be driven by VOCs Kappa (B.1.617.1), Delta (B.1.617.2), and Omicron (B.1.1.529) respectively. Integrating single nucleotide polymorphism changes across all sequenced genomes identified C (Cytosine) > T (Thymine) and G (Guanine) > T (Thymine) substitutions to dominate with higher rates of adaptive evolution in Spike (S), RNA-dependent RNA polymerase (RdRp), and Nucleocapsid (N) genes. Spike Protein mutational profiling during and pre-Omicron VOCs indicated differential rank ordering of high-frequency mutations in specific domains that increased the charge and binding properties of the protein. Time-resolved phylogenetic analysis of Omicron sub-lineages identified specific recombinant X lineages, XZ, XQ, and XM. BA.1 from Pune was found to be highly divergent by global sequence alignment and hierarchical clustering. Our “band of five” outbreak data analytics that includes the integration of five heterogeneous data types indicates that a strong surveillance system with comprehensive high-quality metadata was critical to understand the spatiotemporal evolution of the SARS-CoV-2 genome in Pune. We anticipate the use of such integrated workflows to be critical for pandemic preparedness in the future.

With more than 140 million people infected globally and 3 million deaths, the COVID 19 pandemic has left a lasting impact. A modern response to a pandemic of such proportions needs to focus on exploiting all available data to inform the response in real-time and allow evidence-based decision-making. The intermittent lockdowns in the last 13 months have created economic adversity to prevent anticipated large-scale mortality and relax the lockdowns have been an attempt at recovering and balancing economic needs and public health realities. This article is a comprehensive case study of the outbreak in the city limits of Pune, Maharashtra, India, to understand the evolution of the disease and transmission dynamics starting from the first case on March 9, 2020. A unique collaborative effort between the Pune Municipal Corporation (PMC), a government entity, and the Pune knowledge Cluster (PKC) allowed us to layout a context for outbreak response and intervention. We report here how access to granular data for a metropolitan city with pockets of very high-density populations will help analyze, in real-time, the dynamics of the pandemic and forecasts for better management and control of SARS-CoV-2. Outbreak data analytics resulted in a real-time data visualization dashboard for accurate information dissemination for public access on the epidemic’s progress. As government agencies craft testing and vaccination policies and implement intervention strategies to mitigate a second wave, our case study underscores the criticality of data quality and analytics to decode community transmission of COVID-19.

Metabolic reprogramming and its molecular underpinnings are critical to unravel the duality of cancer cell function and chemo-resistance. Here, we use a constraints-based integrated approach to delineate the interplay between metabolism and epigenetics, hardwired in the genome, to shape temozolomide (TMZ) resistance. Differential metabolism was identified in response to TMZ at varying concentrations in both the resistant neurospheroidal (NSP) and the susceptible (U87MG) glioblastoma cell-lines. The genetic basis of this metabolic adaptation was characterized by whole exome sequencing that identified mutations in signaling pathway regulators of growth and energy metabolism. Remarkably, our integrated approach identified rewiring in glycolysis, TCA cycle, malate aspartate shunt, and oxidative phosphorylation pathways. The differential killing of TMZ resistant NSP by Rotenone at low concentrations with an IC₅₀ value of 5 nM, three orders of magnitude lower than for U87MG that exhibited an IC₅₀ value of 1.8 mM was thus identified using our integrated systems-based approach.