Advancements in CHO metabolomics: techniques, current state and evolving methodologies.

Background: Investigating the metabolic behaviour of different cellular phenotypes, i.e., good/bad grower and/or producer, in production culture is important to identify the key metabolite(s)/pathway(s) that regulate cell growth and/or recombinant protein production to improve the overall yield. Currently, LC-MS, GC-MS and NMR are the most used and advanced technologies for investigating the metabolome. Although contributed significantly in the domain, each technique has its own biasness towards specific metabolites or class of metabolites due to various reasons including variability in the concept of working, sample preparation, metabolite-extraction methods, metabolite identification tools, and databases. As a result, the application of appropriate analytical technique(s) is very critical. Purpose and scope: This review provides a state-of-the-art technological insights and overview of metabolic mechanisms involved in regulation of cell growth and/or recombinant protein production for improving yield from CHO cultures. Summary and conclusion: In this review, the advancements in CHO metabolomics over the last 10 years are traced based on a bibliometric analysis of previous publications and discussed. With the technical advancement in the domain of LC-MS, GC-MS and NMR, metabolites of glycolytic and nucleotide biosynthesis pathway (glucose, fructose, pyruvate and phenylalanine, threonine, tryptophan, arginine, valine, asparagine, and serine, etc.) were observed to be upregulated in exponential-phase thereby potentially associated with cell growth regulation, whereas metabolites/intermediates of TCA, oxidative phosphorylation (aspartate, glutamate, succinate, malate, fumarate and citrate), intracellular NAD+/NADH ratio, and glutathione metabolic pathways were observed to be upregulated in stationary-phase and hence potentially associated with increased cell-specific productivity in CHO bioprocess. Moreover, each of technique has its own bias towards metabolite identification, indicating their complementarity, along with a number of critical gaps in the CHO metabolomics pipeline and hence first time discussed here to identify their potential remedies. This knowledge may help in future study designs to improve the metabolomic coverage facilitating identification of the metabolites/pathways which might get missed otherwise and explore the full potential of metabolomics for improving the CHO bioprocess performances.

Comparison of freeze-thaw and sonication cycle-based methods for extracting AMR-associated metabolites from Staphylococcus aureus.

Emerging antimicrobial resistance (AMR) among Gram-positive pathogens, specifically in Staphylococcus aureus (S. aureus), is becoming a leading public health concern demanding effective therapeutics. Metabolite modulation can improve the efficacy of existing antibiotics and facilitate the development of effective therapeutics. However, it remained unexplored for drug-resistant S. aureus (gentamicin and methicillin-resistant), primarily due to the dearth of optimal metabolite extraction protocols including a protocol for AMR-associated metabolites. Therefore, in this investigation, we have compared the performance of the two most widely used methods, i.e., freeze-thaw cycle (FTC) and sonication cycle (SC), alone and in combination (FTC + SC), and identified the optimal method for this purpose. A total of 116, 119, and 99 metabolites were identified using the FTC, SC, and FTC + SC methods, respectively, leading to the identification of 163 metabolites cumulatively. Out of 163, 69 metabolites were found to be associated with AMR in published literature consisting of the highest number of metabolites identified by FTC (57) followed by SC (54) and FTC + SC (40). Thus, the performances of FTC and SC methods were comparable with no additional benefits of combining both. Moreover, each method showed biasness toward specific metabolite(s) or class of metabolites, suggesting that the choice of metabolite extraction method shall be decided based on the metabolites of interest in the investigation.

Critical review on emerging health effects associated with the indoor air quality and its sustainable management.

Indoor air quality (IAQ) is one of the fundamental elements affecting people's health and well-being. Currently, there is a lack of awareness among people about the quantification, identification, and possible health effects of IAQ. Airborne pollutants such as volatile organic compounds (VOCs), particulate matter (PM), sulfur dioxide (SO2), carbon monoxide (CO), nitrous oxide (NO), polycyclic aromatic hydrocarbons (PAHs) microbial spores, pollen, allergens, etc. primarily contribute to IAQ deterioration. This review discusses the sources of major indoor air pollutants, molecular toxicity mechanisms, and their effects on cardiovascular, ocular, neurological, women, and foetal health. Additionally, contemporary strategies and sustainable methods for regulating and reducing pollutant concentrations are emphasized, and current initiatives to address and enhance IAQ are explored, along with their unique advantages and potentials. Due to their longer exposure times and particular physical characteristics, women and children are more at risk for poor indoor air quality. By triggering many toxicity mechanisms, including oxidative stress, DNA methylation, epigenetic modifications, and gene activation, indoor air pollution can cause a range of health issues. Low birth weight, acute lower respiratory tract infections, Sick building syndromes (SBS), and early death are more prevalent in exposed residents. On the other hand, the main causes of incapacity and early mortality are lung cancer, chronic obstructive pulmonary disease, and cardiovascular disorders. It's crucial to acknowledge anticipated research needs and implemented efficient interventions and policies to lower health hazards.

The potential of 4D's approach in curbing antimicrobial resistance among bacterial pathogens.

INTRODUCTION: Antibiotics are life-saving drugs but irrational/inappropriate use leads to the emergence of antibiotic-resistant bacterial superbugs, making their treatment extremely challenging. Increasing antimicrobial resistance (AMR) among bacterial pathogens is becoming a serious public health concern globally. If ignorance persists, there would not be any antibiotics available to treat even a common bacterial infection in future. AREA COVERED: This article intends to collate and discuss the potential of 4D's (right Drug, Dose, Duration, and De-escalation of therapy) approach to tackle the emerging problem of AMR. For this, we searched PubMed, Google Scholar, Medline, and clinicaltrials.gov databases primarily using keywords 'optimal antibiotic therapy,' 'antimicrobial resistance,' 'higher versus lower dose antibiotic treatment,' 'shorter versus longer duration antibiotic treatment,' 'de-escalation study', and 'antimicrobial stewardship measures' and based on the findings, form and expressed our opinion. EXPERT OPINION: More efforts are needed for developing diagnostics for rapid, accurate, point-of-care, and cost-effective pathogen identification and antimicrobial susceptibility testing (AST) to facilitate rational use of antibiotics. Current dosing and duration of therapies also need to be redefined to maximize their impact. Furthermore, de-escalation approaches should be developed and encouraged in the clinic. This altogether will minimize selection pressure on the pathogens and reduce emergence of AMR.

Current status of pesticide effects on environment, human health and it's eco-friendly management as bioremediation: A comprehensive review.

Pesticides are either natural or chemically synthesized compounds that are used to control a variety of pests. These chemical compounds are used in a variety of sectors like food, forestry, agriculture and aquaculture. Pesticides shows their toxicity into the living systems. The World Health Organization (WHO) categorizes them based on their detrimental effects, emphasizing the relevance of public health. The usage can be minimized to a least level by using them sparingly with a complete grasp of their categorization, which is beneficial to both human health and the environment. In this review, we have discussed pesticides with respect to their global scenarios, such as worldwide distribution and environmental impacts. Major literature focused on potential uses of pesticides, classification according to their properties and toxicity and their adverse effect on natural system (soil and aquatic), water, plants (growth, metabolism, genotypic and phenotypic changes and impact on plants defense system), human health (genetic alteration, cancer, allergies, and asthma), and preserve food products. We have also described eco-friendly management strategies for pesticides as a green solution, including bacterial degradation, myco-remediation, phytoremediation, and microalgae-based bioremediation. The microbes, using catabolic enzymes for degradation of pesticides and clean-up from the environment. This review shows the importance of finding potent microbes, novel genes, and biotechnological applications for pesticide waste management to create a sustainable environment.

Identification of metabolite extraction method for targeted exploration of antimicrobial resistance associated metabolites of Klebsiella pneumoniae.

Antimicrobial resistant Klebsiella pneumoniae (K. pneumoniae), as being a pathogen of critical clinical concern, urgently demands effective therapeutic options. However, the discovery of novel antibiotics over the last three decades has declined drastically and necessitates exploring novel strategies. Metabolomic modulation has been the promising approach for the development of effective therapeutics to deal with AMR; however, only limited efforts have been made to-date, possibly due to the unavailability of suitable metabolites extraction protocols. Therefore, in order to establish a detailed metabolome of K. pneumoniae and identify a method for targeted exploration of metabolites that are involved in the regulation of AMR associated processes, metabolites were extracted using multiple methods of metabolites extraction (freeze-thaw cycle (FTC) and sonication cycle (SC) method alone or in combination (FTC followed by SC; FTC + SC)) from K. pneumoniae cells and then identified using an orbitrap mass analyzer (ESI-LC-MS/MS). A total of 151 metabolites were identified by using FTC, 132 metabolites by using FTC+SC, 103 metabolites by using SC and 69 metabolites common among all the methods used which altogether enabled the identification of 199 unique metabolites. Of these 199, 70 metabolites were known to have an association with AMR phenotype and among these, the FTC + SC method yielded better (identified 55 metabolites), quantitatively and qualitatively compared to FTC and SC alone (identified 51 and 41 metabolites respectively). Each method of metabolite extraction showed a definite degree of biasness and specificity towards chemical classes of metabolites and jointly contributed to the development of a detailed metabolome of the pathogen. FTC method was observed to give higher metabolomic coverage as compared to SC alone and FTC + SC. However, FTC + SC resulted in the identification of a higher number of AMR associated metabolites of K. pneumoniae compared to FTC and SC alone.