Metabolomic and proteomic responses of Phaeodactylum tricornutum to hypoxia

Diatoms are important contributors to global net primary productivity,and play a crucial role in the biogeochemical cycles of carbon,phosphorus,nitrogen,iron,and silicon.Currently in some regions in the ocean,there’s a trend that carbon content is high while oxygen concentration is low,and the underlying mechanisms of diatoms’response to low oxygen environments are worth investigating.Phaeodactylum tricornutum is a model diatom whose genome has been sequenced;it provides a universal molecular toolbox and a stable transgenic expression system.Therefore,the study of the responses of P.tricornutum to low oxygen has not only fundamental research significance but also important ecologic al significance.In this study,growth rates were determined and proteomic analysis and metabolomic analysis were performed to examine P.tricornutum responses under different oxygen concentrations(2%oxygen concentration for hypoxic condition and 21%oxygen concentration for the normal condition(sterilized air)).Results show that the hypoxic environment inhibited the growth of P.tricornutum.In the hypoxic conditions,P.tricornutum could reset its metabolism pathways,including enhancement in lipid utilization,replenishment of tricarboxylic acid(TCA)cycle through the glyoxylic acid cycle,and down-regulation of photorespiration to reduce energy waste.Additionally,the stress resistance mechanism was activated to facilitate the adaptation to low oxygen conditions.This study helps to reveal the different metabolic changes to hypoxia of diatom from that of higher plants,which might be ascribed to their different habitats and needs further exploration in the future.

Berberine reverses multidrug resistance in Candida albicans by hijacking the drug efflux pump Mdr1p

Clinical use of antimicrobials faces great challenges from the emergence of multidrug-resistant pathogens. The overexpression of drug efflux pumps is one of the major contributors to multidrug resistance(MDR). Reversing the function of drug efflux pumps is a promising approach to overcome MDR. In the life-threatening fungal pathogen Candida albicans, the major facilitator superfamily(MFS) transporter Mdr1p can excrete many structurally unrelated antifungals, leading to MDR. Here we report a counterintuitive case of reversing MDR in C. albicans by using a natural product berberine to hijack the overexpressed Mdr1p for its own importation. Moreover, we illustrate that the imported berberine accumulates in mitochondria and compromises the mitochondrial function by impairing mitochondrial membrane potential and mitochondrial Complex I. This results in the selective elimination of Mdr1 p overexpressed C. albicans cells. Furthermore, we show that berberine treatment can prolong the mean survival time of mice with blood-borne dissemination of Mdr1p overexpressed multidrug-resistant candidiasis. This study provides a potential direction of novel anti-MDR drug discovery by screening for multidrug efflux pump converters.

Integrating PCR-free amplification and synergistic sensing for ultrasensitive and rapid CRISPR/Cas12a-based SARS-CoV-2 antigen detection

Antigen detection provides particularly valuable information for medical diagnoses;however,the current detection methods are less sensitive and accurate than nucleic acid analysis.The combination of CRISPR/Cas12a and aptamers provides a new detection paradigm,but sensitive sensing and stable amplification in antigen detection remain challenging.Here,we present a PCR-free multiple trigger dsDNA tandem-based signal amplification strategy and a de novo designed dual aptamer synergistic sensing strategy.Integration of these two strategies endowed the CRISPR/Cas12a and aptamer-based method with ultra-sensitive,fast,and stable antigen detection.In a demonstration of this method,the limit of detection was at the single virus level(0.17 fM,approximately two copies/μL)in SARS-CoV-2 antigen nucleocapsid protein analysis of saliva or serum samples.The entire procedure required only 20 min.Given our system’s simplicity and modular setup,we believe that it could be adapted reasonably easily for general applications in CRISPR/Cas12a-aptamer-based detection.