Competition of Cd(II) and Pb(II) on the bacterial cells: a new insight from bioaccumulation based on NanoSIMS imaging.

Polymetallic exposure causes complex toxicity to microorganisms. In this study, we investigated the responses of Escherichia coli under co-existence of cadmium (Cd) and lead (Pb), primarily based on biochemical analysis and RNA sequencing. Cd completely inhibited bacterial growth at a concentration of 2.41 mmol/L, with its removal rate as low as <10%. In contrast, the Pb removal rate was >95% under equimolar sole Pb stress. In addition, the Raman analysis confirmed the loss of proteins for the bacterial cells. Under the co-existence of Cd and Pb, the Cd toxicity to E. coli was alleviated. Meanwhile, the biosorption of Pb cations was more intense during the competitive sorption with Cd. Transmission electron microscopy images showed that a few cells were elongated during incubation, i.e., the average cellular length increased from 1.535 ± 0.407 to 1.845 ± 0.620 µm. Moreover, NanoSIMS imaging showed that the intracellular distribution of Cd and Pb was coupled with sulfur. Genes regulating sulfate transporter were also upregulated to promote sulfate assimilation. Then, the subsequent production of biogenic sulfide and sulfur-containing amino acids was enhanced. Although this strategy based on S enrichment could resist the polymetallic stress, not all related genes were induced to upregulate under sole Cd stress. Therefore, the S metabolism might remodel the microbial resistance to variable occurrence of heavy metals. Furthermore, the competitive sorption (in contrast to sole Cd stress) could prevent microbial cells from strong Cd toxicity.IMPORTANCEMicrobial tolerance and resistance to heavy metals have been widely studied under stress of single metals. However, the polymetallic exposure seems to prevail in the environment. Though microbial resistance can alleviate the effects of exogenous stress, the taxonomic or functional response to polymetallic exposure is still not fully understood. We determined the strong cytotoxicity of cadmium (Cd) on growth, and cell elongation would be driven by Cd stress. The addition of appropriate lead (Pb) showed a stimulating effect on microbial bioactivity. Meanwhile, the biosorption of Pb was more intense during co-existence of Pb and Cd. Our work also revealed the spatial coupling of intracellular S and Cd/Pb. In particular, the S assimilation was promoted by Pb stress. This work elucidated the microbial responses to polymetallic exposure and may provide new insights into the antagonistic function during metal stresses.

Extraction and application of extracellular polymeric substances from fungi.

Extracellular polymeric substances (EPS) are extracellular metabolites of microorganisms, highly associated with microbial function, adaptation, and growth. The main compounds in EPS have been revealed to be proteins, polysaccharides, nucleic acids, humic substances, lipids, etc. EPS are not only biomass, but also a biogenic material. EPS have high specific surface, abundant functional groups, and excellent degradability. In addition, they are more extensible to the environment than the microbial cells themselves, which exhibits their huge advantages. Therefore, they have been applied in many fields, such as the environment, ecosystem, basic commodities, and medicine. However, the functions of EPS highly depend on the suitable extraction process, as different extraction methods have different effects on their composition, structure, and function. There are many types of EPS extraction methods, in which physical and chemical methods have been widely utilized. This review summarizes the extraction methods and applications of EPS. In addition, it considers some important gaps in current knowledge, and indicates perspectives of EPS for their future study.

Transcriptome Analysis on Key Metabolic Pathways in Rhodotorula mucilaginosa Under Pb(II) Stress.

Rhodotorula mucilaginosa shows adaption to a broad range of Pb2+ stress. In this study, three key pathways, i.e., glycolysis (EMP), the tricarboxylic acid (TCA) cycle, and oxidative phosphorylation (OXPHOS), were investigated under 0-2,500 mg · L-1 Pb stress, primarily based on biochemical analysis and RNA sequencing. R. mucilaginosa cells showed similar metabolic response to low/medium (500/1000 mg · L-1) Pb2+ stress. High (2,500 mg · L-1) Pb2+ stress exerted severe cytotoxicity to R. mucilaginosa. The downregulation of HK under low-medium Pb2+ suggested a correlation with the low hexokinase enzymatic activity in vivo. However, IDH3, regulating a key step of circulation in TCA, was upregulated to promote ATP feedstock for downstream OXPHOS. Then, through activation of complex I & IV in the electron transport chain (ETC) and ATP synthase, ATP production was finally enhanced. This mechanism enabled fungal cells to compensate for ATP consumption under low-medium Pb2+ toxicity. Hence, R. mucilaginosa tolerance to such a broad range of Pb2+ concentrations can be attributed to energy adaption. In contrast, high Pb2+ stress caused ATP deficiency. Then, the subsequent degradation of intracellular defense systems further intensified Pb toxicity. This study correlated responses of EMP, TCA, and OXPHOS pathways in R. mucilaginosa under Pb stress, hence providing new insights into the fungal resistance to heavy metal stress. IMPORTANCE Glycolysis (EMP), the tricarboxylic acid (TCA) cycle, and oxidative phosphorylation (OXPHOS) are critical metabolism pathways for microorganisms to obtain energy during the resistance to heavy metal (HM) stress. However, these pathways at the genetic level have not been elucidated to evaluate their cytoprotective functions for Rhodotorula mucilaginosa under Pb stress. In this study, we investigated these three pathways based on biochemical analysis and RNA sequencing. Under low-medium (500-1,000 mg · L-1) Pb2+ stress, ATP production was stimulated mainly due to the upregulation of genes associated with the TCA cycle and the electron transport chain (ETC). Such an energy compensatory mechanism could allow R. mucilaginosa acclimation to a broad range of Pb2+ concentrations (up to 1000 mg · L-1). In contrast, high (2500 mg · L-1) Pb2+ stress exerted its excessive toxicity by provoking ATP deficiency and damage to intracellular resistance systems. This study provided new insights into R. mucilaginosa resistance to HM stress from the perspective of metabolism.

Association between food insecurity and kidney stones in the United States: Analysis of the National Health and Nutrition Examination Survey 2007-2014.

Purpose: Although food insecurity is a major public health concern associated with various diseases, the relationship between food insecurity and kidney stones remains unclear. We aimed to investigate the association between food insecurity and kidney stones in the US population. Materials and methods: Four continuous cycles of data from the year 2007 to 2014 were obtained from National Health and Nutrition Examination Survey (NHANES) in the current study. We utilized the Household Food Security Module to assess the food security status of individuals. The primary outcome was whether participants ever had kidney stones, and the secondary outcome was a stone recurrence. A multivariate logistic regression model adjusting for potential confounders was constructed to evaluate the independent association between food insecurity and kidney stones. Results: A total of 21,914 participants were included in our analysis, with 8.8% having a history of kidney stones and 33.2% of these reporting stones recurrence. Food insecurity was associated with increased risks of kidney stones (odds ratio: 1.21; 95% confidence interval: 1.05-1.39; P = 0.010) and kidney stones recurrence (odds ratio: 1.33; 95% confidence interval: 1.00-1.77; P = 0.052) after adjusting for all potential confounders. In addition, participants with very low food security had 38% and 47% higher risks of kidney stones (odds ratio: 1.38; 95% confidence interval: 1.13-1.69; P = 0.002; P for trend = 0.009) and kidney stones recurrence (odds ratio: 1.47; 95% confidence interval: 1.03-2.10; P = 0.032; P for trend = 0.029), respectively. Conclusion: There exists a significant association between food insecurity and kidney stones, which reveals the significance of the improvement of food insecurity in the alleviation of kidney stone formation and recurrence.

Histochemical analysis of the root tuber of Polygonum multiflorum Thunb. (Fam. Polygonaceae).

Authentication is the first priority in quality evaluation of Chinese herbal medicines (CHMs). The most commonly used authentication methods are morphological identification and microscopic identification. Unfortunately, these two methods cannot provide the chemical information needed to assess the quality of CHMs. In this study, a combination of fluorescence microscopy and high performance liquid chromatography-mass spectrometry (HPLC-MS) was used to analyze the chemical profiles of the tissues of the raw root tubers of Polygonum multiflorum Thunb. The results showed that the cork cells, cortex, and vessels in transverse sections of the raw root tuber of P. multiflorum fluoresced differently. Further analysis by HPLC-MS revealed that anthraquinones are mainly distributed in the cortex, and 2,3,5,4'-tetrahydroxystilbene-2-O-ß-D-glucopyranoside could be found in all tissues of the raw root tubers of P. multiflorum with its relative content as cork > cortex > xylem of allotype vascular bundles > xylem of central vascular bundles. Moreover, the fluorescence characteristics of the tissues from the steamed root tuber of P. multiflorum were compared and showed different fluorescence from those of raw material. From these results, it can be deduced that the root tuber of P. multiflorum with a broader cortex and fewer vascular bundles visible in a transverse section should be of better quality. The different fluorescence characteristics can be used to differentiate raw root tubers of P. multiflorum from those that have been steamed.