Demographic patterns of two related desert shrubs with overlapping distributions in response to past climate changes.

Numerous studies have revealed that past geological events and climatic fluctuations had profoundly affected the genetic structure and demographic patterns of species. However, related species with overlapping ranges may have responded to such environmental changes in different ways. In this study, we compared the genetic structure and population dynamics of two typical desert shrubs with overlapping distributions in northern China, Nitraria tangutorum and Nitraria sphaerocarpa, based on chloroplast DNA (cpDNA) variations and species distribution models. We sequenced two cpDNA fragments (trnH-trnA and atpH-atpI) in 633 individuals sampled from 52 natural populations. Twenty-four chlorotypes, including eight rare chlorotypes, were identified, and a single dominant haplotype (H4) widely occurred in the entire geographical ranges of the two species. There were also a few distinctive chlorotypes fixed in different geographical regions. Population structure analyses suggested that the two species had significantly different levels of total genetic diversity and interpopulation differentiation, which was highly likely correlated with the special habitat preferences of the two species. A clear phylogeographic structure was identified to exist among populations of N. sphaerocarpa, but not exist for N. tangutorum. The neutral tests, together with the distribution of pairwise differences revealed that N. tangutorum experienced a sudden demographic expansion, and its expansion approximately occurred between 21 and 7 Kya before present, while a rapid range expansion was not identified for N. sphaerocarpa. The ecological niche modeling (ENM) analysis indicated that the potential ranges of two species apparently fluctuated during the past and present periods, with obvious contraction in the Last Glacial Maximum (LGM) and recolonization in the present, respectively, comparing to the Last Interglacial (LIG). These findings suggest that the two species extensively occurred in the Northwest of China before the Quaternary, and the current populations of them originated from a few separated glacial refugia following their habitat fragmentation in the Quarternary. Our results provide new insights on the impact of past geological and climatic fluctuations on the population dynamics of desert plants in northwestern China, and further enforce the hypothesis that there were several independent glacial refugia for these species during the Quaternary glaciations.

Reduction of selenite and tellurite by a highly metal-tolerant marine bacterium / Reducción de selenita y telurito por una bacteria marina altamente tolerante a los metales

Selenium (Se) and tellurium (Te) contaminations in soils and water bodies have been widely reported in recent years. Se(IV) and Te(IV) were regarded as their most dangerous forms. Microbial treatments of Se(IV)- and Te(IV)-containing wastes are promising approaches because of their environmentally friendly and sustainable advantages. However, the salt-tolerant microbial resources that can be used for selenium/tellurium pollution control are still limited since industrial wastewaters usually contain a large number of salts. In this study, a marine Shewanella sp. FDA-1 (FDA-1) was reported for efficient Se(IV) and Te(IV) reduction under saline conditions. Process and product analyses were performed to investigate the bioreduction processes of Se(IV) and Te(IV). The results showed that FDA-1 can effectively reduce Se(IV) and Te(IV) to Se0 and Te0 Se(IV)/Te(IV) to Se0/Te0 in 72 h, which were further confirmed by XRD and XPS analyses. In addition, enzymatic and RT‒qPCR assays showed that flavin-related proteins, reductases, dehydrogenases, etc., could be involved in the bioreduction of Se(IV)/Te(IV). Overall, our results demonstrate the ability of FDA-1 to reduce high concentrations of Se(IV)/or Te(IV) to Se0/or Te0 under saline conditions and thus provide efficient microbial candidate for controlling Se and Te pollution.(AU)

Deciphering variations in the surficial bacterial compositions and functional profiles in the intersection between North and South Yellow Sea.

The coastal ocean systems play paramount role in the nutrient biogeochemistry because of its interconnected environment. To gain a novel insight into coupling relationships between bacterial community, functioning properties and nutrient metabolism, we conducted analysis on the patterns and driving factors of planktonic bacterial functional community across subsurface water of marine ranching near the Yellow Sea in both summer and winter. Illumina HiSeq Sequencing and a corresponding set of biogeochemical data were used to assess distribution patterns of taxa, adaptive mechanism and metabolic function. Results demonstrated that Proteobacteria, Cyanobacteria, Actinobacteriota and Bacteroidota were dominant phyla both in summer and winter. Taxonomic profiles related to nutrient variation were found to be highly correlated with Dissolved Oxygen (DO) and Chlorophyll fluorescence (FLUO), and distinct diversity differences were also found between summer and winter samples. Functional activity in summer associated with the relative abundance of phototrophy and photoautotrophy were the highest in the subsurface water, while in winter the dominant functional properties were mainly include chemoheterotrophy and aerobic_ chemoheterotrophy. A significant difference related to functional activity between summer and winter, mainly representing ligninolysis and iron_respiration. In general, our study provides a framework for understanding the relative importance of environmental factors, temperature variation and nutrient availability in shaping the metabolic processes of aquatic microorganisms, particularly in ocean mariculture systems.

Reduction of selenite and tellurite by a highly metal-tolerant marine bacterium.

Selenium (Se) and tellurium (Te) contaminations in soils and water bodies have been widely reported in recent years. Se(IV) and Te(IV) were regarded as their most dangerous forms. Microbial treatments of Se(IV)- and Te(IV)-containing wastes are promising approaches because of their environmentally friendly and sustainable advantages. However, the salt-tolerant microbial resources that can be used for selenium/tellurium pollution control are still limited since industrial wastewaters usually contain a large number of salts. In this study, a marine Shewanella sp. FDA-1 (FDA-1) was reported for efficient Se(IV) and Te(IV) reduction under saline conditions. Process and product analyses were performed to investigate the bioreduction processes of Se(IV) and Te(IV). The results showed that FDA-1 can effectively reduce Se(IV) and Te(IV) to Se0 and Te0 Se(IV)/Te(IV) to Se0/Te0 in 72 h, which were further confirmed by XRD and XPS analyses. In addition, enzymatic and RT‒qPCR assays showed that flavin-related proteins, reductases, dehydrogenases, etc., could be involved in the bioreduction of Se(IV)/Te(IV). Overall, our results demonstrate the ability of FDA-1 to reduce high concentrations of Se(IV)/or Te(IV) to Se0/or Te0 under saline conditions and thus provide efficient microbial candidate for controlling Se and Te pollution.

Effects of suspended particles and dispersants on marine oil snow formation of crude oil/diesel oil.

Marine oil snow (MOS) potentially forms after an oil spill. To fully understand the mechanism of its formation, we investigated the effects of suspended particles (SP) and dispersants on MOS formation of crude oil and diesel oil by laboratory experiments. In the crude oil experiment, the SP concentration of 0.2 g L-1 was more suitable for crude oil MOS formation. The addition of dispersants significantly stimulated N and TV during MS/MOS formation of SP at 0.4 g L-1 and 0.8 g L-1 concentration (p < 0.05). Without SP, the dispersants also stimulated crude oil MOS formation. Furthermore, the concentration of SP had a significantly positive effect on the reduction of the total amount of N-alkanes (p < 0.05). In the diesel oil experiment, after adding dispersants to diesel oil, the maximum N, Dm, and TV values at a SP concentration of 0.2 g L-1 were significantly higher than those at 0.4 g L-1 and 0.8 g L-1 (p < 0.05). Besides, we found that dispersants stimulated MOS formation in diesel oil at a SP concentration of 0.2 g L-1. However, the dispersants had an inhibitory effect on diesel oil MOS formation without SP. Notably, the MOS formed by diesel oil appeared white, unlike the black MOS associated with crude oil. These findings are important for the environmental impact of oil spills and elevated SP concentrations.

Spatial-temporal variation of marine fishing activities responding to policy and social events in China.

The spatial-temporal variation of the impact of political and social factors such as (Fishing Moratorium, Marine Protected Areas, New Year, and COVID-19) on fishing activities offshore was evaluated based on fishing efforts data by AIS from 2013 to 2020 for China. It is found that the maximum fishing intensity occurs within 20-30 km of the coastline, the area within 40 km of the coastline covers 51% of fishing activities, and within 100 km of the coastline accounts for 92% of fishing activities. From 2013 to 2016, fishing hotspots showed a highly aggregated spread in the Bohai Sea and the East China Sea. Since 2017, China's hot fishing areas have gradually fragmented and southern moved. During the fishing moratorium, the fishing efforts decreased by 51% compared to the pre-moratorium, and after the moratorium, the fishing efforts increased by 81% compared to the pre-moratorium on average. We investigated fishing activities in and around 249 MPAs in China and found that 71% of MPAs are free of fishing, and average fishing intensity across MPAs is 35% lower than not protected areas. Regarding the social events, it is concluded that during the New Year holiday, the fishing efforts and fishing area were reduced by 79% and 73%, respectively, compared to the regular fishing period. The COVID-19 epidemic prompted a decrease in fishing efforts in 2020 for the first time since 2013, with the average fishing efforts in 2020 being 14% and 6% lower than in 2019 and 2017 to 2019, respectively.

How iron-bearing minerals affect the biological reduction of Sb(V): A newly discovered function of nitrate reductase.

As a toxic element of global concern, the elevated concentration of antimony (Sb) in the environment has attracted increasing attention. Microorganisms have been reported as important driving forces for Sb transformation. Iron (Fe) is the most important metal associated element of Sb, however, how Fe-bearing minerals affect the biological transformation of Sb is still unclear. In this study, the effects of Fe-bearing minerals on biological Sb(V) reduction were investigated by employing a marine Shewanella sp. CNZ-1 (CNZ-1). Our results showed that the presence of hematite, magnetite and ferrihydrite (1 g/L) resulted in a decrease in Sb(III) concentration of ~19-31 % compared to the Fe(III)-minerals free system. The calculated Sb(V) reduction rates are 0.0256 (R2 0.71), 0.0389 (R2 0.87), 0.0299 (R2 0.96) and 0.0428 (R2 0.95) h-1 in the hematite-, magnetite-, ferrihydrite-supplemented and Fe(III)-minerals free systems, respectively. The cube-shaped Sb2O3 was characterized as a reductive product by using XRD, XPS, FTIR, TG and SEM approaches. Differential proteomic analysis showed that flagellar protein, cytochrome c, electron transfer flavoprotein, nitrate reductase and polysulfide reductase (up-regulation >1.5-fold, p value <0.05) were supposed to be included in the electron transport pathway of Sb(V) reduction by strain CNZ-1, and the key role of nitrate reductases was further highlighted during this reaction process based on the RT-qPCR and confirmatory experiments. Overall, these findings are beneficial to understand the environmental fate of Sb in the presence of Fe-bearing minerals and provide guidance in developing the bacteria/enzyme-mediated control strategy for Sb pollution.

Identification and expression analysis of the small auxin-up RNA (SAUR) gene family in Lycium ruthenicum.

The plant hormone auxin regulates numerous aspects of plant growth and development, and small auxin-up RNA (SAUR) is the largest family of early auxin response genes in higher plants. SAUR has been implicated in the regulation of multiple biological processes. However, no comprehensive analysis of SAUR genes has been reported in Lycium ruthenicum. L. ruthenicum is a thorny shrub with very pronounced salt and drought tolerance, and studies have shown that stem thorns are related to drought tolerance in L. ruthenicum. In this study, the identification, phylogenetic analysis, and conserved motif prediction of SAUR genes were extensively explored. Furthermore, the tissue expression patterns of selected SAUR genes were assayed with quantitative real-time polymerase chain reaction (RT-qPCR). A total of 33 putative LrSAURs were identified and divided into three clusters in a phylogenetic tree of L. ruthenicum. MEME analysis identified 10 motifs in L. ruthenicum, and the results suggested that motif 1 and motif 3 were widely distributed. Analyzing the transcriptome data of stem thorns at four developmental stages indicated that LrSAURs were differentially expressed in L. ruthenicum, and could be divided into six expression patterns. The RT-qPCR analysis of 21 genes showed that LrSAUR2, LrSAUR8, LrSAUR9, LrSAUR11, LrSAUR12, and LrSAUR19 were mainly expressed in stems and stem thorns, and may be related to stem thorn development.

Multiple micronutrient deficiencies alter energy metabolism in host and gut microbiome in an early-life murine model.

Introduction: Micronutrients perform a wide range of physiological functions essential for growth and development. However, most people still need to meet the estimated average requirement worldwide. Globally, 2 billion people suffer from micronutrient deficiency, most of which are co-occurring deficiencies in children under age five. Despite decades of research, animal models studying multiple micronutrient deficiencies within the early-life period are lacking, which hinders our complete understanding of the long-term health implications and may contribute to the inefficacy of some nutritional interventions. Evidence supporting the Developmental Origins of Health and Disease (DOHaD) theory demonstrates that early-life nutritional deficiencies carry life-long consequences mediated through various mechanisms such as abnormal metabolic programming, stunting, altered body composition, and the gut microbiome. However, this is largely unexplored in the multiple micronutrient deficient host. Methods: we developed a preclinical model to examine undernutrition's metabolic and functional impact on the host and gut microbiome early in life. Three-week-old weanling C57BL/6N male mice were fed a low-micronutrient diet deficient in zinc, folate, iron, vitamin A, and vitamin B12 or a control diet for 4-weeks. Results: Our results showed that early-life multiple micronutrient deficiencies induced stunting, altered body composition, impaired glucose and insulin tolerance, and altered the levels of other micronutrients not depleted in the diet within the host. In addition, functional metagenomics profiling and a carbohydrate fermentation assay showed an increased microbial preference for simple sugars rather than complex ones, suggestive of a less developed microbiome in the low-micronutrient-fed mice. Moreover, we found that a zinc-only deficient diet was not sufficient to induce these phenotypes, further supporting the importance of studying co-occurring deficiencies. Discussion: Together, these findings highlight a previously unappreciated role of early-life multiple micronutrient deficiencies in shaping the metabolic phenome of the host and gut microbiome through altered glucose energy metabolism, which may have implications for metabolic disease later in life in micronutrient-deficient survivors.

High dimethylsulfoniopropionate concentrations in the surface seawater over the marginal seas of China and the Northwest Pacific Ocean during May-July of 2021 following La Niña.

The total dimethylsulfoniopropionate (DMSPt) concentrations over the surface seawater of China's marginal seas and the northwest Pacific Ocean (NWPO) in May-July 2021 (during the recessional period of La Niña) were analysed. The results showed that the DMSPt concentrations in the marginal seas of China varied from 4.73 to 775.96 nmol L-1, with an average value of 111.42 ± 129.30 nmol L-1 (average ± standard deviation). It was 2-12 times higher than those previously measured in the same seas and in the NWPO in this study. Significant positive correlations between DMSPt, chlorophyll-a and surface seawater temperature (SST) were observed in the SYS, the ECS and the NWPO. Moreover, their abnormally high SST was related to La Niña. These results suggested that high phytoplankton abundance was caused by abnormally high SST following La Niña, which further promoted DMSPt concentration increases. However, the increase of DMSPt was also related to other factors such as nutrients.

Sex differences in islet stress responses support female ß cell resilience.

OBJECTIVE: Pancreatic ß cells play a key role in maintaining glucose homeostasis; dysfunction of this critical cell type causes type 2 diabetes (T2D). Emerging evidence points to sex differences in ß cells, but few studies have examined male-female differences in ß cell stress responses and resilience across multiple contexts, including diabetes. Here, we address the need for high-quality information on sex differences in ß cell and islet gene expression and function using both human and rodent samples. METHODS: In humans, we compared ß cell gene expression and insulin secretion in donors with T2D to non-diabetic donors in both males and females. In mice, we generated a well-powered islet RNAseq dataset from 20-week-old male and female siblings with similar insulin sensitivity. Our unbiased gene expression analysis pointed to a sex difference in the endoplasmic reticulum (ER) stress response. Based on this analysis, we hypothesized female islets would be more resilient to ER stress than male islets. To test this, we subjected islets isolated from age-matched male and female mice to thapsigargin treatment and monitored protein synthesis, cell death, and ß cell insulin production and secretion. Transcriptomic and proteomic analyses were used to characterize sex differences in islet responses to ER stress. RESULTS: Our single-cell analysis of human ß cells revealed sex-specific changes to gene expression and function in T2D, correlating with more robust insulin secretion in human islets isolated from female donors with T2D compared to male donors with T2D. In mice, RNA sequencing revealed differential enrichment of unfolded protein response pathway-associated genes, where female islets showed higher expression of genes linked with protein synthesis, folding, and processing. This differential expression was physiologically significant, as islets isolated from female mice were more resilient to ER stress induction with thapsigargin. Specifically, female islets showed a greater ability to maintain glucose-stimulated insulin production and secretion during ER stress compared with males. CONCLUSIONS: Our data demonstrate sex differences in ß cell gene expression in both humans and mice, and that female ß cells show a greater ability to maintain glucose-stimulated insulin secretion across multiple physiological and pathological contexts.

Analysis of a genetic region affecting mouse body weight.

Genetic factors affect an individual's risk of developing obesity, but in most cases each genetic variant has a small effect. Discovery of genes that regulate obesity may provide clues about its underlying biological processes and point to new ways the disease can be treated. Preclinical animal models facilitate genetic discovery in obesity because environmental factors can be better controlled compared with the human population. We studied inbred mouse strains to identify novel genes affecting obesity and glucose metabolism. BTBR T+ Itpr3tf/J (BTBR) mice are fatter and more glucose intolerant than C57BL/6J (B6) mice. Prior genetic studies of these strains identified an obesity locus on chromosome 2. Using congenic mice, we found that obesity was affected by a ∼316 kb region, with only two known genes, pyruvate dehydrogenase kinase 1 (Pdk1) and integrin α 6 (Itga6). Both genes had mutations affecting their amino acid sequence and reducing mRNA levels. Both genes have known functions that could modulate obesity, lipid metabolism, insulin secretion, and/or glucose homeostasis. We hypothesized that genetic variation in or near Pdk1 or Itga6 causing reduced Pdk1 and Itga6 expression would promote obesity and impaired glucose tolerance. We used knockout mice lacking Pdk1 or Itga6 fed an obesigenic diet to test this hypothesis. Under the conditions we studied, we were unable to detect an individual contribution of either Pdk1 or Itga6 to body weight. During our studies, with conditions outside our control, we were unable to reproduce some of our previous body weight data. However, we identified a previously unknown role for Pdk1 in cardiac cholesterol metabolism providing the basis for future investigations. The studies described in this paper highlight the importance and the challenge using physiological outcomes to study obesity genes in mice.

Porphyra haitanensis Polysaccharides Attenuates Blood Lipid via Gut-Liver Axis in Diet-Induced High-Fat Mesocricetus auratus through Multiple Integrated Omics.

SCOPE: Hyperlipidemia is currently a global public health problem severely affecting people's physical and mental health, as well as their quality of life. METHODS AND RESULTS: The present study is aimed at revealing the mechanism of Porphyra haitanensis polysaccharide (PHP) in decreasing blood lipids by acting through gut-liver axis in Mesocricetus auratus fed a high-fat diet. PHP significantly prevented increases in serum total cholesterol, triglycerides and low-density lipoprotein cholesterol, and alleviated damage to liver cells induced by a high-fat diet M. auratus, in a dose-dependent manner. PHP promotes proliferation of Muribaculaceae and Faecalibaculum, thereby enhancing the production of butyric acid both in the colon and liver, particularly high-dose PHP (HPHP). Low-dose PHP (LPHP) promotes the expression of phosphatidylcholine metabolites and fatty acid transport genes, and inhibits the expression of genes involved in fat degradation (Abhd5), adipogenesis (Me1), fatty acid synthesis (Fasn and Pnpla3), and fatty acid chain elongation (Elovl6) in the liver. However, HPHP inhibits the expression of triglyceride metabolites and promotes the expression of fatty acid transporter (CD36), fatty acid oxidation (Acacb), and peroxisome proliferator-activated receptor gamma (PPARg) genes in the liver. CONCLUSION: PHP regulates lipid metabolism through the gut microbiota, and the gut-liver axis plays an important role in its hypolipidemic effects.

Exploring the variation of bacterial community and nitrogen transformation functional genes under the pressure of heavy metals in different coastal mariculture patterns.

Equilibrium in microbial dynamics and nitrogen transformation in the sediment is critical for maintaining healthy mariculture environment. However, our understanding about the impact of heavy metals on the bacterial community and nitrogen transformation functional genes in different mariculture patterns is still limited. Here, we analyzed 30 sediment samples in the vertical distribution from three different mariculture patterns mainly include open mariculture zone (K), closed mariculture pond (F) and pristine marine area (Q). Illumina MiSeq Sequencing was applied to investigate the bacterial community and structure in the sediment. Quantitative polymerase chain reaction (qPCR) was used to determine the effect of heavy metals on nitrogen transformation functional genes. Results showed that bacterial community and structure varied greatly in different mariculture patterns. Chloroflexi, Proteobacteria and Desulfobacterota were predominant phyla in the coastal mariculture area. High concentrations of heavy metals mainly enriched in the up layer (5-40 cm) of the sediment in the mariculture zone. The abundance of functional genes in the closed mariculture pond was much higher than the open mariculture zone and pristine marine area. And the high abundance of nitrification and denitrification functional genes mainly accumulated at the depth from 5 cm to 40 cm. Heavy metals content such as Fe, Cr, Mn, Ni, As, Cd, Pb and nutrient content NH4+-N, NO3--N and NO2--N were highly associated with bacterial community and nitrogen transformation functional genes. This study comprehensively elaborated the effect of heavy metals on the bacterial community and nitrogen transformation functional genes in different coastal mariculture patterns, indicating the possible role of closed mariculture pond in reducing nitrogen transformation efficiency, which will provide useful information for preventing pollution risk in the mariculture area.

Dynamic Ins2 Gene Activity Defines ß-Cell Maturity States.

Transcriptional and functional cellular specialization has been described for insulin-secreting ß-cells of the endocrine pancreas. However, it is not clear whether ß-cell heterogeneity is stable or reflects dynamic cellular states. We investigated the temporal kinetics of endogenous insulin gene activity using live cell imaging, with complementary experiments using FACS and single-cell RNA sequencing, in ß-cells from Ins2GFP knockin mice. In vivo staining and FACS analysis of islets from Ins2GFP mice confirmed that at a given moment, ∼25% of ß-cells exhibited significantly higher activity at the evolutionarily conserved insulin gene, Ins2. Live cell imaging over days captured Ins2 gene activity dynamics in single ß-cells. Autocorrelation analysis revealed a subset of oscillating cells, with mean oscillation periods of 17 h. Increased glucose concentrations stimulated more cells to oscillate and resulted in higher average Ins2 gene activity per cell. Single-cell RNA sequencing showed that Ins2(GFP)HIGH ß-cells were enriched for markers of ß-cell maturity. Ins2(GFP)HIGH ß-cells were also significantly less viable at all glucose concentrations and in the context of endoplasmic reticulum stress. Collectively, our results demonstrate that the heterogeneity of insulin production, observed in mouse and human ß-cells, can be accounted for by dynamic states of insulin gene activity.

Invasion of Spartina alterniflora on Zostera japonica enhances the abundances of bacteria by absolute quantification sequencing analysis.

Plant invasion can alter soil organic matter composition and indirectly impact estuary ecology; therefore, it is paramount to understand how plant invasion influences the bacterial community. Here, we present an absolute quantification 16S rRNA gene sequencing to investigate the bacterial communities that were collected from Zostera japonica and Spartina alterniflora covered areas and Z. japonica degradation areas in the Yellow River Estuary. Our data revealed that the absolute quantity of bacteria in the surface layer was significantly (p < .05) higher than that in the bottom and degradation areas. Following the invasion of S. alterniflora, the abundances of Bacteroidia, Acidimicrobiaceae, and Dehalococcoidaceaewere enriched in the S. alterniflora sediment. In addition, variations in the composition of sediment bacterial communities at the phylum level were the most intimately related to total organic carbon (TOC), and the content of heavy metals could reduce the abundance of bacteria. This study provided some information to understand the effects of S. alterniflora invasion on Z. japonica from the perspective of microbiome level.

Beta-cell specific Insr deletion promotes insulin hypersecretion and improves glucose tolerance prior to global insulin resistance.

Insulin receptor (Insr) protein is present at higher levels in pancreatic ß-cells than in most other tissues, but the consequences of ß-cell insulin resistance remain enigmatic. Here, we use an Ins1cre knock-in allele to delete Insr specifically in ß-cells of both female and male mice. We compare experimental mice to Ins1cre-containing littermate controls at multiple ages and on multiple diets. RNA-seq of purified recombined ß-cells reveals transcriptomic consequences of Insr loss, which differ between female and male mice. Action potential and calcium oscillation frequencies are increased in Insr knockout ß-cells from female, but not male mice, whereas only male ßInsrKO islets have reduced ATP-coupled oxygen consumption rate and reduced expression of genes involved in ATP synthesis. Female ßInsrKO and ßInsrHET mice exhibit elevated insulin release in ex vivo perifusion experiments, during hyperglycemic clamps, and following i.p. glucose challenge. Deletion of Insr does not alter ß-cell area up to 9 months of age, nor does it impair hyperglycemia-induced proliferation. Based on our data, we adapt a mathematical model to include ß-cell insulin resistance, which predicts that ß-cell Insr knockout improves glucose tolerance depending on the degree of whole-body insulin resistance. Indeed, glucose tolerance is significantly improved in female ßInsrKO and ßInsrHET mice compared to controls at 9, 21 and 39 weeks, and also in insulin-sensitive 4-week old males. We observe no improved glucose tolerance in older male mice or in high fat diet-fed mice, corroborating the prediction that global insulin resistance obscures the effects of ß-cell specific insulin resistance. The propensity for hyperinsulinemia is associated with mildly reduced fasting glucose and increased body weight. We further validate our main in vivo findings using an Ins1-CreERT transgenic line and find that male mice have improved glucose tolerance 4 weeks after tamoxifen-mediated Insr deletion. Collectively, our data show that ß-cell insulin resistance in the form of reduced ß-cell Insr contributes to hyperinsulinemia in the context of glucose stimulation, thereby improving glucose homeostasis in otherwise insulin sensitive sex, dietary and age contexts.

Effects of hyperinsulinemia on pancreatic cancer development and the immune microenvironment revealed through single-cell transcriptomics.

BACKGROUND: Hyperinsulinemia is independently associated with increased risk and mortality of pancreatic cancer. We recently reported that genetically reduced insulin production resulted in ~ 50% suppression of pancreatic intraepithelial neoplasia (PanIN) precancerous lesions in mice. However, only female mice remained normoglycemic, and only the gene dosage of the rodent-specific Ins1 alleles was tested in our previous model. Moreover, we did not delve into the molecular and cellular mechanisms associated with modulating hyperinsulinemia. METHODS: We studied how reduced Ins2 gene dosage affects PanIN lesion development in both male and female Ptf1aCreER;KrasLSL-G12D mice lacking the rodent-specific Ins1 gene (Ins1-/-). We generated control mice having two alleles of the wild-type Ins2 gene (Ptf1aCreER;KrasLSL-G12D;Ins1-/-;Ins2+/+) and experimental mice having one allele of Ins2 gene (Ptf1aCreER;KrasLSL-G12D;Ins1-/-;Ins2+/-). We then performed thorough histopathological analyses and single-cell transcriptomics for both genotypes and sexes. RESULTS: High-fat diet-induced hyperinsulinemia was transiently or modestly reduced in female and male mice, respectively, with only one allele of Ins2. This occurred without dramatically affecting glucose tolerance. Genetic reduction of insulin production resulted in mice with a tendency for less PanIN and acinar-to-ductal metaplasia (ADM) lesions. Using single-cell transcriptomics, we found hyperinsulinemia affected multiple cell types in the pancreas, with the most statistically significant effects on local immune cell types that were highly represented in our sampled cell population. Specifically, hyperinsulinemia modulated pathways associated with protein translation, MAPK-ERK signaling, and PI3K-AKT signaling, which were changed in epithelial cells and subsets of immune cells. CONCLUSIONS: These data suggest a potential role for the immune microenvironment in hyperinsulinemia-driven PanIN development. Together with our previous work, we propose that mild suppression of insulin levels may be useful in preventing pancreatic cancer by acting on multiple cell types.

Microbial detoxification of 2,4,6-tribromophenol via a novel process with consecutive oxidative and hydrolytic debromination: Biochemical, genetic and evolutionary characterization.

As a typical brominated flame retardants (BFRs), 2,4,6-tribromophenol (TBP) has serious hazard to the environmental health and its environmental fate has attracted considerable attention. Dehalogenation reaction plays key role in microbial TBP degradation and detoxification. So far, several halophenols-degrading enzymes have been reported to transform their substrate by oxidative dehalogenation; however, the molecular and biochemistry characterization of microbial hydrolytic dehalogenation is limited. In this study, Cupriavidus sp. CNP-8 with high TBP degradation activity was found to degrade TBP via an obviously differnet pathway as compared to other reported TBP-degraders. The transcription of hnp genes were significantly upregulated with TBP stimulation, indicating their involvment in TBP degradation. Enzymatic assays with 18O-labeling experiments showed that HnpAB, a two-component FAD-dependent monooxygenase, transformed TBP via consecutive oxidative and hydrolytic debromination reactions with the formation of 6-bromo-1,2,4-benzenetriol (BBT) as the ring-cleavage substrate. The function of the BBT ring-cleavage enzyme (HnpC) was also characterized both in vitro and in vivo. This finding provides new molecular mechanism of microbial detoxification of TBP and novel information of the environmental fate of this BFRs. Furthermore, to investigate the frequency of this novel dehalogenation mechanism in microbes, we also analyzed the distribution as well as the genetic structure of the hnpABC cluster by comparative genomics. Although hnpA homolog is distributed in several bacterial genera including Cupriavidus, Paraburkholderia, Variovorax and Streptomyces, the complete hnpABC cluster is only retrieved from Cupriavidus and strictly conservative in the genomes. This indicated that Cupriavidus have unique evolutionary pattern in acquiring the hnpABC to degrade TBP and its analogs, enhancing our understanding of the microbial adaptive evolution in halophenols-contaminated environment.