Monocyte distribution width as an early predictor of short-term outcome in adult patients with sepsis.

OBJECTIVES: Monocyte distribution width (MDW) is a quantitative measurement of monocyte anisocytosis and has been proposed as an efficient marker for early sepsis detection. This study aimed to assess the prognostic potential of MDW in septic patients. METHODS: In this study, a total of 252 adult septic patients were enrolled. Demographic, clinical, and laboratory finding including MDW and traditional inflammatory biomarkers detected at three time points (day 1, day 3 and day 6) after admission were collected and compared between 28-day survivors and non-survivors. Receiver operating characteristic (ROC) curves, Kaplan-Meier survival curve and Cox regression analyses were performed to assess and compare their predictive values. Group-based trajectory modeling was applied to identify MDW trajectory endotypes. Basic characteristics and 28-day outcomes were compared between the trajectories. RESULTS: ROC curve analysis showed that MDW levels measured on day 3 after admission (D3-MDW) had moderate prognostic value and was independently associated with 28-day mortality in patients with sepsis. A D3-MDW value of 26.20 allowed discrimination between survivors and non-survivors with a sensitivity of 77.8 % and a specificity of 67.6 %. However, the prognostic accuracy of D3-MDW was diminished in immune-compromised patients and patients who already received antibiotics before admission. Group-based trajectory modeling indicated that excessively elevated and delayed decreased MDW levels during the first week after admission inversely correlated with prognosis. CONCLUSIONS: MDW values detected on day 3 after admission and its kinetic change might be potential markers for predicting short-term outcome in adult septic patients.

Antibiotics in urine from general adults in Shenzhen, China: Demographic-related difference in exposure levels.

Misuse or overuse of antibiotics can have a variety of detrimental microbial effects. However, the body burden of antibiotics in the general population is currently unclear. In this cross-sectional study, we determined four classes of widely-applied antibiotics (3 imidazoles, 2 sulfonamides, 5 quinolones, and 2 chloramphenicols) in urine samples from 1170 adult residents in Shenzhen, China. Antibiotics were detected in 30.8 % of all urine samples with concentrations ranging from

Arsenic speciation in shellfish from South China Sea: Levels, estimated daily intake and health risk assessment.

The purposes of this study were to measure the concentrations of arsenic speciation in shellfish from South China Sea and evaluate the health risk by local residents through shellfish consumption. The median concentrations (in wet weight) of arsenic speciation in shellfish samples were in the following order: AsB (16.0 mg·kg-1) > DMA (1.30 mg·kg-1) > AsV (0.23 mg·kg-1) > AsC (0.08 mg·kg-1) > AsIII (0.05 mg·kg-1) > MMA (0.01 mg·kg-1). Among shellfish species, Mactra mera and Babylonia areolata were found to accumulate iAs and AsB, respectively. The target hazard quotient values of iAs (THQiAs) in all shellfish samples were lower than 1. However, the carcinogenic risk values of iAs (CRiAs) in the Mactra mera, Mytilus galloprovincialis and Pinctada margaritifera were beyond the acceptable range, implying that continuous exposure to iAs pollution via the consumption of these shellfish would pose a potential cancer risk to local consumers.

Urinary parabens, bisphenol A and triclosan in primiparas from Shenzhen, China: Implications for exposure and health risks.

The usage of parabens, bisphenol A and triclosan in diverse consumer products is in widespread. Nevertheless, there are limited data concerning exposure to these chemicals in human being, especially in primiparas. Biomonitoring of chemicals in primiparas is useful for the estimation of chemical exposure risks for both primiparas and their offspring. This study aims to investigate urinary levels of parabens, bisphenol A and triclosan of 84 primiparas from Shenzhen, China and to evaluate their potential health risks. Methyl, ethyl, and n-propyl parabens bisphenol A and triclosan exhibited high detection rates (DRs) (> 97%) in urine samples, suggesting that primiparas are exposed to them widely. The median concentrations of methyl, ethyl, and n-propyl parabens, bispenol A and triclosan in urine were 2.14, 4.10, 0.46, 1.30 and 3.00 µg/L, respectively. Ethyl paraben was the predominant paraben accounting for nearly half of Σ3parabens (The sum concentrations of methyl, ethyl, n-propyl parabens). Positive associations with significance (p < 0.05) were found between the usage of plastic containers and urinary concentrations of ethyl paraben or BPA, indicating plastic containers might be an important factor influencing primipara exposure to these two chemicals. Urinary concentrations of methyl paraben were positively associated (p < 0.05) with the time of computer use by participant, suggesting that indoor dust might constitute an important source of parabens. The estimated daily intakes of parabens, bisphenol A and triclosan contrasted with the acceptable daily intakes in a comparatively low level. The hazard quotients (HQs) of these chemicals were all less than 1, suggesting no health risks for primiparas from South China. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s40201-020-00599-1.

GsSnRK1 interplays with transcription factor GsERF7 from wild soybean to regulate soybean stress resistance.

Although the function and regulation of SnRK1 have been studied in various plants, its molecular mechanisms in response to abiotic stresses are still elusive. In this work, we identified an AP2/ERF domain-containing protein (designated GsERF7) interacting with GsSnRK1 from a wild soybean cDNA library. GsERF7 gene expressed dominantly in wild soybean roots and was responsive to ethylene, salt, and alkaline. GsERF7 bound GCC cis-acting element and could be phosphorylated on S36 by GsSnRK1. GsERF7 phosphorylation facilitated its translocation from cytoplasm to nucleus and enhanced its transactivation activity. When coexpressed in the hairy roots of soybean seedlings, GsSnRK1(wt) and GsERF7(wt) promoted plants to generate higher tolerance to salt and alkaline stresses than their mutated species, suggesting that GsSnRK1 may function as a biochemical and genetic upstream kinase of GsERF7 to regulate plant adaptation to environmental stresses. Furthermore, the altered expression patterns of representative abiotic stress-responsive and hormone-synthetic genes were determined in transgenic soybean hairy roots after stress treatments. These results will aid our understanding of molecular mechanism of how SnRK1 kinase plays a cardinal role in regulating plant stress resistances through activating the biological functions of downstream factors.

Ectopic expression of a rice plasma membrane intrinsic protein (OsPIP1;3) promotes plant growth and water uptake.

Plasma membrane intrinsic proteins (PIPs) are known to be major facilitators of the movement of a number of substrates across cell membranes. From a drought-resistant cultivar of Oryza sativa (rice), we isolated an OsPIP1;3 gene single-nucleotide polymorphism (SNP) that is mostly expressed in rice roots and is strongly responsive to drought stress. Immunocytochemistry showed that OsPIP1;3 majorly accumulated on the proximal end of the endodermis and the cell surface around the xylem. Expression of GFP-OsPIP1;3 alone in Xenopus oocytes or rice protoplasts showed OsPIP1;3 mislocalization in the endoplasmic reticulum (ER)-like neighborhood, whereas co-expression of OsPIP2;2 recruited OsPIP1;3 to the plasma membrane and led to a significant enhancement of water permeability in oocytes. Moreover, reconstitution of 10×His-OsPIP1;3 in liposomes demonstrated water channel activity, as revealed by stopped-flow light scattering. Intriguingly, by patch-clamp technique, we detected significant NO3- conductance of OsPIP1;3 in mammalian cells. To investigate the physiological functions of OsPIP1;3, we ectopically expressed the OsPIP1;3 gene in Nicotiana benthamiana (tobacco). The transgenic tobacco plants exhibited higher photosynthesis rates, root hydraulic conductivity (Lpr ) and water-use efficiency, resulting in a greater biomass and a higher resistance to water deficit than the wild-type did. Further experiments suggested that heterologous expression of OsPIP1;3 in cyanobacterium altered bacterial growth under different conditions of CO2 gas supply. Overall, besides shedding light on the multiple functions played by OsPIP1;3, this work provides insights into the translational value of plant AQPs.

Identification of novel interactors and potential phosphorylation substrates of GsSnRK1 from wild soybean (Glycine soja).

The plant sucrose nonfermenting kinase 1 (SnRK1) kinases play the central roles in the processes of energy balance, hormone perception, stress resistance, metabolism, growth, and development. However, the functions of these kinases are still elusive. In this study, we used GsSnRK1 of wild soybean as bait to perform library-scale screens by the means of yeast two-hybrid to identify its interacting proteins. The putative interactions were verified by yeast retransformation and ß-galactosidase assays, and the selected interactions were further confirmed in planta by bimolecular fluorescence complementation and biochemical Co-IP assays. Protein phosphorylation analyses were carried out by phos-tag assay and anti-phospho-(Ser/Thr) substrate antibodies. Finally, we obtained 24 GsSnRK1 interactors and several putative substrates that can be categorized into SnRK1 regulatory ß subunit, protein modification, biotic and abiotic stress-related, hormone perception and signalling, gene expression regulation, water and nitrogen transport, metabolism, and unknown proteins. Intriguingly, we first discovered that GsSnRK1 interacted with and phosphorylated the components of soybean nodulation and symbiotic nitrogen fixation. The interactions and potential functions of GsSnRK1 and its associated proteins were extensively discussed and analysed. This work provides plausible clues to elucidate the novel functions of SnRK1 in response to variable environmental, metabolic, and physiological requirements.

Identification of Trypanosoma brucei AdoMetDC Inhibitors Using a High-Throughput Mass Spectrometry-Based Assay.

Human African trypanosomiasis (HAT) is a fatal infectious disease caused by the eukaryotic pathogen Trypanosoma brucei (Tb). Available treatments are difficult to administer and have significant safety issues. S-Adenosylmethionine decarboxylase (AdoMetDC) is an essential enzyme in the parasite polyamine biosynthetic pathway. Previous attempts to develop TbAdoMetDC inhibitors into anti-HAT therapies failed due to poor brain exposure. Here, we describe a large screening campaign of two small-molecule libraries (∼400,000 compounds) employing a new high-throughput (∼7 s per sample) mass spectrometry-based assay for AdoMetDC activity. As a result of primary screening, followed by hit confirmation and validation, we identified 13 new classes of reversible TbAdoMetDC inhibitors with low-micromolar potency (IC50) against both TbAdoMetDC and T. brucei parasite cells. The majority of these compounds were >10-fold selective against the human enzyme. Importantly, compounds from four classes demonstrated high propensity to cross the blood-brain barrier in a cell monolayer assay. Biochemical analysis demonstrated that compounds from eight classes inhibited intracellular TbAdoMetDC in the parasite, although evidence for a secondary off-target component was also present. The discovery of several new TbAdoMetDC inhibitor chemotypes provides new hits for lead optimization programs aimed to deliver a novel treatment for HAT.

Relief of autoinhibition by conformational switch explains enzyme activation by a catalytically dead paralog.

Catalytically inactive enzyme paralogs occur in many genomes. Some regulate their active counterparts but the structural principles of this regulation remain largely unknown. We report X-ray structures of Trypanosoma brucei S-adenosylmethionine decarboxylase alone and in functional complex with its catalytically dead paralogous partner, prozyme. We show monomeric TbAdoMetDC is inactive because of autoinhibition by its N-terminal sequence. Heterodimerization with prozyme displaces this sequence from the active site through a complex mechanism involving a cis-to-trans proline isomerization, reorganization of a ß-sheet, and insertion of the N-terminal α-helix into the heterodimer interface, leading to enzyme activation. We propose that the evolution of this intricate regulatory mechanism was facilitated by the acquisition of the dimerization domain, a single step that can in principle account for the divergence of regulatory schemes in the AdoMetDC enzyme family. These studies elucidate an allosteric mechanism in an enzyme and a plausible scheme by which such complex cooperativity evolved.

Water and CO2 permeability of SsAqpZ, the cyanobacterium Synechococcus sp. PCC7942 aquaporin.

BACKGROUND INFORMATION: Cyanobacteria possess Aquaporin-Z (AqpZ) membrane channels which have been suggested to mediate the water efflux underlying osmostress-inducible gene expression and to be essential for glucose metabolism under photomixotrophic growth. However, preliminary observations suggest that the biophy-sical properties of transport and physiological meaning of AqpZ in such photosynthetic microorganisms are not yet completely assessed. RESULTS: In this study, we used Xenopus laevis oocyte and proteoliposome systems to directly demonstrate the water permeability of the cyanobacterium Synechococcus sp. PCC7942 aquaporin, SsAqpZ. By an in vitro assay of intracellular acidification in yeast cells, SsAqpZ was found to transport also CO2 . Consistent with this result, during the entire exponential phase of growth, Synechococcus SsAqpZ-null-mutant cells grew slower than the corresponding wild-type cells. This phenotype was stronger with higher levels of extracellular CO2 . In line with the conversion of CO2 gas into HCO3(-) ions under alkaline conditions, the impairment in growth of the SsAqpZ-null strain was weaker in more alkaline culture medium. CONCLUSIONS: Cyanobacterial SsAqpZ may exert a pleiotropic function in addition to the already reported roles in macronutrient homeostasis and osmotic-stress response as it appears to constitute an important pathway in CO2 uptake, a fundamental step in photosynthesis.

Aquaporin OsPIP1;1 promotes rice salt resistance and seed germination.

OsPIP1;1 is one of the most abundant aquaporins in rice leaves and roots and is highly responsible to environmental stresses. However, its biochemical and physiological functions are still largely unknown. The oocyte assay data showed OsPIP1;1 had lower water channel activity in contrast to OsPIP2;1. EGFP and immunoelectron microscopy studies revealed OsPIP1;1 was predominantly localized in not only plasma membrane but also in some ER-like intracellular compartments in the cells. OsPIP1;1 exhibited low water channel activity in Xenopus oocytes but coexpression of OsPIP2;1 significantly enhanced its water permeability. Stop-flow assay indicated that 10His-OsPIP1;1-reconstituted proteoliposomes had significantly higher water permeability than the control liposomes. Overexpression of OsPIP1;1 greatly altered many physiological features of transgenic plants in a dosage-dependent manner. Moderate expression of OsPIP1;1 increased rice seed yield, salt resistance, root hydraulic conductivity, and seed germination rate. This work suggests OsPIP1;1 functions as an active water channel and plays important physiological roles.

2-Eth-oxy-ethyl (Z)-2-cyano-3-[(N-phenyl-carbamo-yl)amino]-prop-2-enoate.

The crystal structure of the title compound, C(15)H(17)N(3)O(4), is stabilized by inter-molecular N-H⋯N hydrogen bonds. An intra-molecular N-H⋯O hydrogen bond also occurs.

3-Benzyl-amino-2-cyano-N-[N-(2-fluoro-phenyl)-carbamo-yl]-3-(methyl-sulfanyl)acryl-amide.

In the crystal structure of the title compound, C(19)H(17)FN(4)O(2)S, mol-ecules are linked via pairs of N-H⋯N inter-actions, forming centrosymmetric dimers. Two intra-molecular N-H⋯O hydrogen bonds stabilize the mol-ecular conformation.