Fission and Internal Fusion of Protocell with Membraneless "Organelles" Formed by Liquid-Liquid Phase Separation.

Construction of protocells with hierarchical structures and living functions is still a great challenge. Growing evidence demonstrates that the membraneless organelles, which facilitate many essential cellular processes, are formed by RNA, protein, and other biopolymers via liquid-liquid phase separation (LLPS). The formation of the protocell in the early days of Earth could follow the same principle. In this work, we develop a novel coacervate-based protocell containing membraneless subcompartments via spontaneous liquid-liquid phase separation by simply mixing single-stranded oligonucleotides (ss-oligo), quaternized dextran (Q-dextran), and poly(l-lysine) (PLL) together. The resulting biphasic droplet, with PLL/ss-oligo phase being the internal subcompartments and Q-dextran/ss-oligo phase as the surrounding medium, is capable of sequestering and partitioning biomolecules into distinct regions. When the droplet is exposed to salt or Dextranase, the surrounding Q-dextran/ss-oligo phase will be gradually dissociated, resulting in the chaotic movement and fusion of internal subcompartments. Besides, the external electric field at a lower strength can drive the biphasic droplet to undergo a deviated circulation concomitant with the fusion of localized subcompartments, while a high-strength electric field can polarize the whole droplet, resulting in the release of daughter droplets in a controllable manner. Our study highlights that liquid-liquid phase separation of biopolymers is a powerful strategy to construct hierarchically structured protocells resembling the morphology and functions of living cells and provides a step toward a better understanding of the transition mechanism from nonliving to living matter under prebiotic conditions.

PEGylated gene carriers in serum under shear flow.

The behaviour of drug/gene carriers in the blood stream under shear is still a puzzle. In this work, using the complexes formed by 21 bp DNA and poly(ethylene glycol)-b-poly(l-lysine) (PEG-PLL) of varying PEG lengths, we studied the dynamic behaviour of the complexes in the presence of fetal bovine serum (FBS) and under flow at different shear rates, a condition mimicking the internal physical environment of blood vessels. The PEG5k-PLL/DNA complex possesses a dense DNA/PLL core and a loose PEG5k protecting layer. The PEGylated DNA complexes exhibit multiple responses to external shear in the presence of FBS. The loose PEG5k layer is firstly disturbed at a shear rate below 30 s-1. The exposure of the charged core to the environment results in a secondary aggregation of the complex with FBS. The size of the aggregate is limited to a certain range as the shear rate increases to 50 s-1. The dense DNA/PLL core starts to withstand the shear force as the shear rate reaches 500 s-1. The reorganization of the core to accommodate more serum molecules leads to tertiary aggregation of the complexes. If PEG cannot form a valid layer around the complex, as in PEG2k-PLL/DNA, the complex forms an aggregate even without shear, and the first shear dependent region is missing. If the PEG layer is too stable around the complex, as in PEG10k-PLL/DNA, no tertiary aggregation occurs. The mechanism of shear on the behaviour of delivery particles in serum helps to design gene carriers with high efficacy.

Estimating stomatal conductance of citrus under water stress based on multispectral imagery and machine learning methods.

Introduction: Canopy stomatal conductance (Sc) indicates the strength of photosynthesis and transpiration of plants. In addition, Sc is a physiological indicator that is widely employed to detect crop water stress. Unfortunately, existing methods for measuring canopy Sc are time-consuming, laborious, and poorly representative. Methods: To solve these problems, in this study, we combined multispectral vegetation index (VI) and texture features to predict the Sc values and used citrus trees in the fruit growth period as the research object. To achieve this, VI and texture feature data of the experimental area were obtained using a multispectral camera. The H (Hue), S (Saturation) and V (Value) segmentation algorithm and the determined threshold of VI were used to obtain the canopy area images, and the accuracy of the extraction results was evaluated. Subsequently, the gray level co-occurrence matrix (GLCM) was used to calculate the eight texture features of the image, and then the full subset filter was used to obtain the sensitive image texture features and VI. Support vector regression, random forest regression, and k-nearest neighbor regression (KNR) Sc prediction models were constructed, which were based on single and combined variables. Results: The analysis revealed the following: 1) the accuracy of the HSV segmentation algorithm was the highest, achieving more than 80%. The accuracy of the VI threshold algorithm using excess green was approximately 80%, which achieved accurate segmentation. 2) The citrus tree photosynthetic parameters were all affected by different water supply treatments. The greater the degree of water stress, the lower the net photosynthetic rate (Pn), transpiration rate (Tr), and Sc of the leaves. 3) In the three Sc prediction models, The KNR model, which was constructed by combining image texture features and VI had the optimum prediction effect (training set: R2 = 0.91076, RMSE = 0.00070; validation set; R2 = 0.77937, RMSE = 0.00165). Compared with the KNR model, which was only based on VI or image texture features, the R2 of the validation set of the KNR model based on combined variables was improved respectively by 6.97% and 28.42%. Discussion: This study provides a reference for large-scale remote sensing monitoring of citrus Sc by multispectral technology. Moreover, it can be used to monitor the dynamic changes of Sc and provide a new technique for gaining a better understanding of the growth status and water stress of citrus crops.

Serum Vitamin D Concentrations, Time to Pregnancy, and Pregnancy Outcomes among Preconception Couples: A Cohort Study in Shanghai, China.

BACKGROUND: The role of vitamin D in reproductive health is still unclear. This study aimed to assess the effect of serum 25-hydroxyvitamin D (25(OH)D), among preconception couples, on fecundity, and the associations between 25(OH)D concentrations before and during pregnancy, and pregnancy outcomes. METHODS: 200 preconception couples attempting to conceive were recruited and were followed-up until childbirth. Time to pregnancy was collected via telephone every two months or obtained via a questionnaire during pregnancy. Blood samples were collected to measure serum 25(OH)D levels from both partners at enrollment and from women during the second and third trimester of pregnancy. RESULTS: Couples had higher conception rates within six months (adjusted odds ratio (aOR): 3.72, 95% CI: 1.16, 11.9) and reduced time to pregnancy (adjusted fecundability ratio (aFR): 1.50, 95% CI: 1.01, 2.23) if male partners had sufficient 25(OH)D compared with insufficient 25(OH)D. Compared to pregnant women with insufficient 25(OH)D in the third trimester of pregnancy, sufficient 25(OH)D was associated with reduced odds of anemia (OR: 0.22, 95% CI: 0.06, 0.82), longer gestational age (ß: 0.53, 95% CI: 0.04, 1.01) and newborns' higher ponderal index (ß: 0.10, 95% CI: 0.01, 0.19). CONCLUSIONS: Sufficient serum 25(OH)D levels among preconception men or during pregnancy were associated with better reproductive health.

Distributions and contamination assessment of heavy metals in the surface sediments of western Laizhou Bay: Implications for the sources and influencing factors.

Heavy metals (Cu, Pb, Cr, Cd and As) contents in surface sediments from western Laizhou Bay were analysed to evaluate the spatial distribution pattern and their contamination level. As was mainly concentrated in the coastal area near the estuaries and the other metals were mainly concentrated in the central part of the study area. The heavy metals were present at unpolluted levels overall evaluated by the sediment quality guidelines and geoaccumulation index. Principal component analysis suggest that Cu, Pb and Cd were mainly sourced from natural processes and As was mainly derived from anthropogenic inputs. Meanwhile, Cr originated from both natural processes and anthropogenic contributions. Tidal currents, sediments and human activities were important factors affecting the distribution of heavy metals. The heavy metal environment was divided into four subareas to provide a reference for understanding the distribution and pollution of heavy metals in the estuary-bay system.