Influence of polyethylene glycol density and surface lipid on pharmacokinetics and biodistribution of lipid-calcium-phosphate nanoparticles

The pharmacokinetics (PK) and biodistribution of nanoparticles (NPs) are controlled by a complex array of interrelated, physicochemical and biological factors of NPs. The lipid-bilayer core structure of the Lipid-Calcium-Phosphate (LCP) NPs allows us to examine the effects of the density of polyethylene glycol (PEG) and the incorporation of various lipids onto the surface on their fate in vivo. Fluorescence quantification estimated that up to 20% (molar percent of outer leaflet lipids) could be grafted on the surface of LCP NPs. Contrary to the common belief that high level of PEGylation could prevent the uptake of NPs by the reticuloendothelial system (RES) organs such as liver and spleen, a significant amount of the injected dose was observed in the liver. Confocal microscopy revealed that LCP NPs were largely localized in hepatocytes not Kupffer cells. It was further demonstrated that the delivery to hepatocytes was dependent on both the concentration of PEG and the surface lipids. LCP NPs could be directed from hepatocytes to Kupffer cells by decreasing PEG concentration on the particle surface. In addition, LCP NPs with 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) exhibited higher accumulation in the hepatocytes than LCP NPs with dioleoylphosphatidylcholine (DOPC). Analysis of the proteins bound to NPs suggested that apolipoprotein E (apoE) might serve as an endogenous targeting ligand for LCP-DOTAP NPs, but not LCP-DOPC NPs. The significant uptake of NPs by the hepatocytes is of great interest to formulation design for oncologic and hepatic drug deliveries

Transcriptome analysis reveals the molecular mechanisms of rubber biosynthesis and laticifer differentiation during rubber seed germination

The molecular mechanisms underlying the initiation of natural rubber synthesis and laticifer differentiation have not been fully elucidated. In this study, we conducted a time-series transcriptome analysis of five rubber tree tissues at four stages of seed germination. A total of 161,199 DEGs were identified between the two groups, including most 16,673 DEGs (A3 vs B3 and A3 vs C3) and lest 1,210 DEGs (C2 vs D2). We found that the maturation of the seed is accompanied by the formation of laticifer cells in cotyledon. Meanwhile, the analysis of hormones related genes expression may provide effective clues for us to promote the differentiation of laticifer cells in seeds by hormones in the future. In this study, hormone-related gene enrichment analyses revealed that IAA, GA, and CTK were activated in laticifer containing tissues. Similarly, GO and GEGG analysis showed that hormone pathways, especially the auxin pathway, are enriched. Gene expression clustering was analyzed using the short time-series expression miner (STEM), and the analysis revealed four distinct trends in the gene expression profiles. Moreover, we enriched transcription factor (TF) enrichment in cotyledon and embryonic axis tissues, and the MYB type exhibited the most significant difference. Furthermore, our findings revealed that genes related to rubber synthesis exhibited tissue-specific expression patterns during seed germination. Notably, key genes associated with rubber biosynthesis, specifically small rubber particle protein (SRPP) and cis-prenyltransferase (CPT), exhibited significant changes in expression in cotyledon and embryonic axis tissues, suggesting synchronous rubber synthesis with seed germination. Our staining results reveled that laticifer cells were exits in the cotyledon before seed imbibition stage. In conclusion, these results lay the foundation for exploring the molecular mechanisms underlying laticifer differentiation and rubber synthesis during seed germination, deepening our understanding of the initiation stages of rubber biosynthesis and laticifer differentiation

Cyclic intensive light exposure induces retinal lesions similar to age-related macular degeneration in APPswe/PS1 bigenic mice

<p>Abstract</p> <p>Background</p> <p>Intensive light exposure and beta-amyloid (Aβ) aggregates have been known as a risk factor for macular degeneration and an important component in the pathologic drusen structure involved in this disorder, respectively. However, it is unknown whether Aβ deposition mediates or exacerbates light exposure-induced pathogenesis of macular degeneration. Several studies including the one from us already showed accumulation of Aβ deposits in the retina in Alzheimer's transgenic mice. Using histopathological analysis combined with electroretinographic functional assessment, we investigated the effects of cyclic intensive light exposure (CILE) on the architecture of retina and related function in the APPswe/PS1bigenic mouse.</p> <p>Results</p> <p>Histopathological analysis has found significant loss of outer nuclear layer/photoreceptor outer segment and outer plexiform layer along with abnormal hypo- and hyper-pigmentation in the retinal pigment epithelium (RPE), remarkable choroidal neovascularization (CNV), and exaggerated neuroinflammatory responses in the outer retina of APPswe/PS1 bigenic mice following cyclic intensive light exposure (CILE), whereas controls remained little change contrasted with age-matched non-transgenic littermates. CILE-induced degenerative changes in RPE are further confirmed by transmission electron microcopy and manifest as formation of basal laminar deposits, irregular thickening of Bruch's membrane (BrM), deposition of outer collagenous layer (OCL) in the subretinal space, and vacuolation in the RPE. Immunofluorescence microscopy reveals drusenoid Aβ deposits in RPE as well as neovessels attached which are associated with disruption of RPE integrity and provoked neuroinflammatory response as indicated by markedly increased retinal infiltration of microglia. Moreover, both immunohistochemistry and Western blots detect an induction of vascular endothelial growth factor (VEGF) in RPE, which corroborates increased CNV in the outer retina in the bigenic mice challenged by CILE.</p> <p>Conclusions</p> <p>Our findings demonstrate that degenerative changes in the outer retina in the APPswe/PS1 bigenic mouse induced by CILE are consistent with these in AMD. These results suggest that an Alzheimer's transgenic animal model with accumulation of Aβ deposits might be an alternative animal model for AMD, if combined with other confounding factors such as intensive light exposure for AMD.</p

The effect of maternal vitamin D deficiency during pregnancy on glycolipid metabolism of offspring rats and the improvement of vitamin D intervention after weaning

BackgroundVitamin D deficiency during pregnancy is common, but whether maternal vitamin D status affects glycolipid metabolism of offspring remains unclear.ObjectiveTo evaluate the effect of maternal vitamin D deficiency during pregnancy on the glycolipid metabolism of offspring at different life-cycles (from birth to adulthood) and to explore the improvement of different dosages of vitamin D supplementation.MethodsSprague–Dawley rats were fed vitamin D-deprived (VDD group) or standard vitamin D diets (SC group) during pregnancy, and their diets were changed to standard vitamin D diets during lactation (the offspring were sorted into VDDoffspring and SCoffspring groups). After weaning, rats in the VDDoffspring group were randomly assigned to the VDDoffspring, VDDoffspring-S3300 and VDDoffspring-S10000 groups with diets containing standard, medium and high dosages of vitamin D for 12 wk. Serum was collected for biochemical analyses at postnatal Day 21, postnatal Day 56 and postnatal Day 84. Oral glucose tolerance test (OGTT) was performed at postnatal Day 70.ResultsCompared to SCoffspring, rats in the VDDoffspring group had significantly lower birth weight with faster weight gain and higher levels of lipid metabolism in early life. After near adulthood, the differences in weight and lipid metabolism between the two groups disappeared. OGTT showed significantly higher blood glucose levels in the VDDoffspring group at 30 min, 60 min, and 90 min. The continuation of vitamin D supplementation at medium and high dosages after weaning did not cause any obvious changes in weight or glycolipid metabolism (except for postprandial hyperglycemia). OGTT demonstrated that the glucose levels in the VDDoffspring-S3300 group were lowest at all the time points and that those in the VDDoffspring-S10000 group were the highest at 30 min, 60 min, and 90 min among the three groups.ConclusionThe adverse effects of vitamin D deficiency during pregnancy on glycolipid metabolism in offspring vary in different stages. Over a long time period, adequate vitamin D supplementation is beneficial to glycolipid metabolism for the offspring of subjects with vitamin D deficiency during pregnancy; however, further improvement is required

Effects of EMG-based robot for upper extremity rehabilitation on post-stroke patients: a systematic review and meta-analysis

Objective: A growing body of research shows the promise and efficacy of EMG-based robot interventions in improving the motor function in stroke survivors. However, it is still controversial whether the effect of EMG-based robot is more effective than conventional therapies. This study focused on the effects of EMG-based robot on upper limb motor control, spasticity and activity limitation in stroke survivors compared with conventional rehabilitation techniques.Methods: We searched electronic databases for relevant randomized controlled trials. Outcomes included Fugl-Meyer assessment scale (FMA), Modified Ashworth Scale (MAS), and activity level.Result: Thirteen studies with 330 subjects were included. The results showed that the outcomes post intervention was significantly improved in the EMG-based robot group. Results from subgroup analyses further revealed that the efficacy of the treatment was better in patients in the subacute stage, those who received a total treatment time of less than 1000 min, and those who received EMG-based robotic therapy combined with electrical stimulation (ES).Conclusion: The effect of EMG-based robot is superior to conventional therapies in terms of improving upper extremity motor control, spasticity and activity limitation. Further research should explore optimal parameters of EMG-based robot therapy and its long-term effects on upper limb function in post-stroke patients.Systematic Review Registration:https://www.crd.york.ac.uk/PROSPERO/; Identifier: 387070

Effects of Dietary Cholesterol Levels on the Growth, Molt Performance, and Immunity of Juvenile Swimming Crab, Portunus trituberculatus

The effects of dietary cholesterol levels on growth, molt performance, and immunity of juvenile swimming crab Portunus trituberculatus, were investigated at four cholesterol levels (0.2%-1.4%) of purified diets. Each diet was fed in triplicate to 18 crabs per replicate for 50 days. Crabs fed the diet with 1.0% cholesterol showed significantly higher (P<0.05) specific growth rate (SGR) than the other groups, who suffered from relatively lower molt death syndrome (MDS). Cholesterol content in the serum, whole body, and hepatopancreas increased in relation to dietary cholesterol. Muscle lipid content was significantly higher (P<0.05) in crabs fed the diet with 0.2% cholesterol compared to the other treatments. Crabs fed moderate dietary cholesterol levels showed higher alkaline phosphatase (AKP) or acid phosphatase (ACP) levels than those fed 0.2% or 1.4% cholesterol diets. The present study also showed that dietary cholesterol supplementation generally increased serum superoxide dismutase (SOD) activity. Overall, moderate dietary cholesterol (1.0 %) enhanced the performance of growth, survival, molting, and immunity of juvenile swimming crab P. trituberculatus

Investigation of multi-stage forward osmosis membrane process for concentrating high-osmotic acrylamide solution

Acrylamide is an important chemical in great global demand for the synthesis of polyacrylamide. A facile and benign approach of concentrating high osmotic acrylamide aqueous solution at a low temperature is needed to replace the current energy-intensive and cost-expensive thermal flash evaporation process. For the first time, a multi-stage forward osmosis (FO) process has been developed to concentrate acrylamide solution from 200 g/L to 600 g/L. Thin-film composite (TFC) membrane was fabricated and used for the multi-stage FO process. Acrylamide feed solution (FS) with various concentrations was systematically characterized in terms of viscosity and osmotic pressure (OP). Draw solutes including NaCl and MgCl2 were tested, and their reverse salt fluxes were measured with the quantification of their accumulative contents in the resultant concentrated acrylamide solution. Different operation modes including AL-FS (active layer facing FS) and AL-DS (active layer facing DS) were explored to optimize the system efficiency of the FO concentration process. Both single- and multi-stage FO operations were investigated, and their performances were quantified to assess the efficiency of the concentration of acrylamide solution. The results demonstrate that the multi-stage FO operation could dramatically improve the system efficiency for the concentration of acrylamide solution better than the single-stage FO process, and the draw concentration renewal at Stages II, III, and IV led to a water flux increase of 18.56%, 17.52%, and 18.43%, respectively. Moreover, the accumulated MgCl2 in the final 600 g/L acrylamide solution was below 3.7 g/L, less than 0.62 wt% impurity in the product of acrylamide. Our work provides a practical insight into the viability and optimization of a multi-stage FO process for concentrating high osmotic chemicals

A Highly Efficient Synthetic Vector: Nonhydrodynamic Delivery of DNA to Hepatocyte Nuclei in Vivo

Multifunctional membrane-core nanoparticles, composed of calcium phosphate cores, arginine-rich peptides, cationic and PEGylated lipid membranes, and galactose targeting ligands, have been developed as synthetic vectors for efficient nuclear delivery of plasmid DNA and subsequent gene expression in hepatocytes in vivo. Targeted particles exhibited rapid and extensive hepatic accumulation and were predominantly internalized by hepatocytes, while the inclusion of such peptides in LCP was sufficient to elicit high degrees of nuclear translocation of plasmid DNA. Monocyclic CR8C significantly enhanced in vivo gene expression over ten-fold more than linear CR8C, likely due to a release-favoring mechanism of the DNA/peptide complex. Though 100-fold lower in activity than that achieved via hydrodynamic injection, this formulation presents as a much less invasive alternative. To our knowledge, this is the most effective synthetic vector for liver gene transfer