Characterization of neuronal differentiation in human adipose-derived stromal cells: morphological, molecular, and ultrastructural insights.

OBJECTIVE: Adipose-derived stromal cells (ADSCs) have shown promise as a potential source of neural differentiation. In this study, we investigated the morphological, molecular and ultrastructural features of ADSCs during neuronal differentiation. METHODS: ADSCs were induced in vitro and their differentiation was examined at different time points. Immunocytochemical staining was performed to detect the expression of neuron-specific markers NSE and MAP-2. Immunofluorescence double labeling and Western blot detected the co-expression of presynaptic markers (CaMKII, SynCAM1, SYN) and postsynaptic markers (PSD-95, Synapsin I). Scanning electron microscopy (SEM) was performed to detect the synaptic structural features of differentiated neurons. RESULTS: ADSCs showed diverse morphological features during differentiation, gradually acquiring a neuron-like spindle shape and organized arrangement. The expression of neuron-specific markers and synaptic markers peaked at 5h of induction. Scanning electron microscopy showed polygonal protrusions of ADSC-derived neurons, and transmission electron microscopy showed characteristic ultrastructures such as nidus, synaptic vesicle-like structures, and tight junctions. CONCLUSION: Our findings suggest that ADSCs differentiated for 5h have neuronal features, including morphological, molecular, and ultrastructural resemblance to neurons, as well as the formation of synaptic structures. These insights contribute to a better understanding of ADSC-based neuronal differentiation and pave the way for future applications in regenerative medicine and neurodegenerative diseases.

Preparation of an Aminated Lignin/Fe(III)/Polyvinyl Alcohol Film: A Packaging Material with UV Resistance and Slow-Release Function.

To reduce the usage of petroleum-based plastic products, a lignin-based film material named aminated lignin/Fe(III)/PVA was developed. The mixture of 8 g lignin, 12 mL diethylenetriamine, 200 mL NaOH solution (0.4 mol·L-1), and 8 mL formaldehyde was heated at 85 °C for 4 h; after the aminated lignin was impregnated in the Fe(NO3)3 solution, a mixture of 3 g aminated lignin/Fe(III), 7 g PVA, and 200 mL NaOH solution (pH 8) was heated at 85 °C for 60 min; after 2 mL of glycerin was added, the mixture was spread on a glass plate to obtain the aminated lignin/Fe(III)/PVA film. This film demonstrated hydrophobicity, an UV-blocking function, and a good slow-release performance. Due to the formation of hydrogen bonds between the hydroxyl groups of lignin and PVA, the tensile strength, the elongation at break, and the fracture resistance of the film were 9.1%, 107.8%, and 21.9% higher than that of pure PVA film, respectively. The iron content of aminated lignin/Fe(III)/PVA was 1.06 wt%, which mainly existed in a trivalent form. The aminated lignin/Fe(III)/PVA film has the potential to be used as a food packaging material with anti-ultraviolet light function and can also be developed as other packaging materials, such as seedling bowls, pots for transplanting, and coating films during transport.

Time Scaling for In Vitro-In Vivo Correlation: the Inverse Release Function (IRF) Approach.

In vitro-in vivo correlations (IVIVC) are methods used to create a link between biopharmaceutical properties such as dissolution and physiological response such as plasma concentration. Level A IVIVC defines 1:1 relationship between the percent absorbed in vivo and the percent dissolved in vitro. A successful level A IVIVC provides the capacity to predict in vivo behavior based only on in vitro data with application in formulation development and support of biowaivers recognized by regulatory agencies across the world. Level A regression may be complicated due to differences in time scales as well as the lack of coincident times of similar release in vitro and in vivo leading to approximate time-to-time links and subsequent loss of information. Here, a novel method to establish Levy's plot and to provide time scaling for improved IVIVC predictive capacity is presented. The method is mathematically closed and is an inverse release function (IRF) characterizing the single (or more) phases of dissolution/absorption. It uses the complete set of information available from all time points both in vitro and in vivo. An extended-release formulation development situation is presented with three increasing release rate test products compared in a trial versus a reference product. First, the standard level A regression was made. Prediction errors for internal validation were higher than 10% for Cmax. The IRF method was applied to obtain the in vitro times of percentage dissolved equivalent to percentage absorbed. The prediction errors from the IRF level A correlation were nearly negligible.

Small caliber arterial endothelial cells calcium signals elicited by PAR2 are preserved from endothelial dysfunction.

Endothelial cell (EC)-dependent vasodilation by proteinase-activated receptor 2 (PAR2) is preserved in small caliber arteries in disease states where vasodilation by muscarinic receptors is decreased. In this study, we identified and characterized the PAR2-mediated intracellular calcium (Ca(2+))-release mechanisms in EC from small caliber arteries in healthy and diseased states. Mesenteric arterial EC were isolated from PAR2 wild-type (WT) and null mice, after saline (controls) or angiotensin II (AngII) infusion, for imaging intracellular calcium and characterizing the calcium-release system by immunofluorescence. EC Ca(2+) signals comprised two forms of Ca(2+)-release events that had distinct spatial-temporal properties and occurred near either the plasmalemma (peripheral) or center of EC. In healthy EC, PAR2-dependent increases in the densities and firing rates of both forms of Ca(2+)-release were abolished by inositol 1,4,5- trisphosphate receptor (IP3R) inhibitor, but partially reduced by transient potential vanilloid channels inhibitor ruthenium red (RR). Acetylcholine (ACh)-induced less overall Ca(2+)-release than PAR2 activation, but enhanced selectively the incidence of central events. PAR2-dependent Ca(2+)-activity, inhibitors sensitivities, IP3R, small- and intermediate-conductance Ca(2+)-activated potassium channels expressions were unchanged in EC from AngII WT. However, the same cells exhibited decreases in ACh-induced Ca(2+)-release, RR sensitivity, and endothelial nitric oxide synthase expression, indicating AngII-induced dysfunction was differentiated by receptor, Ca(2+)-release, and downstream targets of EC activation. We conclude that PAR2 and muscarinic receptors selectively elicit two elementary Ca(2+) signals in single EC. PAR2-selective IP3R-dependent peripheral Ca(2+)-release mechanisms are identical between healthy and diseased states. Further study of PAR2-selective Ca(2+)-release for eliciting pathological and/or normal EC functions is warranted.

Study on the correlation between the early phase insulin secretion index and 72 hours continuous glucose levels in patients of impaired glucose tolerance / 中国医师进修杂志

Objective To investigate the correlation between the early phase insulin secretion index and 72 h continuous glucose levels in patients of impaired glucose tolerance (IGT). Methods According to repeated 75 g oral glucose tolerance test (75 g OGTT) ,62 cases were divided into 2 groups: normal glucose tolerance group (NGT group, 30 cases) and isolated impaired glucose tolerance group (IGT group, 32 cases). Insulin levels were detected and HOMA-IR,HOMA-β , ΔI30/ΔG30,AUCI were calculated. The blood glucose levels were monitored by continuous glucose monitoring system for 72 h. The characteristics of postprandial glucose excursion were studied based on peak postprandial glucose (PPC) concentration, time to PPG (Δt) , postprandial glucose excursion (PPGE) and duration of postprandial glucose excursion (DPE). They were statistically analyzed by SPSS12.0. Results The levels of PPG and PPGE were significantly higher in IGT group (P < 0.05). Δt and DPE delayed obviously in IGT group (P < 0.05). HOM A-IR in IGT group was higher than that in NGT group (1.68 ± 1.03 vs 1.15 ± 0.90, P < 0.01), Δ I30/ΔG30 and HOMA- β was significantly lower in IGT group than that in NGT group (3.85 ± 1.04 vs 6.42 ±1.05,52.97 ± 2.02 vs 55.68 ± 12.45, P < 0.01 or < 0.05). Conclusions Higher postprandial glucose levels are characteristics of IGT patients,and the function of islet β cell after glucose load is impaired more severely. The levels of FPG and 2hPG are positively correlated with insulin resistance, and negatively correlated with islet β cell function.