Peptidome comparison on the immune regulation effects of different casein fractions in a cyclophosphamide mouse model.

The protein composition of human milk plays a crucial role in infant formula milk powder formulation. Notably, significant differences exist between bovine casein and human milk casein. Previous studies have shown that casein hydrolysates could enhance immune function; however, gastrointestinal dyspepsia in infants affects the type and function of peptides. Therefore, the present study used peptidomics to sequence and analyze hydrolyzed peptides from different casein fractions. Additionally, animal experiments were conducted to assess the functionality of these casein fractions and elucidate their differences. The results revealed variations in peptide composition among the different casein fractions of formula milk powder. Interestingly, milk powder formulated with both ß- and κ-casein (BK) exhibited significant enrichment of peptides related to the immune system. Moreover, the BK group significantly alleviated immune organ damage in cyclophosphamide-treated mice and regulated serum levels of pro-inflammatory and anti-inflammatory factors. Furthermore, feeding different casein fractions influenced the intestinal microflora of cyclophosphamide-treated mice, with the BK group mitigating the changes caused by cyclophosphamide. In conclusion, the findings suggest that BK formula in milk powder has the potential to positively enhance immunity. This study provides a robust theoretical basis for human-emulsified formula milk powder development.

In vitro skin retention and drug permeation study of Tongluo-Qutong rubber plaster by UPLC/UV/MS/MS

Abstract Tongluo-Qutong rubber plaster (TQRP), a typical Chinese patent medicine that contains 13 different herbal remedies, is widely used in clinical practice for the treatment of cervical spondylosis and osteoarthritis. However, due to a lack of in vitro transdermal studies, the active ingredients of TQRP have not been fully elucidated. This presents a huge obstacle for quality evaluation, pharmacokinetic studies and clinical safety assessment of TQRP. In this work, a UPLC/UV/MS/MS method was established and validated to evaluate five analytes in TQRP. The validation demonstrated linearity (r > 0.99), specificity (no co-eluting peaks at the retention times of the analytes), and precision (RSD < 15%) within acceptable parameters. A skin permeation study was performed to determine the concentrations of drugs delivered to the dermis. The 24-hour cumulative permeation of ferulic acid, aleo-emodin, emodin and piperine were 303.68, 709.31, 671.06 and 25561.01 ng/cm2, respectively. According to the fitting data of the TQRP active components, skin permeation was mainly due to a combination of passive diffusion and drug release after matrix erosion

Revalorization of sunflower stalk pith as feedstock for the coproduction of pectin and glucose using a two-step dilute acid pretreatment process.

BACKGROUND: Sunflower stalk pith, residue from the processing of sunflower, is rich in pectin and cellulose, thereby acting as an economic raw material for the acquisition of these compounds. In order to increase the commercial value of sunflower processing industry, a two-step dilute sulfuric acid treatment process was conducted on spent sunflower stalk pith to obtain the value-added products, pectin and glucose. RESULTS: In this study, pectin was firstly extracted under mild acid condition to avoid pectin degradation, which was conducted at 90 °C with a pH of 2.0 for 2 h, and ~0.14 g/g of pectin could be recovered. Then the remaining solids after pectin extraction were subjected to the reinforced treatment process with 0.75% H2SO4 at 150 °C for 30 min to further improve enzymatic hydrolysis efficiency. Moreover, by combining a fed-batch enzymatic hydrolysis strategy, a solid loading content of 16% was successfully achieved and the glucose titer reached 103.1 g/L with a yield of 83.6%. CONCLUSION: Finally, ~140 g pectin and 260 g glucose were produced from 1 kg of raw sunflower stalk pith using the integrated biorefinery process. This work puts forward a two-step dilute acid pretreatment combined with enzymatic hydrolysis method to produce pectin and glucose from sunflower spent waste.

A Remixed-Fermentation Technique for the Simultaneous Bioconversion of Corncob C6 and C5 Sugars to Probiotic Bacillus subtilis.

The probiotic strain of Bacillus subtilis presents a promising application potential for the value-added bio-utilization of lignocellulosic carbohydrates. By the combined acidolysis pretreatment and enzymatic hydrolysis, hemicellulose and cellulose constituents of corncob were efficiently converted respectively into fermentable C5 and C6 sugars, mainly including xylose and glucose. B. subtilis grew well in xylose solution while it was hindered completely in the acidolysis broth because of the bio-toxicity of degraded chemicals derived from corncob. A mixed-fermentation technique was therefore developed and performed to blend the acidolysis broth and enzymatic hydrolysis slurry together, by which C5 and C6 sugar molecules were successfully fermented and efficiently utilized for the growth of B. subtilis cells with a yield of 0.33 g cells/g sugar consumed. A net amount of 205.9 ± 9.0 g of B. subtilis powder was obtained from 1000 g of corncob that could improve the economic benefits of the process to around 5-7 times.

A strategy to improve the oral availability of baicalein: The baicalein-theophylline cocrystal.

Baicalein (BE) is a flavonoid compound derived from the roots of Scutellaria baicalensis. It has widely been used as anti-oxidant, anti-virus, anti-bacteria, anti-inflammatory and anti-allergic therapies. Due to its poor water solubility (16.82 µg/ml), the therapeutic effectiveness and oral bioavailability of Baicalein are highly limited. The purpose of this study was to investigate the possibility of baicalein-theophylline (BE-TH) cocrystals to achieve the simultaneous enhancement in dissolution and oral bioavailability of BE. The cocrystal had the new characteristic of scanning electron microscopy, differential scanning calorimetry thermograms and X-ray powder diffraction. Compared with coarse powder of BE, BE-TH cocrystals had significantly improved the solubility of BE. The dissolution test showed that the BE-TH cocrystals demonstrated 2.2-fold and 7.1-fold higher rate of dissolution than that of BE coarse powder in HCl (pH = 1.2) and phosphate buffer (PBS, pH = 6.8), respectively. Moreover, the cocrystals exhibited a 5.86-fold higher area under the curve in rats after the oral administration. This investigation enriched the present understanding of cocrystals on their behavior in vitro and in vivo, and built the groundwork for future development of BE as a promising compound into efficacious drug products.

Establishment, characterization and immortalization of a fibroblast cell line from the Chinese red belly toad Bombina maxima skin.

The skin of the amphibian Bombina maxima is rich in biologically active proteins and peptides, most of which have mammalian analogues. The physiological functions of most of the mammalian analogues are still unknown. Thus, Bombina maxima skin may be a promising model to reveal the physiological role of these proteins and peptides because of their large capacity for secretion. To investigate the physiological role of these proteins and peptides in vitro, a fibroblast cell line was successfully established from Bombina maxima tadpole skin. The cell line grew to form a monolayer with cells of a uniform shape and abundant rough endoplasmic reticulum, which are typical characteristics of fibroblasts. Further identification at a molecular level revealed that they strongly expressed the fibroblast marker protein vimentin. The chromosome number of these cells is 2n = 28, and most of them were diploid. Growth property analysis showed that they grew well for 14 passages. However, cells showed decreased proliferative ability after passage 15. Thus, we tried to immortalize the cells through the overexpression of SV40 T antigen. After selecting by G418, cells stably expressed SV40 large T antigen and showed enhanced proliferative ability and increased telomerase activity. Signal transduction analysis revealed functional p42 mitogen-activated protein (MAP) kinase in immortalized Bombina maxima dermal fibroblasts. Primary fibroblast cells and the immortalized fibroblast cells from Bombina maxima cultured in the present study can be used to investigate the physiological role of Bombina maxima skin-secreted proteins and peptides. In addition, the methods for primary cell culturing and cell immortalization will be useful for culturing and immortalizing cells from other types of amphibians.

[Detection and comparison of physiological indexes in the wild and laboratory tree shrew].

To provide fundamental basis for the tree shrew models of human diseases, we examined and compared the physiological and biochemical indexes between wild and laboratory tree shrews. Blood samples were taken from 54 wild tree shrews that were housed in laboratory for 1-2 months, and from 54 first-generation of the laboratory tree shrews; each group had nearly equal male and female composition. Some of the first reported physiological and biochemical indexes were showed no significant differences between genders, and these indexes in laboratory tree shrews were as follows [medium (inter-quartile range) ]: CK 1449 (956) U/L, CTNI 5.94 (7.23) ug/L, TBA15.6 (19.7) µmol/L, FRUC 393.5 (80.8) µmol/L and LDL-C0.36 (0.32); and in the wild tree shrews, 986 (564) U/L, 4.01 (4.10) µg/L, 20.0 (20.6) µmol/L, 379.0 (104.0) µmol/L and 0.46 (0.23) mmol/L, respectively. In the laboratory tree shrews, the variations of physiological and biochemical indexes were smaller, but the mean values of some indicators related to liver and heart functions became higher. These data would be valuable for the development of tree shrew models of human diseases.