Solubility-permeability interplay of a supersaturated lutein delivery system constructed by glycosylated stevioside and hydroxypropyl-methylcellulose.

This study investigated the solubilizing capacity of glycosylated stevioside/hydroxypropyl-methylcellulose (stevia-G-HPMC) complexes with varying mass ratios on lutein. The impact on the steady-state flux and permeability coefficient of intracellular lutein was also explored through the construction of a Caco-2 cellular transport model. The results indicated that the equilibrium solubility of lutein linearly increased with an increase in stevia-G amount. The stability constants of the ternary system surpassed those of the binary system. Molecular dynamics simulation revealed a tight and stable structure in lutein supersaturated complexes. Meanwhile, lutein-stevia-G-HPMC complexes demonstrated superior cumulative penetrations, with the peak Papp (AP â†’ BL) value being (3.24 ± 0.89) × 10-5 cm·s-1. There was a slight decrease in Papp (BL â†’ AP), which improved the forward transport of lutein. Highly soluble lutein in aqueous environments saturated the extracellular transport proteins on the AP side of cell membranes, thereby maintaining the high permeability transport. Notably, the permeability trend of lutein in Caco-2 cells negatively correlated with the equilibrium solubility and matched the single exponential growth model. When the mass ratio of lutein, stevia-G and HPMC was 1:21:5, the solubility-permeability trade-off of lutein was effectively maintained.

Phytochemicals in Chronic Disease Prevention.

Chronic diseases, also known as noncommunicable diseases (NCD), are characterized by long durations and a slow progression of the associated medical conditions [...].

LIP5, a MVB biogenesis regulator, is required for rice growth.

LYST-INTERACTING PROTEIN5 (LIP5) is a conserved regulator of multivesicular body (MVB) biogenesis in eukaryotes. In Arabidopsis, AtLIP5 is a target of stress-responsive MITOGEN-ACTIVATED PROTEIN KINASE3 and 6 and mediates stress-induced MVB biogenesis to promote stress responses. However, Arabidopsis atlip5 knockout mutants are normal in growth and development. Here we report that rice OsLIP5 gene could fully restore both the disease resistance and salt tolerance of the Arabidopsis oslip5 mutant plants to the wild-type levels. Unlike Arabidopsis atlip5 mutants, rice oslip5 mutants were severely stunted, developed necrotic lesions and all died before flowering. Unlike in Arabidopsis, LIP5 regulated endocytosis under both stress and normal conditions in rice. These findings indicate that there is strong evolutionary divergence among different plants in the role of the conserved LIP5-regulated MVB pathway in normal plant growth.

FOXP4 promotes proliferation of human spermatogonial stem cells.

Continuous self-renewal and differentiation of spermatogonial stem cells (SSCs) is vital for maintenance of adult spermatogenesis. Although several spermatogonial stem cell regulators have been extensively investigated in rodents, regulatory mechanisms of human SSC self-renewal and differentiation have not been fully established. We analyzed single-cell sequencing data from the human testis and found that forkhead box P4 (FOXP4) expression gradually increased with development of SSCs. Further analysis of its expression patterns in human testicular tissues revealed that FOXP4 specifically marks a subset of spermatogonia with stem cell potential. Conditional inactivation of FOXP4 in human SSC lines suppressed SSC proliferation and significantly activated apoptosis. FOXP4 expressions were markedly suppressed in tissues with dysregulated spermatogenesis. These findings imply that FOXP4 is involved in human SSC proliferation, which will help elucidate on the mechanisms controlling the fate decisions in human SSCs.

Hybridization of glucosyl stevioside and hydroxypropyl methylcellulose to improve the solubility of lutein.

In this paper, a lutein-glucosyl stevioside (stevia-G)-hydroxypropyl methylcellulose (HPMC) complex was prepared via an antisolvent precipitation combined with dynamic high pressure microfluidization method. The solubility, microstructure, crystallinity and thermodynamic properties of the freeze-dried powder were investigated, as well as the formation mechanism and the storage stability of the produced complex. When the optimal mass ratio of lutein, stevia-G, and HPMC was 1: 40: 0.5, the apparent solubility of lutein reached 2805.47 ± 24.94 µg·mL-1, which was approximately 5600 times higher than that of lutein crystals. The lutein-stevia-G-HPMC complex formed an amorphous dispersed structure and was in a thermodynamically high energy state. The self-assembled micelle structure of stevia-G and HPMC polymer created a supersaturated system mainly by multiple hydrogen bonding, which promoted maximum lutein dissolving, delayed supersaturated crystallization process, and hindered precipitation. The present results suggested the complex formed by stevia-G and HPMC effectively promote lutein's hydrophilicity and stability.

Cargo Recognition and Function of Selective Autophagy Receptors in Plants.

Autophagy is a major quality control system for degradation of unwanted or damaged cytoplasmic components to promote cellular homeostasis. Although non-selective bulk degradation of cytoplasm by autophagy plays a role during cellular response to nutrient deprivation, the broad roles of autophagy are primarily mediated by selective clearance of specifically targeted components. Selective autophagy relies on cargo receptors that recognize targeted components and recruit them to autophagosomes through interaction with lapidated autophagy-related protein 8 (ATG8) family proteins anchored in the membrane of the forming autophagosomes. In mammals and yeast, a large collection of selective autophagy receptors have been identified that mediate the selective autophagic degradation of organelles, aggregation-prone misfolded proteins and other unwanted or nonnative proteins. A substantial number of selective autophagy receptors have also been identified and functionally characterized in plants. Some of the autophagy receptors in plants are evolutionarily conserved with homologs in other types of organisms, while a majority of them are plant-specific or plant species-specific. Plant selective autophagy receptors mediate autophagic degradation of not only misfolded, nonactive and otherwise unwanted cellular components but also regulatory and signaling factors and play critical roles in plant responses to a broad spectrum of biotic and abiotic stresses. In this review, we summarize the research on selective autophagy in plants, with an emphasis on the cargo recognition and the biological functions of plant selective autophagy receptors.

Coordination and Crosstalk between Autophagosome and Multivesicular Body Pathways in Plant Stress Responses.

In eukaryotic cells, autophagosomes and multivesicular bodies (MVBs) are two closely related partners in the lysosomal/vacuolar protein degradation system. Autophagosomes are double membrane-bound organelles that transport cytoplasmic components, including proteins and organelles for autophagic degradation in the lysosomes/vacuoles. MVBs are single-membrane organelles in the endocytic pathway that contain intraluminal vesicles whose content is either degraded in the lysosomes/vacuoles or recycled to the cell surface. In plants, both autophagosome and MVB pathways play important roles in plant responses to biotic and abiotic stresses. More recent studies have revealed that autophagosomes and MVBs also act together in plant stress responses in a variety of processes, including deployment of defense-related molecules, regulation of cell death, trafficking and degradation of membrane and soluble constituents, and modulation of plant hormone metabolism and signaling. In this review, we discuss these recent findings on the coordination and crosstalk between autophagosome and MVB pathways that contribute to the complex network of plant stress responses.

Abnormality of Klotho Signaling Is Involved in Polycystic Ovary Syndrome.

This study investigated the involvement of the klotho-associated signaling in the apoptosis of granulosa cells (GCs) from the ovaries of patients with polycystic ovary syndrome (PCOS) and PCOS animals. Primary GCs were obtained from 26 healthy women and 43 women with PCOS. The PCOS animal model was established by the injection of dehydroepiandrosterone (DHEA). Klotho protein and associated microRNA expression in human primary GCs and rats' ovarian tissues were measured by Western blot and real-time polymerase chain reaction, respectively. Results showed that significantly lower miR-126-5p and miR-29a-5p microRNA expressions, higher klotho protein expression, lower insulin growth factor 1 (IGF-1R) and Wnt family member 1 (Wnt1) protein expressions, and lower Akt phosphorylation at Ser473 and Thr308 residues were observed in the GCs from patients with PCOS and the ovarian tissues of PCOS rats compared to that in GCs from healthy women and ovarian tissues of normal control rats, respectively. Knockdown of klotho gene expression normalized IGF-1R and Wnt1 protein expressions and Akt phosphorylation in GCs from patients with PCOS and the ovarian tissues from PCOS rats; it also blocked the effects of insulin on apoptosis and proliferation in GCs from patients with PCOS and inhibited caspase-3 activity in ovarian tissues of PCOS rats. Knockdown of klotho gene expression increased the pregnancy rate in DHEA-treated female rats and increased the body weight of their newborns through normalizing the ovarian function and decreasing the formation of cystic follicles. In conclusion, the miR-126-5p, miR-29a-5p/klotho/insulin-IGF-1, Wnt, and Akt signal pathway may be involved in the apoptosis of GCs and subsequent development of PCOS.

P1 and N170 components distinguish human-like and animal-like makeup stimuli.

This study used event-related potentials to investigate the sensitivity of P1 and N170 components to human-like and animal-like makeup stimuli, which were derived from pictures of Peking opera characters. As predicted, human-like makeup stimuli elicited larger P1 and N170 amplitudes than did animal-like makeup stimuli. Interestingly, a right hemisphere advantage was observed for human-like but not for animal-like makeup stimuli. Dipole source analyses of 130-200-ms window showed that the bilateral fusiform face area may contribute to the differential sensitivity of the N170 component in response to human-like and animal-like makeup stimuli. The present study suggests that the amplitudes of both the P1 and the N170 are sensitive for the mouth component of face-like stimuli.

Maintenance of undifferentiated state of human embryonic stem cells in chemical defined medium at high clone density without exogenous cell factors.

OBJECTIVE: To establish human embryonic stem cells (hESCs) feeder-independent and cell factor-free culture system. METHODS: Effect of high and low clone densities of hESCs culture was compared and impact of the clone densities to hESCs culture media was analyzed. RESULTS: HESCs could maintain their undifferentiated states at high clone density (34 clones/cm²) without cell factors. At the same time, the bone morphology protein (BMP)-like induction of N2 and B27 supplements (NB) medium could be modulated by the clone density, and high level of BMP-like induction was accompanied by high clone density. CONCLUSION: High clone density of hESCs can change the environments by themselves to maintain the undifferentiated states, which provides a new clue to explore the mechanism of undifferentiated states of hESCs and simplify the culture medium.

Tumor progression of culture-adapted human embryonic stem cells during long-term culture.

Human embryonic stem cells (hESCs) during long-term culture acquire chromosomal changes similar to those occurring in tumorigenesis. This was raised concerns about the progression from hESCs to malignant cells. This study aimed to investigate the changes in chromosomes, cell phenotype, and genes in culture-adapted hESCs to ascertain whether tumorigenic transformation occurred. By cytogenetic analysis we found progressive karyotypic changes from simple to complex in chHES-3, one of the hESC lines established in our laboratory, during a long-term suboptimal culture. We further compared chHES-3 cells at different karyotypic stages in cell surface markers, in vivo differentiation, cell cycle, apoptosis, and gene expression profiles. We found that the karyotypically aberrant chHES-3 had higher S-phase fraction in cell cycle distributions and antiapoptosis ability. In vivo differentiation of karyotypically normal chHES-3 resulted in relatively mature teratoma, whereas karyotypically aberrant chHES-3 formed immature teratoma (grade III), in which more primary neural epithelium was revealed by pathological analysis. The microarray analysis and real-time PCR results showed that some oncogenes were upregulated in karyotypically aberrant chHES-3 cells, whereas the genes related to differentiation were downregulated, and that Wnt signal pathway was activated. In conclusion, chHES-3 cells underwent deregulation of self-renewal and dysfunction of related genes in long-term culture adaptation, leading to malignant transformation.

Cloning, characterization and primary function study of a novel gene, Cymg1, related to family 2 cystatins.

Cystatins are cysteine proteinase inhibitors. We found two expression sequence tags (ESTs), CA463109 and AV042522, from a mouse testis library using Digital differential display (DDD). By electrical hybridization, a novel gene, Cymg1 (GenBank accession No. AY600990), which has a full length of 0.78 kb, and contains four exons and three introns, was cloned from a mouse testis cDNA library. The gene is located in the 2G3 area of chromosome 2. The full cDNA encompasses the entire open reading frame, encoding 141 amino acid residues. The protein has a cysteine protease inhibitor domain that is related to the family 2 cystatins but lacks critical consensus sites important for cysteine protease inhibition. These characteristics are seen in the CRES subfamily, which are related to the family 2 cystatins and are expressed specifically in the male reproductive tract. CYMG1 has a 44% (48/108) identity with mouse CRES and 30% (42/140) identity with mouse cystatin C. Northern blot analysis showed that the Cymg1 is specifically expressed in adult mouse testes. Cell location studies showed that the GFP-tagged CYMG1 protein was localized in the cytoplasm of HeLa cells. Immunohistochemistry revealed that the CYMG1 protein was expressed in mouse testes spermatogonium, spermatocytes, round spermatids, elongating spermatids and spermatozoa. RT-PCR results also showed that Cymg1 was expressed in mouse testes and spermatogonium. The Cymg1 expression level varied in different developmental stages: it was low 1 week postpartum, steadily increased 2 to 5 weeks postpartum, and was highest 7 weeks postpartum. The expression level at 5 weeks postpartum was maintained during 13 to 57 weeks postpartum. The Cymg1 expression level in the testes over different developmental stages correlates with the mouse spermatogenesis and sexual maturation process. All these indicate that Cymg1 might play an important role in mouse spermatogenesis and sexual maturation.

Newly expressed proteins of mouse embryonic fibroblasts irradiated to be inactive.

It has been found that post-radiation mouse embryonic fibroblasts can well maintain the pluripotency in human embryonic stem cells. However, the molecular mechanism remains unclear. In the present study, the new protein expression profile of post-radiation mouse embryonic fibroblasts was analyzed by immobilized pH gradient 2-dimensional polyacrylamide gel electrophoresis. Image analysis following silver staining revealed (969+/-57) vs. (1085+/-107) spots from post-radiation mouse embryonic fibroblasts and pre-radiation ones, respectively. Some newly expressed proteins, which were only abundantly present after irradiation, were subjected to peptide mass fingerprint analysis and identified using MALDI-TOF-MS, SWISS-PROT database, and RT-PCR. Several of those proteins were preliminarily identified to participate in cytokine secretion, cell signal transduction, transcriptional regulation, and apoptosis, etc., which suggested that inactive post-radiation mouse embryonic fibroblasts expressed some new proteins that may underlie the molecular mechanisms to maintain the pluripotency in human embryonic stem cells.

[Isolation culture and identification of human neural stem cells from human embryos].

OBJECTIVE: To obtain and culture the human neural stem cells from the aborted human embryos. METHODS: The neural stem cells were isolated and purified by the specific proliferous culture system of neural stem cells, and the molecular maker and multi-potency of the obtained neural stem cells were identified. RESULTS: Human neural stem cells, which were isolated from 8 - 10 month old aborted human embryos, could express nestin ( a kind of specified antigen of the neural stem cell), and it could be differentiate into neurons and glials. CONCLUSION: Neural stem cells can exist in human embryos, and it can be expanded in vitro.