CLDN5: From structure and regulation to roles in tumors and other diseases beyond CNS disorders.

Claudin-5 (CLDN5) is an essential component of tight junctions (TJs) and is critical for the integrity of the blood-brain barrier (BBB), ensuring homeostasis and protection from damage to the central nervous system (CNS). Currently, many researchers have summarized the role and mechanisms of CLDN5 in CNS diseases. However, it is noteworthy that CLDN5 also plays a significant role in tumor growth and metastasis. In addition, abnormal CLDN5 expression is involved in the development of respiratory diseases, intestinal diseases, cardiac diseases, and diabetic ocular complications. This paper aims to review the structure, expression, and regulation of CLDN5, focusing on its role in tumors, including its expression and regulation, effects on malignant phenotypes, and clinical significance. Furthermore, this paper will provide an overview of the role and mechanisms of CLDN5 in respiratory diseases, intestinal diseases, cardiac diseases, and diabetic ocular complications.

Comprehensive evaluation of aroma and taste properties of different parts from the wampee fruit.

Wampee is a tropical fruit having high medicinal value. To fully realize the fruit's potential, it is essential to reveal the flavor characteristics. In this study, a comprehensive analysis of the aroma and taste profiles of different parts from the wampee fruit was conducted. The aroma profile was analyzed by E-nose, and 67 volatile components were identified through HS-SPME-GC-MS. Among them, 11 were considered as crucial compounds. Additionally, 42 volatile components were identified by HS-GC-IMS, with 22 compounds showing a variable importance in projection scores greater than 1.0. Moreover, the taste profile and representative compounds were analyzed by E-tongue and HPLC, and 12 compounds were considered as important taste contributors based on taste activity value. These findings shed light on the various compounds responsible for the unique aroma and taste of the wampee fruit, providing theoretical foundation for exploring ways for its comprehensive utilization and development.

Nucleotides and free amino acids in sea urchin Paracentrotus lividus gonads: Contributions for freshness and overall taste.

This study aims to report the chemical changes, specifically the nucleotides and free amino acids (FAAs) during refrigerated storage of live sea urchin Paracentrotus lividus (7 °C) and packed gonads (2 °C) and respective contributions to overall taste. Results showed that the adenylate energy charge (AEC) is an adequate indicator for live sea urchin through storage, while K-value does not distinguish freshness. Changes of both indexes were not clear in packed gonads. The FAAs profile were related with maturation stage where amino acids associated to sweet (Gly, Ala), umami (Glu) and bitter (Arg, Lys, His, Val) had significant contribution to overall taste. In general, the storage did not significantly affect the FAAs content nor its contribution to overall taste in contrast to nucleotides that seemed to induce potential changes after day 5. The findings provide insights that will complement a full study on quality index scheme development of the products.

Development, validation, and uncertainty measurement of HPLC-DAD method for determination of some free amino acids in infant formula and medical food products for inborn errors of metabolism.

This research aims at determining some free amino acids in amino acid-based infant formulas and amino acid-modified medical foods for inborn errors of metabolism to prove their quality. A method based on high-performance liquid chromatography and diode array detection was developed and validated. Then, overall uncertainty was estimated by the bottom-up approach. Applying the weighted least squares regression method suggested good linearity with coefficient of determinations ≥ 0.9960. The limits of detection were calculated between 0.01 and 0.28 µg/mL. The most repetitive recovery values were obtained in the range of 91-108%, with RSDs ≤ 15%. The expanded uncertainties were below 20% for most amino acids. The contributions of linear regression and repeatability are two main factors in estimating overall uncertainty. The results offer this method as a simple and easy procedure for analyzing free amino acids in seven powdered medical foods designed for phenylketonuria, maple syrup urine disease, methylmalonic, and propionic acidemia.

Microcrystalline cellulose as an effective crystal growth inhibitor for the ternary Ibrutinib formulation.

The objective of present study is to explore whether polysaccharide could be a crystal growth inhibitor in poorly soluble antitumor drug Ibrutinib (IBR) formulation. In this work, small molecular ligands (amino or organic acids) in co-amorphous system (CAS) were preliminarily screened. A polysaccharide, microcrystalline cellulose (MCC) was selected to stabilize amorphous drug and enhance pharmacokinetic properties. Fourier-transform infrared, Raman, and X-ray photoelectron spectroscopy confirmed the ionic interaction of the ternary IBR formulation. Moreover, the biosafety of the ternary formulation was the same as that of IBR while the in vitro performance advantage of the ternary formulation was converted into higher bioavailability in vivo experiments. Overall, MCC as an effective crystal growth inhibitor in the novel ternary IBR formulation is a promising approach to improve the dissolution rate of crystalline drugs and the stability of amorphous drugs, as well as providing a theoretical basis for clinical applications.

Investigation of intraregional variation, grape amino acids, and pre-fermentation freezing on varietal thiols and their precursors for Vitis vinifera Sauvignon blanc.

Sauvignon blanc grape samples (n = 21) from across a single Geographical Indication of South Australia were analysed for thiol precursors and amino acids, and fermented in an identical laboratory-scale fermentation trial to investigate the intraregional pattern of varietal thiols in the wines. Precursors and thiols exhibited obvious intraregional diversity, and notably, stronger correlations were observed between a number of amino acids and thiol precursors (especially with glutamic acid, r ≤ -0.73) rather than free thiols. Additionally, pre-fermentation freezing (-20 °C, 1 month) was applied to five selected fresh grape samples and their juices, followed by identical fermentation. In comparison to wines from fresh grapes or frozen juices, significant elevation of varietal thiols (up to 10-fold) occurred in the wines derived from frozen grapes, with parallel increases of precursors (up to 19-fold) in juices from frozen berries. These novel results may lead to new strategies for thiol enhancement during winemaking.

Integration of proteomics and metabolomics data of early and middle season Hass avocados under heat treatment.

Ripening heterogeneity of Hass avocados results in inconsistent quality fruit delivered to the triggered and ready to eat markets. This research aimed to understand the effect of a heat shock (HS) prior to controlled atmosphere (CA) storage on the reduction of ripening heterogeneity. HS prior to CA storage reduces more drastically the ripening heterogeneity in middle season fruit. Via correlation network analysis we show the different metabolomics networks between HS and CA. High throughput proteomics revealed 135 differentially expressed proteins unique to middle season fruit triggered by HS. Further integration of metabolomics and proteomics data revealed that HS reduced the glycolytic throughput and induced protein degradation to deliver energy for the alternative ripening pathways. l-isoleucine, l-valine, l-aspartic and ubiquitin carboxyl-terminal hydrolase involved in protein degradation were positively correlated to HS samples. Our study provides new insights into the effectiveness of HS in synchronizing ripening of Hass avocados.

Enhancement of γ-aminobutyric acid, avenanthramides, and other health-promoting metabolites in germinating oats (Avena sativa L.) treated with and without power ultrasound.

Power ultrasound as an emerging processing technology has been investigated for stimulating seeds to enhance germination and accumulation of health-promoting metabolites, such as γ-aminobutyric acid (GABA) and phenolic compounds. This work was undertaken to evaluate the effects of power ultrasound (25 kHz) on the nutritional properties of germinated oats, and the microstructure of oat groats after treatment. The changes in the external and internal microstructures of the ultrasound-treated oats kernel were investigated using Environmental Scanning Electron Microscopy (ESEM) and 3D X-ray Micro Computed Tomography (Micro-CT). Physicochemical properties of oats including GABA, free sugars, avenanthramides, total phenolic content, and antioxidant capacities were enhanced after germination. Furthermore, the power ultrasound treatment for 5 min after soaking significantly enhanced the GABA (48-96 h), alanine (24-96 h), succinic acid (48-72 h), total phenolic content (24 h), and total avenanthramides (24 h) in the germinated oats.

Umami taste and its association with energy status in harvested Pleurotus geesteranus stored at different temperatures.

Pleurotus geesteranus has recently been gaining popularity due to its strong umami taste. In the present study, umami taste, energy level, and energy metabolism-related enzymes activity in harvested P. geesteranus, stored at 20, 10, 5, and 0 °C, were investigated to evaluate the relationship between umami taste and energy status. Results showed that the mushroom at 5 °C exhibited significantly higher (p < 0.05) equivalent umami concentration (EUC), higher content of adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP), and higher activity of succinic dehydrogenase (SDH) and cytochrome c oxidase (CCO) in late storage. AMP, associating umami taste with energy, presented a significantly positive correlation with EUC and umami determined by electronic tongue at 5 °C. Furthermore, there were better correlations between umami taste and energy status of mushroom at 5 °C. The results suggest that higher energy status of post-harvest P. geesteranus contributes to better umami taste.

The effect of cooking on umami compounds in wild and cultivated mushrooms.

The effect of cooking on the taste compounds of five mushroom species Agaricus bisporus, Lactarius trivialis, Cantharellus cibarius, Cantharellus tubaeformis and Suillus variegatus were assessed with a special focus on the compounds responsible for the umami taste. Liquid chromatography was used to analyse free amino acids (FAAs) and 5'-nucleotides from fresh and sous vide (SV) cooked mushroom samples and cooking juice. The SV technique enabled analysis of entire mushrooms, including the liquid released during cooking. FAA content decreased when cooking temperature increased, indicating their further chemical reactions. S. variegatus contained highest concentrations of FAAs in analysed wild mushrooms. The umami-enhancing nucleotide 5'-GMP was detected only in cooked samples, concentration being likely dependent on enzymatic activity. The highest concentration of 5'-GMP was detected in cooked L. trivialis samples (17 mg/100 g fw). To our knowledge, the analysed taste compounds of L. trivialis and S. variegatus are documented for the first time.

Metabolic variation in the pulps of two durian cultivars: Unraveling the metabolites that contribute to the flavor.

Durian (Durio zibethinus M.) is a major economic fruit crop in Thailand. In this study, two popular cultivars, namely Chanee and Mon Thong, were collected from three orchards located in eastern Thailand. The pulp metabolome, including 157 annotated metabolites, was explored using capillary electrophoresis-time of flight/mass spectrometry (CE-TOF/MS). Cultivars and harvest years had more impact on metabolite profile separation than cultivation areas. We identified cultivar-dependent metabolite markers related to durian fruit quality traits, such as nutritional value (pyridoxamine), odor (cysteine, leucine), and ripening process (aminocyclopropane carboxylic acid). Interestingly, durian fruit were found to contain high amounts of γ-glutamylcysteine (810.3 ±â€¯257.5 mg/100 g dry weight) and glutathione (158.1 ±â€¯80.4 mg/100 g dry weight), which act as antioxidants and taste enhancers. This metabolite information could be related to consumer preferences and exploited for durian fruit quality improvement.

Highly selective colorimetric detection of putrescine in fish products using o-phthalaldehyde derivatization reaction.

A highly selective colorimetric detection method for putrescine on the basis of an optimized derivatization reaction was established. With the aids of o-phthalaldehyde (OPA) and thioglycolic acid (TGA), putrescine was derived to become red derivatives (PUT-RD), the form that can be detected qualitatively and quantitatively by visual inspection and UV-vis spectrophotometer at λmax of 490 nm, respectively. Key parameters affecting the experiment were investigated by one-factor-at-a-time and response surface analysis. At the optimal condition, this colorimetric method achieved good linearity at putrescine concentrations ranging from 0.8 to 200 µM with detection limit of 0.44 µM. The method also has good selectivity when common amino acids and inorganic ions, as well as ethylenediamine, 1,3-propanediamine, 1,5-pentanediamine, and 1,6-hexanediamine were used as interferences. The established colorimetric method was successfully employed for the detection of putrescine in 10 commercial fish products.

Colorimetric aptasensors for determination of tobramycin in milk and chicken eggs based on DNA and gold nanoparticles.

Colorimetric aptasensors were designed for detection of tobramycin (TOB) based on unmodified gold nanoparticles (AuNPs) and single-strand DNA (ssDNA). In the absence of TOB, the DNA aptamer was coated on the surface of AuNPs to keep it against salt-induced aggregation. In the presence of TOB, aptamer will bind with TOB and detach from the surface of AuNPs because of higher affinities between aptamer and TOB. Then less protection of DNA may result in the aggregation of AuNPs by salt and an apparent color change from red to purple-blue. The developed aptasensors showed a high selectivity and sensitivity for TOB detection. The linearity range and the detection limit were 40-200 nM and 23.3 nM respectively. The validity of the procedure and applicability of aptasensors were successfully used to detect TOB in milk and chicken eggs, and the results were excellent in accord with the values obtained by spectrofluorimetric detection.

Effect of high-pressure treatment on taste and metabolite profiles of ducks with two different vinasse-curing processes.

The effect of high-pressure (HP) (0.1, 150 and 300MPa, 15min) on taste profiles of vinasse-cured ducks was investigated; the metabolite profiles were determined using 1H NMR. HP at 150MPa increased the taste intensity of products compared with the controls, while HP at 300MPa did not further improve their taste compared with 150MPa treated samples. The metabonome of vinasse-cured ducks was dominated by 27 metabolites. HP increased amino acids, glucose, alkaloids and organic acids, but decreased inosine monophosphate and its derivatives, compared with the controls. The increments of metabolites in vinasse-dry-cured duck were higher than those in vinasse-wet-cured duck. The change of metabolites could be related to the enzyme activity, the degradations of proteins, sugars and nucleotides, and the permeation from vinasse-curing agents to duck meat. These findings suggest that 150MPa treatment was effective to improve the taste of vinasse-cured duck.

Development and characterization of an α-l-rhamnosidase mutant with improved thermostability and a higher efficiency for debittering orange juice.

The glycoside hydrolase, α-l-rhamnosidase, could remove the bitter taste of naringin from citrus juices. However, most α-l-rhamnosidases are easily deactivated at high temperatures, limiting the practice in debittering citrus juices. The V529A mutant of the α-l-rhamnosidase r-Rha1 from Aspergillus niger JMU-TS528 was developed with improved thermostability using directed evolution technology and site-directed mutagenesis. The enzyme mutant had a half-live of thermal inactivation T(1/2) of 1.92 h, 25.00 min, and 2 min at 60, 65, and 70 °C, respectively. In addition, it had improved substrate affinity and better resistance to the inhibition of glucose. The improved substrate affinity was related to its lowered binding energy. Most significantly, the naringin content was reduced to below the bitter taste threshold by treatment with 75 U/mL of the mutant during the preheating process of orange juice production. The comprehensive results indicate that thermostability improvement could promote the practical value of α-l-rhamnosidase in citrus juice processing.

Cluster shading modifies amino acids in grape (Vitis vinifera L.) berries in a genotype- and tissue-dependent manner.

Amino acid composition of the grape berry at harvest is important for wine making. The present study investigates the complex interplay between tissue, cultivar and light conditions that determine berry amino acid content. Twenty amino acids were assessed in the berry skin and pulp of two grape cultivars (Gamay Noir and Gamay Fréaux), grown under either light exposure or cluster shading conditions. In all samples, cluster shading significantly reduced most amino acids, except gamma-aminobutyric acid (GABA) and phenylalanine. However, the magnitude of the decrease was stronger in the skin (67.0% decrease) than in the pulp (30.4%) and stronger in cv. Gamay Noir (69.7%) than in Gamay Fréaux (30.7%). Cluster shading also significantly modified amino acid composition by decreasing the proline content while increasing the GABA content. These results are of oenological interest for shaping the amino acid composition of the must and improving wine quality.

Complexation of κ-carrageenan with gelatin in the aqueous phase analysed by 1H NMR kinetics and relaxation.

The 1H NMR spectroscopy is used to study the kinetics of gelation in the aqueous mixtures of κ-carrageenan with gelatin. The time dependence of NMR signals intensities shows that the kinetics of gel formation consists of classical 'fast' (rate constant k≈6h-1) and 'slow' (k≈1h-1) periods, corresponding to a coil→helix transition and subsequent aggregation of helices. Upon increase of the κ-carrageenan/gelatin (w/w) ratio Z the rate of the fast process slows down by a factor of 1.6-2.4. Further analysis was done by studying the dependence of spin-spin relaxation times of protons of gelatin on Z in the aqueous phase. A qualitative scheme describing hydrogel formation in the complex solution is given. It is hypothesized that at higher concentration of PECs the hydrogel structure network is stabilized by three types of nodes: triple helices of gelatin and intra-/inter-molecular double helices of κ-carrageenan.

Probing the metabolic phenotype of breast cancer cells by multiple tracer stable isotope resolved metabolomics.

Breast cancers vary by their origin and specific set of genetic lesions, which gives rise to distinct phenotypes and differential response to targeted and untargeted chemotherapies. To explore the functional differences of different breast cell types, we performed Stable Isotope Resolved Metabolomics (SIRM) studies of one primary breast (HMEC) and three breast cancer cells (MCF-7, MDAMB-231, and ZR75-1) having distinct genotypes and growth characteristics, using 13C6-glucose, 13C-1+2-glucose, 13C5,15N2-Gln, 13C3-glycerol, and 13C8-octanoate as tracers. These tracers were designed to probe the central energy producing and anabolic pathways (glycolysis, pentose phosphate pathway, Krebs Cycle, glutaminolysis, nucleotide synthesis and lipid turnover). We found that glycolysis was not associated with the rate of breast cancer cell proliferation, glutaminolysis did not support lipid synthesis in primary breast or breast cancer cells, but was a major contributor to pyrimidine ring synthesis in all cell types; anaplerotic pyruvate carboxylation was activated in breast cancer versus primary cells. We also found that glucose metabolism in individual breast cancer cell lines differed between in vitro cultures and tumor xenografts, but not the metabolic distinctions between cell lines, which may reflect the influence of tumor architecture/microenvironment.

Ginkgo biloba exocarp extracts induces apoptosis in Lewis lung cancer cells involving MAPK signaling pathways.

ETHNOPHARMACOLOGICAL RELEVANCE: A fruit of Ginkgo biloba L. is known as Ginkgo nuts. It is an edible traditional Chinese medicine, and could be used for the treatment of cancer thousands of years ago in China. The extracts prepared from the exocarp of Ginkgo biloba (Ginkgo biloba exocarp extracts, GBEE) has the effects of anti-cancer, immune promotion, anti-aging and etc. AIM OF STUDY: To study the effects of GBEE inducing apoptosis in Lewis lung cancer (LLC) cells and the role of Mitogen-activated protein kinase(MAPK) signaling pathways in it. MATERIALS AND METHODS: The LLC solid tumor model was established in C57BL/6J mice. The tumor-bearing mice were randomly divided into 5 groups. A normal control group without tumor cells was established additionally. There were 10 mice in each group, and they were dosed 24h after inoculation. The GBEE (50, 100, 200mg/kg b.w.) groups were dosed by intragastric gavage (i.g.). The mice in positive control group were intraperitoneal (i.p.) injected with cyclophosphamide (CPA) at a dose of 20mg/kg (b.w.). The model control group and the normal control group were both given normal saline (NS) by i.g.. All the groups were dosed at a volume of 0.1mL/10g (b.w.), once a day for 18d. The day after the last administration, the transplanted tumors was stripped and weighed, and the inhibition rate was calculated. In vitro experiments, MTT method was applied to detect the effects of GBEE on LLC cells and primary cultured mouse lung cells. Annexin V-FITC/PI method was used to detect the apoptosis rate of LLC cells. Rhodamine 123 method was used to detect the Mitochondrial transmembrane potential (MTP). Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to detect the levels of Fas mRNA. Western Blot was used to detect the expression of Bax, Bcl-2, Cyt C, cleaved Caspase-3 and MAPK proteins in the corresponding parts of LLC cells. RESULTS: GBEE (50-200mg/kg) inhibited the growth of LLC transplanted tumors with a dose-effect relationship. GBEE (5-160µg/mL) inhibited the proliferation of LLC cells in vitro with the half maximal inhibitory concentration (IC50) value of 162.43µg/mL, while it had no significant inhibitory effects on the primary cultured mouse lung cells. After GBEE (10, 20 and 40µg/mL) acted on the LLC cells, the apoptosis rate was increased and the MTP was decreased. The ratio of Bax/Bcl-2 was increased in the cells. Meanwhile, it also promoted the translocation of Bax/Bcl-2 in mitochondrial membrane and the release of Cyt C from mitochondria to cytosol. In addition, it up-regulated the cleaved-Caspase-3 protein expression. The mRNA levels of Fas and the protein levels of Fas, FasL and p-p38 in the cells were both increased. The levels of p-ERK1/2 and p-JNK1/2 protein were down-regulated but the p38, ERK1/2 and JNK1/2 were not significantly changed. CONCLUSIONS: GBEE induces apoptosis in LLC cells via mitochondrial-mediated intrinsic pathway and death receptor-mediated extrinsic pathway, which may be closely relevant to the regulation of MAPK signaling pathways.

Formation of κ-carrageenan-gelatin polyelectrolyte complexes studied by (1)H NMR, UV spectroscopy and kinematic viscosity measurements.

The intermolecular interactions between an anionic polysaccharide from the red algae κ-carrageenan and a gelatin polypeptide, forming stoichiometric polysaccharide-polypeptide (bio)polyelectrolyte complexes in the aqueous phase, were examined. The major method of investigation was high-resolution (1)H NMR spectroscopy. Additional data were obtained by UV absorption spectroscopy, light scattering dispersion and capillary viscometry. Experimental data were interpreted in terms of the changing roles of electrostatic interactions, hydrophobic interactions and hydrogen bonds when κ-carrageenan-gelatin complexes are formed. At high temperatures, when biopolymer macromolecules in solution are in the state of random coil, hydrophobic interactions make a major contribution to complex stabilization. At the temperature of gelatin's coil→helix conformational transition and at lower temperatures, electrostatic interactions and hydrogen bonds play a defining role in complex formation. A proposed model of the κ-carrageenan-gelatin complex is discussed.