Diagnosis and genetic testing analysis of limb-girdle muscular dystrophy type 2U caused by a compound heterozygous mutation in the ISPD gene.

Limb-girdle muscular dystrophy (LGMD), a rare group of non-congenital inherited muscle diseases, is characterized by a progressive reduction in muscle tone and force of the proximal limbs. The clinical manifestations and genetic patterns of LGMD are heterogeneous. This study reported on a 10-year-old male patient with LGMD type 2U who experienced muscle weakness in the lower limbs after exercise. Upon admission, the patient's creatine kinase levels were significantly elevated, and hydration and alkalinization therapy were ineffective. Using high-throughput sequencing, muscular dystrophy-related genes were tested in the patient, his parents, and his sister. The patient was found to have a heterozygous deletion of exon 9 of the ISPD gene and a heterozygous missense mutation c.1231C>T (p.Leu411Phe). The patient's father carried the heterozygous missense mutation c.1231C>T (p.Leu411Phe) of the ISPD gene, while his mother and sister carried a heterozygous deletion of exon 9 of the ISPD gene. These mutations have not been reported in existing databases or literature. Conservation and protein structure prediction analyses of the mutation sites indicated that they are highly conserved and located in the C-terminal domain of the ISPD protein, which may affect protein function. Based on the above results and relevant clinical data, the patient was definitively diagnosed with LGMD type 2U. This study enriched the spectrum of ISPD gene mutations by summarizing the patient's clinical characteristics and analyzing new ISPD gene variations. This can aid in the early diagnosis and genetic counseling of the disease.

Synthesis, Characterization of Low Molecular Weight Chitosan Selenium Nanoparticles and Its Effect on DSS-Induced Ulcerative Colitis in Mice.

Selenium nanoparticles have attracted extensive attention due to their good bioavailability and activity. In the present study, a new form of selenium nanoparticle (Low molecular weight chitosan selenium nanoparticles (LCS-SeNPs)) were synthesized in a system of sodium selenite and acetic acid. The size, element state, morphology and elementary composition of LCS-SeNPs were characterized by using various spectroscopic and microscopic measurements. The protection of LCS-SeNPs against dextran sulfate sodium (DSS)-induced intestinal barrier dysfunction and the inherent mechanisms of this process were investigated. The results showed that LCS-SeNPs, with an average diameter of 198 nm, zero-valent and orange-red relatively uniform spherical particles were prepared. LCS-SeNPs were mainly composed of C, N, O and Se elements, of which Se accounted for 39.03% of the four elements C, N, O and Se. LCS-SeNPs reduced colon injury and inflammation symptoms and improved intestinal barrier dysfunction. LCS-SeNPs significantly reduced serum and colonic inflammatory cytokines TNF-α and IL-6 levels. Moreover, LCS-SeNPs remarkably increased antioxidant enzyme GSH-Px levels in serum and colonic tissue. Further studies on inflammatory pathways showed that LCS-SeNPs alleviated DSS-induced colitis through the NF-κB signaling pathway, and relieved inflammatory associated oxidative stress through the Nrf2 signaling pathway. Our findings suggested that LCS-SeNPs are a promising selenium species with potential applications in the treatment of oxidative stress related inflammatory intestinal diseases.

Atmospheric cold plasma: a new approach to modify protein and lipid properties of myofibrillar protein isolate from hairtail (Trichiurus lepturus) fish.

BACKGROUND: Quite recently, considerable attention has been paid to atmospheric cold plasma (ACP) as an eco-friendly and highly efficient technology to modify the functional properties of foods. This study focuses on the effect of ACP on the myofibril protein and lipid quality of hairtail (Trichiurus lepturus) fish. In achieving this, the samples were treated with ACP at 50 kV for different times (30, 60, 120, 180, 240, 300 s). RESULTS: The findings indicated slight changes in peroxide value and thiobarbituric acid reactive substances in the samples treated with ACP. A significant increase (P < 0.05) in the surface hydrophobicity (from 131.71 ± 0.81 µg to 146. 34 ± 0.81 µg), turbidity (from 0.13 ± 0.001 to 0.27 ± 0.01), and water-holding capacity (from 61.63% ± 5.7% to 64.86% ± 1.5%) were detected with treated samples. CONCLUSIONS: We conclude that ACP treatment induces marked changes in the protein and lipid properties of myofibril protein isolated from hairtail fish, which strengthen the gel formation of hairtail fish. © 2021 Society of Chemical Industry.

Combined Effect of the Essential Oil and Collagen Film on the Quality of Pacific Mackerel (Pneumatophorus japonicus) Fillet During Cold Storage.

Essential oils (EOs) and collagen have received recent attention in the seafood industry due to their abilities of antibacterial and seafood preservation individually. However, to the authors' best knowledge, very few publications address the issue of the combined effect of EOs and collagen on seafood preservation. Pacific mackerel is one of the most economically valuable fish species in China and easy to deteriorate during storage. Therefore, present study investigated the effect of combined EOs (cinnamon, oregano, and clove) and collagen on the quality of Pacific mackerel during cold storage. A suite of microbiological, physical, and chemical properties that are indicative of quality was measured. From the results, mackerel fillets treated with an EO-collagen film had a smaller increase in microbial counts compared with control. Furthermore, total volatile basic nitrogen (TVB-N), thiobarbituric acid related substance, and pH of mackerel fillet were lower when treated with an EO-collagen film and somewhat lower when treated with collagen alone. According to texture measurements of muscle, samples treated with EO-collagen film began to deteriorate in 8 d, versus only 4 d for control samples. EOs likely contributed to antibacterial and antioxidative activity, and the collagen film isolated muscle from air, which in turn reduced oxidation and retained the quality. Consequently, combination of EOs and collagen film efficiently extends shelf-life of Pacific mackerel during storage.

Bitter peptides from enzymatically hydrolyzed protein increase the number of leucocytes and lysozyme activity of large yellow croaker (Larimichthys crocea).

The large yellow croakers (Larimichthys crocea) are mainly present in the Chinese coast and near seas with high economic importance, but vulnerable to many diseases, especially in the breeding and aquaculture. The purpose of this research was to boost the innate immune system of the large yellow croaker by administering bitter peptides into their peritoneal cavity. Total 120 Juvenile of large yellow croakers in very even weight of 60 g were divided into 4 different groups in 200/300 L of water tank, respectively. Fish growth were observed for 3 months before and after different treatments. The bitter peptides from pepsin hydrolysis were applied because they possess the highest bitter sensory scores. The blood of fish from the different groups was collected and tested for different immune parameters to evaluate the effectiveness of bitter peptides as immune stimulants after administration for 8 weeks. The average ratio of leukocytes/total blood cells (%) for control was found at 14.6%, for the low dose of bitter peptides 0.6 mg/fish was at 29.3%, for middle dose of 1.2 mg/fish was at 35%, and high dose of 2.4 mg/fish was at 30%. The lysozyme assay showed that the OD (optical density) units of relative progress lysis activity at 60 min were 0.17, 0.101, 0.307 and 0.198, respectively. Similarly in the same order as in phagocyte assay, most importantly the middle dose (1.2mg/fish) gave the highest survival rate throughout the assay. The results showed that bitter peptides can be used as immune boosters for the yellow croakers and the optimum dose was 1.2 mg/fish due to both leukocytes and lysozyme activity in the treated samples increased significantly compared with the control group. According to the results obtained, we suggest that the incorporation of middle dose of bitter peptides into fish feeds may reduce the fish diseases in aquaculture, at least for large yellow croakers.

Potential Utilization of Green Tea Leaves and Fenugreek Seeds Extracts as Natural Preservatives for Pacific White Shrimp During Refrigerated Storage.

Ethanolic extracts of green tea leaves (GTE) and fenugreek seeds (FSE) were screened for their antibacterial activity against four food pathogenic strains using disc diffusion method. The two extracts revealed antimicrobial activity against selected bacterial strains. GTE showed the highest antibacterial activity to Escherichia coli and Staphylococcus aureus at a concentration of 1% with inhibition zone equal to 29.45 ± 0.64 mm and 25.68 ± 1.2 mm, respectively. In addition, the effect of GTE and FSE combined with chitosan coating on the shelf life of Pacific white shrimp (PWS) (Litopenaeus vannamei) during refrigerated storage have been studied. Our results indicated that using GTE or FSE during the refrigerated storage of PWS led to significantly decreased Total Volatile Bases Nitrogen, Thiobarbituric acid reacting substances, total bacterial count, and pH. The sensory properties of PWS have improved considerably in the samples treated GTE or FSE. These findings suggested that the application of chitosan coating combined with GTE or FSE to PWS is advisable to achieve better quality during refrigerated storage.

Protease Hydrolysates of Filefish (Thamnaconus modestus) Byproducts Effectively Inhibit Foodborne Pathogens.

In this study, novel antimicrobial peptides had been derived by enzymatic hydrolysis of filefish (Thamnaconus modestus) byproduct (HFBP). Different proteases, (papain [P], trypsin [T], neutrase [N], pepsin [PE], and the mixture I [PN] and mixture II [PT]) and different hydrolysis time (60, 120, 180, and 240 min), have been used to generate peptides with different lengths and amino acid sequences. The antimicrobial properties of HFBPs were tested, against Gram-positive and Gram-negative strains, using disc diffusion method. HFBP prepared after 120 min of the enzymatic hydrolysis by trypsin (HFBP-T) exhibited greatest antibacterial activities. Bacillus cereus 10451 (BC) and Salmonella enteritidis 10982 (SE) strains were most sensitive to HFBP-T with an inhibitory zone of 24.68 and 29.67 mm diameter and minimum inhibitory concentration of 1.25 and 2.5 mg/mL, respectively. Moreover, the antimicrobial activities of tested HFBPs increased significantly at low pH and temperature. The amino acid analysis showed that HFBP-T protein hydrolysate is high in an amino acid of proline, which probably contributes to the antimicrobial activity. The results obtained from scanning electron microscopy suggested that HFBPs might kill bacteria by acting on the cell wall of bacteria. Conclusively, the HFBP derived from filefish byproduct with biological activates is an interesting alternative to the use of waste from the fishing industry as natural antimicrobials in food stuff.

Chemical compositions and nutritional profiles of two edible tunicate species (Halocynthia roretzi and Halocynthia aurantium).

As an abundant marine bioresource, tunicates could be exploited in the food industry. However, limited knowledge of their chemical composition and nutritional profiles prohibited further application. In this study, two common edible tunicate species, Halocynthia roretzi (HR) and Halocynthia aurantium (HA), were subjected to comprehensive composition analysis in terms of moisture, protein, lipids, cellulose, ash, amino acids, fatty acids, non-cellulose carbohydrates and minerals. Reddish HR was much bigger than purple HA with respect to body length and weight, and their moisture fell within 82.98 %-90.92 %. The non-edible outer shell part (OS) and edible internal organs part (IO) had a dry weight ratio of around 3:2 for both two species. Generally, for both HR and HA, IO was more abundant in protein and lipids. In contrast, OS had much higher cellulose contents, confirming the better suitability of IO as a nutritional seafood. IO was richer in essential amino acids and unsaturated fatty acids, while OS had more abundant saturated fatty acids. The detected non-cellulose monosugars ranged from 0.47 % to 1.18 % and indicated the presence of some sulfated glycans. IO of HR had higher contents of essential minerals, such as Cu, Zn, and Fe, while IO of HA showed a higher K content. To sum up, this study identified the chemical composition and nutritional profile variations among different tunicate species and various dissected parts, guiding the development of specific strategies to exploit tunicates for proper food applications.

Cellulose-based fluorescent materials fabricated in CO2 switchable solvent for freshness monitoring.

Herein, a novel cellulose-based fluorescent material with amine-responsive property was developed in a CO2 switchable solvent for the contactless, real-time, and visual monitoring of the freshness of shrimp, crab, and fish. A nucleophilic cellulose carbonate intermediate is in-situ formed upon dissolution of cellulose in the CO2 switchable solvent, which is adopted for the highly effective grafting of poly(l-lactic acid) and amine-responsive fluorescent reagent 7-amino-4-(trifluoromethyl) coumarin onto the cellulose backbone via ring opening polymerization and nucleophilic substitution reaction, respectively. The product has good processibility, and sensitive and reversible fluorescent responsive towards ammonia. The fluorescent intensity of the material decreased significantly and its fluorescent color changed from blue to cyan, when the aquatic products got spoiled. This study indicates that the CO2 switchable solvent has great potential for the modification of cellulose with various functionality by taking advantage of the highly active nucleophilic cellulose carbonate intermediate.

Preparation, identification, activity prediction, and protective effects on IR-HepG2 cells of five novel DPP-IV inhibitory peptides from protein hydrolysate of skipjack tuna dark muscles.

To produce peptides with high dipeptidyl peptidase IV (DPP-IV) inhibitory activity, neutrase was selected from five proteases (trypsin, neutrase, pepsin, alcalase and flavor protease) with the highest degree of hydrolysis (DH) (18.23 ± 1.08%) and DPP-IV inhibitory rate (53.35 ± 4.02%) to produce protein hydrolysate (NPH) from the dark muscles of skipjack tuna (Katsuwonus pelamis). Then, NPH-1 was isolated from NPH by gel permeation chromatography and found to possess the highest DPP-IV inhibitory rate (65.12 ± 7.94% at 0.5 mg ml-1) in the separated components (including NPH-1, NPH-2, NPH-3 and NPH-4). Subsequently, the available prediction models of tripeptides and tetrapeptides with the DPP-IV inhibitory rate were established using an artificial neural network (ANN). The RMSE (0.56 and 0.33 for the model established through collected tripeptides and tetrapeptides, respectively) and R2 (0.95 and 0.99 for the model established through collected tripeptides and tetrapeptides, respectively) of the ANN model's parameters were within acceptable limits, indicating that this model is available. Next, the ANN model was applied to predict tripeptides and tetrapeptides from the hydrolysate of skipjack tuna dark muscles, and five peptides (Ala-Pro-Pro (APP), Pro-Pro-Pro (PPP), Asp-Pro-Leu-Leu (DPLL), Glu-Ala-Val-Pro (EAVP) and Glu-Ala-Iie-Pro (EAIP)) possessing a noticeable DPP-IV inhibitory rate (with DPP-IV IC50 values of 42.46 ± 5.02, 37.71 ± 9.17, 58.85 ± 14.42, 49.94 ± 6.69 and 57.15 ± 6.13 µM, respectively) were screened from the protein hydrolysate. The above five peptides were proved to effectively promote glucose consumption in the insulin resistant-HepG2 (IR-HepG2) cell model considering that the glucose consumption rates of APP, PPP, DPLL, EAVP and EAIP treatment groups are all more than twice that of the dexamethasone group. Accordingly, mechanistic studies showed that these peptides interacted with PI3K/AKT and AMPK signaling pathways and promoted the phosphorylation of PI3K p110, AKT and AMPK (the protein expressions of PI3K p110, p-AKT and p-AMPK in APP, PPP, DPLL, EAVP and EAIP treatment groups are 1.64-2.22 fold compared with that in the dexamethasone group), thereby enhancing glucose uptake and further alleviating insulin resistance. These findings demonstrated that skipjack tuna dark muscle is a potential DPP-IV inhibitory peptide source, and five DPP-IV inhibitory peptides from its hydrolysate may exert potent anti-diabetic activity. In comparison, PPP may be the most potential active ingredient for healthy food against type 2 diabetes mellitus in the five screened peptides considering synthetically the DPP-IV inhibitory rate, bioavailability and synthesis cost.

Emulsions containing composite (clove, oregano, and cinnamon) essential oils: Phase inversion preparation, physicochemical properties and antibacterial mechanism.

Natural essential oils (EOs), especially those combining different individual EOs (also termed composite EOs) with enhanced performance, are becoming healthy, market-sought food preservatives/additives. This study aims to provide insights into the challenge regarding EOs processing due to their low solubility and the elusive mechanism under the enhanced bio-reactivity of composite EOs. A unique oil/water interacting network was created by phase-inversion processing, which enhances EO solubilization and emulsification to form composite EO formulations (EOFs) containing ordinary cinnamon, oregano and clove EOs. These EOFs mainly contained cinnamaldehyde, carvacrol and eugenol and exhibited excellent post-storage stability. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging ability of EOFs (at 15.880 µL/mL) was > 88%, and the Ferric reducing antioxidant power (FRAP) was 1.8 mM FeSO4·7H2O. The minimum inhibitory concentration (MIC) of EOFs against E. coli and S. aureus was ∼7.940 µL/mL. The EOFs could cause quick deterioration of bacterial structures, demonstrating high efficacy in bacteria-killing and anti-biofilm formation.

Efficacy of Chitosan Oligosaccharide Combined with Cold Atmospheric Plasma for Controlling Quality Deterioration and Spoilage Bacterial Growth of Chilled Pacific White Shrimp (Litopenaeus vannamei).

A novel food processing technique based on the combination of cold atmospheric plasma (CAP) and chitosan oligosaccharide treatment (COS) was developed to enhance antibacterial performance and extend the shelf life of Pacific white shrimp (Litopenaeus vannamei). Effects of different treatments on the microbial community composition, physicochemical properties, and post-storage behaviors of Pacific white shrimp were evaluated during chilled storage for up to 10 days. Results showed that the synergistic effects of COS and CAP could be obtained, largely inhibiting the growth of microorganisms. The content of total volatile basic nitrogen (TVB-N), total viable counts (TVC), and pH value in treated groups were lower than in the control group and the loss of moisture content, water activity, and sensory score were observed. Compared to the control group, shrimp was on the verge of spoilage on the 6th day of storage, while the COS-CAP-treated shrimp had a 4-day lag period. Moreover, the COS and CAP could effectively inhibit the growth of Aliivibrio, the predominant microbial group in the ultimate storage period. This study suggests that the combined utilization of COS and CAP could be a high-efficacy technique for extending the shelf-life of shrimp.

Assessment of the Effect of Cold Atmospheric Plasma (CAP) on the Hairtail (Trichiurus lepturus) Quality under Cold Storage Conditions.

Cold Atmospheric Plasma (CAP) is a novel non-thermal preservation method that extends the shelf-life of food. Therefore, this study investigated the effect of CAP on the quality parameters of hairtail (Trichiurus lepturus) during cold storage conditions (at 4 °C and RH range 45−55%). For that reason, different quality parameters including the total bacteria count (TBC), total volatile basic nitrogen (TVB-N), pH, thiobarbituric acid reacting substances value (TBARS), color, texture, and sensory evaluation have been measured. The hairtail was exposed to CAP at 50 kV voltage for 2, 3, 4, and 5 min. The results showed that the samples treated with CAP at 50 kV for 5 min had significantly lower (p < 0.05) TBC (7.04 ± 0.26 log CFU/g) compared with the control sample (8.69 ± 0.06 log CFU/g). Similar results were found concerning TVB-N, which strongly decreased in the treated samples (16.63 ± 0.03 mg N/100 g) in comparison with the control sample (22.79 ± 0.03 mg N/100 g). In addition, the CAP-treated samples had lower (p < 0.05) changes in color than those of the control group. With reference to the sensory evaluation, the shelf-life of CAP-treated samples (at 50 kV for 5 min) was longer than the untreated samples by about 6 days. These results led us to the conclusion that CAP can effectively delay spoilage and deterioration, slow the rise in pH, and maintain the sensory attributes of hairtail during cold storage conditions.

Changes in Biochemical Properties and Activity of Trypsin-like Protease (Litopenaeus vannamei) Treated by Atmospheric Cold Plasma (ACP).

The changes in the functional properties of trypsin from shrimps (Litopenaeus vannamei) after, Atmospheric Cold Plasma (ACP) treatments, have been evaluated in terms of enzyme inactivation, surface hydrophobicity, secondary structure, fluorescence intensity, and particle size distribution. Different exposure voltages of 10, 20, 30, 40, and 50 kV at various treatment times (1, 2, 3, and 4 min) have been employed, in a separate assay. The results showed that trypsin-like protease activity decreased (by about 50%), and the kinetic constants Km value increased, while the kcat value decreased. Surface hydrophobicity and fluorescence intensity revealed a significant increase compared to the control sample. A high degree of protein degradation has been noticed by SDS-PAGE analysis. In addition, circular dichroism indicated that random coil and α-helix contents declined while ß-turn and ß-sheet contents have raised. A sharp drop in the particle size was observed with increasing the treatment voltage from 0 to 40 kV for 4 min, and the corresponding peak reached the minimum of 531.2 nm. Summing up the results, it can be concluded that the ACP technique effectively affects the activity of trypsin-like protease, which in terms enhances the quality of dietary protein.

Understanding the role of atmospheric cold plasma (ACP) in maintaining the quality of hairtail (Trichiurus Lepturus).

Impacts of atmospheric cold plasma (ACP) on the properties of muscle protein and performance of extracted crude enzyme of hairtail (Trichiurus Lepturus) fish have been evaluated. A decrease in extracted crude enzyme activity with increasing the ACP treatment time has been found, and the highest reduction (p < 0.05) value of 0.035 units/mg proteins was obtained after 240 s. A considerable increase in the carbonyl content in the treated sample for about three times higher than the control sample was found, and a decrease of total sulfhydryl content to 0.34 nmol/mg protein. Texture profile analysis, water holding capacity, and the color properties of the muscle protein improved significantly in the samples treated with ACP. SDS-PAGE pattern showed an increase in the band intensity of cross-linked myosin heavy chains and actin proteins. Based on these outcomes, ACP could play a significant role as a promising non-thermal method to prolong the shelf-life of hairtail fish.

Effects of cold atmospheric plasma on squid proteases and gel properties of protein concentrate from squid (Argentinus ilex) mantle.

The effect of cold atmospheric plasma (CAP) on protein concentrate from squid (Argentinus ilex) mantle has been evaluated in terms of gel properties, protease inhibition, texture profile, color attributes, and water holding capacity. Different exposure time (15, 60, 120, 180, 240 and 300 s) at 60 kV have been employed. Our results indicated that protease activity decreases with increasing treatment time. The highest reduction (p < 0.05) in protease activity (64%) was observed after 240 s of CAP treatment. Texture profile analysis, color properties and water holding capacity of the treated squid gel revealed a significant increase. Protein carbonyl and sulfhydryl group contents findings' showed a significant increase in carbonyl content (about three times of the original content), while the total sulfhydryl group decreased (up to about 40%) in the crude extract. Microstructure and SDS-PAGE analysis revealed a high degree of protein aggregation in the squid gel treated with CAP.

Physicochemical changes of myofibrillar proteins of squid (Argentinus ilex) induced by hydroxyl radical generating system.

The effect of a hydroxyl radical generating system (HRGS), which contained FeCl3, sodium ascorbate, and different concentrations of H2O2, on the physiochemical properties of myofibrillar protein (MP) from squid mantles, has been investigated. The effect of different exposure times to HRGS was also considered. Compared to non-oxidized MP, a significant (p < 0.05) increase in carbonyl content (more than 50% of its original content) and protein solubility, as well as in surface hydrophobicity, was observed in the oxidative MP. With different treatment times, a sharp decrease (p < 0.05) in sulfhydryl content was detected. In addition, hydroxyl radical treatment significantly reduced the MP gel's texture properties, whiteness and water holding capacity, especially at higher concentrations of H2O2. This observation could be attributed to extensive disorderly and less compact structure of MP gels. The results demonstrate the negative effect of HRGS on the structural and functional properties of MP from squid mantles.

Antimicrobial Properties and Mechanism of Action of Some Plant Extracts Against Food Pathogens and Spoilage Microorganisms.

This work aims to evaluate the antimicrobial potential of ethanolic and water extracts of roselle (Hibiscus sabdariffa), rosemary (Rosmarinus officinalis), clove (Syzygium aromaticum), and thyme (Thymus vulgaris) on some food pathogens and spoilage microorganisms. Agar well diffusion method has been used to determine the antimicrobial activities and minimum inhibitory concentrations (MIC) of different plant extracts against Gram-positive bacteria (Bacillus cereus, Staphylococcus aureus), Gram-negative bacteria (Escherichia coli, Salmonella enteritidis, Vibrio parahaemolyticus, and Pseudomonas aeruginosa), and one fungus (Candida albicans). The extracts exhibited both antibacterial and antifungal activities against tested microorganisms. Ethanolic roselle extract showed significant antibacterial activity (P < 0.05) against all tested bacterial strains, while no inhibitory effect on Candida albicans (CA) was observed. Only the ethanolic extracts of clove and thyme showed antifungal effects against CA with inhibition zones ranging from 25.2 ± 1.4 to 15.8 ± 1.2 mm, respectively. Bacillus cereus (BC) appears to be the most sensitive strain to the aqueous extract of clove with a MIC of 0.315%. To enhance our understanding of antimicrobial activity mechanism of plant extracts, the changes in internal pH (pHint), and membrane potential were measured in Staphylococcus aureus (SA) and Escherichia coli (EC) cells after exposure to the plant extracts. The results indicated that the plant extracts significantly affected the cell membrane of Gram-positive and Gram-negative bacteria, as demonstrated by the decline in pHint as well as cell membrane hyperpolarization. In conclusion, plant extracts are of great value as natural antimicrobials and can use safely as food preservatives.

Inhibition of Migration and Invasion of Hepatocarcinoma HepG2 Cells by Dietary Saponins via Targeting HIF-1α.

BACKGROUND: Different saponins from herbs have been used as tonic or functional foods, and for treatment of various diseases including cancers. Although clinical data has supported the function of these saponins, their underlying molecular mechanisms have not been well defined. METHODS: With the simulated hypoxia created by 8 hours of Cu++ exposure and following 24 hour incubation with different concentration of saponins in HepG2 cells for MTT assay, migration and invasion assays, and for RT-PCR, and with each group of cells for immunofluorescence observation by confocal microscopy. Results：ZC-4 had the highest rate of inhibition of cell proliferation by MTT assay, and the highest inhibition of migration rate by in vitro scratch assay, while ZC-3 had the highest inhibition of invasion ratio by transwell assay. Under the same simulated hypoxia, the molecular mechanism of saponin function was conducted by measuring the gene expression of Hypoxia Inducible Factor (HIF)-1α through RT-PCR, in which ZC-3 showed a potent inhibition of gene HIF-1α. For the protein expression by immunofluorescence staining with confocal microscopy, HIF-1α was also inhibited by saponins, with the most potent one being ZC-4 after eight hours' relatively hypoxia incubation. CONCLUSION: Saponins ZC-4 and ZC-3 have the potential to reduce HepG2 cell proliferation, migration and invasion caused by hypoxia through effectively inhibiting the gene and protein expression of HIF-1α directly and as antioxidant indirectly.

Characterization of polyphenol oxidase from litchi pericarp using (-)-epicatechin as substrate.

Polyphenol oxidase (PPO) from litchi (Litchi chinensis Sonn.) pericarp was characterized using (-)-epicatechin, which was the major endogenous polyphenol in litchi pericarp as a substrate. The optimum pH for PPO activity with (-)-epicatechin was 7.5, and the enzyme was unstable below pH 4.5 and stable in the pH range of 6.0-8.0. Residual activities of PPO were 86.25, 86.31, and 80.17% after 67 days of incubation at 4 degrees C at pH 6.0, 7.5, and 8.0, respectively. From thermostability studies, the Ki value increased with temperature and the results suggested that the enzyme was unstable above 45 degrees C. Moreover, the results also provided strong evidence that the denaturalization temperature of PPO was near 70 degrees C. The inhibition studies indicated that l-cysteine and glutathione were strong inhibitors even at low concentrations while NaF inhibited moderately. In addition, the results also indicated that the inhibition mechanisms of thiol groups were different from those of halide salts.