Redundant role of OsCNGC4 and OsCNGC5 encoding cyclic nucleotide-gated channels in rice pollen germination and tube growth.

In staple crops, such as rice (Oryza sativa L.), pollen plays a crucial role in seed production. However, the molecular mechanisms underlying rice pollen germination and tube growth remain underexplored. Notably, we recently uncovered the redundant expression and mutual interaction of two rice genes encoding cyclic nucleotide-gated channels (CNGCs), OsCNGC4 and OsCNGC5, in mature pollen. Building on these findings, the current study focused on clarifying the functional roles of these two genes in pollen germination and tube growth. To overcome functional redundancy, we produced gene-edited rice plants with mutations in both genes using the CRISPR-Cas9 system. The resulting homozygous OsCNGC4 and OsCNGC5 gene-edited mutants (oscngc4/5) exhibited significantly lower pollen germination rates than the wild type (WT), along with severely reduced fertility. Transcriptome analysis of the double oscngc4/5 mutant revealed downregulation of genes related to receptor kinases, transporters, and cell wall metabolism. To identify the direct regulators of OsCNGC4, which form a heterodimer with OsCNGC5, we screened a yeast two-hybrid library containing rice cDNAs from mature anthers. Subsequently, we identified two calmodulin isoforms (CaM1-1 and CaM1-2), NETWORKED 2 A (NET2A), and proline-rich extension-like receptor kinase 13 (PERK13) proteins as interactors of OsCNGC4, suggesting its roles in regulating Ca2+ channel activity and F-actin organization. Overall, our results suggest that OsCNGC4 and OsCNGC5 may play critical roles in pollen germination and elongation by regulating the Ca2+ gradient in growing pollen tubes.

Rice pollen-specific OsRALF17 and OsRALF19 are essential for pollen tube growth.

Pollen tube growth is essential for successful double fertilization, which is critical for grain yield in crop plants. Rapid alkalinization factors (RALFs) function as ligands for signal transduction during fertilization. However, functional studies on RALF in monocot plants are lacking. Herein, we functionally characterized two pollen-specific RALFs in rice (Oryza sativa) using multiple clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated protein 9-induced loss-of-function mutants, peptide treatment, expression analyses, and tag reporter lines. Among the 41 RALF members in rice, OsRALF17 was specifically expressed at the highest level in pollen and pollen tubes. Exogenously applied OsRALF17 or OsRALF19 peptide inhibited pollen tube germination and elongation at high concentrations but enhanced tube elongation at low concentrations, indicating growth regulation. Double mutants of OsRALF17 and OsRALF19 (ralf17/19) exhibited almost full male sterility with defects in pollen hydration, germination, and tube elongation, which was partially recovered by exogenous treatment with OsRALF17 peptide. This study revealed that two partially functionally redundant OsRALF17 and OsRALF19 bind to Oryza sativa male-gene transfer defective 2 (OsMTD2) and transmit reactive oxygen species signals for pollen tube germination and integrity maintenance in rice. Transcriptomic analysis confirmed their common downstream genes, in osmtd2 and ralf17/19. This study provides new insights into the role of RALF, expanding our knowledge of the biological role of RALF in regulating rice fertilization.

Cinnamomum cassia and Rosa laevigata Mixture Improves Benign Prostatic Hyperplasia in Rats by Regulating Androgen Receptor Signaling and Apoptosis.

Benign prostatic hyperplasia (BPH) is the most common condition in elderly men that is characterized by an increase in the size of the prostate gland. Cinnamomum cassia and Rosa laevigata have been reported to treat the symptoms associated with BPH. The aim of this study was to evaluate the effects of HT080, an herbal extract of C. cassia and R. laevigata, on a testosterone propionate (TP)-induced BPH rat model. The rats received a daily subcutaneous injection of TP (3 mg/kg) for 4 weeks to induce BPH. Rats were divided into four groups: group 1 (sham), group 2 (BPH, TP alone), group 3 (Fina, TP + finasteride 1 mg/kg/day), and group 4 (HT080, TP + HT080 200 mg/kg/day). At the end of the experiment, all rats were sacrificed, and their prostate glands were removed, weighed, and subjected to histopathological examination and western blot analyses. Serum testosterone and dihydrotestosterone (DHT) levels were determined. In addition, serum alanine and aspartate aminotransferase levels were measured to evaluate the toxicity in the liver. The Hershberger bioassay was also conducted to investigate the effects of HT080 on androgenic and antiandrogenic activities. In the BPH model, the prostate weight, prostate index, prostate epithelial thickness, and serum testosterone and DHT levels in the HT080 group were significantly reduced compared to the BPH group. Histological studies showed that HT080 reduced prostatic hyperplasia. The protein expression of androgen receptor from the HT080 group was significantly reduced in comparison with the BPH group (p < 0.05). HT080 also induced apoptosis by regulating Bcl-2 and Bax expression. In addition, HT080 showed no toxicity in the liver and did not exhibit androgenic and antiandrogenic activities. Our finding revealed that HT080 can be a potential candidate for the treatment of BPH by regulating androgen receptor signaling and apoptosis.

Cell-Free Supernatant of Bacillus thuringiensis Displays Anti-Biofilm Activity Against Staphylococcus aureus.

Staphylococcus aureus is an important bacterial pathogen responsible for biofilm formation in medical devices. Due to the increasing antibiotic resistance of S. aureus, it is necessary to search for new anti-biofilm agents. In this study, the cell-free supernatant of Bacillus thuringiensis inhibited biofilm formation up to 93% and dispersed biofilms up to 83% without affecting the growth of S. aureus. The ethyl acetate extract of B. thuringiensis cell-free supernatant exhibited a dose-dependent anti-biofilm activity against S. aureus with the biofilm inhibition concentration ranging from 8 to 64 µg/mL. Scanning electron microscopy revealed that the cell-free supernatant extract of B. thuringiensis resulted in a significant reduction in S. aureus biofilms. The ethyl acetate extract of cell-free supernatant of B. thuringiensis was found to contain various compounds with structural similarity to known anti-biofilm compounds. In particular, squalene, cinnamic acid derivatives, and eicosapentaene seem to act synergistically against S. aureus biofilms. Hence, B. thuringiensis cell-free supernatant proved to be effective against S. aureus biofilms. The results clearly show the potential of natural molecules produced by B. thuringiensis as alternative therapies with anti-biofilm activity instead of bactericidal properties.

Correlation between the microbiome and pack burst spoilage of Allium sativum supplemented fermented hot pepper paste.

Gochujang (fermented hot pepper paste) products are well known for their distinct, spicy flavor. However, frequent pack burst spoilage of gochujang products occurs during transportation and storage because of microbial aerogenesis, resulting in considerable economic losses. The present study aimed to prevent pack burst spoilage of gochujang products by supplementing them with garlic ethanol extract. A simulated pack burst experiment revealed that 42.86 % of normal gochujang products were spoiled. Garlic ethanol extract significantly inhibited the growth of Zygosaccharomyces rouxii in gochujang products, with low minimum inhibitory concentration values (12.5-25 mg/mL). Gochujang products supplemented with various concentrations (1 % and 2.5 %) of garlic ethanol extract exhibited marked inhibition of microbial growth, particularly Z. rouxii, and pack burst spoilage. Microbiome analysis revealed that the pack burst samples harbored a high abundance of Z. rouxii. Supplementation of gochujang with 1 % garlic ethanol extract drastically reduced Z. rouxii abundance and prevented pack burst. Moreover, gochujang products supplemented with 1 % garlic ethanol extract exhibited a high hedonic score in the sensory analysis. Based on the results of this study, we concluded that supplementation of gochujang products with 1 % garlic ethanol extract before packaging could be effective in preventing pack burst spoilage of gochujang.

Can Low-Dose of Dietary Vitamin E Supplementation Reduce Exercise-Induced Muscle Damage and Oxidative Stress? A Meta-Analysis of Randomized Controlled Trials.

Vitamin E plays an important role in attenuating muscle damage caused by oxidative stress and inflammation. Despites of beneficial effects from antioxidant supplementation, effects of antioxidants on exercise-induced muscle damage are still unclear. The aim of this meta-analysis was to investigate the effects of dietary vitamin E supplementation on exercise-induced muscle damage, oxidative stress, and inflammation in randomized controlled trials (RCTs). The literature search was conducted through PubMed, Medline, Science Direct, Scopus, SPORTDiscuss, EBSCO, Google Scholar database up to February 2022. A total of 44 RCTs were selected, quality was assessed according to the Cochrane collaboration risk of bias tool (CCRBT), and they were analyzed by Revman 5.3. Dietary vitamin E supplementation had a protective effect on muscle damage represented by creatine kinase (CK; SMD -1.00, 95% CI: -1.95, -0.06) and lactate dehydrogenase (SMD -1.80, 95% CI: -3.21, -0.39). Muscle damage was more reduced when CK was measured immediately after exercise (SMD -1.89, 95% CI: -3.39, -0.39) and subjects were athletes (SMD -5.15, 95% CI: -9.92, -0.39). Especially vitamin E supplementation lower than 500 IU had more beneficial effects on exercise-induced muscle damage as measured by CK (SMD -1.94, 95% CI: -2.99, -0.89). In conclusion, dietary vitamin E supplementation lower than 500 IU could prevent exercise-induced muscle damage and had greater impact on athletes.

Insights into cyclooxygenase-2 inhibition by isolated bioactive compounds 3-caffeoyl-4-dihydrocaffeoyl quinic acid and isorhamnetin 3-O-ß-D-glucopyranoside from Salicornia herbacea.

BACKGROUND: Cyclooxygenase-2 (COX-2) is an important enzyme with numerous biological functions. Overexpression of COX-2 has been associated with various inflammatory-related diseases and therefore, projected as an important pharmacological target. PURPOSE: We aimed to investigate the inhibitory potential of isolated bioactive compounds, 3-caffeoyl-4-dihydrocaffeoyl quinic acid (CDQ) and isorhamnetin 3-O-ß-d-glucopyranoside (IDG), from Salicornia herbacea against COX-2 using both computational and in vitro approaches. METHODS: Computational analysis, including molecular docking, molecular dynamics (MD) simulations, and post-simulations analysis, were employed to estimate the binding affinity and stability of CDQ and IDG in the catalytic pocket of COX-2 against Celecoxib as positive control. These predictions were further evaluated using in vitro enzyme inhibition as well as gene expression mediation in macrophages cells. RESULTS: Molecular docking analysis revealed substantial binding energy of CDQ (-6.1 kcal/mol) and IDG (-5.9 kcal/mol) with COX-2, which are lower than Celecoxib (-8.1 kcal/mol). MD simulations (100 ns) and post simulation analysis exhibited the substantial stability and binding affinity of docked CDQ and IDG compounds with COX-2. In vitro assays indicated significant COX-2 inhibition by CDQ (IC50 = 76.91 ± 2.33 µM) and IDG (IC50 = 126.06 ± 9.44 µM). This result supported the inhibitory potential of isolated bioactive compounds against COX-2. Also, a cellular level study revealed a downregulation of COX-2 expression in tumor necrosis factor-alpha stimulated RAW 264.7 macrophages treated with CDQ and IDG. CONCLUSION: Computational and experimental analysis of CDQ and IDG from S. herbacea established their potential in the inhibition and mediation of COX-2. Hence, CDQ and IDG can be considered for therapeutic development against COX-2 linked disorders, such as inflammation and cancer. Furthermore, CDQ and IDG structures can be served as a lead compound for the development of advanced novel anti-inflammatory drugs.

OsMTD2-mediated reactive oxygen species (ROS) balance is essential for intact pollen-tube elongation in rice.

The highly specialized haploid male gametophyte-pollen consist of two sperm cells and a large vegetative cell. Successful fertilization requires proper growth timing and rupture of the pollen tube until it delivers sperm cells, which occur immediately after a pollen grain hydrates. Although a tight regulation on polar cell-wall expansion of the pollen tube is fundamentally important, the underlying molecular mechanism remains largely unknown, especially in crop plants. Here, we characterized the function of male-gene transfer defective 2 (OsMTD2) gene in rice (Oryza sativa), which belongs to the plant-specific receptor-like kinase, the CrRLK1L family. We demonstrated that OsMTD2 is an essential male factor participating in pollen-tube elongation based on genetic evidence and physiological observations. Because of unavailability of homozygous mutant via conventional methods, we used CRISPR-Cas9 system to obtain homozygous knockout mutant of OsMTD2. We were able to identify phenotypic changes including male sterility due to early pollen-tube rupture in the mutant. We observed that the production of reactive oxygen species (ROS) was dramatically reduced in mutants of OsMTD2 pollen grain and tubes with defective pectin distribution. Transcriptome analysis of osmtd2-2 versus wild-type anthers revealed that genes involved in defense responses, metabolic alteration, transcriptional and protein modification were highly upregulated in the osmtd2-2 mutant. Through yeast-two-hybrid screening, we found that OsMTD2 kinase interacts with E3 ligase SPL11. Taken together, we propose that OsMTD2 has crucial functions in promoting pollen-tube elongation through cell-wall modification, possibly by modulating ROS homeostasis during pollen-tube growth.

Analysis of sticky generative cell mutants reveals that suppression of callose deposition in the generative cell is necessary for generative cell internalization and differentiation in Arabidopsis.

In flowering plants, double fertilization between male and female gametophytes, which are separated by distance, largely depends on the unique pattern of the male gametophyte (pollen): two non-motile sperm cells suspended within a tube-producing vegetative cell. A morphological screen to elucidate the genetic control governing the strategic patterning of pollen has led to the isolation of a sticky generative cell (sgc) mutant that dehisces abnormal pollen with the generative cell immobilized at the pollen wall. Analyses revealed that the sgc mutation is specifically detrimental to pollen development, causing ectopic callose deposition that impedes the timely internalization and differentiation of the generative cell. We found that the SGC gene encodes the highly conserved domain of unknown function 707 (DUF707) gene that is broadly expressed but is germline specific during pollen development. Additionally, transgenic plants co-expressing fluorescently fused SGC protein and known organelle markers showed that SGC localizes in the endoplasmic reticulum, Golgi apparatus and vacuoles in pollen. A yeast two-hybrid screen with an SGC bait identified a thaumatin-like protein that we named GCTLP1, some homologs of which bind and/or digest ß-1,3-glucans, the main constituent of callose. GCTLP1 is expressed in a germline-specific manner and colocalizes with SGC during pollen development, indicating that GCTLP1 is a putative SGC interactor. Collectively, our results show that SGC suppresses callose deposition in the nascent generative cell, thereby allowing the generative cell to fully internalize into the vegetative cell and correctly differentiate as the germline progenitor, with the potential involvement of the GCTLP1 protein, during pollen development in Arabidopsis.

Glycosmis pentaphylla (Rutaceae): A Natural Candidate for the Isolation of Potential Bioactive Arborine and Skimmianine Compounds for Controlling Multidrug-Resistant Staphylococcus aureus.

Several multidrug-resistant organisms have emerged, which increases the threat to public health around the world and a limited number of therapeutics were available to counteract these issues. Thus, researchers are trying to find out novel antimicrobials to overcome multidrug-resistant issues. The present study aimed to isolate antibacterial principles against the clinical isolates of multidrug-resistant (MDR) Staphylococcus aureus from the ethyl acetate extract of Gycosmis pentaphylla. The isolation and structural characterization of bioactive compounds were carried out using various chromatographic techniques (TLC, column, HPLC, and LC-MS) and spectral studies such as FT-IR, CHNS analysis, 1H-NMR, and 13C-NMR. The antimicrobial potential of isolated compounds was assessed according to the standard methods. The isolated compounds were identified as arborine and skimmianine, which exhibited a significant antibacterial effect with the lowest MIC and MBC values against MDR S. aureus and in vitro kinetic and protein leakage assays supported the antimicrobial activity. Significant morphological changes such as uneven cell surfaces and morphology, cell shrinkage, and cell membrane damages were observed in the MDR S. aureus upon the treatment of arborine and skimmianine. The present investigation concludes that the isolated arborine and skimmianine compounds from G. pentaphylla harbor a strong antibacterial activity against MDR S. aureus and may be used as alternative natural drugs in the treatment of MDR S. aureus.

Antioxidant and Antiproliferative Potential of Bioactive Molecules Ursolic Acid and Thujone Isolated from Memecylon edule and Elaeagnus indica and Their Inhibitory Effect on Topoisomerase II by Molecular Docking Approach.

The present study aimed to evaluate the antioxidant and antiproliferative potential of ursolic acid and thujone isolated from leaves of Elaeagnus indica and Memecylon edule and their inhibitory effect on topoisomerase II using molecular docking study. The isolated ursolic acid and thujone were examined for different types of free radicals scavenging activity, the antiproliferative potential on U-937 and HT-60 cell lines by adopting standard methods. Further, these compounds were docked with the active site of the ATPase region of topoisomerase II. The findings of the research revealed that ursolic acid harbor strong antioxidant and antiproliferative capacity with low IC50 values than the thujone in all tested methods. Moreover, ursolic acid shows significant inhibition effect on topoisomerase II with a considerable docking score (-8.0312) and GLIDE energy (-51.86 kca/mol). The present outcome concludes that ursolic acid possesses significant antioxidant and antiproliferative potential, which can be used in the development of novel antioxidant and antiproliferative agents in the future.

Toxicological evaluation of lotus, ginkgo, and garlic tailored fermented Korean soybean paste (Doenjang) for biogenic amines, aflatoxins, and microbial hazards.

The present study aimed to develop a consortium of nutritive fermented food products, supplemented with phytochemicals, with reduced toxicological contents. We developed new flavored Doenjang products (protein rich) fermented with lotus, ginkgo, and garlic plant extract-based Meju (termed as EMD) as the starter culture and by using traditional Meju (termed as TMD), where these plant extracts were added later during the fermentation process. Fermented Doenjang samples were analyzed for reduced levels of biogenic amines (BAs), aflatoxins, and microbial hazards, (including Bacillus cereus) as well as for their nutritive contents and antioxidant potential, after varying periods of fermentation (0, 3, 6, 9 and 12 months). All Doenjang samples prepared using plant extracts and their mixtures (1% and 10%) showed desired reduction in B. cereus counts, BAs, aflatoxins, and other foodborne pathogens as well as showed potent antioxidant abilities, including phenolic/flavonoid contents. Based on the higher efficiency in reducing various toxicants, Ginkgo biloba leaf extract added TMD samples were selected for the development of Doenjang products as an innovative approach, with great potential to improve the quality and safety of soybean fermented products in the Korean market, offering enhanced health benefits and reduced risks of toxicity.

Garlic augments the functional and nutritional behavior of Doenjang, a traditional Korean fermented soybean paste.

Three different forms of garlic, namely, fresh garlic (2%, 6%, 10%), heat-dried (1%, 2%, 3%) and freeze-dried (1%, 2%, 3%), were supplemented in soybean paste to prepare Doenjang and further evaluated for functional, nutritional and safety aspects. Results showed a considerable antioxidant and anti-proliferative activity of garlic-supplemented Doenjang. As a measure of nutritive value, a high amount of total free amino acids, 4,290.73 mg/100 g-5,492.94 mg/100 g, was observed in prepared Doenjang. Among all preparations, 3% freeze-dried garlic-supplemented Doenjang proved the most effective against gastric adenocarcinoma and lung adenocarcinoma with 50% inhibition concentration of 7.66 ± 0.53 mg/mL and 7.82 ± 0.34 mg/mL, respectively. However 10% fresh-garlicsupplemented Doenjang (GGD-10) showed better activity against colorectal adenocarcinoma (HT29) cell line. Furthermore, GGD-10 effectively reduced colony formation and altered mitochondrial membrane potential of HT29 cells. Absence of pathogenic bacteria (Staphylococcus aureus, Salmonella species and Bacillus cereus) and aflatoxin was observed in Doenjang samples. In addition, nontoxic amount of anti-nutritional biogenic amines was observed in all the samples. The results collectively suggest that the addition of garlic in Doenjang can improve its nutritional and functional value, resulting in the protection of consumers from protein deficiencies and various stress conditions.

Detection of biogenic amines and microbial safety assessment of novel Meju fermented with addition of Nelumbo nucifera, Ginkgo biloba, and Allium sativum.

Meju, a cooked and fermented soy bean based food product, is used as a major ingredient in Korean traditional fermented foods such as Doenjang. We developed a novel type of Meju using single and combined extracts of Allium sativum (garlic clove), Nelumbo nucifera (lotus leaves), and Ginkgo biloba (ginkgo leaves) at 1% and 10% concentrations to improve the safety of Meju-based fermented products. Biogenic amines (BAs) in protein-rich fermented food products pose considerable toxical risks. The objective of this study was to investigate the effects of adding selected plant extracts in Meju samples during fermentation. Nine BAs, including tryptamine, 2-phenylethylamine, putrescine, cadaverine, agmatine, histamine, tyramine, spermidine and spermine, were isolated from Meju samples after sample derivatization with dansyl chloride and analyzed by high performance liquid chromatography. As a result, all tested Meju samples with added plant extracts showed total BAs levels in the range of 20.12 ±â€¯2.03 to 118.42 ±â€¯10.68 mg/100 g, which were below the safety limit set by various regulatory authorities (USFDA/KFDA/EFSA). However, among all tested Meju samples, LOM10 (Meju fermented with Nelumbo nucifera at 10% concentration) showed higher levels of BAs content than others either due to batch-to-batch variability or reduced beneficial microorganisms and/or due to increase in BA forming microorganisms. Also, none of the samples showed the aflatoxin level above the detection limit. Furthermore, all the tested Meju samples improved microbial safety as confirmed by the complete absence of Salmonella species and Staphylococcus aureus. However, some of the Meju samples showed the presence of coliforms (in range of 1.6 × 100-1.1 × 103 CFU/g), which is under regulatory limits. These results suggested that the use of plant extracts in Meju during fermentation have potential to improve microbial and toxicological safety of Meju products.

Development of novel Meju starter culture using plant extracts with reduced Bacillus cereus counts and enhanced functional properties.

We developed a novel type of Meju starter culture using single and combined extracts of Allium sativum (garlic clove), Nelumbo nucifera (lotus leaves), and Ginkgo biloba (ginkgo leaves) to improve the quality and functionality of Meju-based fermented products. Meju samples fermented with plant extracts (10 mg/ml) showed phenolic contents of 11.4-31.6 mg/g (gallic acid equivalents). Samples of extracts (garlic clove, lotus leaves, ginkgo leaves and their combination) fermented with Meju strongly inhibited tyrosinase, α-glucosidase, and elastase activities by 36.43-64.34%, 45.08-48.02%, and 4.52-10.90%, respectively. Specifically, ginkgo leaves extract added to fermented Meju samples at different concentrations (1% and 10%) strongly inhibited tyrosinase, α-glucosidase, and elastase activities and exhibited a potent antibacterial effect against Bacillus cereus with a significant reduction in bacterial counts compared with the effects observed for garlic clove and lotus leaf added to Meju samples. Scanning electron microscopy revealed severe morphological alterations of the B. cereus cell wall in response to ginkgo leaf extracts. Gas chromatographic mass spectroscopic analysis of plant extract-supplemented Meju samples and control Meju samples identified 113 bioactive compounds representing 98.44-99.98% total extract. The proposed approach may be useful for the development of various fermented functional foods at traditional and commercial levels.

Anti-angiogenic effect of Nelumbo nucifera leaf extracts in human umbilical vein endothelial cells with antioxidant potential.

Nelumbo nucifera Gaertn (Nymphaeaceae) has long been used as a traditional herb in Chinese, Japanese, Indian, and Korean medicinal practices since prehistoric times and flourishes today as the primary form of medicine. This study reports for the first time the potent ability of N. nucifera leaf extracts to inhibit vascular endothelial growth factor (VEGF)-induced angiogenesis in vitro and in vivo, as well as their antioxidant efficacy in various scavenging models and an analysis of their chemical composition. In vivo anti-angiogenic activity was evaluated in a chick chorioallantoic membrane (CAM) model using fertilized chicken eggs, in human umbilical vein endothelial cells (HUVECs) by using cell viability, cell proliferation and tube formation assays, and by determining intracellular reactive oxygen species (ROS) in vitro. The antioxidant efficacy of N. nucifera leaf extracts was determined in various scavenging models, including total phenolic and flavonoid content. The chemical composition of N. nucifera leaf extracts was determined by GC-MS analysis, which revealed the presence of different phytochemicals. The IC50 values for the DPPH radical scavenging activities of water and methanol extracts were found to be 1699.47 and 514.36 µg ml(-1), and their total phenolic and flavonoid contents were 85.01 ± 2.32 and 147.63 ± 2.23 mg GAE g dry mass(-1) and 35.38 ± 1.32 and 41.86 ± 1.07 mg QA g dry mass(-1), respectively. N. nucifera leaf extracts (10-100 µg ml(-1)) exhibited significant dose-dependent inhibition of VEGF-induced angiogenesis, as well as VEGF-induced proliferation and tube formation in HUVECs. In this study, N. nucifera leaf extracts displayed potent antioxidant and inhibitory effects on VEGF-induced angiogenesis. N. nucifera exerted an inhibitory effect on VEGF-induced proliferation and tube formation, as well as CAM angiogenesis in vivo. Moreover, N. nucifera leaf extracts significantly blocked VEGF-induced ROS production in HUVECs, confirming their possible anti-angiogenic mechanism.

Physiological and biochemical characterization of three nucleoside diphosphate kinase isozymes from rice (Oryza sativa L.).

Nucleoside diphosphate kinase (NDPK) is a ubiquitous enzyme that catalyzes the transfer of the γ-phosphoryl group from a nucleoside triphosphate to a nucleoside diphosphate. In this study, we examined the subcellular localization, tissue-specific gene expression, and enzymatic characteristics of three rice NDPK isozymes (OsNDPK1-OsNDPK3). Sequence comparison of the three OsNDPKs suggested differential subcellular localization. Transient expression of green fluorescence protein-fused proteins in onion cells indicated that OsNDPK2 and OsNDPK3 are localized to plastid and mitochondria respectively, while OsNDPK1 is localized to the cytosol. Expression analysis indicated that all the OsNDPKs are expressed in the leaf, leaf sheath, and immature seeds, except for OsNDPK1, in the leaf sheath. Recombinant OsNDPK2 and OsNDPK3 showed lower optimum pH and higher stability under acidic pH than OsNDPK1. In ATP formation, all the OsNDPKs displayed lower K(m) values for the second substrate, ADP, than for the first substrate, NTP, and showed lowest and highest K(m) values for GTP and CTP respectively.

Antimicrobial activities of 1,4-benzoquinones and wheat germ extract.

We evaluated the antibacterial activities of selected edible Korean plant seeds against the food-borne pathogens Staphylococcus aureus KCTC1927, Escherichia coli KCTC2593, Salmonella typhimurium KCTC2054, and Bacillus cereus KCTC1014. While screening for antibacterial agents, we discovered that wheat germ extract contains 2,6-dimethoxy-1,4-benzoquinone (DMBQ) and is highly inhibitory to S. aureus and B. cereus. This is the first report of the antibacterial activity of wheat germ extract. We also investigated the antibacterial activities of the 1,4- benzoquinone standards 1,4-benzoquinone (BQ), hydroquinone (HQ), methoxybenzoquinone (MBQ), and 2,6-dimethoxy- 1,4-benzoquinone (DMBQ). DMBQ and BQ were the most highly inhibitory to S. aureus and S. typhimurium, followed by MBQ and HQ. MICs for DMBQ and BQ ranged between 8 and 64 microgram/ml against the four foodborne pathogens tested. DMBQ and BQ showed significant antibacterial activity; the most sensitive organism was S. aureus with an MIC of 8 microgram/ml. BQ exhibited good activity against S. typhimurium (32 microgram/ml) and B. cereus (32 microgram/ml). The results suggest that wheat germ extract has potential for the development of natural antimicrobials and food preservatives for controlling foodborne pathogens.

Determination of non-volatile and volatile organic acids in Korean traditional fermented soybean paste (Doenjang).

Organic acids are formed in food as a result of metabolism of large molecular mass compounds. These organic acids play an important role in the taste and aroma of fermented food products. Doenjang is a traditional Korean fermented soybean paste product that provides a major source of protein. The quantitative data for volatile and non-volatile organic acid contents of 18 samples of Doenjang were determined by comparing the abundances of each peak by gas (GC) and high performance liquid chromatography (HPLC). The mean values of volatile organic acids (acetic acid, butyric acid, propionic acid and 3-methyl butanoic acid), determined in 18 Doenjang samples, were found to be 91.73, 29.54, 70.07 and 19.80 mg%, respectively, whereas the mean values of non-volatile organic acids, such as oxalic acid, citric acid, lactic acid and succinic acid, were noted to be 14.69, 5.56, 9.95 and 0.21 mg%, respectively. Malonic and glutaric acids were absent in all the tested samples of Doenjang. The findings of this study suggest that determination of organic acid contents by GC and HPLC can be considered as an affective approach to evaluate the quality characteristics of fermented food products.

The analysis of copper, selenium, and molybdenum contents in frequently consumed foods and an estimation of their daily intake in korean adults.

This study aimed to analyze the amounts of copper, selenium, and molybdenum among trace minerals in foods and to evaluate their daily intakes in Korean adults. Contents of copper, selenium, and molybdenum in 366 varieties of foods commonly consumed by Koreans were analyzed using inductively coupled plasma atomic emission spectrometry and inductively coupled plasma mass spectrometry, techniques with low detection limits as well as high reproducibility and precision. Next, we evaluated the status of trace mineral intake using the 24-h recall method after conducting anthropometric measurements on 249 male and 344 female adults aged 20 or older. The average daily energy intake for males was 7,452.8 kJ, significantly higher than the 6,118.3 kJ for females (p < 0.001). The average daily copper, selenium, and molybdenum intakes by males were 1,156.7, 135.5, and 12.2 microgg, respectively, compared to 1,028.5, 122.9, and 10.1 microg, respectively, by females. In males, the intake levels of copper and molybdenum were both significantly higher than in females. By continuously evaluating intake levels in this manner, it is anticipated that reference intakes of trace minerals will be established.