Search

Foam cell formation is a crucial process in the progression of atherosclerosis, a chronic inflammatory disease characterized by the accumulation of plaques in the arteries. This study aimed to examine the physiochemical changes in Laminaria japonica during fermentation with Bacillus subtilis and assess the inhibitory effects of the fermented sample on the oxidation of low-density lipoprotein (LDL) and the formation of foam cells. Polysaccharide decompositions and increase of total phenols and reducing power of L. japonica were observed during fermentation. The Fourier-transform infrared spectroscopy analysis revealed the peaks of seaweed polysaccharides of the fermented L. japonica (F sample). The presence of F sample reduced the oxidative stress induced by Cu2+ on LDL and inhibited foam cell formation. Additionally, the levels of proteins involved in foam cell activation, such as CD36, scavenger receptor-A, TLR4, and TLR6, were significantly decreased. Further proteomic analysis revealed that out of 1439 proteins up-regulated by oxidized LDL (oxLDL) in macrophages, 953 were downregulated in the presence of the F sample. The Ingenuity Pathway Analysis identified the activation of canonical pathways associated with atherosclerosis by oxLDL, which were subsequently suppressed by the F sample treatment. Protein-protein interaction network analysis uncovered the role of MAPK3 in regulating multiple pathways. This study suggests that B. subtilis-fermented brown seaweed L. japonica can inhibit LDL oxidation and foaming cell production. Additionally, it offers mechanistic insights for future research in this area.

ABSTRACTRecently, there have been reports on the immunomodulatory properties of polysaccharides (PP), oligosaccharides (PO), and mixtures of PO/PP derived from Porphyra. To explore their potential in alleviating allergic responses and enteric dysbiosis, this study delved deeper into their effects using a murine model of food allergy. Daily administration of Porphyra samples mitigated allergic diarrhea, enteritis, and allergen-specific IgE production. Moreover, Porphyra samples exerted a significant reduction in the secretion of IL-2, IL-4, IFN-γ, and IL-17A, along with an elevation in the IL-10/IL-4 ratio in splenocytes upon allergen stimulation. Similarly, expression of IL-4 and IFN-γ in the duodenum was suppressed. Notably, allergic mice displayed a distinct fecal microflora profile compared to that of normal mice. Interestingly, PP and PO treatments exhibited different effects on improving enteric dysbiosis. These findings collectively suggest the potential of PP and PO as functional food candidates for the management of allergic enteritis and dysbiosis.

The interactions among agar, gellan gum, gelatin, and modified waxy corn starch in the formation of mixed gels were examined in five different ratios. Binary hydrocolloid gels were prepared using three ingredients: two hydrocolloids (total hydrocolloid concentration: 0.5 wt%, ratios of mixture: 0/0.5, 0.1/0.4, 0.2/0.3, 0.3/0.2, 0.4/0.1, and 0.5/0) and water. The textural properties of the hydrocolloid gels were studied by measuring the gel strength, rigidity, breaking force, breaking point, and syneresis as functions of the mixing ratio. The higher syneresis percentage of binary modified waxy corn starch and gum gels than that of mixed gum gels after cold storage was mainly due to the retrogradation of amylopectin. Agar was shown be the most influential with regards to increasing the gel strength, breaking force, and rigidity among the three kinds of gum, while gellan gum was more effective against syneresis than agar and gelatin for storage periods of 7 and 14 days. In the mixed gels, a dramatic increase in the breaking point from 0 to 0.5% was only exhibited for gellan gum. The results provided useful information, including gel strength, rigidity, breaking force, breaking point, and syneresis, for gum and modified corn starch ingredients selected from refrigerated binary gum gels such as pudding for food product development.

This study examined the physicochemical effects of the fortification of noodles with 0.25–1.00% (w/w) calcium salts, viz. calcium acetate, calcium carbonate, calcium citrate, and calcium lactate. Fortification with calcium citrate, calcium acetate, and calcium carbonate increased the pH and breaking force of the dried noodles. However, the fortification of noodles with any concentration of calcium did not increase the extent of elongation of the control raw noodles. The L* and b* values of the raw and dried noodle color increased with increasing concentrations of calcium salts, except for noodles with added calcium citrate. Fortification with calcium citrate yielded no significant influence on color, texture, adhesiveness, springiness, flavor, and overall scores for cooked noodles. Noodles fortified with 0.5% calcium citrate made from oyster shells were compared with a control sample of noodles and noodles fortified with commercially available calcium citrate. The particle size of the calcium citrate made from oyster shells (258 nm) was smaller than that of the purchased calcium citrate (2631 nm). Noodles fortified with calcium citrate made from oyster shells showed no significantly difference compared to noodles fortified with commercially available calcium citrate. These results suggest that calcium citrate made from oyster shells may be used as the additive of choice for the manufacture of calcium-fortified noodles.

This study aims to evaluate the effects of different softening methods on the texture and quality of abalones (Haliotis discus hannai × Diversicolor diversicolor). We monitored the hardness, meat weight, pH, size, and appearance from the sterilized abalones pre-treated with alkaline soaking and enzymatic injection/soaking, with either vacuum orbital shaking or ultrasonic processing. The hardness of the abalones was reduced by alkaline and enzymatic treatments, and combined vacuum orbital shaking or ultrasound treatment and this could increase their tendering effects. The softest abalone was obtained by bromelain comparing to papain injection, and the TVBN measurements indicated that the abalones remained in a fresh state under all treatments. Both enzymatic treatments showed a dose-dependent effects on hardness and pH of sterilized abalones. The abalones pre-treated with injecting enzyme exhibited significant softening effect than soaking method. Histological observation indicated that severe fracturing and non-uniform large spaces between muscle fibers were observed in the abalones injected with bromelain, and the abalones subjected to NaHCO3 soaking showed distorted muscle fibers and small uniform spaces. SDS-PAGE further indicated the decrease of paramyosin in the abalones injected with bromelain. Sensory evaluation using senior panelists without chewing difficulties indicated that NaHCO3 soaking before sterilization exhibited the highest overall acceptability, possibly because this method could slightly soften abalone foot while retaining its palatability. This study should benefit the food industry in terms of developing and processing food for senior citizens.

Seaweeds have a variety of biological activities, and their aromatic characteristics could play an important role in consumer acceptance. Here, changes in aroma compounds were monitored during microbial fermentation, and those most likely to affect sensory perception were identified. Ulva sp. and Laminaria sp. were fermented and generally recognized as safe microorganisms, and the profile of volatile compounds in the fermented seaweeds was investigated using headspace solid-phase microextraction with gas chromatography–mass spectrometry. Volatile compounds, including ketones, aldehydes, alcohols, and acids, were identified during seaweed fermentation. Compared with lactic acid bacteria fermentation, Bacillus subtilis fermentation could enhance the total ketone amount in seaweeds. Saccharomyces cerevisiae fermentation could also enhance the alcohol content in seaweeds. Principal component analysis of volatile compounds revealed that fermenting seaweeds with B. subtilis or S. cerevisiae could reduce aldehyde contents and boost ketone and alcohol contents, respectively, as expected. The odor of the fermented seaweeds was described by using GC–olfactometry, and B. subtilis and S. cerevisiae fermentations could enhance pleasant odors and reduce unpleasant odors. These results can support the capability of fermentation to improve the aromatic profile of seaweeds.

The World Health Organization indicated that antibiotic resistance is one of the greatest threats to health, food security, and development in the world. Drug resistance efflux pumps are essential for antibiotic resistance in bacteria. Here, we evaluated the plant phenolic compound ethyl 3,4-dihydroxybenzoate (EDHB) for its efflux pump inhibitory (EPI) activity against drug-resistant Escherichia coli. The half-maximal inhibitory concentration, modulation assays, and time-kill studies indicated that EDHB has limited antibacterial activity but can potentiate the activity of antibiotics for drug-resistant E. coli. Dye accumulation/efflux and MALDI-TOF studies showed that EDHB not only significantly increases dye accumulation and reduces dye efflux but also increases the extracellular amount of antibiotics in the drug-resistant E. coli, indicating its interference with substrate translocation via a bacterial efflux pump. Molecular docking analysis using AutoDock Vina indicated that EDHB putatively posed within the distal binding pocket of AcrB and in close interaction with the residues by H-bonds and hydrophobic contacts. Additionally, EDHB showed an elevated postantibiotic effect on drug-resistant E. coli. Our toxicity assays showed that EDHB did not change the bacterial membrane permeability and exhibited mild human cell toxicity. In summary, these findings indicate that EDHB could serve as a potential EPI for drug-resistant E. coli.

The objectives of this study were to evaluate the effect of 0.5% chitosan incorporation on acrylamide development in a food model solution containing 0.5% fructose and asparagine after heating for 30 min at 180 °C. All the solutions were investigated for the following characteristics: acrylamide, asparagine, reducing sugar content, color, kinematic viscosity, Maillard reaction products (MRPs), and pH every 10 min. After heating for 10 min, the viscosity of chitosan-containing solutions reduced significantly. The investigational data confirmed that chitosan may have decomposed into lower molecular structures, as demonstrated by the reduced viscosity of the solution at pH < 6 and a decrease in the acrylamide content during 30 min of heating in a fructose–asparagine system. This study also confirms that the formation of ultraviolet-absorbing intermediates and browning intensity of MRPs containing acrylamide prepared by fructose–asparagine was more than those of MRPs prepared by glucose–asparagine solution system. MRPs containing acrylamide resulted from the reaction of asparagine with fructose (ketose) rather than glucose (aldose). Acrylamide formation could be significantly mitigated in the fructose–asparagine–chitosan model system as compared to the fructose–asparagine model system for possible beverage and food application.

We have investigated different properties (thickness, moisture loss, oil uptake, breaking force, color, puffing ratio during 0.5–5 min frying, microstructure, and sensory evaluation) of raw pork skins with varying thickness (2, 3, and 4 mm) after drying, intended as deep-fried snacks. We have found that the oil content, breaking force, and puffing ratio of fried pork skin with different raw skin thickness have no significant difference under similar water content (1.68–1.98 g/100 g wet weight basis, wb) after 3–5 min of deep-frying at 180 °C. Additionally, sensory score results have shown that fried pork skins with 4 mm raw skin thickness had lower flavor, texture, and overall acceptability than those with 2 mm and 3 mm raw skin thickness. Scanning electron micrographs (SEM) have revealed less holes and irregular and crack microstructure in fried pork skins with 4 mm raw skin thickness than in other groups. Different thickness of raw pork skins resulted in different effects in microstructure and influenced water evaporation and oil uptake of fried pork skin. Finally, we have proposed the kinetic equations of water loss and oil uptake of fried pork skins. Fried pork skin from raw skin thicker than 4 mm need frying at temperature higher than 180 °C to improve their puffing ratio and sensory acceptability.

The fat content of fried pork rinds is high, and alternative frying helps reduce the oil content and maintain their texture and taste. Different frying methods such as microwave-assisted, traditional deep frying and vacuum frying on the breaking force, color, microstructure, water loss and oil absorption attributes of fried pork rinds were evaluated in this study. The fat content of microwave-assisted and vacuum-fried pork rinds was lower (24.2 g/100 g dry weight basis (db) and 17.1 g/100 g db, respectively) than that (35.6 g/100 g db) of traditional deep-fat frying. Non-uniform, holy and irregular surface microstructures were obtained by vacuum frying due to rapid mass transfer at low pressure. The first-order kinetic models of water loss and oil absorption of traditional and microwave-assisted frying of pork rinds were established. Microwave frying caused a faster moisture loss rate, shorter frying time and lower pork rind oil content, makes it an attractive substitute for traditional deep-fat frying.

Drug efflux pumps are one of the major elements used by antibiotic-resistant bacteria. Efflux pump inhibitors (EPIs) are potential therapeutic agents for adjunctive therapy, which can restore the activity of antibiotics that are no longer effective against pathogens. This study evaluated the seaweed compound diphenylmethane (DPM) for its EPI activity. The IC50 and modulation results showed that DPM has no antibacterial activity but can potentiate the activity of antibiotics against drug-resistant E. coli. Time-kill studies reported that a combination of DPM and erythromycin exhibited greater inhibitory activity against drug-resistant Escherichia coli. Dye accumulation and dye efflux studies using Hoechst 33342 and ethidium bromide showed that the addition of DPM significantly increased dye accumulation and reduced dye efflux in drug-resistant E. coli, suggesting its interference with dye translocation by an efflux pump. Using MALDI-TOF, it was observed that the addition of DPM could continuously reduce antibiotic efflux in drug-resistant E. coli. Additionally, DPM did not seem to damage the E. coli membranes, and the cell toxicity test showed that it features mild human-cell toxicity. In conclusion, these findings showed that DPM could serve as a potential EPI for drug-resistant E. coli.

Seaweed oligosaccharides have attracted attention in food, agricultural, and medical applications recently. Compared to red and brown seaweeds, fewer studies have focused on the biological activity of green seaweed’s oligosaccharides. This study aimed to produce bioactive ulvan oligosaccharides via enzymatic hydrolysis from green seaweed Ulva lactuca. Ulvan, a water-soluble polysaccharide, was obtained by hot water extraction. Two isolated marine bacteria, Pseudomonas vesicularis MA103 and Aeromonas salmonicida MAEF108, were used to produce multiple hydrolases, such as ulvanolytic enzymes, amylase, cellulase, and xylanase, to degrade the ulvan extract. An ultrafiltration system was used to separate the enzymatic hydrolysate to acquire the ulvan oligosaccharides (UOS). The characteristics of the ulvan extract and the UOS were determined by yield, reducing sugar, uronic acid, sulfate group, and total phenols. The FT-IR spectrum indicated that the ulvan extract and the UOS presented the bands associated with O-H, C=O, C-O, and S=O stretching. Angiotensin I converting enzyme (ACE) inhibition and antioxidant activities in vitro were evaluated in the ulvan extract and the UOS. These results provide a practical approach to producing bioactive UOS by microbial enzymatic hydrolysis that can benefit the development of seaweed-based products at the industrial scale.

4-Mercapto-4-methyl-2-pentanone (4MMP), a high-impact aroma compound with the box tree and black currant flavors was first identified in wines and could be released by microbial cysteine-S-conjugate β-lyases from its precursors. In this study, various yeasts and bacteria encoding β-lyases were selected to examine their β-lyase activities. A thiol precursor of 4MMP, cysteine-conjugate of 4MMP (cys-4MMP), was synthesized with a purity of >95% in a relatively environmentally friendly approach, and its chemical structure was confirmed by nuclear magnetic resonance spectroscopy. The β-lyase activities of the crude cell extract from the bacteria and yeast strains for different substrates were examined using a colorimetric method. Shewanella putrefaciens cell extract exhibited the highest β-lyase activity for all tested substrates. Additionally, the optimum pH and temperature for their β-lyase activities were determined. To monitor the conversion efficiency of precursor cys-4MMP to 4MMP, liquid chromatography-mass spectrometry was used. Our data indicate that selected bacteria and yeasts could convert cys-4MMP into 4MMP, and S. putrefaciens exhibited the best conversion yield. This study demonstrated the potential use of microbial cell extracts to produce sulfur-containing aroma compounds such as 4MMP.

The aim of this research was to investigate the effects of the addition of 0.5% hydroxymethylfurfural (HMF) and low molecular chitosan on acrylamide and HMF formation in a food model system, which contains 0.5% glucose, asparagine, and HMF within 30 min of heating at 180 °C. At an interval of 10 min, all solutions were evaluated in the following aspects: reducing sugar, asparagine, acrylamide, HMF content, pH, Maillard reaction products, kinematic viscosity, and color. After heating for 10 min, the kinematic viscosity of solutions containing chitosan reduced significantly. The values of the acrylamide, HMF, and absorbance increased at OD294 and OD420 (optical density measured at 294 nm and 420 nm) of solutions. Experimental results showed that low-molecular-weight chitosan might be hydrolyzed into much lower molecular weight, followed by the decrease in kinematic viscosity of the solution at pH lower than 6 and the increase in the formation of acrylamide after heating for 30 min.

This research focuses on the proteolytic capacity of sea bass byproduct (SB) and their hypocholesterolemic activity via the cholesterol micelle formation (CMF) inhibition. SB was fermented with seven mixed lactic acid bacteria for 5 h at 42 °C. The lactic fermented SB was hydrolyzed with Protease N for 6 h under HHP to obtain the SB hydrolysates (HHP-assisted Protease N hydrolysis after fermentation, F-HHP-PN6). The supernatant was separated from the SB hydrolysate and freeze-dried. As the hydrolysis time extended to 6 h, soluble protein content increased from 187.1 to 565.8 mg/g, and peptide content increased from 112.8 to 421.9 mg/g, while inhibition of CMF increased from 75.0% to 88.4%. Decreasing the CMF inhibitory activity from 88.4% to 42.1% by simulated gastrointestinal digestion (FHHP-PN6 was further hydrolyzed by gastrointestinal enzymes, F-HHP-PN6-PP) reduced the CMF inhibitory activity of F-HHP-PN6. Using gel filtration chromatography, the F-HHP-PN6-PP was fractioned into six fractions. The molecular weight of the fifth fraction from F-HHP-PN6-PP was between 340 and 290 Da, and the highest inhibitory efficiency ratio (IER) on CMF was 238.9%/mg/mL. Further purification and identification of new peptides with CMF inhibitory activity presented the peptide sequences in Ser-Ala-Gln, Pro-Trp, and Val-Gly-Gly-Thr; the IERs were 361.7, 3230.0, and 302.9%/mg/mL, respectively.

Multidrug efflux pumps play an essential role in antibiotic resistance. The conventional methods, including minimum inhibitory concentration and fluorescent assays, to monitor transporter efflux activity might have some drawbacks, such as indirect evidence or interference from color molecules. In this study, MALDI-TOF MS use was explored for monitoring drug efflux by a multidrug transporter, and the results were compared for validation with the data from conventional methods. Minimum inhibitory concentration was used first to evaluate the activity of Escherichia coli drug transporter AcrB, and this analysis showed that the E. coli overexpressing AcrB exhibited elevated resistance to various antibiotics and dyes. Fluorescence-based studies indicated that AcrB in E. coli could decrease the accumulation of intracellular dyes and display various efflux rate constants for different dyes, suggesting AcrB’s efflux activity. The MALDI-TOF MS analysis parameters were optimized to maintain a detection accuracy for AcrB’s substrates; furthermore, the MS data showed that E. coli overexpressing AcrB led to increased ions abundancy of various dyes and drugs in the extracellular space at different rates over time, illustrating continuous substrate efflux by AcrB. This study concluded that MALDI-TOF MS is a reliable method that can rapidly determine the drug pump efflux activity for various substrates.

Biodegradable polylactic acid material is manufactured from lactic acid, mainly produced by microbial fermentation. The high production cost of lactic acid still remains the major limitation for its application, indicating that the cost of carbon sources for the production of lactic acid has to be minimized. In addition, a lack of source availability of food crop and lignocellulosic biomass has encouraged researchers and industries to explore new feedstocks for microbial lactic acid fermentation. Seaweeds have attracted considerable attention as a carbon source for microbial fermentation owing to their non-terrestrial origin, fast growth, and photoautotrophic nature. The proximate compositions study of red, brown, and green seaweeds indicated that Gracilaria sp. has the highest carbohydrate content. The conditions were optimized for the saccharification of the seaweeds, and the results indicated that Gracilaria sp. yielded the highest reducing sugar content. Optimal lactic acid fermentation parameters, such as cell inoculum, agitation, and temperature, were determined to be 6% (v/v), 0 rpm, and 30 °C, respectively. Gracilaria sp. hydrolysates fermented by lactic acid bacteria at optimal conditions yielded a final lactic acid concentration of 19.32 g/L.

Nervous necrosis virus (NNV) results in high mortality rates of infected marine fish worldwide. Interferons (IFNs) are cytokines in vertebrates that suppress viral replication and regulate immune responses. Heterologous overexpression of fish IFN in bacteria could be problematic because of protein solubility and loss of function due to protein misfolding. In this study, a protein model of the IFN-α of Epinephelus septemfasciatus was built based on comparative modeling. In addition, PelB and SacB signal peptides were fused to the N-terminus of E. septemfasciatus IFN-α for overexpression of soluble, secreted IFN in Escherichia coli (E-IFN) and Bacillus subtilis (B-IFN). Cytotoxicity tests indicated that neither recombinant grouper IFN-α were cytotoxic to a grouper head kidney cell line (GK). The GK cells stimulated with E-IFN and B-IFN exhibited elevated expression of antiviral Mx genes when compared with the control group. The NNV challenge experiments demonstrated that GK cells pretreated or co-treated with E-IFN and B-IFN individually had three times the cell survival rates of untreated cells, indicating the cytoprotective ability of our recombinant IFNs. These data provide a protocol for the production of soluble, secreted, and functional grouper IFN of high purity, which may be applied to aquaculture fisheries for antiviral infection.

The gene agaA, of the isolated marine bacterium Pseudomonas vesicularis MA103, comprised 2958-bp nucleotides encoding a putative agarase AgaA of 985 amino acids, which was predicted to contain a signal peptide of 29 amino acids in the N-terminus, a catalytic domain of glycoside hydrolase 16 (GH16) family, a bacterial immunoglobulin group 2 (Big 2), and three carbohydrate binding modules 6 (CBM 6). The gene agaA was cloned and overexpressed in Escherichia coli, and the optimum temperatures for AgaA overexpression were 16, 20 and 24 °C. The agaA was cloned without its signal peptide for cytosolic production overexpression, whereas it was cloned with the heterologous signal peptide PelB and its endogenous signal peptide for periplasmic and extracellular productions, respectively. Extracellular and periplasmic rAgaA showed greater activity than that of cytosolic rAgaA, indicating that membrane translocation of AgaA may encourage proper protein folding. Time-course hydrolysis of agarose by rAgaA was accomplished and the products were analyzed using thin layer chromatography and matrix-assisted laser desorption inoization-time of flight mass spectrometry, indicating that AgaA from P. vesicularis was an endo-type β-1,4 agarase that cleaved agarose into neoagarotetraose and neoagarohexaose as the final products.

The genome sequence of V. cholerae O1 Biovar Eltor strain N16961 has revealed a putative antibiotic resistance (var) regulon that is predicted to encode a transcriptional activator (VarR), which is divergently transcribed relative to the putative resistance genes for both a metallo-β-lactamase (VarG) and an antibiotic efflux-pump (VarABCDEF). We sought to test whether these genes could confer antibiotic resistance and are organised as a regulon under the control of VarR. VarG was overexpressed and purified and shown to have β-lactamase activity against penicillins, cephalosporins and carbapenems, having the highest activity against meropenem. The expression of VarABCDEF in the Escherichia coli (ΔacrAB) strain KAM3 conferred resistance to a range of drugs, but most significant resistance was to the macrolide spiramycin. A gel-shift analysis was used to determine if VarR bound to the promoter regions of the resistance genes. Consistent with the regulation of these resistance genes, VarR binds to three distinct intergenic regions, varRG, varGA and varBC located upstream and adjacent to varG, varA and varC, respectively. VarR can act as a repressor at the varRG promoter region; whilst this repression was relieved upon addition of β-lactams, these did not dissociate the VarR/varRG-DNA complex, indicating that the de-repression of varR by β-lactams is indirect. Considering that the genomic arrangement of VarR-VarG is strikingly similar to that of AmpR-AmpC system, it is possible that V. cholerae has evolved a system for resistance to the newer β-lactams that would prove more beneficial to the bacterium in light of current selective pressures.

In this study, the low-molecular-weight (LMW) fucoidan, rich in fucose and sulfate, was extracted and purified from the edible brown seaweed, Laminaria japonica. In this study, we orally administered LMW fucoidan to mice for 6 weeks. We then examined fucoidan’s effects on innate immunity, adaptive immunity, and Mycoplasma pneumoniae (MP)-antigen-stimulated immune responses. Our data showed that LMW fucoidan stimulated the innate immune system by increasing splenocyte proliferation, natural killer (NK) cell activity, and phagocytic activity. LMW fucoidan also increased interleukin (IL)-2, IL-4, and interferon (IFN)-γ secretion by splenocytes and immunoglobulin (Ig)-G and IgA content in serum, which help regulate adaptive immune cell functions, and decreased allergen-specific IgE. In MP-antigen-stimulated immune responses, the IgM and IgG content in the serum were significantly higher in the LMW fucoidan group after MP-antigen stimulation. Our study provides further information about the immunomodulatory effects of LMW fucoidan and highlights a potential role in preventing M. pneumoniae infection.

Vibrio cholerae ATP-binding cassette transporter VcaM (V. cholerae ABC multidrug resistance pump) has previously been shown to confer resistance to a variety of medically important drugs. In this study, we set to analyse its properties both in vitro in detergent-solubilised state and in vivo to differentiate its dependency on auxiliary proteins for its function. We report the first detailed kinetic parameters of purified VcaM and the rate of phosphate (Pi) production. To determine the possible functional dependencies of VcaM on the tripartite efflux pumps we then utilized different E. coli strains lacking the principal secondary transporter AcrB (Acriflavine resistance protein), as well as cells lacking the outer membrane factor (OMF) TolC (Tolerance to colicins). Consistent with the ATPase function of VcaM we found it to be susceptible to sodium orthovanadate (NaOV), however, we also found a clear dependency of VcaM function on TolC. Inhibitors targeting secondary active transporters had no effects on either VcaM-conferred resistance or Hoechst 33342 accumulation, suggesting that VcaM might be capable of engaging with the TolC-channel without periplasmic mediation by additional transporters. Our findings are indicative of VcaM being capable of a one-step substrate translocation from cytosol to extracellular space utilising the TolC-channel, making it the only multidrug ABC-transporter outside of the MacB-family with demonstrable TolC-dependency.

The combined effects of low-molecular-weight fucoidan (LMF) and fucoxanthin (Fx) in terms of antihyperglycemic, antihyperlipidemic, and hepatoprotective activities were investigated in a mouse model of type II diabetes. The intake of LMF, Fx, and LMF + Fx lowered the blood sugar and fasting blood sugar levels, and increased serum adiponectin levels. The significant decrease in urinary sugar was only observed in LMF + Fx supplementation. LMF and Fx had ameliorating effects on the hepatic tissue of db/db mice by increasing hepatic glycogen and antioxidative enzymes, and LMF was more effective than Fx at improving hepatic glucose metabolism. As for glucose and lipid metabolism in the adipose tissue, the expression of insulin receptor substrate (IRS)-1, glucose transporter (GLUT), peroxisome proliferator-activated receptor gamma (PPARγ), and uncoupling protein (UCP)-1 mRNAs in the adipose tissue of diabetic mice was significantly upregulated by Fx and LMF + Fx, and levels of inflammatory adipocytokines, such as adiponectin, tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6), were significantly modulated only by LMF + Fx supplementation. The efficacy of LMF + Fx supplementation on the decrease in urinary sugar and on glucose and lipid metabolism in the white adipose tissue of db/db mice was better than that of Fx or LMF alone, indicating the occurrence of a synergistic effect of LMF and Fx.

For a long time, fucoidan has been well known for its pharmacological activities, and recently low molecular weight fucoidan (LMF) has been used in food supplements and pharmaceutical products. In the present study, LMF was extracted from Laminaria japonica by enzyme hydrolysis. The toxicity of LMF in mouse and rat models was determined by many methods, such as total arsenic content, bacterial reverse mutation assay, chromosome aberration assay, and in vivo micronucleus assay. The present findings showed that LMF at 5000 μg/mL exhibited no mutagenicity. It also produced no formatting disruption of red blood cells in vivo. At 2000 mg/kg BW/day there were no toxicological indications. LMF is expected to be used as a safe food supplement.