Agar-tragacanth/silk fibroin hydrogel containing Zn-based MOF as a novel nanobiocomposite with biological activity.

In this study, a novel nanobiocomposite consisting of agar (Ag), tragacanth gum (TG), silk fibroin (SF), and MOF-5 was synthesized and extensively investigated by various analytical techniques and basic biological assays for potential biomedical applications. The performed Trypan blue dye exclusion assay indicated that the proliferation percentage of HEK293T cells was 71.19%, while the proliferation of cancer cells (K-562 and MCF-7) was significantly lower, at 10.74% and 3.33%. Furthermore, the Ag-TG hydrogel/SF/MOF-5 nanobiocomposite exhibited significant antimicrobial activity against both E. coli and S. aureus strains, with growth inhibition rates of 76.08% and 69.19% respectively. Additionally, the hemolytic index of fabricated nanobiocomposite was found approximately 19%. These findings suggest that the nanobiocomposite exhibits significant potential for application in cancer therapy and wound healing.

Gelatin/agar pH-indicator film based on cranberry extract loaded with linalool nanoparticle: Survey on physical, antimicrobial, and antioxidant properties.

In this study, linalool-nanoparticles (L-NPs) were prepared (encapsulation efficiency was 68.54 %) and introduced pH-indicator film based on cranberry-extract (CEF) to develop multifunctional smart films. XRD analysis and FTIR spectroscopy indicated that cranberry-extract (CE) and L-NPs were uniformly distributed in the gelatin/agar matrix and could change the intermolecular structure of the film. Color change of smart films showed that CE endowed the film with pH-sensitive property. As CE and L-NPs were added to the film, the water contact angle (WCA) was increased from 57.03° to 117.73°, the elongation at break (EAB) was increased from 12.30 % to 34.60 %. Additionally, the introduction of L-NPs enhanced the antioxidant activity (DPPH free radical scavenging rate increased from 26.80 % to 36.35 %) and antibacterial activity (against S. aureus and E. coli) of the smart film, which were verified by its retarding effect on pork spoilage.

Real-time monitoring of pork freshness using polyvinyl alcohol/modified agar multilayer gas-sensitive labels.

Accurate consumer perception of food packages should provide real-time feedback on any changes inside food packaging. Hence, a new multilayer gas-sensitive label (POA-12) was prepared using a layer-by-layer pouring method for simple, visual, and real-time detection of pork's freshness, while the front side was developed by immobilizing red carbon dots and fluorescein isothiocyanate in POA as indicator for volatile nitrogen, and the back side was created using bromothymol blue in POA as pH indicator. The swelling index of the multilayer gas-sensitive labels reduced from 159.19% to 148.36%, and the tensile strength increased from 25.52 MPa to 42.61 MPa. In addition, the POA-12 multilayer label showed a red-to-yellow fluorescence change as TVB-N increased from 6.84 to 31.4 and a yellow-brown-to-blue-green color change as pH increased from 5.74 to 7.24 when detecting pork samples. Thus, it provides dual-indicator monitoring that improves the accuracy and reliability of assessing the freshness of high-protein products.

Headspace control and antimicrobials: Inhibition strategies to prevent the growth of Alicyclobacillus acidoterrestris in orange juice.

Alicyclobacillus acidoterrestris can cause spoilage in orange juice that leads to consumer rejection. Six different orange juices were physiochemically characterized (pH, total soluble solids, titratable acidity, total polyphenols and vitamin C). A bottle for each sampling point per juice was filled (headspace: 40% volume) and inoculated with 102 -103 CFU per ml of A. acidoterrestris ATCC® 49025™ (heat shocked before inoculation: 75°C, 20 min). Samples were stored for 21 days at 45 ± 1°C and plate counted periodically on acidified YSG agar (pH 3·7) incubated at 45 ± 1°C for 3 days. The effect of headspace (6% versus 40% volume) on A. acidoterrestris growth was also evaluated. The effect of nisin (0·006, 0·003, 0·0015, and 0·00075%), sodium benzoate (0·1%), potassium sorbate (0·1%) and a mix of benzoate and sorbate (0·05% each) on A. acidoterrestris was additionally addressed. Alicyclobacillus acidoterrestris reached up to 107 CFU per ml in five of the six juices in less than 1 week. Headspace significantly impacted (P < 0·05) A. acidoterrestris maximum population, which reached the critical value of 5 log CFU per ml at 40% headspace. All preservatives, regardless of concentration, showed a bacteriostatic effect during 22 days of storage with no significant differences amongst treatments (P > 0·05).

Emulsion gels and oil-filled aerogels as curcumin carriers: Nanostructural characterization of gastrointestinal digestion products.

Agar and κ-carrageenan emulsion gels and oil-filled aerogels were investigated as curcumin carriers and their structure and mechanical properties, as well as their structural changes upon in vitro gastrointestinal digestion were characterized. Agar emulsion gels presented stiffer behaviour, with smaller and more homogeneous oil droplets (ϕ âˆ¼ 12 µm) than those from κ-carrageenan (ϕ âˆ¼ 243 µm). The structure of κ-carrageenan gels was characterized by the presence of rigid swollen linear chains, while agar produced more branched networks. After simulated gastrointestinal digestion bile salt lamellae/micelles (∼5 nm) and larger vesicles of partially digested oil (Rg âˆ¼ 20-50 nm) were the predominant structures, being their proportion dependent of the polysaccharide type and the physical state of the gel network. The presence of curcumin induced the formation of larger vesicles and limited the formation of mixed lamellae/micelles.

Carrageenan/agar-based functional film integrated with zinc sulfide nanoparticles and Pickering emulsion of tea tree essential oil for active packaging applications.

A functional carrageenan/agar-based film was prepared by combining tea tree oil Pickering emulsion (PET) and zinc sulfide nanoparticles (ZnSNP). PET was formulated using tea tree essential oil stabilized with nanocellulose fibers. PET and ZnSNPs were uniformly dispersed in the binary polymer matrix and formed compatible films. The incorporation of ZnSNPs improved the mechanical strength, whereas PET slightly decreased the strength, but the combined addition of ZnSNP and PET maintained the mechanical strength with slightly improved flexibility. The addition of ZnSNP and PET, alone or in combination, slightly improved the water vapor barrier, water resistance, and thermal stability of the film. In addition, the carrageenan/agar-based composite membrane showed distinct antioxidant and antibacterial activity. The ZnSNP and PET incorporated binary composite films with enhanced physical and functional properties are likely to be used in active food packaging applications.

Simple stain-free screening method for pectinolytic microorganisms under alkalophilic conditions.

OBJECTIVES: To develop a simple pectin-degrading microorganism screening method. RESULTS: We developed a method utilizing the phenomenon whereby cooling an alkaline agar medium containing pectin causes the agar to become cloudy. This highly simplified method involves culturing the microorganisms on pectin-containing agar medium until colony formation is observed, and subsequent overnight cooling of the agar medium to 4 °C. Using this simple procedure, we successfully identified pectin-degrading microorganisms by observing colonies with halos on the clouded agar medium. We used alkaline pectinase and Bacillus halodurans, which is known to secrete alkaline pectinase, to establish the screening method. We demonstrated the screening of pectin-degrading microorganisms using the developed method and successfully isolated pectin-degrading microorganisms (Paenibacillus sp., Bacillus clausii, and Bacillus halodurans) from a soil sample. CONCLUSIONS: The developed method is useful for identifying pectin-degrading microorganisms.

A novel strategy of nanosized herbal Plectranthus amboinicus, Phyllanthus niruri and Euphorbia hirta treated TiO2 nanoparticles for antibacterial and anticancer activities.

Titanium dioxide nanoparticles exhibit good anticancer and antibacterial activities. They are known to be environmentally friendly, stable, less toxic, and have excellent biocompatibility nature. Due to these properties, they are well suited for biological applications particularly in biomedical applications such as drug delivery and cancer therapy. In this research article, three medicinal herbs namely, Plectranthus amboinicus (Karpooravalli), Phyllanthus niruri (Keezhanelli), and Euphorbia hirta (Amman Pacharisi), were used to modify the surface of the TiO2 nanoparticles. The synthesized nanoparticles were subjected to various characterization techniques. The samples are then subjected to MTT assay to determine cell viability. KB oral cancer cells are used for the determination of the anticancer nature of the pure and bio modified nanoparticles. It is observed that Plectranthus amboinicus-Phyllanthus niruri modified TiO2 nanoparticles exhibit excellent anticancer activities among other bio modified and pure samples. The samples are then examined for antibacterial activities against three Gram-negative bacterial strains namely, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and two Gram-positive bacterial strains namely, Staphylococcus aureus and Streptococcus mutans, respectively. Among the modified and pure samples, Plectranthus amboinicus showed good antibacterial activity against Gram-positive and Gram-negative bacteria. In the Flow cytometry analysis, the generation of p53 protein expression from Plectranthus amboinicus-Phyllanthus niruri modified TiO2 nano herbal particles shows the anti-cancerous nature of the sample. Then to determine the toxic nature of the Plectranthus amboinicus-Phyllanthus niruri modified TiO2 nano herbal particles against normal cells, the NPs were subjected to MTT assay against normal L929 cells, and it was found to be safer and less toxic towards the normal cells.

Dopamine/zinc oxide doped poly(N-hydroxyethyl acrylamide)/agar dual network hydrogel with super self-healing, antibacterial and tissue adhesion functions designed for transdermal patch.

Dual network (DN) hydrogels with excellent mechanical strength and controllable component adjustment characteristics have a broad application range in the field of biomedicine. However, the tissue adhesion, skin affinity, self-healing, and antibacterial properties of DN hydrogels are inadequate for their application as skin patches. In this work, we prepared dopamine/zinc oxide (DOPA/ZnO) doped poly(N-hydroxyethyl acrylamide) (p(HEAA))/agar DN hydrogels and combined them to obtain a bilayer hydrogel (two-layer gel) with moisturizing properties. Upon incorporating 0.86 wt% of dopamine (DOPA), the resultant DOPA/p(HEAA))/agar DN hydrogel (DOPA@DNG) exhibited high tensile strain (up to 1600%), excellent self-repair ability, and tissue adhesion. ZnO/p(HEAA))/agar DN hydrogel (ZnO NG) obtained by incorporating 2 w/v ZnO nanoparticles (ZnO NPs) achieved high tensile strength (1.2 MPa), good antibacterial ability, and low charge transfer resistance. Moreover, ZnO NG, which has a tight structure, was employed as a protective layer for the two-layer gel, which can effectively slow down the excessive evaporation of water to protect the DOPA@DNG stability as a skin patch. Evidence showed that the two-layer hydrogel has water retention. Water retention still remains at over 50% after keeping the hydrogel in air for 3 days. These properties mean the two-layer gel based on the DOPA/ZnO doped DN hydrogels could be used as a transdermal patch for numerous applications in drug delivery, wearable devices, and electronic skin.

Agar oligosaccharides: A review of preparation, structures, bioactivities and application.

Agar, a gelatinous polysaccharide which is in the cell wall of many red algae, is widely used as food and gelling agent. Agar oligosaccharides (AOs), the hydrolysate of agar, show much more kinds of bio-activities because of its lower molecular weight, better water solubility and higher absorption efficiency. It is indicated that AOs with different structure and degree of polymerization, i.e. series of agaro-oligosaccharides and neoagaro-oligosaccharides, can be obtained under different preparation conditions. In addition, the biological activities of AOs are diversely and closely correlated to the composition and structure. This review aims to comprehensively summarize the preparation, structural characteristics and bio-activities of AOs, so as to provide a reference for applications of AOs as marine natural products in pharmacological, cosmetics and nutraceutical fields.

Protection of agarwood essential oil aroma by nanocellulose-graft-polylactic acid.

Essential oil products are often volatile, and their aromas cannot be effectively preserved over long periods of time. In this study, nanocellulose crystals were modified, and an amphiphilic copolymer was prepared by ring-opening polymerisation to produce wall materials. A nanocellulose crystal-grafted polylactic acid copolymer was successfully synthesised and characterised using nuclear magnetic resonance spectrometry, Fourier transform infrared spectrometry, X-ray diffraction, and thermogravimetric analysis. Because of the amphiphilic properties of the synthesised copolymer, an agarwood essential oil nanoemulsion system was prepared. Using transmission electron microscopy and dynamic laser light scattering, the nanoemulsion was observed to have an apparent shell-core structure. The nanoemulsion was uniformly distributed, and the system had good stability. Finally, the electronic nose results showed that the nanocellulose crystal-grafted polylactic acid copolymer micelle effectively protected agarwood essential oil aromas.

Agarase cocktail from agar polysaccharide utilization loci converts homogenized Gelidium amansii into neoagarooligosaccharides.

Neoagarooligosaccharides (NAOs) are drawing more and more attention because of their numerous bioactivities, yet limited number of agarases impedes NAOs production from red algae. In this study, predicted agar polysaccharide utilization loci (agar-PUL) were firstly used as inventory for agarase. 6 agarases were identified from agar-PULs and two of them were successfully expressed and analyzed. Then enzyme cocktail (GH16-1:GH16-2:Aga50D = 2:1:1) was proved to have highest synergistic effect. Finally homogenization was applied to G. amansii and proved to be an efficient way to release agar from tissues. When liquid-to-solid ratio was 9 g/150 mL, homogenization time was 20 min, and enzyme cocktail loading was 150 U/150 mL, maximum NAOs production (90.2 mg per 9 g wet G. amansii) could be achieved. Enzyme supported one-step process (ESOP) proposed in study is environment-friendly, time saving, cost saving and none-destructive, therefore has a potential industrial application in red algae utilization.

Improvement of cadaverine production in whole cell system with baker's yeast for cofactor regeneration.

Cadaverine, 1,5-diaminopentane, is one of the most promising chemicals for biobased-polyamide production and it has been successfully produced up to molar concentration. Pyridoxal 5'-phosphate (PLP) is a critical cofactor for inducible lysine decarboxylase (CadA) and is required up to micromolar concentration level. Previously the regeneration of PLP in cadaverine bioconversion has been studied and salvage pathway pyridoxal kinase (PdxY) was successfully introduced; however, this system also required a continuous supply of adenosine 5'-triphosphate (ATP) for PLP regeneration from pyridoxal (PL) which add in cost. Herein, to improve the process further a method of ATP regeneration was established by applying baker's yeast with jhAY strain harboring CadA and PdxY, and demonstrated that providing a moderate amount of adenosine 5'-triphosphate (ATP) with the simple addition of baker's yeast could increase cadaverine production dramatically. After optimization of reaction conditions, such as PL, adenosine 5'-diphosphate, MgCl2, and phosphate buffer, we able to achieve high production (1740 mM, 87% yield) from 2 M L-lysine. Moreover, this approach could give averaged 80.4% of cadaverine yield after three times reactions with baker's yeast and jhAY strain. It is expected that baker's yeast could be applied to other reactions requiring an ATP regeneration system.

Maximizing the oil content in polysaccharide-based emulsion gels for the development of tissue mimicking phantoms.

Formulations based on agar and κ-carrageenan were investigated for the production of emulsion gels applicable as tissue mimicking phantoms. The effects of the polysaccharide matrix, the oil content and the presence of surfactants on the micro-/nanostructure, rheology, and mechanical and dielectric properties were investigated. Results showed a high capacity of the agar to stabilize oil droplets, producing gels with smaller (10-21 µm) and more uniform oil droplets. The addition of surfactants allowed increasing the oil content and reduced the gel strength and stiffness down to 57 % and 34 %, respectively. The permittivity and conductivity of the gels were reduced by increasing the oil content, especially in the agar gels (18.8 and 0.05 S/m, respectively), producing materials with dielectric properties similar to those of low-water content tissues. These results evidence the suitability of these polysaccharides to design a variety of tissue mimicking phantoms with a broad range of mechanical and dielectric properties.

Effectiveness of Cymbopogon citratus Oil Encapsulated in Chitosan on Colletotrichum gloeosporioides Isolated from Capsicum annuum.

One of the principal etiological agents associated with losses in horticultural crops is the fungus Colletotrichum sp. This study aimed to evaluate the in vitro effectiveness of the essential oil (EO) from Cymbopogon citratus in chitosan supports for the control of Colletotrichum gloeosporioides isolated from sweet pepper plants. METHODS: The extraction and phytochemical analysis of the EO of C. citratus were performed along with its encapsulation in chitosan-agar in order to compare it with other techniques and determine its effect on C. gloeosporioides. RESULTS: The EO from the citral chemotype (58%) encapsulated in the chitosan-agar, with an 83% encapsulation efficiency in mass percentage, resulted in the total inhibition of mycelial growth at a minimum inhibitory concentration of 1370 ppm. This concentration was effective in controlling the disease under greenhouse conditions. The effectivity of the capsules containing EO was superior to that of other controls using EO evaluated in vitro. The capsules demonstrated an effective period of 51 days, with an additional 30 days of effectiveness after a reinfection cycle, thus providing similar results to the control with Trichoderma sp. CONCLUSIONS: Chitosan capsules present a promising strategy in the use of C. citratus EO on C. gloeosporioides, and they are highly effective and stable under in vitro and field conditions.

Decorated Au NPs on agar modified Fe3O4 NPs: Investigation of its catalytic performance in the degradation of methylene orange, and anti-human breast carcinoma properties.

This work describes an eco-friendly approach for in situ immobilization of Au nanoparticles on the surface of Fe3O4 nanoparticles, with help of Agar and ultrasound irradiations, without using any toxic reducing and capping agents. The structure, morphology, and physicochemical properties were characterized by various analytical techniques such as Fourier transformed infrared spectroscopy (FT-IR), field emission scanning electron microscopy (FESEM), transmission electron microscopy (TEM), energy dispersive X-ray spectroscopy (EDS), X-ray diffraction (XRD), inductively coupled plasma (ICP) and vibrating sample magnetometer (VSM). The desired catalyst showed great efficiency in the reductive degradation of methylene orange (MO) dye over NaBH4 at room temperature. The MO was fully reduced in only 70 s and achieved rate constant of 9.6 × 10-2 s-1. The catalyst was reused for 10 runs without significant loss in catalytic activity. Cell viability of Fe3O4/agar/Au NPs was very low against breast adenocarcinoma (MCF7), breast carcinoma (Hs 578Bst), infiltrating ductal cell carcinoma (Hs 319.T), and metastatic carcinoma (MDA-MB-453) cell lines without any cytotoxicity on the normal cell line. According to the above findings, the Fe3O4/agar/Au NPs may be administrated for the treatment of several types of human breast carcinoma in humans.

An experimental aerosol air-agar interface mouse lymphoma assay methodology.

This work investigates a completely novel and experimental concept of exposing L5178Y cells at the air-agar-interface to mainstream cigarette smoke aerosol (Kentucky reference 3R4F). This study highlights the associated challenges of combining a suspension cell line alongside an in vitro aerosol exposure system. To achieve a monolayer, cells were 'seeded' in a concentrated cell super-mix suspension onto an RPMI/agar-matrix -base. The resulting cell suspension media was adsorbed into the agar base leaving the L5178Y cells lightly suspended on the agar surface, approximating a monolayer. Cells were deemed supportable on the agar-matrix, viable and recoverable. Using Vitrocell VC 10 exposure system and the Ames 4 exposure module, L5178Y cells were successfully exposed to a dynamic cigarette smoke aerosol, recovered and assessed for mutant frequencies, using standard assay procedures. Method development included assessment of flowing air conditions, plating efficiency and recovery of L5178Y cells from the agar-matrix surface. Positive controls MMS and B[a]P were successfully incorporated into the agar-matrix and metabolic activation was achieved by S-9 incorporation into the same agar-base-matrix. B[a]P demonstrated metabolic activation and positive response, suggesting a clear cellular interaction with the agar-matrix. Whole smoke exposed cells in the presence of metabolic activation showed a clear dose response and increasing mutant frequencies, well in excess of the controls (air and incubator) and the global evaluation factor following a 2 or 3 day expression period. This experimental concept demonstrates that L5178Y cells can be exposed to cigarette smoke aerosol, using a completely novel and a previously untested approach. Although this work successfully demonstrates the approach is viable and cells can be plated and maintained on an agar-matrix, more optimisation and robustness assessment is required before it can be considered fully adapted and used alongside other whole aerosol methodologies for the assessment of cigarette smoke and other inhaled aerosols.

Comparison of four agar media for the enumeration of the Mycobacterium abscessus complex.

Background: Traditional culture of nontuberculous mycobacteria (NTMs) has involved egg-based formulations (Lowenstein-Jensen medium) or defined media (Middlebrook formulations), which have disadvantages of composition complexity, availability, and cost. This study quantitatively compared three non-selective, non-blood based basal agars with Columbia blood agar (CBA), to enumerate Mycobacterium abscessus complex organisms in pure culture. Methods: M. abscessus subsp. massiliense, M. abscessus subsp. bolletii, and M. abscessus subsp. abscessus were employed. Inocula of each of these were counted on three basal agar media, including (i) standard plate count agar (SPCA), (ii) tryptone soya agar (TSA), and (iii) Mueller-Hinton agar (MHA) and compared to counts on CBA. Results: All NTM isolates of all subspecies grew successfully on all four media examined. The growth was most profuse on SPCA, with a mean colony diameter of 3 mm, whereas the mean colony diameter on all other media was 1 mm. Statistically, there was no significant difference in counts when comparing CBA with SPCA or MHA (P > 0.05), whereas there was a statistically significant difference between CBA and TSA (P = 0.01). There was no statistically significant difference between SPCA and MHA (P = 0.53). Conclusion: This study indicates that SPCA and MHA are equally effective as CBA, when enumerating of M. abscessus complex organisms. Employment of TSA gave significantly lower counts than CBA (P = 0.01) and therefore should not be employed when enumerating these organisms. SPCA yielded the most profuse growth of all media examined. In addition to these advantages, given that SPCA does (i) not require blood as a medium constituent, (ii) is simple to reconstitute, (iii) is relatively cheap, and (iv) is widely available commercially, this study endorses employment of SPCA for the nonselective culture of M. abscessus complex organisms, including enumeration.

Implications of agar and agarase in industrial applications of sustainable marine biomass.

Agar, a major component of the cell wall of red algae, is an interesting heteropolysaccharide containing an unusual sugar, 3,6-anhydro-L-galactose. It is widely used as a valuable material in various industrial and experimental applications due to its characteristic gelling and stabilizing properties. Agar-derived oligosaccharides or mono-sugars produced by various agarases have become a promising subject for research owing to their unique biological activities, including anti-obesity, anti-diabetic, immunomodulatory, anti-tumor, antioxidant, skin-whitening, skin-moisturizing, anti-fatigue, and anti-cariogenic activities. Agar is also considered as an alternative sustainable source of biomass for chemical feedstock and biofuel production to substitute for the fossil resource. In this review, we summarize various biochemically characterized agarases, which are useful for industrial applications, such as neoagarooligosaccharide or agarooligosaccharide production and saccharification of agar. Additionally, we succinctly discuss various recent studies that have been conducted to investigate the versatile biological activities of agar-derived saccharides and biofuel production from agar biomass. This review provides a basic framework for understanding the importance of agarases and agar-derived saccharides with broad applications in pharmaceutical, cosmetic, food, and bioenergy industries.

Distribution and Potential Effects of 17ß-Estradiol (E2) on Aeromonas Diversity in Wastewater and Fish Samples.

BACKGROUND AND OBJECTIVE: Recently, there has been evidence for the accumulation of steroid hormones in the water environment with negative consequences on fish and humans. However, there is paucity of information on how the steroid hormones influence the microbial community in environmental waters. The objective of this study was to determine the occurrence of 17ß-estradiol (E2) and its potential influence on the diversity of Aeromonas spp. MATERIALS AND METHODS: Wastewater samples were obtained from sewage treatment plants in northern South Africa and fish samples were collected from the Nandoni dam. Aeromonas spp. were isolated using microbiological methods and PCR protocols were used for their identification. A commercial Elisa kit was used for measuring the concentration of 17ß-estradiol (E2) from the wastewater samples as well as the fish samples. RESULTS: 17ß-estradiol (E2) was found in high concentration in sewage samples varying from 0.32-348.6 pg mL-1 while in fish samples, it ranged from 1.1-73.6 pg mL-1. There was a tendency of samples with high E2 concentrations to have higher diversity of Aeromonas spp., implying that steroid hormones may serve as nutrient for Aeromonas spp. Aeromonas hydrophila was the most prevalent species (71%), followed by A. sobria with (68%). CONCLUSION: The presence of Aeromonas spp. in environmental waters and fish that is consumed by the local community poses a serious health concern. The high content of E2 in treated wastewater is of serious concern as well. For the first time, the present study showed a positive impact of E2 on Aeromonas growth.