Expression and Characterization of a Novelλ-Carrageenase Cgl150A_Wa from Wenyingzhuangia aestuarii

λ-Carrageenan is a highly sulfated polysaccharide alternating of 1,4-O-α-D-galactopyranose-2,6-sulfate(D2S,6S)and 1,3-O-β-D-galactopyranose-2-sulfate(G2S).λ-Carrageenases are desirable tools forλ-carrageenan degradation.Based on the genome mining,a novelλ-carrageenase Cgl150A_Wa was cloned from the bacterium Wenyingzhuangia aestuarii and expressed in Escherichia coli.Cgl150A_Wa was an endo-acting enzyme and exhibited its maximum activity at 30℃and pH 8.0.By employing a glycomics strategy that combined ultra-performance liquid chromatography-mass spectrometry analysis and glycoinformatics,Cgl150A_Wa was proven to degradeλ-carrageenan octaose and hexaose,and the major hydrolysis product of Cgl150A_Wa wasλ-carrageenan tetrose.In addition to the typicalλ-carrageenan motifs,the active center of Cgl150A_Wa might tolerate desulfatedλ-carrageenan motifs.Cgl150A_Wa is a potential biotechnological tool for preparingλ-carrageenan oligosaccharides and structural investigation.

Analgesic and anti-inflammatory activity of Cotinus coggygria Scop.extracts in vivo

Objective:To assess the analgesic and anti-inflammatory effects of standardized extract of Cotinus coggygria(C.coggygria)in different animal models.Methods:C.coggygria extracts(25,50,and 100 mg/kg)were administered to rats and mice(n=6)during hot plate,tail-flick,acetic acid-induced writhing,and formalin tests to determine its analgesic efficacy.The anti-inflammatory activity of C.coggygria extracts was evaluated by histamine and carrageenan-induced paw edema,cotton pellet-induced granuloma,and acetic acid-induced peritoneal capillary dye leakage tests.Results:C.coggygria extracts(50 and 100 mg/kg)significantly alleviated thermal and chemical-induced pain in rodents(P<0.05).It also demonstrated notable anti-inflammatory properties by mitigating histamine and carrageenan-induced paw edema,granuloma deposits,and vascular permeability(P<0.05).Moreover,C.coggygria extracts remarkably reduced TNF-α,IL-1β,IL-6,COX-2,and oxidative stress in rat paws(P<0.05).Carrageenan-induced histological aberrations in hind paw tissues were effectively(P<0.05)mitigated by treatment with C.coggygria extracts.Conclusions:C.coggygria Scop.extracts show analgesic and anti-inflammatory effects via inhibition of COX-2 and inflammatory and oxidative mediators.

Carrageenan Fiber Prepared by a New Process Route of Ba^(2+)Ion Pre-Crosslinking in the Spinning Solution

Ba^(2+)pre-crosslinked carrageenan fiber(Ba/CAF)was prepared by adding a small amount of Ba^(2+) to the carrageenan(CA)solution as the spinning solution.Ba/CAF-n/A,Ba/CAF-n/B and Ba/CAF-n/C were prepared with ethanol solution(combine A),high concentration BaCl_(2)solution(combine B)and low concentration BaCl_(2)solution(combine C),as coagulation bath and stretch bath,respectively.The combination of coagulation bath and stretch bath suitable for Ba^(2+) pre-crosslinking wet spinning was screened.The results showed that Ba^(2+) can induce the birefringence of the CA molecular chain,and the Ba^(2+) pre-crosslinking effect is the best when the CA mass fraction is 8.0 wt%.From the perspective of production safety,fiber performance and spinning cost,the coagulation bath of 3.5 wt%BaCl_(2)solution and stretch bath of 1.7 wt%BaCl_(2)solution,that is,combination C with low concentration BaCl_(2)solution,is the best choice.Ba/CAF-8.0/C was obtained under the best conditions.The linear intensity,water absorption and flame retardancy study showed that the breaking strength of Ba/CAF-8.0/C is as high as 1.61 cN/dtex,the water absorption was 649.2%and 574.3%,in deionized water and normal saline,respectively,and the LOI value reached 32.

A Comparative Analysis of Four Kinds of Polysaccharides Purified from Furcellaria lumbricalis

Four kinds of polysaccharides (FA, FB, FC and FD) were extracted and purified from a red alga Furcellaria lumbri-calis. Their monosaccharide composition, average molecular mass (Mw) and structural feature were determined by gas chromatog-raphy (GC), high performance gel permeation chromatography (HPGPC) and Fourier transform infrared spectroscopy (FTIR), respectively. The results showed that FA and FB were composed of galactose (Gal), 3,6-anhydro-galactose (AnG) and 4-sulfated-galactose (G4S), which were structurally similar to β-carrageenan and κ-carrageenan. FC was composed of sulfated galactose and small amount of AnG, indicating that it was a hybrid-sulfated galactan. FD was composed of α-1, 4-glucose and belonged to floridean starch. The respective yields of FA, FB, FC and FD were 4.84%,33.74%,8.76% and 13.5%, and the cor-responding Mw were 537 kD, 432 kD, 513 kD and 51 kD respectively. Owning to its high content ( >60%) of polysaccharide, F. lumbricalis will be a good source of marine sulfated polysaccharide and shows its potential utilization in food and medicine.

Analgesic and anti-inflammatory activities of Passiflora foetida L.

Objective:To investigate the analgesic and anti-inflammatory activities of ethanol extract of Passiflora foetida(P.foetida) leaves.Methods:Ethanol extract of P.foetida leaf was evaluated for analgesic action by acetic acid-induced writhing and hot plate method in albino mice.The anti-inflammatory property of ethanolic leaf extract was tested by carrageenan induced acute paw edema and histamine induced acute paw edema in rats.Results:The dose 200 mg/kg of P. foetida leaf extract exhibited highest significant analgesic activity[(13.50±0.43) min]at a reaction time of 20 min in hot plate method in mice.The ethanol extract of leaf dose 100 mg/kg produced a highly significant anti inflammatory effect[(1.302±0.079) mL]in rats.Conclusions:It is very clear that P.foetida also has analgesic and anti-inflammatory activities for the pharmaceuticals.

Anti-inflammatory activity of the leaf extacts of Gendarussa vulgaris Nees

Objective:To evaluate the anti-inflammatory property of the leaf exacts of Gendarussa vulgaris(G.vulgarix) Nees.Methods:G.vulgarix Nees of the family Apocynaceae is a medium sized tree grown in semishade or no shade and is common in the Ernad and Nilambur taluks of Kerala. Various parts of this plant have been used in the treatment of ulcers,sores,inflammation, dyspepsia,healing of wounds,etc.The present study aimed at the evaluation of anti-inflammatory property of the aqueous and alcoholic extracts of the leaves by both in vitro and in vivo methods. In vitro method was estimated by human red blood cell membrane stabilisation(HRBC) method and in vivo method was estimated on the carrageenan induced paw oedima.Results:Both the methods showed significant anti-inflammatory property of the different extracts tested. Conclusions:The alcoholic extract at a concentration of 300 mg/mL showed potent activity on comparing with the standard drug diclofenac sodium.

Anti-inflammatory activity of the glandular extracts of Thunnus alalunga

Objective:To evaluate the anti-inflamattory activity of Thunnus alalunga by both in vitro and in vivo methods.Methods:Anti-inflammatory activity of the chloroform water extract of Thunnus alalunga was done by both in vitro and in vivo methods.In vitro method was done by human red blood cells membrane stabilization method(HRBC).In vivo evaluation was estimated on Wister albino rats.Acute toxicity studies were done on the extract and no toxicity was reported.Results: The percentage protection exhibited by 300 mg/mL concentration was more when compared to the other ones.The 400 mg/mL concentration showed potent activity on comparison with the standard during in vivo evaluation.Conclusions:In both means of estimation the extract of Thunnus alalunga was found to possess significant anti-inflammatory activity.

Immobilization of microorganisms using carrageenan gels coated with chitosan and application to biodegradation of 4-chlorophenol

A new cell immobilization method based on the replacement of KCl by KCl+chitosan as the gelling agent was developed. The experimental results showed that through addition of chitosan into gelling agent, the mechanical strength and the thermal stability of the carrageenan gel were greatly improved. The new immobilization method was used to entrap a chlorophenol degrading microorganism. The immobilized microbial cells were applied for chlorophenol biodegradation. The experiments demonstrated that immobilized cells exhibit a higher bioactivity in the degradation of chlorophenol than free cells.

Evaluation of Analgesic and Anti-Inflammatory Activities on Ethanolic Extract of <i>Terminalia chebula</i>Fruits in Experimental Animal Models

The present study was undertaken to evaluate the analgesic and anti-inflammatory activities of ethanolic extract of Terminalia chebula (commonly known as Haritaki) fruits in experimental animal models. The study was carried out using Swiss Albino mice (20-25 g) and Long Evans rats (100-150 g) of either sex. The analgesic activity of Terminalia chebula was assessed by using hot plate method. For the determination of analgesic effect, doses of ethanolic extract of Terminalia chebula used in the present study were 250 mg/kg and 500 mg/kg body weight (BW). Anti-inflammatory effect was analyzed by carrageenan induced paw edema method with the administration dose of 300 mg/kg BW of animals. The analysis of experimental data was performed by statistical process of ANOVA to determine the variability of sample, while Dunnet’s test was performed for evaluation of comparative analgesic and anti-inflammatory activity of Terminalia chebula with control and standard. The animals were divided into four treatment groups of six animals each and the “Mean ± SEM” is the statistical identifiable value of the data and P values <0.05 was considered statistically significant. Hot plate test showed a significant increase in the mean reaction time to heat stimuli in hot plate method at both 250 mg/kg and 500 mg/kg BW doses throughout the observation period in 30 minutes and 60 minutes after treatment, which was comparable to the standard ketorolac and control group. In carrageenan induced paw edema method, considerable results were found after determining the percentage change in paw volume in extract. In both cases of analgesic and anti-inflammatory study, % inhibition of pain and inflammation were evaluated. Comparing with control, largest inhibition was found in inhibiting inflammation 5 hours after treatment, while the largest inhibition of pain was obtained in 30 minutes and 60 minutes after treatment of doses. The present study suggests that ethanolic extract of Terminalia chebula fruits has significant analgesic and anti-inflammatory activities.

Utilization of Different Hydrocolloid Combinations in Gluten-Free Bread Making

Coeliac disease (CD) is a common lifelong disorder with small bowel malabsorption and gluten intolerance. The only effective treatment for CD is a strict lifelong renunciation of gluten-containing foods. Currently, the cereal products designed to meet the requirements of CD patients, especially gluten-free bread, are scarce, of low quality and poor flavor. Therefore, preparation of high quality and affordable bread for coeliac patients was the main objective of this study. Three kinds of gluten-free bread were prepared, in which wheat flour was substituted with rice and corn flour at a ratio of 5:1, respectively. Gluten was substituted using different hydrocolloids: 1% xanthan and 1% carrageenan (Mix A), GFB A;1% xanthan and 1% pectin (Mix B), GFB B;and 1% carrageenan and 1% pectin (Mix C), GFB C. These three types of bread were compared to control bread produced from wheat flour (WB). Sponge and dough method with some modifications was used to produce the bread samples with a developed formula (0.8% yeast, 4.0% sugar, 2% salt, 7.0% shortening, 2.0% non-fat dry milk, and 1% sodium stearoyl lactylate (SSL)). Determination of the chemical composition, rheological properties and physical characteristics, as well as sensory evaluation of the bread samples, was undertaken. Results showed that the moisture content of WB bread (control) was significantly lower than that of all types of gluten-free bread, and there was no significant difference in ash, protein, and lipid contents among the gluten-free bread samples. GFB A, GFB B and GFB C displayed no significant differences in their specific volumes. The sensory evaluation showed that GFB C received the lowest sensory evaluation score. Therefore, GFB A and GFB B were the best gluten-free samples.

De novo sequencing and comparative analysis of three red algal species of Family Solieriaceae to discover putative genes associated with carrageenan biosysthesis

Betaphycus gelatinus, Kappaphycus alvarezii and Eucheuma denticulatum of Family Solieriaceae, Order Gi-gartinales, Class Rhodophyceae are three important carrageenan-producing red algal species, which pro-duce different types of carrageenans, beta （β）-carrageenan, kappa （κ）-carrageenan and iota （ι）-carrageenan. So far the carrageenan biosynthesis pathway is not fully understood and few information is about the So-lieriaceae genome and transcriptome sequence. Here, we performed the de novo transcriptome sequencing, assembly, functional annotation and comparative analysis of these three commercial-valuable species using an Illumina short-sequencing platform Hiseq 2000 and bioinformatic software. Furthermore, we compared the different expression of some unigenes involved in some pathways relevant to carrageenan biosynthe-sis. We finally found 861 different expressed KEGG orthologs which contained a glycolysis/gluconeogenesis pathway （21 orthologs）, carbon fixation in photosynthetic organisms （16 orthologs）, galactose metabolism （5 orthologs）, and fructose and mannose metabolism （9 orthologs） which are parts of the carbohydrate me-tabolism. We also found 8 different expressed KEGG orthologs for sulfur metabolism which might be impor-tantly related to biosynthesis of different types of carrageenans. The results presented in this study provided valuable resources for functional genomics annotation and investigation of mechanisms underlying the biosynthesis of carrageenan in Family Solieriaceae.

Optimization of genistein solubilization byκ-carrageenan hydrogel using response surface methodology

κ-Carrageenan was explored to improve genistein solubility by matrix retention.The corresponding maximum value in the efficiency of retention(Re)(50.48 mg/100 mg)was achieved when variables were set as:pH 4.76,temperature 52.12◦C and genistein concentration 0.27 mg/mL.The coefficient of determination(R2)of the response surface regression model presented in this study was 0.9848.The evidences from XRD,DSC and FT-IR attested the amorphous form of genistein in hydrogel matrix.Importantly,the solubility of genistein in hydrogel amorphous form(16.84–34.42g/mL)was much higher than that of its free crystalline form(1.89–6.09g/mL)over 30–90◦C.©2013 Beijing Academy of Food Sciences.Production and hosting by Elsevier B.V.All rights reserved.

Impact of Ultra-Sonication on Gelation Process in Dairy Products Incorporated with Polysaccharides

Conventional or traditional methods for the use of food processing have been outperformed by novel approaches recently. One such promising technology is ultra-sonication and recent studies have shown better effects in physical characteristics of food which are induced by ultrasound. The previous researches further emphasize that the introduction of ultra-sonication on dairy proteins has resulted in the improvement of their structural, physical and chemical properties while it is widely used in hydrocolloid systems to accelerate the gelation process. Therefore this theory could be useful for the gelation of milk proteins in the presence of a polysaccharide. After discussing briefly on hydrocolloids and the theory of ultrasound, the possibility and the impact of ultrasound on gelation of milk protein mixtures with Carrageenan is highlighted in this study. The inherent characteristics of both milk proteins and carrageenan and properties of the ultrasound treatment have cumulatively effect on gelation. Therefore, there is a necessity to develop specific variables depending on the type of dairy product.

Formulation Development of a Carrageenan Based Delivery System for Buccal Drug Delivery Using Ibuprofen as a Model Drug

Solvent cast films are used as oral strips with potential to adhere to the mucosal surface, hydrate and deliver drugs across the buccal membrane. The objective of this study was the formulation development of bioadhesive films with optimum drug loading for buccal delivery. Films prepared from κ-carrageenan, poloxamer and polyethylene glycol or glycerol, were loaded with ibuprofen as a model water insoluble drug. The films were characterized using texture analysis (TA), hot stage microscopy (HSM), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), scanning electron microscopy (SEM), x-ray powder diffraction (XRPD), high performance liquid chromatography (HPLC) and in vitro drug dissolution. Optimized films were obtained from aqueous gels containing 2.5% w/w κ-carrageenan 911, 4% w/w poloxamer 407 and polyethylene glycol (PEG) 600 [5.5% w/w (non-drug loaded) and 6.5% w/w (drug loaded)]. A maximum of 0.8% w/w ibuprofen could be incorporated into the gels to obtain films with optimum characteristics. Texture analysis confirmed that optimum film flexibility was achieved from 5.5% w/w and 6.5% (w/w) of PEG 600 for blank films and ibuprofen loaded films respectively. TGA showed residual water content of the films as approximately 5%. DSC revealed a Tg for ibuprofen at -53.87。C, a unified Tm for PEG 600/poloxamer mixture at 32.74。C and the existence of ibuprofen in amorphous form, and confirmed by XRPD. Drug dissolution at a pH simulating that of saliva showed that amorphous ibuprofen was released from the films at a faster rate than the pure crystalline drug. The results show successful design of a carrageenan and poloxamer based drug delivery system with potential for buccal drug delivery and showed the conversion of crystalline ibuprofen to the amorphous form during film formation.

Evaluation of anti-inflammatory potential of leaf extracts of Skimmia anquetilia

Objective:To evaluate anti-inflammatory potential of leaf extract of Skimmia anquetilia by invitro and in-vivo anti-inflammatory models.Methods:Acute toxicity study was carried out to determine the toxicity level of different extract using acute toxic class method as described in Organization of Economic Co-operation and Development Guidelines No.423.Carrageenan(1% w/w) was administered and inflammation was induced in rat paw.The leaf extracts of Skimmia anquetilia were evaluated for anti-inflammatory activity by in-vitro human red blood cell(HRBC) membrane stabilization method and in-vivo carrangeenan-induced rat paw edema method. Results:The in-vitro membrane stabilizing test showed petroleum ether(PE),chloroform(CE), ethyl acetate(EE),methanol(ME) and aqueous extracts(AE) showed 49.44%,59.39%,60.15%,68.40% and 52.18%protection,respectively as compared to control groups.The in-vivo results of CE,EE and ME showed 58.20%.60.17%and 67.53%inhibition of inflammation after 6h administration of test drugs in albino rats.The potency of the leaf extracts of Skimmia anquetilia were compared with standard diclofenac(10 mg/kg) which showed 74.18%protection in in-vitro HRBC membrane stabilization test and 71.64%inhibition in in-vivo carrangeenan-induced rat paw edema model. The ME showed a dose dependent significant(P< 0.01) anti-inflammatory activity in human red blood cell membrane stabilization test and reduction of edema in carrageenan induced rat paw edema.Condusioiis:The present investigation has confirmed the anti-inflammatory activity of Skimmia anquetilia due to presence of bioactive phytoconstitutes for the first lime and provide the pharmacological evidence in favor of traditional claim of Skimmia anquetilia as an antiinflammatory agent.

The anti-nociceptive effect of bufalin,an active ingredient from toad venom via modulating voltage-gated sodium channels

OBJECTIVE Toad venom(Venenum Bufonis)isalways used for analgesia in China from ancient to modern times,but the effective component of it remains unclear.METHODS In the present study,we investigated the anti-nociceptive effect and the underlying mechanism ofbufalin,an active ingredient fromtoad venom by animal behavior,patch clamp and calcium imaging.RESULTS Bufalin could significantly relieve formalin-induced spontaneous flinching and licking response as well as carrageenan-induced mechanical and thermal hyperalgesia.Using the whole-cel patch-clamp recording,bufalincaused remarkable suppressive effect on the peak currents of Na+channels in dorsal root ganglion neuroblastoma ND7-23 cel line in a U-shaped dependent manner.In addition,bufalinprompted the voltage-dependent activationand caused a negative shift of the fast-state inactivation of Na+channels.However,bufalin produced insignificant effect not onlyon voltage-dependent Kv4.2,Kv4.3 and BK channels,but also on the capsaicin induced Ca2+influx.CONCLUSION The present results indicate bufalin is capable of producing remarkable anti-nociceptive effects whichis probably ascribed to its specific modulation of voltage-gated Na+channels.

Effects of tetrandrine on experimental pleurisy in rats

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Apparent Viscosity of a Skim Milk Based Dessert: Optimization Through Response Surface Methodology

In this study, the apparent viscosity of a skim milk based dessert was optimized through the use of a 5-level Central Composite Design. The effect of carrageenan, guar gum, sucrose and pH on the dessert's apparent viscosity was esti-mated. The dessert's flow behavior was also analyzed. A rotational viscometer was used for the the apparent viscosity measurements. All formulations presented a non-Newtonian, shear-thinning behavior. Except for carrageenan concentration, all of the design factors affected apparent viscosity, being the effects better described by second order polynomial models. The following combinations of the variable ingredients yielded the best apparent viscosity: guar gum concentration: 0.0-0.315% (w/w) and sucrose concentration: 5.000-8.615% (w/w);or guar gum concentration: 0.109-0.315% (w/w) and sucrose concentration: 5.000-11.000% (w/w);in both cases, carrageenan and pH were kept at 0.020% (w/w) and 6.50, respectively.

Dietary Supplementation of <i>κ</i>-Carrageenan to Improve the Physio-Chemical and Functional Properties of White Bread

Fiber intake improves gut health and prevents non-communicable diseases. The current study investigates the substitution of carrageenan in white bread and evaluates its effect on the physiochemical and structural characteristics of bread. The 100% wheat flour was used as control and the test sample contained 4% carrageenan. The physio-chemical analysis showed that carrageenan-substitution improved the hydration properties of the flour (WHC—1.33 g/g;SC—3.50 ml/g). Carrageen substituted bread had reduced the loaf volume. The fiber content in carrageenan-substituted bread was noticeably higher (9.4 g%) than control (3.5 g%). Crude lipid (4.6 g%) and protein (7.0 g%) content improved with carrageenan-substitution. The mineral contents (Na, K, Mg, Ca, Fe, and Zn) were increased in carrageenan-breads. The texture profile analysis showed a decreased hardness (H1—92.3 N, H2—62.5 N) and improved springiness (5.3 mm) in carrageenan-bread.

Preparation and Properties of Carrageenan-g-Poly(Acrylic Acid)/Bentonite Superabsorbent Composite

A novel biopolymer-based superabsorbent hydrogel composite based on kappa-carrageenan (κC) have been prepared via graft copolymerization of acrylic acid (AA) in the presence of bentonite powder using methylenebisacrylamide (MBA) as a crosslinking agent and ammonium persulfate (APS) as an initiator. The hydrogel structure was confirmed using FTIR spectroscopy and the morphology of the samples was examined by scanning electron microscopy (SEM). The affecting variables onto graft polymerization (i.e. AA, MBA and APS concentration, as well as the bentonite amount) were systematically optimized to achieve a hydrogel with swelling capacity as high as possible. The results of Brunauer–Emmett–Teller (BET) analysis showed that the average pore diameter of the synthesized hydrogel was 11.5 nm. The effect of various salt media and solutions with different pHs on the swelling of the superabsorbent was also studied.