Rooster Comb Research Articles

Protecting the Environment from Waste. Development of a New Drug Delivery Biopolymer for Obtaining Hyaluronic Acid from Rooster Comb

Protecting the Environment from Waste. Development of a New Drug Delivery Biopolymer for Obtaining Hyaluronic Acid from Rooster Comb

Microbial hyaluronic acid production in the 21 century: a roadmap toward high production, tailored molecular weight

Hyaluronic acid is a biopolymer that has gained attention from both researchers and the public since its discovery in 1934. In 2022, its global annual demand was 667 MT, with a market value of 9.4 billion dollars and an expected compound growth rate of 7.7% from 2024 and 2030. Such popularity lies in its many applications, which ranges from pharmaceutical to Tissue Engineering. Its molecular weight (MW) determines which application hyaluronic acid is better suited for: high MW (over 1 MDa) is used in medical applications such as osteoarthritis and ophthalmology, and low MW (less than 0.05 MDa) are suited for moisturizing cosmeceuticals. This makes hyaluronic acid molecular weight an important parameter for its production. The first industrial process was based on extraction from rooster combs. However, it shifted toward microbial production with Streptococcus zooepidemicus. Studies focus on improving its production and tailoring its molecular weight by accessing optimum temperature, pH, aeration, agitation, carbon, and nitrogen sources concentration and origin. his review aims to provide an overview of the current knowledge on the effects of metabolic and environmental factors in microbial hyaluronic acid production. The review shows that there has been extensive research in optimizing cultivation medium composition, temperature, pH, and aeration between 2018 and 2023, but further studies are required to optimize the metabolism of S. zooepidemicus towards maximum biopolymer production and molecular weight.

Effect of Online Simulation-Based Education on Nurses' Knowledge and Skills Toward Electrocardiogram Interpretation

Background: Osteoarthritis (OA); a whole joint disorder is a most common and progressive disease that worsens with age. Synovitis and synovial effusion can play important role in early OA diagnosis and progression. Limited pharmacological interventions affecting synovial effusion are studied clinically in Indian population. Aim: This was a pilot, observational, single arm, open label study in real world set-up designed to evaluate the efficacy and safety of Mobileye® capsules containing sodium hyaluronate along with collagen and polysaccharides derived from rooster comb extract in subjects suffering from synovial effusion and knee pain of osteoarthritis. Materials and Method: The study was conducted on total 30 subjects at Outpatient Department (OPD) at three different sites under the supervision of medical doctor. All subjects consumed Mobileye® capsule orally twice daily for 3 months. Synovial effusion, VAS score, WOMAC score and safety parameters were assessed as scheduled. Results: Treatment with Mobileye® capsule showed significant and progressive improvement in all study related parameters. Baseline score vs. day 90 score (mean + SD) for synovial effusion, WOMAC pain score, WOMAC stiffness score, WOMAC physical function score, WOMAC total score and VAS score were 8.52+2.960 mm vs. 2.58+0.903 mm (p<0.0001), 15.13+3.10 vs. 4.13+2.4 (p<0.0001), 6.63+1.098 vs. 1.60+1.192 (p<0.0001), 54.33+8.39 vs. 14.23+8.67 (p<0.0001), 76.13+ 11 vs. 20.33+ 10.86 and 8.13 + 1.28 vs. 1.93+1.17 (p<0.0001) respectively. No significant adverse events were reported. Conclusion: Oral intake of Mobileye® capsule is safe and effective intervention in reducing synovial effusion, pain, stiffness and in improving joint mobility. Keywords: Electrocardiogram interpretation, nurses' knowledge and skills, online simulation education.

A Novel Hyaluronic Acid Matrix Ingredient with Regenerative, Anti-Aging and Antioxidant Capacity.

Hyaluronic acid (HA) and proteoglycans (such as dermatan sulphate (DS) and chondroitin sulphate (CS)) are the main components of the extracellular matrix of the skin, along with collagen and elastin. These components decrease with age, which implies a loss of skin moisture causing wrinkles, sagging and aging. Currently, the external and internal administration of effective ingredients that can reach the epidermis and dermis is the main alternative for combating skin aging. The objective of this work was to extract, characterise and evaluate the potential of an HA matrix ingredient to support anti-aging. The HA matrix was isolated and purified from rooster comb and characterised physicochemically and molecularly. In addition, its regenerative, anti-aging and antioxidant potential and intestinal absorption were evaluated. The results show that the HA matrix is composed of 67% HA, with an average molecular weight of 1.3 MDa; 12% sulphated glycosaminoglycans, including DS and CS; 17% protein, including collagen (10.4%); and water. The in vitro evaluation of the HA matrix's biological activity showed regenerative properties in both fibroblasts and keratinocytes, as well as moisturising, anti-aging and antioxidant effects. Furthermore, the results suggest that the HA matrix could be absorbed in the intestine, implying a potential oral as well as topical use for skin care, either as an ingredient in a nutraceutical or a cosmetic product.

Hyaluronic Acid – from Production to Application: A Review

Hyaluronic acid (HA) was traditionally obtained by extraction from animal sources, specifically from rooster combs. However, this method of obtaining it has several disadvantages. Therefore, the number of researches has increased, and several microbial strains have been studied to synthesize HA. In this review, the characteristics of hearing aids were addressed; the microorganisms involved in the production of HA, operations for HA recovery and purification, and HA applications. Several species of microorganisms are capable of producing HA, mainly Streptococcus zooepidemicus, which can produce up to 7 g/L under ideal growing conditions. The culture medium, as well as the environmental conditions (pH, temperature, aeration, agitation), are factors that directly influence the production of HA, which can reduce or maximize it. Regarding recovery and purification methods, several techniques in sequence are used involving the precipitation of HA from the fermentation broth, usually using repeatedly organic solvents, surfactants, centrifugation, and membrane separation, among others. The functions of the HA cover several areas such as pharmaceutical, medical, and aesthetics, among others, causing the commercial demand for this biopolymer to increase every year, justifying research involving its bioproduction.

Effects of rooster comb degradation product containing low-molecular hyaluronic acid (INJUV) in individuals with eye dryness; open-label tr.ial with no control group.

Effects of rooster comb degradation product containing low-molecular hyaluronic acid (INJUV) in individuals with eye dryness; open-label tr.ial with no control group.

Preparation and characterization of decellularized rooster comb as a scaffold for tissue engineering applications

Decellularization is a method that has been widely used in tissue engineering especially in the last 20 years. In this study decellularized rooster comb was prepared and characterized for using it as a tissue scaffold. Treatment of tissues with sodium dodecyl sulfate (SDS) and Triton X-100 as two decellularization procedures in different time points were compared according to different parameters such as cytocompatibility, cell removal, preservation of extracellular matrix (ECM), and mechanical properties to find the optimum technique. Even though there was no difference regarding to efficiency on cell removal, SDS demonstrated better results on protection of tissue morphology in comparison with Triton X-100. Therefore, in general the samples treated with SDS showed higher levels of mechanical properties and cytocompatibility in comparison with Triton X-100 applied tissues. In the cuisines of many countries, rooster comb is discarded as a waste material however, in this study it was demonstrated that decellularized rooster comb could be utilized as a cheap, easily obtainable, and biocompatible scaffold. In conclusion, it was revealed that decellularized rooster comb is a promising biomaterial for using as scaffold and it is expected to be utilized for the further studies in particular on skin tissue engineering.

Repetitive Synthesis of High-Molecular-Weight Hyaluronic Acid with Immobilized Enzyme Cascades.

Industrial hyaluronic acid (HA) production comprises either fermentation with Streptococcus strains or extraction from rooster combs. The hard‐to‐control product quality is an obstacle to these processes. Enzymatic syntheses of HA were developed to produce high‐molecular‐weight HA with low dispersity. To facilitate enzyme recovery and biocatalyst re‐use, here the immobilization of cascade enzymes onto magnetic beads was used for the synthesis of uridine‐5’‐diphosphate‐α‐d‐N‐acetyl‐glucosamine (UDP‐GlcNAc), UDP‐glucuronic acid (UDP‐GlcA), and HA. The combination of six enzymes in the UDP‐sugar cascades with integrated adenosine‐5'‐triphosphate‐regeneration reached yields between 60 and 100 % for 5 repetitive batches, proving the productivity. Immobilized HA synthase from Pasteurella multocida produced HA in repetitive batches for three days. Combining all seven immobilized enzymes in a one‐pot synthesis, HA production was demonstrated for three days with a HA concentration of up to 0.37 g L−1, an average M W of 2.7–3.6 MDa, and a dispersity of 1.02–1.03.

Improving physical and biological properties of nylon monofilament as suture by Chitosan/Hyaluronic acid

One way to give some properties such as antibacterial and good frictional properties to sutures is the application of natural antibacterial and hydrophilic components on their surfaces through layer by layer assembly (LBL) technique. In this regard, Chitosan as an antibacterial polycationic natural polymer along with Hyaluronic acid (HA) as a polyanionic polysaccharide could be used to form a polyelectrolyte complex. In this study, HA was extracted from rooster comb using different solvents. Characterization of the extracted HA by FTIR and GPC analysis showed extracted HA with Mw = 2.53 × 105 Da had no cytotoxicity. Then, a nylon monofilament (NMy) was coated by the extracted HA and chitosan with different concentrations using bilayer coating technique. Two dyes also were loaded to coating layer to investigate the release behavior of these two drug models. The morphology of coated layer showed that coating NMy by chitosan (4% w/v) following by HA (8% w/v) with roughness of 164 ± 129 nm and friction coefficient of 0.26 had suitable interaction between two layers to prevent from exfoliation of coating layers. The antibacterial activity and controlled release of coated NMy indicated how a NMy coated by Chitosan and HA is a promising material for using as a suture.

Bioengineered production of glycosaminoglycans and their analogues.

Glycosaminoglycans (GAGs) are a class of linear polysaccharides, consisting of alternating disaccharide sequences of uronic acid and hexosamines (or galactose) with and without sulfation. They can interact with various proteins, such as growth factors, receptors and cell adhesion molecules, endowing these with various biological and pharmacological activities. Such activities make GAGs useful in health care products and medicines. Currently, all GAGs, with the exception of hyaluronan, are produced by extraction from animal tissues. However, limited availability, poor control of animal tissues, impurities, viruses, prions, endotoxins, contamination and other problems have increased the interest in new approaches for GAG production. These new approaches include GAGs production by chemical synthesis, chemoenzymatic synthesis and metabolic engineering. One chemically synthesized heparin pentasaccharide, fondaparinux sodium, is in clinical use. Mostly, hyaluronan today is prepared by microbial fermentation, largely replacing hyaluronan from rooster comb. The recent gram scale chemoenzymatic synthesis of a heparin dodecasaccharide suggests its potential to replace currently used animal-sourced low molecular weight heparin (LMWH). Despite these considerable successes, such high-tech approaches still cannot meet worldwide demands for GAGs. This review gives a brief introduction on the manufacturing of unfractionated and low molecular weight heparins, the chemical synthesis and chemoenzymatic synthesis of GAGs and focuses on the progress in the bioengineered preparation of GAGs, particularly heparin.

Biocompatibility study of different hyaluronan products for intra-articular treatment of knee osteoarthritis

BackgroundIntra-articular (IA) injection of hyaluronic acid (HA) (IA-HA) is a well-recognized treatment option for pain associated with symptomatic knee osteoarthritis (OA). IA-HA products differ in their HA content, molecular weight, cross-linking, and source of HA. These differences are assumed to affect the biocompatibility of the IA-HA products once injected inside the knee joint.MethodsIn the present study, we investigated the biocompatibility of three multiple-injection IA-HA products available in the global market. These included SUPARTZ FX™, a medium range molecular weight HA derived from rooster comb (Avian-HA); ORTHOVISC®, a high range molecular weight HA obtained through biological fermentation (Bio-HA); and SYNVISC®, a high molecular weight cross-linked hyaluronan derived from rooster comb (Avian-CL-HA). Rabbit knee joint tissues were histologically and biochemically examined after IA injection of the products. Furthermore, we compared the amounts of impurities in the IA-HA products.ResultsIA injection of Avian-CL-HA into rabbit knee joints induced the aggregation of inflammatory cells, infiltration of eosinophils, and an increase in the number of cells in the synovial fluid. However, these effects were not seen in the Avian-HA and Bio-HA groups. The residual protein content and the contaminant levels of bacterial endotoxins were below the limit of quantitation in all HA products. Avian-CL-HA contained relatively a large amount of (1 → 3)-β-D-glucan, but this was below the lower limit of quantification in the other HA products.ConclusionsThe present results clearly demonstrate that the biocompatibility of Avian-HA is comparable to that of Bio-HA, and they were both considered to have a favorable safety profile for the treatment of symptomatic OA of the knee. However, immunostimulatory activity was observed after injection of Avian-CL-HA: this might be a result of its unique cross-linking structure and/or the considerable amount of (1 → 3)-β-D-glucan impurity present in the formulation.

Extraction, purification and characterization of hyaluronic acid from Rooster comb

Hyaluronic acid, is extract by different procedures from various sources like pig, rabbit, oxes and human are available, but these processes have certain imitations like low yield, and also it requires the killing of these animals which is against the experimental ethics. In the present study, we have carried out the extraction of hyaluronic acid from cock’s comb which was further analyzed with qualitative test, viscosity, UV absorption, endotoxin detection assay. Also, the protein contamination of extracted hyaluronic acid was determined by using SDS-PAGE of hyaluronic acid was studied for checking the protein contaminants and it was noted that there were no bands observed in the well loaded with extracted hyaluronic acid sample indicating that the final extract of hyaluronic acid is not contaminated with the protein. The extraction and purification of hyaluronic acid by using the method reported here give pure hyaluronic acid. The viscosity of extracted hyaluronic acid was found to be 2.55 poise which is economical and can be used for industrial production of hyaluronic acid having clinical applications

Real-time monitoring of hyaluronic acid fermentation by in situ transflectance spectroscopy.

Hyaluronic acid (HA) is a high-value polysaccharide with many biomedical applications. Microbial production of HA is now replacing the traditional extraction method from rooster combs. Production of medical-grade HA with defined characteristics requires controlled process conditions because there are many fermentation process parameters that affect the microbial synthesis of HA. This necessitates the development of online tools for monitoring multiple analytes during microbial fermentation. Here, we describe the application of in situ transflectance spectroscopy for online quantification of seven major fermentation analytes, viz. biomass, glucose, lactate, formate, ethanol, acetate and HA in metabolically engineered Lactococcus lactis fermentations. The near-infrared spectral information acquired from synthetic mixtures and untransformed L. lactis fermentations were used to develop chemometric models. Based on principal component analysis and partial least squares regression methods, analyte-specific models were developed for quantification. These models were then independently validated for fermentation analytes from four different recombinant L. lactis strains. The chemometric model developed for HAbased on recombinant L. lactis fermentation data and pure HA standardscould accurately predict HA concentrations under homolactic conditions. The online estimation of HA was found to be poor under heterolactic conditions due to the overlapping absorbance of acetate produced in these cultures. Alternatively, an independent model based on yield correlation was successfully developed for indirect real-time quantification of HA.

Study for investigation of symptomatic improvement and safety of the ingestion of rooster comb degradation product containing low-molecular hyaluronic acid (INJUV) in individuals with knee and lower back pain; open-label trial with no control group

Study for investigation of symptomatic improvement and safety of the ingestion of rooster comb degradation product containing low-molecular hyaluronic acid (INJUV) in individuals with knee and lower back pain; open-label trial with no control group

Effectiveness of a low-fat yoghurt supplemented with rooster comb extract on muscle strength in adults with mild knee pain and mechanisms of action on muscle regeneration.

To determine the effects of the intake of low-fat yoghurt supplemented with rooster comb extract (RCE) on muscle strength. 148 subjects, with mild knee pain, participated in a randomized, placebo-controlled, double-blind, and parallel study. Muscle strength, knee effusion, and pain perception were measured. C2C12 myoblasts were used to elucidate the mechanisms of action involved. RCE improved total work and mean power in men, and also peak torque in extension by 10%. RCE reduced synovial effusion by 11.8% and pain perception by 24.6%. Both RCE and HA increased myoblast proliferation by 29%, while RCE reduced myoblast differentiation by 36.2%, suggesting a beneficial role of RCE in muscle regeneration. Low-fat yoghurt supplemented with RCE improved muscle strength. This effect is partially explained by muscle regeneration enhancement, reduced synovial effusion, and reduced pain perception, which could exert a beneficial clinical impact on men affected by mild knee pain.

Absence of differences among low, middle, and high molecular weight hyaluronan in activating murine immune cells in vitro

Hyaluronan (HA) effects on immune response are suggested to be dependent on HA molecular weight (MW), as low MW HA should activate immune cells in contrast to high MW HA. However, some current studies do not support this conception and emphasize the importance of the form of preparation of HA, particularly with respect to its purity and origin. We compared the activation of mouse immune cells by HA samples (100kDa, 500kDa, and 997kDa) prepared from HA originating from rooster comb, and HA samples (71kDa, 500kDa, and 1000kDa) prepared from pharmacological grade HA originating from Streptococcus equi. Interestingly, in contrast to established theory, only middle and high MW HA originating from rooster comb induced the production of tumor necrosis factor-α by macrophages and in whole blood. Further, all tested preparations of HA failed to induce the expression of inducible nitric oxide synthase, the production of nitric oxide, or the expression of cyclooxygenase 2 in macrophages and splenocytes. Importantly, all HA samples originating from rooster comb were found to be contaminated by endotoxin (up to 1.23EU/ml). Hence, low MW HA did not reveal itself to have significantly higher immunostimulatory activity compared to HA of higher MW.

Effect of ultrasonic degradation of hyaluronic acid extracted from rooster comb on antioxidant and antiglycation activities

Content: Recently, low-molecular-weight hyaluronic acid (LMWHA) has been reported to have novel features, such as free radical scavenging activities, antioxidant activities and dietary supplements. Objective: In this study, hyaluronic acid (HA) was extracted from rooster comb and LMWHA was obtained by ultrasonic degradation in order to assess their antioxidant and antiglycation activities. Materials and methods: Molecular weight (Mw) and the content of glucuronic acid (GlcA) were used as the index for comparison of the effect of ultrasonic treatment. The effects on the structure were determined by ultraviolet (UV) spectra and Fourier transform infrared spectra (FTIR). The antioxidant activity was determined by three analytical assays (DPPH, NO and TBARS), and the inhibitory effect against glycated-BSA was also assessed. Results: The GlcA content of HA and LMWHA was estimated at about 48.6% and 47.3%, respectively. The results demonstrate that ultrasonic irradiation decreases the Mw (1090–181 kDa) and intrinsic viscosity (1550–473 mL/g), which indicate the cleavage of the glycosidic bonds. The FTIR and UV spectra did not significantly change before and after degradation. The IC50 value of HA and LWMHA was 1.43, 0.76 and 0.36 mg/mL and 1.20, 0.89 and 0.17 mg/mL toward DPPH, NO and TBARS, respectively. Likewise LMWHA exhibited significant inhibitory effects on the AGEs formation than HA. Discussion and conclusion: The results demonstrated that the ultrasonic irradiation did not damage and change the chemical structure of HA after degradation; furthermore, decreasing Mw and viscosity of LMWHA after degradation may enhance the antioxidant and antiglycation activity.

Detection of hyaluronic acid in the aquatic waste and extraction by enzyme technology

Hyaluronic acid (HA) is a linear polysaccharide composed of a repeating disaccharide unit of β(1-4) glucuronic acid and β(1-3) N-acetylglucosamine. HA is present in tissues as cartilage, skin, rooster combs, umbilical cord, vitreous humour and sinovial fluid. In recent years, an increasing interest has been reported due to its numerous applications as cosmetic and pharmaceutical compound. HA levels from Carcharhinus sorrah, Dasyatis kuhlii and Pangasius bocourti lived in the sea area of Vietnam were the first time investigated. The extract of hyaluronic acid was dialyzed and digested with ENV enzyme. The optimal conditions of ENV protease for hydrolyzing cartilage and skin were determined as follows Carcharhinus sorrah: 1.25 IU enzyme/30gram of cartilage, 18 h, pH 6,0 at 60oC; Dasyatis kuhlii: 1.50 IU enzyme/30gram of cartilage, 30 h, pH 6,0 at 65oC and Pangasius bocourti: 1.50 IU enzyme /1.5gram of skin, 30 h, pH 6,0 at 65oC).

Geology of the Mid-Miocene Rooster Comb Caldera and Lake Owyhee Volcanic Field, eastern Oregon: Silicic volcanism associated with Grande Ronde flood basalt

The Lake Owyhee Volcanic Field (LOVF) of eastern Oregon consists of rhyolitic caldera centers and lava fields contemporaneous with and spatially related to Mid-Miocene Columbia River flood basalt volcanism. Previous studies delineated two calderas in the southeastern part of LOVF near Owyhee Reservoir, the result of eruptions of two ignimbrites, the Tuff of Leslie Gulch and the Tuff of Spring Creek. Our new interpretation is that these two map units are differentially altered parts of a single ignimbrite produced in a major phreatomagmatic eruption at ~15.8Ma. Areas previously mapped as Tuff of Spring Creek are locations where the ignimbrite contains abundant clinoptilolite±mordenite, which made it susceptible to erosion. The resistant intracaldera Tuff of Leslie Gulch has an alteration assemblage of albite±quartz, indicative of low-temperature hydrothermal alteration. Our new mapping of caldera lake sediments and pre- and post-caldera rhyolitic lavas and intrusions that are chemically similar to intracaldera Tuff of Leslie Gulch point to a single ~20×25km caldera, which we name the Rooster Comb Caldera. Erosion of the resurgently uplifted southern half of the caldera created dramatic exposures of intracaldera Tuff of Leslie Gulch cut by post-caldera rhyolite dikes and intrusions that are the deeper-level equivalents of lava domes and flows that erupted into the caldera lake preserved in exposures to the northeast.The Rooster Comb Caldera has features in common with more southerly Mid-Miocene calderas of the McDermitt Volcanic Field and High Rock Caldera Complex, including formation in a basinal setting shortly after flood basalt eruptions ceased in the region, and forming on eruption of peralkaline ignimbrite. The volcanism at Rooster Comb Caldera postdates the main activity at McDermitt and High Rock, but, like it, begins ~300ky after flood basalt volcanism begins in the area, and while flood basalts don't erupt through the silicic focus, are contemporaneous with the latest stages of eruptions nearby. High Rock and McDermitt rhyolites are associated with propagation of Steens Basalt dikes to the south, and LOVF rhyolites with later propagation of Grande Ronde Basalt dikes to the north and north-northwest.

Lack of in vitro antihyaluronidase activity of methanolic leaf extract of Indigofera tinctoria L and methanolic stem bark extract of Stereospermum suaveolens DC

Objective: To assess the antihyaluronidase activity of methanolic leaf extract of Indigofera tinctoria L (I. tinctoria) (family: Fabaceae/Leguminosae) and stem bark extract of Stereospermum suaveolens DC (S. suaveolens) (family: Bignoniaceae) in vitro with a view to develop an antiaging skin formulation. Materials and Methods: The antihyaluronidase activity of different concentrations (0.19 mg/mL, 0.38 mg/mL, 0.75 mg/mL, 1.5 mg/mL, and 3.0 mg/mL) of methanolic leaf extract of I. tinctoria, methanolic stem bark extract of S. suaveolens, and reference drug epigallocatechin gallate (EGCG) of different concentrations (12.5 μg/mL, 25 μg/mL, 50 μg/mL, 100 μg/mL, and 200 μg/mL) were determined spectrophotometrically using hyaluronic acid (from rooster combs) and bovine testicular hyaluronidase. Results: There is no in vitro antihyaluronidase activity in the methanolic extracts of I. tinctoria leaves and S. suaveolens stem bark even at high concentrations. On the contrary, EGCG, the reference agent, showed marked concentration-dependent (r2 = 0.92) antihyluronidase activity [in terms of percentage inhibition: half maximal inhibitory concentration (IC 50 ) 92.64 ± 0.64 μg/mL]. Conclusion: It is unlikely that skin antiaging effects of I. tinctoria leaves and S. suaveolens stem bark, as claimed in traditional and folk medicines in Sri Lanka, are mediated via antihyaluronidase activity.