Protein Content Of Extracts Research Articles

Enhancing zein functionality through sequential limited Alcalase hydrolysis and transglutaminase treatment: Structural changes and functional properties

This study investigated the effects of sequential enzymatic hydrolysis using Alcalase, followed by transglutaminase conjugation on the secondary and tertiary structures, hydrophobicity, free amine content, protein-protein interactions, and functional properties of zein. Fourier-transform infrared spectroscopy showed that the most significant secondary structural changes, characterized by a decrease in α-helix content and an increase in β-turns, occurred at a higher degree of hydrolysis. At a 2 % degree of hydrolysis, it revealed notable emulsifying activity (65.96 m2/g), while at 5 % hydrolysis, it achieved the highest solubility (75.06 %). Additionally, the zein hydrolysate with a 7 % hydrolysis degree, treated with transglutaminase, demonstrated improved H0 values (2992.33), enhanced foam capacity (65.95 %), and increased solubilized protein content in a dithiothreitol extract It can hydrolyze zein into numerous amphipathic polypeptide fragments, resulting in the generation of ant (31.35 %). Meanwhile, native zein treated with transglutaminase showed the highest water holding capacity (4.47 g/g). Overall, the combined enzymatic approach modified zein structure and properties, suggesting potential for improving functionality in plant-based food applications.

Impact of alkaline solution, carbohydrase and biomass:solvent on extraction efficiency, and protein and amino acid profiles of Palmaria palmata protein extracts

The impact of using different alkaline solutions, carbohydrases and changing the biomass to solvent volume ratio (biomass:solvent) on protein content, protein and biomass yield recovery in protein enriched extracts from Palmaria palmata (red seaweed) was studied. Protein extraction with alkaline solutions, i.e., NaOH, KOH, Ca(OH)2 and Na2CO3 (at 0.12 M) along with (and without) carbohydrases B, BD and L (containing xylanase, a mixture of xylanase and cellulase, and a combination of cellulase, xylanase and polygalacturonase activities, respectively) was assessed. Principal component analysis was used to assess correlations (if any) between the different alkaline solvents, carbohydrases and changing biomass:solvent on extraction efficiency. The highest protein content (56.64 ± 0.43 % (w/w)) was obtained using KOH assisted extraction with carbohydrase BD. The highest protein and biomass yield recoveries (70.65 ± 1.38 and 21.99 ± 0.43 % (w/w), respectively) were obtained on extraction using NaOH assisted with carbohydrases BD. Electrophoretic, amino acid profile and amino acid score analyses displayed differences depending on the alkaline solvent used for protein extraction. Appropriate selection of the alkaline solvent, carbohydrase and biomass:volume combination during protein extraction can contribute to the production of compositionally enhanced high-quality protein concentrates from P. palmata.

ELISA-4-amyloid: diagnostic accuracy of an ELISA panel for typing the four main types of systemic amyloidosis in subcutaneous abdominal fat tissue samples

Background Reliable typing of amyloid is essential. Amyloid extraction from tissue enables immunochemical typing of the precursor protein using an enzyme-linked immunosorbent assay (ELISA). Objective To assess the diagnostic accuracy of a panel of ELISAs for typing the four main types (AA, ATTR, AL-kappa and AL-lambda amyloid). Methods From 1996 to 2023 subcutaneous abdominal fat tissue aspirates were obtained from 1339 amyloidosis patients and 868 controls. Amyloid was visually graded 0–4+ in Congo red-stained smears. Amyloid extracted from tissue by Guanidine was typed using a panel comprising four ELISAs. Results All amyloid protein concentrations in extracts correlated with amyloid grade in smears. Typing sensitivity was low (23.3%) in samples with grade 1+/2+ amyloid. Overall typing sensitivity of the panel was 81.6% for all easily visible amyloid (grade 3+/4+): high for AA (98.8%) and ATTR (96.8%) and fair for AL-kappa (66.7%) and AL-lambda (75.9). Overall typing specificity was 98.0% and the overall positive predictive value was 98.0%. Conclusions We describe a highly specific ELISA panel for routine typing of the main amyloid types in fat tissue. Until more sensitive typing techniques will become generally available, typing easily visible amyloid in fat tissue using this ELISA panel is reliable, affordable and straightforward.

Supported ionic liquids to purify microbial L-Asparaginase

L-Asparaginase (ASNase) is a versatile enzyme that converts L-asparagine into ammonia and aspartic acid. This enzyme has applications in the food industry and health sector. However, high purity ASNase is required, resulting in high production costs. Therefore, in this work, two supported ionic liquids (SILs), specifically silica modified with dimethylbutylpropylammonium chloride ([Si][N3114]Cl) or triethylpropylammonium chloride ([Si][N3222]Cl), were investigated as alternative adsorption materials to purify ASNase. Different conditions were evaluated to improve enzyme purity, including total protein content in the cell extract, contact time, and SIL/cell extract ratio (w/v). Under optimal conditions using [Si][N3114]Cl, a maximum ASNase purification of 6.1-fold is achieved in a single step, resulting from the preferential attachment of other proteins on [Si][N3114]Cl SIL. According to the results, hydrophobic interactions rule the selective adsorption of protein impurities from the cell extract by the SIL, thereby increasing the ASNase purification levels. This approach offers a significant advantage, not requiring the desorption and elution of the target enzyme, while envisioning the application of SILs in a flow-through elution approach. The protonation state of protein surface was calculated by computational analysis, revealing that positively charged amino acids such as arginine and lysine block the effective binding of the enzyme to the SILs. Overall, if properly designed, SILs are promising alternative supports for the downstream processing of ASNase from cell extracts.

Proteins from shrews’ venom glands play a role in gland functioning and venom production

Venom production has evolved independently many times in the animal kingdom, although it is rare among mammals. Venomous shrews produce venom in their submandibular salivary glands and use it for food acquisition. Only a few toxins have been identified in shrew venoms thus far, and their modes of action require investigation. The biological and molecular processes relating to venom production and gland functioning also remain unknown. To address this gap, we investigated protein content in extracts from venom glands of two shrew species, Neomys fodiens and Sorex araneus, and interpreted their biological functions. Applying a proteomic approach coupled with Gene Ontology enrichment analysis, we identified 313 and 187 putative proteins in venom glands of N. fodiens and S. araneus, respectively. A search of the UniProt database revealed that most of the proteins found in both shrew species were involved in metabolic processes and stress response, while GO enrichment analysis revealed more stress-related proteins in the glands of S. araneus. Molecules that regulate molecule synthesis, cell cycles, and cell divisions are necessary to enable venom regeneration and ensure its effectiveness in predation and food hoarding. The presence of proteins involved in stress response may be the result of shrews’ high metabolic rate and the costs of venom replenishment. Some proteins are likely to promote toxin spreading during envenomation and, due to their proteolytic action, reinforce venom toxicity. Finally, finding numerous proteins involved in immune response suggests a potential role of shrew venom gland secretions in protection against pathogens. These findings open up new perspectives for studying biological functions of molecules from shrew venom glands and extend our knowledge on the functioning of eulipotyphlan venom systems. Because the majority of existing and putative venomous mammals use oral venom systems to inject venom into target species, the methods presented here provide a promising avenue for confirming or discovering new taxa of venomous mammals.

Antiproliferative activity <i>in vitro</i> of <i>Trichinella spiralis</i> larval extracts depending on the infection stage following experimental infection of rats

Relevance.The aim is evaluation in vitro antiproliferative activity of T. spiralis larvae extracts obtained from experimentally infected rats, depending on the duration of infection.Methods. The experiment was carried out on outbred male rats infected with T. spiralis larvae at the rate of 10 larvae per 1 g of body weight. The antiproliferative activity of larval extracts obtained 40 days, 3 and 12 months after infection was determined on tumor cells of the A549 line (human lung carcinoma) using an MTT test during cultivation for 24, 48 and 72 hours in concentrations from 32 to 1000 micrograms/ml. Antiproliferative activity was expressed as % inhibition and IC50.Results. There was direct dependence of Trichinella extract antiproliferative effects on time of cultivation with A549 cells and protein concentration in extracts. At all stages of invasion, the antiproliferative activity of the extract increased from 24 to 72 hours. at a protein concentration of 250 to 1000 mcg/ml. The extract antiproliferative activity expressed in tumor cell % inhibition and IC50 increased from day 40 to month 12. The most intense dynamics was observed between day 40 and month 3. The possible reason of enhancement in extract antiproliferative effect at months 3 and 12 is alteration in quantitative and qualitative composition of proteins comprised the extracts.

Pengaruh suhu berbeda terhadap aktifitas enzim kolagenase dari usus ikan cunang (Congresox talabon)

Collagenase enzyme is a type of protease enzyme. The activity of an enzyme is affected by substrate, temperature, inhibitor, and pH. One of the sources of enzymes is the internal organs of fish, namely the gut. This study aims to determine the activity of the collagenase enzyme at different temperatures. The experimental design used was randomized and full non-factorial. The treatments used were different incubation temperatures (50, 55, and 60oC) with three replications. Parameter analysis comprised fish proportion, enzyme activity, and dissolved protein. The results showed that the parts of the yellow pike conger fish were meat (64.25%), bones (19.98%), internal organs (including intestines) (4.50%), and skin (6.30%). Collagenase enzyme activity at temperatures 50, 55, and 60oC was 1.154, 1.702, and 0.667 U/mL, respectively. The dissolved proteins content was 12.89 mg/mL. The optimum temperature of the collagenase enzyme from the fish gut was 55oC with an enzyme activity of 1,702 U/mL. The dissolved protein content of the gut yellow pike conger extract was 12.89 mg/mL Purification of the collagenase enzyme from of the gut yellow pike conger was needed to obtain a higher enzyme activity value.

Antioxidant and Antimicrobial Potential of Peptide from Rabbit Meat Hydrolysate Prepared by Trypsin and Zingibain.

<b>Background and Objective:</b> Rabbit meat is a livestock product potentially viable as a protein source to obtain peptides. Antioxidant and antimicrobial peptides are ingredients extracted from various foods through enzymatic hydrolysis, chemical hydrolysis and fermentation to produce health-promoting foods. This research aims to investigate the potential of rabbit meat as a source of antioxidant and antimicrobial peptides through hydrolysis using trypsin and zingibain enzymes. <b>Materials and Methods:</b> This research conducted an explorative-descriptive approach, focusing on antioxidant and antimicrobial activity. Rabbit meat was extracted using trypsin, zingibain and a combination of trypsin and crude extract zingibain. The hydrolyzed rabbit meat extract was tested at intervals of 0, 2, 6, 16, 24, 40 and 48 hrs to determine the degree of hydrolysis and the profile of hydrolyzed proteins with electrophoresis SDS PAGE. The antioxidant activity was tested using the DPPH method and the antimicrobial activity using agar well diffusion method. <b>Results:</b> The degree of hydrolysis increased with the hydrolysis time. The highest protein content of rabbit meat extract hydrolyzed with trypsin was 287.65 mg/mL, observed during 12 hrs hydrolysis. The optimum conditions for the hydrolysis of rabbit meat protein were obtained at 24 hrs, with an IC<sub>50</sub> value of 52.45% hydrolyzed by trypsin. As per antimicrobial activities, <i>Escherichia coli</i> and <i>Salmonella</i> sp. were more effective in inhibiting rabbit meat hydrolysates compared to <i>Pseudomonas aeruginosa</i> and <i>Staphylococcus aureus</i>. The inhibition of all pathogen increased until 12 hrs hydrolysis but decreased in 24 hrs hydrolysis. <b>Conclusion:</b> The combination zingibain enzyme and trypsin is feasible for hydrolyzing rabbit meat and the optimum hydrolysis time was 24 hrs with IC<sub>50</sub> 52.45 ppm, although accompanied by reduction in antibacterial activities.

Characterization of Pollen Grain and Pollen Extract from Common Allergenic Plants in the Philippines

Allergic rhinitis and asthma are prevalent in tropical countries like the Philippines. Pollen grains from anemophilous plant species may trigger these respiratory allergies. Pollen extracts from common allergenic plants are currently produced locally in the Philippines and are utilized for the diagnosis and immunotherapy of patients with respiratory allergies. Further improvement of their quality is necessary to avoid false negative or false positive diagnoses, as well as to increase the effectiveness of immunotherapy and avoid adverse reactions. This study aimed to characterize the morphology of the collected pollen grains and assess the pollen purity, protein concentration, and electrophoretic profile of the allergenic pollen extracts. Pollen identification and percent purity of the pollen collections were assessed via microscopy and hemocytometer. Lowry assay was performed to identify the total protein concentration of the pollen extracts before subjecting them to SDS-PAGE and silver staining. The pollen samples from varying plant families have distinct features that will aid in their proper identification. Collected aggregates of pollen samples with pollen purity of < 90%, like T. procumbens, A. saman, L. leucocephala, and M. indica, manifest darker colors compared to their actual color. On the other hand, the color of collected pollen samples with pollen purity ≥ 90% are commonly yellow or off-white, which is their natural color. The total protein concentration of the crude pollen extracts ranges from 2.87–9.63 mg/mL. The pollen extracts revealed protein bands at molecular weights ranging from 10–250 kD on SDS-PAGE. Proper characterization of a particular pollen may help in assessing the purity of the collected pollen. Higher pollen purity may contribute to the assurance that the positive diagnosis was due to the target pollen of a plant and not due to contaminations from other components. The result of this study may be considered for the improvement of the crude pollen allergen extract production for clinical use. It is also recommended to assess the effect of the environmental condition where the pollen was collected in terms of the actual proteins present after allergen extraction.

Optimizing the Comet Assay-Based In Vitro DNA Repair Assay for Placental Tissue: A Pilot Study with Pre-Eclamptic Patients.

The comet assay-based in vitro DNA repair assay has become a common tool for quantifying base excision repair (BER) activity in human lymphocytes or cultured cells. Here, we optimized the protocol for studying BER in human placental tissue because the placenta is a non-invasive tissue for biomonitoring of early-life exposures, and it can be used to investigate molecular mechanisms associated with prenatal disorders. The optimal protein concentration of placental protein extracts for optimal damage recognition and incision was 2 mg protein/mL. The addition of aphidicolin did not lead to reduced non-specific incisions and was, therefore, not included in the optimized protocol. The interval between sample collection and analysis did not affect BER activity up to 70 min. Finally, this optimized protocol was tested on pre-eclamptic (PE) placental tissues (n = 11) and significantly lower BER activity in PE placentas compared to controls (n = 9) was observed. This was paralleled by a significant reduction in the expression of BER-related genes and increased DNA oxidation in PE placentas. Our study indicates that BER activity can be determined in placentas, and lower activity is present in PE compared with healthy. These findings should be followed up in prospective clinical investigations to examine BER's role in the advancement of PE.

Fermentation profile, chemical composition, and aerobic stability of cassava shoots silages with cactus pear

The aim was to evaluate the effect of the cactus pear levels (0, 15, 30, and 45%) in cassava shoot silages on the fermentation profile, chemical composition, and aerobic stability. Four treatments were tested with five repetitions, totaling 20 silos. The inclusion of cactus pear in cassava shoots silages promoted an increase in gas losses (P < 0.001), buffering capacity (P < 0.001), pH (P = 0.033), mineral matter (P < 0.001), total carbohydrates (P < 0.001), non-fiber carbohydrates (P < 0.001) and a decreasing in dry matter (P < 0.001), organic matter (P < 0.001), ether extract (P = 0.002), and crude protein (P < 0.001) content. A quadratic effect was observed for effluent losses (P < 0.001), with greater effluent losses (94.78 kg/t natural matter) obtained with the inclusion of 30% cactus pear in cassava shoots silages. Cactus pear inclusion in cassava shoots silage promoted a reduction in the pH during oxygen exposure (P = 0.008). Including cactus pear in up to 45% of cassava silage, shoots cause changes in the fermentation and nutritional characteristics. However, values found in the silages are by good quality standards.

Optimizing protein extraction from chickpea flour: buffer and temperature effects

Legumes constitute a significant protein source in the human diet. Among this category, the chickpea (Cicer arietinum L.) is recognized as an alternative crop due to its efficient water usage. This efficiency has generated a growing interest in its improvement, commercialization and generation of value-added products, particularly within the domain of functional foods. The literature contains a multitude of techniques for evaluating protein content in legumes, complicating the comparison of results among different research groups. The aim of this study was to assess the effect of four distinct extraction buffers on protein concentration in extracts of chickpea flour from two varieties, Costa 2004 and Blanoro, testing the effect of the extraction temperature at 25 and 37 °C. Additionally, the integrity of the samples was evaluated by analyzing the protein profile using SDS-PAGE. The results obtained showed a significant difference in protein concentration due to the choice of extraction buffer and extraction temperature. We propose the implementation of Tris buffer 500 mM pH 6.8 at 25°C to standardize the process of characterizing protein extracts from chickpea flour for comparative purposes. This standardized protocol serves as a valuable tool for agronomic characterization and offers a comprehensive framework for future research in this field.

The cross-linking and protective effect of artemisinin and its derivatives on collagen fibers of demineralized dentin surface

ObjectiveTo investigate the cross-linking and protective effect of artemisinin (ART), dihydroartemisinin (DHA), and artesunate (AST) on collagen fibers of demineralized dentin surface. MethodsMolecular docking was used to predict potential interactions of ART, DHA, and AST with dentin type I collagen. Human third molars without caries were completely demineralized and treated with different solutions for 1 min. The molecular interactions and cross-linking degree of ART and its derivatives with dentin collagen were evaluated by FTIR spectroscopy, total extractable protein content, and a ninhydrin assay. Scanning electron microscopy, hydroxyproline release, and ultimate microtensile strength tests (μUTS) were employed to confirm the mechanical properties and anti-collagenase degradation properties of dentin collagen fibers. ResultsART, DHA, and AST combined with dentin type I collagen mainly through hydrogen bonding and hydrophobic interactions, and the cross-linking reaction sites were mainly C=O and CN functional groups. Compared to the control group, ART and its derivatives significantly increased the degree of cross-linking. Additionally, significant increases were observed in resistance to enzymatic digestion and mechanical properties of the artemisinin and its derivatives group. ConclusionART, DHA, and AST could cross-link with demineralized dentin collagen, through improving the mechanical properties and anti-collagenase degradation properties. Clinical significanceThe study endorses the potential use of ART and its derivatives as a prospective collagen cross-linking agent for degradation-resistant and long-period dentin bonding in composite resin restorations.

Evaluation of the contents and ratios of five fractionated proteins to elucidate the protein composition in soybean seeds

AbstractSelecting suitable soybean cultivars is important for food processing and exploring their endowment with desirable physiological functions. We applied the fractionation method previously established to compare soy protein composition among cultivars to promote such a selection. More than 95% of the proteins in soybean cotyledons were extracted from 13 soybean cultivars using a high‐concentration salt solution. The extracted proteins were fractionated into five fractions, namely oil body‐associated protein (OBAP), polar lipid‐associated protein (PLAP), globulins (11S and 7S), and whey by centrifugation after tuning the solubility behavior of the proteins with various solutions. Protein species in each fraction were analyzed by SDS‐PAGE. Protein content in the total extract and five fractions was quantified to characterize the protein composition of soybean cultivars. The correlation between the protein content of each fraction and the total protein in cotyledon was investigated. A strong positive correlation was found only for the 11S fraction (r = 0.82), followed by a positive correlation in the 7S fraction (r = 0.65). Thus, we surmised that the increased protein content in soybean was due to increased globulin content. Furthermore, the calculation of the average ratio of protein content in each fraction indicated the globulin fraction (7S and 11S) to be 52%, the lipophilic protein fraction (OBAP and PLAP) to be 33%, and the whey fraction to be 13%. The preparation method employed in this study is a promising tool for efficiently comparing the protein composition of soybean cultivars to evaluate the potential use of cultivars for food production.

Investigation of cryoprotectants-treated surimi protein deterioration during chilled and frozen storage: Functional properties and kinetic modeling.

The relative cryoprotective effects of flaxseed protein hydrolysate and pectin in comparison with conventional cryoprotectant (sucrose + sorbitol + sodium tripolyphosphates) on stabilization of proteins in surimi of Capoor (Cyprinus carpio) were investigated during freezing (-20°C for 4 months) and chilling storage (4°C for 10 days). Although pectin caused to improve water-holding capacity (27.8%; 4°C and 21.5%; -20°C) on account of highly more inhibitory impact on the ice crystals growth, the protein denaturation may have occurred. It can be related to higher reduction in the amount of salt extractable protein (%) and the immeasurable value of thiol group in surimi formulation containing pectin compared with other cryoprotectants. The results of modeling surimi samples showed that salt extractable protein and sulfhydryl content were in good agreement with the first-order reaction model at -20°C and second-order kinetic model at 4°C. In comparison with other samples, samples treated with flaxseed protein showed the lowest reaction rate constant during chilled and frozen storage. The results confirmed that flaxseed protein with no sweetness and considerable caloric value had a cryoprotective effect similar to sucrose + sorbitol + polyphosphate and even better.

UJI AKTIVITAS ENZIM SUPEROKSIDA DISMUTASE (SOD) EKSTRAK DAUN SALAM (Syzygium polyanthum) DENGAN METODE WATER SOLUBLE TETRAZOLIUM SALT-1 (WST-1)

Free radicals can cause various kinds of degenerative diseases and skin aging. Free radicals can be neutralized with an enzymatic antioxidant, namely superoxide dismutase (SOD). One of the plants that has high SOD activity is salam (Syzygium polyanthum). The aim of this study was to determine the activity of the crude SOD extract of bay leaves precipitated with ammonium sulfate salt at various concentrations of 25, 50, 75%. The study began with plant determination, enzyme extraction, precipitation of enzyme extracts with ammonium sulfate concentrations of 25, 50, 75%, measurement of total protein content and SOD activity test using the WST-1 method to calculate the percent inhibition. The results were statistically analyzed using one way ANOVA. The results showed that the total protein content of the crude extract of SOD and the results of precipitation with ammonium sulfate 25, 50, and 75% respectively were 9.38; 6.36; 8.50; and 11.6 mg/mL. Meanwhile, the percentage inhibition values were 73.13; 34.33; 65.67; and 80.59%. The highest SOD activity was obtained in the crude extract of the enzyme precipitated with 75% ammonium sulfate salt, which was 80.59%.

KARATERISTIK ENKAPSULASI LIPOSOM EKSTRAK SUPEROKSIDA DISMUTASE (SOD) Bacillus altitudinis

Photoaging causes the dermis and collagen fibers to decrease which results in skin aging. Genus Bacillus has SOD activity as an anti-photoaging agent that can neutralize ROS (Reactive Oxygen Species) due to exposure to UV-A rays. This study aimed to examine the activity of crude extract and the encapsulation characteristics of liposomes from B. altitudinis. The study began with the isolation of the SOD enzyme, determination of the total protein content of the crude extract of the SOD enzyme and liposome encapsulation, then characterization liposome tes including particle size analysis, sorption efficiency and zeta potential. The results of the crude extract activity test were 85.09% ± 1.24%. The characteristic test was produced with a size of 112.96 ± 17.3 ; the adsorption efficiency was 81.1% and the zeta potential was 7.0 ±5.1 mV. The results showed that the SOD B. altitudinis liposomes were in the nanoparticle range.

Anti-microbial activity of seed oils & protein estimation of seed-content in Clitoria ternetea, Lagenaria siciraria & Ziziphus mauritiana

Asian plants of flower, vegetable and fruit named Clitoria ternetea, Lagenaria siciraria and Ziziphus mauritiana respectively are known for their extensive uses in traditional medicine as well as traditional treatment of various diseases and infections. Several studies have already been done so far on these plants and its various parts but the studies on their seeds as well as their seed oilwas lacking. In this work, we determined the protein content of these seeds as well as we extracted the seed oils and determined their anti-microbial activity independently along with the anti-microbial activity by mixing all these oils together in equal ratios. Both, protein determination and anti-microbial activity determination showed considerable and potential results. The Lagenaria siciraria seeds (in ethanol extract) showed highest protein content amongst all i.e., 4.44 (μg/mL-1) whereas Clitoria ternetea seeds displayed highest protein content (in water extract) amongst all i.e., 5.07 (μg/mL-1). The anti-microbial activity of the mixed seed oil showed better and great results as compared to individual anti-microbial activity of seed oils.

The fatty acid composition, phytochemicals and antioxidant potential of wild edible Smilax excelsa L. shoots.

The present study examines the fatty acid composition, phytochemicals content and antioxidant potential of Smilax excelsa L. shoots, which grows spontaneously and is consumed as food in rural areas of Düzce (Türkiye) province, was investigated. DPPH, ABTS and OH radical scavenging tests were utilized to put forth the antiradical properties of the extracts of this plant. In addition, the metal chelating potential of this plant was also evaluated. The higher the inhibition % value calculated in these tests, the higher the antioxidant activity was considered and the results were evaluated. The average ABTS radical cleaning test results of Smilax excelsa extracts prepared in different concentrations of methanol, ethanol and pure water were found to be 98.14%, 98.16%, 90.20%, respectively. The average DPPH radical cleaning test results of the extracts of this plant prepared in different concentrations of methanol, ethanol and pure water were determined as 87.48%, 76%, 46.53%, respectively. The OH radical cleaning test results of methanol, ethanol and pure water extracts of the Smilax excelsa were determined as 54.79%, 72.54%, none, respectively. In addition, the metal chelation test results of methanol, ethanol and pure water extracts of this plant were determined as 87.26%, 89.36%, 53.70%. The highest protein (85.91 mg BSA/g), proanthocyanidin (55.39 mg CE/g) and phenolic (4957.57 μg GAE/g) content of Smilax excelsa pure water extract was determined. It has been determined that gallic acid (117.33 μg/g), vanillic acid (33.89 μg/g), caffeic acid (4.55 μg/g), ferulic acid (93.78 μg/g), rosmarinic acid (0.33 μg/g) and hydrocynamic acid (0.33 μg/g) are found in different proportions in the Smilax excelsa. It has been stated that Smilax excelsa is an important source of palmitic acid (20.52%), stearic acid (4.95%), oleic acid (4.74%), linoleic acid (20.99%), γ-linolenic acid (2.26%), alpha-linolenic acid (34.29%) and docosahexaenoic acid (2.79%). It has been found that this plant has a low content of fat-soluble vitamins and phytosterols, with the exception of β-sitosterol (6.43μg/g).

Evaluation and Comparison of the Effects of Mature Silkworm (Bombyx mori) and Silkworm Pupae Extracts on Schwann Cell Proliferation and Axon Growth: An In Vitro Study.

Silkworm products were first used by physicians more than 8500 years ago, in the early Neolithic period. In Persian medicine, silkworm extract has several uses for treating and preventing neurological, cardiac, and liver diseases. Mature silkworms (Bombyx mori) and their pupae contain a variety of growth factors and proteins that can be used in many repair processes, including nerve regeneration. The study aimed to evaluate the effects of mature silkworm (Bombyx mori), and silkworm pupae extract on Schwann cell proliferation and axon growth. Silkworm (Bombyx mori) and silkworm pupae extracts were prepared. Then, the concentration and type of amino acids and proteins in the extracts were evaluated by Bradford assay, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and liquid chromatograph-mass spectrometer (LC-MS/MS). Also, the regenerative potential of extracts for improving Schwann cell proliferation and axon growth was examined by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay, electron microscopy, and NeuroFilament-200 (NF-200) immunostaining. According to the results of the Bradford test, the total protein content of pupae extract was almost twice that of mature worm extract. Also, SDS-PAGE analysis revealed numerous proteins and growth factors, such as bombyrin and laminin, in extracts that are involved in the repair of the nervous system. In accordance with Bradford's results, the evaluation of extracts using LC-MS/MS revealed that the number of amino acids in pupae extract was higher than in mature silkworm extract. It was found that the proliferation of Schwann cells at a concentration of 0.25 mg/mL in both extracts was higher than the concentrations of 0.01 and 0.05 mg/mL. When using both extracts on dorsal root ganglion (DRGs), an increase in length and number was observed in axons. The findings of this study demonstrated that extracts obtained from silkworms, especially pupae, can play an effective role in Schwann cell proliferation and axonal growth, which can be strong evidence for nerve regeneration, and, consequently, repairing peripheral nerve damage.