Objective A subcutaneous transplantation tumor model of human HT-29 cells in nude mice was established to evaluate anticarcinogenic activities, and the apoptosis-regulated mechanism effect of aqueous extract of fermen...Objective A subcutaneous transplantation tumor model of human HT-29 cells in nude mice was established to evaluate anticarcinogenic activities, and the apoptosis-regulated mechanism effect of aqueous extract of fermented wheat germ with Lactobacillus plantarum dy-1 (LFWGE). Methods The HT-29 cells were transplanted via subcutaneous injection of 1×10^7cells into the right flank of each nude mouse. Then, nude mice were treated for 30 d with LFWGE (high-dose 2 g/kg/d; low-dose 1 g/kg/d) and for 7 d with 5-fluorouracil (5-FU, 25 mg/kg/d) by gavage and intraperitoneal injection, respectively. An inhibition of tumor growth was observed. Results Tumor volume and weights decreased significantly in both groups of nude mice treated with LFWGE. In addition, the cell apoptosis rate of the LFWGE group (2 g/kg/d, 60.2%+4.4%; 1 g/kg/d, 58.6%+6.9%) was significantly higher than that of the control group (11.5%+1.6%) and 5-FU group (32.1%+3.5%) as measured by the TUNEL assay. Moreover, the real-time fluorescent quantitative PCR and Western blot method further confirmed these enhancing apoptosis and growth inhibition effects. The involvement of LFWGE in inducing apoptosis was confirmed by the expression of Bax, Bcl-2, Caspase-3, and CyclinD1. Conclusion The results showed that LFWGE could induce subcutaneous transplantation tumor apoptosis in nude mice and could be as a natural nutrient supplements or chemopreventive agent in the treatment of human colon cancer.展开更多
An optical biosensor with a stirred cuvette has been used to monitor the interaction of immobilized wheat germ agglutinin (WGA) with two water-soluble cationic porphyrins, The association constants (Ka) of the fre...An optical biosensor with a stirred cuvette has been used to monitor the interaction of immobilized wheat germ agglutinin (WGA) with two water-soluble cationic porphyrins, The association constants (Ka) of the free base porphyrin and its Zn(Ⅱ) complex form were 2. 66 and 27.31 × 10^5 l,/mol at 20 ℃ respectively. The interactions of the free base porphyrin were further investigated at temperatures between 15 ℃ and 37 ℃, The thermodynamics parameters, changes in free energy, enthalpy and entropy, were -31.23, 22.92, 54.15 ld/mol respectively. The heat capacity change was -355.53 J · mol^-1·K^-1 The binding was driven by entropic contribution, and showed strong enthalpy-entropy compensation. It was governed primarily by hydrophobic forces.展开更多
[Objectives]This study was conducted to develop a health-care product with smooth taste and high nutrition.[Methods]With natural wheat germ powder as the main raw material and maltodextrin,white sugar,compound stabili...[Objectives]This study was conducted to develop a health-care product with smooth taste and high nutrition.[Methods]With natural wheat germ powder as the main raw material and maltodextrin,white sugar,compound stabilizer and other materials as auxiliary materials,the formula of wheat germ solid beverage was optimized using sensory evaluation value as an index,through the mixture experiment method and fuzzy mathematical analysis.[Results]The optimal formula of wheat germ solid beverage was:sugar 11.51%,maltodextrin 37.89%,wheat germ powder 50%,compound stabilizer 0.6%(arabic gum 0.3%,xanthan gum 0.3%),soya bean lecithin 0.2%,ethyl maltol 0.002%,and vanillin 0.003%.[Conclusions]This study provides a theoretical basis for expanding the market-oriented production and application of wheat germ.展开更多
Several studies are conducted to improve nutritional enrichment of ice cream. The aim of the present study was to produce healthy ice cream by partial replacement of fat with whey protein (WP), dietary fibers (oat (OT...Several studies are conducted to improve nutritional enrichment of ice cream. The aim of the present study was to produce healthy ice cream by partial replacement of fat with whey protein (WP), dietary fibers (oat (OT) and wheat germs (WG)) and modified starch (MS). Reduced fat ice cream (8% fat) was prepared as control. Healthy reduced fat ice cream was prepared by replacing 1% and 2% of fat with WP, OT, WG and MS. The fat replacement with OT and WG increased the fiber content of ice cream. In addition, ice cream from all treatments characterized by a significant great free radical scavenging activity when compared with control, especially when fat was replaced with WG, WP and OT, respectively. The sensory evaluation of ice cream revealed an enhancement of flavor when fat was replaced with 1% WP and OT and 2% WP (P < 0.05 vs. control). The modified starch treatments (1% and 2%) showed the highest viscosity followed by OT (2%) treatment (P < 0.05). In conclusion, these results showed that dietary fibers (OT & WG), WP and MS could be used to replace fat in ice cream and to give a healthy reduced fat ice cream with high fiber content and with high antioxidant activity.展开更多
Wheat germ protein hydrolysates were prepared by protease hydrolysis, ultrafiltration and dynamical adsorption of resin. The total amount of amino acids in 100 g wheat germ protein hydrolysates is 93.95 g. Wheat germ ...Wheat germ protein hydrolysates were prepared by protease hydrolysis, ultrafiltration and dynamical adsorption of resin. The total amount of amino acids in 100 g wheat germ protein hydrolysates is 93.95 g. Wheat germ protein hydrolysates are primarily composed of 4 fractions: 17.78% in the relative molecular mass range of 115631512, 17.50% in 1512842, 27.38% in 842372 and 30.65% in 37276, respectively. The antioxidant properties of wheat germ protein hydrolysates were evaluated by using different antioxidant tests in vitro. 1.20 g/L wheat germ protein hydrolysates exhibit 78.75% inhibition of peroxidation in linolei acid system; and 1.6g/L wheat germ protein hydrolysates show 81.11% scavenging effect on the 1,1-diphenyl-2-picrylhrazyl radical. The reducing power of 2.50 g/L wheat germ protein hydrolysates is 0.84. Furthermore, the scavenging activity of 0.60 g/L wheat germ protein hydrolysates against superoxide radical is 75.40%; 0.50 g/L wheat germ protein hydrolysates exhibit 63.35% chelating effect on ferrous ion. These antioxidant activities of wheat germ protein hydrolsates increase with the increase of its concentration. Experimental results suggest that wheat germ protein hydrolysate is a suitable natural antioxidant rich in nutrition and nontoxic.展开更多
Wheat germ is a by-product derived from the wheat milling industry. Defatted wheat germ is the main by-product of the wheat germ in the oil extraction process. This study aims at development of efficient and low cost ...Wheat germ is a by-product derived from the wheat milling industry. Defatted wheat germ is the main by-product of the wheat germ in the oil extraction process. This study aims at development of efficient and low cost processing methods to transform these residues in added value co-product. In this study, wheat germ was analysed for its proximate composition, fatty acid composition, physical and chemical characteristics of wheat germ oil. The basic chemical composition analyses revealed high values of dry matter (87.37 g/100g FW), significant amounts of total protein and fat (27.69 and 8.99 g/100g FW, respectively) content and low ash content (3.08 g/100g FW). The quality of the extracted oils was assessed in terms of acid value, iodine value, saponification value, peroxide value, refractive index, and unsaponifiable matter. The fatty acid profile was found to be made up of linoleic followed by palmitic and oleic as the major fatty acids. Antioxidant properties and in vitro antibacterial activity of defatted wheat germ (DWG) extract were also determined. DWG, as a source of natural antioxidants and antibacterial, could be used to formulate nutraceuticals with potential applications to reduce the level of oxidative stress. The antioxidant potency of the DWG extracts could be the basis for its health promoting potential. The results showed that these by-products could be used as a source of bioactive compounds beneficial for health.展开更多
This work aimed at determining the maximum acceptable level of wheat germ that can be added to one of homemade biscuits in Jordan, studying some chemical and sensory properties of this product, moreover, studying the ...This work aimed at determining the maximum acceptable level of wheat germ that can be added to one of homemade biscuits in Jordan, studying some chemical and sensory properties of this product, moreover, studying the effect of storage time after two months at room temperature on the sensory parameters of this product. The results showed that the addition of wheat germ in biscuit making at level of 20% (w/w) enhanced the nutritional value by increasing protein and total mineral content, as well as decreasing fat, total carbohydrates and total calories. The acceptability of sensory characteristics was improved by this fortification immediately after preparation and during storage.展开更多
We describe a two-step method that uses wheat germ agglutinin immobilized on Sepharose gel followed by immunoaffinity chromatography (IAC) to extract recombinant human erythropoietin and Darbepoetin from equine plasma...We describe a two-step method that uses wheat germ agglutinin immobilized on Sepharose gel followed by immunoaffinity chromatography (IAC) to extract recombinant human erythropoietin and Darbepoetin from equine plasma. Lectin affinity chromatography was shown to be an effective approach for isolating these epoetins from plasma and in combination with IAC;this method gave superior recovery when compared to the use of the latter technique alone. Moreover, due to the ease with which it can be scaled up, it is particularly well suited for pre-concentrating larger volumes of samples prior to IAC and this provides a facile way of improving the overall sensitivity with which these foreign proteins can be detected in equine plasma.展开更多
为深入探究小麦胚芽中的脂质成分,该研究采用不同溶剂分别提取小麦胚芽中的磷脂和中性脂质,利用核磁共振技术(nuclear magnetic resonance,NMR)对小麦胚芽中的脂质成分进行分析,结果表明:^(31)P NMR在小麦胚芽中检测到6种磷脂和相对应的...为深入探究小麦胚芽中的脂质成分,该研究采用不同溶剂分别提取小麦胚芽中的磷脂和中性脂质,利用核磁共振技术(nuclear magnetic resonance,NMR)对小麦胚芽中的脂质成分进行分析,结果表明:^(31)P NMR在小麦胚芽中检测到6种磷脂和相对应的5种溶血性磷脂,以及非脂质含磷化合物甘油磷脂酰胆碱(GPC);磷脂中含量最高的是磷脂酰胆碱(PC),其摩尔浓度为0.42μmol/g,摩尔分数为28.50%,质量浓度为0.31 mg/g,质量分数为30.20%;^(1)H NMR测定小麦胚芽中的多种中性脂质组成及含量,包括甘油三酯(TG)、甘油二酯(DG)、甘油单酯(MG)和游离脂肪酸(FA),其中含量最高的甘油酯是TG,占比为77.25%,含量最低的是2-甘油单酯(2-MG),占比为0.03%;小麦胚芽的甘油三酯和磷脂中检测出亚油酸(L)、油酸(O)和亚麻酸(Ln)等6种不饱和脂肪酸,其中亚油酸的含量最高,在甘油三酯中占比56.26%,在磷脂中占比45.37%。溶血性磷脂和GPC是磷脂的水解产物,DG、MG和FA是甘油三酯的水解产物,这些物质可以反映样品中脂质的水解程度。研究结果表明,利用NMR不仅能够对小麦胚芽脂质的组分进行定性定量分析,而且可以监测小麦胚芽脂质水解程度的变化。NMR技术在小麦胚芽相关产品的脂质分析研究具有重要作用。展开更多
Background Pink bollworm,Pectinophora gossypiella(Saunders)(Lepidoptera:Gelechiidae)has become a poten-tial pest of cotton by causing substantial yield losses around the world including Pakistan.Keeping in view the fa...Background Pink bollworm,Pectinophora gossypiella(Saunders)(Lepidoptera:Gelechiidae)has become a poten-tial pest of cotton by causing substantial yield losses around the world including Pakistan.Keeping in view the facts like limited research investigations,unavailability,and high cost of artificial diet’s constituents and their premixes,the present research investigations on the dietary aspect of P.gossypiella were conducted.The larvae of P.gossypiella were reared on different diets that were prepared using indigenous elements.The standard/laboratory diet com-prised of wheat germ meal 34.5 g,casein 30.0 g,agar–agar 20.0 g,sucrose 10.0 g,brewer’s yeast 5.0 g,α-cellulose 1.0 g,potassium-sorbate1.5 g,niplagin 0.5 g,decavitamin 0.01 g,choline-chloride 0.06 g,maize-oil 3.30 g,honey 2.0 g,and water 730.0 mL.Alternatives to cotton bolls and wheat germ meal were okra seed sprouts,okra fruit,cottonseed meal,and okra seed meals,which were included in the study to introduce an efficient and economic mass-rearing system.Results The larval development completed in 19.68d±0.05 d with a weight of 20.18mg±0.20 mg at the fourth instar fed on the cottonseed meal-based diet instead of wheat germ meal based diet.On the same diet,84.00%±4.00%,17.24 mg±0.03 mg,and 7.76d±0.06 d were recorded as pupae formation,pupal weight,and pupal duration,respectively.Adult emergence,76.00%±1.00%was recorded from pupae collected from larvae raised on cottonseed meal-based diet.These male and female moths lived for 40.25d±0.10 d,and 44.34d±0.11 d,respectively.Females deposited 21.28±0.04 eggs per day with the viability of 65.78%±0.14%.The larval mortal-ity at the fourth instar was 37.20%±1.36%and malformed pupation of 12.00%±1.41%was recorded.Replacement of wheat germ meal with that of local meals(cottonseed and okra seed)in the standard laboratory diet has saved 463.80 to 467.10 PKR with 1.62 to 1.63 cost economic returns,respectively.Conclusion This research is of novel nature as it provides a concise and workable system for the economic and suc-cessful rearing of P.gossypiella under laboratory conditions.展开更多
In order to refine the products of wheat germ agglutinin(WGA),several ion exchangers,such as D261,732,DEAE-cellulose-32 and CM-cellulose-32, have been used to removed proteins and pigments with different colors,such a...In order to refine the products of wheat germ agglutinin(WGA),several ion exchangers,such as D261,732,DEAE-cellulose-32 and CM-cellulose-32, have been used to removed proteins and pigments with different colors,such as brown,red,yellow,green and black,in the extracts of wheat germ.The WGA obtained from this procedure has higher hemagglutination activity than that available from sigma Co. The minimum hemagglutination dose of the purified WGA for an equal volume of a 2% type A red blood cells is 4μg/ml.WGA is a mixture of isolectins with different isoelectric points,5.9,6.2 and 6.8.Their molecular weight identified by SDS-polyacrylamide gel electrophoresis are 15,000 dalton,18,000 dalton and 35,000 dalton,respectively.展开更多
基金supported by the Priority Academic Program Development of Jiangsu Higher Education Institutions and the Industry-academic Joint Technological Innovations Funded Project of Jiangsu Province(BY2012172)
文摘Objective A subcutaneous transplantation tumor model of human HT-29 cells in nude mice was established to evaluate anticarcinogenic activities, and the apoptosis-regulated mechanism effect of aqueous extract of fermented wheat germ with Lactobacillus plantarum dy-1 (LFWGE). Methods The HT-29 cells were transplanted via subcutaneous injection of 1×10^7cells into the right flank of each nude mouse. Then, nude mice were treated for 30 d with LFWGE (high-dose 2 g/kg/d; low-dose 1 g/kg/d) and for 7 d with 5-fluorouracil (5-FU, 25 mg/kg/d) by gavage and intraperitoneal injection, respectively. An inhibition of tumor growth was observed. Results Tumor volume and weights decreased significantly in both groups of nude mice treated with LFWGE. In addition, the cell apoptosis rate of the LFWGE group (2 g/kg/d, 60.2%+4.4%; 1 g/kg/d, 58.6%+6.9%) was significantly higher than that of the control group (11.5%+1.6%) and 5-FU group (32.1%+3.5%) as measured by the TUNEL assay. Moreover, the real-time fluorescent quantitative PCR and Western blot method further confirmed these enhancing apoptosis and growth inhibition effects. The involvement of LFWGE in inducing apoptosis was confirmed by the expression of Bax, Bcl-2, Caspase-3, and CyclinD1. Conclusion The results showed that LFWGE could induce subcutaneous transplantation tumor apoptosis in nude mice and could be as a natural nutrient supplements or chemopreventive agent in the treatment of human colon cancer.
文摘An optical biosensor with a stirred cuvette has been used to monitor the interaction of immobilized wheat germ agglutinin (WGA) with two water-soluble cationic porphyrins, The association constants (Ka) of the free base porphyrin and its Zn(Ⅱ) complex form were 2. 66 and 27.31 × 10^5 l,/mol at 20 ℃ respectively. The interactions of the free base porphyrin were further investigated at temperatures between 15 ℃ and 37 ℃, The thermodynamics parameters, changes in free energy, enthalpy and entropy, were -31.23, 22.92, 54.15 ld/mol respectively. The heat capacity change was -355.53 J · mol^-1·K^-1 The binding was driven by entropic contribution, and showed strong enthalpy-entropy compensation. It was governed primarily by hydrophobic forces.
文摘[Objectives]This study was conducted to develop a health-care product with smooth taste and high nutrition.[Methods]With natural wheat germ powder as the main raw material and maltodextrin,white sugar,compound stabilizer and other materials as auxiliary materials,the formula of wheat germ solid beverage was optimized using sensory evaluation value as an index,through the mixture experiment method and fuzzy mathematical analysis.[Results]The optimal formula of wheat germ solid beverage was:sugar 11.51%,maltodextrin 37.89%,wheat germ powder 50%,compound stabilizer 0.6%(arabic gum 0.3%,xanthan gum 0.3%),soya bean lecithin 0.2%,ethyl maltol 0.002%,and vanillin 0.003%.[Conclusions]This study provides a theoretical basis for expanding the market-oriented production and application of wheat germ.
文摘Several studies are conducted to improve nutritional enrichment of ice cream. The aim of the present study was to produce healthy ice cream by partial replacement of fat with whey protein (WP), dietary fibers (oat (OT) and wheat germs (WG)) and modified starch (MS). Reduced fat ice cream (8% fat) was prepared as control. Healthy reduced fat ice cream was prepared by replacing 1% and 2% of fat with WP, OT, WG and MS. The fat replacement with OT and WG increased the fiber content of ice cream. In addition, ice cream from all treatments characterized by a significant great free radical scavenging activity when compared with control, especially when fat was replaced with WG, WP and OT, respectively. The sensory evaluation of ice cream revealed an enhancement of flavor when fat was replaced with 1% WP and OT and 2% WP (P < 0.05 vs. control). The modified starch treatments (1% and 2%) showed the highest viscosity followed by OT (2%) treatment (P < 0.05). In conclusion, these results showed that dietary fibers (OT & WG), WP and MS could be used to replace fat in ice cream and to give a healthy reduced fat ice cream with high fiber content and with high antioxidant activity.
文摘Wheat germ protein hydrolysates were prepared by protease hydrolysis, ultrafiltration and dynamical adsorption of resin. The total amount of amino acids in 100 g wheat germ protein hydrolysates is 93.95 g. Wheat germ protein hydrolysates are primarily composed of 4 fractions: 17.78% in the relative molecular mass range of 115631512, 17.50% in 1512842, 27.38% in 842372 and 30.65% in 37276, respectively. The antioxidant properties of wheat germ protein hydrolysates were evaluated by using different antioxidant tests in vitro. 1.20 g/L wheat germ protein hydrolysates exhibit 78.75% inhibition of peroxidation in linolei acid system; and 1.6g/L wheat germ protein hydrolysates show 81.11% scavenging effect on the 1,1-diphenyl-2-picrylhrazyl radical. The reducing power of 2.50 g/L wheat germ protein hydrolysates is 0.84. Furthermore, the scavenging activity of 0.60 g/L wheat germ protein hydrolysates against superoxide radical is 75.40%; 0.50 g/L wheat germ protein hydrolysates exhibit 63.35% chelating effect on ferrous ion. These antioxidant activities of wheat germ protein hydrolsates increase with the increase of its concentration. Experimental results suggest that wheat germ protein hydrolysate is a suitable natural antioxidant rich in nutrition and nontoxic.
文摘Wheat germ is a by-product derived from the wheat milling industry. Defatted wheat germ is the main by-product of the wheat germ in the oil extraction process. This study aims at development of efficient and low cost processing methods to transform these residues in added value co-product. In this study, wheat germ was analysed for its proximate composition, fatty acid composition, physical and chemical characteristics of wheat germ oil. The basic chemical composition analyses revealed high values of dry matter (87.37 g/100g FW), significant amounts of total protein and fat (27.69 and 8.99 g/100g FW, respectively) content and low ash content (3.08 g/100g FW). The quality of the extracted oils was assessed in terms of acid value, iodine value, saponification value, peroxide value, refractive index, and unsaponifiable matter. The fatty acid profile was found to be made up of linoleic followed by palmitic and oleic as the major fatty acids. Antioxidant properties and in vitro antibacterial activity of defatted wheat germ (DWG) extract were also determined. DWG, as a source of natural antioxidants and antibacterial, could be used to formulate nutraceuticals with potential applications to reduce the level of oxidative stress. The antioxidant potency of the DWG extracts could be the basis for its health promoting potential. The results showed that these by-products could be used as a source of bioactive compounds beneficial for health.
文摘This work aimed at determining the maximum acceptable level of wheat germ that can be added to one of homemade biscuits in Jordan, studying some chemical and sensory properties of this product, moreover, studying the effect of storage time after two months at room temperature on the sensory parameters of this product. The results showed that the addition of wheat germ in biscuit making at level of 20% (w/w) enhanced the nutritional value by increasing protein and total mineral content, as well as decreasing fat, total carbohydrates and total calories. The acceptability of sensory characteristics was improved by this fortification immediately after preparation and during storage.
文摘We describe a two-step method that uses wheat germ agglutinin immobilized on Sepharose gel followed by immunoaffinity chromatography (IAC) to extract recombinant human erythropoietin and Darbepoetin from equine plasma. Lectin affinity chromatography was shown to be an effective approach for isolating these epoetins from plasma and in combination with IAC;this method gave superior recovery when compared to the use of the latter technique alone. Moreover, due to the ease with which it can be scaled up, it is particularly well suited for pre-concentrating larger volumes of samples prior to IAC and this provides a facile way of improving the overall sensitivity with which these foreign proteins can be detected in equine plasma.
基金Punjab Agriculture Research Board funds for the project "A comprehensive integrated scientific approach for the development of sustainable management strategies of pink bollworm(Pectinophora gossypiella)".
文摘Background Pink bollworm,Pectinophora gossypiella(Saunders)(Lepidoptera:Gelechiidae)has become a poten-tial pest of cotton by causing substantial yield losses around the world including Pakistan.Keeping in view the facts like limited research investigations,unavailability,and high cost of artificial diet’s constituents and their premixes,the present research investigations on the dietary aspect of P.gossypiella were conducted.The larvae of P.gossypiella were reared on different diets that were prepared using indigenous elements.The standard/laboratory diet com-prised of wheat germ meal 34.5 g,casein 30.0 g,agar–agar 20.0 g,sucrose 10.0 g,brewer’s yeast 5.0 g,α-cellulose 1.0 g,potassium-sorbate1.5 g,niplagin 0.5 g,decavitamin 0.01 g,choline-chloride 0.06 g,maize-oil 3.30 g,honey 2.0 g,and water 730.0 mL.Alternatives to cotton bolls and wheat germ meal were okra seed sprouts,okra fruit,cottonseed meal,and okra seed meals,which were included in the study to introduce an efficient and economic mass-rearing system.Results The larval development completed in 19.68d±0.05 d with a weight of 20.18mg±0.20 mg at the fourth instar fed on the cottonseed meal-based diet instead of wheat germ meal based diet.On the same diet,84.00%±4.00%,17.24 mg±0.03 mg,and 7.76d±0.06 d were recorded as pupae formation,pupal weight,and pupal duration,respectively.Adult emergence,76.00%±1.00%was recorded from pupae collected from larvae raised on cottonseed meal-based diet.These male and female moths lived for 40.25d±0.10 d,and 44.34d±0.11 d,respectively.Females deposited 21.28±0.04 eggs per day with the viability of 65.78%±0.14%.The larval mortal-ity at the fourth instar was 37.20%±1.36%and malformed pupation of 12.00%±1.41%was recorded.Replacement of wheat germ meal with that of local meals(cottonseed and okra seed)in the standard laboratory diet has saved 463.80 to 467.10 PKR with 1.62 to 1.63 cost economic returns,respectively.Conclusion This research is of novel nature as it provides a concise and workable system for the economic and suc-cessful rearing of P.gossypiella under laboratory conditions.
文摘In order to refine the products of wheat germ agglutinin(WGA),several ion exchangers,such as D261,732,DEAE-cellulose-32 and CM-cellulose-32, have been used to removed proteins and pigments with different colors,such as brown,red,yellow,green and black,in the extracts of wheat germ.The WGA obtained from this procedure has higher hemagglutination activity than that available from sigma Co. The minimum hemagglutination dose of the purified WGA for an equal volume of a 2% type A red blood cells is 4μg/ml.WGA is a mixture of isolectins with different isoelectric points,5.9,6.2 and 6.8.Their molecular weight identified by SDS-polyacrylamide gel electrophoresis are 15,000 dalton,18,000 dalton and 35,000 dalton,respectively.