C-reactive protein to albumin ratio predict responses to programmed cell death-1 inhibitors in hepatocellular carcinoma patients

BACKGROUND Over the years,programmed cell death-1(PD-1)inhibitors have been routinely used for hepatocellular carcinoma(HCC)treatment and yielded improved survival outcomes.Nonetheless,significant heterogeneity surrounds the outcomes of most studies.Therefore,it is critical to search for biomarkers that predict the efficacy of PD-1 inhibitors in patients with HCC.AIM To investigate the role of the C-reactive protein to albumin ratio(CAR)in evaluating the efficacy of PD-1 inhibitors for HCC.METHODS The clinical data of 160 patients with HCC treated with PD-1 inhibitors from January 2018 to November 2022 at the First Affiliated Hospital of Guangxi Medical University were retrospectively analyzed.RESULTS The optimal cut-off value for CAR based on progression-free survival(PFS)was determined to be 1.20 using x-tile software.Cox proportional risk model was used to determine the factors affecting prognosis.Eastern Cooperative Oncology Group performance status[hazard ratio(HR)=1.754,95%confidence interval(95%CI)=1.045-2.944,P=0.033],CAR(HR=2.118,95%CI=1.057-4.243,P=0.034)and tumor number(HR=2.932,95%CI=1.246-6.897,P=0.014)were independent prognostic factors for overall survival.CAR(HR=2.730,95%CI=1.502-4.961,P=0.001),tumor number(HR=1.584,95%CI=1.003-2.500,P=0.048)and neutrophil to lymphocyte ratio(HR=1.120,95%CI=1.022-1.228,P=0.015)were independent prognostic factors for PFS.Two nomograms were constructed based on independent prognostic factors.The C-index index and calibration plots confirmed that the nomogram is a reliable risk prediction tool.The ROC curve and decision curve analysis confirmed that the nomogram has a good predictive effect as well as a net clinical benefit.CONCLUSION Overall,we reveal that the CAR is a potential predictor of short-and long-term prognosis in patients with HCC treated with PD-1 inhibitors.If further verified,CAR-based nomogram may increase the number of markers that predict individualized prognosis.

C-reactive Protein/Albumin Ratio as a Prognostic Indicator in Posttraumatic Shock and Outcome of Multiple Trauma Patients

Objective C-reactive protein(CRP)/albumin ratio(CAR)is a new inflammation-based index for predicting the prognosis of various diseases.The CAR determined on admission may help to predict the prognostic value of multiple trauma patients.Methods A total of 264 adult patients with severe multiple trauma were included for the present retrospective study,together with the collection of relevant clinical and laboratory data.CAR,CRP,albumin,shock index and ISS were incorporated into the prognostic model,and the receiver operating characteristic(ROC)curve was drawn.Then,the shock index for patients with different levels of CAR was analyzed.Finally,univariate and multivariate logistic regression analyses were performed to identify the independent risk factors for the 28-day mortality of multiple trauma patients.Results A total of 36 patients had poor survival outcomes,and the mortality rate reached 13.6%.Furthermore,after analyzing the shock index for patients with different levels of CAR,it was revealed that the shock index was significantly higher when CAR was≥4,when compared to CAR<2 and 2≤CAR<4,in multiple trauma patients.The multivariate logistic analysis helped to identify the independent association between the variables CAR(P=0.029)and shock index(P=0.019),and the 28-day mortality of multiple trauma patients.Conclusion CAR is higher in patients with severe multiple trauma.Furthermore,CAR serves as a risk factor for independently predicting the 28-day mortality of multiple trauma patients.The shock index was significantly higher when CAR was≥4 in multiple trauma patients.

The prognostic potential of pretreatment C-reactive protein to albumin ratio in stage IE/IIE extranodal natural killer/T-cell lymphoma

Objective The aim of this study was to determine the prognostic significance of the C-reactive protein-toalbumin ratio (CRP/Alb) for stage IE/IIE upper aerodigestive tract extranodal NK/T cell lymphoma patients. Methods One hundred and fourteen patients diagnosed with extranodal NK/T cell lymphoma at Sichuan Cancer Hospital from September 2011 to November 2016 were retrospectively reviewed. An optimal cutoff value of CRP/Alb for overall survival rate as an endpoint was obtained using the receiver operating curve (ROC). Results The optimal cutoff value of CRP/Alb was 0.15. For the low CRP/Alb group, the 3-year progression-free survival (PFS) was 78.6% and the 3-year overall survival (OS) was 80.7%. The 3-year PFS and OS values for the high CRP/Alb group were 41.6% and 45.2%, respectively. Differences for PFS (P < 0.001) and OS (P < 0.001) between the two groups were statistically significant. Univariate analysis showed that ECOG, IPI, CRP, GPS, and CRP/Alb were significantly associated with PFS. Similarly, all five were also significantly associated with OS. Multivariate analysis further confirmed that ECOG and CRP/ Alb were independent prognostic factors for both PFS and OS. Moreover, the cutoff value of CRP/Alb showed superior prognostic ability in discriminating between patients with different outcomes in low-risk group based on GPS, IPI, and KPI scores. Conclusion CRP/Alb is a promising prognostic marker for early-stage extranodal NK/T cell lymphoma.

Composition and Molecular Weight Distribution of Albumin and Globulin Protein Isolates from Durum Wheat Genotypes

This paper attempts to evaluate the banding patterns of non-gluten protein isolates from the grain of durum wheat varieties. Under reduced condition, polyacrylamide-gel electrophoresis has revealed a number of different sized albumin and globulin protein bands. The electrophoretic pattern of globulin showed more polymorphisms than that of albumin. High polymorphism, both in band intensity and occurrence, was observed between 15 kDa and 35 kDa. Most of the protein bands were scored in the range of 10 kDa and 85 kDa in the two protein fractions. At a cutoff point 2.5, cluster analysis based on the SDS-PAGE of globulin proteins classified the durum wheat varieties into three major family groups. Generally, the experiment showed the suitability and usefulness of globulin protein fractions as a genetic marker in discriminating durum wheat genotypes.

Z-Scan technique: To measure the total protein and albumin in blood

Z-scan technique is an effective tool for measuring the optical nonlinearity of the materials. By using this technique the measurement was made for total protein and albumin. The nonlinear refractive index of the total protein and albumin were found to vary linearly with concentration. Hence by calculating the nonlinear refractive index it is possible to measure their concentration in the sample. The values measured thus are found in equivalence with conventional colorimetric method.

Assessment of C-Reactive Protein/Serum Albumin Ratio in Relation to Acute Presentation and Early Outcome of Patients with Acute Coronary Syndrome

Background: Acute coronary syndrome (ACS) is the leading cardiovascular (CV) cause of mortality. C reactive protein (CRP) has linked with long-term risk of recurrent cardiovascular events or death. Albumin, in contrast to CRP known as a negative acute-phase protein. Thus a newly introduced marker assessed relation of CRP to albumin ratio (CAR), which may provide better results than the use of either marker alone. The aim of the study is to assess the association of C-reactive protein to albumin ratio (CAR) with in-hospital short-term major adverse cardiac events (MACEs) in acute coronary syndrome (ACS) patients. Patients & Methods: A multi-centers prospective cohort study was conducted at coronary intensive care units (CICU) in Baghdad during the period from March to October 2021 that included a total of 132 patients who were diagnosed as a case of ACS. They were assessed for major adverse cardiac events (MACEs) like cardiogenic shock, arrhythmias, post-MI angina, and acute heart failure while inside the ward, in addition to need for early interventional therapeutic approach in relation to (CAR) immediately at time of admission to hospital. Results: High values of CAR, whether using hs-CRP or CRP, were identified as an independent predictor for in-hospital MACEs (P value Conclusion: The CAR was independently correlated with in-hospital short-term MACEs and can be used for risk stratification in patients with ACS.

Heparin-binding protein combined with human serum albumin in early assessment of community-acquired pneumonia:A retrospective study

Objective:To investigate the application of heparin-binding protein along with albumin(HBP+ALB)in evaluating the severity of community-acquired pneumonia(CAP)and compares it with single HBP,white blood cells(WBCs),C-reactive protein(CRP),and procalcitonin(PCT).Methods:A total of 226 patients with CAP admitted to the Emergency Department of Zhujiang Hospital,Southern Medical University,Guangdong,China,between March 1,2021,and March 1,2022,were enrolled.The patients were grouped into two groups:mild CAP(n=175)and severe CAP(sCAP)(n=51).Patients'characteristics and laboratory data were obtained.ROC curve and the value of the area under the curve(AUC)were used to evaluate the predictive values of HBP,ALB,WBC,CRP,and PCT.Results:WBC count,CRP,PCT,HBP,creatinine,and D-dimer were higher in the sCAP group,while ALB was lower in the sCAP group(P<0.05)than those in the mild CAP group.The AUCs of WBC,CRP,PCT,HBP,and HBP+ALB were 0.633(95%CI:0.545-0.722,P<0.05),0.635(95%CI:0.542-0.729,P<0.05),0.705(95%CI:0.619-0.791,P<0.05),0.809(95%CI:0.736-0.883,P<0.05),and 0.889(95%CI:0.842-0.936,P<0.05),respectively.Conclusions:HBP+ALB has a higher predictive value than single HBP,PCT,CRP and WBC used alone for the early assessment of CAP.

Pressure- and Urea-Induced Denaturation of Bovine Serum Albumin: Considerations about Protein Heterogeneity

Urea denatures proteins at different concentrations, depending on the experimental conditions and the protein. We in-vestigated the pressure-induced denaturation of bovine serum albumin (BSA) in the presence of subdenaturing concen-trations of urea based on a two-state equilibrium. Pressure-induced denaturation was enhanced at urea concentrations ([U]) of 3.5 M to 8.0 M, with the free energy of denaturation at atmospheric pressure ranging from +5.0 to –2.5 kJ/mol of BSA. The m values appeared to be biphasic, with m1 and m2 of 0.92 and 2.35 kJ mol–1?M–1, respectively. Plots of versus ln[U] yielded values of u, the apparent stoichiometric coefficient, of 1.68 and 6.67 mol of urea/mol of BSA for m1 and m2, respectively. These values were compared with the m and u values of other monomeric proteins reported in or calculated from the literature. The very low values of u systematically observed for proteins were suggestive of heterogeneity in the free energy of denaturation. Thus, a u value of 140 mol of urea/mol of BSA may indicate the existence of a heterogeneous molecular population with respect to the free energy of dena-turation.

C反应蛋白/白蛋白比值对2型糖尿病合并急性心肌梗死患者远期不良心脑血管事件的预测价值研究

背景急性心肌梗死(AMI)是威胁全球公众健康的主要原因之一。虽然已有相应的再灌注治疗策略,但AMI相关的主要不良心脑血管事件(MACCEs)仍然是全世界人口死亡的原因之一。尤其合并糖尿病的AMI患者,因冠状动脉病变复杂,病变程度严重,尽早发现和判断该部分患者远期预后相对困难,因此寻找相对简便、易获得的实验室指标,有利于为2型糖尿病(T2DM)合并AMI患者经皮冠状动脉介入(PCI)术后MACCEs的预测提供依据。目的探讨血清C反应蛋白(CRP)/白蛋白(Alb)比值(CAR)对T2DM合并AMI患者PCI术后远期MACCEs的预测价值。方法纳入2014—2019年就诊于宁夏医科大学总医院心血管内科1683例T2DM合并AMI患者为研究对象,收集患者的一般临床资料与检查结果。对所有患者进行电话或门诊随访,以全因死亡、非致死性心肌梗死、再发不稳定型心绞痛、非致死性脑卒中、新发心力衰竭或心力衰竭加重再入院、再次血运重建作为MACCEs。根据患者随访期间是否发生MACCEs分为MACCEs组(508例)和非MACCEs组(1175例)。采用单因素及多因素Logistic回归分析探讨T2DM合并AMI患者MACCEs事件的影响因素。采用Kaplan-Meier法绘制患者的生存曲线,生存曲线的比较采用Log-rank检验。采用受试者工作特征(ROC)曲线分析CAR对T2DM合并AMI患者远期发生MACCEs的预测效能,使用净重分类改善指标(NRI)和综合判别指数(IDI)评价CAR对T2DM合并AMI患者预后评估的改善效果。结果1683例患者中508例(30.18%)患者发生MACCEs。多因素Logistic回归分析显示高血压病[OR(95%CI)=1.994(1.142~3.483)]、冠状动脉植入支架长度[OR(95%CI)=1.031(1.002~1.062)]、CRP[OR(95%CI)=0.950(0.915~0.986)]、Alb[OR(95%CI)=0.933(0.880~0.989)]及CAR[OR(95%CI)=5.582(1.705~18.277)]是T2DM合并AMI患者PCI术后发生MACCEs的影响因素(P<0.05)。根据CAR中位表达水平(0.86),将患者分为CAR<0.86组和CAR≥0.86组,Log-rank检验结果显示,CAR≥0.86组MACCEs发生率高于CAR<0.86组(52.68%与22.92%;χ^(2)=65.65,P<0.001)。ROC曲线显示CAR预测T2DM合并AMI患者发生MACCEs的ROC曲线下面积为0.728(95%CI=0.702~0.754),最佳截断值为0.576,灵敏度为0.617,特异度为0.747。在基线模型基础上,与CRP、Alb相比,CAR能明显改善对患者发生MACCEs的预测效果(NRI=0.377,IDI=0.166,C指数=0.690;P<0.05)。结论CAR是T2DM合并AMI患者PCI术后远期MACCEs发生风险的有效预测指标。

急性脑梗死患者Trevo支架取栓术后转归影响因素及血清PTX3、IMA水平变化分析

目的分析急性脑梗死(acutecerebralinfarction,ACI)患者Trevo支架取栓术后转归影响因素及血清正五聚体蛋白(PTX3)、缺血修饰白蛋白(IMA)水平变化。方法选取2020年11月至2023年11月在许昌医院行Trevo支架取栓治疗的150例ACI患者为研究对象,根据患者术后恢复情况分为预后优良组93例,预后不良组57例。比较两组一般资料及治疗前后血清PTX3、IMA水平,分析影响术后转归的因素及联合诊断预后转归的预测价值。结果两组年龄、发病至入院时间、梗死部位、收缩压连续变异性方面比较,差异具有统计学意义(P<0.05);治疗后预后优良组血清PTX3、IMA水平低于预后不良组(P<0.05);年龄、发病至入院时间、收缩压连续变异性为影响预后转归的不良因素(P<0.05);梗死部位为影响预后转归的保护因素(P<0.05);血清PTX3、IMA水平联合诊断ACI患者Trevo支架取栓术后恢复情况的AUC为0.913。结论ACI患者Trevo支架取栓术后转归与年龄、发病至入院时间、梗死部位、收缩压连续变异性密切相关,监测治疗前后血清PTX3、IMA水平可作为临床评估患者预后恢复情况的参考指标。

Citrate-stabilized CdSe/CdS quantum dots as fluorescence probe for protein determination

A rapid, ultrasensitive and convenient fluorescence measurement technology based on the enhancement of the fluorescence intensity resulting from the interaction of functionalized CdSe/CdS quantum dots （QDs） with bov/ne serum albumin （BSA） was proposed. The citrate-stabilized CdSe/CdS （QDs） were synthesized by using Se powder and Na2S as precursors instead of any pyrophoric organometallic precursors. The modified CdSe/CdS QDs are brighter and more stable against photobleaching in comparison with organic fluorophores. At pH 7.0, the fluorescence signal of CdSe/CdS is enhanced by increasing the concentration of BSA in the range of 0.1-10 μg/mL, and the low detection limit is 0.06 μg/mL. A linear relationship between the enhanced fluorescence peak intensity （△F） and BSA concentration （c） is established using equation △F=50.7c＋16.4 （R=0.996 36）. Results of determination for BSA in three synthetic samples are identical with the true values, and the recovery （98.9%-102.4%） and relative standard deviation （RSD, 1.8%-2.5%） are satisfactory.

Denaturation studies on bovine serum albumin–bile salt system:Bile salt stabilizes bovine serum albumin through hydrophobicity

Protein denaturation is under intensive research, since it leads to neurological disorders of severe consequences. Avoiding denaturation and stabilizing the proteins in their native state is of great importance,especially when proteins are used as drug molecules or vaccines. It is preferred to add pharmaceutical excipients in protein formulations to avoid denaturation and thereby stabilize them. The present study aimed at using bile salts(BSs), a group of well-known drug delivery systems, for stabilization of proteins.Bovine serum albumin(BSA) was taken as the model protein, whose association with two BSs, namely sodium cholate(Na C) and sodium deoxycholate(Na DC), was studied. Denaturation studies on the preformed BSA-BS systems were carried out under chemical and physical denaturation conditions. Urea was used as the chemical denaturant and BSA-BS systems were subjected to various temperature conditions to understand the thermal(physical) denaturation. With the denaturation conditions prescribed here,the data obtained is informative on the association of BSA-BS systems to be hydrophobic and this effect of hydrophobicity plays an important role in stabilizing the serum albumin in its native state under both chemical and thermal denaturation.

C-reactive protein as a predictor for cardiac events in Chinese elderly patients with coronary heart disease

Background and objective To assess the predictive value of C-reactive protein(CRP) for major adverse cardiac events and the association between CRP level and the coronary lesion morphology and extent in patients with coronary heart disease (CHD).Methods CRP was measured on admission in 177 consecutive elderly (age≥60 years) patients with CHD who underwent coronary angiography. Patients were divided into high CRP group (CRP≥3mg/L) and normal CRP group (CRP ＜3mg/L). The association between CRP levels and the coronary lesion features, including severity of stenosis (mild, moderate, severe), extent of lesion (diffused or nondiffused), eccentricity of the plaque (eccentric or non-eccentric) were analyzed. Patients were followed up for a mean of 8 months for the occurrences of major adverse cardiac events (MACE). Results Compared with patients in normal CRP group, patients in high CRP group were more frequently to have unstable angina, multi-vessel, diffuse, eccentric lesions, positive remodeling, and non-smooth plaques (P＜0.01). Kaplan-Meier analysis showed patients in high CRP group had a significantly lower MACE-free survival rate than patients in normal CRP group (Log-rank = 12.0, P＜0.01); Cox regression analysis indicated CRP level as an independent predictor for the occurrence of MACE (OR=3.16, P＜0.05) Conclusions High CRP level is associated with more extend, severe and eccentric coronary lesions and is an independent predictor for MACE in elderly patients with CHD.

Effects of proteins on magnesium degradation-static vs.dynamic conditions

The interaction between organic molecules and biomaterial surfaces determines the fate of biomaterials during their service life,which is also the research hotspots in the field of biomaterials.To understand the mechanism of protein interaction with magnesium(Mg)degradation,alloying elements,immersion time,protein concentration and surface conditions have been previously considered for the effect of proteins on Mg degradation.However,fluid flow,as one of the critical factors,drew little attention in this case.In the present study,the effect of bovine serum albumin(BSA)and fetal bovine serum(FBS)on Mg degradation was compared under static and dynamic conditions.The results revealed that both BSA and FBS slightly decreased the degradation rate of Mg in Hanks’balanced salt solution(HBSS)under static immersion due to the protein adsorption and the formation of a Ca/P-rich top layer on Mg surface,whereas under dynamic flow condition the degradation of Mg was significantly accelerated in the presence of BSA or FBS.The reasons seemed to stem from the weakened protein adsorption on Mg surface in this case and the dynamically enhanced interaction between proteins and ions/products in solutions,which largely weaken the combination of the top Ca/P-rich layer with the inner corrosion product layer.These results highlight the importance of testing conditions for Mg characterization in vitro and the synergistic effect between different parameters on Mg degradation.

Albumin resuscitation protects against traumatic/hemorrhagic shock-induced lung apoptosis in rats

Objective: To determine the effects of albumin administration on lung injury and apoptosis in traumatic/hemorrhagic shock (T/HS) rats. Methods: Studies were performed on an in vivo model of spontaneously breathing rats with induced T/HS; the rats were subjected to femur fracture, ischemia for 30 min, and reperfusion for 20 min with Ringer's lactate solution (RS) or 5% (w/v) albumin (ALB), and the left lower lobes of the lungs were resected. Results: Albumin administered during reperfusion markedly attenuated injury of the lung and decreased the concentration of lactic acid and the number of in situ TdT-mediated dUTP nick-end labelling (TUNEL)-positive cells. Moreover, immunohistochemistry performed 24 h after reperfusion revealed increases in the level of nuclear factor κB (NF-κB), and phosphorylated p38 mitogen-activated protein kinase (MAPK) in the albumin-untreated group was down-regulated by albumin treatment when compared with the sham rats. Conclusion: Resuscitation with albumin attenuates tissue injury and inhibits T/HS-induced apoptosis in the lung via the p38 MAPK signal transduction pathway that functions to stimulate the activation of NF-κB.

Fluorimetric study on the interaction between fluoresceinamine and bovine serum albumin

Fluorescence spectroscopy was employed to investigate the interaction between fluorophore fluoresceinamine(FA) and bovine serum albumin(BSA) under physiological conditions. In the mechanism discussion, it was proved that the fluorescence quenching of BSA by FA is a result of the formation of a BSA-FA complex. Fluorescence quenching constants were determined using the modified Stern-Volmer equation to provide a measure of the binding affinity between FA and BSA. The results of the thermodynamic parameters △G, △H, and △S at different temperatures indicated that several kinds of interactions, except for the electrostatic interactions play cooperative roles in BSA-FA association. Furthermore, the conformation of BSA upon interaction with FA was also studied by synchrotron fluorescence spectroscopy.

Binding of naturally occurring hydroxycinnamic acids to bovine serum albumin

Hydroxycinnamic acids (HCAs) possess numer- ous biological effects including antioxidant, antiallergic, antimicrobial, and immunomodulatory activities and due to these properties are widely used in folk medicine. Nevertheless, they can interact with protein molecules and cause some structural and functional changes. The possib- ility of HCAs binding to bovine serum albumin (BSA) under physiological conditions was inve- stigated by the UV-VIS absorption spectroscopy and fluorescence quenching method. Apart from rosmarinic acid, all tested HCAs quenched tryptophan fluorescence of BSA in the studied range of concentrations (0-20 μM) mainly by static quenching mechanism (formation of non- fluorescent HCA-BSA complexes). The binding constants, number of binding sites and free energy changes were determined. The binding affinities of HCAs were ranked in the order: chlorogenic acid > sinapic acid ≥ caffeic acid > ferulic acid > o-coumaric acid > p-coumaric acid ≥ m-coumaric acid, which was confirmed by spectral overlaps of BSA emission spectrum with absorption spectrum of HCA. All free energy changes possessed negative sign indicating the spontaneity of HCA-BSA interaction.

Competitive Adsorption between Bovine Serum Albumin and Collagen Observed by Atomic Force Microscope

Atomic force microscopy (AFM) was used to study the competitive adsorption betweenbovine serum albumin (BSA) and type Ⅰ collagen on hydrophilic and hydrophobic silicon wafers.BSA showed a grain shape and the type Ⅰ collagen displayed fibril-like molecules with relativelyhomogeneous height and width, characterized with clear twisting (helical formation). These AFMimages illustrated that quite a lot of type Ⅰ collagen appeared in the adsorption layer on hydrophilicsurface in a competitive adsorption state, but the adsorption of BSA was more preponderant than thatof type Ⅰ collagen on hydrophobic silicon wafer surface. The experiments showed that theinfluence of BSA on type Ⅰ collagen adsorption on hydrophilic surface was less than that onhydrophobic surface.

Mechanisms of the Plurality of Scorpaena Porcus L. Serum Albumin

The proteins, which bind albuminspecific dye Evans blue, are revealed in the low-molecular protein fraction of the blood serum from Scorpaena porcus L. and identified as serum albumin. They were represented by three bands in 2D-SDS-PAAG. MALDI-TOF-analysis revealed the fundamental similarity of the mass spec-trum of the fragments of tryptic cleavage of proteins with molecular weight 73 and 76 kDa. The role of du-plications and intragenic reconstructions in the creation of the plurality of scorpaena albumins is discussed.

APPLICATION OF NONIONIC TEMPERATURE SENSITIVE HYDROGEL FOR CONCENTRATION OF PROTEIN AQUEOUS SOLUTION

Six different N-alkyl substituted acrylarnide nonionic hydrogels were prepared and their swelling characteristics were measured. Poly N-isopropyl acrylamide (PNIPA) and poly N-n-propyl-acrylamide (PNNPA) temperature sensitive hydrogels were chosen as the nonionic temperature sensitive hydrogels for concentration of very dilute aqueous protein solution. The separation properties of PNIPA and PNNPA hydr0gels with different network dimensions were studied and the modification of the hydrogels was surveyed in order to decrease their surface adsorption of protein molecules. The experimental results of the concentration of BSA (Bovin serum albumin) dilute aqueous solution by hydroxylpropyl methacrylate (HPMA) copolymerized PNIPA hydrogel were given. The value and the limitation of concentration of dilute aqueous protein solution by this method was evaluated.