基于Doehlert Matrix的全贯流泵装置定转子进出流间隙优化设计

全贯流泵是一种新型的机电-水泵一体式贯流泵,然而其在运行时存在定转子间隙回流,扰乱了叶轮内的流场分布,导致泵装置产生能量损失、压力波动和噪声等问题,影响泵站的正常运行。通过数值模拟和模型试验研究全贯流泵装置定转子间隙流的水力特性,结合Doehlert Matrix设计-响应面优化法对其定转子进、出流间隙结构进行优化设计,揭示全贯流泵装置定转子进、出流间隙结构对泵装置性能的影响机理,并得到最终优化的折角式定转子进、出流间隙结构方案为:外侧延伸段长度t1为4.921r,外侧收缩段长度x1为0.624r,内侧延伸段长度t2为3.655r,内侧收缩段长度x2为1.6r(r为定转子间隙宽度),使得全贯流泵装置扬程和效率分别提升约10.3%和5.2%。

Hydromechanical characterization of gas transport amidst uncertainty for underground nuclear explosion detection

Given the challenge of definitively discriminating between chemical and nuclear explosions using seismic methods alone,surface detection of signature noble gas radioisotopes is considered a positive identification of underground nuclear explosions(UNEs).However,the migration of signature radionuclide gases between the nuclear cavity and surface is not well understood because complex processes are involved,including the generation of complex fracture networks,reactivation of natural fractures and faults,and thermo-hydro-mechanical-chemical(THMC)coupling of radionuclide gas transport in the subsurface.In this study,we provide an experimental investigation of hydro-mechanical(HM)coupling among gas flow,stress states,rock deformation,and rock damage using a unique multi-physics triaxial direct shear rock testing system.The testing system also features redundant gas pressure and flow rate measurements,well suited for parameter uncertainty quantification.Using porous tuff and tight granite samples that are relevant to historic UNE tests,we measured the Biot effective stress coefficient,rock matrix gas permeability,and fracture gas permeability at a range of pore pressure and stress conditions.The Biot effective stress coefficient varies from 0.69 to 1 for the tuff,whose porosity averages 35.3%±0.7%,while this coefficient varies from 0.51 to 0.78 for the tight granite(porosity<1%,perhaps an underestimate).Matrix gas permeability is strongly correlated to effective stress for the granite,but not for the porous tuff.Our experiments reveal the following key engineering implications on transport of radionuclide gases post a UNE event:(1)The porous tuff shows apparent fracture dilation or compression upon stress changes,which does not necessarily change the gas permeability;(2)The granite fracture permeability shows strong stress sensitivity and is positively related to shear displacement;and(3)Hydromechanical coupling among stress states,rock damage,and gas flow appears to be stronger in tight granite than in porous tuff.

Construction of a Computational Scheme for the Fuzzy HIV/AIDS Epidemic Model with a Nonlinear Saturated Incidence Rate

This work aimed to construct an epidemic model with fuzzy parameters.Since the classical epidemic model doesnot elaborate on the successful interaction of susceptible and infective people,the constructed fuzzy epidemicmodel discusses the more detailed versions of the interactions between infective and susceptible people.Thenext-generation matrix approach is employed to find the reproduction number of a deterministic model.Thesensitivity analysis and local stability analysis of the systemare also provided.For solving the fuzzy epidemic model,a numerical scheme is constructed which consists of three time levels.The numerical scheme has an advantage overthe existing forward Euler scheme for determining the conditions of getting the positive solution.The establishedscheme also has an advantage over existing non-standard finite difference methods in terms of order of accuracy.The stability of the scheme for the considered fuzzy model is also provided.From the plotted results,it can beobserved that susceptible people decay by rising interaction parameters.

Review on laser directed energy deposited aluminum alloys

Lightweight aluminum(Al)alloys have been widely used in frontier fields like aerospace and automotive industries,which attracts great interest in additive manufacturing(AM)to process high-value Al parts.As a mainstream AM technique,laser-directed energy deposition(LDED)shows good scalability to meet the requirements for large-format component manufacturing and repair.However,LDED Al alloys are highly challenging due to their inherent poor printability(e.g.low laser absorption,high oxidation sensitivity and cracking tendency).To further promote the development of LDED high-performance Al alloys,this review offers a deep understanding of the challenges and strategies to improve printability in LDED Al alloys.The porosity,cracking,distortion,inclusions,element evaporation and resultant inferior mechanical properties(worse than laser powder bed fusion)are the key challenges in LDED Al alloys.Processing parameter optimizations,in-situ alloy design,reinforcing particle addition and field assistance are the efficient approaches to improving the printability and performance of LDED Al alloys.The underlying correlations between processes,alloy innovation,characteristic microstructures,and achievable performances in LDED Al alloys are discussed.The benchmark mechanical properties and primary strengthening mechanism of LDED Al alloys are summarized.This review aims to provide a critical and in-depth evaluation of current progress in LDED Al alloys.Future opportunities and perspectives in LDED high-performance Al alloys are also outlined.

Deer antler stem cell niche: An interesting perspective

In recent years,there has been considerable exploration into methods aimed at enhancing the regenerative capacity of transplanted and/or tissue-resident cells.Biomaterials,in particular,have garnered significant interest for their potential to serve as natural scaffolds for cells.In this editorial,we provide commentary on the study by Wang et al,in a recently published issue of World J Stem Cells,which investigates the use of a decellularized xenogeneic extracellular matrix(ECM)derived from antler stem cells for repairing osteochondral defects in rat knee joints.Our focus lies specifically on the crucial role of biological scaffolds as a strategy for augmenting stem cell potential and regenerative capabilities,thanks to the establishment of a favorable microenvironment(niche).Stem cell differen-tiation heavily depends on exposure to intrinsic properties of the ECM,including its chemical and protein composition,as well as the mechanical forces it can generate.Collectively,these physicochemical cues contribute to a bio-instructive signaling environment that offers tissue-specific guidance for achieving effective repair and regeneration.The interest in mechanobiology,often conceptualized as a form of“structural memory”,is steadily gaining more validation and momen-tum,especially in light of findings such as these.

High quality repair of osteochondral defects in rats using the extracellular matrix of antler stem cells

BACKGROUND Cartilage defects are some of the most common causes of arthritis.Cartilage lesions caused by inflammation,trauma or degenerative disease normally result in osteochondral defects.Previous studies have shown that decellularized extracellular matrix(ECM)derived from autologous,allogenic,or xenogeneic mesenchymal stromal cells(MSCs)can effectively restore osteochondral integrity.AIM To determine whether the decellularized ECM of antler reserve mesenchymal cells(RMCs),a xenogeneic material from antler stem cells,is superior to the currently available treatments for osteochondral defects.METHODS We isolated the RMCs from a 60-d-old sika deer antler and cultured them in vitro to 70%confluence;50 mg/mL L-ascorbic acid was then added to the medium to stimulate ECM deposition.Decellularized sheets of adipocyte-derived MSCs(aMSCs)and antlerogenic periosteal cells(another type of antler stem cells)were used as the controls.Three weeks after ascorbic acid stimulation,the ECM sheets were harvested and applied to the osteochondral defects in rat knee joints.RESULTS The defects were successfully repaired by applying the ECM-sheets.The highest quality of repair was achieved in the RMC-ECM group both in vitro(including cell attachment and proliferation),and in vivo(including the simultaneous regeneration of well-vascularized subchondral bone and avascular articular hyaline cartilage integrated with surrounding native tissues).Notably,the antler-stem-cell-derived ECM(xenogeneic)performed better than the aMSC-ECM(allogenic),while the ECM of the active antler stem cells was superior to that of the quiescent antler stem cells.CONCLUSION Decellularized xenogeneic ECM derived from the antler stem cell,particularly the active form(RMC-ECM),can achieve high quality repair/reconstruction of osteochondral defects,suggesting that selection of decellularized ECM for such repair should be focused more on bioactivity rather than kinship.

Ink-structing the future of vascular tissue engineering:a review of the physiological bioink design

Three-dimensional(3D)printing and bioprinting have come into view for a plannable and standardizable generation of implantable tissue-engineered constructs that can substitute native tissues and organs.These tissue-engineered structures are intended to integrate with the patient’s body.Vascular tissue engineering(TE)is relevant in TE because it supports the sustained oxygenization and nutrition of all tissue-engineered constructs.Bioinks have a specific role,representingthenecessarymedium for printability and vascular cell growth.This review aims to understand the requirements for the design of vascular bioinks.First,an in-depth analysis of vascular cell interaction with their native environment must be gained.A physiological bioink suitable for a tissue-engineered vascular graft(TEVG)must not only ensure good printability but also induce cells to behave like in a native vascular vessel,including self-regenerative and growth functions.This review describes the general structure of vascular walls with wall-specific cell and extracellular matrix(ECM)components and biomechanical properties and functions.Furthermore,the physiological role of vascular ECM components for their interaction with vascular cells and the mode of interaction is introduced.Diverse currently available or imaginable bioinks are described from physiological matrix proteins to nonphysiologically occurring but natural chemical compounds useful for vascular bioprinting.The physiological performance of these bioinks is evaluated with regard to biomechanical properties postprinting,with a view to current animal studies of 3D printed vascular structures.Finally,the main challenges for further bioink development,suitable bioink components to create a self-assembly bioink concept,and future bioprinting strategies are outlined.These concepts are discussed in terms of their suitability to be part of a TEVG with a high potential for later clinical use.

Application of Feature, Event, and Process Methods to Leakage Scenario Development for Offshore CO_(2) Geological Storage

Offshore carbon dioxide(CO_(2)) geological storage(OCGS) represents a significant strategy for addressing climate change by curtailing greenhouse gas emissions. Nonetheless, the risk of CO_(2) leakage poses a substantial concern associated with this technology. This study introduces an innovative approach for establishing OCGS leakage scenarios, involving four pivotal stages, namely, interactive matrix establishment, risk matrix evaluation, cause–effect analysis, and scenario development, which has been implemented in the Pearl River Estuary Basin in China. The initial phase encompassed the establishment of an interaction matrix for OCGS systems based on features, events, and processes. Subsequent risk matrix evaluation and cause–effect analysis identified key system components, specifically CO_(2) injection and faults/features. Building upon this analysis, two leakage risk scenarios were successfully developed, accompanied by the corresponding mitigation measures. In addition, this study introduces the application of scenario development to risk assessment, including scenario numerical simulation and quantitative assessment. Overall, this research positively contributes to the sustainable development and safe operation of OCGS projects and holds potential for further refinement and broader application to diverse geographical environments and project requirements. This comprehensive study provides valuable insights into the establishment of OCGS leakage scenarios and demonstrates their practical application to risk assessment, laying the foundation for promoting the sustainable development and safe operation of ocean CO_(2) geological storage projects while proposing possibilities for future improvements and broader applications to different contexts.

Facilitated Prediction of Micropollutant Degradation via UV-AOPs in Various Waters by Combining Model Simulation and Portable Measurement

The degradation of micropollutants in water via ultraviolet(UV)-based advanced oxidation processes(AOPs)is strongly dependent on the water matrix.Various reactive radicals(RRs)formed in UV-AOPs have different reaction selectivities toward water matrices and degradation efficiencies for target micropollutants.Hence,process selection and optimization are crucial.This study developed a facilitated prediction method for the photon fluence-based rate constant for micropollutant degradation(K′_(p,MP))in various UV-AOPs by combining model simulation with portable measurement.Portable methods for measuring the scavenging capacities of the principal RRs(RRSCs)involved in UV-AOPs(i.e.,HO^(·),SO_(4)^(·-),and Cl^(·))using a mini-fluidic photoreaction system were proposed.The simulation models consisted of photochemical,quantitative structure–activity relationship,and radical concentration steady-state approximation models.The RRSCs were determined in eight test waters,and a higher RRSC was found to be associated with a more complex water matrix.Then,by taking sulfamethazine,caffeine,and carbamazepine as model micropollutants,the k′_(p,MP) values in various UV-AOPs were predicted and further verified experimentally.A lower k′_(p,MP) was found to be associated with a higher RRSC for a stronger RR competition;for example,k′_(p,MP) values of 130.9 and 332.5 m^(2) einstein^(–1),respectively,were obtained for carbamazepine degradation by UV/H_(2)O_(2) in the raw water(RRSC=9.47×10^(4) s^(-1))and sand-filtered effluent(RRSC=2.87×10^(4) s^(-1))of a drinking water treatment plant.The developed method facilitates process selection and optimization for UV-AOPs,which is essential for increasing the efficiency and cost-effectiveness of water treatment.

Stiffness-tunable biomaterials provide a good extracellular matrix environment for axon growth and regeneration

Neuronal growth, extension, branching, and formation of neural networks are markedly influenced by the extracellular matrix—a complex network composed of proteins and carbohydrates secreted by cells. In addition to providing physical support for cells, the extracellular matrix also conveys critical mechanical stiffness cues. During the development of the nervous system, extracellular matrix stiffness plays a central role in guiding neuronal growth, particularly in the context of axonal extension, which is crucial for the formation of neural networks. In neural tissue engineering, manipulation of biomaterial stiffness is a promising strategy to provide a permissive environment for the repair and regeneration of injured nervous tissue. Recent research has fine-tuned synthetic biomaterials to fabricate scaffolds that closely replicate the stiffness profiles observed in the nervous system. In this review, we highlight the molecular mechanisms by which extracellular matrix stiffness regulates axonal growth and regeneration. We highlight the progress made in the development of stiffness-tunable biomaterials to emulate in vivo extracellular matrix environments, with an emphasis on their application in neural repair and regeneration, along with a discussion of the current limitations and future prospects. The exploration and optimization of the stiffness-tunable biomaterials has the potential to markedly advance the development of neural tissue engineering.

An approximate analytical model for unconventional reservoir considering variable matrix blocks and simultaneous matrix depletion

In regard to unconventional oil reservoirs,the transient dual-porosity and triple-porosity models have been adopted to describe the fluid flow in the complex fracture network.It has been proven to cause inaccurate production evaluations because of the absence of matrix-macrofracture communication.In addition,most of the existing models are solved analytically based on Laplace transform and numerical inversion.Hence,an approximate analytical solution is derived directly in real-time space considering variable matrix blocks and simultaneous matrix depletion.To simplify the derivation,the simultaneous matrix depletion is divided into two parts:one part feeding the macrofractures and the other part feeding the microfractures.Then,a series of partial differential equations(PDEs)describing the transient flow and boundary conditions are constructed and solved analytically by integration.Finally,a relationship between oil rate and production time in real-time space is obtained.The new model is verified against classical analytical models.When the microfracture system and matrix-macrofracture communication is neglected,the result of the new model agrees with those obtained with the dual-porosity and triple-porosity model,respectively.Certainly,the new model also has an excellent agreement with the numerical model.The model is then applied to two actual tight oil wells completed in western Canada sedimentary basin.After identifying the flow regime,the solution suitably matches the field production data,and the model parameters are determined.Through these output parameters,we can accurately forecast the production and even estimate the petrophysical properties.

Association between Gene Polymorphisms and SNP-SNP Interactions of the Matrix Metalloproteinase 2 Signaling Pathway and the Risk of Vascular Senescence

Objective This study aimed to explore the association of single nucleotide polymorphisms(SNP)in the matrix metalloproteinase 2(MMP-2)signaling pathway and the risk of vascular senescence(VS).Methods In this cross-sectional study,between May and November 2022,peripheral venous blood of151 VS patients(case group)and 233 volunteers(control group)were collected.Fourteen SNPs were identified in five genes encoding the components of the MMP-2 signaling pathway,assessed through carotid-femoral pulse wave velocity(cf PWV),and analyzed using multivariate logistic regression.The multigene influence on the risk of VS was assessed using multifactor dimensionality reduction(MDR)and generalized multifactor dimensionality regression(GMDR)modeling.Results Within the multivariate logistic regression models,four SNPs were screened to have significant associations with VS:chemokine(C-C motif)ligand 2(CCL2)rs4586,MMP2 rs14070,MMP2rs7201,and MMP2 rs1053605.Carriers of the T/C genotype of MMP2 rs14070 had a 2.17-fold increased risk of developing VS compared with those of the C/C genotype,and those of the T/T genotype had a19.375-fold increased risk.CCL2 rs4586 and MMP-2 rs14070 exhibited the most significant interactions.Conclusion CCL2 rs4586,MMP-2 rs14070,MMP-2 rs7201,and MMP-2 rs1053605 polymorphisms were significantly associated with the risk of VS.

MgO-attached graphene nanosheet(MgO@GNS)reinforced magnesium matrix nanocomposite with superior mechanical,corrosion and biological performance

Magnesium(Mg)alloys are gaining great consideration as body implant materials due to their high biodegradability and biocompatibility.However,they suffer from low corrosion resistance and antibacterial activity.In this research,semi-powder metallurgy followed by hot extrusion was utilized to produce the magnesium oxide@graphene nanosheets/magnesium(MgO@GNS/Mg)composite to improve mechanical,corrosion and cytocompatibility characteristics.Investigations have revealed that the incorporation of MgO@GNS nanohybrids into Mg-based composite enhanced microhardness and compressive strength.In vitro,osteoblast cell culture tests show that using MgO@GNS nanohybrid fillers enhances osteoblast adhesion and apatite mineralization.The presence of MgO@GNS nanoparticles in the composites decreased the opening defects,micro-cracks and micro-pores of the composites thus preventing the penetration of the corrosive solution into the matrix.Studies demonstrated that the MgO@GNS/Mg composite possesses excellent antibacterial properties because of the combination of the release of MgO and physical damage to bacterium membranes caused by the sharp edges of graphene nanosheets that can effectively damage the cell wall thereby facilitating penetration into the bacterial lipid bilayer.Therefore,the MgO@GNS/Mg composite with high mechanical strength,antibacterial activity and corrosion resistance is considered to be a promising material for load-bearing implant applications.

Periodic signal extraction of GNSS height time series based on adaptive singular spectrum analysis

Singular spectrum analysis is widely used in geodetic time series analysis.However,when extracting time-varying periodic signals from a large number of Global Navigation Satellite System(GNSS)time series,the selection of appropriate embedding window size and principal components makes this method cumbersome and inefficient.To improve the efficiency and accuracy of singular spectrum analysis,this paper proposes an adaptive singular spectrum analysis method by combining spectrum analysis with a new trace matrix.The running time and correlation analysis indicate that the proposed method can adaptively set the embedding window size to extract the time-varying periodic signals from GNSS time series,and the extraction efficiency of a single time series is six times that of singular spectrum analysis.The method is also accurate and more suitable for time-varying periodic signal analysis of global GNSS sites.

Microwave irradiation-induced alterations in physicochemical properties and methane adsorption capability of coals:An experimental study using carbon molecular sieve

In order to comprehend the applicability of microwave irradiation for recovering coalbed methane,it is necessary to evaluate the microwave irradiation-induced alterations in coals with varying levels of metamorphism.In this work,the carbon molecular sieve combined with KMnO_(4)oxidation was selected to fabricate carbon molecular sieve with diverse oxidation degrees,which can serve as model substances toward coals.Afterwards,the microwave irradiation dependences of pores,functional groups,and highpressure methane adsorption characteristics of model substances were studied.The results indicated that microwave irradiation causes rearrangement of oxygen-containing functional groups,which could block the micropores with a size of 0.40-0.60 nm in carbon molecular sieve;meanwhile,naphthalene and phenanthrene generated by macro-molecular structure pyrolysis due to microwave irradiation could block the micropores with a size of 0.70-0.90 nm.These alterations in micropore structure weaken the saturated methane adsorption capacity of oxidized carbon molecular sieve by 2.91%-23.28%,suggesting that microwave irradiation could promote methane desorption.Moreover,the increased mesopores found for oxidized carbon molecular sieve after microwave irradiation could benefit CH4 diffusion.In summary,the oxidized carbon molecular sieve can act as model substances toward coals with different ranks.Additionally,microwave irradiation is a promising technology to enhance coalbed methane recovery.

Single-cell manipulation by two-dimensional micropatterning

Cells are highly sensitive to their geometrical and mechanical microenvironment that directly regulate cell shape,cytoskeleton and organelle,as well as the nucleus morphology and genetic expression.The emerging two-dimensional micropatterning techniques offer powerful tools to construct controllable and well-organized microenvironment for single-cell level investigations with qualitative analysis,cellular standardization,and in vivo environment mimicking.Here,we provide an overview of the basic principle and characteristics of the two most widely-used micropatterning techniques,including photolithographic micropatterning and soft lithography micropatterning.Moreover,we summarize the application of micropatterning technique in controlling cytoskeleton,cell migration,nucleus and gene expression,as well as intercellular communication.

Biofabrication of nanocomposite-based scaffolds containing human bone extracellularmatrix for the differentiation of skeletal stem and progenitor cells

Autograft or metal implants are routinely used in skeletal repair.However,they fail to provide long-term clinical resolution,necessitating a functional biomimetic tissue engineering alternative.The use of native human bone tissue for synthesizing a biomimeticmaterial inkfor three-dimensional(3D)bioprintingof skeletal tissueis anattractivestrategyfor tissueregeneration.Thus,human bone extracellular matrix(bone-ECM)offers an exciting potential for the development of an appropriate microenvironment for human bone marrow stromal cells(HBMSCs)to proliferate and differentiate along the osteogenic lineage.In this study,we engineered a novel material ink(LAB)by blending human bone-ECM(B)with nanoclay(L,Laponite®)and alginate(A)polymers using extrusion-based deposition.The inclusion of the nanofiller and polymeric material increased the rheology,printability,and drug retention properties and,critically,the preservation of HBMSCs viability upon printing.The composite of human bone-ECM-based 3D constructs containing vascular endothelial growth factor(VEGF)enhanced vascularization after implantation in an ex vivo chick chorioallantoic membrane(CAM)model.The inclusion of bone morphogenetic protein-2(BMP-2)with the HBMSCs further enhanced vascularization and mineralization after only seven days.This study demonstrates the synergistic combination of nanoclay with biomimetic materials(alginate and bone-ECM)to support the formation of osteogenic tissue both in vitro and ex vivo and offers a promising novel 3D bioprinting approach to personalized skeletal tissue repair.

A method to predict rockburst using temporal trend test and its application

Rockbursts have become a significant hazard in underground mining,underscoring the need for a robust early warning model to ensure safety management.This study presents a novel approach for rockburst prediction,integrating the Mann-Kendall trend test(MKT)and multi-indices fusion to enable real-time and quantitative assessment of rockburst hazards.The methodology employed in this study involves the development of a comprehensive precursory index library for rockbursts.The MKT is then applied to analyze the real-time trend of each index,with adherence to rockburst characterization laws serving as the warning criterion.By employing a confusion matrix,the warning effectiveness of each index is assessed,enabling index preference determination.Ultimately,the integrated rockburst hazard index Q is derived through data fusion.The results demonstrate that the proposed model achieves a warning effectiveness of 0.563 for Q,surpassing the performance of any individual index.Moreover,the model’s adaptability and scalability are enhanced through periodic updates driven by actual field monitoring data,making it suitable for complex underground working environments.By providing an efficient and accurate basis for decision-making,the proposed model holds great potential for the prevention and control of rockbursts.It offers a valuable tool for enhancing safety measures in underground mining operations.

REFINEMENTS OF THE NORM OF TWO ORTHOGONAL PROJECTIONS

In this paper,some refinements of norm equalities and inequalities of combination of two orthogonal projections are established.We use certain norm inequalities for positive contraction operator to establish norm inequalities for combination of orthogonal projections on a Hilbert space.Furthermore,we give necessary and sufficient conditions under which the norm of the above combination of o`rthogonal projections attains its optimal value.

Amino-functionalized UiO-66-doped mixed matrix membranes with high permeation performance and fouling resistance

For the reduction of bovine serum proteins from wastewater,a novel mixed matrix membrane was prepared by functionalizing the substrate material polyaryletherketone(PAEK),followed by carboxyl groups(C-SPAEKS),and then adding amino-functionalized UiO-66-NH_(2)(Am-UiO-66-NH_(2)).Aminofunctionalization of UiO-66 was accomplished by melamine,followed by an amidation reaction to immobilize Am-UiO-66-NH_(2),which was immobilized on the surface of the membrane as well as in the pore channels,which enhanced the hydrophilicity of the membrane surface while increasing the negative potential of the membrane surface.This nanoparticle-loaded ultrafiltration membrane has good permeation performance,with a pure water flux of up to 482.3 L·m^(-2)·h^(-1) for C-SPAEKS/AmUiO-66-NH_(2) and a retention rate of up to 98.7%for bovine serum albumin(BSA)-contaminated solutions.Meanwhile,after several hydrophilic modifications,the flux recovery of BSA contaminants by this series of membranes increased from 56.2%to 80.55%of pure membranes.The results of ultra-filtration flux time tests performed at room temperature showed that the series of ultrafiltration membranes remained relatively stable over a test time of 300 min.Thus,the newly developed mixed matrix membrane showed potential for high efficiency and stability in wastewater treatment containing bovine serum proteins.