Existing therapies for the treatment of chronic heart failure still have some limitations and there is a pressing need for the development of new therapeutic modalities. The amniotic membrane has been used for the tre...Existing therapies for the treatment of chronic heart failure still have some limitations and there is a pressing need for the development of new therapeutic modalities. The amniotic membrane has been used for the treatment of various diseases, such as conjunctive defects;however, the mechanisms behind its repair functions are still unclear. Regenerative medicine is seeking newer alternatives and among them, biomaterials have emerged in recent years for developing and manipulating molecules, cells, tissues or organs grown in laboratories in order to replace human body parts. Many such materials have been used for this purpose, either synthetically or biologically, in order to provide new medical devices. This review provides a wider view of the regeneration potential of the use of amniotic membrane as a potential biomaterial to facilitate the implementation of new research in surgical procedures. Amniotic membrane appears to be an alternative source of stem cells as well as an excellent biomaterial for cell-based therapeutic applications in engineering heart tissue.展开更多
The combined inoculation of yeast and lactic acid bacteria(LAB)is a promising approach to enhance microbial metabo-lism and the quality of fermented food products.In this study,we investigated the effects of simultane...The combined inoculation of yeast and lactic acid bacteria(LAB)is a promising approach to enhance microbial metabo-lism and the quality of fermented food products.In this study,we investigated the effects of simultaneous and sequential inoculation of LAB and yeast on coffee fermentation.Fermentations were conducted using single and combined protocols with P.fermentans YC5.2 and P.pentosaceus LPBF07.A temporal analysis utilizing Illumina high-throughput rRNA Gene Sequencing revealed a synergistic interaction between the two microbial groups.This positive synergy led to increased consumption of coffee pulp sugar and the production of metabolites,surpassing the results observed in single cultures and the spontaneous process.Furthermore,the combined inoculation processes demonstrated a more significant role in sup-pressing wild microbiota compared to single cultures.Notably,the sequential process emerged as particularly effective in promoting a more intricate aroma profile and elevated sensorial score,attributed to the formation of distinctive compounds such as benzeneacetaldehyde,1-hexanol,2-heptanol,benzyl alcohol and phenylethyl alcohol.These results suggest that implementing a sequential inoculation process could enable coffee farmers to standardize on-farm processing and produce high-value-added products.展开更多
Interest in the technology for producing alternative proteins is rapidly increasing,driven by the need to find new ways to produce and consume protein for the global population.This technology involves growing differe...Interest in the technology for producing alternative proteins is rapidly increasing,driven by the need to find new ways to produce and consume protein for the global population.This technology involves growing different microorganisms and animal cells under controlled conditions to ensure their viability and efficient growth.The cultivation process takes place in different types of bioreactors,from traditional models to innovative new designs,each offering unique features and capabilities.The most commonly used bioreactors are stirred tank reactors,which are mechanically agitated,and airlift or bubble column bioreactors,which are pneumatically agitated.These bioreactors are often adapted or modified to optimize the production of cultured meat.Essential to the process are microcarriers or scaffolds that support cell adhesion and proliferation.Other bioreactor models,such as hollow fiber and packed bed bioreactors,are also being explored.The trend towards single-use technology is growing due to benefits like easier cleaning and sterilization,and reduced operation times,though it does raise concerns about plastic waste.This review not only describes various bioreactor models but also discusses instrumentation and control systems.It aims to present the main bioreactor models currently in use for cultivated meat production,detailing their features,advantages,disadvantages,and the technological challenges that need to be addressed.展开更多
Wet granulation-a unit operation involving mixing polymeric binders with powdered formulations-is well established in the pharmaceutical industry,playing a major role in the manufacturing of oral solid dosage forms an...Wet granulation-a unit operation involving mixing polymeric binders with powdered formulations-is well established in the pharmaceutical industry,playing a major role in the manufacturing of oral solid dosage forms and improving the physical properties of granules(size,density,shape factor,etc.)before tableting.The foaming properties of aqueous polymeric binders prove useful for binder delivery within the mixing vessel,with foamed binders leading to enhanced process efficiency(binder distribution,drying time,and temperature)and product quality(heat-sensitive components)during granulation.Given the importance of this stage in producing oral solid dosage forms,understanding the relationship between critical process parameters and critical quality attributes is essential.The process analytical technology(PAT)framework enables process design,analysis,and control and facilitates process development via in-line spectroscopy combined with multivariate data analysis to yield critical product information during the unit operation.Herein,we used in-line NIR spectroscopy to monitor granule size in foam granulations of a pharmaceutical compound.The mean granule diameter was predicted using a partial least squares regression(PLSR)model(with a prediction error of 11.8μm)and combined with a batch statistical process control(BSPC)approach for the temporal monitoring of granule size during three foam granulations.展开更多
The expanding field of alternative proteins represents a transformative approach to addressing global food security and sustainability challenges.Among these,fermentation-derived alternative proteins cultivated from m...The expanding field of alternative proteins represents a transformative approach to addressing global food security and sustainability challenges.Among these,fermentation-derived alternative proteins cultivated from microorganisms such as fungi,bacteria,and algae offer a promising avenue for sustainable protein production.This review explores the selection and utilization of raw materials to produce microbial proteins through fermentation processes.Critical raw materials include agricultural byproducts,industrial waste streams,and specifically designed feedstocks,which not only mitigate environmental footprint but also enhance the economic viability of production systems.The utilization of lignocellulosic biomass and molasses has demonstrated considerable promise,attributed to their abundant and renewable nature.The review underscored the necessity of exploring specific areas to enhance the viability of producing microbial protein from diverse raw materials.These areas include improving pre-treatment strategies to enhance substrate suitability for fermentation,optimizing fermentation processes for increased yield and reduced costs,and developing more resilient microorganisms capable of thriving on varied substrates.These strategies are crucial for advancing the production of alternative proteins through fermentation,in addition to raw material selection,which is vital in the scalability and sustainability of alternative protein production through fermentation,emphasizing the need for continued research and innovation in this field.展开更多
Enzymes play a pivotal role in meat processing and maturation,starting with endogenous enzymes that catalyse a partial proteolysis in natural meat that enhances texture and flavor,to exogenous enzymes that may be inco...Enzymes play a pivotal role in meat processing and maturation,starting with endogenous enzymes that catalyse a partial proteolysis in natural meat that enhances texture and flavor,to exogenous enzymes that may be incorporated for additional processing.Therefore,a question is raised whether endogenous and exogenous proteases could be used to improve the quality of meat products originated from in vitro cell-based technology.This manuscript aims at presenting the main applications of enzymes in meat processing,starting with conventional meat products and expanding to reported and potential applications in cultured meat technologies.Enzymes are applied since the early stages of cell isolation,multiplication and detaching from supports.The use of endogenous enzymes,such as cathepsins and calpains,has been proposed to improve the palatability of cultivated meat,and the application of transglutaminases has already been reported in scientific papers and patent documents,with the aim of promoting crosslinking reactions among proteins.This feature can be used to modulate the texture of cultivated meat products.Also,their use has been directed to the manufacture of scaffolds and microcarriers for animal cell cultivation.As concluded from the scientific and technological literature survey,it is evident that a substantial area of study remains unexplored regarding the utilization of enzymes in cultivated meat products.展开更多
Identifying alternative protein sources is crucial in view of the shortage of protein resources.A new strain Geotrichum can-didum IBB69 was isolated for microbial protein production in this study.The protein yield,bio...Identifying alternative protein sources is crucial in view of the shortage of protein resources.A new strain Geotrichum can-didum IBB69 was isolated for microbial protein production in this study.The protein yield,biomass,and specific protein production(the ratio of total intracellular and extracellular protein to biomass)of G.candidum IBB69 were 7.6 g/L,18.42 g/L and 60.8%after optimization through shaking flask fermentation,response surface methodology,and 5L scale-up fermen-tation.Compared to flask fermentation,the protein yield and content increased by 83.13%and 91.08%,respectively.The protein product of IBB69 was composed of 18 amino acids with a ratio of 37.51%(EAA to TAA).Transcriptome analysis revealed that,compared with ammonium sulfate,the addition of urea upregulated the expression levels of key genes in the carbon-nitrogen metabolism process of G.candidum IBB69,promoting amino acid synthesis and cell growth.This study tapped a new microbial protein producing strain,and optimized results set the stage for the industrial development and application of G.candidum for sustainable alternative proteins.展开更多
在基于核磁共振(NMR)的代谢组学数据分析中,尺度缩放是关键的预处理步骤之一,其主要目的是通过调整数据的方差结构,改善后续的多变量统计分析的结果。从信息熵的角度出发,利用KullbackLeibler(K-L)散度来度量不同实验分组的生物样品的1 ...在基于核磁共振(NMR)的代谢组学数据分析中,尺度缩放是关键的预处理步骤之一,其主要目的是通过调整数据的方差结构,改善后续的多变量统计分析的结果。从信息熵的角度出发,利用KullbackLeibler(K-L)散度来度量不同实验分组的生物样品的1 H NMR波谱数据的差异程度,并结合单位方差缩放法,提出一种基于K-L散度的尺度缩放方法。该方法先利用单位方差法将数据各变量的标准差调整到同一水平上,再利用K-L散度对各变量进行有监督地加权,增强重要变量、减弱无关变量。由于K-L散度是在概率分布的意义上度量数据间的差异程度,且对于高斯和非高斯分布的数据均适用,因此能更准确地度量不同实验分组样品的1 H NMR波谱数据的差异性,从而更有效地地对谱数据的重要变量进行识别和加权。人群尿液1 H NMR波谱数据的分析结果表明,基于K-L散度的尺度缩放方法能有效抑制噪声变量,同时很好地区分特征变量和非特征变量;提高主成分回归(PCR)模型的判别能力;改善偏最小二乘回归判别分析(PLS-DA)模型的解释能力、预测能力以及对特征代谢物的辨识能力。展开更多
Methylprednisolone is a commonly used drug for the treatment of spinal cord injury, but high doses of methylprednisolone can increase the incidence of infectious diseases. Methotrexate has anti-inflammatory activity a...Methylprednisolone is a commonly used drug for the treatment of spinal cord injury, but high doses of methylprednisolone can increase the incidence of infectious diseases. Methotrexate has anti-inflammatory activity and immunosuppressive effects, and can reduce in- flammation after spinal cord injury. To analyze gene expression changes and the molecular mechanism of methotrexate combined with methylprednisolone in the treatment of spinal cord injury, a rat model of spinal cord contusion was prepared using the PinPointTM preci- sion cortical impactor technique. Rats were injected with methylprednisolone 30 mg/kg 30 minutes after injury, and then subcutaneously injected with 0.3 mg/kg methotrexate 1 day after injury, once a day, for 2 weeks. TreadScan gait analysis found that at 4 and 8 weeks after injury, methotrexate combined with methylprednisolone significantly improved hind limb swing time, stride time, minimum longitudinal deviation, instant speed, footprint area and regularity index. Solexa high-throughput sequencing was used to analyze differential gene ex- pression. Compared with methylprednisolone alone, differential expression of 316 genes was detected in injured spinal cord treated with methotrexate and methylprednisolone. The 275 up-regulated genes were mainly related to nerve recovery, anti-oxidative, anti-inflammatory and anti-apoptotic functions, while 41 down-regulated genes were mainly related to proinflammatory and pro-apoptotic functions. These results indicate that methotrexate combined with methylprednisolone exhibited better effects on inhibiting the activity of inflammatory cytokines and enhancing antioxidant and anti-apoptotic effects and thereby produced stronger neuroprotective effects than methotrexate alone. The 316 differentially expressed genes play an important role in the above processes.展开更多
Particles occur in almost all processes in chemical and life sciences. The particle size and shape influence the process performance and product quality, and in turn they are influenced by the flow behavior of the par...Particles occur in almost all processes in chemical and life sciences. The particle size and shape influence the process performance and product quality, and in turn they are influenced by the flow behavior of the particles during production. Monitoring and controlling such characteristics in multiphase systems to obtain sufficient qualities will greatly facilitate the achievement of reproducible and defined distributions. So far, obtaining this information inline has been challenging, because existing instruments lack measurement precision, being unable to process overlapping signals from different particle phases in highly concentrated multiphase systems. However, recent advances in photo-optics made it possible to monitor such features(particle size distribution(PSD), aspect ratio and particle concentration) with advanced image analysis(IA) in real-time. New analysis workflows as well as single feature extractions from the images using multiple image analysis algorithms allowed the precise real-time measurements of size, shape and concentration of particle collectives even separated from each other in three phase systems. The performances, advantages and drawbacks with other non-photo-optical methods for assessing the particle size distribution are compared and discussed.展开更多
Ferrite/carbon composited materials,especially the bio-derived composited materials possessing both environmental friendliness and outstanding microwave absorption performance,attract numerous attentions for solving t...Ferrite/carbon composited materials,especially the bio-derived composited materials possessing both environmental friendliness and outstanding microwave absorption performance,attract numerous attentions for solving the"electromagnetic problem"in the Gigahertz frequency range.In this work,we demonstrate a bio-derived ferrite/carbon material by compositing functional carbonized cotton fibers(CCFs)and Fe_(3)O_(4)nanoparticles with optimized microwave-absorption properties.By adjusting the carbonization conditions systematically,the Fe_(3)O_(4)loading contents and the microwave absorption properties can be varied simultaneously-and,indeed,optimized and tuned.The CCFs-Fe_(3)O_(4)composites exhibited a minimum reflection-loss capacity RL(d B)of-56.8 d B at 10.9 GHz with a thickness of 1.67 mm,and its effective absorption bandwidth(RL(d B)<-20 d B)was found to broaden to 7.1 GHz.Electromagnetic characterizations,coupled with microstructure analyses,revealed that the enhancement in microwave absorption was triggered by the different microstructures of CCFs-Fe_(3)O_(4)composites-attributable to the different carbonization processes.These different conditions result in different amounts of Fe_(3)O_(4)attachment sites and lead to the enhancement of dielectric polarization at localized microstructures.The present work of bio-derived ferrite/carbon materials has important implications in understanding structure-performance relationships in dielectric-magnetic materials,and,meanwhile,could well be extended to a microwave-absorber design approach.展开更多
Ensuring the removal of host cell proteins(HCPs) during downstream processing of recombinant proteins such as monoclonal antibodies(m Abs) remains a challenge.Since residual HCPs might affect product stability or safe...Ensuring the removal of host cell proteins(HCPs) during downstream processing of recombinant proteins such as monoclonal antibodies(m Abs) remains a challenge.Since residual HCPs might affect product stability or safety,constant monitoring is required to demonstrate their removal to be below the regulatory accepted level of 100 ng/mg.The current standard analytical approach for this procedure is based on ELISA;however,this approach only measures the overall HCP content.Therefore,the use of orthogonal methods,such as liquid chromatography-mass spectrometry(LC-MS),has been established,as it facilitates the quantitation of total HCPs as well as the identification and quantitation of the individual HCPs present.In the present study,a workflow for HCP detection and quantitation using an automated magnetic bead-based sample preparation,in combination with a data-independent acquisition(DIA) LC-MS analysis,was established.Employing the same instrumental setup commonly used for peptide mapping analysis of m Abs allows for its quick and easy implementation into pre-existing workflows,avoiding the need for dedicated instrumentation or personnel.Thereby,quantitation of HCPs over a broad dynamic range was enabled to allow monitoring of problematic HCPs or to track changes upon altered bioprocessing conditions.展开更多
Nanopolystyrene was used as a solid support for the covalent immobilization of Candida antarctica lipase B(Cal B) using the photoreactive reagent 1‐fluoro‐2‐nitro‐4‐azido benzene(FNAB) as a cou‐pling reagent. Th...Nanopolystyrene was used as a solid support for the covalent immobilization of Candida antarctica lipase B(Cal B) using the photoreactive reagent 1‐fluoro‐2‐nitro‐4‐azido benzene(FNAB) as a cou‐pling reagent. The obtained derivative was then used as a biocatalyst in a microwave assisted ester‐ification experiment. Factors such as contact time, pH, and enzyme concentration were investigated during immobilization. The hydrolytic activity, thermal, and operational stability of immobi‐lized‐Cal B were determined. The maximum immobilized yield(218 μg/mg support) obtained at pH 6.8 exhibited optimum hydrolytic activity(4.42 × 103 mU p‐nitrophenol/min). The thermal stability of Cal B improved significantly when it was immobilized at pH 10, however, the immobilized yield was very low(93.6 μg/mg support). The immobilized‐Cal B prepared at pH 6.8 and pH 10 retained 50% of its initial activity after incubation periods of 14 and 16 h, respectively, at 60 °C. The opera‐tional stability was investigated for the microwave assisted esterification of oleic acid with metha‐nol. Immobilized‐Cal B retained 50% of its initial activity after 15 batch cycles in the micro‐wave‐assisted esterification. The esterification time was notably reduced under microwave irradia‐tion. The combined use of a biocatalyst and microwave heating is thus an alternative total green synthesis process.展开更多
With the size of the biopharmaceutical market exponentially increasing,there is an aligned growth in the importance of data-rich analyses,not only to assess drug product safety but also to assist drug development driv...With the size of the biopharmaceutical market exponentially increasing,there is an aligned growth in the importance of data-rich analyses,not only to assess drug product safety but also to assist drug development driven by the deeper understanding of structure/function relationships.In monoclonal antibodies,many functions are regulated by N-glycans present in the constant region of the heavy chains and their mechanisms of action are not completely known.The importance of their function focuses analytical research efforts on the development of robust,accurate and fast methods to support drug development and quality control.Released N-glycan analysis is considered as the gold standard for glycosylation characterisation;however,it is not the only method for quantitative analysis of glycoform heterogeneity.In this study,ten different analytical workflows for N-glycan analysis were compared using four monoclonal antibodies.While observing good comparability between the quantitative results generated,it was possible to appreciate the advantages and disadvantages of each technique and to summarise all the observations to guide the choice of the most appropriate analytical workflow according to application and the desired depth of data generated.展开更多
As the roles of glycans in health and disease continue to be unraveled,it is becoming apparent that glycans’immense complexity cannot be ignored.To fully delineate glycan structures,we developed an integrative approa...As the roles of glycans in health and disease continue to be unraveled,it is becoming apparent that glycans’immense complexity cannot be ignored.To fully delineate glycan structures,we developed an integrative approach combining a set of cost-effective,widespread,and easy-to-handle analytical methods.The key feature of our workflow is the exploitation of a removable fluorescent label—exemplified by 9-fluorenylmethyl chloroformate(Fmoc)—to bridge the gap between diverse glycoanalytical methods,especially multiplexed capillary gel electrophoresis with laser-induced fluorescence detection(xCGELIF)and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOFMS).Through the detailed structural analysis of selected,dauntingly complex N-glycans from chicken ovalbumin,horse serum,and bovine transferrin,we illustrate the capabilities of the presented strategy.Moreover,this approach“visualizes”N-glycans that have been difficult to identify thus far—such as the sulfated glycans on human immunoglobulin A—including minute changes in glycan structures,potentially providing useful new targets for biomarker discovery.展开更多
Steam cracking is the dominant technology for producing light olefins,which are believed to be the foundation of the chemical industry.Predictive models of the cracking process can boost production efficiency and prof...Steam cracking is the dominant technology for producing light olefins,which are believed to be the foundation of the chemical industry.Predictive models of the cracking process can boost production efficiency and profit margin.Rapid advancements in machine learning research have recently enabled data-driven solutions to usher in a new era of process modeling.Meanwhile,its practical application to steam cracking is still hindered by the trade-off between prediction accuracy and computational speed.This research presents a framework for data-driven intelligent modeling of the steam cracking process.Industrial data preparation and feature engineering techniques provide computational-ready datasets for the framework,and feedstock similarities are exploited using k-means clustering.We propose LArge-Residuals-Deletion Multivariate Adaptive Regression Spline(LARD-MARS),a modeling approach that explicitly generates output formulas and eliminates potentially outlying instances.The framework is validated further by the presentation of clustering results,the explanation of variable importance,and the testing and comparison of model performance.展开更多
Microbial electrosynthesis(MES) can potentially provide a mean for storing renewable energy surpluses as chemical energy. However, the fluctuating nature of these energy sources may represent a threat to MES, as the m...Microbial electrosynthesis(MES) can potentially provide a mean for storing renewable energy surpluses as chemical energy. However, the fluctuating nature of these energy sources may represent a threat to MES, as the microbial communities that develop on the biocathode rely on the continuous existence of a polarized electrode. This work assesses how MES performance, product generation and microbial community evolution are affected by a long-period(6 weeks) power off(open circuit). Acetogenic and H2-producing bacteria activity recovered after reconnection. However, few days later syntrophic acetate oxidation bacteria and H2-consuming methanogens became dominant, producing CH4 as the main product, via electromethanogenesis and the syntrophic interaction between eubacterial and archaeal communities which consume both the acetic acid and the hydrogen present in the cathode environment. Thus,the system proved to be resilient to a long-term power interruption in terms of electroactivity. At the same time, these results demonstrated that the system could be extensively affected in both end product generation and microbial communities.展开更多
This review offers an overview of the latest developments in metal-covalent organic framework(MCOF)and covalent metal-organic framework(CMOF)materials,whose construction entails a combination of reversible coordinatio...This review offers an overview of the latest developments in metal-covalent organic framework(MCOF)and covalent metal-organic framework(CMOF)materials,whose construction entails a combination of reversible coordination and covalent bonding adapted from metal-organic frameworks(MOFs)and covalent organic frameworks(COFs),respectively.With an emphasis on the MCOF and CMOF structures,this review surveys their building blocks and topologies.Specifically,the frameworks are classified based on the dimensions of their components(building blocks),namely,discrete building blocks and one-dimensional infinite building blocks.For the first category,the materials are further divided into collections of two-and three-dimensional networks based on their topologies.For the second category,the recently emerging MCOFs with woven structures are covered.Finally,the state-of-the-art in MCOF and CMOF chemistry has been laid out for promising avenues in future developments.展开更多
文摘Existing therapies for the treatment of chronic heart failure still have some limitations and there is a pressing need for the development of new therapeutic modalities. The amniotic membrane has been used for the treatment of various diseases, such as conjunctive defects;however, the mechanisms behind its repair functions are still unclear. Regenerative medicine is seeking newer alternatives and among them, biomaterials have emerged in recent years for developing and manipulating molecules, cells, tissues or organs grown in laboratories in order to replace human body parts. Many such materials have been used for this purpose, either synthetically or biologically, in order to provide new medical devices. This review provides a wider view of the regeneration potential of the use of amniotic membrane as a potential biomaterial to facilitate the implementation of new research in surgical procedures. Amniotic membrane appears to be an alternative source of stem cells as well as an excellent biomaterial for cell-based therapeutic applications in engineering heart tissue.
基金funded by the National Council for Scientific and Technological Development,CNPq,grant nos.440343/2022-4 and 306421/2020-8.
文摘The combined inoculation of yeast and lactic acid bacteria(LAB)is a promising approach to enhance microbial metabo-lism and the quality of fermented food products.In this study,we investigated the effects of simultaneous and sequential inoculation of LAB and yeast on coffee fermentation.Fermentations were conducted using single and combined protocols with P.fermentans YC5.2 and P.pentosaceus LPBF07.A temporal analysis utilizing Illumina high-throughput rRNA Gene Sequencing revealed a synergistic interaction between the two microbial groups.This positive synergy led to increased consumption of coffee pulp sugar and the production of metabolites,surpassing the results observed in single cultures and the spontaneous process.Furthermore,the combined inoculation processes demonstrated a more significant role in sup-pressing wild microbiota compared to single cultures.Notably,the sequential process emerged as particularly effective in promoting a more intricate aroma profile and elevated sensorial score,attributed to the formation of distinctive compounds such as benzeneacetaldehyde,1-hexanol,2-heptanol,benzyl alcohol and phenylethyl alcohol.These results suggest that implementing a sequential inoculation process could enable coffee farmers to standardize on-farm processing and produce high-value-added products.
基金Conselho Nacional de Desenvolvimento Científico e Tecnológico,Process 440080/2022-3Luciana Porto de Souza Vandenberghe,409337/2023-4Luciana Porto de Souza Vandenberghe,Coordenação de Aperfeiçoamento de Pessoal de Nível Superior,CAPES-PROEX,Ariane Fátima Murawski de Mello。
文摘Interest in the technology for producing alternative proteins is rapidly increasing,driven by the need to find new ways to produce and consume protein for the global population.This technology involves growing different microorganisms and animal cells under controlled conditions to ensure their viability and efficient growth.The cultivation process takes place in different types of bioreactors,from traditional models to innovative new designs,each offering unique features and capabilities.The most commonly used bioreactors are stirred tank reactors,which are mechanically agitated,and airlift or bubble column bioreactors,which are pneumatically agitated.These bioreactors are often adapted or modified to optimize the production of cultured meat.Essential to the process are microcarriers or scaffolds that support cell adhesion and proliferation.Other bioreactor models,such as hollow fiber and packed bed bioreactors,are also being explored.The trend towards single-use technology is growing due to benefits like easier cleaning and sterilization,and reduced operation times,though it does raise concerns about plastic waste.This review not only describes various bioreactor models but also discusses instrumentation and control systems.It aims to present the main bioreactor models currently in use for cultivated meat production,detailing their features,advantages,disadvantages,and the technological challenges that need to be addressed.
文摘Wet granulation-a unit operation involving mixing polymeric binders with powdered formulations-is well established in the pharmaceutical industry,playing a major role in the manufacturing of oral solid dosage forms and improving the physical properties of granules(size,density,shape factor,etc.)before tableting.The foaming properties of aqueous polymeric binders prove useful for binder delivery within the mixing vessel,with foamed binders leading to enhanced process efficiency(binder distribution,drying time,and temperature)and product quality(heat-sensitive components)during granulation.Given the importance of this stage in producing oral solid dosage forms,understanding the relationship between critical process parameters and critical quality attributes is essential.The process analytical technology(PAT)framework enables process design,analysis,and control and facilitates process development via in-line spectroscopy combined with multivariate data analysis to yield critical product information during the unit operation.Herein,we used in-line NIR spectroscopy to monitor granule size in foam granulations of a pharmaceutical compound.The mean granule diameter was predicted using a partial least squares regression(PLSR)model(with a prediction error of 11.8μm)and combined with a batch statistical process control(BSPC)approach for the temporal monitoring of granule size during three foam granulations.
基金funded by the Natural Sciences and Engineering Research Council of Canada(NSERC)CREATE Program 554777-2021Training in Advanced Biotechnology for Environmental Sustainability(TABES)James and Joanne Love Chair in Environmental Engineering.
文摘The expanding field of alternative proteins represents a transformative approach to addressing global food security and sustainability challenges.Among these,fermentation-derived alternative proteins cultivated from microorganisms such as fungi,bacteria,and algae offer a promising avenue for sustainable protein production.This review explores the selection and utilization of raw materials to produce microbial proteins through fermentation processes.Critical raw materials include agricultural byproducts,industrial waste streams,and specifically designed feedstocks,which not only mitigate environmental footprint but also enhance the economic viability of production systems.The utilization of lignocellulosic biomass and molasses has demonstrated considerable promise,attributed to their abundant and renewable nature.The review underscored the necessity of exploring specific areas to enhance the viability of producing microbial protein from diverse raw materials.These areas include improving pre-treatment strategies to enhance substrate suitability for fermentation,optimizing fermentation processes for increased yield and reduced costs,and developing more resilient microorganisms capable of thriving on varied substrates.These strategies are crucial for advancing the production of alternative proteins through fermentation,in addition to raw material selection,which is vital in the scalability and sustainability of alternative protein production through fermentation,emphasizing the need for continued research and innovation in this field.
基金supported by the Araucária Foundation[NAPI Proteínas Alternativas,grant number 19.810.961-4]the National Council for Scientific and Technological Development-CNPq[grant number 309237/2021-1].
文摘Enzymes play a pivotal role in meat processing and maturation,starting with endogenous enzymes that catalyse a partial proteolysis in natural meat that enhances texture and flavor,to exogenous enzymes that may be incorporated for additional processing.Therefore,a question is raised whether endogenous and exogenous proteases could be used to improve the quality of meat products originated from in vitro cell-based technology.This manuscript aims at presenting the main applications of enzymes in meat processing,starting with conventional meat products and expanding to reported and potential applications in cultured meat technologies.Enzymes are applied since the early stages of cell isolation,multiplication and detaching from supports.The use of endogenous enzymes,such as cathepsins and calpains,has been proposed to improve the palatability of cultivated meat,and the application of transglutaminases has already been reported in scientific papers and patent documents,with the aim of promoting crosslinking reactions among proteins.This feature can be used to modulate the texture of cultivated meat products.Also,their use has been directed to the manufacture of scaffolds and microcarriers for animal cell cultivation.As concluded from the scientific and technological literature survey,it is evident that a substantial area of study remains unexplored regarding the utilization of enzymes in cultivated meat products.
基金funding has been received from National Key Research and Development Program of China with Grant no.2023YFD1201600.
文摘Identifying alternative protein sources is crucial in view of the shortage of protein resources.A new strain Geotrichum can-didum IBB69 was isolated for microbial protein production in this study.The protein yield,biomass,and specific protein production(the ratio of total intracellular and extracellular protein to biomass)of G.candidum IBB69 were 7.6 g/L,18.42 g/L and 60.8%after optimization through shaking flask fermentation,response surface methodology,and 5L scale-up fermen-tation.Compared to flask fermentation,the protein yield and content increased by 83.13%and 91.08%,respectively.The protein product of IBB69 was composed of 18 amino acids with a ratio of 37.51%(EAA to TAA).Transcriptome analysis revealed that,compared with ammonium sulfate,the addition of urea upregulated the expression levels of key genes in the carbon-nitrogen metabolism process of G.candidum IBB69,promoting amino acid synthesis and cell growth.This study tapped a new microbial protein producing strain,and optimized results set the stage for the industrial development and application of G.candidum for sustainable alternative proteins.
文摘在基于核磁共振(NMR)的代谢组学数据分析中,尺度缩放是关键的预处理步骤之一,其主要目的是通过调整数据的方差结构,改善后续的多变量统计分析的结果。从信息熵的角度出发,利用KullbackLeibler(K-L)散度来度量不同实验分组的生物样品的1 H NMR波谱数据的差异程度,并结合单位方差缩放法,提出一种基于K-L散度的尺度缩放方法。该方法先利用单位方差法将数据各变量的标准差调整到同一水平上,再利用K-L散度对各变量进行有监督地加权,增强重要变量、减弱无关变量。由于K-L散度是在概率分布的意义上度量数据间的差异程度,且对于高斯和非高斯分布的数据均适用,因此能更准确地度量不同实验分组样品的1 H NMR波谱数据的差异性,从而更有效地地对谱数据的重要变量进行识别和加权。人群尿液1 H NMR波谱数据的分析结果表明,基于K-L散度的尺度缩放方法能有效抑制噪声变量,同时很好地区分特征变量和非特征变量;提高主成分回归(PCR)模型的判别能力;改善偏最小二乘回归判别分析(PLS-DA)模型的解释能力、预测能力以及对特征代谢物的辨识能力。
基金supported by the National Natural Science Foundation of China,No.30960448the Natural Science Foundation of Jiangxi Province,No.20142BAB205023the Ph.D.Start-up Fund of Natural Science Foundation of Jiangxi Science&Technology Normal University in China,No.3000990122
文摘Methylprednisolone is a commonly used drug for the treatment of spinal cord injury, but high doses of methylprednisolone can increase the incidence of infectious diseases. Methotrexate has anti-inflammatory activity and immunosuppressive effects, and can reduce in- flammation after spinal cord injury. To analyze gene expression changes and the molecular mechanism of methotrexate combined with methylprednisolone in the treatment of spinal cord injury, a rat model of spinal cord contusion was prepared using the PinPointTM preci- sion cortical impactor technique. Rats were injected with methylprednisolone 30 mg/kg 30 minutes after injury, and then subcutaneously injected with 0.3 mg/kg methotrexate 1 day after injury, once a day, for 2 weeks. TreadScan gait analysis found that at 4 and 8 weeks after injury, methotrexate combined with methylprednisolone significantly improved hind limb swing time, stride time, minimum longitudinal deviation, instant speed, footprint area and regularity index. Solexa high-throughput sequencing was used to analyze differential gene ex- pression. Compared with methylprednisolone alone, differential expression of 316 genes was detected in injured spinal cord treated with methotrexate and methylprednisolone. The 275 up-regulated genes were mainly related to nerve recovery, anti-oxidative, anti-inflammatory and anti-apoptotic functions, while 41 down-regulated genes were mainly related to proinflammatory and pro-apoptotic functions. These results indicate that methotrexate combined with methylprednisolone exhibited better effects on inhibiting the activity of inflammatory cytokines and enhancing antioxidant and anti-apoptotic effects and thereby produced stronger neuroprotective effects than methotrexate alone. The 316 differentially expressed genes play an important role in the above processes.
基金financially supported by the grants for the project "Smart Process Inspection" (funding code ZF4184501CR5) from the "Zentrales Innovationsprogramm Mittelstand" (ZIM)
文摘Particles occur in almost all processes in chemical and life sciences. The particle size and shape influence the process performance and product quality, and in turn they are influenced by the flow behavior of the particles during production. Monitoring and controlling such characteristics in multiphase systems to obtain sufficient qualities will greatly facilitate the achievement of reproducible and defined distributions. So far, obtaining this information inline has been challenging, because existing instruments lack measurement precision, being unable to process overlapping signals from different particle phases in highly concentrated multiphase systems. However, recent advances in photo-optics made it possible to monitor such features(particle size distribution(PSD), aspect ratio and particle concentration) with advanced image analysis(IA) in real-time. New analysis workflows as well as single feature extractions from the images using multiple image analysis algorithms allowed the precise real-time measurements of size, shape and concentration of particle collectives even separated from each other in three phase systems. The performances, advantages and drawbacks with other non-photo-optical methods for assessing the particle size distribution are compared and discussed.
基金support from the SFI-NSFC bilateral funding scheme(grant number SFI/17/NSFC/5229)。
文摘Ferrite/carbon composited materials,especially the bio-derived composited materials possessing both environmental friendliness and outstanding microwave absorption performance,attract numerous attentions for solving the"electromagnetic problem"in the Gigahertz frequency range.In this work,we demonstrate a bio-derived ferrite/carbon material by compositing functional carbonized cotton fibers(CCFs)and Fe_(3)O_(4)nanoparticles with optimized microwave-absorption properties.By adjusting the carbonization conditions systematically,the Fe_(3)O_(4)loading contents and the microwave absorption properties can be varied simultaneously-and,indeed,optimized and tuned.The CCFs-Fe_(3)O_(4)composites exhibited a minimum reflection-loss capacity RL(d B)of-56.8 d B at 10.9 GHz with a thickness of 1.67 mm,and its effective absorption bandwidth(RL(d B)<-20 d B)was found to broaden to 7.1 GHz.Electromagnetic characterizations,coupled with microstructure analyses,revealed that the enhancement in microwave absorption was triggered by the different microstructures of CCFs-Fe_(3)O_(4)composites-attributable to the different carbonization processes.These different conditions result in different amounts of Fe_(3)O_(4)attachment sites and lead to the enhancement of dielectric polarization at localized microstructures.The present work of bio-derived ferrite/carbon materials has important implications in understanding structure-performance relationships in dielectric-magnetic materials,and,meanwhile,could well be extended to a microwave-absorber design approach.
基金funding from Thermo Fisher Scientific as part of a funded collaborative agreement with NIBR。
文摘Ensuring the removal of host cell proteins(HCPs) during downstream processing of recombinant proteins such as monoclonal antibodies(m Abs) remains a challenge.Since residual HCPs might affect product stability or safety,constant monitoring is required to demonstrate their removal to be below the regulatory accepted level of 100 ng/mg.The current standard analytical approach for this procedure is based on ELISA;however,this approach only measures the overall HCP content.Therefore,the use of orthogonal methods,such as liquid chromatography-mass spectrometry(LC-MS),has been established,as it facilitates the quantitation of total HCPs as well as the identification and quantitation of the individual HCPs present.In the present study,a workflow for HCP detection and quantitation using an automated magnetic bead-based sample preparation,in combination with a data-independent acquisition(DIA) LC-MS analysis,was established.Employing the same instrumental setup commonly used for peptide mapping analysis of m Abs allows for its quick and easy implementation into pre-existing workflows,avoiding the need for dedicated instrumentation or personnel.Thereby,quantitation of HCPs over a broad dynamic range was enabled to allow monitoring of problematic HCPs or to track changes upon altered bioprocessing conditions.
基金support by the Ministry of Higher Education (MOHE) Malaysia, Ministry ofAgricultural (MOA) and University Teknology Malayisa
文摘Nanopolystyrene was used as a solid support for the covalent immobilization of Candida antarctica lipase B(Cal B) using the photoreactive reagent 1‐fluoro‐2‐nitro‐4‐azido benzene(FNAB) as a cou‐pling reagent. The obtained derivative was then used as a biocatalyst in a microwave assisted ester‐ification experiment. Factors such as contact time, pH, and enzyme concentration were investigated during immobilization. The hydrolytic activity, thermal, and operational stability of immobi‐lized‐Cal B were determined. The maximum immobilized yield(218 μg/mg support) obtained at pH 6.8 exhibited optimum hydrolytic activity(4.42 × 103 mU p‐nitrophenol/min). The thermal stability of Cal B improved significantly when it was immobilized at pH 10, however, the immobilized yield was very low(93.6 μg/mg support). The immobilized‐Cal B prepared at pH 6.8 and pH 10 retained 50% of its initial activity after incubation periods of 14 and 16 h, respectively, at 60 °C. The opera‐tional stability was investigated for the microwave assisted esterification of oleic acid with metha‐nol. Immobilized‐Cal B retained 50% of its initial activity after 15 batch cycles in the micro‐wave‐assisted esterification. The esterification time was notably reduced under microwave irradia‐tion. The combined use of a biocatalyst and microwave heating is thus an alternative total green synthesis process.
文摘With the size of the biopharmaceutical market exponentially increasing,there is an aligned growth in the importance of data-rich analyses,not only to assess drug product safety but also to assist drug development driven by the deeper understanding of structure/function relationships.In monoclonal antibodies,many functions are regulated by N-glycans present in the constant region of the heavy chains and their mechanisms of action are not completely known.The importance of their function focuses analytical research efforts on the development of robust,accurate and fast methods to support drug development and quality control.Released N-glycan analysis is considered as the gold standard for glycosylation characterisation;however,it is not the only method for quantitative analysis of glycoform heterogeneity.In this study,ten different analytical workflows for N-glycan analysis were compared using four monoclonal antibodies.While observing good comparability between the quantitative results generated,it was possible to appreciate the advantages and disadvantages of each technique and to summarise all the observations to guide the choice of the most appropriate analytical workflow according to application and the desired depth of data generated.
基金support from the German Federal Ministry of Education and Research(BMBF)under the project“Die Golgi Glykan Fabrik 2.0”(031A557C for Samanta Cajic and Erdmann Rapp)the European Commission(EC)under the project“HighGlycan”(278535 for RenéHennig and Erdmann Rapp)the Deutsche Forschungsgemeinschaft(DFG,German Research Foundation)under the project“The concert of dolicholbased glycosylation:from molecules to disease models”(FOR2509 for Valerian Grote and Erdmann Rapp).
文摘As the roles of glycans in health and disease continue to be unraveled,it is becoming apparent that glycans’immense complexity cannot be ignored.To fully delineate glycan structures,we developed an integrative approach combining a set of cost-effective,widespread,and easy-to-handle analytical methods.The key feature of our workflow is the exploitation of a removable fluorescent label—exemplified by 9-fluorenylmethyl chloroformate(Fmoc)—to bridge the gap between diverse glycoanalytical methods,especially multiplexed capillary gel electrophoresis with laser-induced fluorescence detection(xCGELIF)and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOFMS).Through the detailed structural analysis of selected,dauntingly complex N-glycans from chicken ovalbumin,horse serum,and bovine transferrin,we illustrate the capabilities of the presented strategy.Moreover,this approach“visualizes”N-glycans that have been difficult to identify thus far—such as the sulfated glycans on human immunoglobulin A—including minute changes in glycan structures,potentially providing useful new targets for biomarker discovery.
基金supported by the National Key Research and Development Program of China(2021 YFB 4000500,2021 YFB 4000501,and 2021 YFB 4000502)。
文摘Steam cracking is the dominant technology for producing light olefins,which are believed to be the foundation of the chemical industry.Predictive models of the cracking process can boost production efficiency and profit margin.Rapid advancements in machine learning research have recently enabled data-driven solutions to usher in a new era of process modeling.Meanwhile,its practical application to steam cracking is still hindered by the trade-off between prediction accuracy and computational speed.This research presents a framework for data-driven intelligent modeling of the steam cracking process.Industrial data preparation and feature engineering techniques provide computational-ready datasets for the framework,and feedstock similarities are exploited using k-means clustering.We propose LArge-Residuals-Deletion Multivariate Adaptive Regression Spline(LARD-MARS),a modeling approach that explicitly generates output formulas and eliminates potentially outlying instances.The framework is validated further by the presentation of clustering results,the explanation of variable importance,and the testing and comparison of model performance.
基金the Spanish“Ministerio de Educación,Cultura y Deporte”for the predoctoral FPU Grant(FPU14/01573)the‘Ministerio de Economía y Competitividad’for the support of project ref:CTQ2015-68925-R(MINECO/FEDER,EU)。
文摘Microbial electrosynthesis(MES) can potentially provide a mean for storing renewable energy surpluses as chemical energy. However, the fluctuating nature of these energy sources may represent a threat to MES, as the microbial communities that develop on the biocathode rely on the continuous existence of a polarized electrode. This work assesses how MES performance, product generation and microbial community evolution are affected by a long-period(6 weeks) power off(open circuit). Acetogenic and H2-producing bacteria activity recovered after reconnection. However, few days later syntrophic acetate oxidation bacteria and H2-consuming methanogens became dominant, producing CH4 as the main product, via electromethanogenesis and the syntrophic interaction between eubacterial and archaeal communities which consume both the acetic acid and the hydrogen present in the cathode environment. Thus,the system proved to be resilient to a long-term power interruption in terms of electroactivity. At the same time, these results demonstrated that the system could be extensively affected in both end product generation and microbial communities.
基金financially supported by the National Natural Science Foundation of China(No.22176135)the Fundamental Research Funds for the Central Universities in China(No.YJ201976)。
文摘This review offers an overview of the latest developments in metal-covalent organic framework(MCOF)and covalent metal-organic framework(CMOF)materials,whose construction entails a combination of reversible coordination and covalent bonding adapted from metal-organic frameworks(MOFs)and covalent organic frameworks(COFs),respectively.With an emphasis on the MCOF and CMOF structures,this review surveys their building blocks and topologies.Specifically,the frameworks are classified based on the dimensions of their components(building blocks),namely,discrete building blocks and one-dimensional infinite building blocks.For the first category,the materials are further divided into collections of two-and three-dimensional networks based on their topologies.For the second category,the recently emerging MCOFs with woven structures are covered.Finally,the state-of-the-art in MCOF and CMOF chemistry has been laid out for promising avenues in future developments.
