Production Platform Research Articles

First-class - biosynthesis of 6-MSA and bostrycoidin type I polyketides in Yarrowia lipolytica.

Fungal polyketides are a large group of secondary metabolites, valuable due to their diverse spectrum of pharmacological activities. Polyketide biosynthesis in filamentous fungi presents some challenges: small yield and low-purity titers. To tackle these issues, we switched to the yeast Yarrowia lipolytica, an easily cultivable heterologous host. As an oleaginous yeast, Y. lipolytica displays a high flux of acetyl- and malonyl-CoA precursors used in lipid synthesis. Likewise, acetyl- and malonyl-CoA are the building blocks of many natural polyketides, and we explored the possibility of redirecting this flux toward polyketide production. Despite its promising prospect, Y. lipolytica has so far only been used for heterologous expression of simple type III polyketide synthases (PKSs) from plants. Therefore, we decided to evaluate the potential of Y. lipolytica by targeting the more complex fungal polyketides synthesized by type I PKSs. We employed a CRISPR-Cas9-mediated genome editing method to achieve markerless gene integration of the genes responsible for bostrycoidin biosynthesis in Fusarium solani (fsr1, fsr2, and fsr3) and 6-methylsalicylic acid (6-MSA) biosynthesis in Aspergillus hancockii (6MSAS). Moreover, we attempted titer optimization through metabolic engineering by overexpressing two enzymes, TGL4 and AOX2, involved in lipid β-oxidation, but we did not observe an effect on polyketide production. With maximum titers of 403 mg/L 6-MSA and 35 mg/L bostrycoidin, the latter being substantially higher than our previous results in Saccharomyces cerevisiae (2.2 mg/L), this work demonstrates the potential of Y. lipolytica as a platform for heterologous production of complex fungal polyketides.

Abstract 2760: Novel GPCR recombinant monoclonal antibodies for cancer biology research

Abstract Recombinant technology is becoming the preferred production mechanism for creation of antibody reagents used in academic and clinical cancer biology research. Recombinant antibodies (rAbs) are superior to standard polyclonal and hybridoma-generated monoclonal reagents as they offer reproducible performance and consistent, scalable supply. A major focus of antibody manufacturers is to leverage rAb production strategies to generate antibodies against challenging targets for which there are few, if any, reliable antibodies. G-protein coupled receptors (GPCRs) are encoded by more than 820 human genes and represent the largest membrane protein superfamily in the genome. These seven-pass transmembrane domain receptors are involved in a multitude of physiological and pathophysiological processes related to various chronic diseases, including an array of malignancies. Perhaps 30-40% of all therapeutics are directed against GPCR activity, and ~350 of the non-olfactory GPCRs are thought to be potential candidates for drug development. However, for a slew of technical reasons, the development and validation of high-quality antibodies against GPCRs has proven difficult. In an effort to facilitate further research into GPCRs, GeneTex is combining its recombinant antibody production platform with enhanced validation techniques to make specific monoclonal antibodies against GPCRs with direct relevance to cancer and other diseases. GeneTex’s rAb production protocol employs a multi-parameter fluorescence-activated single cell sorting (FACS)-based methodology to identify and select antigen-specific IgG+ memory B cells from an immunized rabbit (Starkie et al., 2016). The heavy and light chain variable region genes from single cells are amplified, cloned, and co-expressed in mammalian cells to generate a monoclonal IgG. Application-specific testing is performed in conjunction with knockout/knockdown (KO/KD), differential expression comparison in cells and tissues, cell fraction enrichment (i.e., membrane extracts versus whole cell extracts), and other methodologies. Antibodies directed against extracellular GPCR domains are validated for specificity using GPCR arrays (CDI Labs, Baltimore, MD). This workflow has resulted in the production of specific antibodies for a number of GPCRs studied by cancer biologists. These include novel recombinant monoclonal antibodies to the dopamine D2 receptor (DRD2), retinoic acid-induced protein 3 (RAI3), CXC motif chemokine receptor 2 (CXCR2), CXCR4, and CXCR7. All of these antibodies were validated for at least two applications, while the specificity of three (i.e., RAI3, CXCR4, and CXCR7) is supported by KO/KD data. GPCR array data argues for the specificity of the other two antibodies (i.e., DRD2 and CXCR2). This same production/validation approach will be applied as GeneTex targets the remaining GPCRs. Citation Format: Alexander Ball, Yung Lin Hsieh, Chun Kai Huang, Sega Huang, Shang Ru Wu, Yue Yun Chang, Jiming Liu, Po Shin Peng, Chia Yi Lin. Novel GPCR recombinant monoclonal antibodies for cancer biology research [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2760.

Abstract 6773: Tissue engineered colorectal cancer microspheres for drug screening

Abstract Three-dimensional (3D) disease models have garnered widespread interest for use in later stages of the drug discovery process, such as preclinical efficacy and toxicology studies, due to their pathophysiologically relevant properties. However, there is a need and opportunity for 3D cancer models to be used earlier in the drug screening process. To meet this need, the 3D models must strike a balance between throughput, which includes scalability and uniformity, and physiological relevance, such as the ability to modulate key attribute of the tumor microenvironment. Here we report the creation of 3D colorectal cancer (CRC) tissue models, referred to as VivoSpheres, and demonstrated their relevance to cancer drug screening. The VivoSphere production platform couples tissue engineering toolkits with microfluidics, enabling the scalable production of engineered cancer microspheres. The model supports the long-term maintenance of the cancer cell phenotype. In a preliminary study, we were able to generate more physiologically relevant drug responses. We formed CRC VivoSpheres by encapsulating HT-29 CRC cells within poly(ethylene glycol)-fibrinogen hydrogel microspheres using our previously developed microfluidic platform. CRC VivoSpheres were rapidly produced with high cell densities (20 × 106 cells/ml) and high uniformity on day 0 with a coefficient of variation (COV) < 7%. This high uniformity was maintained for 15 days (COV ≤ 10%), which is critical for long-term dose studies. The cells maintained high viability and showed high proliferative capability with a significant increase in colony size and expression of Ki67 up to day 29. The encapsulated cells maintained the CRC phenotype over time with the expression of CD44 (cancer stem cell marker) and CK20 (CRC marker). After establishing shipping conditions that maintained cell viability for remote use, the HT-29 VivoSpheres were shipped to the oncology team at Southern Research for drug testing. The CRC VivoSpheres were treated with DMSO, GANT61, and SRI-38832, the latter two of which are GLI1 inhibitors. Phase contrast images and western blot were used to assess the response of CRC VivoSpheres to the treatments. Oncogenic GLI1 transcription activity and NBS1 overexpression have been found to contribute to chemotherapeutic resistance, negating the anti-tumor effects of 5-fluorouracil. While 2D cultured HT-29s responded to treatment with GANT61, HT-29 VivoSpheres continued to express NBS1 following GANT1 treatment, but downregulated NBS1 in response to the GLI1 inhibitor SRI-38832, which is the same response Southern Research has seen in in vivo tumor models. In conclusion, we have developed tissue-engineered 3D CRC models that hold promise for use in drug screening. These models have demonstrated an initial capability to reproduce the CRC phenotype and mimic in vivo drug response. Citation Format: Yuan Tian, Mohammadjafar Hashemi, Benjamin Anbiah, Ruowen Zhang, Iman Hassani, Rebecca Boohaker, Elizabeth Lipke. Tissue engineered colorectal cancer microspheres for drug screening [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6773.

Abstract 6756: Our RNA production platform and the associated tools facilitate the development of RNA-based vaccine

Abstract mRNA vaccines have shown their full potential in the Covid-19 pandemic. The application of these technologies for the treatment of cancers represents significant hope, but clinical effectiveness remains to be demonstrated. For this purpose, researchers require high quality production of vaccine mRNA and accurate and cost-saving test tools that are adapted to oncology research. Since 2021, Tebubio has developed a miniscale RNA production platform for providing RNAs (from 100 µg to 1 mg) to researchers around the world. This service is completed by offers on RNA delivery solutions using lipid nanoparticles (LNPs) and cellular tests with biomarker analysis. Through an ingenious design, our DNA matrix for mRNA is easily transferable for GMP production for clinical studies. Recently, Tebubio has developed DNA matrices for circular RNA production. Indeed, circular RNA has many advantages by being less immunogenic and more stable than linear RNA. We compared the quality and the quantity of CAP1 capped linear RNA and circular RNA produced from our DNA templates. We also assessed the stability of both RNAs expressing GFP and the typical tumor antigen, p53, and their relative expression into antigen-presenting cells (APCs). Finally, we quantified the response of the APCs expressing p53 by using our biomarker multiplex analysis system. This new type of RNA and its efficiency testing in APCs will constitute a new addition to our pipeline of RNA production and testing for preclinical studies. Citation Format: Célia Bosso-Lefèvre, Erica Cirri, Eric Mennesson, Dimitri Szymczak, Flavien Carpentier, Alexandra Foucher, Alzbeta Komarkova, Maxime Rochet, Armelle Vindrios, Romain Goulay. Our RNA production platform and the associated tools facilitate the development of RNA-based vaccine [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6756.

Real-time prediction of mooring tension for semi-submersible platforms

The harsh operating environment of deepwater semi-submersible production platforms poses severe challenges to the safe operation and maintenance of their production processes. Using artificial intelligence technology to improve the operation and maintenance level of large-scale offshore engineering equipment has become an inevitable trend. Accurate mooring tension prediction for semi-submersible production platforms can effectively identify potential damage and ensure the platform's safe and efficient production operation. Compared with traditional cyclic neural networks, transformer neural networks have faster calculation speeds, better generalization abilities and scalability, and better abilities to capture long-distance dependencies. In this study, the first deepwater semi-submersible production platform in the South China Sea, “DeepSea One,” was selected as the research object, and an improved novel transformer neural network architecture was proposed for the mooring tension prediction of semi-submersible production platforms using actual long-term mooring tension data obtained during the platform's production process. The proposed method was compared with the commonly used long short-term memory neural network model, and the comparison proved the effectiveness of the proposed method in predicting the dynamic tension of semi-submersible production platform mooring. In addition, the proposed method is also applicable for the prediction of other time series data in the field of shipbuilding and offshore engineering, which is of certain value for generalization.

Process design and economic evaluation for methane and electricity production from Agave bagasse using different pretreatments

Agave bagasse is an agricultural residue that has the potential to be used as feedstock biorefineries. However, its complex structure requires a pretreatment step to facilitate the accessibility to carbohydrates for its subsequent conversion. In this study, the capital and operative expenses (CAPEX and OPEX) were simulated for industrial plants processing 500 ton/d of Agave bagasse using different pretreatment platforms for methane production and power generation on experimental data. The scenarios consisted of i) biological, ii) enzymatic, iii) dilute acid, and iv) steam explosion pretreatments and a case-based scenario without pretreatment. All these scenarios included milling, pretreatment, anaerobic digestion, and cogeneration stages. The results showed that those pretreatments carried out under soft conditions (biological and enzymatic) decreased between 2.1 and 2.8 times CAPEX compared to the highest value obtained in the diluted acid and case-based scenarios. The steam explosion, a pretreatment carried out under severe conditions, was less effective reducing CAPEX by only 1.2 times compared to the highest value. Furthermore, compared to the case-base scenario, only the biological pretreatment reduced the OPEX by a factor of 2, whereas the other pretreatments resulted in OPEX that were 2.0 to 3.5 times higher. Among the utilities (cooling water, steam generation, and energy) the steam generation contributed 47% to 76% of the total OPEX. The energy produced (kW⋅h/yr) was approximately twice the energy consumed for all scenarios. A sensitivity analysis demonstrated the robustness of the platforms since fluctuations in the prices of feedstock and raw materials did not impact the total production cost of methane, except for biological and enzymatic pretreatments. For all platforms, the solid loading was the process parameter with the highest impact on the production cost of methane. Overall, the study showed that biological and enzymatic pretreatments had the best economic performance. However, low substrate conversion and the high cost of enzymes and equipment, respectively, remained the main challenges to address in future research.

Efficient production of butyl butyrate from mannitol by engineered Clostridium tyrobutyricum

Butyl butyrate has a wide range of applications as flavors, solvents, and biofuels. Green production of butyl butyrate by microorganism fermentation in an industry level attracts great attention on the basis of bio-refinery. In this study, a dark horse Clostridium tyrobutyricum was employed for de novo production of butyl butyrate. Herein, reinforcement of EMP and butyl butyrate fermentation pathways (e.g., pfkA, pykA, crt-hbd-bcd/etfAB, adhE2 and VAAT) and regulation of carbon distribution towards butyl butyrate (e.g., CAT1 inactivation or overexpression of NADPH-dependent hbd1) were conducted in C. tyrobutyricum (MUV3) to enhance butyl butyrate production. The outstanding strain MV12 generated 1.8-fold butyl butyrate production compared to the initial strain MUV3. Subsequently, organic nitrogen source concentration was optimized, 21.1 g/L and 30.1 g/L butyl butyrate were obtained in 37 °C and 25 °C, respectively. Ultimately, fed-batch with two-stage temperature shift fermentation was performed for high titer and yield of butyl butyrate production (142.6 g/L titer and 0.28 mol/mol yield) from mannitol. Hence this study presented a microbial fermentation platform for butyl butyrate or other value-added chemicals production from renewable resources.

Media Guidelines, Rules and Regulations and Perceived Customer Worth in Vision Group Media in Uganda

This research focused on the media guidelines, rules and importance of customers in Vision Group Media in Uganda. The study design was a case study technique. It used both qualitative and quantitative methods to gain in-depth understanding of performance of the media sector. The population of the study consisted of 176 respondents drowned from Vision Group mangers, supervisors and key stake holders like distributors, readers and listeners. This category was selected because they are believed to be knowledgeable about the subject matter and the company.The results of the research show how there was a favorable correlation between media rules and importance of customers. The results indicate how the company complied with media regulation practices as shown by (74.51%) response. The results on the level of clients’ commitment on different product platforms offered by VG indicates that (74.48%) of the respondents are committed. The key indicators are high on Vision Group rates aligning with the industry standards (50.8%); effective management of client’s feedback (51.6%) and Vision Group print products meeting for audience needs across all categories (50%). The findings from indicate a significant positive correlation between media regulations and customer perceived value (r =0.24). The study recommends that; Vision should continue developing flexible media mix aimed at reaching consumers across the country as each market niche has varying needs, therefore niche marketing tactics should be developed to access these markets. Vision Group and other media houses in general should focus attention on telling brand stories in snippets, with different narrative styles and a balance of information and entertainment, thus allowing consumers to find different pieces in different places. Keywords: Media, Vision group, Customer worth, Media regulations

Debottlenecking the L-DOPA 4,5-dioxygenase step with enhanced tyrosine supply boosts betalain production in Nicotiana benthamiana.

Synthetic biology provides emerging tools to produce valuable compounds in plant hosts as sustainable chemical production platforms. However, little is known about how supply and utilization of precursors is coordinated at the interface of plant primary and specialized metabolism, limiting our ability to efficiently produce high levels of target specialized metabolites in plants. L-Tyrosine is an aromatic amino acid precursor of diverse plant natural products including betalain pigments, which are used as the major natural food red colorants and more recently a visual marker for plant transformation. Here, we studied the impact of enhanced L-tyrosine supply on the production of betalain pigments by expressing arogenate dehydrogenase (TyrA) from table beet (Beta vulgaris, BvTyrAα), which has relaxed feedback inhibition by L-tyrosine. Unexpectedly, betalain levels were reduced when BvTyrAα was coexpressed with the betalain pathway genes in Nicotiana benthamiana leaves; L-tyrosine and 3,4-dihydroxy-L-phenylalanine (L-DOPA) levels were drastically elevated but not efficiently converted to betalains. An additional expression of L-DOPA 4,5-dioxygenase (DODA), but not CYP76AD1 or cyclo-DOPA 5-O-glucosyltransferase, together with BvTyrAα and the betalain pathway, drastically enhanced betalain production, indicating that DODA is a major rate-limiting step of betalain biosynthesis in this system. Learning from this initial test and further debottlenecking the DODA step maximized betalain yield to an equivalent or higher level than that in table beet. Our data suggest that balancing between enhanced supply ("push") and effective utilization ("pull") of precursor by alleviating a bottleneck step is critical in successful plant synthetic biology to produce high levels of target compounds.

Unveiling the repressive mechanism of a PPS-like regulator (PspR) in polyhydroxyalkanoates biosynthesis network.

Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) is a type of polyhydroxyalkanoates (PHA) that exhibits numerous outstanding properties and is naturally synthesized and elaborately regulated in various microorganisms. However, the regulatory mechanism involving the specific regulator PhaR in Haloferax mediterranei, a major PHBV production model among Haloarchaea, is not well understood. In our previous study, we showed that deletion of the phosphoenolpyruvate (PEP) synthetase-like (pps-like) gene activates the cryptic phaC genes in H. mediterranei, resulting in enhanced PHBV accumulation. In this study, we demonstrated the specific function of the PPS-like protein as a negative regulator of phaR gene expression and PHBV synthesis. Chromatin immunoprecipitation (ChIP), in situ fluorescence reporting system, and in vitro electrophoretic mobility shift assay (EMSA) showed that the PPS-like protein can bind to the promoter region of phaRP. Computational modeling revealed a high structural similarity between the rifampin phosphotransferase (RPH) protein and the PPS-like protein, which has a conserved ATP-binding domain, a His domain, and a predicted DNA-binding domain. Key residues within this unique DNA-binding domain were subsequently validated through point mutation and functional evaluations. Based on these findings, we concluded that PPS-like protein, which we now renamed as PspR, has evolved into a repressor capable of regulating the key regulator PhaR, and thereby modulating PHBV synthesis. This regulatory network (PspR-PhaR) for PHA biosynthesis is likely widespread among haloarchaea, providing a novel approach to manipulate haloarchaea as a production platform for high-yielding PHA. KEY POINTS: • The repressive mechanism of a novel inhibitor PspR in the PHBV biosynthesis was demonstrated • PspR is widespread among the PHA accumulating haloarchaea • It is the first report of functional conversion from an enzyme to a trans-acting regulator in haloarchaea.

Antigen Presenting Cell Mimetic Lipid Nanoparticles for Rapid mRNA CAR T Cell Cancer Immunotherapy.

Chimeric antigen receptor (CAR) T cell therapy has achieved remarkable clinical success in the treatment of hematological malignancies. However, producing these bespoke cancer-killing cells is a complicated ex vivo process involving leukapheresis, artificial T cell activation, and CAR construct introduction. The activation step requires the engagement of CD3/TCR and CD28 and is vital for T cell transfection and differentiation. Though antigen-presenting cells (APCs) facilitate activation in vivo, ex vivo activation relies on antibodies against CD3 and CD28 conjugated to magnetic beads. While effective, this artificial activation adds to the complexity of CAR T cell production as the beads must be removed prior to clinical implementation. To overcome this challenge, this work develops activating lipid nanoparticles (aLNPs) that mimic APCs to combine the activation of magnetic beads and the transfection capabilities of LNPs. It is shown that aLNPs enable one-step activation and transfection of primary human T cells with the resulting mRNA CAR T cells reducing tumor burden in a murine xenograft model, validating aLNPs as a promising platform for the rapid production of mRNA CAR T cells.

Biosynthetic pathway redesign in non-conventional yeast for enhanced production of cembratriene-ol

Cembratriene-ol (CBT-ol), a plant-derived macrocyclic diterpene with notable insecticidal activity, has attracted considerable attention with respect to the development of sustainable and green biopesticides. Currently, CBT-ol production is limited by an inefficient and costly plant extraction strategy. Herein, CBT-ol production was enhanced by redesigning the CBT-ol biosynthetic pathway in Candida tropicalis, with subsequent truncation of CBT-ol synthase further increasing CBT-ol production. Moreover, bottlenecks in the CBT-ol biosynthetic pathway were eliminated by adjusting the gene dosage of the rate-limiting enzymes. Ultimately, the resulting strain C. tropicalis CPPt-03D produced 129.17 mg/L CBT-ol in shaking flasks (a 144-fold increase relative to that of the initial strain C01-CD) with CBT-ol production reaching 1,425.76 mg/L in a 5-L bioreactor, representing the highest CBT-ol titer reported to date. These findings provide a green process and promising platform for the industrial production of CBT-ol and lays the foundation for organic farming.

Production and bioprocessing of Taxol from Aspergillus niger, an endophyte of Encephalartos whitelockii, with a plausible biosynthetic stability: antiproliferative activity and cell cycle analysis

The biosynthetic potency of Taxol by fungi raises their prospective to be a platform for commercial production of Taxol, nevertheless, the attenuation of its productivity with the fungal storage, is the challenge. Thus, screening for a novel fungal isolate inhabiting ethnopharmacological plants, with a plausible metabolic stability for Taxol production could be one of the most affordable approaches. Aspergillus niger OR414905.1, an endophyte of Encephalartos whitelockii, had the highest Taxol productivity (173.9 μg/L). The chemical identity of the purified Taxol was confirmed by HPLC, FTIR, and LC–MS/MS analyses, exhibiting the same molecular mass (854.5 m/z) and molecular fragmentation pattern of the authentic Taxol. The purified Taxol exhibited a potent antiproliferative activity against HepG-2, MCF-7 and Caco-2, with IC50 values 0.011, 0.016, and 0.067 μM, respectively, in addition to a significant activity against A. flavus, as a model of human fungal pathogen. The purified Taxol displayed a significant effect against the cellular migration of HepG-2 and MCF-7 cells, by ~ 52–59% after 72 h, compared to the control, confirming its interference with the cellular matrix formation. Furthermore, the purified Taxol exhibited a significant ability to prompt apoptosis in MCF-7 cells, by about 11-fold compared to control cells, suppressing their division at G2/M phase. Taxol productivity by A. niger has been optimized by the response surface methodology with Plackett–Burman Design and Central Composite Design, resulting in a remarkable ~ 1.6-fold increase (279.8 μg/L), over the control. The biological half-life time of Taxol productivity by A. niger was ~ 6 months of preservation at 4 ℃, however, the Taxol yield by A. niger was partially restored in response to ethyl acetate extracts of E. whitelockii, ensuring the presence of plant-derived signals that triggers the cryptic Taxol encoding genes.

Analysis of cell growth, photosynthetic behavior and the fatty acid profile in Tetraselmis subcordiformis under different lighting scenarios

Tetraselmis has been investigated as a potential source of lipids. This microalga possesses good growth characteristics and can be used to develop viable platforms for fatty acid production. This research aims to evaluate the effect of high photon flux density with light-dark cycles and light wavelength on biomass production and fatty acid profile in Tetraselmis subcordiformis. A low light control and treatments with high photon flux density with different light-dark cycles (24:0 h, 12:12 h, 1:1 h, and 15:15 min) and different light wavelength (white, green, red, and blue) were evaluated to determine cell concentration, nutrient consumption, chlorophyll content, photosynthetic yields, lipid content, and fatty acid profile. Significant differences were found in all variables, except for phosphate consumption. High photon flux density promotes cell growth with T. subcordiformis reaching biomass productivities of 0.10 g L-1 day-1 when continuous white light is used. However, no differences were observed in biomass productivities and lipid content for all high photon flux density treatments. On the other hand, red light resulted in higher cell growth, with a productivity of 0.12 g L-1 day-1, and the highest lipid content was achieved under white light. There was a significant effect on the fatty acid profile under different light conditions, with palmitic acid, oleic acid, and eicosapentaenoic acid being the most abundant. This study demonstrated that cellular growth and fatty acid profiles in T. subcordiformis can be influenced by different lighting schemes in the cultivation.

A Sustainable Product Multi-Platform Planning Model for Assembly and Disassembly Process

Abstract The development of a product platform is an effective strategy to respond to dynamic market demands, decrease lead time, and delay product differentiation. However, the traditional product platform configuration method cannot satisfy the sustainability requirements for modern products. To solve this problem, a sustainable product multi-platform (SPMP) model for assembly/disassembly technology is proposed in this article. The proposed SPMP model measures the energy consumption of module instances during the installation based on the platform-based assembly index (PAI) and platform-based disassembly index (PDI), and provides a multi-platform solution for the assembly of the product family. To demonstrate the effectiveness of the proposed method, two product family cases are discussed. A simplified case shows that multi-objective particle swarm optimization (MOPSO) algorithm has a stronger optimization ability than the linear programing method in reducing the product processing cost. The hair dryer family case demonstrates that the proposed method reduces the energy consumption during assembly by linking sustainability to the product design.

Activating the leukocytes by short synthetic peptides in vitro as a stage in the creation of treatment-prophylactic vaccines against COVID-19

Experience with pandemics strongly suggests that vaccination of the population in all categories should be a national priority. The choice of a vaccine production platform should be made in such a way as to achieve an optimal effect at the lowest possible cost. A peptide vaccine or a protein platform vaccine could serve these purposes. Oral and intranasal vaccines are also attractive due to the ease of administration to different population groups, and the resulting immunity is not inferior to that of intramuscularly administered vaccines.In this work, synthetic peptides representing the fragments of the surface protein SARS-CoV-2 were investigated. The peptides were prepared by classical peptide synthesis, with peptide No. 1 (Lys-Ile-Ala-Asp-Tyr-Asn-Tyr-Lys-Leu) being immunodominant for the HLA-A02:01 phenotype with a low calculated concentration of half-maximum inhibition. Peptide No. 2 (Val-Arg-Gln-Ala-Pro-Asn-Gly-Gln-Thr) was chosen as control and is not immunodominant for the HLA-A02:01 phenotype, with a high estimated concentration of half-maximum inhibition (IC50).80 persons were questionnaired and 78 volunteers were examined. Cellular immunity parameters were analyzed using a Cytomics FC 500 flow cytometer and gamma interferon (IFN-γ) was determined by ELISA. The results were processed using Statistica 10 software. As a result, a new method was tested to evaluate the activation of blood leukocytes by synthetic peptides. Regardless of the HLA-A phenotype of the study subjects, the peptides were able to bind to leukocytes, indicating a universal response to foreign peptides, especially to innate immune cells. Peptide No. 2 with high calculated IC50, compared to peptide No. 1 with low calculated IC50, showed significantly higher binding to lymphocytes and monocytes and activation of basophils. The peptides used in this work showed that they interact with leukocytes, activating them through the secretion of IFN-γ. Thus, our work demonstrates an approach to creating a peptide vaccine in the in vitro research phase, as well as to studying the antiviral response by the IFN-γ growth in response to the peptides.

Empowering Rural Revitalization: Unleashing the Potential of E-commerce for Sustainable Industrial Integration

AbstractThe rural revitalization strategy has spotlighted the agricultural industry as a pivotal avenue for national development. E-commerce, as a novel business paradigm, presents both opportunities and challenges to this sector’s evolution. This study endeavors to navigate the trajectory of e-commerce within rural revitalization by amalgamating literature review and case analyses. It seeks to uncover the sales channels offered by e-commerce platforms for agricultural products, subsequently elevating the value proposition of these products and propelling rural economic growth. Findings underscore the pivotal role of e-commerce in rural revitalization. Leveraging e-commerce platforms to bolster the agricultural industry presents innovative avenues and strategies. Nonetheless, challenges loom over these platforms, encompassing issues related to agricultural product quality, logistics, and distribution. Thus, a strategic imperative emerges to fortify logistics infrastructure, enhance agricultural product quality, institute robust personnel training initiatives, and catalyze e-commerce advancements to foster the revitalization and expansion of the agricultural sector.

Slug-flow synthesis of NCMA: Effect of substitution of cobalt with aluminum on the electrochemical performance of Ni-rich cathode for lithium-ion battery

Nickle-rich Li[Ni1-x-yCoxMny]O2 (x, y ≤ 0.1) (NCM) layered materials are known as promising cathode materials for next-generation lithium-ion batteries and electric vehicles owing to their high-reversible capacity and operating voltage of up to 3.6 vs Li/Li+. However, issues, such as irreversible phase transition, cation mixing, microcrack formation, thermal and structural stability of the material prevent its widespread adoption. Although, cation doping is a well-known technique to enhance the electrochemical performance of the NCM-based cathode material, the performance of the material is very sensitive to the doping amount. In this study, three Al-doped quaternary Ni-rich cathode materials Li[Ni0.85Co(0.1-x)Mn0.05Alx]O2 (where, x = 0–0.04) (NCMA) are synthesized through three-phase slug-flow based continuous manufacturing process followed by high temperature calcination to study the effect of Al-doping on the performance of the cathode material while reducing Co. The slug flow-based production platform has several advantages, like particle size uniformity, high production rate, and homogeneity in elemental distribution. It is found that with an increase in Al content, the specific capacity decreases but the cyclic stability and rate capability increases. Optimum Al-doping not only compensates for the adverse effect of low Co by decreasing the extent of cation mixing, but it also minimizes the electrode polarization and cracking of the particles.

A model-based performance analysis of integrated chromatography-ultrafilter separation systems for monoclonal antibody (mAb) manufacturing

Robust and accurate modelling of bioprocess systems offers the opportunity to visualise strategies for rapid-scale up of production platforms from benchtop to industrial scale, but few studies investigate how mechanistic and hybrid models can be used to design integrated bio-manufacturing. This is especially true for downstream purification technologies used to separate mAbs from culture by-products, e.g. host cell proteins. Polishing chromatography (cation-exchange mode) is commonly used to remove these by-products, but optimal bind and elution operation of these columns is complex. Ultrafiltration is typically applied in conjunction with diafiltration post-polishing chromatography in order to obtain suitable final product titers and excipient concentrations. This study presents a combination of dynamic models to probe batch polishing chromatography and ultrafiltration operating strategies achieving industrial specifications for recovery yield and mAb titer. A series of elution and filtration strategies are implemented, resulting in a total of 40 flowsheets explored. A steric mass action model with 7 pH-dependent parameters is employed to simulate scale-up polishing chromatography column behaviour. Moreover, a published gel layer ultrafiltration model based on Darcy’s law is implemented to purify the outlet of the polishing chromatography column. Strategies that are successful in achieving the target 90% recovery yield and 100 g L−1 concentration specifications are compared via a comprehensive analysis, to deduce the operational protocols which offer minimised salt consumption and pressure drop whilst respecting design criteria.

Hybrid Energy Holds Promise for Artificial Lift for Offshore Brazilian Fields

_ This article, written by JPT Technology Editor Chris Carpenter, contains highlights of paper OTC 32671, “Technical Evaluation of the Use of Hybrid Energy (Solar and Offshore Wind) To Supply Artificial Lift Pumps on an Oil Platform on the Equatorial Margin,” by W.K. Silva, Ana L.R. Cunha, SPE, and Ana C. Alves, Federal University of Pará, et al. The paper has not been peer reviewed. _ The complete paper investigates the relevance of wind and solar energy for the supply of artificial lift systems on offshore platforms in the Brazilian equatorial region. The scope of the authors’ study encompassed a detailed technical and environmental assessment, with an emphasis on the integration of a 10-MW floating solar photovoltaic (FSPV) power system to meet demand for 10 pumps. The results obtained highlight the high viability of the system in meeting the demand of these pumps and ensuring an adequate and sustainable energy supply. Area of Study The equatorial margin is an area in northern Brazil covering part of the sedimentary basins of the equatorial Atlantic. The region is home to important sedimentary basins such as the Amazon Basin, the Pará-Maranhão Basin, the Barreirinhas Basin, the Potiguar Basin, and the Ceará Basin, all known for their hydrocarbon reserves. Offshore oil-exploration activities in the equatorial margin involve the installation of drilling and production platforms that include artificial lift systems. To power these systems, a reliable and continuous power source is required. The use of green energies such as wind and solar can be advantageous in meeting this purpose. For the present study, the coastal city of Salinópolis in the state of Pará was selected because of its plentiful meteorological data and its location in the Pará-Maranhão Basin region. Methodology The methodology of the work consists of the evaluation of the energy required for artificial lifting systems with 10 pumps, using the nine steps developed by Baker Hughes’ Artificial Lift Research and Technology Center for the calculation of the power required to drive the pumps. Additionally, the study uses the System Advisor Model (SAM), November 2022 version, developed by the National Renewable Energy Laboratory to simulate the FSPV. Information related to wind potential was obtained through the measuring stations of the National Institute of Meteorology (INMET).