Enhancement Of Production Research Articles

Abstract 1027: Immunohistochemistry (IHC) to assess tumor cell-associated OX40 expression in lymphoma PDXs

Abstract Background. Tumor necrosis factor receptor superfamily member 4, or OX40, is an immunostimulatory receptor usually expressed in activated T-cells. The OX40-OX40L (ligand of OX40) interaction plays a crucial role in the enhancement of T-cell proliferation, survival, and cytokine production, including those in tumor microenvironment (TME) leading to increased anti-tumor immunity1. OX40 agonist antibodies have been widely tested for immunotherapy for solid tumors2,3. Interestingly, in the treatment of lymphoma, the situation may become more complicate, where OX40 could also be found to be expressed on lymphoma cells, in addition to TME. This phenomenon might or might not impact OX40 agonist therapy. This report aims at developing an immunohistochemistry (IHC) assay to assess tumor-associated OX40 by utilizing morphology approach to understand OX40 expression in lymphoma cells, as opposed on tumor-infiltrate T-lymphocytes (TILs) present in TME. FFPE samples from patient-derived xenografts (PDXs), recapitulating the main pathological features of the original tumors, but exclusive of human TILs, were used as the first stage of the assay development, avoiding TIL interference prior to human sample testing. Methods. Ninety leukemia and lymphoma PDXs are comprehensively annotated, including RNAseq and histopathology (HuBaseTM: https://hubase.crownbio.com). IHC staining of PDX FFPE slides or TMA (tissue microarray) of various cancers was performed using a commercial OX40 antibody (CST, #61637) and Bond RX Automated IHC/ISH Staining System (Leica Biosystems), followed by whole slide imaging by NanoZoomer NDP2.0-HT Digital Slide System (Hamamatsu) and quantitative analysis by HALOTM image analysis software (Indica labs). OX40 IHC scores were also compared to the RNA expression determined by RNAseq. Results. A total of 90 leukemia and lymphoma PDXs were comprehensively evaluated. RNASeq data (human mRNA) indeed revealed OX40 gene expression (cutoff as Log2 (FPKM)>3) in 8.9% of models (8/90 models); on the other hand, IHC data also confirmed these OX40 protein expression; and the IHC scores largely correlated with the RNA data. Together, our data firmly confirmed the expression of OX40 in certain hematological cancers, beyond just in TME. The ongoing studies is investigating the characteristics of the OX40 positive lymphoma/leukemia, and the preliminary data will be presented in the annual conference. Furthermore, we are also using the same assay to explore other cancer types beyond hematological cancers. Conclusion. A tumor associated OX40 expression IHC workflow was preliminarily established and validated to facility the OX40 immunotherapy development. Citation Format: Jie Lin, Xiaolong Tu, Likun Zhang, Hongjiang Yu, Cheng JIang, Tao Yang, Henry Qixiang Li. Immunohistochemistry (IHC) to assess tumor cell-associated OX40 expression in lymphoma PDXs [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 1027.

Abstract 1321: In vivo immune cell programming using mRNA-LNP chimeric antigen receptors

Abstract Introduction: The tumor microenvironment (TME) is a dynamic and multifaceted system that comprises largely myeloid lineage immune cells including monocytes, neutrophils, macrophages, and dendritic cells. Reprogramming the myeloid compartment within the context of cancer therapy presents an attractive strategy to overcome current limitations in solid tumor treatment. We have developed a novel approach that harnesses the natural ability of Myeloid cells to take up mRNA loaded lipid nanoparticles, to reprogram these cells with chimeric antigen receptors (CAR’s) that arm these cells to recognize and kill tumor cells. Methods: Current LNPs technology is sufficient to deliver significant mRNA cargo to numerous cell types, including B cells, T cells and myeloid cells. Leveraging this phenomenon we have developed a new class of chimeric antigen receptors (CARs), using protein biology that imparts targeted immune cell expression and functionality, specifically in Myeloid cells. These CARs harness Immunoreceptor Tyrosine-based Activation Motif (ITAM) signaling adaptors expressed exclusively in immune cells, achieving immune cell-type specific CAR-directed tumor lysis and cytokine responses. With a FcaR I (CD89) based CAR design, we have developed a CAR that is dependent on the common Fc receptor gamma chain, primarily restricted to myeloid cells, for stable expression and function. Results: We have demonstrated specific expression of CD89-based CAR in myeloid cells in mouse and monkey following systemic mRNA/LNP delivery, and in human myeloid cells following in vitro treatment with mRNA/LNPs. Systemic mRNA/LNP delivery of CD89-based CAR has demonstrated robust anti-tumor efficacy against multiple tumor antigens in various xenograft mouse models of human breast cancer, hepatocellular carcinoma, and ovarian cancer. Furthermore, in syngeneic mouse tumor models, robust anti-tumor immune responses, characterized by tumoral infiltration of activated CD8+ T cells, diminished tumor-associated Tregs, systemic enhancement of dendritic cells and anti-tumor IgG production, were observed for treated animals. Conclusions: Taken together, we present CAR designs that achieve expression and function in targeted immune cell populations, capable of eliciting anti-tumor efficacy against multiple target antigens by leveraging innate and adaptive immune responses, and demonstrating a class of novel CAR designs to overcome limitations faced by current in vivo mRNA/LNP delivery strategy. A phase I clinical trial is currently on-going to assess the safety and preliminary efficacy of CD89-based CAR against TROP2+ epithelial malignancies. Citation Format: Shannon Argueta, Franciele K. Melber, Michael Gorgievski, Neha Divangi, Edward Cochran, Thach Chu, Yuxiao Wang, Jian Ding, Daniel Getts. In vivo immune cell programming using mRNA-LNP chimeric antigen receptors [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 1321.

Abstract 4042: Cbl inhibition increases the response of cytotoxic T cells to sub-optimal stimulation and drives enhanced anti-tumor responses alone or in combination with PD-1 blockade

Abstract INTRODUCTION: Herein, we describe a series of experiments to define the unique biology of dual Cbl-b/c-Cbl (Cbl) inhibition in amplifying T cell activation and anti-tumor responses. Through their E3 ubiquitin ligase activity, Cbl-b and c-Cbl have been shown to negatively regulate multiple signaling proteins downstream of the T cell receptor (TCR), suggesting that inhibition may yield enhanced anti-tumor T cell activity. Indeed, using a dual Cbl-b/c-Cbl small molecule inhibitor, we demonstrate that Cbl blockade enhances T cell activity under standard and sub-optimal stimulation conditions. Moreover, Cbl inhibition increases mouse syngeneic tumor control alone or in combination with α-PD-1, as described below. METHODS: Human CD8+ T cells were isolated from healthy human blood and activated using α-CD3 or α-CD3/CD28 stimulation. Mouse OT-I splenocytes were stimulated with SIINFEKL or lower affinity ovalbumin peptides and cytokine production was assessed. For tumor studies, mice were inoculated with syngeneic cancer cells and were treated with Cbl inhibitor (30 mg/kg, QD) +/- α-PD-1 (10 mg/kg, Q5D) starting at a tumor volume of 75 mm3. RESULTS: Human CD8+ T cells, stimulated in the presence of a Cbl inhibitor, demonstrated a concentration-dependent increase in activation hallmarks including cytokine secretion (IL-2, IFN-γ and granzyme B), proliferation, and the presence of highly polyfunctional cells. Furthermore, using serially stimulated CD8+ T cells with reduced overall activity, Cbl inhibition showed a marked enhancement of IFN-γ production and cytotoxicity compared to controls upon re-stimulation. We then used an antigen-specific T cell assay where OT-I splenocytes were stimulated by peptides with optimal or reduced TCR affinities, Cbl inhibition produced significantly higher IFN-γ levels with all peptides relative to controls demonstrating that Cbl inhibition can increase the response of T cells to sub-optimal stimulation. Using mouse CT26 and MC38 tumor models, Cbl inhibition was able to significantly enhance tumor control in combination with α-PD-1 and as a monotherapy in CT26 tumors. Notably, a greater number of tumor antigen-specific cytotoxic T cells were found in Cbl inhibitor-treated tumors. Higher expression levels of the co-stimulatory molecule CD226 and the TCR in these anti-tumor cells were also observed, demonstrating the unique biology of Cbl inhibition in enhancing anti-tumor responses. Finally, Cbl inhibition had no effect on the growth or survival of cancer cells in vitro consistent with the notion that enhanced tumor control was driven by immune activity. CONCLUSIONS: Altogether, these data demonstrate that pharmacologic Cbl inhibition robustly enhances T cell activity in vitro and in vivo and provide a mechanistic rationale for targeting Cbl-b/c-Cbl to amplify anti-tumor immune responses. Citation Format: Livia Yamashiro, Sachie Marubayashi, Rameshwari Rayaji, Anudari Battsooj, Karim Mrouj, Hema Singh, Yue Tong Lee, Ada Chen, Lunda Shen, Artur Mailyan, Kai Yu, Masha Podunavac, Julie Yu, Dongdong Liu, Clayton Hardman, Johnny Yin, Srikanth Gangam, Ken Lawson, Manmohan Leleti, Matthew Walters, Daniel DiRenzo. Cbl inhibition increases the response of cytotoxic T cells to sub-optimal stimulation and drives enhanced anti-tumor responses alone or in combination with PD-1 blockade [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 4042.

Enhancement of sustainable bioethanol production from microorganisms isolated from molasses and venasses

Enhancement of sustainable bioethanol production from microorganisms isolated from molasses and venasses

25-hydroxycholesterol triggers antioxidant signaling in mouse atria

Oxysterol, 25-hydroxycholesterol (25HC), is a potent regulator of immune reactions, its synthesis greatly increases by macrophages during inflammation. We hypothesize that 25HC can have cardioprotective effects by limiting consequences of excessive β-adrenoceptor (βAR) stimulation, particularly reactive oxygen species (ROS) production, in mouse atria. Isoproterenol, a βAR agonist, increased extra- and intracellular levels of ROS. This enhancement of ROS production was suppressed by NADPH oxidase antagonists as well as 25HC. Inhibition of β3ARs, Gi protein and protein kinase Cε prevented the effect of 25HC on isoproterenol-dependent ROS synthesis. Furthermore, 25HC suppressed isoproterenol-induced lipid peroxidation and mitochondrial ROS generation as well as ROS-dependent component of positive inotropic response to isoproterenol. Additionally, 25HC decreased mitochondrial ROS production and lipid peroxidation induced by antimycin A, a mitochondrial poison. Thus, 25HC exerts antioxidant properties alleviating mitochondrial dysfunction-induced and βAR-dependent cardiac oxidative damage. In the latter case, 25HC can act via signaling mechanism engaging β3ARs, Gi protein and protein kinase Cε.

Nurturing family business resilience through strategic supply chain management

PurposeThe study aspires to enhance comprehension of the intricate interplay between supply chain management (SCM) and resilience in family businesses, thereby offering valuable insights to managers and policymakers endeavouring to foster resilience in uncertain environments.Design/methodology/approachCommencing from the premise that family businesses (FBs) prioritize the preservation of socio-emotional wealth (SEW) when formulating strategic decisions, this study endeavours to advance understanding of supply chain practices adopted by FBs and their direct impact on resilience during crisis situations or economically challenging periods. Through an exploratory case study of nine FBs, the present research reveals four pivotal strategies in SCM that contribute to their resilience: (i) reorganization of inventory management; (ii) cultivating close relationships with suppliers; (iii) emphasizing product quality and customer retention; and (iv) implementing cost reduction measures to bolster resilience. The aim of the study is to provide an in-depth understanding of the intricate interplay between SCM and resilience in FBs, thereby offering valuable insights to managers and policymakers endeavouring to foster resilience in uncertain environments.FindingsOur approach offers a theoretical framework for SCM aligned with prior research on the interplay between characteristics of family businesses and resilience strategies. Furthermore, this paper illustrates how factors such as the emphasis on high-quality products and services by family businesses contribute to achieving non-economic objectives that owners adopt to reconcile family and business needs, creating intrinsic added value for the company. It reveals various challenges in SCM, including inventory organization changes, supplier closures and the significance of customer retention. Family businesses are implementing product and technology enhancements and leveraging digitization to enhance supply chain processes.Originality/valueThis paper contributes significantly to the field of FBs by highlighting the crucial role of SCM in enhancing business resilience during crises. It empirically examines how the SEW characteristics of FBs influence the reconfiguration of their supply chains to enhance resilience, presenting a theoretical model for this context. Our theoretical framework employs an SEW perspective to elucidate how FBs respond to the challenges posed by the COVID-19 pandemic by adapting their SCM processes to safeguard their social and emotional legitimacy, organizational visibility and reputation. These adaptations gain particular relevance during crises or turbulent conditions, potentially leading to alterations in how FBs formulate their supply chain strategies and manage supply chain-related processes.

Financial impact of incorporating deep learning reconstruction into magnetic resonance imaging routine

PurposeArtificial intelligence and deep learning solutions are increasingly utilized in healthcare and radiology. The number of studies addressing their enhancement of productivity and monetary impact is, however, still limited. Our hospital has faced a need to enhance MRI scanner throughput, and we investigate the utility of new commercial deep learning reconstruction (DLR) algorithm for this purpose. In this work, a multidisciplinary team evaluated the impact of the widespread deployment of a new commercial deep learning reconstruction (DLR) algorithm for our magnetic resonance imaging scanner fleet. MethodsOur analysis centers on the DLR algorithm's effects on patient throughput and investment costs, contrasting these with alternative strategies for capacity expansion—namely, acquiring additional MRI scanners and increasing device utilization on weekends. We provide a framework for assessing the financial implications of new technologies in a trial phase, aiding in informed decision-making for healthcare investments. ResultsWe demonstrate substantial reductions in total operating costs compared to other capacity-enhancing methods. Specifically, the cost of adopting the deep learning technology for our entire scanner fleet is only 11 % compared to procuring an additional scanner and 20 % compared to the weekend utilization costs of existing devices. ConclusionsProcuring DLR for our existing five-scanner fleet allows us to sustain our current MRI service levels without the need for an additional scanner, thereby achieving considerable cost savings. These reductions highlight the efficiency and economic viability of DLR in optimizing MRI service delivery.

Conditional expression of FumA in Aspergillus niger enhances synthesis of L-malic acid.

Currently, the L-malic acid titer achieved through Aspergillus niger fermentation reaches 201 g/L, meeting industrial demands satisfactorily. However, the co-presence of structurally similar fumaric acid and succinic acid in fermentation products suggests a theoretical potential for further improvement in L-malic acid production. In the tricarboxylic acid cycle, fumarate reductase mediates the conversion of succinic acid to fumaric acid. Subsequently, fumarase catalyzes the conversion of fumaric acid to L-malic acid. Notably, both enzymatic reactions are reversible. Our investigation revealed that A. niger contains only one mitochondria-located fumarase FumA. Employing CRISPR-Cas9 technology, we performed a replacement of the fumA promoter with a doxycycline-induced promoter Tet. Under non-inducing condition, the conditional strain exhibited increased levels of fumaric acid and succinic acid. It strongly suggests that FumA mainly promotes the flow of fumaric acid to L-malic acid. Furthermore, a promoter PmfsA that is exclusively activated in a fermentation medium by calcium carbonate was identified through RNA-sequencing screening. Utilizing PmfsA to regulate fumA expression led to a 9.0% increase in L-malic acid titer, an 8.75% increase in yield (glucose to L-malic acid), and an 8.86% enhancement in productivity. This research serves as a significant step toward expediting the industrialization of L-malic acid synthesis via biological fermentation. Additionally, it offers valuable insights for the biosynthesis of other organic acids.IMPORTANCEThis study focuses on enhancing L-malic acid synthesis by modifying the tricarboxylic acid cycle within the mitochondria of Aspergillus niger. We emphasize the significant role of fumarase in converting fumaric acid into L-malic acid, enhancing our understanding of metabolic pathways in A. niger. The precise regulation of fumA is highlighted as a key factor in enhancing L-malic acid production. Furthermore, this research introduces a stringent conditional promoter (PmfsA), exclusively activated by CaCO3. The utilization of PmfsA for fumA expression resulted in heightened L-malic acid titers. The progress in metabolic engineering and bioprocess optimization holds promise for expediting industrial L-malic acid synthesis via biological fermentation. Moreover, it carries implications for the biosynthesis of various other organic acids.

Alkali Polymer Flooding of a Romanian Field Containing Viscous Reactive Oil.

The study demonstrates the significant enhancement in oil production from a Romanian oil field using alkali-polymer (AP) flooding for reactive viscous oil. We conducted comprehensive interfacial tension (IFT) measurements across various alkali and AP concentrations, along with phase behavior assessments. Micromodel flooding experiments were used to examine pore-scale effects and select appropriate chemical concentrations. We tested displacement efficiency at the core level and experimented with different sequences and concentrations of alkali and polymers to minimize costs while maximizing the additional recovery of reactive viscous oil. The IFT analysis revealed that saponification at the oil-alkali interface significantly lowers IFT, but IFT gradually increases as soap diffuses away from the interface. Micromodels indicated that polymer or alkali injection alone achieve only minimal incremental recovery beyond waterflooding. However, AP flooding significantly enhanced incremental oil recovery by efficiently moving the mobilized oil with the viscous fluid and increasing exposure of more oil to the alkali solution. Coreflood experiments corroborated these findings. We also explored how divalent cations influence polymer concentration selection, finding that softening the injection brine significantly increased the viscosity of the AP slug.

Strategic Oversight and Quality Validation in Spice Production Enhancement

This study examines the effectiveness of a comprehensive control management and quality control system in the seasoning production process, focusing specifically on a menu-specific seasoning variant (MSS). The methodology encompassed various stages including material acceptance, sieving, mixing, and final packaging, integrated with stringent control measures such as equipment and facility cleaning, material separation, and machine maintenance. Quality control tests conducted included sodium chloride content, pH analysis, seal strength, and bubble tests. Results indicate that these systematic processes significantly enhance product quality by ensuring compliance with predefined quality standards. The implications of this study suggest that rigorous process oversight and detailed quality assessments can markedly improve the reliability and safety of food seasoning products, providing a framework for industry-wide best practices. Highlights: Process Management: Systematic control measures, including equipment and facility cleaning, ensure a contamination-free production environment. Quality Control: Rigorous testing, including sodium chloride content and pH analysis, guarantees that the products meet established quality and safety standards. Product Compliance: Implementation of advanced control and quality assurance protocols ensures that products consistently comply with industry standards. Keywords: Instant Seasoning, Quality Control, Safety Standards

Economic and Performance Analysis of Modified Solar Distillation System Coupling Different Integrations Using Carbon Quantum Dot Nanoparticles: Generalized Thermal Model

Abstract Clean drinking water and electricity production utilizing non-conventional sources of energy is the global demand for sustainable development. Ultrafast heat transfer fluids have delivered impressive results in photovoltaic (PV)-integrated solar thermal systems, in recent times. Efforts have been made for the productivity and electricity augmentation of solar still equipped with helically coilled heat exchanger and coupled with different integrations, viz., (a) partially covered N-photovoltaic thermal compound parabolic concentrator (N-PVT-CPC), (b) partially covered N-photovoltaic thermal flat plate collector (N-PVT-FPC), (c) N-FPC-CPC, and (d) N-flat plate collector (N-FPC). System design has also been modified by adding a roof-top semi-transparent PV module and built-in passive copper condenser (circulation mode), and effect of carbon quantum dots (CQDs) water-based nanofluids, nanoparticles volume concentration, and packing factor (βc) of the PV module has been studied by developing generalized thermal modeling of the system (special cases). Overall, 41.1%, 21.52%, 22.01%, and 10.01% rise in evaporative HTCs is observed in FPC-CPC, PVT-CPC, FPC, and PVT-FPC integrations, respectively. Thermal exergy is found to be higher for FPC-CPC integration, and it follows the enhancement order as FPC-CPC (max-0.147 kW) > PVT-CPC (0.088 kW) > FPC (0.038 kW) > PVT-FPC (0.028 kW). In reference to the base fluid, significant enhancement in the daily productivity is observed for FPC-CPC (10.9%) and PVT-CPC (5.16%) integrations using CQD-NPs. The production cost of potable water has also been estimated for all the cases for n = 30 and n = 50 years life span and i = 4% and 8% interest rates, and it is found to be the lowest (0.014 $/L) for FPC-CPC integration using CQD-NPs (n = 30 years, i = 4%).

Multiple-pathways light modulation in Pleurosigma strigosum bi-raphid diatom.

Ordered, quasi-ordered, and even disordered nanostructures can be identified as constituent components of several protists, plants and animals, making possible an efficient manipulation of light for intra- and inter- species communication, camouflage, or for the enhancement of primary production. Diatoms are ubiquitous unicellular microalgae inhabiting all the aquatic environments on Earth. They developed, through tens of millions of years of evolution, ultrastructured silica cell walls, the frustules, able to handle optical radiation through multiple diffractive, refractive, and wave-guiding processes, possibly at the basis of their high photosynthetic efficiency. In this study, we employed a range of imaging, spectroscopic and numerical techniques (including transmission imaging, digital holography, photoluminescence spectroscopy, and numerical simulations based on wide-angle beam propagation method) to identify and describe different mechanisms by which Pleurosigma strigosum frustules can modulate optical radiation of different spectral content. Finally, we correlated the optical response of the frustule to the interaction with light in living, individual cells within their aquatic environment following various irradiation treatments. The obtained results demonstrate the favorable transmission of photosynthetic active radiation inside the cell compared to potentially detrimental ultraviolet radiation.

Lactiplantibacillus plantarum and Komagataeibacter europaeus enhance energy metabolism, acetic acid and aromatic amino acids catabolism flux in cider vinegar fermentation

Fruit vinegar is typically produced through a two-stage submerged fermentation involving Saccharomyces cerevisiae (alcohol fermentation) and Acetobacter pasteurianus (acetic acid fermentation). In order to enhance its flavor and nutritional properties, Lactiplantibacillus plantarum and Komagataeibacter europaeus were introduced into the respective stages. The fermentation process was monitored to assess their individual and combined effects, while non-targeted metabolomic analysis aided amino acids and organic acids analysis was applied to characterize the metabolic profiles and identify the differential metabolites.Lactic acid accumulated by L. plantarum, enhanced energy metabolism and ethanol respiratory chain, thereby promoting the K. europaeus growth. In the presence of K. europaeus, A. pasteurianus biomass increased 36–57%, leading to elevated acetic acid synthesis. The co-fermentation systems, especially those containing K. europaeus, exhibited enhanced conversion of shikimic acid to aromatic amino acids and their derivatives such as ferulic acid, isoferulic acid, serotonin, l-kynuanine, phenylacetaldehyde and benzaldehyde. The co-fermentation with K. europaeus and L. plantarum resulted in an enhancement of ethyl phenylacetate, ethyl lactate and ethyl hexanoate production, thereby intensifying the fruity aroma profile. The findings presented here contributed to a deeper comprehension of bioconversion mechanism involving multiple species in cider vinegar fermentation processes.

Multi-mode enhancement and Co-fermentation mechanism of 2-phenylethanol production by Kluyveromyces marxianus

As a valuable aromatic alcohol, 2-phenylethanol has been widely used in the fields of food, medicine and chemistry, but its production has become a key factor limiting microbial industrialisation. Therefore, atmospheric and room temperature plasma (ARTP) mutagenesis was applied to improve host cell tolerance of 2-phenylethanol, and 2-phenylethanol production was increased from 2.23 g/L to 2.51 g/L in Kluyveromyces marxianus. Fermentation conditions of K. marxianus and Pediococcus lactis co-culture were optimised and production of 2-phenylethanol was further increased by 42.63%, reaching 3.58 g/L. Metabonomics analysis revealed that the cell wall of P. lactis could effectively activate Ehrlich pathway-related metabolism and upregulate arginine synthesis pathways to enhance the cellular robustness of K. marxianus, thereby improving the efficiency of 2-phenylethanol synthesis, and providing a novel strategy for cost-effective 2-phenylethanol production.

Enhancement of methane production from anaerobic digestion of Erigeron canadensis via O2-nanobubble water supplementation

Malignant invasive Erigeron canadensis, as a typical lignocellulosic biomass, is a formidable challenge for sustainable and efficient resource utilization, however nanobubble water (NBW) coupled with anaerobic digestion furnishes a prospective strategy with superior environmental and economic effectiveness. In this study, influence mechanism of various O2-NBW addition times on methanogenic performance of E. canadensis during anaerobic digestion were performed to achieve the optimal pollution-free energy conversion. Results showed that supplementation of O2-NBW in digestion system could significantly enhance the methane production by 10.70–16.17%, while the maximum cumulative methane production reached 343.18 mL g−1 VS in the case of one-time O2-NBW addition on day 0. Furthermore, addition of O2-NBW was conducive to an increase of 2–90% in the activities of dehydrogenase, α-glucosidase and coenzyme F420. Simultaneously, both facultative bacteria and methanogenic archaea were enriched as well, further indicating that O2-NBW might be responsible for facilitating hydrolytic acidification and methanogenesis. Based on Kyoto Encyclopedia of Genes and Genomes (KEGG) cluster analysis, provision of O2-NBW enhanced the metabolism of carbohydrate and amino acid, translation as well as membrane transport of bacteria and archaea. This study might offer the theoretical guidance and novel insights for efficient recovery of energy from lignocellulosic biomass on account of O2-NBW adhibition in anaerobic digestion system, progressing tenor of carbon-neutral vision.

Multi-objective simulated annealing algorithm for robotic mixed-model two-sided assembly line balancing with setup times and multiple constraints

Robotic mixed-model two-sided assembly lines (RMTALs) have become increasingly prevalent in manufacturing industries for productivity enhancement. However, only limited attention has been paid to the RMTAL balancing problems that contain both setup times and multiple constraints such as positional constraints, zoning constraints, and synchronism constraints. Given the coexistence of these aspects in real-world scenarios, this study aims to address the RMTAL balancing problem with setup times and the mentioned constraints. To formulate the problem, a mixed integer programming model is proposed to optimize cycle time and total energy consumption. To handle multiple constraints and setup times, a new method based on four-vector encoding is introduced. This method addresses the three constraints during the encoding phase using four intervals and five rules, while setup times are processed during the decoding phase. To solve the problem, a Pareto entropy-based two-mode multi-objective simulated annealing algorithm is developed. The algorithm employs variable neighborhood descent algorithm with switchable objectives to generate the initial Pareto archive. Subsequently, it selects the initial solution with minimal potential in the Pareto archive, alternating between two search modes of exploration and exploitation based on the entropy difference of Pareto solutions. Comparative experiments with four state-of-the-art algorithms on benchmark instances of varying scales demonstrate that the proposed algorithm outperforms the others in over 93% of instances for the hypervolume ratio and inverted generational distance metrics.

Novel 4th-generation phytase improves broiler growth performance and reduces woody breast severity through modulation of muscle glucose uptake and metabolism.

The objective of the present study was to determine the effect of a novel (4th generation) phytase supplementation as well as its mode of action on growth, meat quality, and incidence of muscle myopathies. One-day old male broilers (n = 720) were weighed and randomly allocated to 30 floor pens (24 birds/pen) with 10 replicate pens per treatment. Three diets were fed from hatch to 56- days-old: a 3-phase corn-soy based diet as a positive control (PC); a negative control (NC) formulated to be isocaloric and isonitrogenous to the PC and with a reduction in Ca and available P, respectively; and the NC supplemented with 2,000 phytase units per kg of diet (NC + P). At the conclusion of the experiment, birds fed with NC + P diet were significantly heavier and had 2.1- and 4.2-points better feed conversion ratio (FCR) compared to birds offered NC and PC diets, respectively. Processing data showed that phytase supplementation increased live weight, hot carcass without giblets, wings, tender, and skin-on drum and thigh compared to both NC and PC diets. Macroscopic scoring showed that birds fed the NC + P diet had lower woody breast (WB) severity compared to those fed the PC and NC diets, however there was no effect on white striping (WS) incidence and meat quality parameters (pH, drip loss, meat color). To delineate its mode of action, iSTAT showed that blood glucose concentrations were significantly lower in birds fed NC + P diet compared to those offered PC and NC diets, suggesting a better glucose uptake. In support, molecular analyses demonstrated that the breast muscle expression (mRNA and protein) of glucose transporter 1 (GLUT1) and glucokinase (GK) was significantly upregulated in birds fed NC + P diet compared to those fed the NC and PC diets. The expression of mitochondrial ATP synthase F0 subunit 8 (MT-ATP8) was significantly upregulated in NC + P compared to other groups, indicating intracellular ATP abundance for anabolic pathways. This was confirmed by the reduced level of phosphorylated-AMP-activated protein kinase (AMPKα1/2) at Thr172 site, upregulation of glycogen synthase (GYS1) gene and activation of mechanistic target of rapamycin and ribosomal protein S6 kinase (mTOR-P70S6K) pathway. In conclusion, this is the first report showing that in-feed supplementation of the novel phytase improves growth performance and reduces WB severity in broilers potentially through enhancement of glucose uptake, glycolysis, and intracellular ATP production, which used for muscle glycogenesis and protein synthesis.

Outdoor biohydrogen production by thermotolerant Rhodopseudomonas pentothenatexigens KKU-SN1/1 in a cluster of ten bioreactors system.

In tropical regions, the viability of outdoor photo-fermentative biohydrogen production faces challenges arising from elevated temperatures and varying light intensity. This research aimed to explore how high temperatures and outdoor environments impact both biohydrogen production and the growth of purple non-sulfur bacteria. Our findings revealed the potential of Rhodopseudomonas spp. as a robust outdoor hydrogen-producing bacteria, demonstrating its capacity to thrive and generate biohydrogen even at 40°C and under fluctuating outdoor conditions. Rhodopseudomonas harwoodiae NM3/1-2 produced the highest cumulative biohydrogen of 223mL/L under anaerobic light conditions at 40°C, while Rhodopseudomonas harwoodiae 2M had the highest dry cell weight of 2.93g/L. However, R. harwoodiae NM3/1-2 demonstrated the highest dry cell weight of 3.99g/L and Rhodopseudomonas pentothenatexigens KKU-SN1/1 exhibited the highest cumulative biohydrogen production of 400mL/L when grown outdoors. In addition, the outdoor enhancement of biohydrogen production was achieved through the utilization of a cluster of ten bioreactors system. The outcomes demonstrated a notable improvement in biohydrogen production efficiency, marked by the highest daily biohydrogen production of 493mL/Ld by R. pentothenatexigens KKU-SN1/1. Significantly, the highest biohydrogen production rate was noted to be 17 times greater than that observed in conventional batch production methods. This study is the first to utilize R. pentothenatexigens and R. harwoodiae for sustained biohydrogen production at high temperatures and in outdoor conditions over an extended operational period. The successful utilization of a clustered system of ten bioreactors demonstrates potential to scale-up for industrial biohydrogen production.

Multidirectional pathway engineering of Mycobacterium neoaurum LY-2 for the enhancement of androstenedione production from phytosterols

Androstenedione (ADD), an important precursor for steroid hormone drug synthesis, can be obtained through the transformation of phytosterols by members of the Mycobacteria genus. In this study, multidirectional pathway engineering of Mycobacterium neoaurum LY-2 was applied to improve ADD production from phytosterols. Initially, the nonessential gene KasB encoding a cell wall β-ketoacyl acyl carrier protein synthase in Mycobacterium neoaurum LY-2 was knocked out, and the production of ADD was increased by 21.0%. Then, the key enzymes involved in ADD synthetic metabolism were systematically explored. These assays revealed that propionyl coenzyme A carboxylase (PCC) and Hsd4A played important roles in the synthesis of ADD. In addition, the heterologous expression of Vitreoscilla hemoglobin (VHb), a protein with oxygen-binding capacity, was successfully realized in Mycobacterium neoaurum LY-2, and the ADD concentration reached 4.42 g/L. Finally, the recombinant strain LY-2-ΔKasB-hsd4A-pcc-vgb with high ADD production was constructed by combining the above strategies. Through optimization of conversion conditions, the recombinant strain LY-2-ΔKasB-hsd4A-pcc-vgb produced an ADD concentration of 11.12 g/L when adding 20 g/L phytosterols.

A Laboratory Experimental Study on Enhancing the Oil Recovery Mechanisms of Polymeric Surfactants.

In order to evaluate the physical and chemical properties of polymer surfactants and analyze their oil displacement mechanisms, three types of poly-surfactant used in the Daqing oil field were chosen to be researched, and the oil displacement effects were studied using poly-surfactants of different viscosity, dehydrating rate, and core permeability. The main purpose is to determine the reasonable range of different characteristic indexes of polymeric surfactant flooding. The oil displacement effect of 15 cores was analyzed, and the effects of viscosity, the dehydrating rate of emulsion, and permeability on EOR (Enhanced Oil Recovery) were analyzed. The oil displacement mechanisms of polymeric surfactants were researched using a photolithographic glass core. This paper explores the mechanism underlying production enhancement as an EOR target, while simultaneously conducting laboratory tests to assess the physical and chemical properties of polymeric surfactants. The poly-surfactant agents exhibit a notable increase in viscosity, with the optimal displacement effect observed at a core effective permeability exceeding 400 mD, resulting in a potential EOR of 15% or higher. Moreover, at a viscosity ranging between 40 and 70 mPa·s, the total EOR can reach 73%, with the peak efficiency occurring at a viscosity of 60 mPa·s. The water loss rate of the emulsion, ranging between 30% and 70%, achieves optimal performance at 50%. The poly-surfactants' higher viscosity extends the oil sweep area, enhancing recovery efficiency, and noticeably reducing residual oil compared to water flooding. During poly-surfactant flooding, a substantial amount of residual oil is extracted and transformed into droplets. The rapid emulsification of the polymeric surfactant solution with crude oil forms a stable emulsion, contributing to its significant oil recovery effect. This research provides valuable technical support for EOR in thin and low-quality reservoirs of onshore multi-layered sandstone reservoirs.