Valorization of sewage sludge incineration ash as a novel soilless growing medium for urban agriculture and greenery.

Limited open areas for urban agriculture and greenery have led to the search for innovative, sustainable growing media to strengthen the food supply and improve atmospheric quality for a resilient city. Rampant land developments have caused soil to become increasingly scarce. Sewage sludge incineration ash (SSIA), the by-product of waste-to-energy (WtE) incineration of sewage sludge, is a major municipal waste containing phosphorus-fertilizing nutrients. For the first time, we investigated the novel application of SSIA as a soilless plant-growing medium with built-in fertilizer. SSIA outperformed topsoil in bulk density, water-holding capacity, porosity, and nutrient content. However, it was found that SSIA has a high salinity and should be treated first. Wheatgrass (Triticum aestivum L.), a fast-growing glycophyte, thrived in the desalinated SSIA, showing growth and nutrient content comparable to the topsoil case. Simultaneously, it demonstrated phytoremediation. The SSIA residue was then recycled into cementitious materials, using desalinating water for mixing. SSIA upcycle into a growing medium facilitates urban resource management by utilizing nutrients in sewage waste for eco-friendly plant cultivation, benefiting urban agriculture and greenery. It is also a prudent valorization step before further recycling SSIA to reduce landfill requirements.

Excess copper promotes an increase in the concentration of metabolites in Tridax procumbens L.

The study investigated the effects of cultivating Tridax procumbens in hydroponic conditions with different concentrations of copper ions, aiming to understand the physiological changes and the impact on the biosynthesis of secondary metabolites. The treatments consisted of a completely randomized design, with five increasing concentrations of copper (T0 = 0.235, T1 = 12.5, T2 = 25, T3 = 50, T4 = 100 µmol L-1 of Cu), under controlled conditions for 36 days. Analysis of bioactive compounds in leaves was performed by HPLC-DAD and ESI-MS. Several phenolic compounds, alkaloids, phytosterols and triterpenoids were identified, demonstrating the plant's metabolic plasticity. The highest dose of copper (100 µmol L-1) significantly promoted voacangine, the most predominant compound in the analyses. Notably, 66.7% of the metabolites that showed an increase in concentration, were phenolic compounds. Furthermore, treatments with 12.5 and 25 µmol L-1 of copper were identified as promoting the biosynthesis of phytosterols and triterpenoids. These biochemical adaptations can play a fundamental role in the survival and development of plants in environments contaminated by metals, and from this it is possible to determine cultivation techniques that maximize the biosynthesis of the compound of interest.

Mutagenic effect of ultraviolet radiation and trimethyl psoralen in Mycoplasma towards minimal genome.

Mycoplasma, solely culturable bacteria with the smallest genome, is an important organism to understand the minimal form of life. Mutagenesis using mutagens is a useful methodology for understanding the essential regions of genomic information. Ultraviolet light and trimethyl psoralen are mutagens known to induce various mutations; the latter is reported to specifically induce deletions in nematodes. However, their mutagenic effects on mycoplasma are not known. Here, we exposed Metamycoplasma salivarium to ultraviolet (UV) light or trimethyl psoralen and UV as mutagens, and analyzed the mutational pattern after several rounds of serial cultivation ranging from 34 to 56 for different lineages. Our results showed that more deletions, but fewer point mutations, were induced with TMP and UV-A than with UV alone, indicating the usefulness of TMP in inducing deletions. In addition, we compared our results with mutational data from other studies, which suggested that both TMP-UVA and UV exposure induced point mutations that were highly biased toward C to T and G to A transitions. These data provide useful basic knowledge for mutational studies on M. salivarium.

Microalgal production and nutrient recovery under mixotrophic mode using cheese whey permeate.

Mixotrophic microalgal solutions are efficient nutrient recovery methods, with potential to prolong the cultivation seasons in temperate climates. To improve operation sustainability, the study used landfill leachate for nitrogen and whey permeate for phosphorus and organic carbon. A non-axenic polyculture, dominated by green algae, was cultivated in mixotrophic mode on glucose or whey permeate compared to a photoautotrophic control in outdoor large-scale raceway ponds during Nordic spring and autumn. The whey permeate treatment had the highest algal growth rate and productivity (0.48 d-1, 183.8 mg L-1 d-1), nutrient removal (total nitrogen: 21.71 mg L-1 d-1, total phosphorus: 3.05 mg L-1 d-1) and recovery rate (carbon: 85.19 mg L-1 d-1, nitrogen: 17.01 mg L-1 d-1, phosphorus: 2.58 mg L-1 d-1). When grown in whey permeate, algal cultures demonstrated consistent productivity and biochemical composition in high (spring) and low light conditions (autumn), suggesting the feasibility of year-round production in Nordic conditions.

Synergistic effects of bivalve and microalgae co-cultivation on carbon dynamics and water quality.

Aquaculture of bivalve shellfish and algae offers significant ecological benefits, yet the complex interactions between these organisms can substantially impact local carbon dynamics. This study investigated the effects of co-culturing four intertidal bivalve species Pacific oysters (Crassostrea gigas), Manila clams (Ruditapes philippinarum), Chinese clams (Cyclina sinensis), and hard clams (Mercenaria mercenaria) with microalgae (Isochrysis galbana) on specific water quality parameters, including total particulate matter (TPM), total organic matter (TOM), dissolved inorganic carbon (DIC), dissolved carbon dioxide (dCO2), dissolved oxygen (DO), and ammonium (NH4+) concentrations. The bivalves were divided into smaller and larger groups and cultured under two conditions: with algae (WP) and without (NP), along with matched controls. Total particulate matter (TPM), total organic matter (TOM), dissolved oxygen (DO), ammonium nitrogen (NH4+), dissolved inorganic carbon (DIC), and CO2 (dCO2) were measured before and after 3-h cultivation. Results revealed species-specific impacts on water chemistry. C. gigas, C. sinensis and R. philippinarum showed the strongest reduction in DIC and dCO2 in WP groups, indicating synergistic bioremediation with algae. M. mercenaria notably reduced TPM, highlighting its particle carbon sequestration potential. DO concentrations decreased in most WP or NP groups, reflecting respiration of the cultured bivalves or microalgae. NH4+ levels also declined for most species, indicating nitrogen assimilation by these creatures. Overall, the bivalve size significantly impacted carbon and nitrogen processing capacities. These findings reveal species-specific capabilities in regulating water carbon dynamics. Further research should explore integrating these bivalves in carbon-negative aquaculture systems to mitigate environmental impacts. This study provides valuable insights underlying local carbon dynamics in shallow marine ecosystems.

In-situ rapid cultivation of aerobic granular sludge in A/O bioreactor by using Ca(ClO)2 pretreating sludge.

The efficient utilization of residual sludge and the rapid cultivation of aerobic granular sludge in continuous-flow engineering applications present significant challenges. In this study, aerobic granular cultivation was fostered in a continuous-flow system using Ca(ClO)2-sludge carbon (Ca-SC). Ca-SC retained the original sludge properties, contributing to granular growth in an A/O bioreactor. By day 40, the granule diameters increased to 0.8 mm with the SVI30 decreased by 2.7 times. Moreover, Ca-SC facilitated protein secretion, reaching 98.06 mg/g VSS and enhanced the hydrophobicity to 68.4 %. The continuous-flow aerobic granular sludge exhibited a nutrient removal rate above 90 %. Furthermore, Tessaracoccus and Nitrospira were enriched to promote granular formation and nitrogen removal. The residual sludge was carbonized and reused in the traditional wastewater treatment process to culture granular sludge in situ, aiming to achieve "self-production and self-consumption" of sludge and promote the innovative model of "treating waste with waste" in urban sewage environmental restoration.

Long-term changes of methane emissions from rice cultivation during 2000 - 2060 in China: Trends, driving factors, predictions and policy implications.

Regional budget assessments of methane (CH4) are critical for future climate and environmental management. CH4 emissions from rice cultivation (CH4-rice) constitute one of the most significant sources. However, previous studies mainly focus on historical emission estimates and lack consideration of future changes in CH4-rice under climate change or anthropogenic policy intervention, which hampers our understanding of long-term trends and the implementation of targeted emission reduction efforts. This study investigates the spatiotemporal variations of CH4-rice over the past two decades, using an integrated method to identify the major drivers and predict future emissions under climate change scenarios and policy perspectives. Results indicate that the CH4-rice emissions in China ranged between 6.21 and 6.57 Tg yr-1 over the past two decades, with a spatial distribution characterized by decreases in the south and increases in the north, associated with economic development, dietary shifts, technological advancements, and climate change. Factors such as the rate of straw added (RSA), fertilization, soil texture, temperature, and precipitation significantly influence CH4 emissions per unit rice production (CH4-urp), with RSA identified as the most significant tillage management factor, explaining 32 % of the variance. Lowering RSA to 8 % is beneficial for reducing CH4-urp. Scenario analysis indicates that under policies focusing on production or demand, CH4-rice is expected to increase by 0.3 % to 5.6 %, while adjusting RSA can reduce CH4-rice by 9.4 % to 10.0 %. Structural adjustments and regional cooperation serve as beneficial starting points for controlling and reducing CH4-rice in China, while optimizing industrial layouts contributes to regional development and CH4-rice control. Implementing policies related to maintaining field and crop yields can achieve a balance between rice supply and demand ahead of schedule. Dynamic adjustment of rice cultivation based on supply-demand balance can effectively reduce CH4-rice from excess rice production. By 2060, the reduction effect could reach 8.95 %-12.01 %. Introducing policy-driven tillage management measures as reference indicators facilitates the reduction of CH4-rice.

In vitro cultivation of adherent cells on microscope slides for downstream applications.

In vitro studies with cultured cells are often conducted as an important part of basic research. Adherent cells are typically cultivated in flasks or trays, for which cell staining and subsequent visualization become impractical. We here present a simple step-by-step method for growing adherent cells directly on glass microscope slides, using low-cost equipment readily available in most laboratories. Most parameters such as type of microscope slide (e.g. surface coating), cell seeding concentrations and incubation times can be adjusted according to cell line characteristics and experimental aims, reflecting the methods' flexibility. Through our experiments, microscope slides proved to provide an acceptable surface for cell adhesion and growth of the tested cell lines, as well as being robust and functional with respect to downstream procedures. The method can potentially be combined with different techniques for visualization of experimental effects, such as histological staining methods, fluorescent staining, and immunochemistry. In our method development we have successfully cultivated three different cell lines directly on microscope slides - Atlantic salmon kidney cells (ASK), rainbow trout gill cells (RTgill-W1), and human cancerous lung cells (A549) - and subjected them to various experimental treatments. Finally, as proof-of-concept we provide examples of successful histological staining of the fixed cells. Experimental design in short:•Cultivate cells and calculate cell concentration•Seed a small volume of growth medium with an appropriate number of cells on microscope slide in an area confined by hydrophobic marker•Let cells adhere over night before adding more growth medium or directly conducting experiments and fixing cells for downstream applications.

Prognostic value of 24-hour cultivation of peritoneal fluid to distinguish complicated from uncomplicated acute appendicitis: a prospective cohort study.

BACKGROUND: The distinction between complicated and uncomplicated acute appendicitis (AA) is important as it guides postoperative antibiotic treatment. A diagnosis based on intraoperative findings is imprecise and standard cultivation of peritoneal fluid is generally time-consuming with little clinical benefit. The aim of this study was to examine if cultivation of peritoneal fluid in acute appendicitis could reliably detect bacteria within 24 h. METHODS: Patients older than 18 years undergoing laparoscopic appendectomy were prospectively enrolled at two surgical departments after informed consent was obtained. Periappendicular fluid was collected prior to appendectomy and sent for cultivation. Sensitivity, specificity and positive and negative predictive values were calculated with 95% confidence intervals (CIs) using 72-hour cultivation results as the gold standard. Patients with complicated AA as determined by the surgeon, received a three-day course of oral antibiotics. Postoperative infectious complications within 30 days after surgery were registered. RESULTS: From July 2020 to January 2021, 101 patients were included. The intraoperative diagnosis was complicated AA in 34 cases. Of these patients, six (17.6%) had bacteria cultured within 24 h after surgery, leading to a sensitivity of 60% and a specificity of 100%. The positive and negative predictive values were 1.00 and 0.96, respectively. Seven patients developed a postoperative infection (five superficial wound infections and two intra-abdominal abscess). In all cases with a positive cultivation result, the intraoperative diagnosis was complicated appendicitis and a postoperative course of antibiotics prescribed. CONCLUSION: Twenty-four-hour cultivation of the peritoneal fluid in acute appendicitis is a valid indicator for peritoneal bacterial contamination. Randomized studies are necessary to determine if this approach is suitable for targeting postoperative antibiotic treatment as a means to prevent overtreatment without increasing the risk of infectious complications.

Microalgae-derived hydrogels/membranes for phosphorus removal and recovery from aquaculture tailwater: Waste utilization and phosphorus recycling.

Efficient removal and recovery of phosphorus from aquaculture tailwater is challenging due to increasing strict water environment restrictions. This study presents a sustainable approach by using microalgae-waste-derived hydrogels/membranes for phosphorus adsorption and microalgae cultivation. Waste from Euglena gracilis (or Haematococcus pluvialis), modified with magnesium, was converted into biochars (abbreviated as MEBC or MHBC). This biochars were then combined with sodium alginate to fabricate hydrogels and with polyvinyl chloride to create membranes. Due to the almost 100 % phosphorus removal of MEBC (or MHBC) biochar, the as-obtained hydrogels/membranes demonstrated excellent phosphate adsorption, reducing total phosphorus in real aquaculture tailwater from 11 mg/L to 0. Additionally, the phosphorus-saturated hydrogel served as a phosphorus source for microalgae cultivation, while the membranes facilitated microalgae harvesting with a water flux over 40 L/m2/h. This study provides an eco-friendly solution for using microalgae-waste-derived materials to effectively address phosphorus removal and recovery challenges in aquaculture tailwater.

Should cannabis self-cultivation be part of a public health‒oriented legalization policy framework?

Cannabis control policies are increasingly being liberalized, including the legalization of non-medical cannabis use and supply in multiple settings, for example in Canada, with main policy objectives focusing on improved public health. An important while contested matter has been the appropriate design of legal cannabis supply structures and sources. These, in most Americas-based legalization settings, have included provisions for (limited) 'home cultivation'. Recent data suggest that about 8% of active consumers engage in cannabis home cultivation for their own supply, while approximately 14% are exposed to it in/around their home. Home cultivation commonly exceeds legal limits and/or occurs where not allowed, and is disproportionately associated with high-frequency and/or other risk patterns of cannabis use. In addition, home cultivation may facilitate exposure or diversion of cannabis to minors, as well as pose possible environmental exposure risks especially when occurring indoors. Given its placement in private spaces, related regulations are largely shielded from enforcement. Home cultivation, therefore, bears substantive potential to circumvent or work counter to public health‒oriented legalization policy objectives. Recent assessments of health outcomes from cannabis legalization show mixed-including multiple adverse-results, implying the need for regulatory revisions towards protecting public health outcomes. Especially in settings where extensive (e.g. commercial) retail systems were established to provide regulated, legal cannabis products to consumers, it is questionable whether home cultivation overall serves primary public health‒oriented objectives; relevant data should be expanded and used to review related provisions.

RéSUMé: Les politiques de contrôle du cannabis sont de plus en plus libéralisées, y compris la légalisation de la consommation et de l'accès au cannabis à des fins non médicales dans de nombreux contextes, par exemple au Canada, dont les principaux objectifs politiques sont axés sur l'amélioration de la santé publique. Une question importante, bien que controversée, a été la conception appropriée des structures et sources d'approvisionnement légal en cannabis. Celles-ci, dans la plupart des contextes de légalisation basés en Amériques, ont inclus des dispositions pour la 'culture à domicile' (limitée). Des données récentes suggèrent qu'environ 8 % des consommateurs actifs pratiquent la culture à domicile de cannabis pour leur propre approvisionnement, tandis qu'environ 14 % y sont exposés dans/autour de leur maison. La culture à domicile dépasse généralement les limites légales et/ou a lieu là où elle n'est pas autorisée, et est associée de manière disproportionnée à une consommation de cannabis à fréquence élevée et/ou à d'autres risques. En outre, la culture à domicile peut faciliter l'exposition ou le détournement du cannabis pour les mineurs, ainsi que présenter des risques d'exposition environnementale, en particulier lorsqu'elle se produit à l'intérieur. Étant donné qu'elles touchent à des espaces privés, les réglementations connexes sont largement à l'abri de toute application. La culture à domicile présente donc un potentiel important de contournement ou d'obstacles aux objectifs politiques axés sur la santé publique. Des évaluations récentes des conséquences sur la santé de la légalisation du cannabis montrent des résultats mitigés, y compris de multiples effets négatifs, ce qui implique la nécessité de révisions réglementaires en vue d'améliorer les résultats en matière de santé publique. En particulier dans les contextes où de vastes systèmes de vente au détail (par exemple commerciaux) ont été établis pour fournir aux consommateurs des produits à base de cannabis légaux et réglementés, on peut se demander si la culture à domicile sert globalement les principaux objectifs de santé publique; les données pertinentes devraient être élargies et utilisées pour réexaminer les dispositions y étant reliées.

Enrichment of Polycyclic Aromatic Hydrocarbon (PAH)-Degrading Strictly Anaerobic Sulfate-Reducing Cultures from Contaminated Soil and Sediment.

Sulfate-reducing bacteria (SRB) are crucial players in global biogeochemical cycling and some have been implicated in the anaerobic biodegradation of organic pollutants, including recalcitrant and hazardous polycyclic aromatic hydrocarbons (PAHs). Obtaining PAH-degrading SRB cultures for laboratories is of paramount importance in the development of the young field of anaerobic biodegradation of PAHs. SRB grow exceptionally slowly on PAH substrates and are highly sensitive to oxygen. Consequently, enrichment and maintenance of PAH-degrading SRB cultures and characterization of the biodegradation process remain a tedious and formidable task, especially for new researchers. To address these technical constraints, we have developed robust and effective protocols for obtaining and characterizing PAH-degrading SRB cultures. In this set of protocols, we describe step-by-step procedures for preparing inocula from contaminated soil or sediment, preparing anoxic medium, establishing enrichment cultures with PAHs as substrates under completely anaerobic sulfate-reducing conditions, successive culture transfers to obtain highly enriched cultures, rapid verification of the viability of SRB in slow-growing cultures, assessment of PAH degradation by extracting residuals using organic solvent and subsequent analysis by gas chromatography-mass spectrometry, and spectrophotometric determination of sulfate and sulfide in miniaturized, medium-throughput format. These protocols are expected to serve as a comprehensive manual for obtaining and characterizing PAH-degrading sulfate-reducing cultures. © 2024 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Obtaining PAH-degrading strictly anaerobic sulfate-reducing enrichment cultures from contaminated soil and sediment Support Protocol 1: Operation and maintenance of an anaerobic workstation Support Protocol 2: Setup of gas purging systems for preparing anoxic solutions Support Protocol 3: Verification of viability in slow-growing SRB enrichment cultures Support Protocol 4: Extraction of genomic DNA from low-biomass cultures Basic Protocol 2: Extraction of residual PAH from liquid culture and analysis by GC-MS Basic Protocol 3: Spectrophotometric determination of sulfate concentration in SRB cultures Basic Protocol 4: Spectrophotometric determination of sulfide concentrations in SRB cultures by the methylene blue method Alternate Protocol: Spectrophotometric determination of sulfide concentrations in SRB cultures by the colloidal copper sulfide method.

Justifying the prediction of major soil nutrients levels (N, P, and K) in cabbage cultivation.

In a recent paper by Sajindra et al. [1], the soil nutrient levels, specifically nitrogen, phosphorus, and potassium, in organic cabbage cultivation were predicted using a deep learning model. This model was designed with a total of four hidden layers, excluding the input and output layers, with each hidden layer meticulously crafted to contain ten nodes. The selection of the tangent sigmoid transfer function as the optimal activation function for the dataset was based on considerations such as the coefficient of correlation, mean squared error, and the accuracy of the predicted results. Throughout this study, the objective is to justify the tangent sigmoid transfer function and provide mathematical justification for the obtained results.•This paper presents the comprehensive methodology for the development of deep neural network for predict the soil nutrient levels.•Tangent Sigmoid transfer function usage is justified in predictions.•Methodology can be adapted to any similar real-world scenarios.

Cultivation of earthworms and analysis of associated bacterial communities during earthworms' growth using two types of agricultural wastes.

Earthworm cultivation can effectively promote the resource utilization of agricultural waste. The efficient utilization of agricultural waste by earthworms mainly depends on the microbial communities in the guts. This study used silkworm excrement and cow manure as substrates for earthworm cultivation and investigated the associated bacterial communities during earthworms' growth. The survival rate of earthworms remained above 89% after 21 days of feeding with the two substrates. Proteobacteria, Actinobacteria, and Firmicutes constituted the predominant bacterial communities in earthworm growth, accounting for over 81% of the relative abundance in both guts and vermicompost. The bacteria richness and diversity in the foregut and midgut of earthworm were lower than those in the hindgut. The prediction function of intestinal bacterial communities of earthworms cultured with two substrates mainly involved biosynthesis, decomposition and energy production.

How does a Saccharomyces cerevisiae extract influence the components of isolated rotavirus particles from stool samples collected in a clinical setting from children?

Human Rotavirus (HRV) is the causative pathogen of severe acute enteric infections that cause mortality among children worldwide. This study focuses on developing a new and effective treatment for rotavirus infection using an extract from Saccharomyces cerevisiae, aiming to make this treatment easily accessible to everyone. 15 antigens and 26 antibodies were detected in serum and stool using ELISA. The titers of HRVq1, HRVq2, HRVC1, and HRVC2 on Vero cells were determined to be 1.2x106, 3.0x106, 4.2x106, and 7.5x105 (Plaque forming unit, PFU/ml) four days after infection, respectively. The HRVq1 isolate induced cytopathic effects, i.e., forming multinucleated, rounded, enlarged, and expanding gigantic cells. RT-PCR identified this isolate, and the accession number 2691714 was assigned to GeneBank. The molecular docking analysis revealed that nonstructural proteins (NSPs) NSP1, NSP2, NSP3, NSP4, NSP5, and NSP6 exhibited significant binding with RNA. NSP2 demonstrated the highest binding affinity and the lowest binding energy (-8.9 kcal/mol). This affinity was maintained via hydrophobic interactions and hydrogen bonds spanning in length from 1.12 Å to 3.11 Å. The ADMET and bioactivity predictions indicated that the yeast extract possessed ideal solubility, was nontoxic, and did not cause cancer. The inhibitory constant values predicted for the S. cerevisiae extract in the presence of HRV vital proteins varied from 5.32 to 7.45 mM, indicating its potential as a viable drug candidate. Saccharomyces cerevisiae extract could be utilized as a dietary supplement to combat HRV as an alternative dietary supplement.

Cropping with Vicia villosa and native grass improves soil's bacterial structure and ecological network in a jujube orchard.

In a jujube orchard, cropping withgrass may influence bacterial diversity and ecological networks due to changes of physicochemical properties in soil, which has a serious effect on the stability of soil ecosystems. The aim of this study was to analyze the effects of different cultivation methods (CK: cleaning tillage; NG: cropping with native grass; VV: cropping with Vicia villosa) on the soil's bacterial structure and its co-occurrence network in a jujube orchard. The results showed that the highest moisture content, total nitrogen, and organic matter in the rhizosphere soil of a jujube orchard was found in the VV group. The soil's moisture content, total nitrogen, and organic matter in the VV group were 2.66%, 0.87 g kg-1, and 5.55 mg kg-1 higher than that found in the CK group. Compared to the CK group, the number of unique species in the rhizosphere soil in the NG and the VV groups increased by 7.33% and 21.44%. The PICRUSt and FAPROTAX analysis showed that sown grass had a greater influence on the ecological function of the soil's bacteria. Cropping with Vicia villosa and native grass significantly increased aerobic chemoheterotrophy, nitrogen respiration, nitrate reduction related to biochemical cycles, and the relative abundance of genes related to carbohydrate metabolism and the biodegradation of xenobiotics. The bacterial network complexity in the NG group was higher than that in the CK and VV groups and was greatest in the hub nodes (OTU42, Bacteroidota; OTU541, Nitrospiraceae). In this study, the ecological benefit seen in the soil's microbial function provides support to the theory that cropping with grass (Vicia villosa) increases the sustainable development of a jujube orchard.

Aster × chusanensis Growth and Phenolic Acid Composition under Different Cultivation Temperatures.

Plants of the Asteraceae family have been cultivated worldwide for economic, medicinal, and ornamental purposes, including genera such as Aster, Helianthus, and Cosmos. Numerous studies examined their secondary metabolites; however, those of Aster × chusanensis, which is a natural hybrid species in South Korea, are unclear, and optimized propagation methods should be identified. We analyzed phenolic acid concentrations in each part of Aster × chusanensis through HPLC. Further, we investigated the growth characteristics and secondary metabolite concentrations under various growth temperatures using division propagation, followed by growing at 20, 25, and 30 °C in a growth chamber. Chlorogenic acid was the primary compound, which was particularly high in the leaves. The growth characteristics did not differ significantly between temperatures, and 30 °C was most efficient for phenolic acid biosynthesis. Our results provide valuable information on optimized propagation and secondary metabolite concentrations under different temperatures of Aster × chusanensis.

Advances of 3D Cell Co-Culture Technology Based on Microfluidic Chips.

Cell co-culture technology aims to study the communication mechanism between cells and to better reveal the interactions and regulatory mechanisms involved in processes such as cell growth, differentiation, apoptosis, and other cellular activities. This is achieved by simulating the complex organismic environment. Such studies are of great significance for understanding the physiological and pathological processes of multicellular organisms. As an emerging cell cultivation technology, 3D cell co-culture technology, based on microfluidic chips, can efficiently, rapidly, and accurately achieve cell co-culture. This is accomplished by leveraging the unique microchannel structures and flow characteristics of microfluidic chips. The technology can simulate the native microenvironment of cell growth, providing a new technical platform for studying intercellular communication. It has been widely used in the research of oncology, immunology, neuroscience, and other fields. In this review, we summarize and provide insights into the design of cell co-culture systems on microfluidic chips, the detection methods employed in co-culture systems, and the applications of these models.

Development of a defined medium for the heterotrophic cultivation of Metallosphaera sedula.

The heterotrophic cultivation of extremophilic archaea still heavily relies on complex media. However, complex media are associated with unknown composition, high batch-to-batch variability, potential inhibiting and interfering components, as well as regulatory challenges, hampering advancements of extremophilic archaea in genetic engineering and bioprocessing. For Metallosphaera sedula, a widely studied organism for biomining and bioremediation and a potential production host for archaeal ether lipids, efforts to find defined cultivation conditions have still been unsuccessful. This study describes the development of a novel chemically defined growth medium for M. sedula. Initial experiments with commonly used complex casein-derived media sources deciphered Casamino Acids as the most suitable foundation for further development. The imitation of the amino acid composition of Casamino Acids in basal Brock medium delivered the first chemically defined medium. We could further simplify the medium to 5 amino acids based on the respective specific substrate uptake rates. This first defined cultivation medium for M. sedula allows advanced genetic engineering and more controlled bioprocess development approaches for this highly interesting archaeon.

Large-Scale Expansion of Human Liver Stem Cells Using Two Different Bioreactor Systems.

The assessment of human liver stem cells (HLSCs) as cell therapeutics requires scalable, controlled expansion processes. We first focused on defining appropriate process parameters for HLSC expansion such as seeding density, use of antibiotics, optimal cell age and critical metabolite concentrations in conventional 2D culture systems. For scale-up, we transferred HLSC expansion to multi-plate and stirred-tank bioreactor systems to determine their limitations. A seeding density of 4000 cells cm-2 was needed for efficient expansion. Although growth was not significantly affected by antibiotics, the concentrations of lactate and ammonia were important. A maximum expansion capacity of at least 20 cumulative population doublings (cPDs) was observed, confirming HLSC growth, identity and functionality. For the expansion of HLSCs in the multi-plate bioreactor system Xpansion (XPN), the oxygen supply strategy was optimized due to a low kLa of 0.076 h-1. The XPN bioreactor yielded a final mean cell density of 94 ± 8 × 103 cells cm-2, more than double that of the standard process in T-flasks. However, in the larger XPN50 device, HLSC density reached only 28 ± 0.9 × 103 cells cm-2, while the glucose consumption rate increased 8-fold. In a fully-controlled 2 L stirred-tank bioreactor (STR), HLSCs expanded at a comparable rate to the T-flask and XPN50 processes in a homogeneous microenvironment using advanced process analytical technology. Ultimately, the scale-up of HLSCs was successful using two different bioreactor systems, resulting in sufficient numbers of viable, functional and undifferentiated HLSCs for therapeutic applications.