Medium Formulation Research Articles

Shifting job expectations in the era of generative AI hype – perspectives of journalists and copywriters

PurposeThis interview study examines Hungarian journalists' and copywriters' expectations of generative AI’s impact on their professions and factors influencing these views during a period of hype.Design/methodology/approachWhile acknowledging the specialized knowledge of journalists and copywriters relative to the general public, the study employs the sociology of expectations framework to interpret their anticipations not as objective forecasts of the future, but rather as phenomena shaped by diverse influences. The research comprises 30 semi-structured interviews conducted in spring 2023 to explore these expectations and their contributing factors.FindingsResults reveal ChatGPT’s media coverage as pivotal, encouraging the professionals interviewed to experiment with AI, reassess their roles, and cause a shift in their job expectations. At the same time, this shift was limited. Skepticism about hyperbolic media formulations, their own experiences with ChatGPT and projecting its constraints into the future, contextual factors, and optimism bias contributed to moderating their expectations. They perceived AI as an enhancer of efficiency and quality, not as a radical disruptor. Copywriters were more open to integrating AI in their work, than journalists.Research limitations/implicationsThe results underscore the importance of further research to explore subjective experiences associated with technological change, particularly considering their complex social, psychological, and cultural influences.Originality/valueThe study uniquely contributes to the sociology of expectations by highlighting how a complex interplay of factors can shape professionals' anticipation of the impact of AI on their careers, including optimism bias and media hype.

β-glycerophosphate, not low magnitude fluid shear stress, increases osteocytogenesis in the osteoblast-to-osteocyte cell line IDG-SW3

ABSTRACT Aim As osteoblasts deposit a mineralized collagen network, a subpopulation of these cells differentiates into osteocytes. Biochemical and mechanical stimuli, particularly fluid shear stress (FSS), are thought to regulate this, but their relative influence remains unclear. Here, we assess both biochemical and mechanical stimuli on long-term bone formation and osteocytogenesis using the osteoblast-osteocyte cell line IDG-SW3. Methods Due to the relative novelty and uncommon culture conditions of IDG-SW3 versus other osteoblast-lineage cell lines, effects of temperature and media formulation on matrix deposition and osteocytogenesis were initially characterized. Subsequently, the relative influence of biochemical (β-glycerophosphate (βGP) and ascorbic acid 2-phosphate (AA2P)) and mechanical stimulation on osteocytogenesis was compared, with intermittent application of low magnitude FSS generated by see-saw rocker. Results βGP and AA2P supplementation were required for mineralization and osteocytogenesis, with 33°C cultures retaining a more osteoblastic phenotype and 37°C cultures undergoing significantly higher osteocytogenesis. βGP concentration positively correlated with calcium deposition, whilst AA2P stimulated alkaline phosphatase (ALP) activity and collagen deposition. We demonstrate that increasing βGP concentration also significantly enhances osteocytogenesis as quantified by the expression of green fluorescent protein linked to Dmp1. Intermittent FSS (~0.06 Pa) rocker had no effect on osteocytogenesis and matrix deposition. Conclusions This work demonstrates the suitability and ease with which IDG-SW3 can be utilized in osteocytogenesis studies. IDG-SW3 mineralization was only mediated through biochemical stimuli with no detectable effect of low magnitude FSS. Osteocytogenesis of IDG-SW3 primarily occurred in mineralized areas, further demonstrating the role mineralization of the bone extracellular matrix has in osteocyte differentiation.

Generating suspension-adapted human mesenchymal stromal cells (S-hMSCs) for the scalable manufacture of extracellular vesicles

BackgroudHuman mesenchymal stromal cells (hMSCs) are a naturally adherent cell type and one of the most studied cellular agents used in cell therapy over the last 20 years. Their mechanism of action has been primarily associated with paracrine signaling, which has contributed to an increase in the number of studies focused on hMSC-related extracellular vesicles (EVs). MethodsIn this study, we demonstrate for the first time that human telomerase reverse transcriptase (hTERT) immortalized hMSCs can be adapted to suspension culture, eliminating the need for microcarriers or other matrixes to support cell growth. ResultsThis novel cell line, named suspension hMSCs (S-hMSCs), has a doubling time of approximately 55 hours, with a growth rate of 0.423/d. Regarding its immunophenotype characteristics, S-hMSCs retained close to 90% of CD73 and CD105 expression levels, with the CD90 receptor being downregulated during the adherent to suspension adaptation process. An RNA sequencing analysis showed an upregulation of the transcripts coding for CD44, CD46 and CD47 compared to the expression levels in AT-hMSCs and hTERT-hMSCs. The cell line herein established was able to generate EVs using a chemically defined medium formulation with these nanoparticles averaging 150 nm in size and displaying the markers CD63, CD81, and TSG101, while not expressing the negative marker calnexin. ConclusionThis body of evidence, combined with the visual confirmation of EV presence using transmission electron microscopy, demonstrates the EV-producing capabilities of the novel S-hMSCs. This cell line provides a platform for process development, drug discovery and translational studies in the EV field.

Evaluation of 21 different media on shoot regeneration in 11 cultivars of citrus using juvenile tissue

Citrus is one of the most important global fruit crops, with oranges accounting for more than half the total production. Citrus can be genetically engineered for improved traits, but the process is severely limited by shoot regeneration rates, especially in commercial varieties. Although standardizing a transformation protocol is difficult due to genotype and season dependencies, use of an optimal mineral nutrient basal culture medium increases the success of recovering transgenic Citrus cells. The purpose of this study is to improve shoot regeneration from juvenile tissue in multiple Citrus genotypes. Explants of 11 Citrus cultivars were regenerated in 21 published media to determine the optimal media for each genotype. The number of shoots from juvenile tissue were counted 90 days after the explants were first cultured. The type of basal medium strongly affected the rates of shoot regeneration from Citrus juvenile tissue, and the effectiveness was negatively correlated with the level of ammonium. For each Citrus grouping, optimized media formulations were determined, and potential improvements were predicted. In general, medium R7100 appeared to be the most effective for regeneration of the Citrus genotypes tested.

A Method to Evaluate Forchheimer Resistance Coefficients for Permeable Screens and Air Louvers Modelled as a Porous Medium

Porous medium models are commonly used in Computational Fluid Dynamics (CFD) to simulate flow through permeable screens of various types. However, the setup of these models is often limited to replicating a pressure drop in cases where fluid inflow is orthogonal to the screen. In this work, a porous medium formulation that employs a non-diagonal Forchheimer tensor is presented. This formulation is capable of reproducing both the pressure drop and flow deflection under varying inflow angles for complex screen geometries. A general method to determine the porous model coefficients valid for both diagonal and non-diagonal Forchheimer tensors is proposed. The coefficients are calculated using a nonlinear least-squares optimisation based on an analytical solution of a special case of the Navier–Stokes equations. The applicability of the proposed method is evaluated in four different scenarios supplemented by local CFD simulations of permeable screens: wire mesh, perforated screens, air louvers, and expanded mesh panels. The practical application of this method is demonstrated in the modelling of windbreaks and permeable double-skin facades, which typically employ the aforementioned types of porous screens.

Elevated Lipid Metabolites in Stored Clinical OCA Media Correlate With Chondrocyte Death

Background: A major limitation of osteochondral allografts (OCA) is the deterioration of cartilage health associated with cell death during prolonged storage. However, little is known about the mechanisms that contribute to chondrocyte death during storage. Purpose/Hypothesis: This study aimed to determine whether bioactive lipid metabolites accumulate in the storage media of OCA and whether they are associated with a loss of chondrocyte viability during prolonged storage. It was hypothesized that free fatty acids (FFAs) would accumulate over time in the storage media of OCA and adversely affect cartilage health during storage. Study Design: Controlled laboratory study. Methods: A group of 21 (n = 6-8 OCA/treatment group) fresh human hemicondylar OCA tissues and media were analyzed after 7, 28, and 68 days of prolonged cold (4°C) storage. Targeted mass spectrometry analysis was used to quantify bioactive FFAs, as well as primary (lipid hydroperoxide [ROOH]) and secondary (malondialdehyde) lipid oxidation products. Chondrocyte viability was measured using a fluorescence-based live/dead assay and confocal microscopy. Results: The concentration of all targeted fatty acid metabolites in storage media was significantly increased with increased cold storage time (P < .05). ROOH was significantly higher on day 28 of cold storage. No difference in secondary ROOH products in storage media was observed. Chondrocyte viability significantly declined in both the en face and the vertical cross-sectional analysis with increased cold storage time and inversely correlated with fatty acid metabolites (P < .05). Conclusion: It is well established that elevated levels of certain FFAs and lipid oxidation products can alter cell function and cause cell death via lipotoxicity and other mechanisms. This work is the first to identify elevated levels of FFA metabolites and primary oxidation lipid products in the storage media from clinical OCA. The concentrations of FFA metabolites were measured at levels (>100 µM) known to induce cell death and were directly correlated with chondrocyte viability. Clinical Relevance: These findings provide important targets for understanding why cartilage health declines during cold storage, which can be used to optimize media formulations and improve graft health.

Formulation of Protein Rich Media and Broth Using Chicken Feather

Formulation of Protein Rich Media and Broth Using Chicken Feather

High coverage of targeted lipidomics revealed lipid changes in the follicular fluid of patients with insulin-resistant polycystic ovary syndrome and a positive correlation between plasmalogens and oocyte quality.

Polycystic ovary syndrome with insulin resistance (PCOS-IR) is the most common endocrine and metabolic disease in women of reproductive age, and low fertility in PCOS patients may be associated with oocyte quality; however, the molecular mechanism through which PCOS-IR affects oocyte quality remains unknown. A total of 22 women with PCOS-IR and 23 women without polycystic ovary syndrome (control) who underwent in vitro fertilization and embryo transfer were recruited, and clinical information pertaining to oocyte quality was analyzed. Lipid components of follicular fluid (FF) were detected using high-coverage targeted lipidomics, which identified 344 lipid species belonging to 19 lipid classes. The exact lipid species associated with oocyte quality were identified. The number (rate) of two pronuclear (2PN) zygotes, the number (rate) of 2PN cleaved embryos, and the number of high-quality embryos were significantly lower in the PCOS-IR group. A total of 19 individual lipid classes and 344 lipid species were identified and quantified. The concentrations of the 19 lipid species in the normal follicular fluid (control) ranged between 10-3 mol/L and 10-9 mol/L. In addition, 39 lipid species were significantly reduced in the PCOS-IR group, among which plasmalogens were positively correlated with oocyte quality. This study measured the levels of various lipids in follicular fluid, identified a significantly altered lipid profile in the FF of PCOS-IR patients, and established a correlation between poor oocyte quality and plasmalogens in PCOS-IR patients. These findings have contributed to the development of plasmalogen replacement therapy to enhance oocyte quality and have improved culture medium formulations for oocyte in vitro maturation (IVM).

OPTIMIZATION OF α-AMYLASE SYNTHESIS BY BACILLUS VELEZENSIS USING TAGUCHI EXPERIMENTAL DESIGN

This research employs the Taguchi experimental design to optimize the production of α-amylase, a versatile enzyme with diverse industrial applications, using Bacillus velezensis. Thirteen key parameters, including pH, temperature, agitation, inoculum size, aeration and various carbon and nitrogen sources, were systematically investigated using the Taguchi L27 (313) orthogonal array. Initial screening of six seed media formulations identified SM6 (0.5% peptone, 0.5% yeast extract, 0.1% K2 HPO4 ) as the optimal medium, producing 2.8 U mL-1 of α-amylase. Further optimization under the Taguchi framework yielded a maximum α-amylase activity of 1097.31 U mL-1 and total protein of 1230 mg mL-1 at pH 5, 34°C, 4% moong husk as carbon source, and 2% soybean cake as nitrogen source. Two-factor interaction analysis revealed that a temperature of 34°C with 2% soybean cake, and 4% moong husk with 2% soybean cake, provided the best results. Validation experiments confirmed a 31.2% enhancement in α-amylase production under the optimized conditions. This study highlights the efficacy of the Taguchi design in systematically optimizing complex bioprocesses, paving the way for improved α-amylase synthesis with B. velezensis.

Enhancing recombinant antibody yield in Chinese hamster ovary cells.

A range of recombinant monoclonal antibodies (rMAbs) have found application in treating diverse diseases, spanning various cancers and immune system disorders. Chinese hamster ovary (CHO) cells have emerged as the predominant choice for producing these rMAbs due to their robustness, ease of transfection, and capacity for posttranslational modifications akin to those in human cells. Transient transfection and/or stable expression could be conducted to express rMAbs in CHO cells. To bolster the yield of rMAbs in CHO cells, a multitude of approaches have been developed, encompassing vector optimization, medium formulation, cultivation parameters, and cell engineering. This review succinctly outlines these methodologies when also addressing challenges encountered in the production process, such as issues with aggregation and fucosylation.

The influence of the nutrient composition medium on the intensity micropropagation in vitro Fragaria × Ananassa duch.

Strawberry is one of the most economically significant crops in the world berry growing. In the course of research, the influence of the mineral composition of nutrient media on the growth and development of strawberry plants at the stage of micro-propagation proper was studied for further optimization of the technology of microclonal reproduction. The purpose of the work is to select the optimal nutrient medium for micropropagation of strawberry in vitro in the production system of healthy planting material. The research was carried out in the laboratory of biotechnology of the Russian research Institute of Fruit Grop Breeding according to generally accepted methods. The objects were important commercial varieties of garden strawberries (Fragaria × ananassa Duch.) of foreign breeding: Alba (NF 311), Darselect, Kimberly, Clery, Syria (NF 137), Florence. Variants of the nutrient media formulations used: Lee and de Fossard, Gamborg and Eveleigh (B5), Knop, Murashige-Skoog with the addition of 0.8 mg/ l cytokinin 6-BAP. Optimal recipes of the nutrient medium that promote intensive proliferation and root formation in plants have been determined. For the cultivation of the studied strawberry varieties at the stage of actual micro-propagation with a high degree of regeneration, the optimal nutrient medium is the MS medium. On the nutrient Knop’s medium, the micro-plants on the second passage acquired a reddish color, not characteristic of the studied varieties. The maximum values of plant height were noted in the control variant with MS nutrient medium in the varieties Darselect (9.1 mm) and Kimberly (8.6 mm). In all other variants of the studied nutrient media, the average height of plants did not exceed 7.7 mm. Media according to the LF and B5 recipe can be used to obtain microplants suitable for planting in ex vitro conditions, excluding the rooting stage, to accelerate the production of healthy planting material. A different reaction of strawberry varieties to the mineral composition of nutrient media has been established.

Nutrient optimization in bioleaching: are we overdosing?

The general trend in biomining (i.e., bioleaching and biooxidation) is the use of media with high concentrations of the nutrients (nitrogen as ammonium, phosphorous as phosphate, and K), which are considered to be essential for microbial growth. The depletion of any of the nutrients would affect negatively the bioleaching (and biooxidation) capacity of the microorganisms, so the formulation of the different media ensures that there is a surplus of nutrients. However, some of these nutrients (e.g., phosphate, K) may be already present in the ore and are made available to the microorganisms when the ore is exposed to the low-pH media used during bioleaching. The effect of phosphate addition (109 mg/L) and depletion on the bioleaching of low-grade sulfidic ore alongside the determination of ammonium (i.e., 25 mg/L, 50 mg/L, 109 mg/L, 409 mg/L, and 874 g/L) requirements were studied. The results of the experiments presented showed that the addition of phosphate did not have any effect on the bioleaching of the low-grade sulfidic ore while the addition of ammonium was necessary to obtain higher redox potentials (>650 mV vs. Ag/AgCl) and higher metal (Co, Cu, Ni, and Zn) dissolutions. Temperature was the factor that shaped the microbial communities, at 30°C, the microbial community at the end of all the experiments was dominated by Acidithiobacillus sp. as well as at 42°C, except when nutrients were not added and Sulfobacillus sp. was the dominant microorganism. At 55°C, DNA recovery was unsuccessful, and at 60°C, the microbial communities were dominated by Sulfolobus sp. In conclusion, the amount of nutrients in bioleaching could be reduced significantly to achieve the redox potentials and metal dissolution desired in bioleaching without affecting the microbial communities and bioleaching efficiencies.

Overcoming Microbial Inhibition of S. Sonnei Through the Exploitation of Genomically Predicted Antibiotic Resistance Profiles for the Development of Food Enrichment Media

Linking outbreaks of Shigella spp. to specific foods is challenging due to poor selectivity of current enrichment media. We have previously shown that enrichment media, tailored to the genomically-predicted antimicrobial resistance (AMR) of Shiga toxigenic E. coli strains, enhances their isolation from foods. This study investigates the application of this approach for Shigella isolation. The AMR gene profiles of 21,908 published S. sonnei genomes indicated a high prevalence of genes conferring resistance to streptomycin (aadA, aph(3″)-Ib, aph(6)-Id, 92.8%), sulfonamides (sul1, sul2, 74.8%), and/or trimethoprim (dfrA, 96.2%). Genomic analysis and antibiotic susceptibility testing conducted with a panel of 17 outbreak-associated S. sonnei strains confirmed the correlation of AMR gene detection with resistance phenotypes. Supplementation of Shigella Broth (SB) with up to 400 µg/mL of trimethoprim or sulfadiazine did not suppress the growth of sensitive strains, whereas 100 µg/mL of streptomycin increased the selectivity of this broth. All three antibiotics increased the selectivity of modified Tryptone Soya Broth (mTSB). Based on these results, supplemented media formulations were developed and assessed by measuring the relative growth of S. sonnei in cultures coinoculated with a strain of bacteriocin-producing E. coli that is inhibitory to Shigella growth. S. sonnei was not recovered from cocultures grown in SB or mTSB without antibiotics. In contrast, media supplemented with streptomycin at 50 and 100 µg/mL, trimethoprim at 25 and 50 µg/mL, and sulfadiazine at 100 µg/mL increased the relative proportion of S. sonnei in postenrichment cultures. The enhanced recovery of resistant S. sonnei strains achieved in this study indicates that, in cases where genomic data are available for clinical S. sonnei isolates, customization of selective enrichment media based on AMR gene detection could be a valuable tool for supporting the investigation of foodborne shigellosis outbreaks.

Novel cereal bran based low-cost liquid medium for enhanced growth, multifunctional traits and shelf life of consortium biofertilizer containing Azotobacter chroococcum, Bacillus subtilis and Pseudomonas sp.

The present study was carried out to valorise cereal (rice and wheat) bran for the development of low-cost liquid consortium bioformulation. Different concentrations of bran-based liquid media formulations were evaluated for the growth of consortium biofertilizer cultures (Azotobacter chroococcum, Bacillus subtilis and Pseudomonas sp.). Among the bran-based formulations, wheat bran-based formulation WB5, exhibited the highest viable cell of 10.68 ± 0.09 Log10 CFU/ml and 12.63 ± 0.04 Log10 CFU/ml for Azotobacter chroococcum and Bacillus subtilis whereas for Pseudomonas sp., rice bran based bioformulation RB5 recorded maximum viability (12.71 ± 0.05 Log10 CFU/ml) after 72 h of incubation. RB51RB1: 1:10 (Rice bran: distilled water)RB2: 1:10 (Rice bran: distilled water)RB3: 1:10 (Rice bran: distilled water)RB4: 1:10 (Rice bran: distilled water)RB5: 5:10 (Rice bran: distilled water)1 and WB52WB1: 1:10 (wheat bran: distilled water)WB2: 2:10 (wheat bran: distilled water)WB3: 3:10 (wheat bran: distilled water)WB4: 4:10 (wheat bran: distilled water)WB5: 5:10 (wheat bran: distilled water)2liquid formulations were further optimized for enhanced shelf life using 5, 10 and 15 mM of trehalose, 0.05 and 0.1% carboxymethyl cellulose, and 0.5 and 1.0% glycerol. Following the peak growth at 72 h of incubation, a gradual decrease in the viable population of consortium biofertilizer cultures was observed in all the liquid formulations. The WB5 and RB5 formulations with 15 mM trehalose and 0.1% CMC, not only recorded significantly highest cell count of consortium biofertilizer cultures, but also maximally supported multi-functional traits i.e., phosphate and zinc solubilization, ammonia and IAA production up to 150 days. Further evaluation of seedling emergence and growth of wheat (PBW 826) under axenic conditions recorded WB5 amended with 15 mM trehalose-based consortium bioformulation to exhibit maximum emergence and growth of wheat seedlings. This low-cost liquid formulation can be used for large-scale biofertilizer production as a cost-effective liquid biofertilizer production technology.

Optimized quantification of fatty acids in complex media using advanced analytical techniques.

Various medium formulations contain essential fatty acids at concentrations ranging from 10 to 100 mg/L. Accurate and precise lipid measurement in media is crucial for monitoring media quality and conducting studies on lipids in the context of cell culture. This study employed two-dimensional gas chromatography (GC × GC) analyses to offer enhanced resolution, sensitivity, and separation performance compared to GC. Quantification of fatty acid methyl esters (FAMEs) in a medium was conducted using GC × GC combined with a high-resolution mass spectrometer and flame ionization detector, considering potential interference from nonionic surfactant Tween 80, which was precipitated and removed by optimizing the concentration of cobalt thiocyanate (CTA) solution during pretreatment. This advanced analytical approach enabled identification of cis and trans isomers of identical molecular weights and determination of the location and number of double bonds in the same carbon number structure. Our analysis identified 36 FAMEs within the C6-C24 region, and a 5% CTA solution was optimal for efficient removal of Tween 80 during lipid extraction. Additionally, this advanced method minimized FAME contamination and loss during pretreatment, thereby significantly reducing the sample volume required to detect trace levels of FAMEs. This improvement led to a fatty acid recovery rate of 106% while maintaining the average relative standard deviation for the target FAMEs of about 3%. Our research paves the way for future investigation into medium quality control and the role of fatty acids in cell culture. This offers the possibility for economical and effective trace quantification of fatty acids in complex media.

Optimization of Fermentation Culture Medium for Sanghuangporus alpinus Using Response-Surface Methodology

The newly identified Sanghuangporus alpinus species of the Sanghuang mushroom genus has been found to possess significant medical benefits. However, the current artificial cultivation technology has not reached the requisite maturity. The response-surface methodology (RSM) was used to optimize the Sanghuangporus alpinus culture medium formulation and evaluate the functional activity of S. alpinus exopolysaccharides. First, a single-factor experiment was conducted to screen for optimal carbon and nitrogen sources for S. alpinus. Then, using Box–Behnken’s central composite design, a response-surface experiment was conducted to determine optimal culture parameters. Finally, the rationality of those parameters was assessed in a shaking flask experiment. The optimal culture parameters, determined through regression analysis, were 20.20 ± 0.17 g/L fructose (carbon source), 7.29 ± 0.10 g/L yeast extract (nitrogen source), and 0.99 ± 0.01 g/L dandelion. With optimization, the S. alpinus yield increased to 12.79 ± 1.41 g/L, twice that obtained from the initial culture medium. The S. alpinus exopolysaccharide exhibited an excellent antioxidant capacity, with the strongest scavenging effect noted on ABTS free radicals (lowest half-inhibitory concentration: 0.039 mg/mL). Additionally, this exopolysaccharide effectively inhibited various cancer cells, exhibiting the strongest activity against human glioma cells U251 (half-inhibitory concentration: 0.91 mg/mL). The RSM used to optimize the fermentation culture parameters of S. alpinus significantly increased the mycelial biomass. The improvement of Sanghuangporus alpinus yield through liquid fermentation and optimizing the fermentation medium could fill the existing gap in the cultivation of Sanghuangporus alpinus, as well as provide valuable data for the large-scale production of S. alpinus.

Generation of 'semi-guided' cortical organoids with complex neural oscillations.

Temporal development of neural electrophysiology follows genetic programming, similar to cellular maturation and organization during development. The emergent properties of this electrophysiological development, namely neural oscillations, can be used to characterize brain development. Recently, we utilized the innate programming encoded in the human genome to generate functionally mature cortical organoids. In brief, stem cells are suspended in culture via continuous shaking and naturally aggregate into embryoid bodies before being exposed to media formulations for neural induction, differentiation and maturation. The specific culture format, media composition and duration of exposure to these media distinguish organoid protocols and determine whether a protocol is guided or unguided toward specific neural fate. The 'semi-guided' protocol presented here has shorter induction and differentiation steps with less-specific patterning molecules than most guided protocols but maintains the use of neurotrophic factors such as brain-derived growth factor and neurotrophin-3, unlike unguided approaches. This approach yields the cell type diversity of unguided approaches while maintaining reproducibility for disease modeling. Importantly, we characterized the electrophysiology of these organoids and found that they recapitulate the maturation of neural oscillations observed in the developing human brain, a feature not shown with other approaches. This protocol represents the potential first steps toward bridging molecular and cellular biology to human cognition, and it has already been used to discover underlying features of human brain development, evolution and neurological conditions. Experienced cell culture technicians can expect the protocol to take 1 month, with extended maturation, electrophysiology recording, and adeno-associated virus transduction procedure options.

Complex nitrogen sources from agro-industrial byproducts: impact on production, multi-stress tolerance, virulence, and quality of Beauveria bassiana blastospores.

Biological control through entomopathogenic fungi provides essential ecological services in the integrated management of agricultural pests. In the context of submerged liquid fermentation, the nutritional composition significantly influences the ecological fitness, virulence and quality of these fungi. This study specifically explores the impact of various complex organic nitrogen sources derived from agro-industrial byproducts on the submerged liquid fermentation of Beauveria bassiana, a versatile entomopathogenic fungus known for producing hydrophilic yeast-like blastospores. Notably, manipulating the nitrogen source during submerged cultivation can influence the quality, fitness, and performance of blastospores. This research identifies cottonseed flour as the optimal low-cost nitrogen source, contributing to increased production yields, enhanced multi-stress tolerance, heightened virulence with extended shelf life and long-term bioactivity. These findings deepen our understanding of the critical role of nitrogen compound selection in liquid media formulation, facilitating the production of ecologically fit and virulent blastospores for more effective pest biocontrol programs.

An approach for topology optimization of heat sinks for two-phase flow boiling: Part 2 – Model calibration and experimental validation

Flow boiling offers increased heat transfer coefficients and effective heat capacity rates that can be leveraged by topology optimization (TO) to generate high-performance microchannel heat sinks. Topology optimization of heat sinks having two-phase flows can be accomplished using the homogenization approach wherein designs are represented as spatially varying distributions of microstructures, as demonstrated in Part 1 of this two-part study. Flow and heat transfer within the microstructures are captured using a two-phase mixture model featuring an effective porous medium formulation. However, closure of these governing equations requires empirical correlations for pressure drop and heat transfer that are specific to the operating conditions, geometry, and surface finish. Therefore, it must be demonstrated these available correlations can be successfully calibrated over a range of microstructural variations present within the homogenization framework, so as to attain the required prediction generality and accuracy needed to ensure the resulting designs achieve Pareto-optimality. This Part 2 of our study successfully demonstrates a comprehensive end-to-end process for two-phase flow model calibration, topology optimized design generation, and experimental validation of optimized performance for flow boiling heat sinks. To this end, a TO algorithm with the homogenization approach and a two-phase mixture model is implemented using available flow boiling correlations for microchannels. A set of uniform pin fin calibration samples are additively manufactured and experimentally tested under flow boiling at various flow rates and heat inputs for model calibration. All of the unknown/free coefficients in the adopted correlations are determined by minimizing the error between the model predictions and the experimental measurements using gradient-based optimization. The calibrated topology optimization algorithm is then used to generate a Pareto-optimal set of heat sinks optimized for minimum pressure drop and thermal resistance during flow boiling. Experimental characterization of these additively manufactured heat sinks, unseen during the model coefficient calibration process, reveals that the measured Pareto optimality curve matches that predicted by the topology optimization algorithm. Lastly, a heat sink design is generated for a design space involving multiple hot spots and background heating to showcase the capability of the experimentally calibrated two-phase topology optimization algorithm at handling complex boundary conditions. The optimized heat sink intelligently distributes an adequate amount of coolant flow to each of the heated regions to avoid local dry-out. This work demonstrates a complete framework for two-phase topology optimization of heat sinks through experimental calibration of flow boiling correlations to the porous medium used by the homogenization approach.

Growth and organotypic branching of lung-specific microvascular cells on 2D and in 3D lung-derived matrices

Tissue-specific endothelial cells have vital roles in maintenance and functioning of native tissues with constant reciprocal crosstalk with resident cells. Three-dimensional (3D) physio-mimetic in vitro models which incorporate lung-specific microvasculature are needed to model lung-related diseases which involve modulation of endothelial cell behavior like cancer. In this study, we investigated the growth kinetics, morphological changes and responses to biological cues of lung microvasculature on two-dimensional (2D) and in lung matrix-derived 3D hydrogels. HUVEC and HULEC-5a cells were cultured on 2D and compared for their growth, morphologies, and responses to varying growth medium formulations. Brightfield and immunofluorescence imaging was performed to assess differences in morphology. For 3D cultures, native bovine lungs were decellularized, lyophilized, solubilized, and reconstituted into hydrogel form in which endothelial cells were embedded. Cell growth and organotypic branching was monitored in 3D hydrogels in the presence of varying biological cues including lung cancer cell secretome. HUVEC and HULEC-5a cells demonstrated comparable growth and morphology on 2D. However, in 3D lung-derived ECM hydrogels, tissue-specific HULEC-5a cells exhibited much better adaptation to their microenvironment, characterized by enhanced organotypic branching and longer branches. HULEC-5a growth was responsive to lung cancer cell-conditioned medium in both 2D and 3D conditions. In 3D, the concentration of ECM ligand significantly affected cell growth in long-term culture where molecular crowding had an inhibitory role. Our data reveals that HULEC-5a cells offer a reliable alternative to frequently pursued HUVECs with comparable growth and morphology. Due to their intrinsic program for cellular crosstalk with resident cells, the use of tissue-specific endothelium constitutes a vital aspect for modeling physiological and pathological processes. Furthermore, our study is the first demonstration of the synergy between lung-specific microvasculature with lung-specific ECM within a 3D in vitro model.