Enabling supratheoretical isopropanol yields from carbon-negative glucose fermentations with a Clostridium acetobutylicum- Clostridium ljungdahlii coculture.

Glioblastoma (GBM) is the most common and lethal primary brain tumor, originating from glial cells and classified as grade IV by the World Health Organization (WHO). Standard treatment includes resection, radiotherapy, and the use of temozolomide (TMZ), a DNA-alkylating agent; however, therapeutic success is limited due to tumor heterogeneity and resistance mechanisms. Selenium (Se) is an essential element involved in selenoprotein formation, with reported anti-inflammatory, antioxidant, and antitumor activities. Among organoselenium compounds (OrganoSe), Diphenyl Diselenide (DPDS) has demonstrated antitumor, antioxidant, and anti-inflammatory properties, but its effects in human GBM are underexplored. Here, we investigated, for the first time, the antitumor capacity of DPDS, both alone and in combination with TMZ, in U87-MG cells. We evaluated cell viability, density, migration, biochemical and molecular pathways, including oxidative stress, inflammation, apoptosis, hypoxia response, epigenetic regulation, cell cycle, DNA damage response, and epithelial–mesenchymal transition (EMT). DPDS (20 µM) in combination with TMZ (100 µM) reduced cell viability and density. Co-exposure decreased cellular migration and oxidative stress markers (ROS production, SOD, and CAT activities). Gene expression analysis revealed downregulation of genes related to redox defense and inflammatory signaling, such as NRF2, KEAP1, GPX1, GPX4, TRXR1, SELENOS, SELENOK, SELENOI, NFKB1, IL6, IL-1B, TNFα, PTGS2, BAX, BAD, BCL-2, HIF1A, HDAC4, NEK1, NEK2, and SNAIL1. The results indicate that DPDS, particularly in combination with TMZ, attenuated pro-inflammatory and antioxidant profiles and interfered with pathways critical for GBM survival.

Hollow Fiber Membrane Bioreactors (HFMBs) support mammalian cell culture at high cell density by enabling effective transport of nutrients, gases and metabolites to/from cells and by offering high membrane surface area-to-bioreactor volume ratios, scalability and flexibility in design. A recent study showed that shell-and-tube HFMBs with cells in the extracapillary space (ECS) for medicine may be designed to mimic the bone architecture for bone tissue engineering (TE). A more transport-efficient HFMB, the BRx-HFMB, consists of a 3D stack of alternating cross-woven mats of microporous and gas-permeable hollow fiber membranes with cells in the ECS and medium flowing inside the membranes. This design was shown to support the culture of a large mass of densely packed cells with high oxygen and nutrient metabolic demands similar to bone cells. Its biomimicry and rationale for good performance are poorly characterized, hindering exploitation of its characteristics for bone TE. Herein, we report the architectural and biomimetic characterization of pore/void distribution in the ECS of laboratory BRx-HFMBs using non-invasive non-destructive micro-computed tomography and advanced image analysis to minimize artifacts. The results suggest that the ECS pore architecture and specific surface area of BRx-HFMBs mimic those of bone tissue, favoring cell migration around, and adhesion on, membranes at clinical cell densities. The membrane network architecture enables cell perfusion with medium, and membranes act as spatially distributed oxygen sources promoting oxygen transport through the cell construct over longer distances than in static bioreactors. This supports the use of BRx-HFMBs to develop cellular models of bone tissue for drug testing and precision medicine.

Physiological effects of zinc availability in the coral endosymbiont Cladocopium goreaui in a continuous culture system.

Multilayer microbial framework of aerobic granule microecosystems: Integrating community ecology, genetic networks, and enhancement strategies.

Effect of environmental parameters on growth and calcification in marine coccolithophore Gephyrocapsa huxleyi CCMP371.

Hydrogenophilus thermoluteolus strain TH-1 is a thermophilic hydrogen-oxidizing bacterium that can autotrophically grow using H2, CO2, and O2 with the highest rate among autotrophs. However, despite its significant potential for use in biomanufacturing from CO2, no model to date has accurately described strain TH-1 growth under both H2- and O2-limiting conditions. Here, we report on a growth model for continuous culture. Theoretical equations were established for cell yields from the substrate consumption, overall mass transfer coefficients of H2 and O2, cell densities, and dilution rates. These equations were globally fitted to a series of cell culture profiles. Global fitting showed that the model closely reproduced cell production in continuous culture under H2-limiting and O2-limiting conditions. This model will be useful for understanding culture profiles and optimizing production.

Relationship between the morphoanatomy of Juglans regia L. leaves and the intensity of bacterial disease in walnut trees.

Bupivacaine-Induced Regeneration in Rabbit Extraocular Muscles: Implications for the Treatment of Strabismus.

Anthropogenic particles and legacy pollutants combined with transcriptomic and multi-biomarker responses for an integrated assessment of the health status of mussels Mytilus galloprovincialis in two locations of the Bay of Biscay.

Methylotrophic yeasts are promising hosts for bioproduction from methanol, but their growth is significantly reduced in the presence of methanol. In this study, we investigated metabolic bottlenecks in Komagataella phaffii, Ogataea polymorpha, and Candida boidinii during methanol assimilation through comparative metabolome analysis. We found a common decrease in α-ketoglutarate-derived amino acids and tricarboxylic acid (TCA) cycle intermediates in all three species during methanol assimilation, suggesting a metabolic bottleneck around the TCA cycle entry. Additionally, K. phaffii and O. polymorpha accumulate trehalose in the presence of methanol. ΔΔG analysis suggested reduced activity of pyruvate kinase and TCA cycle entry enzymes as potential rate-limiting steps. Biosynthesis of TCA cycle-derived amino acids, including Glu, Arg, and Pro, is a limiting factor because supplementation with these amino acids significantly enhanced growth rates and final cell density for all three species. Furthermore, disruption of the trehalose biosynthesis gene (TPS2) in K. phaffii further improved growth, indicating that trehalose accumulation negatively affects methanol-based growth. Amino acid supplementation with TPS2 disruption in K. phaffii resulted in a 1.2-fold increase in the specific growth rate and a 2.73-fold increase in final cell density. These findings provide insights into the metabolic limitations of methylotrophic yeast growth in the presence of methanol and offer strategies for improving bioproduction efficiency.

Advancing bioprocess monitoring: data fusion and ANN-based prediction of arginine concentration in monoclonal antibody-producing CHO cell cultures.

This study aimed to evaluate the feasibility of visualizing and quantitatively analyzing the corneal endothelium of sheep using a specular microscope designed for a human eye bank. Twenty-four healthy corneas from 12 one-year-old male sheep (Ovis aries) were examined. Four central endothelial images per cornea were analyzed using a semi-automated method. Endothelial parameters were automatically calculated by the proprietary software integrated into the specular microscope, including endothelial cell density (ECD), average cell area (AVE), coefficient of variation (CV) in cell size, and percentage of hexagonal cells (HEX). Data normality was assessed using the Kolmogorov-Smirnov test, and paired comparisons between right and left eyes were performed using Student's t-test. High-quality endothelial images were successfully obtained from all corneas. The mean ECD was 3198.3 ± 428.3 cells/mm2, AVE was 318.7 ± 42.3 μm2, CV in cell size was 38.0% ± 7.9%, and HEX was 57.7%. No statistically significant differences were observed between right and left eyes for any parameter (p > 0.05). A human eye bank specular microscope allows reliable visualization and quantitative assessment of the ovine corneal endothelium, demonstrating its applicability in experimental and translational veterinary ophthalmology.

Spatially-resolved CD8+ and CD34+ computational immunohistochemistry indicators improve post-nephrectomy risk stratification in clear cell renal cell carcinoma.

The baculovirus expression vector system (BEVS) is a scalable platform used to produce recombinant adeno-associated virus vectors (rAAV) in insect cells. A major challenge in this system is reducing the formation of empty rAAV capsids, which do not contain vector DNA and lack therapeutic value. The proportion of empty capsids is influenced by the balance between the two baculovirus constructs that coinfect the producer cells: Bac-Rep-Cap, which supplies AAV replication and capsid proteins, and Bac-GOI-ITR, which delivers the therapeutic gene of interest. Digital holographic microscopy (DHM) is a label-free imaging technique that allows real-time monitoring of cell morphology in suspension cultures. Previous studies have used DHM to track cell density and baculovirus infection; however, its ability to evaluate different coinfections has not been explored. In this study, we combined DHM with machine learning to identify morphological patterns associated with various coinfections for rAAV production. Shaker-flask experiments with different Bac-Rep-Cap: Bac-GOI-ITR ratios created a dataset of cell morphologies to train a predictive classification model. When applied to real-time bioreactor measurements, the model revealed shiftsin the classification patterns related to the initial multiplicity of infection (MOI). The integration of DHM with the classification model has the potential to produce a qualitative "process fingerprint," where deviations in morphological patterns can serve as early indicators of suboptimal coinfection. Such early warning signs enable timely batch termination, reducing downstream processing of inconsistent material. Overall, DHM combined with machine learning offers a non-invasive, real-time tool for process benchmarking and quality assurance in rAAV manufacturing.

Bis(2-ethylhexyl)tetrabromophthalate disrupts thyroid hormone signaling and causes visual impairment in zebrafish larvae.

To assess the impact of omega-3 fatty acid (O3FA) supplements on tear inflammatory cytokines in dry eye patients with an omega-3 index below 4%. This randomized controlled study involved 102 dry eye patients with an omega-3 index below 4%. Participants received either four capsules of O3FAs (325 mg eicosapentaenoic acid, 175 mg docosahexaenoic acid) or a placebo containing olive oil twice daily for 6 months. Patients were evaluated at baseline and 1, 3, and 6 months. The primary outcome measured changes in tear cytokines (IL-1β, IL2, IL4, IL5, IL6, IL8, IL10, IF-γ, and TNF-α). Secondary outcomes included improvements in dry eye symptoms, Nelson grade, goblet cell density, Schirmer test values, and tear film breakup time. Group means (pretreatment, 1, 3, and 6 months) were compared using repeated measure analysis of variance. At baseline, impression cytology revealed that mRNA levels of IL-1β, IL-6, IL-8, and TNF-α were elevated by 1.6- to 2.4-fold in the O3FA group and 1.74 to 2.6-fold in the placebo group ( P = 0.123). The O3FA group experienced a statistically significant reduction ( P < 0.05) in tear cytokines. This group showed a 60% increase in the omega-3 index at 6 months, indicating high adherence to treatment. The dry eye symptom score, goblet cell density, and Nelson grade improved significantly in the O3FA group. However, these changes were not significant in the placebo group. This study underscores the potential advantages of O3FA supplementation in decreasing tear inflammatory cytokines in dry eye patients with an omega-3 index below 4%.

Changes in morphology and viability of corneal endothelium after long-term preservation in Eusol-C versus organ culture medium.

T-cell responses influence recurrence and survival in colorectal cancer. T-cell subset distributions vary by molecular phenotype and anatomic location, shaping cytotoxic or immune-cold tumor immune microenvironments. However, the T-cell contexture and their spatial proximities within preinvasive lesions are not well characterized. We analyzed sessile serrated lesions (SSL), tubulovillous/villous adenomas (TV), and tubular adenomas (TA) from 3 studies (N = 120). Whole-slide multiplex immunofluorescence was used to quantify 8 T-cell subsets [CD4+, CD8+, helper T (Th) 1 (CD4+TBX21+), Th17 (CD4+RORC+), regulatory T cells (Treg; CD4+FOXP3+), Tc1 (CD8+TBX21+), Tc17 (CD8+RORC+), and TcTreg (CD8+FOXP3+)]. Densities were compared by histology using a generalized linear mixed model with a negative binomial distribution, including an offset for total cell area and adjusting for age, sex, anatomic location, and lesion size. Nearest neighbor (NN) analyses assessed spatial proximities of T-cell pairs across lesion types. TAs and SSLs had higher CD4+ and CD8+ T-cell densities than TVs (q <0.05). Compared with TVs, TAs also had higher Th17 cell densities, whereas SSLs showed a trend toward lower Treg densities (q = 0.06). NN analysis showed greater Treg clustering in TVs than in SSLs and TAs. In contrast, TA versus SSL comparisons demonstrated predominant CD4+ clustering with Th17, Th1, and CD8+ subsets. TVs exhibited lower T-cell densities and greater Treg clustering, consistent with an immune-cold environment. SSLs and TAs were more immune-infiltrated than TVs, but TAs had higher inflammation and CD4+-dominant clustering, suggesting stronger helper coordination. Preinvasive lesions demonstrate immune and spatial heterogeneity which may have implications for primary or secondary prevention.

Extrusion‐based bioprinting has become increasingly common due to its high cell viability, compatibility with diverse crosslinking mechanisms, and ability to print at high cell densities with minimal cellular damage. However, most fabrication workflows still rely on trial-and-error optimization, leading to time-consuming, increased material costs, and reduced reproducibility. Here, we introduce a simple and yet robust mathematical model that predicts the extrusion pressure required for printing directly from rheological data, enabling the construction of a pressure-based printability window that reduces optimization time and resource use. The predicted pressures were further used to estimate wall shear stress, providing a pre-print assessment of safe-to-print conditions. The model achieved a 5–15% relative error compared with experimentally adjusted printing pressures across multiple nozzle types. All bioprinted constructs maintained >80% cell viability, and the predicted shear stress remained below reported thresholds for fibroblast safety, confirming the model’s reliability for guiding extrusion bioprinting.