High demand in manufactured biologics drives the continued need for increased productivity. In this study elevated lactate metabolization resulted in improved metabolic efficiency and cellular ...productivity for a readily intensified high titer fed‐batch process. Scheduled base or lactate feeds during the stationary growth phase led to increased titers (+9% and +8% respectively) without impacting the overall growth performance. The higher lactate consumption induced by either feed strategy substituted for glutamate catabolism and consequently reduced ammonia build‐up. Direct correlation between increased titers and reduced ammonia levels was shown. Product quality attributes were impacted by both feeding strategies but could be matched with the control process by shortening the cell culture duration while maintaining titer constant.
A microstructured falling film reactor was applied to the dye-sensitized photochemical conversion of 1,5-dihydroxynaphthalene to juglone. This continuous-flow microreactor enables the efficient ...contacting of a gas and a liquid phase in combination with external irradiation by high-power LED arrays offering various wavelengths. Two sensitizers were used for the photochemical in situ generation of singlet oxygen as key step in the synthesis of the natural product juglone. The photochemical process was investigated according LED wavelength, LED power, oxygen partial pressure, reactor architecture, substrate concentration and flow rate, and optimized to a conversion of X = 97% with 99% selectivity. Based on the experimental results process parameters like quantum efficiency, productivity and space time yield were calculated and used for the evaluation of the photochemically catalyzed synthesis of juglone in continuous-flow mode.
A microstructured falling film reactor was applied to the dye-sensitized photochemical conversion of 1,5-dihydroxynaphthalene to juglone. This continuous-flow microreactor enables the efficient ...contacting of a gas and a liquid phase in combination with external irradiation by high-power LED arrays offering various wavelengths. Two sensitizers were used for the photochemical
in situ
generation of singlet oxygen as key step in the synthesis of the natural product juglone. The photochemical process was investigated according LED wavelength, LED power, oxygen partial pressure, reactor architecture, substrate concentration and flow rate, and optimized to a conversion of
X
= 97% with 99% selectivity. Based on the experimental results process parameters like quantum efficiency, productivity and space time yield were calculated and used for the evaluation of the photochemically catalyzed synthesis of juglone in continuous-flow mode.
A microstructured falling film reactor was applied to the dye-sensitized photochemical conversion of 1,5-dihydroxynaphthalene to juglone for reactor and process evaluation.
Summary
The olive (Olea europaea L. subsp. europaea) is one of the oldest and most socio‐economically important cultivated perennial crop in the Mediterranean region. Yet, its origins are still under ...debate and the genetic bases of the phenotypic changes associated with its domestication are unknown. We generated RNA‐sequencing data for 68 wild and cultivated olive trees to study the genetic diversity and structure both at the transcription and sequence levels. To localize putative genes or expression pathways targeted by artificial selection during domestication, we employed a two‐step approach in which we identified differentially expressed genes and screened the transcriptome for signatures of selection. Our analyses support a major domestication event in the eastern part of the Mediterranean basin followed by dispersion towards the West and subsequent admixture with western wild olives. While we found large changes in gene expression when comparing cultivated and wild olives, we found no major signature of selection on coding variants and weak signals primarily affected transcription factors. Our results indicated that the domestication of olives resulted in only moderate genomic consequences and that the domestication syndrome is mainly related to changes in gene expression, consistent with its evolutionary history and life history traits.
Significance Statement
We revisited the evolutionary history of the iconic olive tree by reconstructing a model of domestication and inferring the genomic changes associated with its cultivation. Although more challenging than studying annual crops, further studies of the genomic of the domestication process in perennial should broaden our understanding of the genomic bases and dynamics of adaptation.
Corneal wound healing involves communication between the different cell types that constitute the three cellular layers of the cornea (epithelium, stroma and endothelium), a process ensured in part ...by a category of extracellular vesicles called exosomes. In the present study, we isolated exosomes released by primary cultured human corneal epithelial cells (hCECs), corneal fibroblasts (hCFs) and corneal endothelial cells (hCEnCs) and determined whether they have wound healing characteristics of their own and to which point they modify the genetic and proteomic pattern of these cell types. Exosomes released by all three cell types significantly accelerated wound closure of scratch-wounded hCECs in vitro compared to controls (without exosomes). Profiling of activated kinases revealed that exosomes from human corneal cells caused the activation of signal transduction mediators that belong to the HSP27, STAT, β-catenin, GSK-3β and p38 pathways. Most of all, data from gene profiling analyses indicated that exosomes, irrespective of their cellular origin, alter a restricted subset of genes that are completely different between each targeted cell type (hCECs, hCFS, hCEnCs). Analysis of the genes specifically differentially regulated for a given cell-type in the microarray data using the Ingenuity Pathway Analysis (IPA) software revealed that the mean gene expression profile of hCECs cultured in the presence of exosomes would likely promote cell proliferation and migration whereas it would reduce differentiation when compared to control cells. Collectively, our findings represent a conceptual advance in understanding the mechanisms of corneal wound repair that may ultimately open new avenues for the development of novel therapeutic approaches to improve closure of corneal wounds.
Axicabtagene ciloleucel (axi-cel) and tisagenlecleucel (tisa-cel) have both demonstrated impressive clinical activity in relapsed/refractory (R/R) diffuse large B cell lymphoma (DLBCL). In this ...study, we analyzed the outcome of 809 patients with R/R DLBCL after two or more previous lines of treatment who had a commercial chimeric antigen receptor (CAR) T cells order for axi-cel or tisa-cel and were registered in the retrospective French DESCAR-T registry study ( NCT04328298 ). After 1:1 propensity score matching (n = 418), the best overall response rate/complete response rate (ORR/CRR) was 80%/60% versus 66%/42% for patients treated with axi-cel compared to tisa-cel, respectively (P < 0.001 for both ORR and CRR comparisons). After a median follow-up of 11.7 months, the 1-year progression-free survival was 46.6% for axi-cel and 33.2% for tisa-cel (hazard ratio (HR) = 0.61; 95% confidence interval (CI), 0.46-0.79; P = 0.0003). Overall survival (OS) was also significantly improved after axi-cel infusion compared to after tisa-cel infusion (1-year OS 63.5% versus 48.8%; HR = 0.63; 95% CI, 0.45-0.88; P = 0.0072). Similar findings were observed using the inverse probability of treatment weighting statistical approach. Grade 1-2 cytokine release syndrome was significantly more frequent with axi-cel than with tisa-cel, but no significant difference was observed for grade ≥3. Regarding immune effector cell-associated neurotoxicity syndrome (ICANS), both grade 1-2 and grade ≥3 ICANS were significantly more frequent with axi-cel than with tisa-cel. In conclusion, our matched comparison study supports a higher efficacy and also a higher toxicity of axi-cel compared to tisa-cel in the third or more treatment line for R/R DLBCL.
Anti-CD19 chimeric antigen receptor (CAR) T-cells represent a major advance in the treatment of relapsed/refractory aggressive B-cell lymphomas. However, a significant number of patients experience ...failure. Among 550 patients registered in the French registry DESCAR-T, 238 (43.3%) experienced progression/relapse, with a median follow-up of 7.9 months. At registration, 57.0% of patients presented an age-adjusted International Prognostic Index of 2 to 3, 18.9% had Eastern Cooperative Oncology Group performance status ≥2, 57.1% received >3 lines of treatment prior to receiving CAR T-cells, and 87.8% received bridging therapy. At infusion, 66% of patients presented progressive disease, and 38.9% had high lactate dehydrogenase (LDH). Failure after CAR T-cell treatment occurred after a median of 2.7 months (range: 0.2-21.5). Fifty-four patients (22.7%) presented very early failure (day D 0-D30); 102 (42.9%) had early failure (D31-D90), and 82 (34.5%) had late (>D90) failure. After failure, 154 patients (64%) received salvage treatment: 38.3% received lenalidomide, 7.1% bispecific antibodies, 21.4% targeted treatment, 11% radiotherapy, and 20% immunochemotherapy with various regimens. Median progression-free survival was 2.8 months, and median overall survival (OS) was 5.2 months. Median OS for patients failing during D0-D30 vs after D30 was 1.7 vs 3.0 months, respectively (P = .0001). Overall, 47.9% of patients were alive at 6 months, but only 18.9% were alive after very early failure. In multivariate analysis, predictors of OS were high LDH at infusion, time to CAR-T failure <D30, and high C-reactive protein at infusion. This multicentric analysis confirms the poor outcome of patients relapsing after CAR T-cell treatment, highlighting the need for further strategies dedicated to this population.
This paper focuses on the use of an electro-optic (EO) probe for the spatially and temporally resolved characterization of the electric field (E-field) associated with guided ionization waves—or ...guided streamers—propagating at atmospheric pressure. An AC high-voltage power supply (6 kV at 18 kHz) is used to generate guided ionization waves in helium with a cold atmospheric pressure plasma jet device (APPJ). The capabilities of this electro-optic technique are investigated with and without the discharge. This innovative technology will use to carry out the optical polarimetric analysis of the APPJ with simultaneous real-time measurement of longitudinal and radial components of the electric field vector. Last but not least, an application-based case study of the ionization waves propagating and interacting with the EO probe—being a solid dielectric target—is also reported. The results will bring realistic information on the potential perturbation induced by the EO probe itself on the E-field characterization at the macroscopic scale.
Graphic abstract
Variants in the ring finger protein 213 (
) gene are known to be associated with increased predisposition to cerebrovascular diseases development. Genomic studies have identified
as a major risk ...factor of Moyamoya disease in East Asian descendants. However, little is known about the RNF213 (ring finger protein 213) biological functions or its associated pathogenic mechanisms underlying Moyamoya disease.
To investigate RNF213 loss-of-function effect in endothelial cell, stable RNF213-deficient human cerebral endothelial cells were generated using the CRISPR-Cas9 genome editing technology.
In vitro assays, using RNF213 knockout brain endothelial cells, showed clear morphological changes and increased blood-brain barrier permeability. Downregulation and delocalization of essential interendothelial junction proteins involved in the blood-brain barrier maintenance, such as PECAM-1 (platelet endothelial cell adhesion molecule-1), was also observed. Brain endothelial RNF213-deficient cells also showed an abnormal potential to transmigration of leukocytes and secreted high amounts of proinflammatory cytokines.
Taken together, these results indicate that RNF213 could be a key regulator of cerebral endothelium integrity, whose disruption could be an early pathological mechanism leading to Moyamoya disease. This study also further reinforces the importance of blood-brain barrier integrity in the development of Moyamoya disease and other RNF213-associated diseases.
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