KRAS, BRAF genotyping reveals genetic heterogeneity of ovarian borderline tumors and associated implants

Background: Patients diagnosed for a serous ovarian borderline tumor (s-BOT) typically present with an excellent clinical outcome. However there have been controversies concerning the prognostic impact of so-called implants, an extra ovarian spread occurring alongside the s-BOT in certain cases. It remains obscure whether these implants actually resemble metastasis owning the same genetic pattern as the ovarian primary or whether they develop independently. Methods: The current study, in the aim of further clarifying the genetic origin of implants, assessed BRAF/KRAS hot spot mutations and the p53/p16(INK4a) immunophenotype of s-BOTs and corresponding implants (n = 49) of 15 patients by pyro-sequencing and immunostaining, respectively. Results: A significant proportion of both s-BOTs and implants showed KRAS or BRAF mutation and though p16(INK4a) was found to be abundantly expressed, p53 immunoreactivity was rather low. When genotypes of BRAF/KRAS mutated s-BOTs and corresponding implants were compared no patient presented with a fully matching mutation profile of s-BOTs and all corresponding implants. Conclusions: The current study reveals genetic heterogeneity of s-BOTs and implants, as none of the markers examined showed constant reciprocity. Hence, our findings may assist to explain the different clinical presentation of s-BOTs and implants and might encourage to applying more individualized follow up protocols

Tissue Phenomics for prognostic biomarker discovery in low- and intermediate-risk prostate cancer

Tissue Phenomics is the discipline of mining tissue images to identify patterns that are related to clinical outcome providing potential prognostic and predictive value. This involves the discovery process from assay development, image analysis, and data mining to the final interpretation and validation of the findings. Importantly, this process is not linear but allows backward steps and optimization loops over multiple sub-processes. We provide a detailed description of the Tissue Phenomics methodology while exemplifying each step on the application of prostate cancer recurrence prediction. In particular, we automatically identified tissue-based biomarkers having significant prognostic value for low-and intermediate-risk prostate cancer patients (Gleason scores 6-7b) after radical prostatectomy. We found that promising phenes were related to CD8(+) and CD68(+) cells in the microenvironment of cancerous glands in combination with the local micro-vascularization. Recurrence prediction based on the selected phenes yielded accuracies up to 83% thereby clearly outperforming prediction based on the Gleason score. Moreover, we compared different machine learning algorithms to combine the most relevant phenes resulting in increased accuracies of 88% for tumor progression prediction. These findings will be of potential use for future prognostic tests for prostate cancer patients and provide a proof-of-principle of the Tissue Phenomics approach

KRAS, BRAF genotyping reveals genetic heterogeneity of ovarian borderline tumors and associated implants

Background: Patients diagnosed for a serous ovarian borderline tumor (s-BOT) typically present with an excellent clinical outcome. However there have been controversies concerning the prognostic impact of so-called implants, an extra ovarian spread occurring alongside the s-BOT in certain cases. It remains obscure whether these implants actually resemble metastasis owning the same genetic pattern as the ovarian primary or whether they develop independently. Methods: The current study, in the aim of further clarifying the genetic origin of implants, assessed BRAF/KRAS hot spot mutations and the p53/p16(INK4a) immunophenotype of s-BOTs and corresponding implants (n = 49) of 15 patients by pyro-sequencing and immunostaining, respectively. Results: A significant proportion of both s-BOTs and implants showed KRAS or BRAF mutation and though p16(INK4a) was found to be abundantly expressed, p53 immunoreactivity was rather low. When genotypes of BRAF/KRAS mutated s-BOTs and corresponding implants were compared no patient presented with a fully matching mutation profile of s-BOTs and all corresponding implants. Conclusions: The current study reveals genetic heterogeneity of s-BOTs and implants, as none of the markers examined showed constant reciprocity. Hence, our findings may assist to explain the different clinical presentation of s-BOTs and implants and might encourage to applying more individualized follow up protocols

An asymmetric organocatalytic aldol reaction of a hydrophobic aldehyde in aqueous medium running in flow mode

\u3cp\u3eReaction conditions have been identified to conduct a one-pot asymmetric organocatalytic aldol reaction with a hydrophobic substrate in aqueous medium via a process running in flow mode. By employing a mixture of water and 2-propanol, a hydrophobic aldehyde and 3.6 mol% of an organocatalyst, this microreactor process affords the desired aldol adduct with a conversion of 74% and an enantioselectivity of 89% after a reaction time of 60 minutes.\u3c/p\u3

An Asymmetric Organocatalytic Aldol Reaction of a Hydrophobic Aldehyde in Aqueous Medium Running in Flow Mode

Reaction conditions have been identified to conduct a one-pot asymmetric organocatalytic aldol reaction with a hydrophobic substrate in aqueous medium via a process running in flow mode. By employing a mixture of water and 2-propanol, a hydrophobic aldehyde and 3.6 mol% of an organocatalyst, this microreactor process affords the desired aldol adduct with a conversion of 74% and an enantioselectivity of 89% after a reaction time of 60 minutes

Ionic Liquid/Water Continuous-Flow System with Compartmentalized Spaces for Automatic Product Purification of Biotransformation with Enzyme Recycling

Deng Q, Nam Nghiep Tran NNT, Asrami MR, Schober L, Gröger H, Hessel V. Ionic Liquid/Water Continuous-Flow System with Compartmentalized Spaces for Automatic Product Purification of Biotransformation with Enzyme Recycling. INDUSTRIAL & ENGINEERING CHEMISTRY RESEARCH. 2020;59(48):21001-21011.In this work, we developed a biphasic designer solvent system for enzymatic biotransformation, to demonstrate automatic purification and enzyme reuse, in the frame of a new process concept reported recently (solvent-enabled factory, One-Flow project). "Automatic" refers to instantaneous operation by preferential solubility between two immiscible phases, which does not require (sophisticated) process control ("automated"). The reaction studied is lipase-catalyzed hydrolysis of 4-nitrophenyl acetate. As a designer solvent, the class of ionic liquids (ILs) has been chosen, and their usage is largely documented in the biocatalysis literature. Three ILs are chosen; all with the 1-butyl-3-methyl-imidazolium cation and differing in their anions, being tetrafluoroborate, bis(trifluoromethylsulfonyl)-imide, and hexafluoro-phosphate. The first IL forms a monophasic system and thus was left out of consideration. The other two form the desired biphasic system, when being contacted with water, respectively. The operation of that system is hampered by foaming, that is, the formation of an interfacial emulsion layer as the third phase, which makes phase separation difficult. Therefore, we investigated in detail the phase behavior of the batch and flow-processed fluid systems under various process conditions. Batch processing, which causes tremendous foaming, needs intense stirring because of the high IL viscosity. Continuous-flow reactors provide an alternative because they stir more softly by their shear-enforced convection in their liquid slugs. As a result, they do not show foaming, and therefor; separation in phases is facile. With that issue solved, we report here for the continuous-flow biocatalytic reaction that we achieved high-level product purification and (three times) recycling of the enzyme

Screening of functional solvent system for automatic aldehyde and ketone separation in aldol reaction: A combined COSMO-RS and experimental approach

Zhang C, Song Z, Jin C, et al. Screening of functional solvent system for automatic aldehyde and ketone separation in aldol reaction: A combined COSMO-RS and experimental approach. CHEMICAL ENGINEERING JOURNAL. 2020;385: 123399.An integrated reaction-separation process based on only solvents is presented, and thus without the need for related technical equipment (reactor-separator). Both product purification and reactant recovery are achieved automatically for an asymmetric aldol reaction employing a biphasic solvent system as compartmentalizing soft matter. Firstly, COSMO-RS based simulation is introduced as a theoretical guidance in the solvent selection for the reactant 3-chlorobenzaldehyde and non-solvent selection for the R-aldol product, (R) 4 (3 chlorophenyl)-4-hydroxybutan-2-one. This encompasses solubility as core critical parameter, as well as chemo-physical properties and environmental profiling. Such criteria-cascaded screening could effectively reduce a 7665 solvents-database into 1 candidate solvent, which is dodecane, before experimental process assessment. Secondly, this screening's top candidate was validated as the best reaction solvent by first a solubility test and then by a batch reaction, in which a conversion of 69% was achieved. As desired, the mono-phase reaction yielded spontaneously the product layer and the separate dodecane phase as the second layer, which indeed allowed facile separation of the product from the residual reactant. In a third step, a segmented flow process was developed giving a highest product yield of 63% and a total conversion 92% respectively after a 2 h residence time