Organoids from colorectal peritoneal metastases as a platform for improving hyperthermic intraperitoneal chemotherapy.

BACKGROUND: Patients with peritoneal metastases from colorectal cancer have a poor prognosis. If the intraperitoneal tumour load is limited, patients may be eligible for cytoreductive surgery followed by hyperthermic intraperitoneal chemotherapy (HIPEC). This treatment has improved overall survival, but recurrence rates are high. The aim of this study was to create a preclinical platform for the development of more effective intraperitoneal chemotherapy strategies. METHODS: Using organoid technology, five tumour cultures were generated from malignant ascites and resected peritoneal metastases. These were used in an in vitro HIPEC model to assess sensitivity to mitomycin C (MMC) and oxaliplatin, the drugs used most commonly in HIPEC. The model was also used to test a rational combination treatment involving MMC and inhibitors of the checkpoint kinase ATR. RESULTS: MMC was more effective in eliminating peritoneal metastasis-derived organoids than oxaliplatin at clinically relevant concentrations. However, the drug concentrations required to eliminate 50 per cent of the tumour cells (IC50) were higher than the median clinical dose in two of five organoid lines for MMC, and all five lines for oxaliplatin, indicating a general resistance to monotherapy. ATR inhibition increased the sensitivity of all peritoneal metastasis-derived organoids to MMC, as the IC50 decreased 2·6-12·4-fold to well below concentrations commonly attained in clinical practice. Live-cell imaging and flow cytometric analysis showed that ATR inhibition did not release cells from MMC-induced cell cycle arrest, but caused increased replication stress and accelerated cell death. CONCLUSION: Peritoneal metastasis-derived organoids can be used to evaluate existing HIPEC regimens on an individual-patient level and for development of more effective treatment strategies. Surgical relevance Cytoreductive surgery followed by hyperthermic intraperitoneal chemotherapy (HIPEC) has improved prognosis of patients with peritoneal metastases from colorectal cancer, but disease recurrence is common. More effective and personalized HIPEC is urgently needed. Organoid technology is frequently used for drug screens, as patient-derived organoids can accurately predict clinical therapeutic response in vitro. A panel of organoids was established from peritoneal metastases from colorectal cancer and used to develop a model for testing HIPEC regimens in vitro. Patient-derived organoids differed in sensitivity to commonly used chemotherapeutics, in line with variable clinical outcomes following cytoreductive surgery-HIPEC. Combining MMC with an ATR inhibitor improved the efficacy of MMC. Peritoneal metastasis-derived organoids can be used as a platform to test novel (combination) strategies that increase HIPEC efficacy. In the future, organoids could be used to select patent-tailored HIPEC regimens.

ANTECEDENTES: Los pacientes con metástasis peritoneales (peritoneal metastasis, PM) de cáncer colorrectal tienen un mal pronóstico. Si la carga tumoral intraperitoneal es reducida, los pacientes pueden ser candidatos a cirugía citorreductora seguida de quimioterapia intraperitoneal hipertérmica (hyperthermic intraperitoneal chemotherapy, HIPEC). Este tratamiento ha mejorado la supervivencia global, pero las tasas de recidiva son altas. El objetivo de este estudio fue crear una plataforma preclínica para el desarrollo de las estrategias de quimioterapia intraperitoneal más efectivas. MÉTODOS: Mediante la utilización de la tecnología de organoides, se generaron cinco cultivos tumorales a partir de ascitis maligna y PM resecadas. Se utilizó un modelo de HIPEC in vitro para evaluar la sensibilidad a la mitomicina C (mitomycin C, MMC) y al oxaliplatino, los fármacos más utilizados en la HIPEC. El modelo también se usó para probar un tratamiento combinado de MMC e inhibidores de control inmunitario de la quinasa ATR. RESULTADOS: A concentraciones clínicamente relevantes, la MMC fue más efectiva que el oxaliplatino para eliminar los organoides derivados de PM. Sin embargo, las concentraciones de fármaco necesarias para eliminar el 50% de las células tumorales (IC50) fueron más elevadas que la mediana de la dosis clínica en 2/5 (MMC) o 5/5 (oxaliplatino) de las líneas de organoides, lo que indica una resistencia general a la monoterapia. La inhibición de ATR aumentó la sensibilidad a MMC de todos los organoides derivados de PM, ya que la IC50 disminuyó (2,6-12,4 veces) a concentraciones muy por debajo de las que se alcanzan comúnmente en la práctica clínica. Los análisis de viabilidad celular y de citometría de flujo (FACS) mostraron que la inhibición de ATR no liberaba células tras la detención del ciclo celular inducida por la MMC, sino que causaba un aumento en el estrés replicativo y muerte celular acelerada. CONCLUSIÓN: Se pueden usar los organoides derivados de PM para evaluar los regímenes HIPEC existentes a nivel del paciente individual y para desarrollar estrategias terapéuticas más efectivas.

Adaptive emergency scenery video communications using HEVC for responsive decision support in disaster incidents.

This study proposes a unifying framework for m-Health video communication systems that provides for the joint optimization of video quality, bitrate demands, and encoding time. The framework is video modality and infrastructure independent and facilitates adaptation to the best available encoding mode that satisfies underlying technology and application imposed constraints. The scalability of the proposed algorithm is demonstrated using different HEVC encoding configurations and realistic modelling of 802.11× wireless infrastructure for emergency scenery and response videos. Extensive experimentation shows that a jointly optimal solution in the encoding time, bitrate, and video quality space is feasible.

M-health medical video communication systems: an overview of design approaches and recent advances.

The emergence of the new, High Efficiency Video Coding (HEVC) standard, combined with wide deployment of 4G wireless networks, will provide significant support toward the adoption of mobile-health (m-health) medical video communication systems in standard clinical practice. For the first time since the emergence of m-health systems and services, medical video communication systems can be deployed that can rival the standards of in-hospital examinations. In this paper, we provide a thorough overview of today's advancements in the field, discuss existing approaches, and highlight the future trends and objectives.

High efficiency video coding for ultrasound video communication in m-health systems.

Emerging high efficiency video compression methods and wider availability of wireless network infrastructure will significantly advance existing m-health applications. For medical video communications, the emerging video compression and network standards support low-delay and high-resolution video transmission, at the clinically acquired resolution and frame rates. Such advances are expected to further promote the adoption of m-health systems for remote diagnosis and emergency incidents in daily clinical practice. This paper compares the performance of the emerging high efficiency video coding (HEVC) standard to the current state-of-the-art H.264/AVC standard. The experimental evaluation, based on five atherosclerotic plaque ultrasound videos encoded at QCIF, CIF, and 4CIF resolutions demonstrates that 50% reductions in bitrate requirements is possible for equivalent clinical quality.

A fast recursive shortest spanning tree for image segmentation and edge detection.

This correspondence presents a fast recursive shortest spanning tree algorithm for image segmentation and edge detection. The conventional algorithm requires a complexity of o(n(2)) for an image of n pixels, while the complexity of our approach is bounded by O(n), which is a new lower bound for algorithms of this kind. The total memory requirement of our fast algorithm is 20% smaller.

Periodic operation of immobilized live cell bioreactor for the production of candicidin.

A lumped model for cell growth and secondary metabolite production in an immobilized live cell bioreactor has been developed. This model is applied here to simulate the performance of an immobilized bioreactor under steady-state conditions and under conditions of periodically varying concentration of a growth-limiting substrate. The results of the simulation study were experimentally verified in the case of the production of the antibiotic candicidin by Streptomyces griseus in an immobilized bioreactor with forced periodic operation. The results of the studies suggest that periodically operated immobilized live cell bioreactors can provide a potent alternative for the production of non-growth-associated biochemicals, as compared to free cell fermentations, pulsed fermentations with process cycle regeneration, and nonregenerated bioreactors. This work has demonstrated that by frequent pulsing of the growth limiting nutrient, stable extended production can be obtained at high specific cellular productivities.

Modeling, optimization, and computer control of the cephalosporin C fermentation process.

In this study, a cephalosporin C producing strain, Cephalosporium acremonium (ATCC 36225), was chosen to determine the optimal conditions that maximize antibiotic production in a mixed substrate of glucose and sucrose. A model for cell growth and cephalosporin C production at different pH and temperature was developed and the associated parameters were evaluated experimentally. Pontryagin's maximum principle, in conjunction with the model, was used to predict the optimal temperature and pH control profiles to maximize the production of antibiotic.

Immobilized cell cross-flow reactor.

A cross-current flow reactor was operated using sodium alginate gel entrapped yeast cells under growth conditions. Micron-sized silica, incorporated into the biocatalyst particles (1 mm mean diameter) improved mechanical strength and internal surface adhesion. The process showed decreased productivity and stability at 35 degrees C compared to the normal study done at 30 degrees C. The increased number of cross flows diminish the product inhibition effect. The residence time distribution shows that the cross-flow bioreactor system can be approximated to either a train of backmixed fermentors in series or a plug flow fermentor with moderate axial dispersion.

Mathematical modeling of diffusion and reaction in the hydrolysis of vegetable protein in an immobilized enzyme recycle reactor.

Experimental investigation is by far the most effective approach for studying the behavior of physical systems. However, an enzymatic solubilization of vegetable protein is a complex combination of intrinsic problems, of which many are not easily adaptable to experimental investigation. Experimental designs to study enzyme vegetable protein reactions yield data which describe the extramembraneous activity of the immobilized enzyme. In a continuous recycle immobilized enzyme reactor, the microenvironment concentration of the substrate or product in the membrane phase, or the concentrations along the reactor axial length in the bulk phase are not discernible to the experimenter. However, the knowledge of such concentration profiles is important in weighing the significance of such factors as intermembrane diffusion, enzyme loading, wet membrane size, and the mode of operation of the reactor. The simulation of mathematical models, which describe the physical system within the constraints imposed, yields information which is vital to the understanding of the process occurring in the reactor. The kinetics and diffusion of an immobilized thermophilic Penicillium duponti enzyme at pH 3.4-3.7 and 50 degrees C was modeled mathematically. The kinetic parameters were evaluated by fitting a model to experimental data using nonlinear regression analysis. Simulation profiles of the effects of reactor geometry, substrate concentration, membrane thickness, and enzyme leading on the hydrolysis rate are presented. From the profiles generated by the mathematical model, the best operational reactor strategy is recommended.

Optimization of conditions and cell feeding procedures for alcohol fermentation.

Alcohol fermentation was studied with an emphasis on the separation of cell growth and alcohol production stages. Experiments were conducted to establish the optimal conditions for alcohol production in batch fermentations and to simulate continuous fermentations with cell feeding at various stages. It was found that the glucose concentration should be kept under 10% (w/v), and the temperature should be between 40 and 42.5 degrees C for maximum specific alcohol productivity. If the cell concentration is increased, a decrease in specific alcohol productivity is observed. Higher cell concentrations are needed for higher final alcohol concentrations. Among the cell feeding procedures into alcohol production stages, a cocurrent design was found to be better than recycle and countercurrent designs.

Modeling of cell viability and specific alcohol productivity.

A model has been proposed to correlate two viability tests and specific alcohol productivity of a yeast culture. Two viable populations with different activity are assumed to coexist. The difference between two populations is the ability to reproduce. Satisfactory agreement between the model and the experimental data has been observed.

Modeling the role of cyclic AMP in catabolite repression of inducible enzyme biosynthesis in microbial cells.

Modeling the role of cyclic AMP (cAMP) in catabolite repression of inducible enzyme production in microbial cells was studied. A catabolite repression index, F, was defined based on the postulation that complex formation occurs between RNA polymerase (RNAP) and DNA, and shifting from the inert form to the open form of this complex (the latter form is required for transcription) is accelerated by the cAMP.CRP complex. The catabolite repression index, F, was incorporated into model equations of mRNA production. Empirical relationships between intracellular cAMP level and medium glucose concentration were established based on experimental data and introduced into the model. Computer simulation results were obtained for a number of interesting cases. The practical utility of the proposed model was demonstrated by comparing it with the experimental results on glucose isomerase biosynthesis.