Towards Automation in 3D Cell Culture: Selective and Gentle High-Throughput Handling of Spheroids and Organoids via Novel Pick-Flow-Drop Principle.

3D cell culture is becoming increasingly important for mimicking physiological tissue structures in areas such as drug discovery and personalized medicine. To enable reproducibility on a large scale, automation technologies for standardized handling are still a challenge. Here, a novel method for fully automated size classification and handling of cell aggregates like spheroids and organoids is presented. Using microfluidic flow generated by a piezoelectric droplet generator, aggregates are aspirated from a reservoir on one side of a thin capillary and deposited on the other side, encapsulated in free-flying nanoliter droplets to a target. The platform has aggregate aspiration and plating efficiencies of 98.1% and 98.4%, respectively, at a processing throughput of up to 21 aggregates per minute. Cytocompatibility of the method is thoroughly assessed with MCF7, LNCaP, A549 spheroids and colon organoids, revealing no adverse effects on cell aggregates as shear stress is reduced compared to manual pipetting. Further, generic size-selective handling of heterogeneous organoid samples, single-aggregate-dispensing efficiencies of up to 100% and the successful embedding of spheroids or organoids in a hydrogel with subsequent proliferation is demonstrated. This platform is a powerful tool for standardized 3D in vitro research.

Epidural Extramedullary Plasmacytoma Successfully Treated with Surgical Excision and Bortezomib, Cyclophosphamide and Dexamethasone.

Plasma cell neoplasms are common, accounting for more than 1% of all malignancies. Its most common form is multiple myeloma, but others, such as extramedullary plasmacytoma (EMP), exist. Spinal cord compression secondary to these pathologies is not uncommon, however, adjacent bone involvement is usually present. Spinal cord compression caused by isolated epidural EMP is extremely rare, with only one case reported to date. We describe the case of a 75-year-old female patient that presented with paraparesis, due to an isolated epidural EMP associated with light chain amyloidosis. She was treated with surgical decompression and neoadjuvant chemotherapy, and is currently with a 15-month disease-free period.

Holographic PIV/PTV for nano flow rates-A study in the 70 to 200 nL/min range.

Accurately measuring flow rates is a key requirement in many medical applications such as infusion and drug delivery systems. A major drawback of current systems is the low resolution of the sensors in the low flow rate regime. In this article, we present a method based on Holographic PIV/PTV that has been used for the first time to measure flow rates in the range of a few nL/min. Our method requires a very simple setup that combines lensless holography with particle velocimetry. For flow rates in the 70 to 200 nL/min range, the highest uncertainty was 5.6% (coverage factor k=2). This is an open-source project; the CAD designs and software source code are available at https://github.com/gui-miotto/holovel.

Calibration methods for flow rates down to 5 nL/min and validation methodology.

Improving the accuracy and enabling traceable measurements of volume, flow, and pressure in existing drug delivery devices and in-line sensors operating at very low flow rates is essential in several fields of activities and specially in medical applications. This can only be achieved through the development of new calibrationmethods and by expanding the existing metrological infrastructure to perform micro-flow and nano-flow measurements. In this paper, we will investigate new traceable techniques for measuring flow rate, from 5 nL/min to 1,500 nL/min and present the results of an inter-comparison between nine laboratories for the calibration of two different flow meters and a syringe pump.

Deep Learning-Assisted Nephrotoxicity Testing with Bioprinted Renal Spheroids.

We used arrays of bioprinted renal epithelial cell spheroids for toxicity testing with cisplatin. The concentration-dependent cell death rate was determined using a lactate dehydrogenase assay. Bioprinted spheroids showed enhanced sensitivity to the treatment in comparison to monolayers of the same cell type. The measured dose-response curves revealed an inhibitory concentration of the spheroids of IC 50 = 9 ± 3 µM in contrast to the monolayers with IC 50 = 17 ± 2 µM. Fluorescent labeling of a nephrotoxicity biomarker, kidney injury molecule 1 indicated an accumulation of the molecule in the central lumen of the spheroids. Finally, we tested an approach for an automatic readout of toxicity based on microscopic images with deep learning. Therefore, we created a dataset comprising images of single spheroids, with corresponding labels of the determined cell death rates for training. The algorithm was able to distinguish between three classes of no, mild, and severe treatment effects with a balanced accuracy of 78.7%.

Aggressive vertebral hemangiomas - case series and literature review / Hemangiomas vertebrais de comportamento agressivo - série de casos e revisão da literatura / Hemangiomas vertebrales de comportamiento agresivo - serie de casos y revisión de la literatura

ABSTRACT Objectives To present a series of aggressive hemangiomas of the institution, with a review of the management options described in the literature. Methods This is a retrospective survey of aggressive vertebral hemangiomas treated by the service in the last 10 years, with histological confirmation of the diagnosis and a minimum follow-up of 1 year. The case analysis and literature review were conducted with emphasis on treatment options for these injuries. Results Seven cases were found, three with pain and four with severe neurological deficits. Two patients were treated with open decompression, one with open decompression and cementation, one with open decompression and arthrodesis, one with biopsy and cementation, one with percutaneous biopsy, and one with open biopsy followed by decompression surgery. All patients underwent radiotherapy. There was a significant regression of presentation deficits, but one patient developed an irreversible deficit during treatment. There were no recurrences or late complications in the follow-up period. Conclusions Surgical decompression in patients with significant neurological deficit is a point of consensus in the literature. Subtotal resection followed by radiation therapy was effective in treating deficits and controlling pathology. Cases manifesting pain only can be managed with minimally invasive techniques, whether or not they are followed by radiotherapy. Level of evidence IV; Therapeutic study of case series.

RESUMO Objetivo Apresentar a série de hemangiomas agressivos da instituição, com uma revisão das opções de manejo descritas na literatura. Métodos Trata-se de um levantamento retrospectivo dos hemangiomas vertebrais agressivos atendidos pelo serviço nos últimos 10 anos, com confirmação histológica do diagnóstico e seguimento mínimo de um ano. Foi feita análise dos casos e revisão da literatura, com ênfase nas opções de tratamento dessas lesões. Resultados Foram encontrados sete casos, três com dor e quatro com déficits neurológicos graves. Dois pacientes foram tratados com descompressão aberta, um com descompressão aberta e cimentação, um com descompressão aberta e artrodese, um com biópsia e cimentação, um com biópsia percutânea e um com biópsia aberta seguida de cirurgia descompressiva. Todos os pacientes foram submetidos à radioterapia. Houve regressão significativa dos déficits da apresentação, mas uma paciente desenvolveu déficit irreversível durante o tratamento. Não houve recidivas ou complicações tardias no período de acompanhamento. Conclusões A descompressão cirúrgica nos pacientes com déficit neurológico significativo é um ponto de consenso na literatura. A ressecção subtotal seguida de radioterapia foi eficaz no tratamento dos déficits e no controle da patologia. Casos manifestos apenas por dor podem ser manejados com técnicas minimamente invasivas, seguidas ou não de radioterapia. Nível de evidência IV; Estudo terapêutico de série de casos.

RESUMEN Objetivo Presentar la serie de hemangiomas agresivos de la institución, con una revisión de las opciones de manejo descritas en la literatura. Métodos Se trata de un levantamiento retrospectivo de los hemangiomas vertebrales agresivos atendidos por el servicio en los últimos diez años, con confirmación histológica del diagnóstico y un seguimiento mínimo de un año. Fue hecho análisis de los casos y revisión de la literatura, con énfasis en las opciones de tratamiento de esas lesiones. Resultados Fueron encontrados siete casos, tres con dolor y cuatro con déficits neurológicos graves. Dos pacientes fueron tratados con descompresión abierta, uno con descompresión abierta y cementación, uno con descompresión abierta y artrodesis, uno con biopsia y cementación, uno con biopsia percutánea y uno con biopsia abierta seguida de cirugía descompresiva. Todos los pacientes fueron sometidos a radioterapia. Hubo regresión significativa de los déficits de presentación, pero una paciente desarrolló déficit irreversible durante el tratamiento. No hubo recidivas o complicaciones tardías en el período de acompañamiento. Conclusiones La descompresión quirúrgica en los pacientes con déficit neurológico significativo es un punto de consenso en la literatura. La resección subtotal seguida de radioterapia fue eficaz en el tratamiento de los déficits y en el control de la patología. Los casos manifestados solo por dolor pueden ser manejados con técnicas mínimamente invasivas, seguidas o no de radioterapia. Nivel de evidencia IV; Estudio terapéutico de serie de casos.

A review on techniques for tremor recording and quantification.

Tremor is the most common movement disorder and differs from other disorders by its repetitive, stereotyped movements, with regular frequency and amplitude. The three most frequent pathological forms of it are the essential tremor (ET), the Parkinson's disease (PD) tremor, and the enhanced physiological tremor. The ET and PD tremor affect the older population mostly. Although there are cases of tremor reported since ancient times, there is currently no consensus about its causes or about its main differential characteristics. In this article, we present a review of the methods more frequently used in measurement and analysis of tremor and the difficulties encountered in the research for the identification of methodologies that allow a significant advance in the study of tremor.