RESUMO
Induced pluripotent stem cell (iPSC) derived endothelial cells (iECs) have emerged as a promising tool for studying vascular biology and providing a platform for modelling various vascular diseases, including those with genetic origins. Currently, primary ECs are the main source for disease modelling in this field. However, they are difficult to edit and have a limited lifespan. To study the effects of targeted mutations on an endogenous level, we generated and characterized an iPSC derived model for venous malformations (VMs). CRISPR-Cas9 technology was used to generate a novel human iPSC line with an amino acid substitution L914F in the TIE2 receptor, known to cause VMs. This enabled us to study the differential effects of VM causative mutations in iECs in multiple in vitro models and assess their ability to form vessels in vivo. The analysis of TIE2 expression levels in TIE2L914F iECs showed a significantly lower expression of TIE2 on mRNA and protein level, which has not been observed before due to a lack of models with endogenous edited TIE2L914F and sparse patient data. Interestingly, the TIE2 pathway was still significantly upregulated and TIE2 showed high levels of phosphorylation. TIE2L914F iECs exhibited dysregulated angiogenesis markers and upregulated migration capability, while proliferation was not affected. Under shear stress TIE2L914F iECs showed reduced alignment in the flow direction and a larger cell area than TIE2WT iECs. In summary, we developed a novel TIE2L914F iPSC-derived iEC model and characterized it in multiple in vitro models. The model can be used in future work for drug screening for novel treatments for VMs.
Assuntos
Células Endoteliais , Técnicas de Introdução de Genes , Células-Tronco Pluripotentes Induzidas , Receptor TIE-2 , Humanos , Células-Tronco Pluripotentes Induzidas/metabolismo , Receptor TIE-2/genética , Receptor TIE-2/metabolismo , Células Endoteliais/metabolismo , Mutação/genética , Sistemas CRISPR-Cas/genética , Malformações Vasculares/genética , Malformações Vasculares/patologia , Malformações Vasculares/metabolismoRESUMO
The development of new therapies is hampered by the lack of predictive, and patient-relevant in vitro models. Organ-on-chip (OOC) technologies can potentially recreate physiological features and hold great promise for tissue and disease modeling. However, the non-standardized design of these chips and perfusion control systems has been a barrier to quantitative high-throughput screening (HTS). Here we present a scalable OOC microfluidic platform for applied kinetic in vitro assays (AKITA) that is applicable for high, medium, and low throughput. Its standard 96-well plate and 384-well plate layouts ensure compatibility with existing laboratory workflows and high-throughput data collection and analysis tools. The AKITA plate is optimized for the modeling of vascularized biological barriers, primarily the blood-brain barrier, skin, and lung, with precise flow control on a custom rocker. The integration of trans-epithelial electrical resistance (TEER) sensors allows rapid and repeated monitoring of barrier integrity over long time periods. Together with automated liquid handling and compound permeability testing analyses, we demonstrate the flexibility of the AKITA platform for establishing human-relevant models for preclinical drug and precision medicine's efficacy, toxicity, and permeability under near-physiological conditions.
RESUMO
Alpinia nelumboides Nob.Tanaka, T.T.K.Van & V.Hoang is the new Alpninia species discovered in Vietnam in 2023. Herein, we first hydrodistillated its pseudo-stems and rhizomes to obtain its essential oils, PS-EO and RH-EO. Their volatile compounds and total polyphenols were analysed by gas chromatography-mass spectrometry and the Folin-Ciocalteu method, respectively. Antioxidant activities were determined using four different approaches. The results showed that PS-EO and RH-EO contained 40 and 31 compounds, accounting for 99.78% and 99.45% of their compositions, respectively. The contents of polyphenols and monoterpenes in PS-EO were higher than in RH-EO. RH-EO displayed weaker scavenging activities (17.40-19.53%) than PS-EO (30.81-44.08%). PS-EO also showed higher ferric and cupric reducing powers, with EC50 values of 3.50-5.30 mg/mL smaller than RH-EO's EC50 values of 19.0-23.0 mg/mL. These results first revealed the phytochemical profile and antioxidant activities of EOs from A. nelumboides.
RESUMO
Microfluidics are expected to revolutionize the healthcare industry especially in developing countries since it would bring portable, easy-to-use, self-contained diagnostic devices to places with limited access to healthcare. To date, however, microfluidics has not yet been able to live up to these expectations. One non-negligible factor can be attributed to inaccessible prototyping methods for researchers in low-resource settings who are unable to afford expensive equipment and/or obtain critical reagents and, therefore, unable to engage and contribute to microfluidics research. In this paper, we present methods to create microfluidic devices that reduce initial costs from hundreds of thousands of dollars to about $6000 by using readily accessible consumables and inexpensive equipment. By including the scientific community most embedded and aware of the requirements of healthcare in developing countries, microfluidics will be able to increase its reach in the research community and be better informed to provide relevant solutions to global healthcare challenges.
RESUMO
60 patients with chronic B hepatitis were treated at The Hospital No 108, using At Can Ninh, a preparation of traditional medicine. Some clinical symptoms were improved rapidly such as themia anotexia, jaundive, clark yellow urine. In comparing with control subjects, after 2-3 weeks treatment, SGOT, SGPT in blood were reduced more quickly. The preparation also reduce it Bs Ag, HBV- DNA level and serum converting rate was higher 14,3% verous 0% with p<0,05 after 3 months treatment. Some uncomfortable manifestation such as pylori pain and nausea in 2/30 patients were disappeared immediately after the cease of medicine.