Establishment of Human-Induced Pluripotent Stem Cell-Derived Neurons—A Promising In Vitro Model for a Molecular Study of Rabies Virus and Host Interaction

Rabies is a deadly viral disease caused by the rabies virus (RABV), transmitted through a bite of an infected host, resulting in irreversible neurological symptoms and a 100% fatality rate in humans. Despite many aspects describing rabies neuropathogenesis, numerous hypotheses remain unanswered and concealed. Observations obtained from infected primary neurons or mouse brain samples are more relevant to human clinical rabies than permissive cell lines; however, limitations regarding the ethical issue and sample accessibility become a hurdle for discovering new insights into virus–host interplays. To better understand RABV pathogenesis in humans, we generated human-induced pluripotent stem cell (hiPSC)-derived neurons to offer the opportunity for an inimitable study of RABV infection at a molecular level in a pathologically relevant cell type. This study describes the characteristics and detailed proteomic changes of hiPSC-derived neurons in response to RABV infection using LC-MS/MS quantitative analysis. Gene ontology (GO) enrichment of differentially expressed proteins (DEPs) reveals temporal changes of proteins related to metabolic process, immune response, neurotransmitter transport/synaptic vesicle cycle, cytoskeleton organization, and cell stress response, demonstrating fundamental underlying mechanisms of neuropathogenesis in a time-course dependence. Lastly, we highlighted plausible functions of heat shock cognate protein 70 (HSC70 or HSPA8) that might play a pivotal role in regulating RABV replication and pathogenesis. Our findings acquired from this hiPSC-derived neuron platform help to define novel cellular mechanisms during RABV infection, which could be applicable to further studies to widen views of RABV-host interaction.     

The Effects of Predrying Treatments and Different Drying Methods on Phytochemical Compound Retention and Drying Characteristics of Moringa Leaves (Moringa oleifera Lam.)

The most appropriate maturity stage of Moringa oleifera leaves was selected for drying based on phytochemical content, including quercetin and kaempferol. Desorption isotherms were developed and were best fit by the modified Henderson model. Prior to drying, samples were left untreated, blanched in boiling water, and blanched in NaHCO₃/MgO. The leaves were dried by hot air tray drying (TD) and heat pump–dehumidified drying air (HPD) at air temperatures of 40, 50, and 60°C. Alternatively, leaves were subject to microwave drying (MWD) at 150, 450, and 900 W and to freeze drying (FD). The moisture versus time data were fitted to five drying models. In general, a three-parameter model gave the best fit. The drying constant was related to the drying temperature or microwave power using an Arrhenius model. Effective moisture diffusivity (D _eff) increased with higher drying temperature, higher microwave power, or blanching treatments. Structural changes in the leaves after drying and upon rehydration were observed by scanning electron microscopy (SEM). Leaves blanched and dried using HPD at 50°C and fresh and dried using FD showed a partial breakdown of the tissue structure upon rehydration. HPD and blanching reduced the drying time by 8.3% and increased quercetin and kaempferol levels by 42.1 and 51.4%, respectively, compared to TD at 50°C. MWD provided the quickest drying followed by HPD and TD, respectively. HPD drying of M. oleifera after blanching resulted in relatively greater quality compared to TD and MWD.     

Changes during storage of dried Moringa oleifera leaves prepared by heat pump‐assisted dehumidified air drying

Moringa oleifera leaves contain phytochemicals that are retained during heat pump‐assisted dehumidified air drying. Changes in phytochemicals, antioxidant capacity and colour were evaluated at 15–35 °C, during storage of dried leaves in polypropylene (PP) or high barrier (PET/Al/PE) packaging for up to 6 months. The a_w of samples in PP increased from 0.373 to 0.669. Decreases in total phenolics were greatest at 35 °C in PP (48%) and least at 15 °C in PET/Al/PE (19%). There were few significant changes in DPPH inhibition after 2 months storage. There was little change in kaempferol and some increase in quercetin. During storage, samples became less green, suggesting breakdown in chlorophyll had occurred. The degradation of flavonoids followed first‐order kinetics. The half‐life for total flavonoids ranged from 2.13 to 1.47 months for samples stored in PP and from 2.59 to 1.83 months for samples stored in PET/Al/PE.