Phoenix: A Portable, Battery-Powered, and Environmentally Controlled Platform for Long-Distance Transportation of Live-Cell Cultures.

Despite the advent of advanced therapy medicinal products (ATMPs) in regenerative medicine, gene therapy, cell therapies, tissue engineering, and immunotherapy, the availability of treatment is limited to patients close to state-of-the-art facilities. The SCORPIO-V Division of HNu Photonics has developed the Phoenix-Live Cell Transport, a battery-operated mobile incubator designed to facilitate long-distance transportation of living cell cultures from GMP facilities to remote areas for increased patient accessibility to ATMPs. This work demonstrates that Phoenix (patent pending) is a superior mechanism for transporting living cells compared to the standard method of shipping frozen cells on dry ice (-80°C) or in liquid nitrogen (-150°C), which are destructive to the biology as well as a time consuming process.

Azelnidipine Attenuates the Oxidative and NFκB Pathways in Amyloid-β-Stimulated Cerebral Endothelial Cells.

Cerebral amyloid angiopathy (CAA), a condition depicting cerebrovascular accumulation of amyloid β-peptide (Aβ), is a common pathological manifestation in Alzheimer's disease (AD). In this study, we investigated the effects of Azelnidipine (ALP), a dihydropyridine calcium channel blocker known for its treatment of hypertension, on oligomeric Aβ (oAβ)-induced calcium influx and its downstream pathway in immortalized mouse cerebral endothelial cells (bEND3). We found that ALP attenuated oAβ-induced calcium influx, superoxide anion production, and phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) and calcium-dependent cytosolic phospholipase A (cPLA).

Cytosolic Phospholipase A Facilitates Oligomeric Amyloid-β Peptide Association with Microglia via Regulation of Membrane-Cytoskeleton Connectivity.

Cytosolic phospholipase A (cPLA) mediates oligomeric amyloid-β peptide (oAβ)-induced oxidative and inflammatory responses in glial cells. Increased activity of cPLA has been implicated in the neuropathology of Alzheimer's disease (AD), suggesting that cPLA regulation of oAβ-induced microglial activation may play a role in the AD pathology. We demonstrate that LPS, IFNγ, and oAβ increased phosphorylated cPLA (p-cPLA) in immortalized mouse microglia (BV2).

The role of protein hydrophobicity in conformation change and self-assembly into large amyloid fibers.

It has been found that a short hydrophobic "template" peptide and a larger α-helical "adder" protein cooperatively self-assemble into micrometer sized amyloid fibers. Here, a common template of trypsin hydrolyzed gliadin is combined with six adder proteins (α-casein, α-lactalbumin, amylase, hemoglobin, insulin, and myoglobin) to determine what properties of the adder protein drive amyloid self-assembly. Utilizing Fourier Transform-Infrared (FT-IR) spectroscopy, the Amide I absorbance reveals that the observed decrease in α-helix with time is approximately equal to the increase in high strand density β-sheet, which is indicative of amyloid formation.

Characterization of large amyloid fibers and tapes with Fourier transform infrared (FT-IR) and Raman spectroscopy.

Amyloids are self-assembled protein structures implicated in a host of neurodegenerative diseases. Organisms can also produce "functional amyloids" to perpetuate life, and these materials serve as models for robust biomaterials. Amyloids are typically studied using fluorescent dyes, Fourier transform infrared (FT-IR), or Raman spectroscopy analysis of the protein amide I region, and X-ray diffraction (XRD) because the self-assembled β-sheet secondary structure of the amyloid can be easily identified with these techniques.

Evolution of the amyloid fiber over multiple length scales.

The amyloid is a natural self-assembled peptide material comparable in specific stiffness to spider silk and steel. Throughout the literature there are many studies of the nanometer-sized amyloid fibril; however, peptide mixtures are capable of self-assembling beyond the nanometer scale into micrometer-sized fibers. Here, atomic force microscopy (AFM) and scanning electron microscopy (SEM) are used to observe the self-assembly of the peptide mixtures in solution for 20 days and the fibers upon drying.

Peptide mixtures can self-assemble into large amyloid fibers of varying size and morphology.

Peptide mixtures spontaneously formed micrometer-sized fibers and ribbons from aqueous solution. Hydrolyzed gliadin produced short, slightly elliptical fibers while hydrolyzed wheat gluten, a mixture of gliadin and glutenin, formed round fibers of similar size. Mixing hydrolyzed gliadin with increasing molar amounts of myoglobin or amylase resulted in longer, wider fibers that transitioned from round to rectangular cross section.