Despite advances in surgery, chemotherapy and radiotherapy, the outcomes of patients with GBM have not significantly improved. Tumor recurrence in the resection margins occurs in more than 80 % of cases indicating aggressive treatment modalities, such as gene therapy are warranted. We have examined photochemical internalization (PCI) as a method for the non-viral transfection of the cytosine deaminase (CD) suicide gene into glioma cells. The CD gene encodes an enzyme that can convert the nontoxic antifungal agent, 5-fluorocytosine, into the chemotherapeutic drug, 5-fluorouracil. Multicell tumor spheroids derived from established rat and human glioma cell lines were used as in vitro tumor models. Plasmids containing either the CD gene alone or together with the uracil phosphoribosyl transferase (UPRT) gene combined with the gene carrier protamine sulfate were employed in all experiments. PCI was performed with the photosensitizer AlPcS2a and 670 nm laser irradiance. Protamine sulfate/CD DNA polyplexes proved nontoxic but inefficient transfection agents due to endosomal entrapment. In contrast, PCI mediated CD gene transfection resulted in a significant inhibition of spheroid growth in the presence of, but not in the absence of, 5-FC. Repetitive PCI induced transfection was more efficient at low CD plasmid concentration than single treatment. The results clearly indicate that AlPcS2a-mediated PCI can be used to enhance transfection of a tumor suicide gene such as CD, in malignant glioma cells and cells transfected with both the CD and UPRT genes had a pronounced bystander effect.

The overall objective of the research was to investigate the utility of photochemical internalization (PCI) for the enhanced nonviral transfection of genes into glioma cells. The PCI-mediated introduction of the tumor suppressor gene phosphatase and tensin homolog (PTEN) or the cytosine deaminase (CD) pro-drug activating gene into U87 or U251 glioma cell monolayers and multicell tumor spheroids were evaluated. In the study reported here, polyamine-DNA gene polyplexes were encapsulated in a nanoparticle (NP) with an acid degradable polyketal outer shell. These NP synthetically mimic the roles of viral capsid and envelope, which transport and release the gene, respectively. The effects of PCI-mediated suppressor and suicide genes transfection efficiency employing either “naked” polyplex cores alone or as NP-shelled cores were compared. PCI was performed with the photosensitizer AlPcS 2a and λ=670-nm laser irradiance. The results clearly demonstrated that the PCI can enhance the delivery of both the PTEN or CD genes in human glioma cell monolayers and multicell tumor spheroids. The transfection efficiency, as measured by cell survival and inhibition of spheroid growth, was found to be significantly greater at suboptimal light and DNA levels for shelled NPs compared with polyamine-DNA polyplexes alone.

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Based on the recent success of our strained-layer superlattice (SLS)-based infrared (IR) camera that performed Earth imaging from the International Space Station (ISS) in 2019 we have built, what we consider, to be the next generation multi-band SLS imaging system. The Compact Thermal Imager (CTI) was installed on the Robotic Refueling Mission 3 (RRM3) and attached to the exterior of the ISS. From this location we were able to capture 15 million images of a multitude of fires around the globe in 2019. This unexpected trove of data initiated quite a bit of scientific interest to further utilize this imaging capability but would include features to more precisely monitor terrestrial fires and other surface phenomena. To this end, we developed a technique to install specific bandpass filters directly onto the SLS detector hybrid assembly. Utilizing this technique we have built a CTI-2 camera system with two filters, 4 and 11µm, and have made a second detector assembly with six filter bands from 4-12µm. This second system will also be used to supplement Landsat remote imaging monitoring approximate land surface temperatures, monitor evapotranspiration, sea ice and glacier dynamics. The CTI-2 camera is based on a 1,024x1,024 (1kx1k) format SLS detector hybridized to a FLIR ISC0404 readout integrated circuit (ROIC). The six band SLS focal plane array is based on the 640x512 FLIR ISC 9803 ROIC. This camera system is based on the Landsat 8 and 9 Thermal IR Sensors (TIRS) instrument and one of its purposes is to perform ground truthing for the Landsat 8/9 data at higher spectral resolution. Both Landsat TIRS instruments are dual band thermal IR sensors centered on 11 and 12µm (each with about a 1µm bandpass). Both of our SLS systems utilize a Ricor K548 cryocooler. To streamline costs and development time we used commercial optics and both commercial and custom NASA electronic components. A primary feature of these camera systems is the incorporation of specific filters to collect fire data at ~3.9µm and thermal data at ~11µm. The CTI-2 instrument is designed for 37 m /pixel spatial resolution from 410km orbit (ISS orbit). In this paper, we will present the design and performance of the focal plane, optics, electronics and mechanical structure of the dual-band CTI-2 and the focal plane performance of the six-band focal plane.