Nanoprobes (United States)

Abstract Current gold standard for absolute quantitation of a specific DNA sequence is droplet digital PCR (ddPCR), which has been applied to gene copy number variation (CNV) detection. However, the number of quantitation modules in ddPCR has been limited by fluorescence channels, which thus limits the CNV sensitivity due to sampling error following Poisson distribution. Here we develop a PCR-based molecular barcoding NGS approach, quantitative amplicon sequencing (QASeq), for accurate absolute quantitation which is compatible with high multiplexing to include over 200 quantitation modules. By attaching barcodes to individual target molecules with high efficiency, 2-plex QASeq exhibits higher and more consistent conversion yield than ddPCR in absolute molecule count quantitation. Multiplexed QASeq improves CNV sensitivity to allow confident distinguishment of 2.05 ploidy from normal 2.00 ploidy. We applied multiplexed QASeq to serial longitudinal plasma cfDNA samples from patients with metastatic ERBB2+ (HER2+) breast cancer seeking association with tumor progression. We further show an RNA QASeq panel for targeted expression profiling on a wide range of RNA samples including tumor formalin-fixed, paraffin-embedded (FFPE) RNA.

Abstract About 30% of breast cancers metastasize to brain; those widely disseminated are fatal typically in 3-4 months, even with the best available surgery, drugs, and radiotherapy. To address this dire situation, we have developed iodine nanoparticles (INPs) that target brain tumors after intravenous (IV) injection. The iodine then absorbs X-rays during radiotherapy (RT), creating free radicals and local tumor damage, effectively boosting the local RT dose at the tumor. Efficacy was tested using the very aggressive human triple negative breast cancer (TNBC, MDA-MB-231 cells) growing in the brains of athymic nude mice. With a well-tolerated non-toxic IV dose of the INPs (7 g iodine/kg body weight), tumors showed a heavily iodinated rim surrounding the tumor having an average uptake of 2.9% iodine by weight (peaks at 4.5%), calculated to provide dose enhancement factors of ~5.5 (peaks at 8.0) – the highest ever reported for any radio-enhancing agents. With 15-Gy, single dose RT, all animals died by 72 days; INP pretreatment resulted in longer-term remissions with 40% of mice surviving 150 days and 30% surviving > 280 days (Hainfeld et al., 2020). Fluorescence confocal microscopy revealed most INP staining co-localized with CD31 in the tumor center and tumor periphery. Greatest INP and CD31 staining was in the tumor periphery, the region of increased Micro CT contrast. Tumor cells are seen to line irregularly-shaped spaces (ISS) with INP and CD31 staining very close to or on the tumor cell surface and with PAS stain on their boundary and may represent a unique form of CD31-expressing vascular mimicry in intracerebral 231-tumors. INP and CD31 co-staining is also seen around ISS formed around tumor cells migrating on CD31 positive blood-vessels. We hypothesize that breast cancer cells secrete a CD31 containing scaffold to which IV-injected INPs bind; tumor cells proliferate along the scaffold forming the boundaries of the ISS creating a form of vascular mimicry. The significant radiation dose enhancement to the prolific INP-binding ISS found throughout the tumor but concentrated in the tumor rim, may contribute significantly to the life extensions observed after INP-RT; vascular mimicry could represent a new nanoparticle, particularly INP, tumor-homing target. Citation Format: Sharif M Ridwan, James F Hainfeld, Yaroslav Stanishevskiy, Vanessa Ross, Henry M Smilowitz. Novel iodine nanoparticle micro-localization and resultant radiotherapy-enhancement of triple negative human breast cancer growing in the brains of athymic nude mice [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PS14-16.

A huge amount of thermal energy is available close to material surfaces in radiative and non-radiative states, which can be useful for matter characterization or for energy devices. One way to harness this near-field energy is to scatter it to the far field. Another way is to bring absorbers close to thermal emitters, and the advent of a full class of novel photonic devices exploiting thermal photons in the near field has been predicted in the last two decades. However, efficient heat-to-electricity conversion of near-field thermal photons, i.e. the seminal building block, could not be achieved experimentally until now. Here, by approaching a micron-sized infrared photovoltaic cell at nanometric distances from a hot surface, we demonstrate conversion efficiency up to 14% leading to unprecedented electrical power density output (7500 W.m-2), orders of magnitude larger than all previous attempts. This proof of principle is achieved by using hot graphite microsphere emitters (~800 K) and indium antimonide cells, whose low bandgap energy matches the emitter infrared spectrum and which are specially designed for the near field. These results pave the way for efficient photoelectric detectors converting thermal photons directly in the near field. They also highlight that near-field thermophotovoltaic converters, which harvest radiative thermal energy in a contactless manner, are now competing with other energy-harvesting devices, such as thermoelectrics, over a large range of heat source temperatures.

Gliomas and other brain tumors have evaded durable therapies, ultimately causing about 20% of all cancer deaths. Tumors are widespread in the brain at time of diagnosis, limiting surgery and radiotherapy effectiveness. Drugs are also poorly effective. Radiotherapy (RT) is limited by dose to normal tissue. However, high-atomic-number elements absorb X-rays and deposit the absorbed dose locally, even doubling (or more) the local dose. Previously we showed that gold nanoparticles (AuNPs) with RT could eradicate some brain tumors in mice and many other preclinical studies confirmed AuNPs as outstanding radioenhancers. However, impediments to clinical translation of AuNPs have been poor clearance, skin discoloration, and cost. We therefore developed iodine nanoparticles (INPs) that are almost colorless, non-toxic, lower cost, and have reasonable clearance, thus overcoming major drawbacks of AuNPs. Here we report the use of iodine nanoparticle radiotherapy (INRT) in treating advanced human gliomas (U87) grown orthotopically in nude mice resulting in a more than a doubling of median life extension compared to RT alone. Significantly, INRT also enhanced the efficacy of chemotherapy when it was combined with the chemotherapeutic agent Doxil, resulting in some longer-term survivors. While ongoing optimization studies should further improve INRT, clinical translation appears promising.

purified protein fraction). The low photo-labeling can be explained based on inherent low PAL efficiencies

The reduction of fluid drag is an important issue in many fluid flow applications at the micro/nanoscale. Boundary slip is believed to affect fluid drag. Slip length has been measured on various surfaces with different degrees of hydrophobicity and oleophobicity immersed in various liquids of scientific interest. Surface charge has been found to affect slip length in water and electrolytes. However, there are no studies on the effect of surface charge on slip at solid-oil interfaces. This study focuses on the effect of surface charge on the boundary slip of superoleophilic, oleophilic, oleophobic, and superoleophobic surfaces immersed in deionized (DI) water and hexadecane and ethylene glycol, based on atomic force microscopy (AFM). The surface charge was changed by applying a positive electric field to the solid-liquid interface, and by using liquids with different pH values. The results show that slip length increases with an increase in applied positive electric field voltage. Slip length also increases with a decrease in the pH of the solutions. The change in slip length is dependent on the absolute value of the surface charge, and a larger surface charge density results in a smaller slip length. In addition, the surface charge density at different solid-liquid interfaces is related to the dielectric properties of the surface. The underlying mechanisms are analyzed.

Extended abstract of a paper presented at Microscopy and Microanalysis 2010 in Portland, Oregon, USA, August 1 – August 5, 2010.

The purpose of this study is to test the hypothesis that gold nanoparticle (AuNP, nanogold)-enhanced radiation therapy (nanogold radiation therapy, NRT) is efficacious when treating the radiation resistant and highly aggressive mouse head and neck squamous cell carcinoma model, SCCVII, and to identify parameters influencing the efficacy of NRT. Subcutaneous (sc) SCCVII leg tumors in mice were irradiated with x-rays at the Brookhaven National Laboratory (BNL) National Synchrotron Light Source (NSLS) with and without prior intravenous (iv) administration of AuNPs. Variables studied included radiation dose, beam energy, temporal fractionation and hyperthermia. AuNP-mediated NRT was shown to be effective for the sc SCCVII model. AuNPs were more effective at 42 Gy than at 30 Gy (both at 68 keV median beam energy) compared to controls without gold. Similarly, at 157 keV median beam energy, 50.6 Gy NRT was more effective than 44 Gy NRT. At the same radiation dose (∼42 Gy), 68 keV was more effective than 157 keV. Hyperthermia and radiation therapy (RT) were synergistic and AuNPs enhanced this synergy, thereby further reducing TCD50 s (tumor control dose 50%) and increasing long-term survivals. It is concluded that gold nanoparticles enhance the radiation therapy of a radioresistant mouse squamous cell carcinoma. The data show that radiation dose, energy and hyperthermia influence efficacy and better define the potential utility of gold nanoparticles for cancer x-ray therapy.

Ultrasensitive bright field in situ hybridization assays using enzyme metallography (EnzMet) have been developed and validated, but little is known regarding the applicability of EnzMet for immunophenotypic detection of protein via IHC. Superior resolution via discrete metallographic deposits offers the potential for enhancing high-resolution immunophenotyping. Using high-complexity tissue microarrays (TMAs), 88 common solid tumors were evaluated by automated EnzMet (Nanoprobes and Ventana). Targets were chosen to assess the ability of EnzMet to specifically localize encoded antigens in the nucleus (estrogen receptor), cytoplasm (cytokeratins), and cytoplasmic membrane (HER2) in TMAs. Results were compared with conventional IHC diaminobenzidine (DAB) immunostaining. There was full concordance between the EnzMet and conventional IHC results. Furthermore, the EnzMet reaction products did not appreciably diffuse, were dense and sharply defined, and provided excellent high-resolution differentiation of cellular compartments in paraffin sections for the nuclear, cytoplasmic, and cell membrane-localized antigens evaluated. The higher density of elemental silver deposited during enzyme metallography permitted evaluation of core immunophenotypes at a relatively low magnification, allowing more tissue to be screened in an efficient manner. This preliminary study shows the utility of using enzyme metallography for high-resolution immunophenotyping in TMAs.