Chemotherapeutics-Loaded Poly(Dopamine) Core-Shell Nanoparticles for Breast Cancer Treatment.

Chemophotothermal therapy is an emerging treatment of metastatic and drug-resistant cancer anomalies. Among various photothermal agents tested, poly(dopamine) provides an excellent biocompatible alternative that can be used to develop novel drug delivery carriers for cancer treatment. This study explores the synthesis of starch-encapsulated, poly(dopamine)-coated core-shell nanoparticles in a one-pot synthesis approach and by surfactant-free approach. The nanoparticles produced are embellished with polymeric stealth coatings and are tested for their physiologic stability, photothermal properties, and drug delivery in metastatic triple-negative breast cancer cell (TNBC) lines. Our results indicate that stealth polymer-coated nanoparticles exhibit superior colloidal stability under physiologic conditions, and are excellent photothermal agents, as determined by the increase in temperature of solution in the presence of nanoparticles, upon laser irradiation. The chemotherapeutic drug-loaded nanoparticles also showed concentration-dependent toxicities in TNBC and in a brain metastatic cell line. SIGNIFICANCE STATEMENT: This study develops, for the first time, biocompatible core-shell nanoparticles in a template-free approach that can serve as a drug delivery carrier and as photothermal agents for cancer treatment.

Fast and easy quantitative characterization of methanotroph-photoautotroph cocultures.

Recent research has demonstrated that synthetic methanotroph-photoautotroph cocultures offer a highly promising route to convert biogas into value-added products. However, there is a lack of techniques for fast and accurate characterization of cocultures, such as determining the individual biomass concentration of each organism in real-time. To address this unsolved challenge, we propose an experimental-computational protocol for fast, easy, and accurate quantitative characterization of the methanotroph-photoautotroph cocultures. Besides determining the individual biomass concentration of each organism in the coculture, the protocol can also obtain the individual consumption and production rates of O2 and CO2 for the methanotroph and photoautotroph, respectively. The accuracy and effectiveness of the proposed protocol was demonstrated using two model coculture pairs, Methylomicrobium alcaliphilum 20ZR-Synechococcus sp. PCC7002 that prefers high pH high salt condition, and Methylococcus capsulatus-Chlorella sorokiniana that prefers low salt and neutral pH medium. The performance of the proposed protocol was compared with a flow cytometry-based cell counting approach. The experimental results show that the proposed protocol is much easier to carry out and delivers faster and more accurate results in measuring individual biomass concentration than the cell counting approach without requiring any special equipment.

Protein causes a glycemic response.

The glycemic index is used to compare the extent to which the blood glucose level increases following the consumption of foods containing digestible carbohydrate and is considered to be zero, or not measurable, if the food, such as protein, is carbohydrate-free. We have found that after overnight fasting, the consumption of several varieties of meat caused significant increases in blood glucose levels. We consider these possibly to be because of gluconeogenesis from the digested protein. It is a curious feature that in two instances the response was inversely related to the amount of meat consumed, over the range from 26 to 78 g of protein.

The glycemic response is a personal attribute.

The Glycemic Index (GI) is a measure of the extent of the change in blood glucose content (glycemic response) following consumption of digestible carbohydrate, relative to a standard such as glucose. We have explored whether the reported GIs of foods are a sufficient guide to a person wishing to avoid large glycemic responses and thereby avoid hyperglycemia. For this purpose, volunteers carried out multiple tests of four foods, following overnight fasting, measuring the glycemic response over 2 H. The areas under the blood glucose/time curves (AUCs) were compared. Each food tester displayed individual, characteristic glycemic responses to each food, unrelated to any other tester's response. Wide variations (up to 5-fold) were seen between the average AUCs for the same test by different testers. The absolute magnitudes of the glycemic responses are important for individuals trying to control blood sugar and/or body weight, but using published GI lists as a guide to control the glycemic response is not fully informative. This is because in calculating the GI, individual glycemic responses to glucose are normalized to 100. GI values are, therefore, relative and are not necessarily a reliable guide to the person's actual individual AUC when consuming a food. Without knowledge of the person's characteristic blood glucose responses, reliance only on the GI may be misleading.

Effect of magnetic field alignment of cellulose nanocrystals in starch nanocomposites: Physicochemical and mechanical properties.

The magnetic field (MF) induced alignment of cellulose nanocrystals (CNC) within a starch matrix is investigated and its effect on the physicochemical and mechanical properties of the nanocomposites are discussed in the paper. Two different kinds of CNC i.e. plant-CNC and tunicate-CNC and its hybrid combination are studied to understand the effect of aspect ratio of CNC on the properties of nanocomposite. Nanocomposites with tunicate sourced CNC showed higher tensile strength and modulus, and lower water vapor permeability as compared to plant sourced CNC. These properties are higher for nanocomposites prepared under MF. The modulus of starch nanocomposites increased from 0.26 GPa and 0.32 GPa to 0.38 GPa and 0.44 GPa, respectively for plant-CNC and tunicate-CNC when exposed to MF. The improved orientation and alignment of CNC in presence of MF is further supported by Raman and scanning electron micrographs studies.

Determination of the optical properties of turbid media using relative interstitial radiance measurements: Monte Carlo study, experimental validation, and sensitivity analysis.

Interstitial quantification of the optical properties of tissue is important in biomedicine for both treatment planning of minimally invasive laser therapies and optical spectroscopic characterization of tissues, for example, prostate cancer. In a previous study, we analyzed a method first demonstrated by Dickey et al., [Phys. Med. Biol. 46, 2359 (2001)] to utilize relative interstitial steady-state radiance measurements for recovering the optical properties of turbid media. The uniqueness of point radiance measurements were demonstrated in a forward sense, and strategies were suggested for improving performance under noisy experimental conditions. In this work, we test our previous conclusions by fitting the P3 approximation for radiance to Monte Carlo predictions and experimental data in tissue-simulating phantoms. Fits are performed at: 1. a single sensor position (0.5 or 1 cm), 2. two sensor positions (0.5 and 1 cm), and 3. a single sensor position (0.5 or 1 cm) with input knowledge of the sample's effective attenuation coefficient. The results demonstrate that single sensor radiance measurements can be used to retrieve optical properties to within approximately 20%, provided the transport albedo is greater than approximately 0.9. Furthermore, compared to the single sensor fits, employing radiance data at two sensor positions did not significantly improve the accuracy of recovered optical properties. However, with knowledge of the effective attenuation coefficient of the medium, optical properties can be retrieved experimentally to within approximately 10% for an albedo greater or equal to 0.5.

Perturbative diffusion theory formalism for interpreting temporal light intensity changes during laser interstitial thermal therapy.

In an effort to understand dynamic optical changes during laser interstitial thermal therapy (LITT), we utilize the perturbative solution of the diffusion equation in heterogeneous media to formulate scattering weight functions for cylindrical line sources. The analysis explicitly shows how changes in detected interstitial light intensity are associated with the extent and location of the volume of thermal coagulation during treatment. Explanations for previously reported increases in optical intensity observed early during laser heating are clarified using the model and demonstrated with experimental measurements in ex vivo bovine liver tissue. This work provides an improved understanding of interstitial optical signal changes during LITT and indicates the sensitivity and potential of interstitial optical monitoring of thermal damage.

Why the linkage of glycogen to glycogenin was so difficult to determine.

Glycogenin is the self-glucosylating enzyme that primes mammalian and yeast glycogen synthesis. It proved to be the long-suspected, covalently bound protein component of glycogen. One of the most difficult aspects in elucidating the role of glycogenin was to learn the nature of its covalent bond to glycogen.

Glycogenin: the primer for mammalian and yeast glycogen synthesis.

Glycogen synthesis, whether in mammalian tissue, yeast, or Agrobacterium tumefaciens or other bacteria, is initiated by autoglucosylation of a protein. Initiation in muscle, by a self-glucosylating protein, glycogenin-1, is the most thoroughly studied system, as is described here. These relatively recent findings have prompted a rekindling of interest in the intermediates lying between the primer and mature mammalian glycogen.

Optical and acoustic properties at 1064 nm of polyvinyl chloride-plastisol for use as a tissue phantom in biomedical optoacoustics.

A novel optoacoustic phantom made of polyvinyl chloride-plastisol (PVCP) for optoacoustic studies is described. The optical and acoustic properties of PVCP were measured. Titanium dioxide (TiO2) powder and black plastic colour (BPC) were used to introduce scattering and absorption, respectively, in the phantoms. The optical absorption coefficient (mua) at 1064 nm was determined using an optoacoustic method, while diffuse reflectance measurements were used to obtain the optical reduced scattering coefficient (mu's). These optical properties were calculated to be mua = (12.818 +/- 0.001)ABPC cm(-1) and mu's = (2.6 +/- 0.2)S(TiO2) + (1.4 +/- 0.1) cm(-1), where ABPC is the BPC per cent volume concentration, and S(TiO2) is the TiO2 volume concentration (mg mL(-1)). The speed of sound in PVCP was measured to be (1.40 +/- 0.02) x 10(3) m s(-1) using the pulse echo transmit receive method, with an acoustic attenuation of (0.56 +/- 1.01) f(1.51+/-0.06)MHz (dB cm(-1)) in the frequency range of 0.61-1.25 MHz, and a density, calculated by measuring the displacement of water, of 1.00 +/- 0.04 g cm(-3). The speed of sound and density of PVCP are similar to tissue, and together with the user-adjustable optical properties, make this material well suited for developing tissue-equivalent phantoms for biomedical optoacoustics.

Laser photothermoacoustic heterodyned lock-in depth profilometry in turbid tissue phantoms.

Frequency-domain correlation and spectral analysis photothermoacoustic (FD-PTA) imaging is a promising new technique, which is being developed to detect tumor masses in turbid biological tissue. Unlike conventional biomedical photoacoustics which uses time-of-flight acoustic information induced by a pulsed laser to indicate the tumor size and location, in this research, a new FD-PTA instrument featuring frequency sweep (chirp) and heterodyne modulation and lock-in detection of a continuous-wave laser source at wavelength is constructed and tested for its depth profilometric capabilities with regard to turbid media imaging. Owing to the linear relationship between the depth of acoustic signal generation and the delay time of signal arrival to the transducer, information specific to a particular depth can be associated with a particular frequency in the chirp signal. Scanning laser-fluence modulation frequencies with a linear frequency sweep method preserves the depth-to-delay time linearity and recovers FD-PTA signals from a range of depths. Combining with the depth information carried by the back-propagated acoustic chirp signal at each scanning position, one could rapidly generate subsurface three-dimensional images of the scanning area at optimal signal-to-noise ratios and low laser fluences, a combination of tasks that is difficult or impossible by use of pulsed photoacoustic detection. In this paper, results of PTA scans performed on tissue mimicking control phantoms with various optical, acoustical, and geometrical properties are presented. A mathematical model is developed to study the laser-induced photothermoacoustic waves in turbid media. The model includes both the scattering and absorption properties of the turbid medium. A good agreement is obtained between the experimental and numerical results. It is concluded that frequency domain photothermoacoustics using a linear frequency sweep method and heterodyne lock-in detection has the potential to be a reliable tool for biomedical depth-profilometric imaging.

Differential scanning calorimetry of whole Escherichia coli treated with the antimicrobial peptide MSI-78 indicate a multi-hit mechanism with ribosomes as a novel target.

Differential Scanning Calorimetry (DSC) of intact Escherichia coli (E. coli) was used to identify non-lipidic targets of the antimicrobial peptide (AMP) MSI-78. The DSC thermograms revealed that, in addition to its known lytic properties, MSI-78 also has a striking effect on ribosomes. MSI-78's effect on DSC scans of bacteria was similar to that of kanamycin, an antibiotic drug known to target the 30S small ribosomal subunit. An in vitro transcription/translation assay helped confirm MSI-78's targeting of ribosomes. The scrambled version of MSI-78 also affected the ribosome peak of the DSC scans, but required greater amounts of peptide to cause a similar effect to the unscrambled peptide. Furthermore, the effect of the scrambled peptide was not specific to the ribosomes; other regions of the DSC thermogram were also affected. These results suggest that MSI-78's effects on E. coli are at least somewhat dependent on its particular structural features, rather than a sole function of its overall charge and hydrophobicity. When considered along with earlier work detailing MSI-78's membrane lytic properties, it appears that MSI-78 operates via a multi-hit mechanism with multiple targets.

Interstitial diffuse radiance spectroscopy of gold nanocages and nanorods in bulk muscle tissues.

Radiance spectroscopy was applied to the interstitial detection of localized inclusions containing Au nanocages or nanorods with various concentrations embedded in porcine muscle phantoms. The radiance was quantified using a perturbation approach, which enabled the separation of contributions from the porcine phantom and the localized inclusion, with the inclusion serving as a perturbation probe of photon distributions in the turbid medium. Positioning the inclusion at various places in the phantom allowed for tracking of photons that originated from a light source, passed through the inclusion's location, and reached a detector. The inclusions with high extinction coefficients were able to absorb nearly all photons in the range of 650-900 nm, leading to a spectrally flat radiance signal. This signal could be converted to the relative density of photons incident on the inclusion. Finally, the experimentally measured quantities were expressed via the relative perturbation and arranged into the classical Beer-Lambert law that allowed one to extract the extinction coefficients of various types of Au nanoparticles in both the transmission and back reflection geometries. It was shown that the spatial variation of perturbation could be described as 1/r dependence, where r is the distance between the inclusion and the detector. Due to a larger absorption cross section, Au nanocages produced greater perturbations than Au nanorods of equal particle concentration, indicating a better suitability of Au nanocages as contrast agents for optical measurements in turbid media. Individual measurements from different inclusions were combined into detectability maps.

Models and measurements of light intensity changes during laser interstitial thermal therapy: implications for optical monitoring of the coagulation boundary location.

We have developed a multi-region spherical Monte Carlo (MC) model to simulate the dynamic changes in light intensity measured during laser interstitial thermal therapy (LITT). Model predictions were validated experimentally in tissue-simulating albumen phantoms with well-characterized optical properties that vary dynamically with LITT in a way similar to tissue. For long treatments (2.5 W, approximately 1800 s), the transient light intensity changes demonstrated better qualitative agreement with a three-region MC model (with an inner layer of fully coagulated optical properties, a middle layer of partially coagulated properties and an outer region of native properties); for short treatments (4 W, approximately 240 s), better qualitative agreement was seen with a two-region MC model (with an inner layer of fully coagulated properties and outer region of native properties). These differences were attributed to differences in coagulation formation during low- and high-powered heating regimes, respectively. At the end of heating, a three-region coagulation zone was observed for both heating schemes. Quantitatively, final light intensity changes at the end of heating were compared with changes predicted by both two- and three-region MC for the same experimentally measured coagulation size and found to agree within approximately 30% for both models. The developed MC model helps lend insight into the nature of thermal coagulation events occurring for low and high power LITT irradiation schemes.

Feasibility of interstitial near-infrared radiance spectroscopy platform for ex vivo canine prostate studies: optical properties extraction, hemoglobin and water concentration, and gold nanoparticles detection.

The canine prostate is a close match for the human prostate and is used in research of prostate cancers. Determining accurately optical absorption and scattering properties of the gland in a wide spectral range (preferably in a minimally invasive way), linking optical properties to concentrations of major endogenous chromophores, and detecting the presence of localized optical inhomogeneities like inclusions of gold nanoparticles for therapeutic and diagnostic purposes, are among the major challenges for researchers. The goal of the article is to demonstrate a feasibility of the multifunctional radiance spectroscopy platform in providing the required information. For ex vivo canine prostate, extraction of the effective attenuation and diffusion coefficients using relative cw radiance measurements was demonstrated in the 650- to 900-nm range. The derived absorption coefficient was decomposed to contributions from 9.0 µM HbO2, 29.6 µM Hb, and 0.47 fractional volume of H2O. Detection of a localized inclusion containing ∼1.5·1010 gold nanorods (0.8 µg Au) at 10 mm distance from the urethra was achieved with the detector in the urethra and the light source in a virtual rectum position. The platform offers the framework for a systematic study of various chromophores in the prostate that can be used as comprehensive diagnostic markers.

Optical absorption and scattering properties of bulk porcine muscle phantoms from interstitial radiance measurements in 650-900 nm range.

We demonstrated the application of relative radiance-based continuous wave (cw) measurements for recovering absorption and scattering properties (the effective attenuation coefficient, the diffusion coefficient, the absorption coefficient and the reduced scattering coefficient) of bulk porcine muscle phantoms in the 650-900 nm spectral range. Both the side-firing fiber (the detector) and the fiber with a spherical diffuser at the end (the source) were inserted interstitially at predetermined locations in the phantom. The porcine phantoms were prostate-shaped with ∼4 cm in diameter and ∼3 cm thickness and made from porcine loin or tenderloin muscles. The described method was previously validated using the diffusion approximation on simulated and experimental radiance data obtained for homogenous Intralipid-1% liquid phantom. The approach required performing measurements in two locations in the tissue with different distances to the source. Measurements were performed on 21 porcine phantoms. Spectral dependences of the effective attenuation and absorption coefficients for the loin phantom deviated from corresponding dependences for the tenderloin phantom for wavelengths <750 nm. The diffusion constant and the reduced scattering coefficient were very close for both phantom types. To quantify chromophore presence, the plot for the absorption coefficient was matched with a synthetic absorption spectrum constructed from deoxyhemoglobin, oxyhemoglobin and water. The closest match for the porcine loin spectrum was obtained with the following concentrations: 15.5 µM (±30% s.d.) Hb, 21 µM (±30% s.d.) HbO2 and 0.3 (±30% s.d.) fractional volume of water. The tenderloin absorption spectrum was best described by 30 µM Hb (±30% s.d), 19 µM (±30% s.d.) HbO2 and 0.3 (±30% s.d.) fractional volume of water. The higher concentration of Hb in tenderloin was consistent with a dark-red appearance of the tenderloin phantom. The method can be applied to a number of biological tissues and organs for interstitial optical interrogation.