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1.
Front Bioeng Biotechnol ; 11: 1191327, 2023.
Article in English | MEDLINE | ID: mdl-37545884

ABSTRACT

The new and unique possibilities that nanomaterials offer have greatly impacted biomedicine, from the treatment and diagnosis of diseases, to the specific and optimized delivery of therapeutic agents. Technological advances in the synthesis, characterization, standardization, and therapeutic performance of nanoparticles have enabled the approval of several nanomedicines and novel applications. Discoveries continue to rise exponentially in all disease areas, from cancer to neurodegenerative diseases. In Spain, there is a substantial net of researchers involved in the development of nanodiagnostics and nanomedicines. In this review, we summarize the state of the art of nanotechnology, focusing on nanoparticles, for the treatment of diseases in Spain (2017-2022), and give a perspective on the future trends and direction that nanomedicine research is taking.

2.
Nanoscale ; 15(28): 12124, 2023 Jul 20.
Article in English | MEDLINE | ID: mdl-37435809

ABSTRACT

Correction for 'Label free localization of nanoparticles in live cancer cells using spectroscopic microscopy' by Graham L. C. Spicer et al., Nanoscale, 2018, 10, 19125-19130, https://doi.org/10.1039/C8NR07481J.

3.
J Biophotonics ; 14(2): e202000341, 2021 02.
Article in English | MEDLINE | ID: mdl-33128802

ABSTRACT

Temperature measurement at the nanoscale has brought insight to a wide array of research interests in modern chemistry, physics, and biology. These measurements have been enabled by the advent of nanothermometers, which relay nanoscale temperature information through the analysis of their intrinsic photophysical behavior. In the past decade, several nanothermometers have been developed including dyes, nanodiamonds, fluorescent proteins, nucleotides, and nanoparticles. However, temperature measurement using intact DNA has not yet been achieved. Here, we present a method to study the temperature sensitivity of the DNA molecule within a physiologic temperature range when complexed with fluorescent dye. We theoretically and experimentally report the temperature sensitivity of the DNA-Hoechst 33342 complex in different sizes of double-stranded oligonucleotides and plasmids, showing its potential use as a nanothermometer. These findings allow for extending the thermal study of DNA to several research fields including DNA nanotechnology, optical tweezers, and DNA nanoparticles.


Subject(s)
Nanodiamonds , Nanotechnology , DNA , Fluorescent Dyes , Temperature
4.
J Biophotonics ; 12(9): e201900044, 2019 09.
Article in English | MEDLINE | ID: mdl-31034763

ABSTRACT

In the last decade, technological advances in chemistry and photonics have enabled real-time measurement of temperature at the nanoscale. Nanothermometers, probes specifically designed to relay these nanoscale temperature changes, provide a high degree of temperature, temporal, and spatial resolution and precision. Several different approaches have been proposed, including microthermocouples, luminescence and fluorescence polarization anisotropy-based nanothermometers. Anisotropy-based nanothermometers excel in terms of biocompatibility because they can be built from endogenous proteins conjugated to dyes, minimizing any system perturbation. Moreover, the resulting fluorescent proteins can retain their native structure and activity while performing the temperature measurement, allowing precise temperature recordings from the native environment or during an enzymatic reaction in any given experimental system. To facilitate the future use of these nanothermometers in research, here we present a theoretical model that predicts the optimal sensitivity for anisotropy-based thermometers starting with any protein or dye, based on protein size and dye fluorescence lifetime. Using this model, most proteins and dyes can be converted to nanothermometers. The utilization of these nanothermometers by a broad spectrum of disciplines within the scientific community will bring new knowledge and understanding that today remains unavailable with current techniques.


Subject(s)
Coloring Agents/chemistry , Nanotechnology/methods , Proteins/chemistry , Thermometry/methods , Animals , Anisotropy , Biocompatible Materials , Cattle , Chickens , Fluorescent Dyes/chemistry , Fluorescent Dyes/pharmacology , Horses , Light , Luminescence , Nanoparticles/chemistry , Temperature
5.
Adv Exp Med Biol ; 1106: 95-108, 2018.
Article in English | MEDLINE | ID: mdl-30484155

ABSTRACT

Almost 15 years ago, the URI prefoldin-like complex was discovered by Krek and colleagues in immunoprecipitation experiments conducted in mammalian cells with the aim of identifying new binding partners of the E3 ubiquitin-protein ligase S-phase kinase-associated protein 2 (SKP2) (Gstaiger et al. Science 302(5648):1208-1212, 2003). The URI prefoldin-like complex is a heterohexameric chaperone complex comprising two α and four ß subunits (α2ß4). The α subunits are URI and STAP1, while the ß subunits are PFDN2, PFDN6, and PFDN4r, one of which is probably present in duplicate. Elucidating the roles and functions of these components in vitro and in vivo will help to clarify the mechanistic behavior of what appears to be a remarkably important cellular machine.


Subject(s)
Molecular Chaperones/chemistry , Animals , Immunoprecipitation
6.
Nanoscale ; 10(40): 19125-19130, 2018 Oct 18.
Article in English | MEDLINE | ID: mdl-30298892

ABSTRACT

Gold nanoparticles (GNPs) have become essential tools used in nanobiotechnology due to their tunable plasmonic properties and low toxicity in biological samples. Among the available approaches for imaging GNPs internalized by cells, hyperspectral techniques stand out due to their ability to simultaneously image and perform spectral analysis of GNPs. Here, we present a study utilizing a recently introduced hyperspectral imaging technique, live-cell PWS, for the imaging, tracking, and spectral analysis of GNPs in live cancer cells. Using principal components analysis, the extracellular or intracellular localization of the GNPs can be determined without the use of exogenous labels. This technique uses wide-field white light, assuring minimal toxicity and suitable signal-to-noise ratio for spectral and temporal resolution of backscattered signal from GNPs and local cellular structures. The application of live-cell PWS introduced here could make a great impact in nanomedicine and nanotechnology by giving new insights into GNP internalization and intracellular trafficking.


Subject(s)
Gold , Materials Testing , Metal Nanoparticles/chemistry , Neoplasms/diagnostic imaging , Gold/chemistry , Gold/pharmacology , HeLa Cells , Humans , Neoplasms/metabolism
7.
Bioorg Med Chem ; 26(18): 5224-5228, 2018 10 01.
Article in English | MEDLINE | ID: mdl-30262133

ABSTRACT

Photodynamic therapy (PDT) is a non-invasive treatment widely applied to different cancers. The goal of PDT is the photo-induced destruction of cancer cells by the activation of different cell death mechanisms, including apoptosis and/or necrosis. Recent efforts focusing on understanding the mechanisms of cell death activated by PDT find that it depends on the type of photosensitizer (PS), targeted organelles, and nature of the light used. It is generally accepted that very short incubation times are required to direct the PS to the plasma membrane (PM), while longer periods result in the accumulation of the PS in internal compartments such as the endoplasmic reticulum or mitochondria. Glycosylation of the PS targets cancer via saccharide receptors on the cell surface, and is generally assumed that these compounds rapidly internalize and accumulate, e.g. in the endoplasmic reticulum. Herein we demonstrate that a minor fraction of a glycosylated chlorin compound residing at the PM of cancer cells can activate necrosis upon illumination by compromising the PM independently of the length of the incubation period. The results presented here show that the PM can also be targeted by glycosylated PS designed to accumulate in internal organelles. PS activation to induce necrosis by compromising the plasma membrane has the benefits of fast cell death and shorter irradiation times. The findings described here expand our understanding of the cellular damage induced by phototherapies, presenting the possibility of activating another cell death mechanism based on the incubation time and type of light used.


Subject(s)
Cell Membrane/drug effects , Necrosis/drug therapy , Photochemotherapy , Photosensitizing Agents/pharmacology , Animals , CHO Cells , Cell Death/drug effects , Cell Membrane/metabolism , Cells, Cultured , Cricetulus , Dose-Response Relationship, Drug , Molecular Structure , Necrosis/metabolism , Photosensitizing Agents/chemistry , Structure-Activity Relationship
8.
Nanoscale ; 9(17): 5404-5407, 2017 May 04.
Article in English | MEDLINE | ID: mdl-28426045

ABSTRACT

A method is proposed for controlling the number of nanoparticles bound to cell membranes via RGDS peptide-integrin interactions. It consists of propelling nanoparticles bearing the peptides with enzymes (glucose oxidase), which disrupts biomolecular interactions as a function of the concentration of enzyme substrate (glucose).


Subject(s)
Enzymes, Immobilized/chemistry , Glucose Oxidase/chemistry , Glucose/chemistry , Nanoparticles , Receptors, Cell Surface/metabolism , Animals , CHO Cells , Cricetulus
9.
Small ; 13(15)2017 04.
Article in English | MEDLINE | ID: mdl-28151578

ABSTRACT

A universal method for inactivating enzymes on demand is introduced, which involves irradiating nanorod-bound enzymes with near-infrared light. The subsequent generation of plasmonic heat denatures the enzymes selectively without damaging other proteins or cell membranes present in the same solution.


Subject(s)
Glucose Oxidase/metabolism , Horseradish Peroxidase/metabolism , Light , Nanotubes/chemistry , Temperature , Animals , CHO Cells , Cricetinae , Cricetulus , Enzyme Activation/radiation effects
10.
Opt Lett ; 40(21): 4931-4, 2015 Nov 01.
Article in English | MEDLINE | ID: mdl-26512486

ABSTRACT

We previously established that spectroscopic microscopy can quantify subdiffraction-scale refractive index (RI) fluctuations in a label-free dielectric medium with a smooth surface. However, to study more realistic samples, such as biological cells, the effect of rough surface should be considered. In this Letter, we first report an analytical theory to synthesize microscopic images of a rough surface, validate this theory by finite-difference time-domain (FDTD) solutions of Maxwell's equations, and characterize the spectral properties of light reflected from a rough surface. Then, we report a technique to quantify the RI fluctuations beneath a rough surface and demonstrate its efficacy on FDTD-synthesized spectroscopic microscopy images, as well as experimental data obtained from biological cells.


Subject(s)
Microscopy/methods , Mouth Mucosa/cytology , Mouth Mucosa/physiology , Nephelometry and Turbidimetry/methods , Refractometry/methods , Spectrum Analysis/methods , Algorithms , Cells, Cultured , Computer Simulation , Humans , Models, Biological , Models, Statistical , Reproducibility of Results , Sensitivity and Specificity , Surface Properties
11.
Photochem Photobiol ; 90(2): 419-30, 2014.
Article in English | MEDLINE | ID: mdl-24112086

ABSTRACT

The photophysical properties of a chlorin, isobacteriochlorin and bacteriochlorin built on a core tetrapentafluorophenylporphyrin (TPPF20 ) and the nonhydrolyzable para thioglycosylated conjugates of these chromophores are presented. The photophysical characterization of these compounds was done in three different solvents to correlate with different environments in cells and tissues. Compared with TPPF20 other dyes have greater absorption in the red region of the visible spectrum and greater fluorescence quantum yields. The excited state lifetimes are from 3 to 11 ns. The radiative and nonradiative rate constants for deactivation of the excited state were estimated from the fluorescence quantum yield and excited state lifetime. The data indicate that the bacteriochlorin has strong absorption bands near 730 nm and efficiently enters the triplet manifold. The isobacteriochlorin has a 40-70% fluorescence quantum yield depending on solvent, so it may be a good fluorescent tag. The isobacteriochlorins also display enhanced two-photon absorption, thereby allowing the use of 860 nm light to excite the compound. While the two-photon cross section of 25 GM units is not large, excitation of low chromophore concentrations can induce apoptosis. The glycosylated compounds accumulate in cancer cells and a head and neck squamous carcinoma xenograft tumor model in mice. These compounds are robust to photobleaching.


Subject(s)
Photosensitizing Agents/chemistry , Porphyrins/chemistry , Animals , Glycosylation , Mice , NIH 3T3 Cells , Photons , Photosensitizing Agents/therapeutic use , Porphyrins/therapeutic use , Spectrometry, Fluorescence , Spectrophotometry, Ultraviolet
12.
ACS Nano ; 7(10): 8666-72, 2013 Oct 22.
Article in English | MEDLINE | ID: mdl-24047507

ABSTRACT

Controlling and monitoring temperature at the single cell level has become pivotal in biology and medicine. Indeed, temperature influences many intracellular processes and is also involved as an activator in novel therapies. Aiming to assist such developments, several approaches have recently been proposed to probe cell temperature in vitro. None of them have so far been extended to a living organism. Here we present the first in vivo intracellular temperature imaging. Our technique relies on measuring the fluorescence polarization anisotropy of green fluorescent protein (GFP) on a set of GFP expressing neurons in Caenorhabditis elegans (C. elegans). We demonstrate fast and noninvasive monitoring of subdegree temperature changes on a single neuron induced by local photoheating of gold nanoparticles. This simple and biocompatible technique is envisioned to benefit several fields including hyperthermia treatment, selective drug delivery, thermal regulation of gene expression and neuron laser ablation.


Subject(s)
Caenorhabditis elegans/physiology , Hot Temperature , Animals , Caenorhabditis elegans/genetics , Fluorescence Polarization , Gold/chemistry , Green Fluorescent Proteins/genetics , Metal Nanoparticles
13.
Nano Lett ; 12(4): 2107-11, 2012 Apr 11.
Article in English | MEDLINE | ID: mdl-22394124

ABSTRACT

Heat is of fundamental importance in many cellular processes such as cell metabolism, cell division and gene expression. (1-3) Accurate and noninvasive monitoring of temperature changes in individual cells could thus help clarify intricate cellular processes and develop new applications in biology and medicine. Here we report the use of green fluorescent proteins (GFP) as thermal nanoprobes suited for intracellular temperature mapping. Temperature probing is achieved by monitoring the fluorescence polarization anisotropy of GFP. The method is tested on GFP-transfected HeLa and U-87 MG cancer cell lines where we monitored the heat delivery by photothermal heating of gold nanorods surrounding the cells. A spatial resolution of 300 nm and a temperature accuracy of about 0.4 °C are achieved. Benefiting from its full compatibility with widely used GFP-transfected cells, this approach provides a noninvasive tool for fundamental and applied research in areas ranging from molecular biology to therapeutic and diagnostic studies.


Subject(s)
Gold/chemistry , Green Fluorescent Proteins/chemistry , Metal Nanoparticles/chemistry , Temperature , Cell Line, Tumor , Green Fluorescent Proteins/metabolism , HeLa Cells , Humans , Particle Size , Photochemical Processes , Surface Properties
14.
Bioconjug Chem ; 21(11): 2136-46, 2010 Nov 17.
Article in English | MEDLINE | ID: mdl-20964323

ABSTRACT

The facile synthesis and photophysical properties of three nonhydrolyzable thioglycosylated porphyrinoids are reported. Starting from meso-perfluorophenylporphyrin, the nonhydrolyzable thioglycosylated porphyrin (PGlc4), chlorin (CGlc4), isobacteriochlorin (IGlc4), and bacteriochlorin (BGlc4) can be made in 2-3 steps. The ability to append a wide range of targeting agents onto the perfluorophenyl moieties, the chemical stability, and the ability to fine-tune the photophysical properties of the chromophores make this a suitable platform for development of biochemical tags, diagnostics, or as photodynamic therapeutic agents. Compared to the porphyrin in phosphate buffered saline, CGlc4 has a markedly greater absorbance of red light near 650 nm and a 6-fold increase in fluorescence quantum yield, whereas IGlc4 has broad Q-bands and a 12-fold increase in fluorescence quantum yield. BGlc4 has a similar fluorescence quantum yield to PGlc4 (<10%), but the lowest-energy absorption/emission peaks of BGlc4 are considerably red-shifted to near 730 nm with a nearly 50-fold greater absorbance, which may allow this conjugate to be an effective PDT agent. The uptake of CGlc4, IGlc4, and BGlc4 derivatives into cells such as human breast cancer cells MDA-MB-231 and K:Molv NIH 3T3 mouse fibroblast cells can be observed at nanomolar concentrations. Photobleaching under these conditions is minimal.


Subject(s)
Molecular Imaging/methods , Porphyrins/chemistry , Sulfhydryl Compounds/chemistry , Animals , Cell Line, Tumor , Fluorescence , Humans , Mice , Molecular Structure , NIH 3T3 Cells , Photochemistry , Porphyrins/chemical synthesis , Stereoisomerism , Sulfhydryl Compounds/chemical synthesis
15.
Cancer Lett ; 299(1): 72-9, 2010 Dec 18.
Article in English | MEDLINE | ID: mdl-20805015

ABSTRACT

A characteristic of cancer cells is the generation of lactate from glucose in spite of adequate oxygen for oxidative phosphorylation. This property - known as the "Warburg effect" or aerobic glycolysis - contrasts with anaerobic glycolysis, which is triggered in hypoxic normal cells. The Warburg effect is thought to provide a means for cancer cells to survive under conditions where oxygen is limited and to generate metabolites necessary for cell growth. The shift from oxidative phosphorylation to glycolysis in response to hypoxia is mediated by the production of hypoxia-inducible factor (HIF) - a transcription factor family that stimulates the expression of proteins involved in glucose uptake and glycolysis. We reported previously that elevated phospholipase D (PLD) activity in renal and breast cancer cells is required for the expression of the α subunits of HIF1 and HIF2. We report here that the aerobic glycolysis observed in human breast and renal cancer cells is dependent on the elevated PLD activity. Intriguingly, the effect of PLD on the Warburg phenotype was dependent on the mammalian target of rapamycin complex 1 (mTORC1) in the breast cancer cells and on mTORC2 in the renal cancer cells. These data indicate that elevated PLD-mTOR signaling, which is common in human cancer cells, is critical for the metabolic shift to aerobic glycolysis.


Subject(s)
Glucose/metabolism , Glycolysis , Intracellular Signaling Peptides and Proteins/physiology , Neoplasms/metabolism , Phospholipase D/physiology , Protein Serine-Threonine Kinases/physiology , Cell Line, Tumor , Glucose Transport Proteins, Facilitative/analysis , Humans , Mechanistic Target of Rapamycin Complex 1 , Multiprotein Complexes , Oxidative Phosphorylation , Phospholipase D/antagonists & inhibitors , Proteins , TOR Serine-Threonine Kinases , Transcription Factors/physiology
16.
Anal Chem ; 81(24): 10167-71, 2009 Dec 15.
Article in English | MEDLINE | ID: mdl-19911810

ABSTRACT

While cancer is still an implacable disease, many cancers can be cured if they are diagnosed in an early stage. Recently, it was reported that the transformation from normal cells to cancer cells can change their mechanoelastic properties to become softer and more deformable. If some cancer cells are more deformable, then a progressive increase of the volume of softer cancer cells should be induced as an abrupt change in osmolarity is applied. On the basis of this hypothesis, we developed a sensor that can electronically monitor the volume increase of cancer cells under hyposmotic pressure. By this methodology, K:Molv NIH 3T3 cells, 786-O human kidney carcinoma cells, and MPSC-1 ovarian cancer cells were successfully detected within 30 min using on the order of 10 cells. These cancer cells could be detected with the same sensitivity even in the presence of a vast excess of the respective noncancerous cells [NIH 3T3 cells, human embryonic kidney (HEK) 293 cells, ovarian surface epithelial (OSE) cells]. Since the proposed impedimetric sensor could be useful for detecting cancer cells fast and reliably, it could be further implemented in the screening of large populations of tissue samples and the detection of circulating tumor cells for point-of-care applications.


Subject(s)
Carcinoma, Renal Cell/pathology , Cell Separation/instrumentation , Cell Separation/methods , Kidney Neoplasms/pathology , Ovarian Neoplasms/pathology , Transducers , Animals , Cell Line, Tumor , Female , Humans , Mice , NIH 3T3 Cells , Sensitivity and Specificity
17.
Photochem Photobiol Sci ; 7(11): 1415-21, 2008 Nov.
Article in English | MEDLINE | ID: mdl-18958330

ABSTRACT

A water-soluble tetra-S-glycosylated porphyrin (P-Glu(4)) is absorbed by MDA-MB-231 human breast cancer cells whereupon irradiation with visible light causes necrosis or apoptosis depending on the concentration of the porphyrin and the power of the light. With the same amount of light irradiation power (9.4 W m(-2)), at 10-20 microM concentrations necrosis is predominantly observed, while at <10 microM concentrations, apoptosis is the principal cause of cell death. Of the various possible pathways for the induction of apoptosis, experiments demonstrate that calcium is released from the endoplasmic reticulum, cytochrome c is liberated from the mitochondria to the cytosol, pro-caspase-3 is activated, poly-(ADP-ribose) polymerase is cleaved, and the chromatin is condensed subsequent to photodynamic treatment of these cells. Confocal microscopy indicates a substantial portion of the P-Glu(4) is located in the endoplasmic reticulum at <10 microM. These data indicate that the photodynamic treatment of MDA-MB-231 cells using low concentrations of the P-Glu(4) porphyrin and low light induces apoptosis mostly initiated from stress produced to the endoplasmic reticulum.


Subject(s)
Apoptosis/drug effects , Apoptosis/radiation effects , Breast Neoplasms/pathology , Endoplasmic Reticulum/metabolism , Light , Porphyrins/chemistry , Porphyrins/pharmacology , Animals , Calcium/metabolism , Caspases/metabolism , Cattle , Cell Line, Tumor , Cytochromes c/metabolism , Endoplasmic Reticulum/drug effects , Endoplasmic Reticulum/radiation effects , Enzyme Activation/drug effects , Glycosylation , Hydrolysis , Indoles , Microscopy, Confocal , Microscopy, Fluorescence , Mitochondria/metabolism , Poly(ADP-ribose) Polymerases/metabolism , Staining and Labeling , Stress, Physiological/drug effects , Stress, Physiological/radiation effects
18.
Appl Spectrosc ; 58(3): 344-8, 2004 Mar.
Article in English | MEDLINE | ID: mdl-15035717

ABSTRACT

A spectroscopic device for monitoring the temperature of aqueous solutions is presented. It uses a 950 nm light emission diode as light source and two photodiodes as detectors. Temperature is monitored following the thermally induced absorbance changes of the water-OH second overtone (approximately 960 nm). A linear response between the light absorbed by an aqueous solution and its temperature is found in the range from 15 to 95 degrees C. A prediction error of 0.1 degrees C and a precision of 0.07 degrees C in temperature measurement can be achieved. Up to 0.1 M of electrolyte concentration can be present in the solution without significantly affecting the temperature measurement. Different strategies, such as remote (noninvasive) or in situ (using a fiber-optic probe) temperature measurement, are shown, and their relative advantages are discussed.

19.
Cancer ; 98(2): 262-8, 2003 Jul 15.
Article in English | MEDLINE | ID: mdl-12872343

ABSTRACT

BACKGROUND: Cathepsin B (CB) is a lysosomal cysteine proteinase synthesized as a zymogen of 39-47 kilodaltons (kD), which is subsequently converted into an active single- chain form of 33 kD (CB33) and, by additional processing, into the active 2-chain form containing a heavy chain of 27-29 kD (CB(27-29)) and a light chain of 4-6 kD. Increased or altered CB expression has been documented in a variety of tumor cells, but to the authors' knowledge only one study published to date has reported clinicopathologic significance for CB in transitional cell carcinoma (TCC) of the bladder. METHODS: In this work, CB expression was determined by Western blot analysis in TCC bladder tissue from 30 patients. Nontumor bladder tissue was also analyzed for CB expression. RESULTS: The study results demonstrate higher expression of CB in TCC invasive tumors than in superficial bladder carcinoma. Furthermore, whereas normal bladder only expressed the 29-kD CB protein, tumor and peritumoral tissue demonstrated the 27- to 29-kD CB form. Immunohistochemical staining did not evidence changes in CB localization between tumor and nontumor tissue. CONCLUSIONS: According to the results of the current study, bladder tumor progression appears to be associated with quantitative changes in CB protein expression, as well as with qualitative changes related to the type of CB expressed.


Subject(s)
Biomarkers, Tumor/metabolism , Carcinoma, Transitional Cell/metabolism , Carcinoma, Transitional Cell/pathology , Cathepsin B/metabolism , Urinary Bladder Neoplasms/metabolism , Urinary Bladder Neoplasms/pathology , Adult , Aged , Aged, 80 and over , Blotting, Western , Female , Humans , Immunohistochemistry , Male , Middle Aged , Neoplasm Invasiveness
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