Radiation Cleaved Drug-Conjugate Linkers Enable Local Payload Release.

Conjugation of therapeutic payloads to biologics including antibodies and albumin can enhance the selectively of drug delivery to solid tumors. However, achieving activity in tumors while avoiding healthy tissues remains a challenge, and payload activity in off-target tissues can cause toxicity for many such drug-conjugates. Here, we address this issue by presenting a drug-conjugate linker strategy that releases an active therapeutic payload upon exposure to ionizing radiation. Localized X-ray irradiation at clinically relevant doses (8 Gy) yields 50% drug (doxorubicin or monomethyl auristatin E, MMAE) release under hypoxic conditions that are traditionally associated with radiotherapy resistance. As proof-of-principle, we apply the approach to antibody- and albumin-drug conjugates and achieve >2000-fold enhanced MMAE cytotoxicity upon irradiation. Overall, this work establishes ionizing radiation as a strategy for spatially localized cancer drug delivery.

Highly fluorescent few atoms silver nanoclusters with strong photocatalytic activity synthesized by ultrashort light pulses.

While there are conventional chemical synthesis methods to generate metal nanoclusters (NCs), many of them are adversely affected by the unavoidable contamination of the nanoproduct solution, resulting in aggregation, background noise in analytical chemistry, toxicity, and deactivation of the catalyst. In this work, physical method of ultrafast laser ablation as a "green" synthesis approach together with mechanical centrifugation to obtain silver NCs, simplifying widely the chemical synthesis requirements, is proposed. Remarkably, compared with conventional methods for synthesizing Ag NCs, this new approach starts with a colloid that contains nanosized particles as well as smaller species, managing to obtain colloids with few atoms NCs by centrifugation. Those colloids were analyzed by fluorescence spectroscopy observing UV bands corresponding with HOMO-LUMO cluster transitions. Besides, independent HRTEM measurements were made confirming the presence of few atoms Ag NCs, as well as small NPs in different formation stages. Equally important, photocatalytic efficiency of the obtained NCs was studied through degradation of Methylene Blue (MB) when it was mixed with as-prepared or highly centrifuged colloid, showing an enhanced photocatalytic efficiency of 79% as compared to 57% for pure MB after 180 min of illumination. Consequently, this work contributes to establishing a simple approach to synthesize highly fluorescent and photocatalytic NCs.

A system for the synthesis of nanoparticles by laser ablation in liquid that is remotely controlled with PC or smartphone.

Nanoparticles find applications in multiple technological and scientific fields, and laser ablation in liquid (LAL) emerged as a versatile method for providing colloidal solutions of nanomaterials with various composition, by a low cost, simple, self-standing, and "green" procedure. However, the use of high energy and high power laser beams is harmful, especially when coupled with flammable or toxic liquids, and in situ operation is required for starting, monitoring the LAL synthesis, and stopping it at the desired point. Here we describe the hardware and software design and the test results of a system for the production of nanoparticles by laser ablation synthesis in liquid solution (LASiS), which is remotely controllable with a personal computer or a smartphone. In this system, laser energy and solution flux are selectable, and the synthesis status can be monitored and managed at any time off site. Only commercially available components and software are employed, making the whole apparatus easily reproducible in any LAL laboratory. The system has proven its reliability in various conditions, including intercontinental remote control experiments. Overall, this apparatus represents a step forward to improve the safety and to more efficiently exploit the time of people working with LASiS, thus contributing to the increasing demand for off-site real time monitoring of experimental equipment in many scientific and industrial laboratories, due to safety and efficiency requirements.

In situ electron microscopy observation of the redox process in plasmonic heterogeneous-photo-sensitive nanoparticles.

Observation of relevant phenomena related with dynamical redox process in a plasmonic heterogeneous-photocatalyst system composed by silver nanoparticles (NPs) around and in contact with amorphous silver chloride NPs are reported by in situ transmission electron microscopy. During this process, nanobubbles are initially produced inside the silver chloride NPs, which immediately begin to move within the amorphous phase. Besides, silver atoms inside the silver chloride NPs start to migrate out the occupied volume leaving a space behind, which is filled by crystalline regions of silver chloride located between the pre-existing silver NPs. During the observation time, fast-nucleation, movement, growth, and fast-dissolution of silver NPs take place. Specific space correlation with silver mass loss (or gain) when a new NP is formed (or dissolved), was detected in different regions during the reaction. This mass loss (or gain) takes place on certain places of pre-existing silver NPs. All these phenomena were observed for a configuration comprising at least two silver NPs separated few nanometers apart by a silver chloride NP.

Clean rhodium nanoparticles prepared by laser ablation in liquid for high performance electrocatalysis of the hydrogen evolution reaction.

Rhodium nanoparticles (NPs) were prepared by a one-step, green and facile procedure consisting in laser ablation of a bulk Rh target immersed in pure water (W-Rh-NPs) or ethanol (E-Rh-NPs). When embedded in mesoporous carbon based inks, both W-Rh-NPs and E-Rh-NPs show excellent activity towards the hydrogen evolution reaction in acidic media, operating close to the thermodynamic potential with 85-97% faradaic yields and low Tafel slopes of 50-54 mV per decade in the low overpotential region (η < 20 mV). A superior activity of W-Rh-NPs with respect to E-Rh-NPs is ascribed to the absence of surface carbon reducible species derived from the synthesis in organic solvent, and thus confirms the importance of the use of water as the preferred medium for laser synthesis of clean nanocrystals in liquid environment. These results provide an important contribution to the impelling need for the preparation of nano-catalysts based on energy critical materials by clean, sustainable and low cost routes.

Optical and Magnetic Properties of Fe Nanoparticles Fabricated by Femtosecond Laser Ablation in Organic and Inorganic Solvents.

Magnetic nanoparticles have attracted much interest due to their broad applications in biomedicine and pollutant remediation. In this work, the optical, magnetic, and structural characteristics of colloids produced by ultrashort pulsed laser ablation of a solid Fe target were studied in four different media: HPLC water, an aqueous solution of trisodium citrate, acetone, and ethanol. Optical extinction spectroscopy revealed an absorption band in the UV region for all, in contrast to the results obtained with nanosecond lasers. Micro-Raman spectroscopy showed that the samples are heterogeneous in their composition, with hematite, maghemite, and magnetite nanoparticles in all four solvents. Similar results were obtained by electron diffraction, which also found α-Fe. Magnetic properties were studied by vibrating-sample magnetometry, and showed nanoparticles in the superparamagnetic state. Under certain experimental conditions, submicrometer-sized iron oxide nanoparticles agglomerate into fractal patterns that show self-similar properties. Self-assembled annular structures on the nanometer scale were also observed and are reported for the first time.

Mesenchymal stromal cells prolong the lifespan in a rat model of amyotrophic lateral sclerosis.

BACKGROUND AIMS: Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disorder characterized by the loss of brain and spinal cord motor neurons (MN). The intraspinal and systemic grafting of mesenchymal stromal cells (MSC) was used to treat symptomatic transgenic rats overexpressing human superoxide dismutase 1 (SOD1) in order to alleviate the disease course and prolong the animals' lifespan. METHODS: At the age of 16 weeks (disease onset) the rats received two grafts of MSC expressing green fluorescent protein (GFP(+) MSC) on the same day, intraspinally (10(5) cells) and intravenously (2 × 10(6) cells). Sham-treated animals were injected with phosphate-buffered saline (PBS). Motor activity, grip strength and body weight were tested, followed by immunohistochemical analysis. RESULTS: The combined grafting of MSC into symptomatic rats had a significant effect on motor activity and grip strength starting 4 weeks after transplantation. The lifespan of animals in the treated group was 190 ± 3.33 days compared with 179 ± 3.6 days in the control group of animals. Treated rats had a larger number of MN at the thoracic and lumbar levels; these MN were of larger size, and the intensity of terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick-end labeling (TUNEL) staining in the somas of apoptotic MN at the thoracic level was much lower than in sham-treated animals. Transplanted GFP(+) MSC survived in the spinal cord until the end stage of the disease and migrated both rostrally and caudally from the injection site. CONCLUSIONS: Intraspinal and intravenous transplantation of MSC has a beneficial and possibly synergistic effect on the lifespan of ALS animals.

Transplantation of predifferentiated adipose-derived stromal cells for the treatment of spinal cord injury.

Adipose-derived stromal cells (ASCs) are an alternative source of stem cells for cell-based therapies of neurological disorders such as spinal cord injury (SCI). In the present study, we predifferentiated ASCs (pASCs) and compared their behavior with naïve ASCs in vitro and after transplantation into rats with a balloon-induced compression lesion. ASCs were predifferentiated into spheres before transplantation, then pASCs or ASCs were injected intraspinally 1 week after SCI. The cells' fate and the rats' functional outcome were assessed using behavioral, histological, and electrophysiological methods. Immunohistological analysis of pASCs in vitro revealed the expression of NCAM, NG2, S100, and p75. Quantitative RT-PCR at different intervals after neural induction showed the up-regulated expression of the glial markers NG2 and p75 and the neural precursor markers NCAM and Nestin. Patch clamp analysis of pASCs revealed three different types of membrane currents; however, none were fast activating Na(+) currents indicating a mature neuronal phenotype. Significant improvement in both the pASC and ASC transplanted groups was observed in the BBB motor test. In vivo, pASCs survived better than ASCs did and interacted closely with the host tissue, wrapping host axons and oligodendrocytes. Some transplanted cells were NG2- or CD31-positive, but no neuronal markers were detected. The predifferentiation of ASCs plays a beneficial role in SCI repair by promoting the protection of denuded axons; however, functional improvements were comparable in both the groups, indicating that repair was induced mainly through paracrine mechanisms.

Co-transplantation of olfactory ensheathing glia and mesenchymal stromal cells does not have synergistic effects after spinal cord injury in the rat.

BACKGROUND AIMS: Olfactory ensheathing glia (OEG) and mesenchymal stromal cells (MSC) are suitable candidates for transplantation therapy of spinal cord injury (SCI). Both facilitate functional improvement after SCI by producing trophic factors and cytokines. In this study, the co-transplantation of both types of cells was studied to clarify their additive and/ or synergistic effects on SCI. METHODS: A balloon-induced compression lesion was used to produce SCI in rats. OEG, MSC or both OEG and MSC (3 x 10(5) cells of each cell type) were implanted by intraspinal injection 1 week after SCI. The effect of transplantation was assessed using behavioral, electrophysiologic and histologic methods. RESULTS: Hindlimb function was examined with Basso, Beattie and Bresnahan (BBB) and Plantar tests. Improvement was found in all three groups of transplanted rats with different time-courses, but there was no significant difference among the groups at the end of the experiment. Motor-evoked potentials after SCI decreased in amplitude from 7 mV to 10 microV. Linear regression analysis showed a modest recovery in amplitude following transplantation, but no change in the control rats. Histologic findings showed that the white and gray matter were significantly spared by transplantation after SCI. CONCLUSIONS: Functional improvement was achieved with transplantation of OEG and/or MSC, but the co-transplantation of OEG and MSC did not show synergistic effects. The poor migration of OEG and MSC might prevent their concerted action. Pre-treatment with a Rho antagonist and a combination of intraspinal and intravenous injection of the cells might be beneficial for SCI therapy.

Evaluación de la actividad antiviral in vitro de cuatro extractos de las especies Caryodendron orinocense y Phyllanthus niruri de la familia Euphorbiaceae contra los virus herpes bovino tipo 1 y herpes simplex tipo 2 / Evaluation of the in vitro antiviral activity of four extracts from the species Caryodendron orinocense and Phyllantus niruri from the Euphorbiaceae family against herpes simplex virus type 1 and bovine herpes virus type 2

Los virus causan enfermedades humanas y animales de gran importancia epidemiológica y económica, para la mayoría de las cuales no existen tratamientos satisfactorios, o con las terapias se generan cepas resistentes. Extractos de plantas pertenecientes a la familia Euphorbiaceae han mostrado actividad contra virus de la familia Herpesviridae. Utilizando la técnica de tinción en placa con cristal violeta, se evaluó la citotoxicidad y la actividad antiviral de los extractos en hexano, en acetato de etilo, en metanol y en agua de las especies Caryodendron orinocense y Phyllanthus niruri contra los virus del Herpes Simplex tipo 2 (HSV-2) y del Herpes Bovino tipo 1 (HVB-1). Además, se realizó un estudio fitoquímico preliminar de las dos especies. En general, los extractos de las especies estudiadas muestran citotoxicidad a concentraciones mayores de 250 μg/mL. El extracto en acetato de etilo de la especie Caryodendron orinocense fue el que registró el mayor factor de reducción viral para HSV-2 y HVB-1, el cual fue de 102 a una concentración de 125 μg/ml y de 104 a 62.5 μg/ml respectivamente. Las especies estudiadas muestran una composición química determinada principalmente por aminoácidos, compuestos fenólicos, taninos y triterpenoides, metabolitos que podrían estar involucrados en su actividad antiviral.

Pathological shear stress stimulates the tyrosine phosphorylation of alpha-actinin associated with the glycoprotein Ib-IX complex.

Shear-induced platelet responses are triggered by VWF binding to the platelet GpIb-IX complex, and there is evidence that this ligand-receptor coupling stimulates transmembranous signaling through the cytoplasmic tail of glycoprotein (Gp) Ib alpha. To investigate the mechanism by which signaling is effected, new molecular interactions involving GpIb-IX that develop in response to pathological shearing stress were examined in intact human platelets. Exposure to shear, but not alpha-thrombin, results in the co-immunoprecipitation of the actin cross-linking protein alpha-actinin with the GpIb-IX complex. Blockers of VWF binding to GpIb alpha or actin polymerization inhibit the association of alpha-actinin with the GpIb-IX complex, but the association of alpha-actinin with the GpIb-IX complex is not affected by inhibiting VWF binding to platelet integrin alpha IIb beta 3 (GpIIb-IIIa). alpha-Actinin becomes tyrosine phosphorylated in response to pathological shear stress, and phosphorylated alpha-actinin associates with GpIb-IX. In resting platelets, class IA heterodimeric phosphatidylinositol 3-kinase (PI 3-K) and protein kinase N (PKN) associate with nonphosphorylated alpha-actinin. Shear stress causes PI 3-K to disassociate from alpha-actinin, while it stimulates PKN binding to alpha-actinin. These results demonstrate that shear-induced VWF binding to GpIb alpha causes enhanced binding of cytoskeletal alpha-actinin to GpIb-IX and suggest that alpha-actinin, perhaps through tyrosine phosphorylation, serves as an adapter for a signaling complex that could regulate VWF-induced platelet aggregation.