Activation of Human Complement System by Dextran-Coated Iron Oxide Nanoparticles Is Not Affected by Dextran/Fe Ratio, Hydroxyl Modifications, and Crosslinking

While having tremendous potential as therapeutic and imaging tools, the clinical use of engineered nanoparticles has been associated with serious safety concerns. Activation of the complement cascade and release of proinflammatory factors C3a and C5a may contribute to infusion-related reactions, whereas opsonization with C3 fragments promotes rapid recognition and clearance of nanomaterials by mononuclear phagocytes. We used dextran-coated superparamagnetic iron oxide nanoparticles (SPIO), which are potent activators of the complement system, to study the role of nanoparticle surface chemistry in inciting complement in human serum. Using complement inhibitors and measuring levels of fluid phase markers (sC5b-9, C5a and Bb), we found that the majority of human complement activation by SPIO is through the alternative pathway (AP). SPIO prepared with high dextran/iron ratio showed some complement activation via calcium-sensitive pathways, but the AP was responsible for the bulk of complement activation and amplification. Activation via the AP required properdin, the positive regulator of the alternative C3bBb convertase. Modification of sugar alcohols of dextran with alkylating, acylating, or crosslinking agents did not overcome complement activation and C3 opsonization. These data demonstrate that human complement activation is independent of dextran modification of SPIO and suggest a crucial role of the AP in immune recognition of nano-assemblies in human serum

Total stimulation gonadotropin dose per oocyte retrieved and fresh embryo quality affect live birth rate after frozen-thawed embryo transfer

Koeputkihedelmöityshoidoissa (IVF) ja munasolujen mikroinjektiohoidoissa (ICSI) elävän lapsen syntymisen todennäköisyyteen vaikuttavia tekijöitä ovat muun muassa kerättyjen munasolujen lukumäärä, saatu gonadotropiinihormonin (FSH) annos sekä luotujen korkealaatuisten alkioiden määrä. Tämän tutkimuksen tarkoituksena oli selvittää, mitkä ominaisuudet alkion tuoresiirtovaiheessa vaikuttavat syntymän todennäköisyyteen jatkossa, jos naiselle tehdään myöhemmin hedelmöityshoidossa pakastetun alkion siirto. Tutkimusaineistona oli yhteinen Suomen lapsettomuusklinikoista kerätty tietokanta, joka sisälsi tiedot yhteensä 9465 pakastusalkionsiirto-tyyppisestä hedelmöityshoidosta vuosilta 2000-2017. Tuorealkionsiirtoja tutkittiin verraten näitä samojen naisten myöhempiin pakastealkionsiirtoihin. Luteaalisen kierron hormonaalista tukea sai naisista 42.9 %, hormonaalisesti indusoitu kuukautiskierto ja ovulaatio oli 40.7 % naisista, ja 16.1 % naisista kuukautiskierto oli täysin spontaani. FSH-annosta/munasolu käytettiin munasarjojen vasteen arviointiin. Tutkimuksessa selvisi, että vertailussa matalin FSH/munasolu -annossuhde <200 IU/munasolu sai aikaan suurimman todennäköisyyden elävän lapsen syntymälle pakastealkionsiirron jälkeen. Korkeampi annossuhde 300-399 IU/munasolu sen sijaan laski merkittävästi tätä todennäköisyyttä. Potilaan hormonaalisesti indusoitu ovulaatiokierto niin ikään assosioitui matalampaan syntymän todennäköisyyteen kuin kierto, jossa oli annettu hormonaalista tukea vain luteaalivaiheessa. Mikäli ainakin yksi korkealaatuinen alkio oli saatavilla hoitojen alussa, oli syntymän todennäköisyys suurempi läpi koko prosessin. Naisen yli 35 vuoden ikä laski syntymän todennäköisyyttä. Hedelmöityshoidoissa FSH/munasolu-annossuhde kuvastaa mahdollisesti munasarjojen toimintaa ja munasolujen laatua. Annossuhde tulisi pyrkiä pitämään mahdollisimman alhaisena, jotta vältytään korkeampiin annoksiin liittyvältä syntymän todennäköisyyden laskulta

Hyperpolarized Long-T1 Silicon Nanoparticles for Magnetic Resonance Imaging

Silicon nanoparticles are experimentally investigated as a potential hyperpolarized, targetable MRI imaging agent. Nuclear T_1 times at room temperature for a variety of Si nanoparticles are found to be remarkably long (10^2 to 10^4 s) - roughly consistent with predictions of a core-shell diffusion model - allowing them to be transported, administered and imaged on practical time scales without significant loss of polarization. We also report surface functionalization of Si nanoparticles, comparable to approaches used in other biologically targeted nanoparticle systems.Comment: supporting material here: http://marcuslab.harvard.edu/Aptekar_hyper1_sup.pd

Phase I Clinical Trial of Systemically Administered TUSC2(FUS1)-Nanoparticles Mediating Functional Gene Transfer in Humans

Background: Tumor suppressor gene TUSC2/FUS1 (TUSC2) is frequently inactivated early in lung cancer development. TUSC2 mediates apoptosis in cancer cells but not normal cells by upregulation of the intrinsic apoptotic pathway. No drug strategies currently exist targeting loss-of–function genetic abnormalities. We report the first in-human systemic gene therapy clinical trial of tumor suppressor gene TUSC2. Methods: Patients with recurrent and/or metastatic lung cancer previously treated with platinum-based chemotherapy were treated with escalating doses of intravenous N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTAP):cholesterol nanoparticles encapsulating a TUSC2 expression plasmid (DOTAP:chol-TUSC2) every 3 weeks. Results: Thirty-one patients were treated at 6 dose levels (range 0.01 to 0.09 milligrams per kilogram). The MTD was determined to be 0.06 mg/kg. Five patients achieved stable disease (2.6–10.8 months, including 2 minor responses). One patient had a metabolic response on positron emission tomography (PET) imaging. RT-PCR analysis detected TUSC2 plasmid expression in 7 of 8 post-treatment tumor specimens but not in pretreatment specimens and peripheral blood lymphocyte controls. Proximity ligation assay, performed on paired biopsies from 3 patients, demonstrated low background TUSC2 protein staining in pretreatment tissues compared with intense (10–25 fold increase) TUSC2 protein staining in posttreatment tissues. RT-PCR gene expression profiling analysis of apoptotic pathway genes in two patients with high posttreatmen

Antimicrobial Nanoplexes meet Model Bacterial Membranes: the key role of Cardiolipin

Antimicrobial resistance to traditional antibiotics is a crucial challenge of medical research. Oligonucleotide therapeutics, such as antisense or Transcription Factor Decoys (TFDs), have the potential to circumvent current resistance mechanisms by acting on novel targets. However, their full translation into clinical application requires efficient delivery strategies and fundamental comprehension of their interaction with target bacterial cells. To address these points, we employed a novel cationic bolaamphiphile that binds TFDs with high affinity to form self-assembled complexes (nanoplexes). Confocal microscopy revealed that nanoplexes efficiently transfect bacterial cells, consistently with biological efficacy on animal models. To understand the factors affecting the delivery process, liposomes with varying compositions, taken as model synthetic bilayers, were challenged with nanoplexes and investigated with Scattering and Fluorescence techniques. Thanks to the combination of results on bacteria and synthetic membrane models we demonstrate for the first time that the prokaryotic-enriched anionic lipid Cardiolipin (CL) plays a key-role in the TFDs delivery to bacteria. Moreover, we can hypothesize an overall TFD delivery mechanism, where bacterial membrane reorganization with permeability increase and release of the TFD from the nanoplexes are the main factors. These results will be of great benefit to boost the development of oligonucleotides-based antimicrobials of superior efficacy

Voice disorders and mental health in teachers: a cross-sectional nationwide study

International audienceBACKGROUND:Teachers, as professional voice users, are at particular risk of voice disorders. Among contributing factors, stress and psychological tension could play a role but epidemiological data on this problem are scarce. The aim of this study was to evaluate prevalence and cofactors of voice disorders among teachers in the French National Education system, with particular attention paid to the association between voice complaint and psychological status.METHODS:The source data come from an epidemiological postal survey on physical and mental health conducted in a sample of 20,099 adults (in activity or retired) selected at random from the health plan records of the national education system. Overall response rate was 53%. Of the 10,288 respondents, 3,940 were teachers in activity currently giving classes to students. In the sample of those with complete data (n = 3,646), variables associated with voice disorders were investigated using logistic regression models. Studied variables referred to demographic characteristics, socio-professional environment, psychological distress, mental health disorders (DSM-IV), and sick leave.RESULTS:One in two female teachers reported voice disorders (50.0%) compared to one in four males (26.0%). Those who reported voice disorders presented higher level of psychological distress. Sex- and age-adjusted odds ratios [95% confidence interval] were respectively 1.8 [1.5-2.2] for major depressive episode, 1.7 [1.3-2.2] for general anxiety disorder, and 1.6 [1.2-2.2] for phobia. A significant association between voice disorders and sick leave was also demonstrated (1.5 [1.3-1.7]).CONCLUSION:Voice disorders were frequent among French teachers. Associations with psychiatric disorders suggest that a situation may exist which is more complex than simple mechanical failure. Further longitudinal research is needed to clarify the comorbidity between voice and psychological disorders

Characterization of monoolein-based lipoplexes using fluorescence spectroscopy

Lipoplexes are commonly used as delivery systems in vitro and in vivo, the role of a neutral lipid as helper being of extreme importance in these systems. Cationic liposomes composed of dioctadecyldimethylammonium bromide (DODAB) with monoolein (MO) as a helper, at different molar ratios (1:2; 1:1 and 1:0.5) were prepared, and subsequently titrated to DNA. The structural and physicochemical properties of the lipid/DNA complexes were assessed by Ethidium Bromide (EtBr) exclusion, 90º Static Light Scattering (90º SLS) assays and Fluorescence Resonance Energy Transfer (FRET). In EtBr exclusion assays, the steady-state fluorescence spectra of EtBr were decomposed into the sum of two lognormal emissions, emanating from two different environments – H2O and DNA, and the effect of charge ratio (+/-) was observed. 90º SLS assays gave an important contribution, detecting size variations in systems with different MO fractions on the lipoplexes. In FRET assays, 2-(3-(diphenylhexatrienyl)propanoyl)-1-hexadecanoyl-sn-glycero-3-phosphocholine (DPH-HPC) was used as donor and EtBr as acceptor. The DNA component previously calculated by EtBr exclusion, was used to determine the energy transfer efficiency, as an indirect measurement of the lipoplexes structural and physicochemical properties. Our results demonstrate that the inclusion of monoolein in the cationic liposomes formulation significantly modifies the rate of DNA complexation, being DODAB:MO (1:1) the system with higher DNA condensation efficiency.Fundação para a Ciência e a Tecnologia (FCT

New FTY720-docetaxel nanoparticle therapy overcomes FTY720-induced lymphopenia and inhibits metastatic breast tumour growth

Purpose: Combining molecular therapies with chemotherapy may offer an improved clinical outcome for chemoresistant tumours. Sphingosine-1-phosphate (S1P) receptor antagonist and sphingosine kinase 1 (SK1) inhibitor FTY720 (FTY) has promising anticancer properties, however, it causes systemic lymphopenia which impairs its use in cancer patients. In this study, we developed a nanoparticle (NP) combining docetaxel (DTX) and FTY for enhanced anticancer effect, targeted tumour delivery and reduced systemic toxicity. Methods: Docetaxel, FTY and glucosamine were covalently conjugated to poly(lactic-co-glycolic acid) (PLGA). NPs were characterised by dynamic light scattering and electron microscopy. The cellular uptake, cytotoxicity and in vivo antitumor efficacy of CNPs were evaluated. Results: We show for the first time that in triple negative breast cancer cells FTY provides chemosensitisation to DTX, allowing a four-fold reduction in the effective dose. We have encapsulated both drugs in PLGA complex NPs (CNPs), with narrow size distribution of ~ 100 nm and excellent cancer cell uptake providing sequential, sustained release of FTY and DTX. In triple negative breast cancer cells and mouse breast cancer models, CNPs had similar efficacy to systemic free therapies, but allowed an effective drug dose reduction. Application of CNPs has significantly reversed chemotherapy side effects such as weight loss, liver toxicity and, most notably, lymphopenia. Conclusions: We show for the first time the DTX chemosensitising effects of FTY in triple negative breast cancer. We further demonstrate that encapsulation of free drugs in CNPs can improve targeting, provide low off-target toxicity and most importantly reduce FTY-induced lymphopenia, offering potential therapeutic use of FTY in clinical cancer treatment