24 research outputs found
Broadband stimulated Raman imaging based on multi-channel lock-in detection for spectral histopathology
Spontaneous Raman microscopy reveals the chemical composition of a sample in a label-free and non-invasive fashion by directly measuring the vibrational spectra of molecules. However, its extremely low cross section prevents its application to fast imaging. Stimulated Raman scattering (SRS) amplifies the signal by several orders of magnitude thanks to the coherent nature of the nonlinear process, thus unlocking high-speed microscopy applications that provide analytical information to elucidate biochemical mechanisms with subcellular resolution. Nevertheless, in its standard implementation, narrowband SRS provides images at only one frequency at a time, which is not sufficient to distinguish constituents with overlapping Raman bands. Here, we report a broadband SRS microscope equipped with a home-built multichannel lock-in amplifier simultaneously measuring the SRS signal at 32 frequencies with integration time down to 44 ÎĽs, allowing for detailed, high spatial resolution mapping of spectrally congested samples. We demonstrate the capability of our microscope to differentiate the chemical constituents of heterogeneous samples by measuring the relative concentrations of different fatty acids in cultured hepatocytes at the single lipid droplet level and by differentiating tumor from peritumoral tissue in a preclinical mouse model of fibrosarcoma
Zwitterionic nanofibers of super-glue for transparent and biocompatible multi-purpose coatings
Here we show that macrozwitterions of poly(ethyl 2-cyanoacrylate), commonly called Super Glue, can easily assemble into long and well defined fibers by electrospinning. The resulting fibrous networks are thermally treated on glass in order to create transparent coatings whose superficial
morphology recalls the organization of the initial electrospun mats. These textured coatings are characterized by low liquid adhesion and anti-staining performance. Furthermore, the low friction coefficient and excellent scratch resistance make them attractive as solid lubricants. The inherent
texture of the coatings positively affects their biocompatibility. In fact, they are able to promote the proliferation and differentiation of myoblast stem cells. Optically-transparent and biocompatible
coatings that simultaneously possess characteristics of low water contact angle hysteresis, low friction and mechanical robustness can find application in a wide range of technological sectors, from
the construction and automotive industries to electronic and biomedical devices
Neural stem cell transplantation in patients with progressive multiple sclerosis: an open-label, phase 1 study
Innovative pro-regenerative treatment strategies for progressive multiple sclerosis (PMS), combining neuroprotection and immunomodulation, represent an unmet need. Neural precursor cells (NPCs) transplanted in animal models of multiple sclerosis have shown preclinical efficacy by promoting neuroprotection and remyelination by releasing molecules sustaining trophic support and neural plasticity. Here we present the results of STEMS, a prospective, therapeutic exploratory, non-randomized, open-label, single-dose-finding phase 1 clinical trial (NCT03269071, EudraCT 2016-002020-86), performed at San Raffaele Hospital in Milan, Italy, evaluating the feasibility, safety and tolerability of intrathecally transplanted human fetal NPCs (hfNPCs) in 12 patients with PMS (with evidence of disease progression, Expanded Disability Status Scale >= 6.5, age 18-55 years, disease duration 2-20 years, without any alternative approved therapy). The safety primary outcome was reached, with no severe adverse reactions related to hfNPCs at 2-year follow-up, clearly demonstrating that hfNPC therapy in PMS is feasible, safe and tolerable. Exploratory secondary analyses showed a lower rate of brain atrophy in patients receiving the highest dosage of hfNPCs and increased cerebrospinal fluid levels of anti-inflammatory and neuroprotective molecules. Although preliminary, these results support the rationale and value of future clinical studies with the highest dose of hfNPCs in a larger cohort of patients
Stable population structure in Europe since the Iron Age, despite high mobility
Ancient DNA research in the past decade has revealed that European population structure changed dramatically in the prehistoric period (14,000–3000 years before present, YBP), reflecting the widespread introduction of Neolithic farmer and Bronze Age Steppe ancestries. However, little is known about how population structure changed from the historical period onward (3000 YBP - present). To address this, we collected whole genomes from 204 individuals from Europe and the Mediterranean, many of which are the first historical period genomes from their region (e.g. Armenia and France). We found that most regions show remarkable inter-individual heterogeneity. At least 7% of historical individuals carry ancestry uncommon in the region where they were sampled, some indicating cross-Mediterranean contacts. Despite this high level of mobility, overall population structure across western Eurasia is relatively stable through the historical period up to the present, mirroring geography. We show that, under standard population genetics models with local panmixia, the observed level of dispersal would lead to a collapse of population structure. Persistent population structure thus suggests a lower effective migration rate than indicated by the observed dispersal. We hypothesize that this phenomenon can be explained by extensive transient dispersal arising from drastically improved transportation networks and the Roman Empire’s mobilization of people for trade, labor, and military. This work highlights the utility of ancient DNA in elucidating finer scale human population dynamics in recent history
Development of a multi channel acquisition scheme for a broadband Stimulated Raman Scattering microscopy system
Coherent Raman scattering (CRS) microscopy has emerged as a high-speed, high sensitivity vibrational imaging platform. Stimulated Raman scattering (SRS) microscopes, in particular, with their subcellular resolution, combined with the capability to collect precise vibrational signatures, are allowing to perform quantitative chemical imaging even in live cells. Being intrinsically label-free, vibrational spectroscopic microscopes allow the measurement of biological samples without perturbing the function of molecules. SRS has the advantage to be background free and thus directly comparable with spontaneous Raman scattering, giving access to the wide catalog of spectra already existing in the literature.
The aim of this thesis work is to characterize and improve the signal to noise ratio measurement of an SRS microscope which exploits spectral shaping by a narrowband and rapidly tunable Acousto Optical Tunable Filter. A multichannel approach, performing Hadamard reconstruction method, has been implemented for spectra acquisition, and its performance evaluated.
The configuration developed allows performing a spectral scanning from the Raman fingerprint region to the CH-stretch region without any change of the optical setup therefore ensuring high acquisition speed. Application of the SRS microscope to broadband hyperspectral imaging of biological samples is demonstrated
Full season efficacy of moxidectin microsphere sustained release formulation for the prevention of heartworm (Dirofilaria immitis) infection in dogs
The authors report the efficacy of an injectable, moxidectin sustained release (SR) formulation for the prevention of canine heartworm infection in endemic areas in northern and central Italy. Three field trials were carried out on a total of 324 dogs. Two hundred forty-three dogs were treated with moxidectin SR 6 months apart and 81 dogs (positive controls) with moxidectin tablets given monthly for 5 consecutive months during the risk season each year throughout the study. Results of testing for microfilariae and circulating adult female antigens were negative for all the experimentally treated dogs at the 6, 7, 11 and 19 months after the last injection. No adverse reactions to moxidectin SR were observed but a moderate pain at palpation and swelling (5-6 cm) at the injection site after the first treatment. In the study areas, prevalence of Dirofilaria immitis infection calculated by testing dogs which had no preventive treatment in the previous transmission season ranged from 33 to 63%. This study confirms the efficacy and safety of injectable, moxidectin SR formulation in the prevention of heartworm infection in dogs and demonstrates that the prophylactic efficacy lasts for the full season and strongly suggests that the product gives 1-year protection
P(VDF-TrFE)/BaTiO3 Nanoparticle Composite Films Mediate Piezoelectric Stimulation and Promote Differentiation of SH-SY5Y Neuroblastoma Cells
Poly(vinylidene fluoride-trifluoroethylene, P(VDF-TrFE)) and P(VDF-TrFE)/barium titanate nanoparticle (BTNP) films are prepared and tested as substrates for neuronal stimulation through direct piezoelectric effect. Films are characterized in terms of surface, mechanical, and piezoelectric features before in vitro testing on SH-SY5Y cells. In particular, BTNPs significantly improve piezoelectric properties of the films (4.5-fold increased d31 ). Both kinds of films support good SH-SY5Y viability and differentiation. Ultrasound (US) stimulation is proven to elicit Ca(2+) transients and to enhance differentiation in cells grown on the piezoelectric substrates. For the first time in the literature, this study demonstrates the suitability of polymer/ceramic composite films and US for neuronal stimulation through direct piezoelectric effect
Dinamiche relazionali familiari e obesit\ue0 in infanzia e adolescenza: i modelli family-oriented
Dinamiche relazionali familiari e obesit\ue0 in infanzia e adolescenza: i modelli family-oriente
A New Phase of Research on the Protohistoric Graves of Swat Valley, Khyber Pukhtunkhwa, Pakistan
Bio/non-bio interfaces: a straightforward method for obtaining long term PDMS/muscle cell biohybrid constructs
Stable surface modifications of polydimethylsiloxane (PDMS) are of crucial importance for the exploitation of the versatile physical properties of silicone in many biological applications. Surface hydrophobic recovery in fact poses severe time limitations to the observation of biological events and, in particular, to cell culturing. A novel method of stable modification of PDMS surface chemistry was therefore elaborated, relying on the use of genipin as a natural low-toxicity cross-linker, and involving free amine moieties. Its effectiveness to long-term cultures was studied by preparation of thin PDMS films with different stiffness. After assessment of surface chemistry and substrate stiffness, H9c2 muscle cells were cultured on the modified films, and differentiating myoblasts were observed for a period of four weeks since differentiation induction. A lower PDMS stiffness increased myotube width and supported a higher actin and myosin colocalization within myotubes, suggesting the achievement of myotube functional maturity. These results provide evidence of the effectiveness of the proposed procedures to PDMS surface chemistry modification. Furthermore, modified PDMS membranes prove to be suitable to several long-term studies of cell behaviour in vitro, including muscle cell contractility investigations