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Surface-enhanced Raman scattering (SERS) is of growing interest for a wide range of applications, especially for biomedical analysis, thanks to its sensitivity, specificity, and multiplexing capabilities. A crucial role for successful applications of SERS is played by the development of reproducible, efficient, and facile procedures for the fabrication of metal nanostructures (SERS substrates). Even more challenging is to extend the fabrication techniques of plasmonic nano-textures to atomic force microscope (AFM) probes to carry out tip-enhanced Raman spectroscopy (TERS) experiments, in which spatial resolution below the diffraction limit is added to the peculiarities of SERS. In this short review, we describe recent studies performed by our group during the last ten years in which novel nanofabrication techniques have been successfully applied to SERS and TERS experiments for studying bio-systems and molecular species of environmental interest. [ABSTRACT FROM AUTHOR]

Nanoparticles (NPs) coated with hyaluronic acid (HA) seem to be increasingly promising for targeted therapy due to HA chemical versatility, which allows them to bind drugs of different natures, and their affinity with the transmembrane receptor CD-44, overexpressed in tumor cells. However, an essential aspect for clinical use of NPs is formulation stability over time. For these reasons, analytical techniques capable of characterizing their physico-chemical properties are needed. In this work, poly(lactide-co-glycolide) (PLGA) NPs with an average diameter of 100–150 nm, coated with a few 10 s of nm of HA, were synthesized. For stability characterization, two complementary investigative techniques were used: Dynamic Light Scattering (DLS) and Surface-Enhanced Raman Scattering (SERS) spectroscopy. The first technique provided information on size, polidispersity index, and zeta-potential, and the second provided a deeper insight on the NP surface chemicals, allowing distinguishing of HA-coated NPs from uncoated ones. Furthermore, in order to estimate formulation stability over time, NPs were measured and monitored for two weeks. SERS results showed a progressive decrease in the signal associated with HA, which, however, is not detectable by the DLS measurements. [ABSTRACT FROM AUTHOR]

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous pollutants that are typically released into the environment during the incomplete combustion of fossil fuels. Due to their relevant carcinogenicity, mutagenicity, and teratogenicity, it is urgent to develop sensitive and cost-effective strategies for monitoring them, especially in aqueous environments. Surface-enhanced Raman spectroscopy (SERS) can potentially be used as a reliable approach for this purpose, as it constitutes a valid alternative to traditional techniques, such as liquid and gas chromatography. Nevertheless, the development of an SERS-based platform for detection PAHs has so far been hindered by the poor adsorption of PAHs onto silver- and gold-based SERS-active substrates. To overcome this limitation, several research efforts have been directed towards the development of functionalized SERS substrates for the improvement of PAH adsorption. However, these strategies suffer from the interference that functionalizing molecules can produce in SERS detection. Herein, we demonstrate the feasibility of label-free detection of pyrene by using a highly porous 3D-SERS substrate produced by an inductively coupled plasma (ICP). Thanks to the coral-like nanopattern exhibited by our substrate, clear signals ascribable to pyrene molecules can be observed with a limit of detection of 23 nM. The observed performance can be attributed to the nanoporous character of our substrate, which combines a high density of hotspots and a certain capability of trapping molecules and favoring their adhesion to the Ag nanopattern. The obtained results demonstrate the potential of our substrates as a large-area, label-free SERS-based platform for chemical sensing and environmental control applications. [ABSTRACT FROM AUTHOR]

The fabrication of plasmonic nanostructures with a reliable, low cost and easy approach has become a crucial and urgent challenge in many fields, including surface-enhanced Raman spectroscopy (SERS) based applications. In this frame, nanoporous metal films are quite attractive, due to their intrinsic large surface area and high density of metal nanogaps, acting as hot-spots for Raman signal enhancement. In this paper, we report a detailed study on the fabrication of nanoporous silver-based SERS substrates, obtained by the application of two successive treatments with an Inductively Coupled Plasma (ICP) system, using synthetic air and Ar as feeding gases. The obtained substrates exhibit a quite broad plasmonic response, covering the Vis–NIR range, and an enhancement factor reaching 6.5 × 10 7 , estimated by using 4-mercaptobenzoic acid (4-MBA) as probe molecule at 532 nm. Moreover, the substrates exhibit a quite good spatial reproducibility on a centimeter scale, which assures a good signal stability for analytical measurements. Globally, the developed protocol is easy and cost effective, potentially usable also for mass production thanks to the remarkable inter-batches reproducibility. As such, it holds promise for its use in SERS-based sensing platforms for sensitive detection of targets molecules. [ABSTRACT FROM AUTHOR]

The development of sensitive methods for the detection of endotoxin molecules, such as lipopolysaccharides (LPS), is essential for food safety and health control. Conventional analytical methods used for LPS detection are based on the pyrogen test, plating and culture-based methods, and the limulus amoebocyte lysate method (LAL). Alternatively, the development of reliable biosensors for LPS detection would be highly desirable to solve some critical issues, such as high cost and a long turnaround time. In this work, we present a label-free Surface-Enhanced Raman Spectroscopy (SERS)-based method for LPS detection in its free form. The proposed method combines the benefits of plasmonic enhancement with the selectivity provided by a specific anti-lipid A antibody (Ab). A high-enhancing nanostructured silver substrate was coated with Ab. The presence of LPS was quantitatively monitored by analyzing the changes in the Ab spectra obtained in the absence and presence of LPS. A limit of detection (LOD) and quantification (LOQ) of 12 ng/mL and 41 ng/mL were estimated, respectively. Importantly, the proposed technology could be easily expanded for the determination of other biological macromolecules. [ABSTRACT FROM AUTHOR]

Suitable metal nanostructures may induce surface-enhanced Raman scattering (SERS) enhancement factors (EFs) large-enough to reach single-molecule sensitivity. However, the gap hot-spot EF probability density function (PDF) has the character of a long-tail distribution, which dramatically mines the reproducibility of SERS experiments. Herein, we carry out electrodynamic calculations based on a 3D finite element method of two plasmonic nanostructures, combined with Monte Carlo simulations of the EF statistics under different external conditions. We compare the PDF produced by a homodimer of nanoparticles with that provided by a self-similar trimer. We show that the PDF is sensitive to the spatial distribution of near-field enhancement specifically supported by the nanostructure geometry. Breaking the symmetry of the plasmonic system is responsible for inducing particular modulations of the PDF tail resembling a multiple Poisson distribution. We also study the influence that molecular diffusion towards the hottest hot-spot, or selective hot-spot targeting, might have on the EF PDF. Our results quantitatively assess the possibility of designing the response of a SERS substrate so as to contain the intrinsic EF PDF variance and significantly improving, in principle, the reproducibility of SERS experiments. [ABSTRACT FROM AUTHOR]

Tear fluid is a heterogeneous solution containing mainly proteins, lipids, mucins and electrolytes, which regulates the physiology of the human eye. The complex composition of tears can be altered in the presence of eye inflammations. The use of contact lenses is one of the most frequent causes of inflammatory responses of the eye, with the related discomfort often causing the wearer to give up using them. In this paper, we exploit the potentiality of Raman Spectroscopy to analyse the biochemical changes in tear fluid in a contact lens wearer. In particular, we analysed the tear fluid collected from a volunteer as a function of the wearing time for two types of monthly contact lenses (Hydrogel and Si-Hydrogel). Our experimental results show an alteration of the relative concentrations of proteins and lipids in both of the analysed cases. More importantly, our results highlight the diagnostic sensitivity of Raman analysis to select the proper contact lens type for each wearer and optimise the lens wearing conditions. [ABSTRACT FROM AUTHOR]

The pursuit of environmentally friendly solvents has become an essential research topic in sustainable chemistry and nanomaterial science. With the need to substitute toxic solvents in nanofabrication processes becoming more pressing, the search for alternative solvents has taken on a crucial role in this field. Additionally, the use of toxic, non-economical organic solvents, such as N-methyl-2 pyrrolidone and dimethylformamide, is not suitable for all biomedical applications, even though these solvents are often considered as the best exfoliating agents for nanomaterial fabrication. In this context, the success of producing two-dimensional transition metal dichalcogenides (2D TMDs), such as MoS2 and WS2, with excellent captivating properties is due to the ease of synthesis based on environment-friendly, benign methods with fewer toxic chemicals involved. Herein, we report for the first time on the use of cyrene as an exfoliating agent to fabricate monolayer and few-layered 2D TMDs with a versatile, less time-consuming liquid-phase exfoliation technique. This bio-derived, aprotic, green and eco-friendly solvent produced a stable, surfactant-free, concentrated 2D TMD dispersion with very interesting features, as characterized by UV–visible and Raman spectroscopies. The surface charge and morphology of the fabricated nanoflakes were analyzed using ς-potential and scanning electron microscopy. The study demonstrates that cyrene is a promising green solvent for the exfoliation of 2D TMD nanosheets with potential advantages over traditional organic solvents. The ability to produce smaller-sized—especially in the case of WS2 as compared to MoS2—and mono/few-layered nanostructures with higher negative surface charge values makes cyrene a promising candidate for various biomedical and electronic applications. Overall, the study contributes to the development of sustainable and environmentally friendly methods for the production of 2D nanomaterials for various applications. [ABSTRACT FROM AUTHOR]

We investigate a quasi-2D suspension of Brownian particles in an optical speckle field produced by holographic manipulation of a laser wavefront. This system was developed to study, in a systematic and controllable way, a distinctive instance of diffusion, called Fickian yet Non Gaussian diffusion (FnGD), observed, during the last decade, for colloidal particles in a variety of complex and biological fluids. Our setup generates an optical speckle field that behaves like a disordered set of optical traps. First, we describe the experimental setup and the dynamics of the particles, focusing on mean square displacements, displacement distributions and kurtosis. Then, we present Brownian Dynamics simulations of point-like particles in a complex energy landscape, mimicking that generated by the optical speckle field. We show that our simulations can capture the salient features of the experimental results, including the emergence of FnGD, also covering times longer than the ones so far achieved in experiments. Some deviations are observed at long time only, with the Gaussian restoring being slower in simulations than in experiments. Overall, the introduced numerical model might be exploited to guide the design of upcoming experiments targeted, for example, to fully monitor the recovery of Gaussianity. [ABSTRACT FROM AUTHOR]

Abstract: In this work we report on the simultaneous measurement of the hydrodynamic coefficient and the electric charge of single Bacillus subtilis spores. The latter has great importance in protein binding to spores and in the adhesion of spores onto surfaces. The charge and the hydrodynamic coefficient were measured by an accurate procedure based on the analysis of the motion of single spores confined by an optical trap. The technique has been validated using charged spherical polystyrene beads. The excellent agreement of our results with the expected values demonstrates the quality of our procedure. We measured the charge of spores of B. subtilis purified from a wild type strain and from two isogenic mutants characterized by an altered spore surface. Our technique is able to discriminate the three spore types used, by their charge and by their hydrodynamic coefficient which is related to the hydrophobic properties of the spore surface. [Copyright &y& Elsevier]

We describe a general way how to calculate optical forces acting on Rayleigh particles or colloids placed into general interference field. In this paper, we focus on a configuration with three interfering beams laying in one plane and we present a comprehensive analysis of trap properties and particle behaviour. We found that this arrangement can be used for simultaneous 2D/3D confinement or sorting of particles having refractive index higher or lower comparing to the surrounding immersion medium. [Copyright &y& Elsevier]

Micrometer-sized rod-like non-birefringent transparent particles trapped by an astigmatic laser beam having elliptical intensity profile spontaneously rotate until they become aligned with the major axis of the beam cross-section. The observed rotation confirms a simple picture based on orbital angular momentum transfer from the light beam to the trapped particle. A model which regards the particle as a thin cylindrical lens with effective focal lengths f1 and f2 has allowed to calculate the applied optical torque and, hence, the time law governing the particle alignment. The time evolution of the particle alignment is studied for different rod-like particles by analysing the sequence of frames recorded by a CCD camera. The time evolution of the angle α formed between the main axes of the body and the laser shape results in good agreement with the theory for bodies fully immersed in the spot size of the laser. Discrepancies observed for rod-like particles longer than the minor elliptical axis of the beam cross-section, were removed by developing a more general model taking into account the partial overlapping between the body and the laser profile. [Copyright &y& Elsevier]

While the phase behavior of aqueous surfactant solutions is usually described in term of the equilibrium microstructures of lyotropic liquid crystals, the transformations which take place when a phase turns into another one, either by changing the concentration or the temperature, are still to be elucidated. A simultaneous determination of concentration and microstructure is at order to elucidate the phase behavior under changing conditions, such as in a dissolution experiment. Confocal Raman micro-spectroscopy and time-lapse polarized light microscopy are combined to study the phase transitions taking place in the dissolution of a common anionic surfactant (sodium laurylethersulfate) in water. By comparing Raman concentration profiles and polarized light images, it is found that the aqueous solution, with initial surfactant concentration of 72 wt%, undergoes a sequence of complex microstructural transformations including distortion of the initial lamellar phase, formation of an intermediate striated texture, which can be considered as a precursor of a cubic phase, and a heterogeneous hexagonal phase going through a transition region before turning into a micellar phase. The effects of the sodium counter-ion and of water confinement are also investigated by analyzing the OH-stretching bands. [ABSTRACT FROM AUTHOR]

Benzodiazepines, psychotropic drugs, are among the most frequently found pharmaceuticals in aquatic matrices. An increasing number of studies are reporting their harmful effects on adults' behaviour and physiology, while little information is available regarding developing organisms exposed since early stages. Improper activation of GABA receptors during embryonic development is likely to induce relevant consequences on the morphogenesis and, at later stages, on behaviour. This study investigated the negative effects of three increasing concentrations of delorazepam on Xenopus laevis retinal and skeletal muscle morphogenesis. Morphological and ultrastructural investigations were correlated with gene expression, while Raman spectroscopy highlighted the main biochemical components affected. Conventional phototactic response and orientation in the magnetic field were assessed as indicators of proper interaction between sensory organs and the nervous system. Results confirm the profound impact of delorazepam on development and return an alarming picture of the amphibians' survival potentialities in a benzodiazepine-contaminated environment. [Display omitted] • Delorazepam belongs to benzodiazepines, emerging water pollutant class. • Its impact was evaluated on Xenopus laevis embryos. • Delorazepam induced retinal dysmorphogenesis, with visual performance impairment. • Skeletal muscle was damaged, displaying disorganization of fibers and sarcomeres. • Raman spectroscopy revealed significant changes in biochemical components. [ABSTRACT FROM AUTHOR]

A popular picture of the intermittent dynamics of glassy materials depicts each particle as localized in a temporary cage formed by its neighbors, and seldom jumping to another cage. Drawing on experiments on hard sphere colloidal glasses and molecular dynamic simulations of supercooled liquids, here we show how this simplified scenario leads to quantitative relations between single particle motion and macroscopic properties, within a Continuous Time Random Walk (CTRW) framework. For example, the diffusivity and the relaxation time at different length scales can be directly predicted from the jump properties. In the accessible range of temperatures and volume fractions, these predictions work well, although the CTRW formalism neglects the spatially heterogeneous dynamics of glassy systems. In this respect, we show how these dynamic heterogeneities can be highlighted from a study of the correlations between jumps. Finally, the intermittent motion is exploited to infer correlations between local structure and dynamics. [ABSTRACT FROM AUTHOR]

Although oocyte cryopreservation has great potentials in the field of reproductive technologies, it still is an open challenge in the majority of domestic animals and little is known on the biochemical transformation induced by this process in the different cellular compartments. Raman micro-spectroscopy allows the non-invasive evaluation of the molecular composition of cells, based on the inelastic scattering of laser photons by vibrating molecules. The aim of this work was to assess the biochemical modifications of both the zona pellucida and cytoplasm of vitrified/warmed in vitro matured bovine oocytes at different post-warming times. By taking advantage of Principal Component Analysis, we were able to shed light on the biochemical transformation induced by the cryogenic treatment, also pointing out the specific role of cryoprotective agents (CPs). Our results suggest that vitrification induces a transformation of the protein secondary structure from the α-helices to the β-sheet form, while lipids tend to assume a more packed configuration in the zona pellucida. Both modifications result in a mechanical hardening of this cellular compartment, which could account for the reduced fertility rates of vitrified oocytes. Furthermore, biochemical modifications were observed at the cytoplasmic level in the protein secondary structure, with α-helices loss, suggesting cold protein denaturation. In addition, a decrease of lipid unsaturation was found in vitrified oocytes, suggesting oxidative damages. Interestingly, most modifications were not observed in oocytes exposed to CPs, suggesting that they do not severely affect the biochemical architecture of the oocyte. Nevertheless, in oocytes exposed to CPs decreased developmental competence and increased reactive oxygen species production were observed compared to the control. A more severe reduction of cleavage and blastocyst rates after in vitro fertilization was obtained from vitrified oocytes. Our experimental outcomes also suggest a certain degree of reversibility of the induced transformations, which renders vitrified oocytes more similar to untreated cells after 2 h warming. [ABSTRACT FROM AUTHOR]

Background G-quadruplex DNA is involved in many physiological and pathological processes. Both clinical and experimental studies on DNA G-quadruplexes are slowed down by their enzymatic instability. In this frame, more stable chemically modified analogs are needed. Methods The bis-end-linked-(gggt) 2 PNA molecule (BEL-PNA) was synthesized using in solution and solid phase synthetic approaches. Quadruplex formation was assessed by circular dichroism (CD) and surface enhanced Raman scattering (SERS). Results An unprecedented bimolecular PNA homo quadruplex is here reported. To achieve this goal, we developed a bifunctional linker that once functionalized with gggt PNA strands and annealed in K + buffer allowed the obtainment of a PNA homo quadruplex. The identification of the strong SERS band at ~ 1481 cm − 1 , attributable to vibrations involving the quadruplex diagnostic Hoogsteen type hydrogen bonds, confirmed the formation of the PNA homo quadruplex. Conclusions By tethering two G-rich PNA strands to the two ends of a suitable bifunctional linker it is possible to obtain bimolecular PNA homo quadruplexes after annealing in K + -containing buffers. The formation of such CD-unfriendly complexes can be monitored, even at low concentrations, by using the SERS technique. General significance Given the importance of DNA G-quadruplexes in medicine and nanotechnology, the obtainment of G-quadruplex analogs provided with enhanced enzymatic stability, and their monitoring by highly sensitive label-free techniques are of the highest importance. This article is part of a Special Issue entitled "G-quadruplex" Guest Editor: Dr. Concetta Giancola and Dr. Daniela Montesarchio. [ABSTRACT FROM AUTHOR]

Background About twenty variants of apolipoprotein A-I (ApoA-I) are associated to hereditary systemic amyloidoses. Although the molecular bases of this disease are still largely unknown, it has been hypothesized that ApoA-I proteolysis is a key event in pathogenesis, since it triggers the release of an N-terminal fragment (80–100 residue long) that misfolds to form amyloid deposits in peripheral organs and tissues. It is also known that cell membrane lipids play a key role in the fibrillogenic pathway. In the case of ApoA-I related amyloidosis caused by L174S mutation, the 93-residue N-terminal fragment of ApoA-I ([1-93]ApoA-I) was found to be the major constituent of ex vivo fibrils. Methods With the main goal to investigate the interaction of either [1-93]ApoA-I and ApoA-I with biomimetic membranes, we set-up an experimental system based on the Raman Tweezers methodology. We tested GUVs composed by two types of zwitterionic lipids with a different fluidity degree, i.e. dioleoylphosphatidylcholine (DOPC) and dipalmitoylphosphatidylcholine (DPPC). Results We found that [1-93]ApoA-I induces conformational disorder in an ordered lipid bilayer. When interacting with fluid phases, instead, the fragment was found to be able to penetrate the membrane bilayer inducing an alignment of lipid chains. Conclusions The interaction features of [1-93]ApoA-I with biomimetic membranes strongly depend on the lipid phase. Full-length ApoA-I was found to have similar effects, even if significantly less pronounced. General significance Our observations shed light on still largely unknown molecular bases of ApoA-I fibrillogenic domain interaction with membranes. [ABSTRACT FROM AUTHOR]

Disordered hyperuniform (DH) media have been recognized as a new state of disordered matter that broadens our vision of material engineering. Here, long-range correlated disordered two-dimensional patterns are fabricated by self-assembling of spherical diblock-copolymer (BCP) micelles. Control of the self-assembling parameters leads to the formation of DH patterns of micelles that can host nanoscale material inclusions, therefore providing an effective strategy for fabricating multimaterial DH structures at molecular scale. Centroidal patterns are accurately determined by virtue of BCP micelles loaded with metal nanoparticles. Our analysis reveals the signature of nearly ideal DH BCP assemblies in the local density fluctuation and a dominant linear scaling in the local number fluctuation. [ABSTRACT FROM AUTHOR]

Bacterial spores are surrounded by the coat, a multilayered shell that contributes in protecting the genome during stress conditions. In Bacillus subtilis, the model organism for spore formers, the coat is composed by about seventy different proteins, organized into four layers by the action of several regulatory proteins. A major component of this regulatory network, CotE, is needed to assemble the outer coat and develop spores fully resistant to lysozyme and able to germinate efficiently. Another regulator, CotH, is controlled by CotE and is present in low amounts both during sporulation and in mature spores. In spite of this CotH controls the assembly of at least nine outer coat proteins and cooperates with CotE in producing fully resistant and efficiently germinating spores. In order to improve our understanding of CotH role in spore formation, we over-produced CotH by placing its coding region under the control of a promoter stronger than its own promoter but with a similar timing of activity during sporulation. Over-production of CotH in an otherwise wild type strain did not cause any major effect, whereas in a cotE null background a partial recovery of the phenotypes associated to the cotE null mutation was observed. Western blot, fluorescence microscopy and Surface-Enhanced Raman Scattering spectroscopy data indicate that, in the absence of CotE, over-production of CotH allowed the formation of spores overall resembling wild type spores and carrying in their coat some CotE−/CotH-dependant proteins. Our results suggest that the B. subtilis spore differentiation programme is flexible, and that an increase in the amount of a regulatory protein can replace a missing partner and partially substitute its function in the assembly of the spore coat. [ABSTRACT FROM AUTHOR]

Acanthamoeba keratitis is a rare but serious corneal disease, often observed in contact lens wearers. Clinical treatment of infected patients frequently involves the use of polyhexamethylene biguanide (PHMB), a polymer used as a disinfectant and antiseptic, which is toxic also for the epithelial cells of the cornea. Prompt and effective diagnostic tools are hence highly desiderable for both starting early therapy and timely suspension of the treatment. In this work we use Raman microspectroscopy to analyse in vitro a single Acanthamoeba cell in cystic phase. In particular, we investigate the effect of PHMB at the single-cell level, providing useful information on both the underlying biochemical mechanism and the time frame for Acanthamoeba eradication in ocular infections. Furthermore, we demonstrate that Raman spectroscopy, in conjunction with standard multivariate analysis methods, allows discriminating between live and dead Acanthamoebas, which is fundamental to optimizing patients’ treatment. [ABSTRACT FROM AUTHOR]

In this work, we establish the use of surface-enhanced Raman scattering (SERS) as a label-free analytical technique for the direct detection of G-quadruplex formation. In particular, we demonstrate that SERS analysis allows the evaluation of the relative stability of G quadruplexes that differ for the number of G tetrads and investigate several structural features of quadruplexes, such as the orientation of glycosidic bonds, the identification of distortions in the sugar-phosphate backbone, and the degree of hydrogen-bond solvation. Herein, the fluctuation of the SERS spectra, due to the specific interaction of vibrational modes with the SERS-active substrate, is quantitatively analyzed before and after quadruplex formation. The results of this study suggest a perpendicular orientation of the quadruplexes (with or without the 3'-tetra end linker) with respect to the silver colloidal surface, which opens new perspectives for the use of SERS as a label-free analytical tool for the study of the binding mode between quadruplexes and their ligands. [ABSTRACT FROM AUTHOR]

Background. A recent clinical trial showed harmful renal effects with the combined use of angiotensin-converting enzyme inhibitors (ACEI) and angiotensin-II receptor blockers (ARB) in people with diabetes or vascular disease. We examined the benefits and risks of these agents in people with albuminuria and one or more cardiovascular risk factors.Methods. MEDLINE, EMBASE and Renal Health Library were searched for trials comparing ACEI, ARB or their combination with placebo or with one another in people with albuminuria and one or more cardiovascular risk factor.Results. Eighty-five trials (21 708 patients) were included. There was no significant reduction in the risk of all-cause mortality or fatal cardiac–cerebrovascular outcomes with ACEI versus placebo, ARB versus placebo, ACEI versus ARB or with combined therapy with ACEI + ARB versus monotherapy. There was a significant reduction in the risk of nonfatal cardiovascular events with ACEI versus placebo but not with ARB versus placebo, ACEI versus ARB or with combined therapy with ACEI + ARB versus monotherapy. Development of end-stage kidney disease and progression of microalbuminuria to macroalbuminuria were reduced significantly with ACEI versus placebo and ARB versus placebo but not with combined therapy with ACEI + ARB versus monotherapy.Conclusions. ACEI and ARB exert independent renal and nonfatal cardiovascular benefits while their effects on mortality and fatal cardiovascular disease are uncertain. There is a lack of evidence to support the use of combination therapy. A comparative clinical trial with ACE, ARB and its combination in people with albuminuria and a cardiovascular risk factor is warranted. [ABSTRACT FROM PUBLISHER]

Abstract: Cell mechanical properties play an important role in determining many cellular activities. Passive microrheology techniques, such as Multiple-Particle-Tracking (MPT) give an insight into the structural rearrangements and viscoelastic response of a wide range of materials, in particular soft materials and complex fluids like cell cytoplasm in living cells. The technique finds an important field of application in large cells such as oocytes where, during their growth, several organelles and molecules are displaced in specific territories of the cell instrumental for later embryonic development. To measure cell mechanics, cells are usually deformed by many techniques that are slow and often invasive. To overcome these limits, the MPT technique is applied. Probe particles are embedded in the viscoelastic sample and their properties are extracted from the thermal fluctuation spectra measured using digital video-microscopy. The Brownian motion of a probe particle immersed in a network is directly related to the network’s mechanical properties. Particles exhibit larger motions when their local environments are less rigid or less viscous. The mean-square-displacement (MSD) of the particle’s trajectory is used to quantify its amplitude of motions over different time scales. [Copyright &y& Elsevier]

Abstract: This work reports on the application of Raman spectroscopy for the analysis of Xenopus laevis oocytes (stage-I). A two-color home-made microscope has been used for this investigation. In particular, a 785nm Raman probe has been used to acquire the spontaneous Raman scattering from the oocyte cytoplasm, while a 532nm probe has been employed to detect carotenoids through Resonant Raman Scattering. Finally, the distribution of β-carotene along a diameter of a single oocyte has been investigated. [Copyright &y& Elsevier]

This review presents the development of a Raman Tweezers system for detecting hemoglobin-related blood disorders at a single cell level. The study demonstrates that the molecular fingerprint insight provided by Raman analysis holds great promise for distinguishing between healthy and diseased cells in the field of biomedicine. Herein a Raman Tweezers system has been applied to investigate the effects of thalassemia, a blood disease quite diffuse in the Mediterranean Sea region. By resonant excitation of hemoglobin Raman bands, we examined the oxygenation capability of normal, alpha- and beta-thalassemic erythrocytes. A reduction of this fundamental red blood cell function, particularly severe for beta-thalassemia, has been found. Raman spectroscopy was also used to draw hemoglobin distribution inside single erythrocytes; the results confirmed the characteristic anomaly (target shape), occurring in thalassemia and some other blood disorders. The success of resonance Raman spectroscopy for thalassemia detection reported in this review provide an interesting starting point to explore the application of a Raman Tweezers system in the analysis of several blood disorders. [ABSTRACT FROM AUTHOR]

The aim of this work is to investigate if the optical tweezers (OT) are suitable as a diagnostic tool for monitoring the oligodeoxyribonucleotide (ODN) structural behavior in solution. Preliminary experiments, performed on the quadruplex formed by the ODN sequence TGGGGT, showed that the OT can be used as a probe for ODN structuration by monitoring the medium viscosity changes associated with ODN folding-unfolding processes. [ABSTRACT FROM AUTHOR]

Abstract: In this work we discuss on a compact spectrometer based on DFB diode lasers for detection of chloridric and fluoridric acids. HCl and HF concentrations are determined through optical absorption of the P(4) line (λ =1.7μm) and the R(3) line (λ =1.3μm), respectively. Both lines belong to first overtone vibrational bands and their linestrengths are 7.8×10−21 cm/molecule for HCl and 2.8×10−20 cm/molecule for HF. We chose these lines for their relative high intensities and because they are quite far from water vapour lines which represent the main interfering gas for trace-gases analysis. To detect these species we used two different high frequency modulation techniques: two-tone frequency modulation spectroscopy (f 1 =800MHz and f 2 =804MHz) was used for HCl while for HF we followed a simpler approach based on wavelength modulation spectroscopy (f =600kHz). We demonstrate that the two techniques provide comparable detection limit of about 80ppbV at atmospheric pressure. Positive testing of our spectrometer makes it suitable for in situ measurements of exhaust gases coming from waste incinerators. [Copyright &y& Elsevier]

We have studied the mechanical properties of the cytoplasm of starfish oocytes before and after maturation using the multiple particle tracking technique. Fluorescent microbeads are microinjected in oocytes of Astropecten aranciacus starfishes. Mechanical properties are obtained analyzing the mean-square displacement (MSD) of the beads trajectories. The ensemble-averaged MSD provides information about diffusion coefficient D, and the cytoplasm viscosity η is estimated through the Stokes–Einstein equation. Results show that maturation is correlated with change of cytoplasm viscosity. The results can be ascribed to F-actin reorganization as also supported by fluorescence confocal microscopy. [ABSTRACT FROM AUTHOR]

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper. [ABSTRACT FROM AUTHOR]

Sensitive detection of altered proteins expression in plasma membranes is of fundamental importance, for both diagnostic and prognostic purposes. Surface-Enhanced Raman Scattering (SERS) has proven to be a quite sensitive approach to detect proteins, even in very diluted samples. However, proteins detection in complex environment, such as the cellular membrane, is still a challenge. Herein, we demonstrate a SERS-based platform to reveal the overexpression of target proteins in cell membranes. As a proof of concept, we implemented ectopic expression of carbonic anhydrase IX (CA IX) and epidermal growth factor receptor (EGFR) in the plasma membrane of the SKOV3 tumor cell line. Our outcomes demonstrate that SERS signals from cells put in contact with a hyperuniform SERS substrate allow highlighting subtle differences in the biochemical composition of cell membranes, normally hidden in spontaneous Raman confocal microscopy. This opens new opportunities for a label-free membrane analysis and bio-sensing in a broader sense. [ABSTRACT FROM AUTHOR]

We experimentally measure the effects of fluid inertia on the diffusion of a Brownian particle at very long time scales. In previous experiments, the use of standard optical tweezers introduced a cutoff in the free diffusion of the particle, which limited the measurement of these effects to times comparable with the relaxation time of the fluid inertia, i.e., a few milliseconds. Here, by using blinking optical tweezers, we detect these inertial effects on time scales several orders longer up to a few seconds. The measured mean square displacement of a freely diffusing Brownian particle in a liquid shows a deviation from the Einstein-Smoluchowsky theory that diverges with time. These results are consistent with a generalized theory that takes into account not only the particle inertia but also the inertia of the surrounding fluid. [ABSTRACT FROM AUTHOR]

In cystic fibrosis (CF) patients airways mucus shows an increased viscoelasticity due to the concentration of high molecular weight components. Such mucus thickening eventually leads to bacterial overgrowth and prevents mucus clearance. The altered rheological behavior of mucus results in chronic lung infection and inflammation, which causes most of the cases of morbidity and mortality, although the cystic fibrosis complications affect other organs as well. Here, we present a quantitative study on the correlation between cystic fibrosis mucus viscoelasticity and patients clinical status. In particular, a new diagnostic parameter based on the correlation between CF sputum viscoelastic properties and the severity of the disease, expressed in terms of FEV1 and bacterial colonization, was developed. By using principal component analysis, we show that the types of colonization and FEV1 classes are significantly correlated to the elastic modulus, and that the latter can be used for CF severity classification with a high predictive efficiency (88%). The data presented here show that the elastic modulus of airways mucus, given the high predictive efficiency, could be used as a new clinical parameter in the prognostic evaluation of cystic fibrosis. [ABSTRACT FROM AUTHOR]