Cell‐Instructive Microgels with Tailor‐Made Physicochemical Properties

A microfluidic in vitro cell encapsulation platform to systematically test the effects of microenvironmental parameters on cell fate in 3D is developed. Multiple cell types including fibroblasts, embryonic stem cells, and cancer cells are incorporated in enzymatically cross‐linked poly(ethylene glycol)‐based microgels having defined and tunable mechanical and biochemical properties. Furthermore, different approaches to prevent cell “escape” from the microcapsules are explored and shown to substantially enhance the potential of this technology. Finally, coencapsulation of microgels within nondegradable gels allows cell viability, proliferation, and morphology to be studied in different microenvironmental conditions up to two weeks in culture.     

Metformin Protects against Radiation-Induced Acute Effects by Limiting Senescence of Bronchial-Epithelial Cells

Radiation-induced damage to normal lung parenchyma remains a dose-limiting factor in thorax-associated radiotherapy (RT). Severe early and late complications with lungs can increase the risk of morbidity in cancer patients after RT. Herein, senescence of lung epithelial cells following RT-induced cellular stress, or more precisely the respective altered secretory profile, the senescence-associated secretory phenotype (SASP), was suggested as a central process for the initiation and progression of pneumonitis and pulmonary fibrosis. We previously reported that abrogation of certain aspects of the secretome of senescent lung cells, in particular, signaling inhibition of the SASP-factor Ccl2/Mcp1 mediated radioprotection especially by limiting endothelial dysfunction. Here, we investigated the therapeutic potential of a combined metformin treatment to protect normal lung tissue from RT-induced senescence and associated lung injury using a preclinical mouse model of radiation-induced pneumopathy. Metformin treatment efficiently limited RT-induced senescence and SASP expression levels, thereby limiting vascular dysfunctions, namely increased vascular permeability associated with increased extravasation of circulating immune and tumor cells early after irradiation (acute effects). Complementary in vitro studies using normal lung epithelial cell lines confirmed the senescence-limiting effect of metformin following RT finally resulting in radioprotection, while fostering RT-induced cellular stress of cultured malignant epithelial cells accounting for radiosensitization. The radioprotective action of metformin for normal lung tissue without simultaneous protection or preferable radiosensitization of tumor tissue might increase tumor control probabilities and survival because higher radiation doses could be used.     

Cation exchange in ZnSe nanocrystals for signal amplification in bioassays

ZnSe nanocrystals (NCs), possessing low native luminescence but high biocompatibility, were employed as labeling tags in bioassays. They were able to amplify each target recognition event thousands of times through a cation-exchange reaction (CXAmp) that released over 3000 encapsulated Zn(2+) from one single NC. The freed cations in turn triggered strong fluorescence from the Zn-responsive dyes. The present study demonstrated that CXAmp with ZnSe delivered superior detection performance in comparison to the conventional labeling methods. The overall fluorescence intensity of CXAmp using 5 nM ZnSe NCs was 30 times higher than that from 5 nM core-shell CdSe/ZnS quantum dots (QDs). The limit of detection (LOD) obtained with ZnSe-based CXAmp was 10-fold lower than with horseradish peroxidase (HRP) labeling, and the detection sensitivity, represented by the slope of the signal-versus-concentration curve, was 20-fold higher. When applied to detect immunoglobulin E (IgE) in a sandwich format, a LOD of 1 ng/mL was achieved. The highly sensitive CXAmp also allowed detection of the total IgE content in dilute human serum, in which the abundant matrix proteins exhibited less interference and more accurate quantification could be performed. Besides high signal amplification efficiency and good biocompatibility, CXAmp with ZnSe could be easily adapted to common laboratory settings and act as a universal labeling system for reliable detection of low-abundance targets.     

Identifying a distinctive familial frequency band in reaction time fluctuations in ADHD.

Objective: Patients with ADHD are typically more variable in their reaction times (RT) than control children. Signal processing analyses have shown that time series RT data of children with ADHD have a distinctive low frequency periodic structure suggestive of a pattern of occasional spontaneous performance lapses. Here we use a fine-grained analysis of spectral power across a broader frequency range to differentiate the periodic qualities of ADHD time series RT data from (a) 1/frequency noise, and (b) control performance. We also assess the familiality of these frequencies by using a proband-sibling design. Method: Seventy-one children with ADHD, one of their siblings, and 50 control participants completed a simple RT task. Power across the RT frequency spectrum was calculated. The frequencies significantly differentiating the two groups were identified. Familiality was assessed in two ways: first, by comparing probands with their unaffected siblings and controls, and, second, by investigating the siblings of neuropsychologically impaired and unimpaired children with ADHD. Results: Analyses converged to highlight the potential importance of the .20–.26 Hz band in differentiating the periodic structure of ADHD RT time series data from both 1/frequency noise and control performance. This frequency band also showed the strongest evidence of familiality. Conclusions: RT performance of children with ADHD had a distinctive periodic structure. The band identified as most differentiating and familial was at a higher frequency than in most previous reports. This highlights the importance of employing tasks with faster interstimulus intervals that will allow a larger portion of the frequency spectrum to be examined. (PsycINFO Database Record (c) 2011 APA, all rights reserved)     

The face in the crowd effect unconfounded: Happy faces, not angry faces, are more efficiently detected in single- and multiple-target visual search tasks.

Is it easier to detect angry or happy facial expressions in crowds of faces? The present studies used several variations of the visual search task to assess whether people selectively attend to expressive faces. Contrary to widely cited studies (e.g., ?hman, Lundqvist, & Esteves, 2001) that suggest angry faces “pop out” of crowds, our review of the literature found inconsistent evidence for the effect and suggested that low-level visual confounds could not be ruled out as the driving force behind the anger superiority effect. We then conducted 7 experiments, carefully designed to eliminate many of the confounding variables present in past demonstrations. These experiments showed no evidence that angry faces popped out of crowds or even that they were efficiently detected. These experiments instead revealed a search asymmetry favoring happy faces. Moreover, in contrast to most previous studies, the happiness superiority effect was shown to be robust even when obvious perceptual confounds—like the contrast of white exposed teeth that are typically displayed in smiling faces—were eliminated in the happy targets. Rather than attribute this effect to the existence of innate happiness detectors, we speculate that the human expression of happiness has evolved to be more visually discriminable because its communicative intent is less ambiguous than other facial expressions. (PsycINFO Database Record (c) 2011 APA, all rights reserved)     

pH dependent spontaneous growth of ellagic acid assemblies for targeting HeLa cells

Herein, we have studied the self-assembly and the spontaneous growth of microassemblies of the plant polyphenol ellagic acid for HeLa cancer cell imaging and therapy. The growth of the assemblies was studied at varying pH over time. It was found that initially microspheres were formed which gradually transformed into microfibers via nucleation and polymerization process. The optimum growth of microfibers was found to be in the pH range of 6-8. We have shown that the microfibers successfully adhered to the HeLa cell membranes and inhibited their proliferation. This biological approach, using assemblies derived from plant polyphenols, may be used for direct cellular drug delivery and may potentially help develop a simple and economical method to create building blocks with desired properties for a new generation of sensors, bioimaging and drug delivery systems.     

Flavonoids and furocoumarins distribution of unripe chinotto (<Emphasis Type="Italic">Citrus</Emphasis> × <Emphasis Type="Italic">myrtifolia Rafinesque</Emphasis>) fruit: beverage processing homogenate and juice characterization

The present study reports for the first time qualitative and quantitative data about flavonoids and furocoumarins distribution of unripe Citrus × myrtifolia Rafinesque homogenate, used as ingredient of the popular soft drink Chinotto. No data are available in literature about the chemical characterization of flavonoids and furocoumarins of the chinotto homogenate used to produce the industrial extract. Samples for analysis were therefore processed to better reproduce the crude materials used to obtain industrial chinotto extract: the whole fruits were cut in half, seeds removed, softly squeezed discarding part of the juice, and homogenized in small pieces. The polyphenolics separation was made by HPLC–PDA detector coupled to ESI/MS/MS in positive and negative mode. It has allowed the identification and relative quantification of 16 compounds, among which the flavone-C-glycoside lucenin-2, the 3-hydroxy-3-methylglutaryl flavanone glycoside melitidin and five polymethoxylated flavones have been detected for the first time in Citrus × myrtifolia. The flavonoids and furocoumarins distribution was compared with one of the Citrus × myrtifolia juice, and the antioxidant activity of both samples was assessed by ORAC (Oxygen Radical Absorbance Capacity)-fluorescein assay.     

Quantitation of Saccharin and Cyclamate in Tabletop Formulations by Portable Raman and NIR Spectrometers in Combination with Partial Least Squares Regression

Rapid, direct, and reagent-free screening tools using vibrational spectroscopy in combination with partial least squares regression (PLSR) were developed for the determination of sodium saccharin and sodium cyclamate in tabletop formulations. The four vibrational spectroscopic instruments employed were a portable Raman spectrometer, a NIR handheld device, and two FT-NIR benchtop spectrometers. Wavenumber ranges and type of spectral pretreatment were optimized for each PLSR calibration model using an independent validation set. Each sweetener model provided reliable predictions (low errors in validation and r ² above 0.90) for both saccharin and cyclamate samples. Optimized models were tested with four commercially available tabletop formulations in order to simulate the application of the developed models towards routine sweetener analysis. With the exception of one model, the sweetener concentration predictions in commercial tabletop formulations using the portable devices were not significantly different from those based on spectra collected on the benchtop spectrometers. PLSR-predicted mean sweetener concentrations were within 80–120% of the label declared values, while the sweetener without a label declaration had consistent concentrations across the analytical methods used. As shown by the good agreement between spectroscopic and chromatographic analyses, the portable spectrometers offer an alternative to traditional chromatographic methods. To our knowledge, this is the first time portable Raman and handheld NIR devices with PLSR calibration models have been employed to evaluate sweeteners, and these analytical methods hold potential to be used for rapid screening of tabletop formulations for quality assurance and for regulatory labeling verification.