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Chronic pain conditions are frequently comorbid with anxiety problems. The controlled diaphragmatic breathing technique has been applied in both conditions. However, few studies have examined its efficacy in reducing perceived pain and state anxiety in the population with both pathologies. Thus, the study objective was to investigate the immediate effects of a single-session diaphragmatic breathing intervention on perceived pain and state anxiety in a group of people with chronic pain. We analyzed the influence of sociodemographic variables, dispositional anxiety, average pain during the last week, respiratory rate, and inhalation/exhalation ratio. The sample comprised 169 people with chronic pain from Spain and Poland who were evaluated before and after performing an online controlled diaphragmatic breathing task. The Wilcoxon test showed significant decreases in state anxiety and perceived pain. Linear regression analyses revealed an association between dispositional anxiety and pretest/posttest differences in state anxiety, as well as an association between both average pain in the last week and nationality and pretest/posttest differences in perceived pain. We conducted an exploratory analysis of the differences between both countries in the study variables. We discuss the results and the relevance of further studies on the efficacy of this technique in reducing perceived pain and state anxiety. [ABSTRACT FROM AUTHOR]

Purpose: Oocytes from women presenting primary ovarian insufficiency (POI) generate viable embryos at a lower rate than non-POI women, but the mechanisms responsible for the lower oocyte quality remain elusive. Due to the scarcity of human oocytes for research, animal models provide a promising way forward. We aimed at investigating the molecular events characterizing final maturation in POI oocytes in a well-defined POI-like bovine model. Methods: Single-cell RNA-sequencing of bovine control and POI-like, GV, and MII oocytes (n = 5 per group) was performed. DEseq2 was used to identify differentially expressed genes. Further, a Gene set enrichment analysis and a transcriptomic meta-analysis between bovine and human oocytes were performed. Results: In control cows, we found 2223 differentially expressed genes between the GV and MII stages. Specifically, the affected genes were related to RNA processing and transport, protein synthesis, organelle remodeling and reorganization, and metabolism. The meta-analysis with a set of young human oocytes at different maturation stages revealed 315 conserved genes through the GV-MII transition in cows and humans, mostly related to meiotic progression and cell cycle. Gene expression analysis between GV and MII of POI-like oocytes showed no differences in terms of differentially expressed genes, pointing towards a substantial failure to properly remodel the transcriptome in the POI model, and with the clustering analysis indicating that the cow's genetic background had a higher impact than the oocyte's maturation stage. Conclusion: Overall, we have identified and characterized a valuable animal model of POI, paving the way to identifying new molecular mechanisms involved in POI. [ABSTRACT FROM AUTHOR]

Empathy in healthcare professionals is associated with better treatment outcomes and higher satisfaction among patients with chronic pain. Activity patterns play an essential role in the adjustment of these patients and, as a pain behaviour, may have a communicative function and elicit distinct empathic responses. This study investigated whether the activity pattern profiles characteristic of these patients had differential effects on the empathic response (empathic distress and compassion/sympathy) of future healthcare professionals. Healthcare professionals should improve their knowledge about the role of different activity patterns in the well-being of people with chronic pain and receive specific training in empathic skills. We controlled for several variables that could affect the empathic response (sex, age, academic degree, previous experience of chronic pain, and dispositional empathy). A total of 228 undergraduates performed an experimental task using vignettes depicting four activity pattern profiles displayed by people with chronic pain and completed questionnaires measuring dispositional and situational empathy. We conducted a MANCOVA analysis. Undergraduates showed more compassion/sympathy toward the medium cycler profile than toward the doer profile. Participants' age was associated with empathic distress. Sex, academic degree, and previous experiences with chronic pain were not associated with their empathic response to the vignettes. Dispositional perspective-taking and empathic concern were significantly associated with compassion/sympathy responses, and personal distress was significantly associated with empathic distress. Activity pattern profiles may have a communicative function and elicit different empathic responses toward people with chronic pain. Individual differences in dispositional empathy play an important role on situational empathic responses. [ABSTRACT FROM AUTHOR]

In this study, an exhaustive chemical characterization of a Dunaliella salina (DS) microalga extract obtained using supercritical fluids has been performed, and its neuroprotective capacity has been evaluated in vivo using an Alzheimer's disease (AD) transgenic model of Caenorhabditis elegans (strain CL4176). More than 350 compounds were annotated in the studied DS extract, with triacylglycerols, free fatty acids (FAs), carotenoids, apocarotenoids and glycerol being the most abundant. DS extract significantly protects C. elegans in a dose-dependent manner against Aβ-peptide paralysis toxicity, after 32 h, 53% of treated worms at 50 µg/mL were not paralyzed. This concentration was selected to further evaluate the transcriptomics and metabolomics changes after 26 h by using advanced analytical methodologies. The RNA-Seq data showed an alteration of 150 genes, mainly related to the stress and detoxification responses, and the retinol and lipid metabolism. The comprehensive metabolomics and lipidomics analyses allowed the identification of 793 intracellular metabolites, of which 69 were significantly altered compared to non-treated control animals. Among them, different unsaturated FAs, lysophosphatidylethanolamines, nucleosides, dipeptides and modified amino acids that have been previously reported as beneficial during AD progression, were assigned. These compounds could explain the neuroprotective capacity observed, thus, providing with new evidences of the protection mechanisms of this promising extract. [ABSTRACT FROM AUTHOR]

This study presents a novel vision of the brown seaweed Durvillaea antarctica aka 'cochayuyo' as an important food commonly consumed in Chile. ´Cochayuyo´ is a rich source of antioxidant and neuroprotective activity compounds demonstrated by evaluating the bioactivity associated with conventional and pressurized liquid extracts. Variables such as extraction yield, DPPH antioxidant assay, TEAC, and acetylcholinesterase (AChE) inhibitory capacity were studied employing Central Composite Design (CCD) based on a response surface methodology (RSM) and Pareto charts. Firstly, solvent polarity was evaluated by conventional extraction for the best antioxidant activity and they were employed in the experimental design (water/ethanol variations). Then, factors such as temperature (40–180 °C) and solvents (water/ethanol in 0, 50 to 100%) were selected resulting in CCD (12 total runs). In general, pressurized liquid extraction (PLE) improved the extraction yield and antioxidant activities compared to conventional extraction (twofold more using water). Moreover, PLE results showed that a higher yield (63.28%), was obtained with 50:50% water/ethanol at 180 °C. However, DPPH antioxidant assay was improved at the same temperature under 100% ethanol (EC50 equal to 0.16 mg mL−1). For the neuroprotective activity, D. antarctica extracts were enriched under low-intermediate temperature (40–110 °C) and 100% water (IC50 equal to 148.62 μg mL−1). After multiple response optimization, the best conditions to obtain antioxidant and neuroprotective extracts with high yield were 180 ºC and a ratio of 71:29 water:ethanol as extraction solvent. The results indicated that D. antarctica could be considered a functional food with significant antioxidant benefits and neuroprotective effects on human health. [ABSTRACT FROM AUTHOR]

Alzheimer's disease (AD) is the most common form of dementia caused by a progressive loss of neurons from different regions of the brain. This multifactorial pathophysiology has been widely characterized by neuroinflammation, extensive oxidative damage, synaptic loss, and neuronal cell death. In this sense, the design of multi-target strategies to prevent or delay its progression is a challenging goal. In the present work, different in vitro assays including antioxidant, anti-inflammatory, and anti-cholinergic activities of a carotenoid-enriched extract from Dunaliella salina microalgae obtained by supercritical fluid extraction are studied. Moreover, its potential neuroprotective effect in the human neuron-like SH-SY5Y cell model against remarkable hallmarks of AD was also evaluated. In parallel, a comprehensive metabolomics study based on the use of charged-surface hybrid chromatography (CSH) and hydrophilic interaction liquid chromatography (HILIC) coupled to high-resolution tandem mass spectrometry (Q-TOF MS/MS) was applied to evaluate the effects of the extract on the metabolism of the treated cells. The use of advanced bioinformatics and statistical tools allowed the identification of more than 314 metabolites in SH-SY5Y cells, of which a great number of phosphatidylcholines, triacylglycerols, and fatty acids were significantly increased, while several phosphatidylglycerols were decreased, compared to controls. These lipidomic changes in cells along with the possible role exerted by carotenoids and other minor compounds on the cell membrane might explain the observed neuroprotective effect of the D. salina extract. However, future experiments using in vivo models to corroborate this hypothesis must be carried out. [ABSTRACT FROM AUTHOR]

The study of the phytochemical composition of seed oils is of upmost importance for the food and cosmetic industries, mainly considering their associated biological properties. Extraction of seed oils using supercritical fluids (SFE) is an ecological and green alternative to conventional extraction processes since it is able to provide with potent bioactive extracts, avoiding degradation and transformation of the compounds present originally in the raw material. The objective of the present work was the extraction of pure fractions of polar lipids and their chemical characterization using chromatographic techniques such as GC-MS and LC-DAD-MS/MS of blackberry (Rubus glaucus) and passion fruit (Passiflora edulis) seed oils extracted by supercritical carbon dioxide. Oleamides derived from oleic acid were identified as the main compounds in both samples; in particular, 9-octadecenamide was the major identified oleamide. Besides, the extract obtained from passion fruit showed to be a source of linoleic acid, while the SFE extract from blackberry presented important concentrations of vanillin. The chemical composition of these seed oils can be of high interest for their further use in cosmetics and food industry. [ABSTRACT FROM AUTHOR]

Purpose: This study aimed to determine the role of Wnt pathway in mouse embryonic stem cell (mESC) derivation from single blastomeres isolated from eight-cell embryos and in the pluripotency features of the mESC established. Methods: Wnt activator CHIR99021, Wnt inhibitor IWR-1-endo, and MEK inhibitor PD0325901 were used alone or in combination during ESC derivation and maintenance from single blastomeres biopsied from eight-cell embryos. Alkaline phosphatase activity, FGF5 levels, expression of key pluripotency genes, and chimera formation were assessed to determine the pluripotency state of the mESC lines. Results: Derivation efficiencies were highest when combining pairs of inhibitors (15–24.7%) than when using single inhibitors or none (1.4–10.1%). Full naïve pluripotency was only achieved in CHIR- and 2i-treated mESC lines, whereas IWR and PD treatments or the absence of treatment resulted in co-existence of naïve-like and primed-like pluripotency features. IWR + CHIR- and IWR + PD-treated mESC displayed features of primed pluripotency, but IWR + CHIR-treated lines were able to generate germline-competent chimeric mice, resembling the predicted properties of formative pluripotency. Conclusion: Wnt and MAPK pathways have a key role in the successful derivation and pluripotency features of mESC from single precompaction blastomeres. Modulation of these pathways results in mESC lines with various degrees of naïve-like and primed-like pluripotency features. [ABSTRACT FROM AUTHOR]

Purpose: The quality of life of individuals with chronic musculoskeletal pain partly depends on their capacity to adjust their personal goals. Vignettes have been rarely used to assess this ability. Therefore, this study aimed to test the relationships between vignettes assessing different goal strategies and chronic pain adaptation (i.e., daily functioning, pain-related impairment, and psychological well-being). Methods: The sample comprised 258 individuals with chronic musculoskeletal pain who completed a series of questionnaires and vignettes. The vignettes presented a short description of a situation in which a person with chronic pain experienced a threat to a valued domain-specific goal and had to choose a possible goal management solution (i.e., goal persistence, flexibility reengagement, and disengagement). Hierarchical regression analyses were used to predict chronic pain adaptation using the selected vignette strategies as predictors. Results: After controlling for age, sex, pain intensity, and the responses to the dispositional goal management scales, persistence, reengagement, and disengagement goal strategies presented in the case scenarios predicted daily functioning (p <.001). Persistence, flexibility, disengagement (p <.001), and reengagement (p <.05) predicted pain-related impairment. Persistence, disengagement (p <.001), and flexibility (p <.05) predicted psychological well-being scores. Conclusion: The use of vignettes could be useful to assess goal adjustment because this methodology enables respondents to provide more context-specific responses. The results of this approach could be used to improve clinical practice aimed at helping people with chronic musculoskeletal pain to better cope with this health condition. [ABSTRACT FROM AUTHOR]

The green microalga Haematococcus pluvialis has been widely studied due to its capacity to accumulate great amounts of astaxanthin, a high-value carotenoid with biological activities. In the present work, two green compressed fluid–based processes, pressurized liquid extraction (PLE) and supercritical antisolvent fractionation (SAF), are integrated to obtain an astaxanthin-enriched extract from this microalga. PLE was carried out using pressurized ethanol as solvent, for 20 min, at 10 MPa, and 50 °C as extraction temperature. Subsequently, the obtained extract was processed by SAF to further purify the carotenoid fraction. The SAF process was optimized using a 3-level factorial experimental design and considering three experimental variables: (i) CO2 pressure (10–30 MPa), (ii) percentage of water in the PLE extract (20–50%), and (iii) PLE extract/supercritical-CO2 flow rate ratio (0.0125–0.05). Total carotenoid content was evaluated in both extracts and raffinates. Best results were obtained at 30 MPa, 0.05 feed/SC-CO2 mass flow rate, and 20% (v/v) of water in the feed solution, achieving values of 120.3 mg g−1 carotenoids in extract (in the SAF extract fraction), which were significantly higher than those obtained in the original PLE extract. In parallel, a new fast two-dimensional comprehensive liquid chromatography (LC×LC) method was optimized to get the full carotenoid profile of these extracts in less than 25 min. This is the first time that the use of a C30 column is reported in an on-line LC×LC system. [ABSTRACT FROM AUTHOR]

Purpose: To assess the role of the genetic background, the culture medium supplements, and the presence of modulators of signaling pathways on mouse embryonic stem cell derivation from single blastomeres from 8-cell embryos.Methods: Mice from permissive and non-permissive genetic backgrounds, different culture media supplements, knockout serum replacement (KSR) and N2B27, and the presence or absence of 2i treatment were used to derive mouse embryonic stem cells (mESC) from single blastomeres isolated from 8-cell embryos and from control embryos at the blastocyst stage. After the sixth passage, the putative mESC were analyzed by immunofluorescence to assess their pluripotency and, after in vitro differentiation induction, their ability to differentiate into derivatives of the three primary germ layers. Selected mESC lines derived from single blastomeres in the most efficient culture conditions were further characterized to validate their stemness.Results: In control embryos, high mESC derivation efficiencies (70-96.9%) were obtained from permissive backgrounds or when embryos were cultured in medium complemented with 2i regardless of their genetic background. By contrast, only blastomeres isolated from embryos from permissive background cultured in KSR-containing medium complemented with 2i were moderately successful in the derivation of mESC lines (22.9-24.5%). Moreover, we report for the first time that B6CBAF2 embryos behave as permissive in terms of mESC derivation.Conclusions: Single blastomeres have higher requirements than whole blastocysts for pluripotency maintenance and mESC derivation. The need for 2i suggests that modulation of signaling pathways to recreate a commitment towards inner cell mass could be essential to efficiently derive mESC from single blastomeres. [ABSTRACT FROM AUTHOR]

Purpose: The purpose of this study was to evaluate the longitudinal trajectories of acute stress reactions over the course of diagnosis, treatment, and follow-up assessments in a group of non-metastatic breast cancer patients during five different moments of the illness process, and to identify psychological predictors of the trajectories.Methods: The sample was formed by 102 non-metastatic breast cancer patients treated with adjuvant chemotherapy. Latent growth mixture models (LGMM) were used to identify latent classes, and we used multinomial logistic regression in a conditional model to examine predictors to differentiate between trajectories.Results: We identified four different groups according to their trajectories: (1) a resilient group, (44.5 %); (2) a mild acute stress group, (40.6 %); (3) a delayed-recovery group (11.9 %); and (4) a chronic acute stress group (2.9 %). Moreover, anxious preoccupation showed the strongest significant effects in predicting each class, whereas cognitive avoidance and type C personality had moderate effects for participants in the mild acute stress group.Conclusions: This study demonstrates that the majority of breast cancer patients in our study were resilient, with only a small percentage showing chronic acute stress. Because coping strategies, specifically anxious preoccupation, and not more stable variables played a main role in the prediction of acute stress trajectories, future preventive interventions should center in promoting more adaptive coping strategies in breast cancer patients. [ABSTRACT FROM AUTHOR]

Purpose: International organizations such as National Comprehensive Cancer Network and NICE recommend implementation of routine screening programs for detecting and managing psychosocial distress among cancer patients. The selection of an adequate screening tool is crucial to the effectiveness of these programs. The present study examines the emotional symptomatology captured by the Distress Thermometer (DT) and its accuracy and validity as a screening tool in cancer. It also explores the possible discrepancy between patient distress and the use of psycho-oncology resources. Methods: A heterogeneous sample of 962 adult cancer patients completed the DT, the Problem List (PL), the Brief Symptom Inventory-18 (BSI-18), and the Psychosocial Questionnaire. Results: The DT was significantly correlated with BSI-18 symptoms and the emotional problems listed on the PL. Receiver Operating Characteristic analysis showed good diagnostic accuracy for the DT (area under the curve = .82, 95 %CI [.79-.85]). For a selected DT cutoff of 5, standard measures (sensitivity = 90 %, specificity = 64 %, predictive positive value = 35 %, and negative predictive value = 97 %) and Clinical Utility Indexes (utility index negative = .62 and utility index positive = .32) indicated that the DT was adequate for 'screening' while it was limited for 'case finding.' Finally, 81.30 % of patients with clinical distress had not sought or were not receiving professional psychosocial support at the time of the study. Conclusion: The DT is appropriate for use as a rapid screening instrument for cancer patients in a Spanish population because it assesses a broad concept of distress including both anxiety and depression symptoms. The diagnostic accuracy of the DT could be improved with minor proposed modifications to the DT and the inclusion of nonemotional ultrashort measures. [ABSTRACT FROM AUTHOR]

Individual tagging of oocytes and embryos through the attachment of micrometer-sized polysilicon barcodes to their zona pellucida (ZP) is a promising approach to ensure their correct identification and traceability in human assisted reproduction and in animal production programs. To provide barcodes with the capacity of binding to the ZP, they must be first biofunctionalized with a biomolecule capable of binding to the ZP of both oocytes and embryos. The aim of this work was to select, among an anti-ZP2 antibody and the two lectins wheat germ agglutinin (WGA) and phytohemagglutinin-L, the most optimal biomolecule for the eventual biofunctionalization of barcodes, using mouse oocytes and embryos and commercially available microspheres as a model. Despite the anti-ZP2 antibody showed the highest number of binding sites onto the ZP surface, as determined by field emission scanning electron microscopy, the binding of anti-ZP2-biofunctionalized microspheres to the ZP of cultured oocytes and embryos was less robust and less stable than the binding of lectin-biofunctionalized ones. WGA proved to be, among the three candidates tested, the most appropriate biomolecule to biofunctionalize microparticles with the aim to attach them to the ZP of both oocytes and embryos and to maintain them attached through oocyte activation (zona reaction) and in vitro culture up to the blastocyst stage. As saccharides recognized by WGA are highly abundant in the ZP of most mammalian species, WGA-biofuncionalized microparticles would be able to attach to the ZP of oocytes/embryos of species other than the mouse, such as humans and farm animals. [ABSTRACT FROM AUTHOR]

A novel experimental design was used to optimize the extraction of carotenoids from Neochloris oleoabundans using pressurized liquid extraction with food-grade solvents such as ethanol and limonene. Experimental factors, including the extraction temperature and the solvent composition, were optimized using a three-level factorial design. The response variables extraction yield and total amount of carotenoids were assessed. The statistical analysis of the results provided mathematical models to predict the behavior of the responses as a function of the factors involved in the process. The optimum conditions predicted by the model developed in this study were 112 °C as the extraction temperature and 100 % ethanol as the extraction solvent. Chemical characterization of the extracts obtained was performed by means of high-performance liquid chromatography-tandem mass spectrometry. The results obtained demonstrated that, under certain growth conditions (photoautotrophically cultured in a medium supplemented with 0.3 g L KNO), N. oleoabundans accumulated significant total amounts of the carotenoids (from 57.4 to 120.2 mg carotenoids per gram of extract depending on the extraction conditions), mainly lutein, cantaxanthin, zeaxanthin, and astaxanthin monoesters and diesters. [ABSTRACT FROM AUTHOR]

In this work, the development and optimization of a new methodology to analyze grape seed procyanidins based on the application of two-dimensional comprehensive LC is presented. This two-dimensional method involves the use of a microbore column containing a diol stationary phase in the first dimension coupled to either a C partially porous short column or a C monolithic column in the second dimension. The orthogonal hydrophilic interaction × reversed phase liquid chromatography (HILIC×RP-LC) system is interfaced through a ten-port two-position switching valve. The optimized HILIC×RP-LC separation followed by diode array and tandem mass spectrometry detection (HILIC×RP-LC-DAD-MS/MS) made possible the direct analysis of a complex grape seed extract and allowed the tentative identification of 43 flavan-3-ols, including monomers and procyanidin oligomers till a polymerization degree of 7 units with different galloylation degrees. To the best of our knowledge, this is the first time that this powerful analytical technique is employed to characterize complex procyanidin samples. This work successfully demonstrates the great capabilities of the HILIC×RP-LC-DAD-MS/MS coupling for the direct analysis of very complex natural samples like grape seeds. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]

Organoselenium compounds have already been reported to be good anticarcinogenic candidates. A new selenoquinazoline derivative, 2,4-bis(selenomethyl)quinazoline (compound 1), has been synthesized, spectroscopically characterized and its crystal structure has been studied. An intermolecular coupling between C and $$ {\text{H}}_{5}^{\prime } $$ in the Heteronuclear Multiple Bond Correlation (HMBC) experiment has been observed. Assuming that the head-to-tail overlap of parallel molecules (as identified by X-ray diffraction) remains in solution to give bimolecular entities, the π-π interaction enables heteronuclear coupling between the former atoms with a three-bond distance [C···(π-π)··· $$ {\text{C}}_{5}^{\prime } $$- $$ {\text{H}}_{5}^{\prime } $$]. The crystal structure of compound 1 has been solved by X-ray diffraction. It crystallizes in triclinic system, space group P−1. Unit cell parameters are a = 7.4969(7) Å, b = 8.7008(8) Å, c = 10.1666(9) Å, α = 110.215(2)°, β = 90.354(2)°, γ = 115.017(1)°. Linear chains in crystals of compound 1 are generated by C-H···Se and Se···Se bonds between molecules. Furthermore, head-to-tail overlap of parallel molecules, in which π-π interactions can occur, is observed. Compound 1 exhibited a cytotoxic effect in all of the evaluated tumoral cell lines and showed a higher cytotoxic effect in colon and breast cancer cell lines than etoposide, which was used as a reference compound. [ABSTRACT FROM AUTHOR]

Efforts to efficiently derive embryonic stem cells (ESC) from isolated blastomeres have been done to minimize ethical concerns about human embryo destruction. Previous studies in our laboratory indicated a poor derivation efficiency of mouse ESC lines from isolated blastomeres at the 8-cell stage (1/8 blastomeres) due, in part, to a low division rate of the single blastomeres in comparison to their counterparts with a higher number of blastomeres (2/8, 3/8 and 4/8 blastomeres). Communication and adhesion between blastomeres from which the derivation process begins could be important aspects to efficiently derive ESC lines. In the present study, an approach consisting in the adhesion of a chimeric E-cadherin (E-cad-Fc) to the blastomere surface was devised to recreate the signaling produced by native E-cadherin between neighboring blastomeres inside the embryo. By this approach, the division rate of 1/8 blastomeres increased from 44.6% to 88.8% and a short exposure of 24 h to the E-cad-Fc produced an ESC derivation efficiency of 33.6%, significantly higher than the 2.2% obtained from the control group without E-cad-Fc. By contrast, a longer exposure to the same chimeric protein resulted in higher proportions of trophoblastic vesicles. Thus, we establish an important role of E-cadherin-mediated adherens junctions in promoting both the division of single 1/8 blastomeres and the efficiency of the ESC derivation process. [ABSTRACT FROM AUTHOR]

Purpose: The aim of the present study is to compare three previously described mouse embryonic stem cell derivation methods to evaluate the influence of culture conditions, number of isolated blastomeres and embryonic stage in the derivation process. Methods: Three embryonic stem cell derivation methods: standard, pre-adhesion and defined culture medium method, were compared in the derivation from isolated blastomeres and whole embryos at 4- and 8-cell stages. Results: A total of 200 embryonic stem cell lines were obtained with an efficiency ranging from 1.9% to 72%. Conclusions: Using either isolated blastomeres or whole embryos, the highest rates of mouse embryonic stem cell establishment were achieved with the defined culture medium method and efficiencies increased as development progressed. Using isolated blastomeres, efficiencies increased in parallel to the proportion of the embryo volume used to start the derivation process. [ABSTRACT FROM AUTHOR]

Cell handling is currently hindered by rudimentary-manufactured manipulators. Restrictive designs of glass pipettes and other micromanipulators limit functionality and often damage cells, ultimately resulting in lysis. We present a novel technique to design and mill conventional glass pipettes at specifically chosen angles and geometries. Focus ion beam milling by Ga+ ions yields extremely polished edges. Results from mouse embryo piercing correlate increased penetration rates with decreased pipette angle. Milled pipettes maintain structural integrity after repeated piercing. For the first time, the effects of unintentionally implanted Ga+ on embryo development are addressed. Optimum embryo development up to blastocyst stage after manipulation reveal little impact of residual implanted Ga+, suggesting biocompatibility and paving the way to introducing ion milling techniques in the biomedical device arena. The milling technique can be adequately tailored to specific applications and allows for mass production, presenting a promising avenue for future, increasingly demanding, cell handling. [ABSTRACT FROM AUTHOR]

Quality of life (QoL) is a significant parameter in estimating the social and interpersonal impact of psychopathology, clinical course, and response to treatment in many mental illnesses, particularly schizophrenia. The clinician’s assessment and patient’s subjective report are essential ingredients of this process. We describe the development and structure of a QoL questionnaire based on the patient’s subjective understanding of the problems. Ninety areas identified by four groups of experts resulted in a 126-item questionnaire administered to 279 patients with a DSM-IV diagnosis of schizophrenia. A factorial approach led to 59 total items and two final scales (favorable and unfavorable), with three factors each. The Seville QoL Questionnaire (SQLQ) results can be presented as individual scores for each scale, or a total for both. Its psychometric properties have proven quite acceptable in reliability and validity, comparing favorably with other scales. SQLQ may prove to be a useful addition to QoL-measuring instruments. [ABSTRACT FROM AUTHOR]

Three different parameters (temperature, solvent, and extraction time) were studied regarding to pressure liquid extraction (PLE) of antioxidant and antimicrobial compounds from Spirulina platensis. Two different antioxidant methods, β-carotene bleaching method and DPPH• (2,2-Diphenyl-1-picrylhydrazyl hydrate) free radical scavenging assay, were used to determine the optimal PLE conditions for antioxidants extraction. The selected conditions were as follows: extraction temperature equal to 115 °C, extraction time equal to 15 min and ethanol as extracting solvent. The main antioxidant compounds found in this extract were identified as zeaxanthin, a myxoxanthophyll-like compound and very polar phenolic compounds. Moreover, antimicrobial activity of different PLE fractions was tested against Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 11775, Candida albicans ATCC 60193, and Aspergillus niger ATCC 16404. Data obtained showed the hexane and petroleum ether extracts were slightly more active than ethanolic extracts. As for water extracts, none of them were active against the microorganisms tested. Data indicated that both 115 and 170 °C were the best extraction temperatures conditions in order to optimize the extraction of antimicrobial compounds, whereas 9 min was the optimal extraction time. Besides, C. albicans was the most sensitive microorganism to all Spirulina PLE extracts. [ABSTRACT FROM AUTHOR]

Therapeutic drug carriers can drive their cargo to their target cells. However, an obstacle is usually the entrapment of the drug inside the endolysosomal compartment, which physically impedes its actuation by the impossibility of reaching its molecular site of action. To overcome this hurdle, photochemical internalization (PCI) has been proposed, but the extent of PCI-induced membrane disruption and its capability to allow the release of microparticles is unknown. The aim of the present study was to determine if PCI allows the release of microparticles from the endolysosomal compartment to the cytosol and to analyze at the ultrastructural level the effect of PCI on the membrane surrounding the particles. Confocal microscope allowed us to detect that endolysosomal membranes suffered some disruption after PCI, evidenced by the diffusion of soluble transferrin from the endolysosomes to the cytosol and by a decrease of LAMP1-microparticles co-localization. Transmission electron microscopy (TEM) showed a decrease in the number of well-defined membranes around microparticles after PCI, and scanning TEM combined with energy dispersive x-ray revealed an increase in the width of endolysosomal membranes after treatment. These results suggest that endolysosomal membranes suffered an ultrastructure alteration after PCI, enough to liberate soluble transferrin but not the entire microparticles. [ABSTRACT FROM AUTHOR]

The use of micro- and nanodevices as multifunctional systems for biomedical applications has experienced an exponential growth during the past decades. Although a large number of studies have focused on the design and fabrication of new micro- and nanosystems capable of developing multiple functions, a deeper understanding of their interaction with cells is required. In the present study, we evaluated the effect of different microparticle surfaces on their interaction with normal and tumoral human breast epithelial cell lines. For this, AlexaFluor488 IgG functionalized polystyrene microparticles (3 μm) were coated with Polyethyleneimine (PEI) at two different molecular weights, 25 and 750 kDa. The effect of microparticle surface properties on cytotoxicity, cellular uptake and endocytic pathways were assessed for both normal and tumoral cell lines. Results showed a differential response between the two cell lines regarding uptake efficiency and mechanisms of endocytosis, highlighting the potential role of microparticle surface tunning for specific cell targeting. [ABSTRACT FROM AUTHOR]