Non-lethal PCR genotyping of single Drosophila

In Drosophila, genetic techniques relying on stochastic chromosomal rearrangements involve the generation and screening of a large number of fly stocks to isolate a few lines of interest. Here, we describe a PCR-based method allowing non-lethal molecular characterization of single flies. Using this procedure, individual candidate recombinant animals can be genotyped and selected one generation earlier than with extant methodology and, importantly, before stocks are established. This advance should significantly facilitate several of the most fundamental and routine techniques in Drosophila genetics

Virtual rehabilitation for multiple sclerosis using a Kinect-based system: randomized controlled trial

©Jose-Antonio Lozano-Quilis, Hermenegildo Gil-Gómez, Jose-Antonio Gil-Gómez, Sergio Albiol-Pérez, Guillermo Palacios-Navarro, Habib M Fardoun, Abdulfattah S Mashat. Originally published in JMIR Serious Games (http://games.jmir.org), 12.11.2014. This is an open-access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work, first published in JMIR Serious Games, is properly cited. The complete bibliographic information, a link to the original publication on http://games.jmir.org, as well as this copyright and license information must be included.Background: The methods used for the motor rehabilitation of patients with neurological disorders include a number of different rehabilitation exercises. For patients who have been diagnosed with multiple sclerosis (MS), the performance of motor rehabilitation exercises is essential. Nevertheless, this rehabilitation may be tedious, negatively influencing patients motivation and adherence to treatment. Objective: We present RemoviEM, a system based on Kinect that uses virtual reality (VR) and natural user interfaces (NUI) to offer patients with MS an intuitive and motivating way to perform several motor rehabilitation exercises. It offers therapists a new motor rehabilitation tool for the rehabilitation process, providing feedback on the patient s progress. Moreover, it is a low-cost system, a feature that can facilitate its integration in clinical rehabilitation centers. Methods: A randomized and controlled single blinded study was carried out to assess the influence of a Kinect-based virtual rehabilitation system on the balance rehabilitation of patients with MS. This study describes RemoviEM and evaluates its effectiveness compared to standard rehabilitation. To achieve this objective, a clinical trial was carried out. Eleven patients from a MS association participated in the clinical trial. The mean age was 44.82 (SD 10.44) and the mean time from diagnosis (years) was 9.77 (SD 10.40). Clinical effectiveness was evaluated using clinical balance scales. Results: Significant group-by-time interaction was detected in the scores of the Berg Balance Scale (P=.011) and the Anterior Reach Test in standing position (P=.011). Post-hoc analysis showed greater improvement in the experimental group for these variables than in the control group for these variables. The Suitability Evaluation Questionnaire (SEQ) showed good results in usability, acceptance, security, and safety for the evaluated system. Conclusions: The results obtained suggest that RemoviEM represents a motivational and effective alternative to traditional motor rehabilitation for MS patients. These results have encouraged us to improve the system with new exercises, which are currently being developed.This contribution was partially funded by the Generalitat Valenciana ("Ajudes per a la realitzacio de projectes d'I+D per a grups d'investigacion emergents", projecte GV/2012/069) and by the Fundacion Antonio Gargallo ("Ayudas financiadas por la Obra Social de Ibercaja de proyectos de investigacion 2013", proyecto 2013/B001).Lozano Quilis, JA.; Gil Gómez, H.; Gil-Gómez, J.; Albiol Pérez, S.; Palacios Navarro, G.; Fardoun, HM.; Mashat, AS. (2014). Virtual rehabilitation for multiple sclerosis using a Kinect-based system: randomized controlled trial. JMIR Serious Games. 2(2). https://doi.org/10.2196/games.2933Se1222Adamovich, S. V., Fluet, G. G., Tunik, E., & Merians, A. S. (2009). Sensorimotor training in virtual reality: A review. Neurorehabilitation, 25(1), 29-44. doi:10.3233/nre-2009-0497Lange, B., Flynn, S., Proffitt, R., Chang, C.-Y., & «Skip» Rizzo, A. (2010). Development of an Interactive Game-Based Rehabilitation Tool for Dynamic Balance Training. Topics in Stroke Rehabilitation, 17(5), 345-352. doi:10.1310/tsr1705-345Lozano, J. A., Montesa, J., Juan, M. C., Alcañiz, M., Rey, B., Gil, J., … Morganti, F. (2005). VR-Mirror: A Virtual Reality System for Mental Practice in Post-Stroke Rehabilitation. Lecture Notes in Computer Science, 241-251. doi:10.1007/11536482_23Lange, B. S., Requejo, P., Flynn, S. M., Rizzo, A. A., Valero-Cuevas, F. J., Baker, L., & Winstein, C. (2010). The Potential of Virtual Reality and Gaming to Assist Successful Aging with Disability. Physical Medicine and Rehabilitation Clinics of North America, 21(2), 339-356. doi:10.1016/j.pmr.2009.12.007Wuang, Y.-P., Chiang, C.-S., Su, C.-Y., & Wang, C.-C. (2011). Effectiveness of virtual reality using Wii gaming technology in children with Down syndrome. Research in Developmental Disabilities, 32(1), 312-321. doi:10.1016/j.ridd.2010.10.002Chang, Y.-J., Chen, S.-F., & Huang, J.-D. (2011). A Kinect-based system for physical rehabilitation: A pilot study for young adults with motor disabilities. Research in Developmental Disabilities, 32(6), 2566-2570. doi:10.1016/j.ridd.2011.07.002Da GamaAChavezTFigueiredoLTeichriebVPoster: improving motor rehabilitation process through a natural interaction based system using kinect sensor, IEEE Symposium on 3D User Interfaces 2012: 145-1462012IEEE Symposium on 3D User InterfacesMar 4-5Costa Mesa, CABaram, Y., & Miller, A. (2006). Virtual reality cues for improvement of gait in patients with multiple sclerosis. Neurology, 66(2), 178-181. doi:10.1212/01.wnl.0000194255.82542.6bFulk, G. D. (2005). Locomotor Training and Virtual Reality-based Balance Training for an Individual with Multiple Sclerosis. Journal of Neurologic Physical Therapy, 29(1), 34-42. doi:10.1097/01.npt.0000282260.59078.e4Lozano-QuilisJAAlbiol-PerezSGil-GomezHPalaciosGFardoumHMGil-GomezJAMashatASVirtual reality system for multiple sclerosis rehabilitation using KINECT2013International Conference on Pervasive Computing Technologies for Healthcare (PervasiveHealth)May 5-8Venice, Italy366369Van Hedel, H. J., Wirz, M., & Dietz, V. (2005). Assessing walking ability in subjects with spinal cord injury: Validity and reliability of 3 walking tests. Archives of Physical Medicine and Rehabilitation, 86(2), 190-196. doi:10.1016/j.apmr.2004.02.010Podsiadlo, D., & Richardson, S. (1991). The Timed «Up & Go»: A Test of Basic Functional Mobility for Frail Elderly Persons. Journal of the American Geriatrics Society, 39(2), 142-148. doi:10.1111/j.1532-5415.1991.tb01616.xSteffen, T. M., Hacker, T. A., & Mollinger, L. (2002). Age- and Gender-Related Test Performance in Community-Dwelling Elderly People: Six-Minute Walk Test, Berg Balance Scale, Timed Up & Go Test, and Gait Speeds. Physical Therapy, 82(2), 128-137. doi:10.1093/ptj/82.2.128Gil-GomezJASEQ: Suitability Evaluation Questionnaire for Virtual Rehabilitation systems2013International Conference on Pervasive Computing Technologies for Healthcare (PervasiveHealth)2013Venice, Italy33533

Heterogeneity of Skin Surface Oxygen Level of Wrist in Relation to Acupuncture Point

The distribution of partial oxygen pressure (pO2) is analyzed for the anterior aspect of the left wrist with an amperometric oxygen microsensor composed of a small planar Pt disk-sensing area (diameter = 25 μm). The pO2 levels vary depending on the measurement location over the wrist skin, and they are systematically monitored in the analysis for both one-dimensional single line (along the wrist transverse crease) and two-dimensional square area of the wrist region. Relatively higher pO2 values are observed at certain area in close proximity to the position of acupuncture points with statistical significance, indicating strong relationship between oxygen and acupuncture point. The used oxygen microsensor is sensitive enough to detect the pO2 variation depending on the location. This study may provide information helpful to understand possible physiological roles of the acupuncture points

Phosphorescent sensor for biological mobile zinc

A new phosphorescent zinc sensor (ZIrF) was constructed, based on an Ir(III) complex bearing two 2-(2,4-difluorophenyl)pyridine (dfppy) cyclometalating ligands and a neutral 1,10-phenanthroline (phen) ligand. A zinc-specific di(2-picolyl)amine (DPA) receptor was introduced at the 4-position of the phen ligand via a methylene linker. The cationic Ir(III) complex exhibited dual phosphorescence bands in CH[subscript 3]CN solutions originating from blue and yellow emission of the dfppy and phen ligands, respectively. Zinc coordination selectively enhanced the latter, affording a phosphorescence ratiometric response. Electrochemical techniques, quantum chemical calculations, and steady-state and femtosecond spectroscopy were employed to establish a photophysical mechanism for this phosphorescence response. The studies revealed that zinc coordination perturbs nonemissive processes of photoinduced electron transfer and intraligand charge-transfer transition occurring between DPA and phen. ZIrF can detect zinc ions in a reversible and selective manner in buffered solution (pH 7.0, 25 mM PIPES) with K[subscript d] = 11 nM and pK[subscript a] = 4.16. Enhanced signal-to-noise ratios were achieved by time-gated acquisition of long-lived phosphorescence signals. The sensor was applied to image biological free zinc ions in live A549 cells by confocal laser scanning microscopy. A fluorescence lifetime imaging microscope detected an increase in photoluminescence lifetime for zinc-treated A549 cells as compared to controls. ZIrF is the first successful phosphorescent sensor that detects zinc ions in biological samples.National Institute of General Medical Sciences (U.S.) (Grant GM065519)Ewha Woman's University (Korea) (RP-Grant 2010

PKCε-mediated ERK1/2 activation involved in radiation-induced cell death in NIH3T3 cells

AbstractProtein kinase C (PKC) isoforms play distinct roles in cellular functions. We have previously shown that ionizing radiation activates PKC isoforms (α, δ, ε, and ζ), however, isoform-specific sensitivities to radiation and its exact mechanisms in radiation mediated signal transduction are not fully understood. In this study, we showed that overexpression of PKC isoforms (α, δ, ε, and ζ) increased radiation-induced cell death in NIH3T3 cells and PKCε overexpression was predominantly responsible. In addition, PKCε overexpression increased ERK1/2 activation without altering other MAP-kinases such as p38 MAPK or JNK. Co-transfection of dominant negative PKCε (PKCε-KR) blocked both PKCε-mediated ERK1/2 activation and radiation-induced cell death, while catalytically active PKCε construction augmented these phenomena. When the PKCε overexpressed cells were pretreated with PD98059, MEK inhibitor, radiation-induced cell death was inhibited. Co-transfection of the cells with a mutant of ERK1 or -2 (ERK1-KR or ERK2-KR) also blocked these phenomena, and co-transfection with dominant negative Ras or Raf cDNA revealed that PKCε-mediated ERK1/2 activation was Ras–Raf-dependent. In conclusion, PKCε-mediated ERK1/2 activation was responsible for the radiation-induced cell death

Heterometallic Titanium-Organic Frameworks as Dual Metal Catalysts for Synergistic Non-Buffered Hydrolysis of Nerve Agent Simulants

Heterometallic metal-organic frameworks (MOFs) can offer important advantages over their homometallic counterparts to enable targeted modification of their adsorption, structural response, electronic structure, or chemical reactivity. However, controlling metal distribution in these solids still remains a challenge. The family of mesoporous titanium-organic frameworks, MUV-101(M), displays heterometallic TiM2 nodes assembled from direct reaction of Ti(IV) and M(II) salts. We use the degradation of nerve agent simulants to demonstrate that only TiFe2 nodes are capable of catalytic degradation in non-buffered conditions. By using an integrative experimental-computational approach, we rationalize how the two metals influence each other, in this case, for a synergistic mechanism reminiscent of bimetallic enzymes. Our results highlight the importance of controlling metal distribution at an atomic level to span the interest of heterometallic MOFs to a broad scope of cascade or tandem reactions. Summary Mixed-metal or heterometallic metal-organic frameworks (MOFs) are gaining importance as a route to produce materials with increasing chemical and functional complexities. We report a family of heterometallic titanium frameworks, MUV-101(M), and use them to exemplify the advantages of controlling metal distribution across the framework in heterogeneous catalysis by exploring their activity toward the degradation of a nerve agent simulant of Sarin gas. MUV-101(Fe) is the only pristine MOF capable of catalytic degradation of diisopropyl-fluorophosphate (DIFP) in non-buffered aqueous media. This activity cannot be explained only by the association of two metals, but to their synergistic cooperation, to create a whole that is more efficient than the simple sum of its parts. Our simulations suggest a dual-metal mechanism reminiscent of bimetallic enzymes, where the combination of Ti(IV) Lewis acid and Fe(III)–OH Brönsted base sites leads to a lower energy barrier for more efficient degradation of DIFP in absence of a base.Financial support for this work was provided by the Marie Skłodowska-Curie Global Fellowships (749359-EnanSET, N.M.P) within the European Union research and innovation framework programme (2014-2020

Under-five mortality: spatial-temporal clusters in Ifakara HDSS in South-eastern Tanzania.

BACKGROUND\ud \ud Childhood mortality remains an important subject, particularly in sub-Saharan Africa where levels are still unacceptably high. To achieve the set Millennium Development Goals 4, calls for comprehensive application of the proven cost-effective interventions. Understanding spatial clustering of childhood mortality can provide a guide in targeting the interventions in a more strategic approach to the population where mortality is highest and the interventions are most likely to make an impact.\ud \ud METHODS\ud \ud Annual child mortality rates were calculated for each village, using person-years observed as the denominator. Kulldorff's spatial scan statistic was used for the identification and testing of childhood mortality clusters. All under-five deaths that occurred within a 10-year period from 1997 to 2006 were included in the analysis. Villages were used as units of clusters; all 25 health and demographic surveillance sites (HDSS) villages in the Ifakara health and demographic surveillance area were included.\ud \ud RESULTS\ud \ud Of the 10 years of analysis, statistically significant spatial clustering was identified in only 2 years (1998 and 2001). In 1998, the statistically significant cluster (p < 0.01) was composed of nine villages. A total of 106 childhood deaths were observed against an expected 77.3. The other statistically significant cluster (p < 0.05) identified in 2001 was composed of only one village. In this cluster, 36 childhood deaths were observed compared to 20.3 expected. Purely temporal analysis indicated that the year 2003 was a significant cluster (p < 0.05). Total deaths were 393 and expected were 335.8. Spatial-temporal analysis showed that nine villages were identified as statistically significant clusters (p < 0.05) for the period covering January 1997-December 1998. Total observed deaths in this cluster were 205 while 150.7 were expected.\ud \ud CONCLUSION\ud \ud There is evidence of spatial clustering in childhood mortality within the Ifakara HDSS. Further investigations are needed to explore the source of clustering and identify strategies of reaching the cluster population with the existing effective interventions. However, that should happen alongside delivery of interventions to the broader population