Insulin induces PFKFB3 gene expression in HT29 human colon adenocarcinoma cells

AbstractFructose 2,6-bisphosphate is present at high concentrations in many established lines of transformed cells. It plays a key role in the maintenance of a high glycolytic rate by coupling hormonal and growth factor signals with metabolic demand. The concentration of fructose 2,6-bisphosphate is controlled by the activity of the homodimeric bifunctional enzyme 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFK-2). We report here the PFKFB-3 gene expression control by insulin in the human colon adenocarcinoma HT29 cell line. The incubation of these cells with 1 μM insulin resulted in an increase in the PFK-2 mRNA level after 6 h of treatment, this effect being blocked by actinomycin D. Furthermore, insulin induced ubiquitous PFK-2 protein levels, that were evident after a lag of 3 h and could be inhibited by incubation with cycloheximide

Higher codimension braneworlds from intersecting branes

We study the matching conditions of intersecting brane worlds in Lovelock gravity in arbitrary dimension. We show that intersecting various codimension 1 and/or codimension 2 branes one can find solutions that represent energy-momentum densities localized in the intersection, providing thus the first examples of infinitesimally thin higher codimension braneworlds that are free of singularities and where the backreaction of the brane in the background is fully taken into account.Comment: 20 pages; v2. references and comments added to match the published versio

Design of a case management model for people with chronic disease (Heart Failure and COPD). Phase I: modeling and identification of the main components of the intervention through their actors: patients and professionals (DELTA-ICE-PRO Study

Background Chronic diseases account for nearly 60% of deaths around the world. The extent of this silent epidemic has not met determined responses in governments, policies or professionals in order to transform old Health Care Systems, configured for acute diseases. There is a large list of research about alternative models for people with chronic conditions, many of them with an advanced practice nurse as a key provider, as case management. But some methodological concerns raise, above all, the design of the intervention (intensity, frequency, components, etc). Methods/Design Objectives: General: To develop the first and second phases (theorization and modeling) for designing a multifaceted case-management intervention in people with chronic conditions (COPD and heart failure) and their caregivers. Specific aims: 1) To identify key events in people living with chronic disease and their relation with the Health Care System, from their point of view. 2) To know the coping mechanisms developed by patients and their caregivers along the story with the disease. 3) To know the information processing and its utilization in their interactions with health care providers. 4) To detect potential unmet needs and the ways deployed by patients and their caregivers to resolve them. 5) To obtain a description from patients and caregivers, about their itineraries along the Health Care System, in terms of continuity, accessibility and comprehensiveness of care. 6) To build up a list of promising case-management interventions in patients with Heart Failure and COPD with this information in order to frame it into theoretical models for its reproducibility and conceptualization. 7) To undergo this list to expert judgment to assess its feasibility and pertinence in the Andalusian Health Care. Design: Qualitative research with two phases: For the first five objectives, a qualitative technique with biographic stories will be developed and, for the remaining objectives, an expert consensus through Delphi technique, on the possible interventions yielded from the first phase. The study will be developed in the provinces of Almería, Málaga and Granada in the Southern Spain, from patients included in the Andalusian Health Care Service database with the diagnosis of COPD or Heart Failure, with the collaboration of case manager nurses and general practitioners for the assessment of their suitability to inclusion criteria. Patients and caregivers will be interviewed in their homes or their Health Centers, with their family or their case manager nurse as mediator. Discussion First of a series of studies intended to design a case-management service for people with heart failure and COPD, in the Andalusian Health Care System, where case management has been implemented since 2002. Accordingly with the steps of a theoretical model for complex interventions, in this study, theorization and intervention modeling phases will be developed.This research was carried out with the support of one research grant, awarded by the Regional Health Ministry of Andalusia (Exp. 0222/2008

Einstein-Planck Formula, Equivalence Principle, and Black Hole Radiance

The presence of gravity implies corrections to the Einstein-Planck formula E = hν. This gives hope that the divergent blueshift in frequency, associated to the presence of a black hole horizon, could be smoothed out for the energy. Using simple arguments based on Einstein's equivalence principle, we show that this is only possible if a black hole emits, in a first approximation, not just a single particle, but thermal radiation

Cucurbit chlorotic yellows virus p22 suppressor of RNA silencing binds single-, double-stranded long and short interfering RNA molecules in vitro

[EN] Cucurbit chlorotic yellows virus (CCYV) is a new member of the genus Crinivirus (family Closteroviridae) with a bi-partite genome. CCYV RNA 1-encoded p22 has recently been reported to be a weak local suppressor of RNA silencing for which an interaction with cucumber SKP1LB1 through an F-box-like motif was demonstrated to be essential. Using a bacterially expressed maltose-binding protein (MBP) fusion of CCYV p22 in electrophoretic mobility shift assays (EMSA), we have examined in vitro its ability to bind different RNA templates. Our experiments showed that CCYV p22 is able to bind to ss and ds long RNAs, in addition to ss and ds small interfering (si) RNA molecules. CCYV p22 deletion mutants (MBP_CCYV DEL1-4) were produced that covered the entire protein, with MBP_CCYV DEL2 corresponding to the F-box motif and its flanking sequences. None of these deletions abolished the capacity of CCYV p22 to bind ss- and dsRNA molecules. However, deletions affecting the C-terminal half of the protein resulted in decreased binding efficiency for either ss- or dsRNA molecules indicating that essential elements for these interactions are located in this region. Taken together, our data add to current knowledge of the mode of action of suppressors of RNA silencing encoded by genes sited at the 3'-terminus of crinivirus genomic RNA 1, and shed light on the involvement of CCYV p22 in the suppression of RNA silencing and/or in another role in the virus life cycle via RNA binding.Salavert, F.; Navarro Bohigues, JA.; Owen, CA.; Khechmar, S.; Pallás Benet, V.; Livieratos, IC. (2020). Cucurbit chlorotic yellows virus p22 suppressor of RNA silencing binds single-, double-stranded long and short interfering RNA molecules in vitro. Virus Research. 279:1-8. https://doi.org/10.1016/j.virusres.2020.197887S18279Abrahamian, P. E., Seblani, R., Sobh, H., & Abou-Jawdah, Y. (2013). Detection and quantitation of two cucurbit criniviruses in mixed infection by real-time RT-PCR. Journal of Virological Methods, 193(2), 320-326. doi:10.1016/j.jviromet.2013.06.004Bananej, K., Menzel, W., Kianfar, N., Vahdat, A., & Winter, S. (2013). First Report of Cucurbit chlorotic yellows virus Infecting Cucumber, Melon, and Squash in Iran. Plant Disease, 97(7), 1005-1005. doi:10.1094/pdis-01-13-0125-pdnBrantley, J. D., & Hunt, A. G. (1993). The N-terminal protein of the polyprotein encoded by the potyvirus tobacco vein mottling virus is an RNA-binding protein. Journal of General Virology, 74(6), 1157-1162. doi:10.1099/0022-1317-74-6-1157BUCK, K. W., & KEMPSON-JONES, G. F. (1970). Three Types of Virus Particle in Penicillium stoloniferum. Nature, 225(5236), 945-946. doi:10.1038/225945a0Cañizares, M. C., Navas-Castillo, J., & Moriones, E. (2008). Multiple suppressors of RNA silencing encoded by both genomic RNAs of the crinivirus, Tomato chlorosis virus. Virology, 379(1), 168-174. doi:10.1016/j.virol.2008.06.020Chen, S., Sun, X., Shi, Y., Wei, Y., Han, X., Li, H., … Shi, Y. (2019). Cucurbit Chlorotic Yellows Virus p22 Protein Interacts with Cucumber SKP1LB1 and Its F-Box-Like Motif Is Crucial for Silencing Suppressor Activity. Viruses, 11(9), 818. doi:10.3390/v11090818Cuellar, W. J., Tairo, F., Kreuze, J. F., & Valkonen, J. P. T. (2008). Analysis of gene content in sweet potato chlorotic stunt virus RNA1 reveals the presence of the p22 RNA silencing suppressor in only a few isolates: implications for viral evolution and synergism. Journal of General Virology, 89(2), 573-582. doi:10.1099/vir.0.83471-0Cuellar, W. J., Kreuze, J. F., Rajamaki, M.-L., Cruzado, K. R., Untiveros, M., & Valkonen, J. P. T. (2009). Elimination of antiviral defense by viral RNase III. Proceedings of the National Academy of Sciences, 106(25), 10354-10358. doi:10.1073/pnas.0806042106GYOUTOKU, Y., OKAZAKI, S., FURUTA, A., ETOH, T., MIZOBE, M., KUNO, K., … OKUDA, M. (2009). Chlorotic yellows disease of melon caused by Cucurbit chlorotic yellows virus, a new crinivirus. Japanese Journal of Phytopathology, 75(2), 109-111. doi:10.3186/jjphytopath.75.109Herranz, M. C., & Pallás, V. (2004). RNA-binding properties and mapping of the RNA-binding domain from the movement protein of Prunus necrotic ringspot virus. Journal of General Virology, 85(3), 761-768. doi:10.1099/vir.0.19534-0Huang, L.-H., Tseng, H.-H., Li, J.-T., & Chen, T.-C. (2010). First Report of Cucurbit chlorotic yellows virus Infecting Cucurbits in Taiwan. Plant Disease, 94(9), 1168-1168. doi:10.1094/pdis-94-9-1168bKataya, A. R. A., Suliman, M. N. S., Kalantidis, K., & Livieratos, I. C. (2009). Cucurbit yellow stunting disorder virus p25 is a suppressor of post-transcriptional gene silencing. Virus Research, 145(1), 48-53. doi:10.1016/j.virusres.2009.06.010Klaassen, V. A., Mayhew, D., Fisher, D., & Falk, B. W. (1996). In VitroTranscripts from Cloned cDNAs of the Lettuce Infectious Yellows Closterovirus Bipartite Genomic RNAs Are Competent for Replication inNicotiana benthamianaProtoplasts. Virology, 222(1), 169-175. doi:10.1006/viro.1996.0407Kreuze, J. F., Savenkov, E. I., Cuellar, W., Li, X., & Valkonen, J. P. T. (2005). Viral Class 1 RNase III Involved in Suppression of RNA Silencing. Journal of Virology, 79(11), 7227-7238. doi:10.1128/jvi.79.11.7227-7238.2005Kubota, K., & Ng, J. C. K. (2016). Lettuce chlorosis virus P23 Suppresses RNA Silencing and Induces Local Necrosis with Increased Severity at Raised Temperatures. Phytopathology®, 106(6), 653-662. doi:10.1094/phyto-09-15-0219-rLandeo-Ríos, Y., Navas-Castillo, J., Moriones, E., & Cañizares, M. C. (2016). The p22 RNA silencing suppressor of the crinivirus Tomato chlorosis virus preferentially binds long dsRNAs preventing them from cleavage. Virology, 488, 129-136. doi:10.1016/j.virol.2015.11.008Marcos, J. F., Vilar, M., Pérez-Payá, E., & Pallás, V. (1999). In VivoDetection, RNA-Binding Properties and Characterization of the RNA-Binding Domain of the p7 Putative Movement Protein from Carnation Mottle Carmovirus (CarMV). Virology, 255(2), 354-365. doi:10.1006/viro.1998.9596Mashiko, T., Wang, W.-Q., Hartono, S., Suastica, G., Neriya, Y., Nishigawa, H., & Natsuaki, T. (2019). The p27 open reading frame of tomato infectious chlorosis virus encodes a suppressor of RNA silencing. Journal of General Plant Pathology, 85(4), 301-305. doi:10.1007/s10327-019-00850-0Navarro, J. A., Genovés, A., Climent, J., Saurí, A., Martínez-Gil, L., Mingarro, I., & Pallás, V. (2006). RNA-binding properties and membrane insertion of Melon necrotic spot virus (MNSV) double gene block movement proteins. Virology, 356(1-2), 57-67. doi:10.1016/j.virol.2006.07.040Okuda, M., Okazaki, S., Yamasaki, S., Okuda, S., & Sugiyama, M. (2010). Host Range and Complete Genome Sequence of Cucurbit chlorotic yellows virus, a New Member of the Genus Crinivirus. Phytopathology®, 100(6), 560-566. doi:10.1094/phyto-100-6-0560Orfanidou, C., Maliogka, V. I., & Katis, N. I. (2014). First Report of Cucurbit chlorotic yellows virus in Cucumber, Melon, and Watermelon in Greece. Plant Disease, 98(10), 1446-1446. doi:10.1094/pdis-03-14-0311-pdnOrfanidou, C. G., Mathioudakis, M. M., Katsarou, K., Livieratos, I., Katis, N., & Maliogka, V. I. (2019). Cucurbit chlorotic yellows virus p22 is a suppressor of local RNA silencing. Archives of Virology, 164(11), 2747-2759. doi:10.1007/s00705-019-04391-xOrílio, A. F., Fortes, I. M., & Navas-Castillo, J. (2014). Infectious cDNA clones of the crinivirus Tomato chlorosis virus are competent for systemic plant infection and whitefly-transmission. Virology, 464-465, 365-374. doi:10.1016/j.virol.2014.07.032Pumplin, N., & Voinnet, O. (2013). RNA silencing suppression by plant pathogens: defence, counter-defence and counter-counter-defence. Nature Reviews Microbiology, 11(11), 745-760. doi:10.1038/nrmicro3120Richmond, K. E., Chenault, K., Sherwood, J. L., & German, T. L. (1998). Characterization of the Nucleic Acid Binding Properties of Tomato Spotted Wilt Virus Nucleocapsid Protein. Virology, 248(1), 6-11. doi:10.1006/viro.1998.9223Salem, N. M., Chen, A. Y. S., Tzanetakis, I. E., Mongkolsiriwattana, C., & Ng, J. C. K. (2009). Further complexity of the genus Crinivirus revealed by the complete genome sequence of Lettuce chlorosis virus (LCV) and the similar temporal accumulation of LCV genomic RNAs 1 and 2. Virology, 390(1), 45-55. doi:10.1016/j.virol.2009.04.025Serra-Soriano, M., Antonio Navarro, J., & Pallás, V. (2016). Dissecting the multifunctional role of the N-terminal domain of theMelon necrotic spot viruscoat protein in RNA packaging, viral movement and interference with antiviral plant defence. Molecular Plant Pathology, 18(6), 837-849. doi:10.1111/mpp.12448Simon, A. E., & Miller, W. A. (2013). 3′ Cap-Independent Translation Enhancers of Plant Viruses. Annual Review of Microbiology, 67(1), 21-42. doi:10.1146/annurev-micro-092412-155609Voinnet, O. (2005). Induction and suppression of RNA silencing: insights from viral infections. Nature Reviews Genetics, 6(3), 206-220. doi:10.1038/nrg1555Wang, J., Stewart, L. R., Kiss, Z., & Falk, B. W. (2010). Lettuce infectious yellows virus (LIYV) RNA 1-encoded P34 is an RNA-binding protein and exhibits perinuclear localization. Virology, 403(1), 67-77. doi:10.1016/j.virol.2010.04.006Weinheimer, I., Boonrod, K., Moser, M., Wassenegger, M., Krczal, G., Butcher, S. J., & Valkonen, J. P. T. (2014). Binding and processing of small dsRNA molecules by the class 1 RNase III protein encoded by sweet potato chlorotic stunt virus. Journal of General Virology, 95(2), 486-495. doi:10.1099/vir.0.058693-0Yeh, H.-H., Tian, T., Rubio, L., Crawford, B., & Falk, B. W. (2000). Asynchronous Accumulation of Lettuce Infectious Yellows Virus RNAs 1 and 2 and Identification of an RNA 1 trans Enhancer of RNA 2 Accumulation. Journal of Virology, 74(13), 5762-5768. doi:10.1128/jvi.74.13.5762-5768.200

Leadership, management, quality, and innovation in organ donation: 2019 Kunming recommendations for One Belt & One Road countries

The findings and recommendations of the 2019 consensus conference in organ donation, held in Kunming, China, are here reported. The main objective of the conference was to gather relevant information from experts involved in the field. The data and opinions provided allowed to propose a series of recommendations for “One Belt & One Road Countries” on how to achieve self-sufficiency in organ donation. Leadership in organ donation should be results-oriented and goal-driven based on the principles of excellence, empowerment, and engagement, providing the means, resources, and strategies necessary to reach the goal in earnest. Management includes good governance and transparency of a national registry of patients in the waiting list, donors, transplants, transplant teams, quality, and safety programs with continuous educational training of health care professionals. Mandatory monitoring, auditing and evaluation of quality must be incorporated into donation practices as relevant points in innovation, as well as the adoption of already established and novel processes and technologies. Achievement of self-sufficiency in organ donation is a crucial step to fight against transplant tourism and to prevent organ trafficking. Based on recommendations arising from the conference, each country could review and develop individualized action plans adjusted to its own circumstances and reality

Recapitulating the Bayesian framework for neurosurgical outpatient care and a cost-benefit analysis of telemedicine for socioeconomically disadvantaged patients in the Philippines during the pandemic

Objective: In the Philippines during recent months, a neurosurgical center that caters primarily to socioeconomically disadvantaged patients has encountered unprecedented changes in practice patterns brought about by the COVID-19 pandemic. In particular, the usual task of outpatient care has shifted to the telemedicine format, bringing along all of its attendant advantages and gargantuan challenges. The authors sought to determine the responsiveness of this telemedicine setup to the needs of their disadvantaged patients and explored the application of Bayesian inference to enhance the use of teleconsultation in daily clinical decision-making. Methods: The authors used the following methods to assess the telemedicine setup used in a low-resource setting during the pandemic: 1) a cross-sectional survey of patients who participated in a medical consultation via telemedicine during the 16-week period from March 16, 2020, to July 15, 2020; 2) a cost-benefit analysis of the use of telemedicine by patients; and 3) a case illustration of a Bayesian approach application unique to the teleconsultation scenario. Results: Of the 272 patient beneficiaries of telemedicine in a 16-week period, 57 responded to the survey. The survey responses regarding neurosurgical outpatient care through telemedicine yielded high ratings of utility for the patients and their caregivers. According to 64% of respondents, the affordability of the telemedicine setup also prevented them from borrowing money from others, among other adverse life events prevented. There were realized financial gains on the part of the patients in terms of cost savings and protection from further impoverishment. The benefit-cost ratio was 3.51 for the patients, signifying that the benefits outweighed the costs. An actual teleconsultation case vignette was reported that is meant to be instructive and contributory to the preparedness of the neurosurgeon on the provider end of the service delivery. Conclusions: Telemedicine holds promise as a viable and safe method for health service delivery during the pandemic. In the setting of a health system that is continually challenged by shortages of resources, this study shows that an effective telemedicine setup can come with high benefit-cost ratios and quality of care, along with the assurance of patient satisfaction. The potential for high-quality care can be enhanced by the inclusion of the Bayesian framework to the basic toolkit of remote clinical assessment. When confronted with choices in terms of differential diagnosis and tests, the rigor of a simple application of the Bayesian framework can minimize costs arising from uncertaintie