In silico single strand melting curve: a new approach to identify nucleic acid polymorphisms in Totiviridae

CpG-island promoters of developmental genes are unmethylated. DNA methylation state of CpG islands overlapping and surrounding the promoter region of Pax3 (a) and Pax7 (b) genes in myogenic (MB, MT, MF) and non-myogenic samples (ESC). CpG islands are indicated in green and regions analysed by sodium bisulphite sequencing are shown in red. Each circle represents a CpG dinucleotide and its distance to the gene TSS is indicated below. The colour gradient represents the percentage of methylation indicated in the legend. Abbreviations: ESC, embryonic stem cell; MB, myoblast; MT, myotube; MF, myofiber; TSS, transcription start site. c. DNA methylation state of -5 kb distal regulatory region for MyoD was analysed by sodium bisulphite sequencing in ESC and myoblast samples, and represented as above. (PDF 171 kb

Gestión por procesos y los indicadores de desempeño en la entidad pública distrital de Bambas, 2022

The study presented as a general objective of this study is to determine the relationship between process management and indicators in the performance of its district municipality of Bambas, 2022, for which it was based on the basic type, with a quantitative approach, to present numerical results. , with non-experimental design - Correlational and cross-sectional scope, working with a population of 20 workers and the sample was made up of the same amount, selected in a non-probabilistic way at the discretion of the researcher, applying the survey as a technique and the questionnaire as an instrument, carrying out validation by 3 experts and reliability by Cronbach's Alpha method, obtaining a value of 0.896 for the process management questionnaire and 0.880 for the performance indicators questionnaire, reaching the conclusion: Significant relationship was determined ( Rho = 0.940) between process management and indicators in the performance of its district municipality l de Bambas, 2022, showing that the actions carried out in process management are beneficial for it to be achieved in compliance with the performance indicators. Being demonstrated the general hypothesis of the studyEl estudio presento como objetivo general del presente estudio es determinar la relación entre la gestión de procesos y los indicadores en el desempeño de su municipalidad distrital de Bambas, 2022, para ello se basó en el tipo basica, con enfoque cuantitativo, por presentar resultados numéricos, con diseño no experimental – Correlacional y alcance transversal, trabajando con una población de 20 trabajadores y la muestra se conformó por la misma cantidad, seleccionada de manera no probabilística a criterio del investigador, aplicando como técnica la encuesta y como instrumento el cuestionario, realizando la validación por 3 expertos y la confiabilidad por el método de Alfa de Cronbach, obteniendo un valor de 0.896 para el cuestionario de la gestión por procesos y de 0.880 para el cuestionario de indicadores de desempeño, llegando a la conclusión: Se determinó relación significativa (Rho = 0.940) entre la gestión de procesos y los indicadores en el desempeño de su municipalidad distrital de Bambas, 2022, mostrando que las acciones que se realicen en la gestión por procesos son beneficiosas para que se logre en cumplimiento de los indicadores de desempeño. Quedando demostrada la hipótesis general del estudio

The E3 ubiquitin ligase SCF(Fbxo7) mediates proteasomal degradation of UXT isoform 2 (UXT-V2) to inhibit the NF-κB signaling pathway.

BACKGROUND: Ubiquitously eXpressed Transcript isoform 2 (UXTV2) is a prefoldin-like protein involved in NF-κB signaling, apoptosis, and the androgen and estrogen response. UXT-V2 is a cofactor in the NF-κB transcriptional enhanceosome, and its knockdown inhibits TNF-α -induced NF-κB activation. Fbxo7 is an F-box protein that interacts with SKP1, Cullin1 and RBX1 proteins to form an SCF(Fbxo7) E3 ubiquitin ligase complex. Fbxo7 negatively regulates NF-κB signaling through TRAF2 and cIAP1 ubiquitination. METHODS: We combine co-immunoprecipitation, ubiquitination in vitro and in vivo, cycloheximide chase assay, ubiquitin chain restriction analysis and microscopy to investigate interaction between Fbxo7 and overexpressed UXT-V2-HA. RESULTS: The Ubl domain of Fbxo7 contributes to interaction with UXTV2. This substrate is polyubiquitinated by SCF(Fbxo7) with K48 and K63 ubiquitin chain linkages in vitro and in vivo. This post-translational modification decreases UXT-V2 stability and promotes its proteasomal degradation. We further show that UXTV1, an alternatively spliced isoform of UXT, containing 12 additional amino acids at the N-terminus as compared to UXTV2, also interacts with and is ubiquitinated by Fbxo7. Moreover, FBXO7 knockdown promotes UXT-V2 accumulation, and the overexpression of Fbxo7-ΔF-box protects UXT-V2 from proteasomal degradation and enhances the responsiveness of NF-κB reporter. We find that UXT-V2 colocalizes with Fbxo7 in the cell nucleus. CONCLUSIONS: Together, our study reveals that SCF(Fbxo7) mediates the proteasomal degradation of UXT-V2 causing the inhibition of the NF-κB signaling pathway. GENERAL SIGNIFICANCE: Discovering new substrates of E3 ubiquitin-ligase SCF(Fbxo7) contributes to understand its function in different diseases such as cancer and Parkinson.Biotechnology and Biological Science Research Council [BB/J007846/1]

A child presenting with acute renal failure secondary to a high dose of indomethacin: a case report

<p>Abstract</p> <p>Introduction</p> <p>Acute renal failure caused by nonsteroidal anti-inflammatory drugs administered at therapeutic doses is generally mild, non-anuric and transitory. There are no publications on indomethacin toxicity secondary to high doses in children. The aim of this article is to describe acute renal failure secondary to a high dose of indomethacin in a child and to review an error in a supervised drug prescription and administration system.</p> <p>Case presentation</p> <p>Due to a medication error, a 20-day-old infant in the postoperative period of surgery for Fallot's tetralogy received a dose of 10 mg/kg of indomethacin, 50 to 100 times higher than the therapeutic dose. The child presented with acute, oligo-anuric renal failure requiring treatment with continuous venovenous renal replacement therapy, achieving complete recovery of renal function with no sequelae.</p> <p>Conclusion</p> <p>In order to reduce medication errors in critically ill children, it is necessary to develop a supervised drug prescription and administration system, with controls at various levels.</p

Evaluation of static and dynamic balance in athletes with anterior cruciate ligament injury – A controlled study

OBJECTIVES: Anterior cruciate ligament injury leads to adaptive responses to maintain postural control. However, there is no consensus regarding whether leg dominance also affects postural control in athletes with anterior cruciate ligament injury. The purpose of this study was to evaluate dynamic and static postural control among athletes with and without anterior cruciate ligament injury to the dominant leg. METHODS: Twenty-eight athletes, twenty-one males and seven females aged 15-45 years, were allocated to one of two groups: the anterior cruciate ligament injury group (26±3 years) or the control group without anterior cruciate ligament injury (25±6.5 years). All subjects performed one legged stance tests under eyes open and eyes closed conditions and squat and kick movement tests using a postural control protocol (AccuSwayPlus force platform, Massachusetts). The center of pressure displacement and speed were measured by the force platform. In addition, the distance traveled on the single-leg hop test was assessed as an objective measure of function. RESULTS: Significantly greater mediolateral sway was found under the eyes closed condition (p=0.04) and during squat movement (p=0.01) in the anterior cruciate ligament injury group than in the control group. Analysis of the single-leg hop test results showed no difference between the groups (p=0.73). CONCLUSION: Athletes with anterior cruciate ligament injury had greater mediolateral displacement of the center of pressure toward the dominant leg under the eyes closed condition and during squat movement compared to control athletes

Comprehensive Identification of Protein Substrates of the Dot/Icm Type IV Transporter of Legionella pneumophila

A large number of proteins transferred by the Legionella pneumophila Dot/Icm system have been identified by various strategies. With no exceptions, these strategies are based on one or more characteristics associated with the tested proteins. Given the high level of diversity exhibited by the identified proteins, it is possible that some substrates have been missed in these screenings. In this study, we took a systematic method to survey the L. pneumophila genome by testing hypothetical orfs larger than 300 base pairs for Dot/Icm-dependent translocation. 798 of the 832 analyzed orfs were successfully fused to the carboxyl end of β-lactamase. The transfer of the fusions into mammalian cells was determined using the β-lactamase reporter substrate CCF4-AM. These efforts led to the identification of 164 proteins positive in translocation. Among these, 70 proteins are novel substrates of the Dot/Icm system. These results brought the total number of experimentally confirmed Dot/Icm substrates to 275. Sequence analysis of the C-termini of these identified proteins revealed that Lpg2844, which contains few features known to be important for Dot/Icm-dependent protein transfer can be translocated at a high efficiency. Thus, our efforts have identified a large number of novel substrates of the Dot/Icm system and have revealed the diverse features recognizable by this protein transporter

c-Src Regulates Akt Signaling in Response to Ghrelin via β-Arrestin Signaling-Independent and -Dependent Mechanisms

The aim of the present study was to identify the signaling mechanisms to ghrelin-stimulated activation of the serine/threonine kinase Akt. In human embryonic kidney 293 (HEK293) cells transfected with GHS-R1a, ghrelin leads to the activation of Akt through the interplay of distinct signaling mechanisms: an early Gi/o protein-dependent pathway and a late pathway mediated by β-arrestins. The starting point is the Gi/o-protein dependent PI3K activation that leads to the membrane recruitment of Akt, which is phosphorylated at Y by c-Src with the subsequent phosphorylation at A-loop (T308) and HM (S473) by PDK1 and mTORC2, respectively. Once the receptor is activated, a second signaling pathway is mediated by β-arrestins 1 and 2, involving the recruitment of at least β-arrestins, c-Src and Akt. This β-arrestin-scaffolded complex leads to full activation of Akt through PDK1 and mTORC2, which are not associated to the complex. In agreement with these results, assays performed in 3T3-L1 preadipocyte cells indicate that β-arrestins and c-Src are implicated in the activation of Akt in response to ghrelin through the GHS-R1a. In summary this work reveals that c-Src is crucially involved in the ghrelin-mediated Akt activation. Furthermore, the results support the view that β-arrestins act as both scaffolding proteins and signal transducers on Akt activation

Estresse e Burnout entre residentes multiprofissionais

OBJECTIVE: To identify associations between high-stress and burnout syndrome in multidisciplinary residents from a federal university in Rio Grande do Sul, Brazil. METHOD: This is an analytical, cross-sectional and quantitative study. A socio-demographic questionnaire, the Work Stress Scale and the Maslach Burnout Inventory-Health Services Survey (MBI-HSS) were applied to 37 residents between April and June 2011. P-valuesOBJETIVO: Identificar la asociación entre alto estrés y Burnout en residentes Multiprofesionales de una universidad federal de Rio Grande do Sul. MÉTODO: se trata de un estudio analítico, transversal, cuantitativo. Se aplicaron un formulario de datos socio-demográficos, la Escala de Estrés en el Trabajo y el Maslach Burnout Inventory- Health Services en 37 residentes entre Abril y Junio de 2011. Valores de pOBJETIVO: identificar a associação entre alto estresse e Burnout em residentes multiprofissionais de uma universidade federal do Rio Grande do Sul. MÉTODO: trata-se de estudo analítico, transversal, quantitativo. Aplicaram-se um formulário de dados socio demográficos, a Escala de Estresse no Trabalho e o Maslach Burnout Inventory-Human Services Survey (MBI-HSS) em 37 residentes entre abril e junho de 2011. Valores de

Molecular Mimicry by an F-Box Effector of Legionella pneumophila Hijacks a Conserved Polyubiquitination Machinery within Macrophages and Protozoa

The ability of Legionella pneumophila to proliferate within various protozoa in the aquatic environment and in macrophages indicates a remarkable evolution and microbial exploitation of evolutionarily conserved eukaryotic processes. Ankyrin B (AnkB) of L. pneumophila is a non-canonical F-box-containing protein, and is the only known Dot/Icm-translocated effector of L. pneumophila essential for intra-vacuolar proliferation within both macrophages and protozoan hosts. We show that the F-box domain of AnkB and the 9L10P conserved residues are essential for intracellular bacterial proliferation and for rapid acquisition of polyubiquitinated proteins by the Legionella-containing vacuole (LCV) within macrophages, Dictyostelium discoideum, and Acanthamoeba. Interestingly, translocation of AnkB and recruitment of polyubiquitinated proteins in macrophages and Acanthamoeba is rapidly triggered by extracellular bacteria within 5 min of bacterial attachment. Ectopically expressed AnkB within mammalian cells is localized to the periphery of the cell where it co-localizes with host SKP1 and recruits polyubiquitinated proteins, which results in restoration of intracellular growth to the ankB mutant similar to the parental strain. While an ectopically expressed AnkB-9L10P/AA variant is localized to the cell periphery, it does not recruit polyubiquitinated proteins and fails to trans-rescue the ankB mutant intracellular growth defect. Direct in vivo interaction of AnkB but not the AnkB-9L10P/AA variant with the host SKP1 is demonstrated. Importantly, RNAi-mediated silencing of expression of SKP1 renders the cells non-permissive for intracellular proliferation of L. pneumophila. The role of AnkB in exploitation of the polyubiquitination machinery is essential for intrapulmonary bacterial proliferation in the mouse model of Legionnaires' disease. Therefore, AnkB exhibits a novel molecular and functional mimicry of eukaryotic F-box proteins that exploits conserved polyubiquitination machinery for intracellular proliferation within evolutionarily distant hosts

Screening non-coding RNAs in transcriptomes from neglected species using PORTRAIT: case study of the pathogenic fungus Paracoccidioides brasiliensis

<p>Abstract</p> <p>Background</p> <p>Transcriptome sequences provide a complement to structural genomic information and provide snapshots of an organism's transcriptional profile. Such sequences also represent an alternative method for characterizing neglected species that are not expected to undergo whole-genome sequencing. One difficulty for transcriptome sequencing of these organisms is the low quality of reads and incomplete coverage of transcripts, both of which compromise further bioinformatics analyses. Another complicating factor is the lack of known protein homologs, which frustrates searches against established protein databases. This lack of homologs may be caused by divergence from well-characterized and over-represented model organisms. Another explanation is that non-coding RNAs (ncRNAs) may be caught during sequencing. NcRNAs are RNA sequences that, unlike messenger RNAs, do not code for protein products and instead perform unique functions by folding into higher order structural conformations. There is ncRNA screening software available that is specific for transcriptome sequences, but their analyses are optimized for those transcriptomes that are well represented in protein databases, and also assume that input ESTs are full-length and high quality.</p> <p>Results</p> <p>We propose an algorithm called PORTRAIT, which is suitable for ncRNA analysis of transcriptomes from poorly characterized species. Sequences are translated by software that is resistant to sequencing errors, and the predicted putative proteins, along with their source transcripts, are evaluated for coding potential by a support vector machine (SVM). Either of two SVM models may be employed: if a putative protein is found, a protein-dependent SVM model is used; if it is not found, a protein-independent SVM model is used instead. Only <it>ab initio </it>features are extracted, so that no homology information is needed. We illustrate the use of PORTRAIT by predicting ncRNAs from the transcriptome of the pathogenic fungus <it>Paracoccidoides brasiliensis </it>and five other related fungi.</p> <p>Conclusion</p> <p>PORTRAIT can be integrated into pipelines, and provides a low computational cost solution for ncRNA detection in transcriptome sequencing projects.</p