SUS1 introns are required for efficient mRNA nuclear export in yeast.

Efficient coupling between mRNA synthesis and export is essential for gene expression. Sus1/ENY2, a component of the SAGA and TREX-2 complexes, is involved in both transcription and mRNA export. While most yeast genes lack introns, we previously reported that yeast SUS1 bears two. Here we show that this feature is evolutionarily conserved and critical for Sus1 function. We determine that while SUS1 splicing is inefficient, it responds to cellular conditions, and intronic mutations either promoting or blocking splicing lead to defects in mRNA export and cell growth. Consistent with this, we find that an intron-less SUS1 only partially rescues sus1Δ phenotypes. Remarkably, splicing of each SUS1 intron is also affected by the presence of the other and by SUS1 exonic sequences. Moreover, by following SUS1 RNA and protein levels we establish that nonsense-mediated decay (NMD) pathway and the splicing factor Mud2 both play a role in SUS1 expression. Our data (and those of the accompanying work by Hossain et al.) provide evidence of the involvement of splicing, translation, and decay in the regulation of early events in mRNP biogenesis; and imply the additional requirement for a balance in splicing isoforms from a single gene.

A novel link between Sus1 and the cytoplasmic mRNA decay machinery suggests a broad role in mRNA metabolism.

BACKGROUND: Gene expression is achieved by the coordinated action of multiple factors to ensure a perfect synchrony from chromatin epigenetic regulation through to mRNA export. Sus1 is a conserved mRNA export/transcription factor and is a key player in coupling transcription initiation, elongation and mRNA export. In the nucleus, Sus1 is associated to the transcriptional co-activator SAGA and to the NPC associated complex termed TREX2/THSC. Through these associations, Sus1 mediates the nuclear dynamics of different gene loci and facilitate the export of the new transcripts. RESULTS: In this study, we have investigated whether the yeast Sus1 protein is linked to factors involved in mRNA degradation pathways. We provide evidence for genetic interactions between SUS1 and genes coding for components of P-bodies such as PAT1, LSM1, LSM6 and DHH1. We demonstrate that SUS1 deletion is synthetic lethal with 5'-->3' decay machinery components LSM1 and PAT1 and has a strong genetic interaction with LSM6 and DHH1. Interestingly, Sus1 overexpression led to an accumulation of Sus1 in cytoplasmic granules, which can co-localise with components of P-bodies and stress granules. In addition, we have identified novel physical interactions between Sus1 and factors associated to P-bodies/stress granules. Finally, absence of LSM1 and PAT1 slightly promotes the Sus1-TREX2 association. CONCLUSIONS: In this study, we found genetic and biochemical association between Sus1 and components responsible for cytoplasmic mRNA metabolism. Moreover, Sus1 accumulates in discrete cytoplasmic granules, which partially co-localise with P-bodies and stress granules under specific conditions. These interactions suggest a role for Sus1 in gene expression during cytoplasmic mRNA metabolism in addition to its nuclear function.

The mRNA export factor Sus1 is involved in Spt/Ada/Gcn5 acetyltransferase-mediated H2B deubiquitinylation through its interaction with Ubp8 and Sgf11.

Sus1 acts in nuclear mRNA export via its association with the nuclear pore-associated Sac3-Thp1-Cdc31 complex. In addition, Sus1 plays a role in transcription through its interaction with the Spt/Ada/Gcn5 acetyltransferase (SAGA) complex. Here, we have analyzed function and interaction of Sus1 within the SAGA complex. We demonstrate that Sus1 is involved in the SAGA-dependent histone H2B deubiquitinylation and maintenance of normal H3 methylation levels. By deletion analyses, we show that binding of Sus1 to SAGA depends on the deubiquitinylating enzyme Ubp8 and Sgf11. Moreover, a stable subcomplex between Sus1, Sgf11, and Ubp8 could be dissociated from SAGA under high salt conditions. In vivo recruitment of Sus1 to the activated GAL1 promoter depends on Ubp8 and vice versa. In addition, histones coenrich during SAGA purification in a Sus1-Sgf11-Ubp8-dependent way. Interestingly, sgf11 deletion enhances the mRNA export defect observed in sus1delta cells. Thus, the Sus1-Sgf11-Ubp8 module could work at the junction between SAGA-dependent transcription and nuclear mRNA export.

Postoperative Outcomes Within Enhanced Recovery After Surgery Protocol in Elective Total Hip and Knee Arthroplasty. POWER.2 Study: Study Protocol for a Prospective, Multicentre, Observational Cohort Study.

OBJECTIVE: The number of indications for total hip replacement (THR) and total knee replacement (TKR) surgery is increasing. Enhanced recovery after surgery (ERAS) represents the next step in the evolution of standardised care. The primary aim of this study is to measure the in-hospital 30-day medical and surgical postoperative complications rate. The study's secondary aims are to determine the length of stay, 30-day mortality rate, 30-day reoperation and readmission rates, the ERAS overall compliance and predefined ERAS individual items compliance. METHODS: This multicentre, prospective, observational study will include adult patients (aged >18 years) undergoing elective THR and TKR surgery. Consecutive patients undergoing surgery within the 2-month data collection period will be included. Centres that offer the THR and/or TKR surgery will be eligible to participate. The data collection will be done through an online data collection form via a secure, password-protected platform at each centre with predefined data fields. RESULTS: Ethical approval for this study has been obtained from the Comité de Ética de la Investigación de la Comunidad Autónoma de Aragón (C.P.-C.I. PI18/135; on 23 May 2018). It was prospectively registered on 27 June 2018, at www.clinicaltrials.gov with identification no. NCT03570944. CONCLUSION: The study will be disseminated through the SPARN-RedGERM, SEDAR, GERM and through social media. Peer-reviewed publications will be published under corporate authorship, including POWER.2 Study Group and SPARN-RedGERM.

[How to increase the health staff adherence to the hands hygiene protocol?] / ¿Cómo incrementar la adhesión del personal sanitario al protocolo de higiene de manos?

OBJECTIVE: Nosocomial infection is a frequent adverse event in health care. Hand hygiene (HH) reduces cross-transmission, but staff adherence is not always adequate. The objectives of this work were evaluating the effectiveness of the HH technique applied in the workplace and monitoring the adherence of the healthcare staff of a Hospital of Medium-Long Stay to the HH protocol. METHODS: Two cross sectional studies were carried out (2010-2012), to determine the prevalence of colonization of the hands of the staff by pathogenic microorganisms, a cross-sectional study (2015), to evaluate their knowledge of the HH technique and two cross-sectional studies (2015-2016), to evaluate the compliance of the protocol in the healthcare practice. The WHO Self-Assessment Framework was applied annually. The hydroalcoholic solution consumption was monitored as a process indicator. RESULTS: The prevalence of colonization of the staff hands went from 28.3% to 21.2%. Compliance with hand hygiene went from 39.5% to 72% and the results of the WHO Self-Assessment Framework went from an intermediate level with 287.5 points to an advanced level with 432.5 points. The consumption of hydroalcoholic solution went from 3.9 to 19.3 liters/1000 stays. CONCLUSIONS: Staff adherence to the HH protocol has increased, having significantly improved the compliance with the recommendations, the hydroalcoholic solution consumption and the HH level assigned to the hospital. The most effective measures to improve the results have been the development of specific improvement plans, having been decisive, the involvement of managers and the use of direct observation as a regular method of work.

OBJETIVO: La infección nosocomial es un evento adverso frecuente en la atención sanitaria. La higiene de manos (HM) reduce la transmisión cruzada, pero la adhesión del personal no siempre es adecuada. Los objetivos de este trabajo fueron evaluar la efectividad de la técnica de HM aplicada en el lugar de trabajo y monitorizar la adhesión del personal sanitario de un Hospital de Media-Larga Estancia al protocolo de HM. METODOS: Se realizaron dos cortes transversales (2010-2012), para conocer la prevalencia de colonización de las manos del personal por microorganismos patógenos, un estudio transversal (2015), para evaluar sus conocimientos sobre la técnica de HM y dos estudios observacionales (2015-2016) para evaluar el cumplimiento del protocolo en la práctica asistencial. Se aplicó anualmente el Marco de Autoevaluación de la OMS. Como indicador de proceso se monitorizó el consumo de solución hidroalcohólica. El análisis estadístico se realizó con el programa SPSS, versión 19.0. RESULTADOS: La prevalencia de colonización pasó del 28,3% al 21,2%. El cumplimiento de la HM pasó del 39,5% al 72%. Aplicando el Marco de Autoevaluación de la OMS se pasó de un nivel intermedio con 287,5 puntos a un nivel avanzado con 432,5 puntos. El consumo de solución hidroalcohólica pasó de 3,9 a 19,3 litros/1000 estancias. CONCLUSIONES: La adhesión al protocolo ha aumentado, habiendo mejorado significativamente el cumplimiento de las recomendaciones, el consumo de solución hidroalcohólica y el Nivel de HM asignado al hospital. Las medidas más eficaces para mejorar los resultados han sido la elaboración de planes de mejora específicos, habiendo sido decisiva la implicación de los directivos y el uso de la observación directa como método habitual de trabajo.

Rapid identification of RoHS-relevant flame retardants from polymer housings by ultrasonic extraction and RP-HPLC/UV.

A rapid method was developed for the extraction and identification of RoHS-relevant organic flame retardants in polymer material. Extraction was carried out using ultrasonic supported solvent extraction (USSE) and isopropanol. HPLC separation was achieved using a reversed-phase phenylhexyl-modified column and methanol, containing 2-aminoethanol-buffered, alkaline water. Identification was carried out by scanning UV detection and comparison with a library assembled from spectra of reference substances. The method was used to extract and identify polymer additives in TV and PC monitor housings. The overall runtime required for extraction and chromatographic analysis is less than 10 min. The limits of detection comply with the recommendations set by the German draft law.

¿Cómo incrementar la adhesión del personal sanitario al protocolo de higiene de manos? / How to increase the health staff adherence to the hands hygiene protocol?

Fundamentos: La infección nosocomial es un evento adverso frecuente en la atención sanitaria. La higiene de manos (HM) reduce la transmisión cruzada, pero la adhesión del personal no siempre es adecuada. Los objetivos de este trabajo fueron evaluar la efectividad de la técnica de HM aplicada en el lugar de trabajo y monitorizar la adhesión del personal sanitario de un Hospital de Media-Larga Estancia al protocolo de HM. Métodos: Se realizaron dos cortes transversales (2010-2012), para conocer la prevalencia de colonización de las manos del personal por microorganismos patógenos, un estudio transversal (2015), para evaluar sus conocimientos sobre la técnica de HM y dos estudios observacionales (2015-2016) para evaluar el cumplimiento del protocolo en la práctica asistencial. Se aplicó anualmente el Marco de Autoevaluación de la OMS. Como indicador de proceso se monitorizó el consumo de solución hidroalcohólica. El análisis estadístico se realizó con el programa SPSS, versión 19.0. Resultados: La prevalencia de colonización pasó del 28,3% al 21,2%. El cumplimiento de la HM pasó del 39,5% al 72%. Aplicando el Marco de Autoevaluación de la OMS se pasó de un nivel intermedio con 287,5 puntos a un nivel avanzado con 432,5 puntos. El consumo de solución hidroalcohólica pasó de 3,9 a 19,3 litros/1000 estancias. Conclusiones: La adhesión al protocolo ha aumentado, habiendo mejorado significativamente el cumplimiento de las recomendaciones, el consumo de solución hidroalcohólica y el Nivel de HM asignado al hospital. Las medidas más eficaces para mejorar los resultados han sido la elaboración de planes de mejora específicos, habiendo sido decisiva la implicación de los directivos y el uso de la observación directa como método habitual de trabajo

Background: Nosocomial infection is a frequent adverse event in health care. Hand hygiene (HH) reduces cross-transmission, but staff adhe-rence is not always adequate. The objectives of this work were evaluating the effectiveness of the HH technique applied in the workplace and monitoring the adherence of the healthcare staff of a Hospital of Medium-Long Stay to the HH protocol. Methods: Two cross sectional studies were carried out (2010-2012), to determine the prevalence of colonization of the hands of the staff by pathogenic microorganisms, a cross-sectional study (2015), to evaluate their knowledge of the HH technique and two cross-sectional studies (2015-2016), to evaluate the compliance of the protocol in the healthcare practice. The WHO Self-Assessment Framework was applied annually. The hydroalcoholic solution consumption was monitored as a process indicator. Results: The prevalence of colonization of the staff hands went from 28.3% to 21.2%. Compliance with hand hygiene went from 39.5% to 72% and the results of the WHO Self-Assessment Framework went from an intermediate level with 287.5 points to an advanced level with 432.5 points. The consumption of hydroalcoholic solution went from 3.9 to 19.3 liters/1000 stays. Conclusions: Staff adherence to the HH protocol has increased, ha-ving significantly improved the compliance with the recommendations, the hydroalcoholic solution consumption and the HH level assigned to the hospital. The most effective measures to improve the results have been the development of specific improvement plans, having been decisive, the involvement of managers and the use of direct observation as a regular method of work

Sus1 is recruited to coding regions and functions during transcription elongation in association with SAGA and TREX2.

Gene transcription, RNA biogenesis, and mRNA transport constitute a complicated process essential for all eukaryotic cells. The transcription/export factor Sus1 plays a key role in coupling transcription activation with mRNA export, and it resides in both the SAGA and TREX2 complexes. Moreover, Sus1 is responsible for GAL1 gene gating at the nuclear periphery, which is important for its transcriptional status. Here, we show that Sus1 is required during transcription elongation and is associated with the elongating form of RNA Polymerase II (RNAP II) phosphorylated on Ser5 and Ser2 of the C-terminal domain (CTD). In addition, Sus1 copurifies with the essential mRNA export factors Yra1 and Mex67, which bind to the mRNA cotranscriptionally. Consistently, ChIP analysis reveals that Sus1 is present at coding regions dependent on transcription in a manner stimulated by Kin28-dependent CTD phosphorylation. Strikingly, eliminating the TREX2 component Sac3 or the SAGA subunit Ubp8 partially impairs Sus1 targeting to coding sequences and upstream activating sequences (UAS). We found, unexpectedly, that Sgf73 is necessary for association of Sus1 with both SAGA and TREX2, and that its absence dramatically reduces Sus1 occupancy of UAS and ORF sequences. Our results reveal that Sus1 plays a key role in coordinating gene transcription and mRNA export by working at the interface between the SAGA and TREX2 complexes during transcription elongation.