Large 1p36 Deletions Affecting Arid1a Locus Facilitate Mycn-Driven Oncogenesis in Neuroblastoma.

Loss of heterozygosity (LOH) at 1p36 occurs in multiple cancers, including neuroblastoma (NBL). MYCN amplification and 1p36 deletions tightly correlate with markers of tumor aggressiveness in NBL. Although distal 1p36 losses associate with single-copy MYCN tumors, larger deletions correlate with MYCN amplification, indicating two tumor suppressor regions in 1p36, only one of which facilitates MYCN oncogenesis. To better define this region, we genome-edited the syntenic 1p36 locus in primary mouse neural crest cells (NCCs), a putative NBL cell of origin. In in vitro cell transformation assays, we show that Chd5 loss confers most of the MYCN-independent tumor suppressor effects of 1p36 LOH. In contrast, MYCN-driven tumorigenesis selects for NCCs with Arid1a deletions from a pool of NCCs with randomly sized 1p36 deletions, establishing Arid1a as the MYCN-associated tumor suppressor. Our findings reveal that Arid1a loss collaborates with oncogenic MYCN and better define the tumor suppressor functions of 1p36 LOH in NBL.

Cancer Mutations of the Tumor Suppressor SPOP Disrupt the Formation of Active, Phase-Separated Compartments.

Mutations in the tumor suppressor SPOP (speckle-type POZ protein) cause prostate, breast, and other solid tumors. SPOP is a substrate adaptor of the cullin3-RING ubiquitin ligase and localizes to nuclear speckles. Although cancer-associated mutations in SPOP interfere with substrate recruitment to the ligase, mechanisms underlying assembly of SPOP with its substrates in liquid nuclear bodies and effects of SPOP mutations on assembly are poorly understood. Here, we show that substrates trigger phase separation of SPOP in vitro and co-localization in membraneless organelles in cells. Enzymatic activity correlates with cellular co-localization and in vitro mesoscale assembly formation. Disease-associated SPOP mutations that lead to the accumulation of proto-oncogenic proteins interfere with phase separation and co-localization in membraneless organelles, suggesting that substrate-directed phase separation of this E3 ligase underlies the regulation of ubiquitin-dependent proteostasis.

Biodiversity loss along a gradient of deforestation in Amazonian agricultural landscapes.

Assessing how much management of agricultural landscapes, in addition to protected areas, can offset biodiversity erosion in the tropics is a central issue for conservation that still requires cross-taxonomic and landscape-scale studies. We measured the effects of Amazonia deforestation and subsequent land-use intensification in 6 agricultural areas (landscape scale), where we sampled plants and 4 animal groups (birds, earthworms, fruit flies, and moths). We assessed land-use intensification with a synthetic index based on landscape metrics (total area and relative percentages of land uses, edge density, mean patch density and diversity, and fractal structures at 5 dates from 1990 to 2007). Species richness decreased consistently as agricultural intensification increased despite slight differences in the responses of sampled groups. Globally, in moderately deforested landscapes species richness was relatively stable, and there was a clear threshold in biodiversity loss midway along the intensification gradient, mainly linked to a drop in forest cover and quality. Our results suggest anthropogenic landscapes with high-quality forest covering >40 % of the surface area may prevent biodiversity loss in Amazonia.

Fast-track Orthognathic Surgery: An Evidence-based Review.

The aim of this study was to establish a fast-track protocol for bimaxillary orthognathic surgery (OGS). Fast-track surgery (FTS) is a multidisciplinary approach where the pre-, intra-, and postoperative management is focusing maximally on a quick patient recovery and early discharge. To enable this, the patients' presurgical stress and postsurgical discomfort should be maximally reduced. Both referral patterns and expenses within the health-care system are positively influenced by FTS. University hospital-literature review through Medline, Embase, and the Cochrane Library (January 2000-July 2016) using the following words - "fast track, enhanced recovery, multimodal, and perioperative care" - to define a protocol evidence based for OGS, as well as evidenced-based medicine search of every term added to the protocol during the same period. The process has resulted in an OGS protocol that may improve the outcome of the patient through several nonoperative and operative measures such as preoperative patient education and intra/postoperative measures that should improve overall patient satisfaction, decrease morbidity such as postoperative nausea, headache, dizziness, pain, and intubation discomfort, and shorten hospital stay. A literature review allowed us to fine-tune a fast-track protocol for uncomplicated OGS that can be prospectively studied against currently applied ones.

Use of fused deposit modeling for additive manufacturing in hospital facilities: European certification directives.

PURPOSE: The goal of this study was to identify current European Union regulations governing hospital-based use of fused deposit modeling (FDM), as implemented via desktop three-dimensional (3D) printers. MATERIALS AND METHODS: Literature and Internet sources were screened, searching for official documents, regulations/legislation, and views of specialized attorneys or consultants regarding European regulations for 3D printing or additive manufacturing (AM) in a healthcare facility. A detailed review of the latest amendment (2016) of the European Parliament and Council legislation for medical devices and its classification was performed, which has regularly updated published guidelines for medical devices, that are classified by type and duration of patient contact. As expected, regulations increase in accordance with the level (I-III) of classification. RESULTS: Custom-made medical devices are subject to different regulations than those controlling serially mass-produced items, as originally specified in 98/79/EC European Parliament and Council legislation (1993) and again recently amended (2016). Healthcare facilities undertaking in-house custom production are not obliged to fully follow the directives as stipulated, given an exception for this scenario (Article 4.4a, 98/79/EC). CONCLUSION: Patient treatment and diagnosis with the aid of customized 3D printing in a healthcare facility can be performed without fully meeting the European Parliament and Council legislation if the materials used are ISO 10993 certified and article 4.4a applies.

Interaction networks and the use of floral resources by male orchid bees (Hymenoptera: Apidae: Euglossini) in a primary rain forests of the Chocó Region (Colombia).

Orchid bees are important keystone pollinators from the Neotropics. With the aim to study the relationships between orchid bees and their nectar and aromatic host species, we made systematic samplings of males across two conservation areas in the biogeographic Choc6 Region of Colombia. We used chemical baits to collect 352 male bees during five months. The pollen attached to their bodies was extracted for palynological identification and to estimate interaction networks. The euglossine community consisted of at least 22 species including Eg. maculilabris, Eg. orellana, Eg. championi and Eg. ignita. The male bees were associated with 84 plants but depended on a small group of them (Peperomia spp. and Anthurium spp, as well as species of Solanaceae, Ericaceae and Malpighiaceae) which were widely distributed across the altitudinal gradient, and were available through the year. The resulting interaction networks revealed a typical nested pattern usually found in plant-pollinator interactions, with several rare bee and plant species interaction with a small group of generalist bees and plant species. Albeit, we found variation within networks related to species composition. Such variation may be a consequence of specific differences in plant flowering phenology.

Interaction networks and the use of floral resources by male orchid bees (Hymenoptera: Apidae: Euglossini) in a primary rain forests of the Chocó Region (Colombia) / Redes de interacciones y uso de recursos florales por abejas macho de orquídeas (Hymenoptera: Apidae: Euglossini) en un bosque primario de la región del Chocó (Colombia)

Orchid bees are important keystone pollinators from the Neotropics. With the aim to study the relationships between orchid bees and their nectar and aromatic host species, we made systematic samplings of males across two conservation areas in the biogeographic Chocó Region of Colombia. We used chemical baits to collect 352 male bees during five months. The pollen attached to their bodies was extracted for palynological identification and to estimate interaction networks. The euglossine community consisted of at least 22 species including Eg. maculilabris, Eg. orellana, Eg. championiand Eg. ignita.The male bees were associated with 84 plants but depended on a small group of them (Peperomiaspp. and Anthuriumspp, as well as species of Solanaceae, Ericaceae and Malpighiaceae) which were widely distributed across the altitudinal gradient, and were available through the year. The resulting interaction networks revealed a typical nested pattern usually found in plant-pollinator interactions, with several rare bee and plant species interaction with a small group of generalist bees and plant species. Albeit, we found variation within networks related to species composition. Such variation may be a consequence of specific differences in plant flowering phenology.

Las abejas de las orquídeas son uno de los principales grupos de polinizadores con distribución exclusivamente Neotropical. Con el fin de documentar las relaciones de estas abejas con sus plantas fuente de néctar y sustancias aromáticas, realizamos muestreos sistemáticos de 352 machos durante cinco meses usando cebos químicos para atraerlos en dos áreas de conservación en el Chocó biogeográfico. Se extrajo el polen adherido al cuerpo de los especímenes recolectados para identificación palinológica de las especies vegetales visitadas por la comunidad y posterior análisis de redes de interacciones. Encontramos que la comunidad de euglossinos está conformada por al menos 22 especies de abejas. Dentro de la comunidad fueron más comunes: Eg. maculilabris, Eg. orellana, Eg. championiy Eg. ignita.Las especies de abejas se relacionan con no menos de 84 especies de plantas pero dependen más frecuentemente de un pequeño grupo de especies vegetales ampliamente distribuidas en el gradiente altitudinal mues-treado y que se encuentran disponibles durante gran parte del año. Dentro de este pequeño grupo destacan especies pertenecientes a los géneros Anthuriumy Peperomiay a las familias Solanaceae, Ericaceae y Malpighiaceae. Las redes de interacciones resultantes muestran un patrón anidado en el que muchas especies de abejas o plantas raras interac-túan con un pequeño grupo de especies de abejas o plantas generalistas. También encontramos variaciones espaciales y temporales en las redes en cuanto a la composición de especies y la manera como se distribuyen las interacciones. Estas variaciones estarían determinadas por las diferencias en la fenología de las plantas y en las condiciones climáticas entre los sitios muestreados que se encuentran muy cercanos entre sí.

Dissection of structural and functional requirements that underlie the interaction of ERdj3 protein with substrates in the endoplasmic reticulum.

ERdj3, a mammalian endoplasmic reticulum (ER) Hsp40/DnaJ family member, binds unfolded proteins, transfers them to BiP, and concomitantly stimulates BiP ATPase activity. However, the requirements for ERdj3 binding to and release from substrates in cells are not well understood. We found that ERdj3 homodimers that cannot stimulate the ATPase activity of BiP (QPD mutants) bound to unfolded ER proteins under steady state conditions in much greater amounts than wild-type ERdj3. This was due to reduced release from these substrates as opposed to enhanced binding, although in both cases dimerization was strictly required for substrate binding. Conversely, heterodimers consisting of one wild-type and one mutant ERdj3 subunit bound substrates at levels comparable with wild-type ERdj3 homodimers, demonstrating that release requires only one protomer to be functional in stimulating BiP ATPase activity. Co-expressing wild-type ERdj3 and a QPD mutant, which each exclusively formed homodimers, revealed that the release rate of wild-type ERdj3 varied according to the relative half-lives of substrates, suggesting that ERdj3 release is an important step in degradation of unfolded client proteins in the ER. Furthermore, pulse-chase experiments revealed that the binding of QPD mutant homodimers remained constant as opposed to increasing, suggesting that ERdj3 does not normally undergo reiterative binding cycles with substrates.

Modelo y cumplimentacion de alta hospitalaria de la unidad de gestión clínica de cirugía oral y maxilofacial. Cómo mejorarlo / Filling in the hospital discharge report in an oral and maxillofacial surgery clinical management unit. how to improve it

Objetivos. Conocer la calidad y grado de cumplimentación de la información recogida en las altas hospitalarias (IAH) del Servicio de Cirugía Oral y Maxilofacial de nuestro hospital. Material y métodos. Estudio descriptivo transversal de 152 altas hospitalarias. Incluye un total de 35 ítems agrupados. Para la evaluación del IAH valoramos la ausencia y el déficit de información. El criterio de adecuación se fijó por consenso en la cumplimentación del 90% de los ítems de forma correcta. Con el fin de conocer la fiabilidad de los resultados obtenidos se realizó la evaluación por dos revisores de forma independiente y en caso de discordancia se tomó la decisión por consenso tras revisar la historia clínica. Resultados. El 53,94% de los IAH disponen del 90% de los ítems completos y correctos del modelo de alta de nuestro Servicio. El 10,74% presentan alguna ausencia y el 65,79% de IAH presentan algún déficit de información. Se observa una variabilidad importante en la cumplimentación según el tipo de ítem analizado. Conclusiones. Nuestro trabajo valora la calidad de un modelo específico de IAH con ítems previamente seleccionados, considerados útiles y adecuados para reflejar de forma completa, exacta y precisa el proceso asistencial que recibe el paciente y detecta que los ítems: teléfono, residente, fechas de consulta, biopsia y de informe de biopsia, cirugía mayor ambulatoria e intervención urgente deben mejorarse(AU)

Objectives. To determine the quality and compliance to the information contained in the hospital discharge registry (HDR) issued by the Department of Oral and Maxillofacial Surgery in our hospital. Materials and Methods. A cross-sectional descriptive study was conducted to evaluate 152 HDR reports. These reports included a total of 35 items. We observed that there was a lack of information and some mistakes when filling the HDR boxes. Suitability criteria were set up by consensus as the completion of 90% of the items examined. To evaluate the reliability of the results, an assessment was performed by two reviewers independently, and in case of disagreement the decision was made by consensus after reviewing the medical record. Results. Only 53.94% of the HDR had 90% of the items completed appropriately; 10.74% had some items missing when filling in the form, and 65.79% lacked some information. Depending on the type of item there was a significant variation in the completion of the form. Conclusions. In the present work, we assess the quality of a specific hospital discharge form in our hospital, with items previously selected as useful and appropriate to reflect a complete, accurate and precise view of the care process of the patient, and improvements were needed in items such as, telephone, home address, date of consultation, biopsy report and the date it was performed, ambulatory surgery and emergency response(AU)

ERdj4 protein is a soluble endoplasmic reticulum (ER) DnaJ family protein that interacts with ER-associated degradation machinery.

Protein localization within cells regulates accessibility for interactions with co-factors and substrates. The endoplasmic reticulum (ER) BiP co-factor ERdj4 is up-regulated by ER stress and has been implicated in ER-associated degradation (ERAD) of multiple unfolded secretory proteins. Several other ERdj family members tend to interact selectively with nascent proteins, presumably because those ERdj proteins associate with the Sec61 translocon that facilitates entry of nascent proteins into the ER. How ERdj4 selects and targets terminally misfolded proteins for destruction remains poorly understood. In this study, we determined properties of ERdj4 that might aid in this function. ERdj4 was reported to retain its signal sequence and to be resistant to mild detergent extraction, suggesting that it was an integral membrane protein. However, live cell photobleaching analyses of GFP-tagged ERdj4 revealed that the protein exhibits diffusion coefficients uncommonly high for an ER integral membrane protein and more similar to the mobility of a soluble luminal protein. Biochemical characterization established that the ERdj4 signal sequence is cleaved to yield a soluble protein. Importantly, we found that both endogenous and overexpressed ERdj4 associate with the integral membrane protein, Derlin-1. Our findings now directly link ERdj4 to the ERAD machinery and suggest a model in which ERjd4 could help recruit clients from throughout the ER to ERAD sites.

Manejo del SAHS mediante dispositivos de avance mandibular. Estudio preliminar / Sleep apnea-hypopnea syndrome management using a mandibular advancement device. Preliminary study

El tratamiento que ha demostrado mayor eficacia en los pacientes con síndrome de apneas-hipopneas del sueño (SAHS) es la presión positiva continua de la vía aérea (CPAP). Los mayores inconvenientes son la incomodidad y la sensación de claustrofobia, que en algunos pacientes provoca rechazo o intolerancia. Una alternativa son los dispositivos de avance mandibular (DAM), que insertados en las arcadas dentarias producen el avance de la mandíbula y de la lengua, aumentando el volumen de la vía aérea. Objetivo: Presentar nuestra experiencia en el tratamiento del SAHS mediante dispositivos de avance mandibular tipo Herbst. Metodologia: Estudio de seguimiento prospectivo desde junio de 2006 hasta enero de 2009 de 7 pacientes del Área Hospitalaria Virgen Macarena con SAHS que rechazan el tratamiento con CPAP y a los que se ofrece tratamiento con DAM. Las variables analizadas son: índice de apneas-hipopneas por hora, índice de desaturaciones por hora, intensidad subjetiva del ronquido y el test de somnolencia de Epworth, antes del tratamiento y al menos 6 meses después desde el inicio de su uso. Utilizamos el test de Wilcoxon para detectar diferencias estadísticas significativas (p<0,05). Resultados: Se observó una reducción estadísticamente significativa del índice de apneahipopneas por hora (p<0,018) y del índice de desaturaciones por hora (p<0,018), así como una reducción no significativa del ronquido y de la somnolencia. Conclusiones: El uso de DAM tipo Herbst en pacientes afectos de SAHS que rechazan el uso de la CPAP podría ser útil, mejorando clínica y funcionalmente su situación(AU)

The most effective treatment in patients with sleep apnea-hypopnea syndrome (SAHS) is CPAP (continuous positive airway pressure). The main drawback of CPAP is the discomfort and claustrophobic sensation that it causes, which elicits rejection or intolerance by some patients. A non-surgical alternative to CPAP is the mandibular advancement device (MAD), which consists of a plastic splint inserted between the dental arches to shift the jaw and tongue forward and thus increase airway volume. Objective: Report our experience with the treatment of SAHS using the Herbst mandibular advancement device. Material and method: A prospective follow-up study was carried out from June 2006 until January 2009 at the Virgen Macarena University Hospital with 7 patients with SAHS who refused treatment with CPAP and were treated with the mandibular advancement device. The outcome variables analyzed were: apnea-hypopnea disruptions per hour index, desaturations per hour index, subjective intensity of snoring, and the Epworth Sleepiness Scale. Variables were evaluated pre-treatment and at least once 6 months after initiation MAD use. The Wilcoxon test for paired samples was used to detect statistically significant differences (p<0.05). Results: A statistically significant reduction in the hourly indices of apnea-hypopnea disruptions (p<0.018) and desaturations (p<0.018) was observed, as well as a statistically nonsignificant reduction in snoring and sleepiness. Conclusions: Use of the Herbst mandibular advancement device in patients with obstructive sleep apnea syndrome who refuse CPAP may be helpful as it improves the clinical and functional parameters of the condition(AU)

Proteasome nuclear import mediated by Arc3 can influence efficient DNA damage repair and mitosis in Schizosaccharomyces pombe.

Proteasomes must remove regulatory molecules and abnormal proteins throughout the cell, but how proteasomes can do so efficiently remains unclear. We have isolated a subunit of the Arp2/3 complex, Arc3, which binds proteasomes. When overexpressed, Arc3 rescues phenotypes associated with proteasome deficiencies; when its expression is repressed, proteasome deficiencies intensify. Arp2/3 is best known for regulating membrane dynamics and vesicular transport; thus, we performed photobleaching experiments and showed that proteasomes are readily imported into the nucleus but exit the nucleus slowly. Proteasome nuclear import is reduced when Arc3 is inactivated, leading to hypersensitivity to DNA damage and inefficient cyclin-B degradation, two events occurring in the nucleus. These data suggest that proteasomes display Arc3-dependent mobility in the cell, and mobile proteasomes can efficiently access substrates throughout the cell, allowing them to effectively regulate cell-compartment-specific activities.

Life and death of a BiP substrate.

BiP is the mammalian endoplasmic reticulum (ER) Hsp70 orthologue that plays a major role in all functions of this organelle including the seemingly opposing functions of aiding the maturation of unfolded nascent proteins and identifying and targeting chronically unfolded proteins for degradation. The recent identification of mammalian BiP co-factors combined with delineation of the ER degradation machinery and data suggesting that the ER is subdivided into unique regions helps explain how these different functions can occur in the same organelle and raises some unresolved issues.

Int6 and Moe1 interact with Cdc48 to regulate ERAD and proper chromosome segregation.

Int6/eIF3e is implicated in tumorigenesis, but its molecular functions remain unclear. We have studied its fission yeast homolog Yin6, reporting that it regulates proteolysis by controlling the assembly/localization of proteasomes, and binds directly to another conserved protein, Moe1. In the present study, we isolated Cdc48 as a Moe1-binding protein from a yeast two-hybrid screen, and confirmed biochemically that they form a stable complex in fission yeast. Overexpressing Moe1 or Yin6 partially rescued phenotypes of cdc48 mutants; conversely, overexpressing Cdc48 partially rescued phenotypes of moe1 or yin6 mutants. Mutants defective in both Cdc48 and the Yin6-Moe1 complex showed growth defects that were far more severe than either alone. These double mutants were severely deficient in endoplasmic reticulum associated degradation (ERAD), as they were hypersensitive to accumulation of misfolded proteins. In addition, their chromosomes showed frequent defects in spindle attachment and segregation--these mitotic defects correlated with Ase1 and Bir1/survivin mislocalization. These results suggest that Cdc48, Yin6 and Moe1 act in the same protein complex to concertedly control ERAD and chromosome segregation. Many of these properties are evolutionarily conserved in humans, since human Cdc48 rescued the lethality of the yeast cdc48Delta mutant, and Int6 and Moe1/eIF3d bind Cdc48 in human cells.

RPA-like proteins mediate yeast telomere function.

Cdc13, Stn1 and Ten1 are essential yeast proteins that both protect chromosome termini from unregulated resection and regulate telomere length. Cdc13, which localizes to telomeres through high-affinity binding to telomeric single-stranded DNA, has been extensively characterized, whereas the contribution(s) of the Cdc13-associated Stn1 and Ten1 proteins to telomere function have remained unclear. We show here that Stn1 and Ten1 are DNA-binding proteins with specificity for telomeric DNA substrates. Furthermore, Stn1 and Ten1 show similarities to Rpa2 and Rpa3, subunits of the heterotrimeric replication protein A (RPA) complex, which is the major single-stranded DNA-binding activity in eukaryotic cells. We propose that Cdc13, Stn1 and Ten1 function as a telomere-specific RPA-like complex. Identification of an RPA-like complex that is targeted to a specific region of the genome suggests that multiple RPA-like complexes have evolved, each making individual contributions to genomic stability.

Quaternary structure and cleavage specificity of a poxvirus holliday junction resolvase.

Recently, poxviruses were found to encode a protein with signature motifs present in the RuvC family of Holliday junction (HJ) resolvases, which have a key role in homologous recombination in bacteria. The vaccinia virus homolog A22 specifically cleaved synthetic HJ DNA in vitro and was required for the in vivo resolution of viral DNA concatemers into unit-length genomes with hairpin telomeres. It was of interest to further characterize a poxvirus resolvase in view of the low sequence similarity with RuvC, the absence of virus-encoded RuvA and RuvB to interact with, and the different functions of the viral and bacterial resolvases. Because purified A22 aggregated severely, studies were carried out with maltose-binding protein fused to A22 as well as to RuvC. Using gel filtration, chemical cross-linking, analytical ultracentrifugation, and light scattering, we demonstrated that A22 and RuvC are homodimers in solution. Furthermore, the dimeric form of the resolvase associated with HJ DNA, presumably facilitating the symmetrical cleavage of such structures. Like RuvC, A22 symmetrically cleaved fixed HJ junctions as well as junctions allowing strand mobility. Unlike RuvC and other members of the family, however, the poxvirus enzyme exhibited little cleavage sequence specificity. Structural and enzymatic similarities of poxvirus, bacterial, and fungal mitochondrial HJ resolvases are consistent with their predicted evolutionary relationship based on sequence analysis. The absence of a homologous resolvase in mammalian cells makes these microbial enzymes excellent potential therapeutic targets.

Efficient allelic exchange and transposon mutagenesis in Mycobacterium avium by specialized transduction.

Mycobacterium tuberculosis and Mycobacterium avium are pathogenic slow-growing mycobacteria that cause distinct human diseases. In contrast to recent advances in M. tuberculosis genetics and pathogenesis investigation, M. avium has remained genetically intractable and, consequently, its pathogenic strategies remain poorly understood. Here we report the successful development of efficient allelic exchange and transposon mutagenesis in an opaque clinical strain of M. avium by specialized transduction. Efforts to disrupt the leuD gene of M. avium by specialized transduction were successful but were complicated by inefficient isolation of recombinants secondary to high spontaneous antibiotic resistance. However, by using this leucine auxotroph as a genetic host and the Streptomyces coelicolor leuD gene as a selectable marker, we achieved efficient allelic exchange at the M. avium pcaA locus. A leuD-marked transposon delivered by specialized transduction mutagenized M. avium with efficiencies similar to M. tuberculosis. These results establish a system for random and directed mutagenesis of M. avium. In combination with the forthcoming M. avium genome sequence, these tools will allow the distinct physiologic and pathogenic properties of M. avium to be dissected in molecular detail.