Determinants of non-institutional childbirth: Evidence from the Peruvian demographic and health survey.

Objective: We aimed to evaluate the determinants of non-institutional delivery among women of childbearing age in Peru. Methods: We conducted a secondary analysis of data from the Peru 2019 Demographic and Family Health Survey (ENDES). This multi-stage survey sampling is representative at the urban-rural, regional, and national levels. The outcome variable was place of delivery, collected by self-reporting. Binary logistic regression models were used to assess the factors associated with non-institutionalized delivery. Thus, we estimated crude and adjusted odd ratios (aOR). For the multivariable model, the manual forward selection method and the Wald test were used to obtain a final parsimonious model. Results: The final sample included 14,061 women of reproductive age between the ages of 15 and 49. The prevalence of non-institutional delivery was 7.8 %. Multivariate regression analysis found that having a secondary education (aOR:0.48; 95 % confidence interval [CI]:0.39-0.58) or higher (aOR:0.57; 95 %CI:0.42-0.78); belonging to the second (aOR:0.26; 95 %CI:0.20-0.33), third (aOR:0.28; 95 %CI:0.21-0.38), fourth (aOR:0.21; 95 %CI:0.13-0.33), or fifth wealth quintile (aOR:0.15; 95 %CI:0.09-0.27); and suffering intimate partner violence (aOR:0.76; 95 %CI:0.64-0.91) were associated with lower odds of non-institutional delivery, while not having some type of health insurance (aOR:3.12; 95 %CI:2.47-3.95), living in a rural area (aOR:1.93; 95 %CI:1.54-2.42), and having had three or more deliveries (aOR:1.36; 95 %CI:1.07-1.72), were associated with higher odds of non-institutional delivery. Conclusions: We found that not having health insurance, residing in a rural area, and having had three or more deliveries were factors associated with non-institutional delivery in women of childbearing age. We propose that should focus public health strategies towards providing education to women about maternal health, and likewise, facilitating access to specialized health centers for rural populations.

Changing the guard-nuclear pore complex quality control.

The integrity of the nuclear envelope depends on the function of nuclear pore complexes (NPCs), transport channels that control macromolecular traffic between the nucleus and cytosol. The central importance of NPCs suggests the existence of quality control (QC) mechanisms that oversee their assembly and function. In this perspective, we emphasize the challenges associated with NPC assembly and the need for QC mechanisms that operate at various stages of an NPC's life. This includes cytosolic preassembly QC that helps enforce key nucleoporin-nucleoporin interactions and their ultimate stoichiometry in the NPC in addition to mechanisms that monitor aberrant fusion of the inner and outer nuclear membranes. Furthermore, we discuss whether and how these QC mechanisms may operate to sense faulty mature NPCs to facilitate their repair or removal. The so far uncovered mechanisms for NPC QC provide fertile ground for future research that not only benefits a better understanding of the vital role that NPCs play in cellular physiology but also how loss of NPC function and/or these QC mechanisms might be an input to aging and disease.

Hypertension and Histopathology Severity of Non-Alcoholic Fatty Liver Disease Among Adults with Obesity: A Cross-Sectional Study.

Background: Cardiovascular diseases are responsible for the majority of deaths resulting from non-alcoholic fatty liver disease (NAFLD). NAFLD is associated with hypertension and this is a key predictor of severe liver outcomes and an indicator of nonspecific portal fibrosis. Aim: To assess the association between hypertension and NAFLD severity. Methods: We conducted a secondary analysis of data from Peruvian adults with obesity and NAFLD who attended a Peruvian bariatric center. The severity of NAFLD was assessed using the Fatty Liver Inhibition of Progression algorithm / Steatosis, Activity and Fibrosis score. Hypertension was determined by either being recorded in the medical records or if the patient had a systolic pressure ≥ 140 mmHg or diastolic pressure ≥ 90 mmHg. To evaluate the association of interest, we calculated crude and adjusted prevalence ratios (aPR) using Poisson generalized linear models with logarithmic link function and robust variances. For the multivariable models, we adjusted for age, sex, physical activity and smoking. Results: Our study included 234 participants. The prevalence of hypertension was 19.2%, while the prevalence of severe NAFLD was 46.2%. After adjusting for confounders, the prevalence of hypertension was found to be significantly higher in the severe NAFLD group compared to the non-severe group (aPR = 1.33; 95% CI: 1.03-1.74). When stratified by the presence of metabolic syndrome (MetS), the association remained significant only in the group without MetS (aPR = 1.80; 95% CI: 1.05-3.11). Conclusion: We found an association between hypertension and severe NAFLD in adults with obesity, particularly in those without MetS.

Dynamic molecular mechanism of the nuclear pore complex permeability barrier.

Nuclear pore complexes (NPCs) mediate nucleocytoplasmic transport of specific macromolecules while impeding the exchange of unsolicited material. However, key aspects of this gating mechanism remain controversial. To address this issue, we determined the nanoscopic behavior of the permeability barrier directly within yeast S. cerevisiae NPCs at transport-relevant timescales. We show that the large intrinsically disordered domains of phenylalanine-glycine repeat nucleoporins (FG Nups) exhibit highly dynamic fluctuations to create transient voids in the permeability barrier that continuously shape-shift and reseal, resembling a radial polymer brush. Together with cargo-carrying transport factors the FG domains form a feature called the central plug, which is also highly dynamic. Remarkably, NPC mutants with longer FG domains show interweaving meshwork-like behavior that attenuates nucleocytoplasmic transport in vivo. Importantly, the bona fide nanoscale NPC behaviors and morphologies are not recapitulated by in vitro FG domain hydrogels. NPCs also exclude self-assembling FG domain condensates in vivo, thereby indicating that the permeability barrier is not generated by a self-assembling phase condensate, but rather is largely a polymer brush, organized by the NPC scaffold, whose dynamic gating selectivity is strongly enhanced by the presence of transport factors.

The chaperone DNAJB6 surveils FG-nucleoporins and is required for interphase nuclear pore complex biogenesis.

Biogenesis of nuclear pore complexes (NPCs) includes the formation of the permeability barrier composed of phenylalanine-glycine-rich nucleoporins (FG-Nups) that regulate the selective passage of biomolecules across the nuclear envelope. The FG-Nups are intrinsically disordered and prone to liquid-liquid phase separation and aggregation when isolated. How FG-Nups are protected from making inappropriate interactions during NPC biogenesis is not fully understood. Here we find that DNAJB6, a molecular chaperone of the heat shock protein network, forms foci in close proximity to NPCs. The number of these foci decreases upon removal of proteins involved in the early steps of interphase NPC biogenesis. Conversely, when this process is stalled in the last steps, the number of DNAJB6-containing foci increases and these foci are identified as herniations at the nuclear envelope. Immunoelectron tomography shows that DNAJB6 localizes inside the lumen of the herniations arising at NPC biogenesis intermediates. Loss of DNAJB6 results in the accumulation of cytosolic annulate lamellae, which are structures containing partly assembled NPCs, a feature associated with disturbances in NPC biogenesis. We find that DNAJB6 binds to FG-Nups and can prevent the aggregation of the FG region of several FG-Nups in cells and in vitro. Together, our data show that the molecular chaperone DNAJB6 provides quality control during NPC biogenesis and is involved in the surveillance of native intrinsically disordered FG-Nups.

Reversible protein aggregation as cytoprotective mechanism against heat stress.

Temperature fluctuation is one of the most frequent threats to which organisms are exposed in nature. The activation of gene expression programs that trigger the transcription of heat stress-protective genes is the main cellular response to resist high temperatures. In addition, reversible accumulation and compartmentalization of thermosensitive proteins in high-order molecular assemblies are emerging as critical mechanisms to ensure cellular protection upon heat stress. Here, we summarize representative examples of membrane-less intracellular bodies formed upon heat stress in yeasts and human cells and highlight how protein aggregation can be turned into a cytoprotective mechanism.

Acute Heat Stress Leads to Reversible Aggregation of Nuclear Proteins into Nucleolar Rings in Fission Yeast.

Upon acute heat stress (HS), overall mRNA transcription, processing, and export are inhibited, leading to cell growth arrest. However, how cells turn off mRNA metabolism is not fully understood. Here, we show that acute HS results in the segregation and aggregation of multiple nuclear and nucleolar proteins into ring-like structures located at the nucleolar periphery (nucleolar rings [NuRs]). NuRs sequester essential factors required for nuclear mRNA metabolism and nuclear pore complex function, as well as cell-cycle regulators. When cells are switched back to growing temperatures, NuRs disaggregate, and their components relocate to their functional environments in an Hsf1- and Hsp104-dependent manner, and concomitantly with the reinitiation of cell growth. These findings highlight the contribution of reversible protein aggregation to the inhibition of overall RNA-related activities in the nucleus and its functional relevance in the maintenance of cellular homeostasis during acute HS.

Impact of the COVID-19 pandemic on general surgery residency program in Peru: A cross-sectional study.

BACKGROUND: General surgey is a specialty of high demand and relevance. We aimed to collect the opinions of the residents and their tutors and heads of department, regarding the impact that this COVID-19 pandemic is having - and will probably have - on the training of future general surgeons in Peru. METHODS: We conducted a cross-sectional study in Lima, Peru. We surveyed residents of general surgery, as well as their tutors and heads of surgery departments from 14 Peruvian hospitals. RESULTS: The impact of COVID-19 was considered severe in approximately 60% of first-year residents, 100% of second-year residents, 40% of third-year residents and about 80% of attending physicians. The 68.8% of the residents considered that the loss of surgical training opportunities during the pandemic would negatively affect their job performance. In addition, as of 03/16/2020, no residents had performed more than 25 elective surgeries, trauma surgeries or laparoscopic procedures. All the participants (including tutors and heads of departments) highlighted the need to extend the residency period. CONCLUSION: The COVID-19 pandemic has affected the training of the general surgery residents. Deficiencies need to be identified in order to evaluate extending the period of the medical residency program in Peru.

Selective Nuclear Pore Complex Removal Drives Nuclear Envelope Division in Fission Yeast.

An important question in cell biology is how cellular organelles partition during cell division. In organisms undergoing closed mitosis, the elongation of an intranuclear spindle drives nuclear division, generating two identically sized nuclei [1, 2]. However, how the site of nuclear division is determined and the underlying mechanism driving nuclear envelope (NE) fission remain largely unknown. Here, using the fission yeast, we show that the microtubule bundler Ase1/PRC1 at the spindle midzone is required for the local concentration of nuclear pore complexes (NPCs) in the region of the NE in contact with the central spindle. As the spindle elongates during anaphase B, components of these NPCs are sequentially eliminated, and this is accompanied by the local remodeling of the NE. These two events lead to the eventual removal of NPCs and nuclear division. In the absence of importin α, NPCs remain stable in this region and no event of NE remodeling is observed. Consequently, cells fail to undergo nuclear division. Thus, our results highlight a new role of the central spindle as a spatial cue that determines the site of nuclear division and point to NPC removal as the triggering event.

Prevalencia del síndrome de fragilidad y factores asociados en adultos mayores / Prevalence of Frailty Syndrome and Associated Factors in Elderly Adults

Introducción: En Latinoamérica, la prevalencia del síndrome de fragilidad en adultos mayores no institucionalizados de 60 años a más oscila entre 7,7 y 39,3 por ciento. En Perú, pocos estudios han abordado este tema y sus factores asociados en poblaciones similares. Objetivo: Identificar la prevalencia de fragilidad y los factores asociados en adultos mayores pertenecientes a siete centros de atención primaria. Métodos: Estudio transversal analítico cuya variable desenlace fue la presencia de fragilidad, según criterios de Fried. Se incluyeron también datos sociodemográficos (para la ocupación se usó la clasificación SEE y Sem FyC), tamizaje de depresión (con la escala de Yesavage), grado de funcionalidad (con el índice de Barthel) y las comorbilidades (usando el índice Geriátrico de Comorbilidad y el índice de severidad de Greenfield). Resultados: Se evaluaron 116 adultos mayores, la prevalencia de fragilidad fue de 20,7 por ciento. Los factores asociados fueron tener dos o más enfermedades (RPa = 3,33; IC 95 por ciento: 1,04 - 10,68), tener depresión moderada (RPa = 6,50; IC 95 por ciento: 2,52 - 16,75) y dependencia moderada para la realización de AVD (RPa = 2,63; IC 95 por ciento: 2,63 - 14,15). Los factores protectores fueron haber tenido una ocupación clase III (RPa = 0,11; IC 95 por ciento: 0,02 - 0,54), IV (RPa = 0,07; IC 95 por ciento: 0,01 - 0,35) o V (RPa = 0,24; IC 95 por ciento: 0,09 - 0,60) y el vivir con algún familiar (RPa = 0,25; IC 95 por ciento: 0,08 - 0,74). Conclusión: Uno de cada cinco adultos mayores fue frágil. Los síntomas depresivos, tener dos o más comorbilidades y el ser dependiente para las actividades de la vida diaria fueron factores asociados a la fragilidad. Por su parte, trabajar en cargos no directivos y el vivir acompañado de algún familiar fueron factores protectores(AU)

Introduction: In Latin America, the prevalence of frailty syndrome in non-institutionalized elderly adults aged 60 years and older ranges from 7.7 to 39.3 percent. In Peru, few studies have addressed this topic and its associated factors in similar populations. Objective: To identify the prevalence of frailty and the associated factors in elderly adults from seven primary care centers. Methods: Analytical and cross-sectional study whose outcome variable was the presence of frailty, according to the Fried criteria. We also included sociodemographic data (for occupation, the classifications of the Spanish Society of Epidemiology [SEE in Spanish] and the Spanish Society of Family and Community Medicine [SEMFyC in Spanish] were used), depression screening (using the Yesavage scale), degree of functionality (using the Barthel index), and comorbidities (using the Geriatric Index of Comorbidity and Greenfield's severity index). Results: 116 older adults were evaluated. The prevalence of frailty was 20.7 percent. The associated factors were having two or more illnesses (RPa=3.33; 95 percent CI: 1.04-10.68), having moderate depression (RPa=6.50; 95 percent CI: 2.52-16.75), and moderate dependence for performing daily-life activities (RPa=2.63; 95 percent CI: 2.63-14.15). Protective factors were having had a class III occupation (RPa=0.11; 95 percent CI: 0.02-0.54), class IV (RPa=0.07; 95 percent CI: 0.01-0.35), or class V (RPa=0.24; 95 percent CI: 0.09-0.60), and living with a relative (RPa=0.25; 95 percent CI: 0.08-0.74). Conclusion: One in five elderly adults was frail. Depressive symptoms, having two or more comorbidities, and being dependent for daily-life activities were factors associated with frailty. On the other hand, working in non-managerial positions and living with a family member were protective factors(AU)

Depression among Peruvian adults with hypertension and diabetes: Analysis of a national survey.

BACKGROUND AND AIMS: Hypertension and diabetes are global health problems with an incidence that is rapidly increasing worldwide. Depression is the most frequent mental health co-morbidity and is considered an important risk factor for the development of both diseases. This study aimed to determine the prevalence and factors associated with depression among Peruvian adults with hypertension and diabetes. METHODS: We performed a secondary analysis of the 2017 Peru Demographic and Family Health Survey, using data from 10,566 adults aged 40 and older. The main outcome was depression assessed with the Patient Health Questionnaire-9 (PHQ-9). Other important variables were the diagnosis of hypertension or diabetes, sex, age, level of education, geographical region, wealth index, daily smoking, harmful alcohol consumption and physical disability. RESULTS: The overall prevalence of depression was 23.15% (CI 95%: 21.42%-24.88%) and among adults with hypertension and diabetes it was 34.96% (CI 95%: 29.33%-40.59%) and 35.56% (CI 95%: 24.49%-46.63%), respectively. Whereas the prevalence of depression among adults with type 2 diabetes mellitus (T2DM) was higher in the physical disability group (PRa: 1.28; CI 95%: 1.12-1.45), the prevalence of depression among adults with hypertension was higher in the female group (PRa: 1.36; CI 95%: 1.20-1.54) and among those with harmful alcohol consumption (PRa: 1.50; CI 95%: 1.01-2.24). CONCLUSION: Peru has a considerable burden of depression in the hypertensive and diabetic populations. While the variables positively associated with depression in hypertensive adults were female gender and harmful alcohol consumption, in adults with diabetes it was the presence of physical disability.

Nuclear Mechanics in the Fission Yeast.

In eukaryotic cells, the organization of the genome within the nucleus requires the nuclear envelope (NE) and its associated proteins. The nucleus is subjected to mechanical forces produced by the cytoskeleton. The physical properties of the NE and the linkage of chromatin in compacted conformation at sites of cytoskeleton contacts seem to be key for withstanding nuclear mechanical stress. Mechanical perturbations of the nucleus normally occur during nuclear positioning and migration. In addition, cell contraction or expansion occurring for instance during cell migration or upon changes in osmotic conditions also result innuclear mechanical stress. Recent studies in Schizosaccharomyces pombe (fission yeast) have revealed unexpected functions of cytoplasmic microtubules in nuclear architecture and chromosome behavior, and have pointed to NE-chromatin tethers as protective elements during nuclear mechanics. Here, we review and discuss how fission yeast cells can be used to understand principles underlying the dynamic interplay between genome organization and function and the effect of forces applied to the nucleus by the microtubule cytoskeleton.

A new role for the nuclear basket network.

Our view of the nuclear pore complexes (NPCs) as gateways between the nuclear and cytoplasmic compartments has been largely expanded in recent years. NPCs have now demonstrated roles in genome regulation and maintenance from single cells to multicellular organisms. Both NPC proteins as well as components of the NPC basket act as dynamic scaffolds for silencing factors, and chromatin and cell cycle regulators. Components of the NPC basket also couple mRNA production and export, and prevent the exit of unprocessed mRNAs from the nucleus. Our recent work describes a novel function of the fission yeast nuclear basket component - the translocated promoter region (TPR) nucleoporin Alm1 - in proper localization of the proteasome to the nuclear envelope. Here we discuss how regulation of proteasome localization to the nuclear envelope by Alm1 is key to maintain kinetochores homeostasis and proper chromosome segregation.

The fission yeast nucleoporin Alm1 is required for proteasomal degradation of kinetochore components.

Kinetochores (KTs) are large multiprotein complexes that constitute the interface between centromeric chromatin and the mitotic spindle during chromosome segregation. In spite of their essential role, little is known about how centromeres and KTs are assembled and how their precise stoichiometry is regulated. In this study, we show that the nuclear pore basket component Alm1 is required to maintain both the proteasome and its anchor, Cut8, at the nuclear envelope, which in turn regulates proteostasis of certain inner KT components. Consistently, alm1-deleted cells show increased levels of KT proteins, including CENP-CCnp3, spindle assembly checkpoint activation, and chromosome segregation defects. Our data demonstrate a novel function of the nucleoporin Alm1 in proteasome localization required for KT homeostasis.

Consumo de calcio en gestantes durante el segundo y tercer trimestre atendidas en el Hospital Nacional Almanzor Aguinaga Asenjo en el año 2013 / Consumption of calcium in pregnant during the second and third quarter served in the National Hospital Almanzor Aguinaga Asenjo in the year 2013

Introducción: El objetivo fue determinar la cantidad de consumo de calcio en gestantes durante el segundo y tercer trimestre atendidas en el hospital nacional Almanzor Aguinaga Asenjo en el año 2013. El estudio: Se incluyó a todas las gestantes atendidas en el hospital y que aceptaron voluntariamente. Se empleó un recordatorio de 24 horas para el registro de los alimentos consumidos. Hallazgos: Se evaluó a 100 gestantes. La edad promedio fue 31.59 años. La media del consumo de calcio fue de 1030.745 mg/día. Conclusiones: Existe un bajo consumo de calcio en dieta de las gestantes atendidas en el Hospital Nacional Almanzor Aguianga Asenjo, sin embargo, ahí un menor número de gestantes que tienen un buen consumo de calcio que toman tabletas de calcio y su dieta.

Introduction. The objective was to determine the amount of calcium intake in pregnant women during the second and third trimester treated at the National Hospital Almanzor Aguinaga Asenjo in 2013. The study: The study included all pregnant women in the hospital atentidas and volunteered. They use a reminder of 24 hours for the registration of food consumed. Findings: We evaluated 100 pregnant women. The average age was 31.59 years. The mean calcium in take was 1030.745 mg / day. Conclusions: Alow calcium intake in pregnant women treated at the National Hospital Almanzor Aguianga Asenjo.

Validación del índice pediátrico de mortalidad 2 (PIM2) en Argentina: un estudio prospectivo, multicéntrico, observacional / Validation of the Pediatric Index of Mortality 2 (PIM2) in Argentina: a prospective, multicenter, observational study

Introducción. El índice pediátrico de mortalidad 2 (Pediatric Index of Mortality 2; PIM2, por sus siglas en inglés) es uno de los puntajes más utilizados para la predicción de la mortalidad en pacientes ingresados en las Unidades de Cuidados Intensivos Pediátricos (UCIP) argentinas. El objetivo de este estudio fue validar el puntaje PIM2 en las UCIP integrantes del Programa de Calidad de Atención de la Sociedad Argentina de Terapia Intensiva. Población y métodos. Estudio multicéntrico, prospectivo, observacional, de corte transversal. Se incluyeron todos los pacientes de entre 1 mes y 16 años de edad, ingresados en las UCIP participantes entre el 01-01-2009 y el 31-122009. Se evaluó la discriminación y calibración del puntaje PIM2 en toda la población y en diferentes subgrupos (riesgo de mortalidad, edad, diagnósticos de ingreso). Resultados. Se incluyeron 2832 pacientes. El PIM2 predijo 246 muertes; sin embargo, fallecieron 297 pacientes (p <0,01). La razón de mortalidad estandarizada fue 1,20 (IC 95%: 1,01-1,43). El área bajo la curva ROC fue 0,84 (IC 95%: 0,82-0,86). Se detectaron diferencias estadísticamente significativas entre las muertes observadas y las predichas para toda la población y en los distintos intervalos de riesgo (χ² 71,02; df 8; p <0,001). También se detectaron diferencias estadísticamente significativas entre las muertes observadas y esperadas en los pacientes adolescentes (37/22, p= 0,03) y en aquellos ingresados con patología respiratoria (105/81, p= 0,03). Conclusiones. El puntaje PIM2 permite diferenciar adecuadamente los pacientes que sobreviven de aquellos que fallecen. Sin embargo, subvalora el riesgo de muerte en forma global, especialmente en los pacientes adolescentes y en aquellos ingresados por causa respiratoria. Es fundamental considerar estas diferencias al interpretar los resultados.

Introduction. The Pediatric Index of Mortality 2 (PIM2) is one of the most commonly used scoring systems to predict mortality in patients admitted to pediatric intensive care units (PICU) in Argentina. The objective of this study was to validate the PIM2 score in PICUs participating in the Quality of Care Program promoted by the Argentine Society of Intensive Care.Population and Methods. Multicenter, prospective, observational, cross-sectional study.All patients between 1 month and 16 years old admitted to participating PICUs between January 1st, 2009 and December 31st, 2009 were included. The discrimination and calibration of the PIM2 score were assessed in the entire population and in different subgroups (risk of mortality, age, diagnoses on admission).Results. Two thousand, eight hundred and thirty-two patients were included. PIM2 predicted 246 deaths; however, 297 patients died (p < 0.01). The standardized mortality ratio was 1.20 (95% confidence interval [CI]: 1.01-1.43). The area under the ROC curve was 0.84 (95% CI: 0.82-0.86). Statistically significant differences were detected between the observed and the predicted mortality for the entire population and for the different risk intervals (χ²: 71.02, df: 8, p < 0.001). Statistically significant differences were also found between observed and predicted mortality in adolescent patients (37/22, p = 0.03) and in those hospitalized due to respiratory disease (105/81, p = 0.03).Conclusions. The PIM2 score adequately discriminates survivors from non-survivors. However, it underscores the overall risk of death, especially in adolescent patients and those hospitalized due to respiratory disease. It is critical to take such differences into account when interpreting results.

Validation of the Pediatric Index of Mortality 2 (PIM2) in Argentina: a prospective, multicenter, observational study.

INTRODUCTION: The Pediatric Index of Mortality 2 (PIM2) is one of the most commonly used scoring systems to predict mortality in patients admitted to pediatric intensive care units (PICU) in Argentina. The objective of this study was to validate the PIM2 score in PICUs participating in the Quality of Care Program promoted by the Argentine Society of Intensive Care. POPULATION AND METHODS: Multicenter, prospective, observational, cross-sectional study. All patients between 1 month and 16 years old admitted to participating PICUs between January 1st, 2009 and December 31st, 2009 were included. The discrimination and calibration of the PIM2 score were assessed in the entire population and in different subgroups (risk of mortality, age, diagnoses on admission). RESULTS: Two thousand, eight hundred and thirty-two patients were included. PIM2 predicted 246 deaths; however, 297 patients died (p < 0.01). The standardized mortality ratio was 1.20 (95% confidence interval [CI]: 1.01-1.43). The area under the ROC curve was 0.84 (95% CI: 0.82-0.86). Statistically significant differences were detected between the observed and the predicted mortality for the entire population and for the different risk intervals (χ2: 71.02, df: 8, p < 0.001). Statistically significant differences were also found between observed and predicted mortality in adolescent patients (37/22, p = 0.03) and in those hospitalized due to respiratory disease (105/81, p = 0.03). CONCLUSIONS: The PIM2 score adequately discriminates survivors from non-survivors. However, it underscores the overall risk of death, especially in adolescent patients and those hospitalized due to respiratory disease. It is critical to take such differences into account when interpreting results.

A Lallzyme MMX-based rapid method for fission yeast protoplast preparation.

Fungal cells including yeasts are surrounded by cell wall that counteracts turgor pressure and prevents cell lysis. Many yeast experiments, including genetic manipulation of sterile strains, morphogenesis studies, nucleic acid isolation and many others, require mechanical breakage or enzymatic removal of the cell wall. Some of these experiments require the generation of live cells lacking cell walls, called protoplasts, that can be maintained in osmostabilized medium. Enzymatic digestion of cell wall proteoglycans is a commonly used method of protoplast preparation. Currently existing protocols for fission yeast cell wall digestion are time consuming and not very efficient. We developed a new rapid method for fission yeast protoplast preparation that relies on digesting cell walls with Lallzyme MMX commercial enzyme mix, which produces protoplasts from all cells in less than 10 min. We demonstrate that these protoplasts can be utilized in three commonly used fission yeast protocols. Thus, we provide the fission yeast community with a robust and efficient plasmid extraction method, a new protocol for diploid generation and an assay for protoplast recovery that should be useful for studies of morphogenesis. Our method is potentially applicable to other yeasts and fungi.