Developmental trajectories of late preterm infants and predictors of academic performance.

BACKGROUND: The trajectories of late preterm development from infancy to kindergarten reading and math, and predictors of academic resilience and risk are unknown. METHODS: Sample included 1200 late preterm infants (LPIs) from the Early Childhood Longitudinal Study, Birth Cohort. Objective measurements of development at 9 and 24 months (Bayley-SFR) and reading and math academic achievement at preschool and kindergarten were standardized; trajectories of late preterm development from 9 months to kindergarten reading and math were identified using latent class growth analysis. Multinomial logistic regression [aOR, 95% CI] identified predictors of academic resilience and risk. RESULTS: Four trajectory groups were observed for reading and three for math. More optimal trajectories (in reading and math) and academic resilience were associated with experiencing sensitive parenting and preschool attendance. Suboptimal (at-risk) trajectories (in reading or math) and an increased odds of academic risk were associated with

In-the-moment ratings on the Early Relational Health Screen: A pilot study of application in home visiting and primary care.

Early infant-parent interaction sets a critical foundation for young children's well-being, and evidence regarding the protective role of secure early relationships has led to increased interest in effective screening and promotion of early relational health in pediatric primary care and home visiting settings. We report findings from two pilot studies conducted in the United States that describe the reliability and validity of a relational health screening tool, the Early Relational Health Screen (ERHS), implemented in two different contexts: an innovative model of relational health promotion in pediatric primary care (Study 1) and an Infant Mental Health Home Visiting (IMH-HV) model (Study 2). Across both studies, a trained clinician rated the ERHS following real-time observation of interaction (i.e., "in-the-moment" ratings). Reliability was assessed by comparing "in-the-moment" ERHS ratings to subsequent coding of the same interaction from video by an independent evaluator. In addition, Study 2 data permitted evaluation of the validity of "in-the-moment" ERHS ratings. Results from both studies indicated reliability of "in-the-moment" ERHS ratings. In addition, Study 2 clinician "in-the-moment" ratings were associated with maternal depression and ratings of child-parent interaction derived from a separate observational task coded by independent evaluators using a different well-validated research-based measure. Discussion highlights the potential of the ERHS as a screening, promotion, and prevention tool that may be feasibly administered by providers across pediatric primary care and home visiting settings.

La temprana interacción infante-progenitor establece una fundación esencial para el bienestar de los niños pequeños, y la evidencia sobre el papel de protección de tempranas relaciones receptivas ha aumentado el interés en la efectiva detección y promoción de la salud de la relación en el cuidado pediátrico primario y los escenarios de visitas a casa. Reportamos los resultados de dos estudios experimentales que describen la confiabilidad y validez de la Temprana Detección de la Salud de la Relación (ERHS) implementada en dos contextos: un modelo innovador de promoción de la salud de la relación en el cuidado primario (Estudio 1) y un modelo de salud mental infantil de visitas a casa (Estudio 2). A lo largo de ambos, un profesional clínico entrenado evaluó ERHS siguiendo una observación de interacción en tiempo real (v.g. puntajes asignados "en el momento"). Se evaluó la confiabilidad por medio de una comparación entre los puntajes del profesional clínico y los subsecuentes puntajes de la misma interacción en video por un evaluador independiente. Adicionalmente, los datos del Estudio 2 permitieron la evaluación de la validez de los puntajes de ERHS. Los resultados de ambos estudios indicaron la confiabilidad de los puntajes ERHS "en el momento." Es más, los puntajes del profesional clínico del Estudio 2 se asociaron con la depresión materna y los puntajes de la interacción niño-progenitor derivados de una tarea separada usando una medida bien validada basada en la investigación. Las discusiones subrayan el potencial de ERHS como una herramienta de detección, promoción y prevención que puede ser administrada factiblemente por los proveedores tanto en el cuidado primario como en los casos de visitas a casa.

L'interaction précoce nourrisson-parent jette les bases essentielles du bien-être du jeune enfant et l'évidence concernant le rôle protecteur des relations précoces sécures a mené à un intérêt plus grand pour le dépistage efficace et la promotion de la santé relationnelle précoce dans les soins de santé primaire en pédiatrie ainsi que les contextes de visites à domicile. Nous rapportons ici les résultats de deux études pilotes faites aux Etats-Unis d'Amérique, qui décrivent la fiabilité et la validité d'un outil de dépistage de la santé relationnelle, le Dépistage Précoce de Santé Relationnelle (en anglais Early Relational Health Screen dont nous gardons l'abréviation ici, ERHS), mis en place dans deux contextes différents: un modèle innovateur de promotion de la santé relationnelle précoce en soin pédiatrique primaire (Etude 1) et un modèle de visite à domicile pour la santé mentale du nourrisson (Etude 2). Au travers de ces deux études un clinicien entraîné a évalué l'ERHS après une observation en temps réel de l'interaction (c'est-à-dire, des scores "sur le moment"). La fiabilité a été évaluée en comparant l'ERHS "sur le moment" au codage ultérieur de la même interaction à partir d'une vidéo, par un évaluateur indépendant. De plus les données de l'Etude 2 ont permis l'évaluation de la validité des scores ERHS "sur le moment." Les résultats des deux études ont indiqué la fiabilité des scores ERHS "sur le moment." De plus les scores "sur le moment" du clinicien de l'Etude 2 étaient liés à la dépression maternelle et aux scores d'interaction enfant-parent dérivés d'une tâche observationnelle séparée codée par des évaluateurs indépendants en utilisant une mesure basée sur les recherches différente et communément validée. La discussion met en lumière le potentiel de l'ERHS en tant qu'outil de dépistage, de promotion et de prévention qui peut être réalistement utilisé par les professionnels au sein des soins primaires pédiatriques et des contextes de visites à domicile.

Multimodal assessments of preterm temperament across the first year of life: Continuity, stability, and moderation by country of origin and infant age.

There is limited research regarding the continuity, stability, and role of country of origin in preterm infant temperament across the first year of life. This prospective longitudinal study examined patterns of mean-level continuity and individual-differences stability of select scales of temperament at 6 and 12 months in preterm infants from three countries, Chile (n = 47), United Kingdom (n = 44), and United States (n = 50). Temperament was assessed with the Infant Behavior Questionnaire and observed using the Bayley Behavior Rating Scale. Continuity and stability across infant age, country effects, and interactions of country and age on preterm infant temperament were examined. Chilean mothers rated infants higher in soothability, duration of orienting, and orientation/engagement compared with mothers from the United Kingdom and/or United States. Continuity of temperament from 6 to 12 months varied by country: Chilean mothers reported increasing smiling and laughter and activity level from 6 to 12 months, and mothers from the United Kingdom reported decreasing smiling and laughter and increasing fear from 6 to 12 months. Infant temperament was stable in all three countries. Correlations evidenced low concordance between maternal reports and examiner observations of infant temperament at 12 months. However, among Chilean infants, higher maternal reported activity level was associated with higher examiner observed orientation/engagement score.

Hay una limitada investigación acerca de la continuidad, la estabilidad y el papel del país de origen en el temperamento de infantes nacidos prematuramente a lo largo del primer año de vida. Este potencial estudio longitudinal examinó patrones de continuidad en el promedio de nivel y las diferencias individuales en cuanto a la estabilidad de selectas escalas de temperamento a los 6 y 12 meses en infantes prematuros de tres países, Chile (n = 47), Reino Unido (n = 44) y Estados Unidos (n = 50). El temperamento se evaluó con el Cuestionario de Comportamiento del Infante y el mismo se observó usando la Escala de Puntuación del Comportamiento de Bayley. Se examinaron la continuidad y la estabilidad a lo largo de la edad del infante, los efectos del país, así como las interacciones entre país y edad en el temperamento de los infantes prematuros. Las madres chilenas evaluaron a sus infantes con más altos puntajes en cuanto a posibilidad de tranquilizarlos, duración de orientarlos y la orientación/participación en comparación con madres del Reino Unidos y/o de Estados Unidos. La continuidad de temperamento de los 6 a 12 meses varió según el país: las madres chilenas reportaron aumento en la sonrisa y la risa, y el nivel de actividad de los 6 a 12 meses, y las madres del Reino Unido reportaron una disminución en la sonrisa y la risa, y un aumento en el temor de los 6 a 12 meses. El temperamento del infante fue estable en los tres países. Las correlaciones son evidencia de la concordancia entre los reportes maternos y las observaciones del examinador del temperamento del infante a los 12 meses. Sin embrago, entre los infantes chilenos, el más alto nivel de actividad reportado por las madres se asoció con un más alto puntaje del examinador en cuanto a la observada orientación/participación.

Nous n'avons que des recherches limitées sur la continuité, la stabilité et le rôle du pays d'origine dans le tempérament du bébé prématuré au fil de la première année de la vie. Cette étude longitudinale prospective a examiné les modèles de continuité au niveau moyen et la stabilité des différences individuelles de certaines échelles de tempérament à 6 et à 12 mois chez les enfants prématurés de trois pays, le Chili (n = 47), le Royaume Uni (n = 44) et les Etats-Unis d'Amérique (n = 50). Le tempérament a été évalué au moyen du Questionnaire du Comportement du Nourrisson et observé en utilisant l'Echelle de Bailey d'Evaluation du Comportement du Nourrisson. La continuité et la stabilité au travers de l'âge du nourrisson, les effets du pays et les interactions du pays et de l'âge sur le tempérament du bébé prématuré ont été examinés. Les mères chiliennes ont évalué leurs bébés plus haut pour ce qui concernait la capacité à être calmé, la durée de l'orientation et l'orientation/l'engagement par comparaison aux mères du Royaume Uni et/ou des Etats-Unis. La continuité du tempérament de 6 à 12 mois a varié par pays: les mères chiliennes ont fait état de plus de sourires et de rires et d'un niveau d'activité plus élevé de 6 à 12 mois et les mères du Royaume Uni ont fait état d'une décroissance des sourires et des rires et d'une plus grande peur de 6 à 12 mois. Le tempérament du nourrisson était stable dans les trois pays. Les corrélations ont montré une concordance faible entre les rapports maternels et les observations de l'examinateur du tempérament du nourrisson à 12 mois. Cependant, chez les enfants chiliens, un niveau d'activité plus élevé rapporté par la mère était lié à un score d'orientation/d'engagement observé plus élevé de la part de l'examinateur.

Combining fluorescent cell barcoding and flow cytometry-based phospho-ERK1/2 detection at short time scales in adherent cells.

Detection of levels of intracellular phospho-proteins is key to analyzing the dynamics of signal transduction in cellular systems. Cell-to-cell variability in the form of differences in protein level in each cell affects signaling and is implicated in prognosis of many diseases. Quantitative analysis of such variability necessitate measuring the protein levels at single-cell resolution. Single-cell intracellular protein abundance detection in statistically significant number of adherent cells for short time sampling points post stimulation using classical flow cytometry (FCM) technique has thus far been a challenge due to the detrimental effects of cell detachment methods on the cellular machinery. We systematically show that cell suspension obtained by noninvasive temperature-sensitive detachment of adherent cells is amenable to high-throughput phospho-ERK1/2 protein detection at single-cell level using FCM in these short time sampling points. We demonstrate this on three adherent cell lines, viz., HeLa, A549, and MCF7, from distinct lineages having characteristically different elasticity at 37 °C. In particular, we use a right combination of multiplexing via fluorescent cell barcoding (FCB) and intracellular antibody staining for simultaneous detection of phospho-ERK1/2 (pERK) stimulated by epidermal growth factor (EGF) in multiple samples. Based on systematic characterization using Alexa 350 dye, we arrive at two conditions that must be satisfied for correct implementation of FCB. Our study reveals that the temperature-sensitive detachment of HeLa cells correctly captures the expected pronounced bimodal pERK distribution as an early response to EGF, which the enzymatic treatment methods fail to detect. © 2018 International Society for Advancement of Cytometry.

Bioassay for Determining the Concentrations of Caffeine and Individual Methylxanthines in Complex Samples.

Caffeine and other methylxanthines are stimulant molecules found in formulated beverages, including sodas and energy drinks, and in brewed beverages, such as coffee and teas. Previously, we developed a bioassay for caffeine that involves monitoring the growth of a ΔguaB mutant of Escherichia coli defective in de novo guanine biosynthesis. When supplemented with a plasmid expressing the genes for an N-demethylation pathway from Pseudomonas putida CBB5, these bacteria demethylate caffeine (1,3,7-trimethylxanthine) and other methylxanthines into xanthine, which is then converted into guanine to support cell growth. A major limitation of this bioassay was that it could only measure the total concentration of all methylxanthines in a mixture. Therefore, it could not be used to measure the caffeine content of beverages like teas, which contain substantial quantities of multiple methylxanthines. To overcome this limitation, we created seven new plasmids containing all subsets of the three demethylase genes (ndmA, ndmB, and ndmC). We show that strains of ΔguaBE. coli containing each plasmid are able to demethylate specific subsets of methylxanthines and that they can be used to determine the concentrations of individual methylxanthines in complex mixtures containing multiple methylxanthines, including coffee doped with an additional methylxanthine. While validating this assay, we also discovered an unexpected demethylation event at the 1-methyl position when NdmB and NdmC were expressed in the absence of NdmA. The improved cell-based bioassay is inexpensive, is easy to use, and gives results comparable to standard high-performance liquid chromatography methods for measuring methylxanthine concentrations.IMPORTANCE Caffeine (1,3,7-trimethylxanthine) is the dominant neurostimulant found in coffee, teas, sodas, and energy drinks. Measuring the amount of caffeine and other methylxanthines in these beverages is important for quality assurance and safety in food science. Methylxanthines are also used in medicine and as performance-enhancing drugs, two contexts in which accurately determining their concentrations in bodily fluids is important. Liquid chromatography is the standard method for measuring methylxanthine concentrations in a sample, but it requires specialized equipment and expertise. We improved a previous bioassay that links E. coli growth to methylxanthine demethylation so that it can now be used to determine the amounts of individual methylxanthines in complex mixtures or beverages, such as coffee.

Early childhood curiosity and kindergarten reading and math academic achievement.

BACKGROUND: Although children's curiosity is thought to be important for early learning, the association of curiosity with early academic achievement has not been tested. We hypothesized that greater curiosity would be associated with greater kindergarten academic achievement in reading and math. METHODS: Sample included 6200 children in the Early Childhood Longitudinal Study, Birth Cohort. Measures at kindergarten included direct assessments of reading and math, and a parent-report behavioral questionnaire from which we derived measures of curiosity and effortful control. Multivariate linear regression examined associations of curiosity with kindergarten reading and math academic achievement, adjusting for effortful control and confounders. We also tested for moderation by effortful control, sex, and socioeconomic status (SES). RESULTS: In adjusted models, greater curiosity was associated with greater kindergarten reading and math academic achievement: breading = 0.11, p < 0.001; bmath = 0.12, p < 0.001. This association was not moderated by effortful control or sex, but was moderated by SES (preading = 0.01; pmath = 0.005). The association of curiosity with academic achievement was greater for children with low SES (breading = 0.18, p < 0.001; bmath = 0.20, p < 0.001), versus high SES (breading = 0.08, p = 0.004; bmath = 0.07, p < 0.001). CONCLUSIONS: Curiosity may be an important, yet under-recognized contributor to academic achievement. Fostering curiosity may optimize academic achievement at kindergarten, especially for children with low SES.

Gestational Age and Kindergarten School Readiness in a National Sample of Preterm Infants.

OBJECTIVE: To examine the association of gestational age with school readiness in kindergarten reading and math skills. We hypothesized that compared with infants born at 39-41 weeks, infants born at lower gestational ages would have poorer school readiness. STUDY DESIGN: The study sample comprised 5250 children from the Early Childhood Longitudinal Study, Birth Cohort, assessed with specialized reading and math assessments at kindergarten. Poor school readiness was characterized by reading and math theta scores ≥1.5 SD below the sample mean. The aOR and 95% CI of poor school readiness were estimated using multivariate logistic regression, examining gestational age continuously and categorically (very preterm [VPT], moderate/late preterm [M/LPT], early term [ET], and term). Pairwise comparisons were performed to test for differences by gestational age category. RESULTS: There was an association between gestational age and poor school readiness for reading and math, with the suggestion of a threshold effect in children born at ≥32 weeks gestation. In adjusted models, in VPT infants, the aORs of poor school readiness in reading and math were 2.58 (95% CI, 1.29-5.15) and 3.38 (95% CI, 1.66-6.91), respectively. For infants born M/LPT and ET, the odds of poor school readiness in reading did not differ from those of children born full-term, however. CONCLUSIONS: Compared with term infants, the highest odds of poor school readiness in reading and math were seen in VPT infants, with lower odds of poor school readiness in children born at ≥32 weeks gestation. Ongoing developmental surveillance before kindergarten is indicated for VPT infants.

The Candida Genome Database: the new homology information page highlights protein similarity and phylogeny.

The Candida Genome Database (CGD, http://www.candidagenome.org/) is a freely available online resource that provides gene, protein and sequence information for multiple Candida species, along with web-based tools for accessing, analyzing and exploring these data. The goal of CGD is to facilitate and accelerate research into Candida pathogenesis and biology. The CGD Web site is organized around Locus pages, which display information collected about individual genes. Locus pages have multiple tabs for accessing different types of information; the default Summary tab provides an overview of the gene name, aliases, phenotype and Gene Ontology curation, whereas other tabs display more in-depth information, including protein product details for coding genes, notes on changes to the sequence or structure of the gene and a comprehensive reference list. Here, in this update to previous NAR Database articles featuring CGD, we describe a new tab that we have added to the Locus page, entitled the Homology Information tab, which displays phylogeny and gene similarity information for each locus.

The Aspergillus Genome Database: multispecies curation and incorporation of RNA-Seq data to improve structural gene annotations.

The Aspergillus Genome Database (AspGD; http://www.aspgd.org) is a freely available web-based resource that was designed for Aspergillus researchers and is also a valuable source of information for the entire fungal research community. In addition to being a repository and central point of access to genome, transcriptome and polymorphism data, AspGD hosts a comprehensive comparative genomics toolbox that facilitates the exploration of precomputed orthologs among the 20 currently available Aspergillus genomes. AspGD curators perform gene product annotation based on review of the literature for four key Aspergillus species: Aspergillus nidulans, Aspergillus oryzae, Aspergillus fumigatus and Aspergillus niger. We have iteratively improved the structural annotation of Aspergillus genomes through the analysis of publicly available transcription data, mostly expressed sequenced tags, as described in a previous NAR Database article (Arnaud et al. 2012). In this update, we report substantive structural annotation improvements for A. nidulans, A. oryzae and A. fumigatus genomes based on recently available RNA-Seq data. Over 26 000 loci were updated across these species; although those primarily comprise the addition and extension of untranslated regions (UTRs), the new analysis also enabled over 1000 modifications affecting the coding sequence of genes in each target genome.

Evolution of pathogenicity and sexual reproduction in eight Candida genomes.

Candida species are the most common cause of opportunistic fungal infection worldwide. Here we report the genome sequences of six Candida species and compare these and related pathogens and non-pathogens. There are significant expansions of cell wall, secreted and transporter gene families in pathogenic species, suggesting adaptations associated with virulence. Large genomic tracts are homozygous in three diploid species, possibly resulting from recent recombination events. Surprisingly, key components of the mating and meiosis pathways are missing from several species. These include major differences at the mating-type loci (MTL); Lodderomyces elongisporus lacks MTL, and components of the a1/2 cell identity determinant were lost in other species, raising questions about how mating and cell types are controlled. Analysis of the CUG leucine-to-serine genetic-code change reveals that 99% of ancestral CUG codons were erased and new ones arose elsewhere. Lastly, we revise the Candida albicans gene catalogue, identifying many new genes.

Improved gene ontology annotation for biofilm formation, filamentous growth, and phenotypic switching in Candida albicans.

The opportunistic fungal pathogen Candida albicans is a significant medical threat, especially for immunocompromised patients. Experimental research has focused on specific areas of C. albicans biology, with the goal of understanding the multiple factors that contribute to its pathogenic potential. Some of these factors include cell adhesion, invasive or filamentous growth, and the formation of drug-resistant biofilms. The Gene Ontology (GO) (www.geneontology.org) is a standardized vocabulary that the Candida Genome Database (CGD) (www.candidagenome.org) and other groups use to describe the functions of gene products. To improve the breadth and accuracy of pathogenicity-related gene product descriptions and to facilitate the description of as yet uncharacterized but potentially pathogenicity-related genes in Candida species, CGD undertook a three-part project: first, the addition of terms to the biological process branch of the GO to improve the description of fungus-related processes; second, manual recuration of gene product annotations in CGD to use the improved GO vocabulary; and third, computational ortholog-based transfer of GO annotations from experimentally characterized gene products, using these new terms, to uncharacterized orthologs in other Candida species. Through genome annotation and analysis, we identified candidate pathogenicity genes in seven non-C. albicans Candida species and in one additional C. albicans strain, WO-1. We also defined a set of C. albicans genes at the intersection of biofilm formation, filamentous growth, pathogenesis, and phenotypic switching of this opportunistic fungal pathogen, which provides a compelling list of candidates for further experimentation.

The Candida genome database incorporates multiple Candida species: multispecies search and analysis tools with curated gene and protein information for Candida albicans and Candida glabrata.

The Candida Genome Database (CGD, http://www.candidagenome.org/) is an internet-based resource that provides centralized access to genomic sequence data and manually curated functional information about genes and proteins of the fungal pathogen Candida albicans and other Candida species. As the scope of Candida research, and the number of sequenced strains and related species, has grown in recent years, the need for expanded genomic resources has also grown. To answer this need, CGD has expanded beyond storing data solely for C. albicans, now integrating data from multiple species. Herein we describe the incorporation of this multispecies information, which includes curated gene information and the reference sequence for C. glabrata, as well as orthology relationships that interconnect Locus Summary pages, allowing easy navigation between genes of C. albicans and C. glabrata. These orthology relationships are also used to predict GO annotations of their products. We have also added protein information pages that display domains, structural information and physicochemical properties; bibliographic pages highlighting important topic areas in Candida biology; and a laboratory strain lineage page that describes the lineage of commonly used laboratory strains. All of these data are freely available at http://www.candidagenome.org/. We welcome feedback from the research community at candida-curator@lists.stanford.edu.

The Aspergillus Genome Database (AspGD): recent developments in comprehensive multispecies curation, comparative genomics and community resources.

The Aspergillus Genome Database (AspGD; http://www.aspgd.org) is a freely available, web-based resource for researchers studying fungi of the genus Aspergillus, which includes organisms of clinical, agricultural and industrial importance. AspGD curators have now completed comprehensive review of the entire published literature about Aspergillus nidulans and Aspergillus fumigatus, and this annotation is provided with streamlined, ortholog-based navigation of the multispecies information. AspGD facilitates comparative genomics by providing a full-featured genomics viewer, as well as matched and standardized sets of genomic information for the sequenced aspergilli. AspGD also provides resources to foster interaction and dissemination of community information and resources. We welcome and encourage feedback at aspergillus-curator@lists.stanford.edu.

Periorbital Necrotizing Fasciitis: Presentation to Reconstruction.

Periorbital necrotizing fasciitis (NF) is a devastating bacterial infection associated with irreversible inflammatory destruction of soft tissues. Outcomes include disfigurement, vision loss, septic shock, and death within hours to days. We describe two cases of periorbital NF that presented to our unit within a three-month period. We aim to highlight the key clinical features of periorbital NF, demonstrate the rapid progression of the disease, and the need for prompt identification and decisive intervention. Both patients presented with fever and left-sided periorbital swelling and showed rapid progression of swelling and gangrenous changes to the periorbital skin with worsening proptosis. They were treated with broad-spectrum intravenous antibiotics and underwent emergency surgical debridement of necrotic tissue followed by reconstruction. We propose a formal protocol that we recommend to aid the diagnosis and management of periorbital NF in an acute setting.

Role of p53 transcription factor in determining the efficacy of telomerase inhibitors in cancer treatment.

AIM: Telomerase expression is unique to cancer cells, making it a promising target for therapy. However, a major drawback of telomerase inhibition is that it affects cancer cell proliferation only when telomeres shorten, creating a lag phase post-continuous drug treatment. Acute cytotoxicity of telomerase inhibitors is dependent on their ability to induce DNA damage. p53 senses DNA damage and is the primary effector required for sensitizing cells towards apoptosis. MAIN METHODS: Isogenic p53+/+ and p53-/- ovarian cancer cell lines were generated using the CRISPR/Cas9 system and the anti-cancer effect of telomerase inhibitors MST-312 and BIBR1532 were determined. Flow cytometry, real-time PCR, and western blot were performed to study cell cycle, apoptosis, and gene expression. KEY FINDINGS: We report that MST-312 exhibits p53-dependent cytotoxicity, while BIBR1532 exhibits p53-independent cytotoxicity. Colony-forming ability also confirms the p53-dependent effect of MST-312. Re-expression of p53 in p53-/- cells could rescue MST-312 sensitivity. In p53+/+ cells, MST-312 causes S phase arrest and activation of p53-dependent target genes like anti-apoptosis markers (Fas and Puma) and cell cycle markers (p21 and cyclinB). In p53-/- cells, MST-312 causes S/G2/M arrest. BIBR1532 induces S/G2/M phase cell cycle arrest irrespective of p53 status. This correlates with the expression of the DNA damage marker (γ-H2AX). Long-term continuous treatment with MST-312 or BIBR1532 results in p53-independent telomere shortening. SIGNIFICANCE: In summary, we demonstrate that acute anti-cancer effects of MST-312 are dependent on p53 expression. Hence, it is important to consider the p53 expression status in cancer cells when selecting and administering telomerase inhibitors.

The Outcomes of Involutional Ptosis Surgeries in a Single Tertiary Center.

PURPOSE: To report the surgical outcome of anterior approach primary ptosis surgery in a tertiary center and to compare redo surgical rates between different grades of surgeons. METHODS: This is a Retrospective review of series of annual audits. All involutional/aponeurosis-disinsertion ptosis surgeries performed at Moorfields Eye-hospital (MEH) between January 01, 2016 and December 31, 2019 were included. Only primary surgery was included. The following data were collected; number of surgeries per year, number of patients, demographics data, grades of surgeons, success rate, redo surgery rate from different grades of surgeons, complications rate and patients' satisfaction. RESULTS: During the study period, 1191 ptosis surgery were performed, with 899 (75%) cases being involutional/aponeurosis-disinsertion ptosis. The mean redo surgery rate within one year from the primary surgery was 10.5% and the mean complication rate was 1.0%, with 78.95% of patients reported being satisfied with the results of the surgery, having no difference between surgeon's grades. The redo surgery rate was higher for cases performed by a junior surgeon (fellow/registrar) (64.26%) than by a consultant (38.94%). CONCLUSIONS: We report the success rate of a large cohort of primary involutional ptosis surgery performed at the ophthalmic-specialist tertiary center. The success and complication rates are comparable to the literature at 90% and 1%, respectively. Redo surgeries were more frequently required when performed by junior surgeons compared to the consultants, whereas the patient satisfaction level did not differ between different grades of surgeons.

Comprehensive annotation of secondary metabolite biosynthetic genes and gene clusters of Aspergillus nidulans, A. fumigatus, A. niger and A. oryzae.

BACKGROUND: Secondary metabolite production, a hallmark of filamentous fungi, is an expanding area of research for the Aspergilli. These compounds are potent chemicals, ranging from deadly toxins to therapeutic antibiotics to potential anti-cancer drugs. The genome sequences for multiple Aspergilli have been determined, and provide a wealth of predictive information about secondary metabolite production. Sequence analysis and gene overexpression strategies have enabled the discovery of novel secondary metabolites and the genes involved in their biosynthesis. The Aspergillus Genome Database (AspGD) provides a central repository for gene annotation and protein information for Aspergillus species. These annotations include Gene Ontology (GO) terms, phenotype data, gene names and descriptions and they are crucial for interpreting both small- and large-scale data and for aiding in the design of new experiments that further Aspergillus research. RESULTS: We have manually curated Biological Process GO annotations for all genes in AspGD with recorded functions in secondary metabolite production, adding new GO terms that specifically describe each secondary metabolite. We then leveraged these new annotations to predict roles in secondary metabolism for genes lacking experimental characterization. As a starting point for manually annotating Aspergillus secondary metabolite gene clusters, we used antiSMASH (antibiotics and Secondary Metabolite Analysis SHell) and SMURF (Secondary Metabolite Unknown Regions Finder) algorithms to identify potential clusters in A. nidulans, A. fumigatus, A. niger and A. oryzae, which we subsequently refined through manual curation. CONCLUSIONS: This set of 266 manually curated secondary metabolite gene clusters will facilitate the investigation of novel Aspergillus secondary metabolites.