Generation of induced pluripotent stem cell lines IRMBi003-A and IRMBi003-B from a healthy donor to model Alzheimer's disease.

Induced Pluripotent Stem Cell (iPSC) lines derived from healthy individuals are helpful and essential tools for disease modelling. Here, we described the reprogramming of skin fibroblasts obtained from a healthy 59-year-old individual without Alzheimer's disease. The generated iPSC lines have a normal karyotype, expressed pluripotency markers, and demonstrated the ability to differentiate into the three germ layers. The iPSC lines will be used as controls to study Alzheimer's disease mechanisms.

Establishment of induced pluripotent stem cells IRMBi005-A from a patient with sporadic Alzheimer's disease.

Alzheimer's disease (AD) is a progressive neurological disorder and the most common form of dementia worldwide. Sporadic Alzheimer's disease (sAD) cases are the main forms, over 95% of AD cases, but still poorly understood. Thereby there is a crucial need to develop in vitro models for studying this multifactorial disorder. Here, we report the reprogramming of skin fibroblasts from a 57-years-old male donor. The new generated iPSC cell line has a normal karyotype and, is pluripotent since it demonstrates the ability to differentiate in vitro into the three germ layers. This iPSC line will be used to understand pathological mechanisms of sAD.

Physicians' Perception of Oral Nutritional Supplement Acceptance and Tolerability in Malnourished Outpatients: PerceptiONS Study.

Malnutrition is a common condition associated with various pathologies such as infections, neoplasms and digestive system disorders. Patients can be managed using different strategies, which include dietary modifications or oral nutritional supplements (ONS). It is important to promote good ONS adherence in order to attain clinical efficacy and cost-effectiveness. Several factors (amount, type, duration and tolerability) may have an impact on ONS adherence. PerceptiONS is a descriptive, cross-sectional observational study based on an ad hoc electronic survey designed to explore physicians' perception of malnourished outpatients prescribed ONS. The survey considered adherence, acceptance/satisfaction, tolerability and benefits within the context of Spain's healthcare system. The perceptions of 548 physicians regarding the experience of 2516 patients were analyzed. From the physicians' perspective, 57.11% of patients adhered to over 75% of the prescribed ONS. The organoleptic properties of ONS represented the aspect with the most positive impact on adherence, with smell (43.72%) ranking as the top characteristic. In general, patients were satisfied (90.10%) with the ONS, with their related benefits (88.51%) and their organoleptic properties (90.42%), and accepted ONS in their daily diet (88.63%). ONS improved patients' general condition (87.04%), quality of life (QoL) (81.96%) and vitality/energy (81.28%). Physicians would prescribe the same ONS again in 96.4% of the cases.

Generation of induced pluripotent stem cells (iPSCs) IRMBi002-A from an Alzheimer's disease patient carrying a D694N mutation in the APP gene.

Induced pluripotent stem cells (iPSC) were generated from skin fibroblasts obtained from a 58 year-old woman suffering from Alzheimer's disease and carrying a D694N mutation on Amyloid precursor protein (APP). Fibroblasts were reprogrammed into iPSC using the integration-free Sendai Virus which allows the expression of the Yamanaka factors. Verification of their pluripotency was achieved by demonstrating the expression of pluripotency markers and their differentiation potential into the three primary germ layers. The cells have the corresponding mutation and present a normal karyotype. The reported APP-D694N iPSC line may be used to model and study human AD pathology in vitro.

Generation of induced pluripotent stem cells (IRMBi001-A) from an Alzheimer's disease patient carrying a G217D mutation in the PSEN1 gene.

Induced pluripotent stem cells (iPSC) were generated from skin fibroblasts obtained from a 50 year-old patient suffering from Alzheimer's disease and carrying a G217D causal mutation on presenilin 1 (PSEN1). iPSCs were obtained following reprogramming using the integration-free Sendai Virus system which allows expression of the Yamanaka factors. Verification of their pluripotency was achieved by demonstrating the expression of pluripotency markers and their differentiation potential into the three primary germ layers. iPS cells carry the patient G217D mutation and present a normal karyotype. The reported PS1-G217D iPSC line may be used to model and study human AD pathology in vitro.

Aggregation Patterns, Sampling Plan, and Economic Injury Levels for the New Citrus Pest Delottococcus aberiae (Hemiptera: Pseudococcidae).

Delottococcus aberiae De Lotto (Hemiptera: Pseudococcidae) is the latest exotic mealybug species introduced in citrus in the Mediterranean basin. It causes severe distortion and size reduction on developing fruits. Due to its first interaction with citrus, D. aberiae economic thresholds are still unknown for this crop and the current Integrated Pest Management programs have been disrupted. The objectives of this study were to determine the aggregation patterns of D. aberiae in citrus, develop an efficient sampling plan to assess its population density, and calculate its Economic and Economic Environmental Injury Levels (EIL and EEIL, respectively). Twelve and 19 orchards were sampled in 2014 and 2015, respectively. At each orchard, population densities were monitored fortnightly in leaves, twigs, and fruit, and fruit damage was determined at harvest. Our results showed a clumped aggregation of D. aberiae in all organs with no significant differences between generations on fruit. Fruit damage at harvest was strongly correlated with fruit occupation in spring. Based on these results and using chlorpyrifos as the insecticide of reference, the EIL and EEIL were calculated as 7.1 and 12.1% of occupied fruit in spring, respectively. With all this, we recommend sampling 275 fruits using a binomial sampling method or alternatively, 140 fruits with an enumerative method bimonthly between petal fall and July.

Genetic susceptibility to Gilbert's syndrome in a valencian population; efficacy of the fasting test. / Susceptibilidad genética del síndrome de Gilbert en población valenciana; eficiencia del test de ayuno.

OBJECTIVE: To describe the populational distribution of the UGT1A1*28 variant (genetic variant code rs8175347) located in the promotor of the UGT gene and correlate its genotypes with the results of the fasting test, as well as its relationship with the biochemical disorder of Gilbert's syndrome (GS) in a Valencian population. PATIENTS AND METHODS: We studied the prevalence of the genotypes (TA)6/6 (TA)6/7 and (TA)7/7 of the deleterious variant rs8175347 in 144 patients with hyperbilirubinemia, 38 of whom had previously undergone the fasting test to diagnose GS, and in 150 control patients. By analysing the genomic region of the TATA box of the UGT1A1 gene promotor using Sanger sequencing, we established the correlation between the rs8175347 genotypes and the fasting test results and with the patients' biochemical disorders. RESULTS: The rate of heterozygosity of allele (TA)7 in the control population was 32% and increased to 87.59% among the patients with suspected GS. The rate of genotype TA7/7 was 81.94% among the patients with hyperbilirubinemia, compared with 11.33% in the control patients. The fasting test showed a 15.79% rate of false negatives and a 5.26% rate of false positives. CONCLUSIONS: The high frequency of allele (TA)7 among the Valencian control population, almost double the 5% reported for European control patients, confirms the high rate of GS reported in the Spanish population, without observing significant differences between the geographical ends of the country. The efficacy and reliability of the fasting test for the diagnosis of GS is questionable.

Sampling Methods for Detection and Monitoring of the Asian Citrus Psyllid (Hemiptera: Psyllidae).

The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama is a key pest of citrus due to its role as vector of citrus greening disease or "huanglongbing." ACP monitoring is considered an indispensable tool for management of vector and disease. In the present study, datasets collected between 2009 and 2013 from 245 citrus blocks were used to evaluate precision, sensitivity for detection, and efficiency of five sampling methods. The number of samples needed to reach a 0.25 standard error-mean ratio was estimated using Taylor's power law and used to compare precision among sampling methods. Comparison of detection sensitivity and time expenditure (cost) between stem-tap and other sampling methodologies conducted consecutively at the same location were also assessed. Stem-tap sampling was the most efficient sampling method when ACP densities were moderate to high and served as the basis for comparison with all other methods. Protocols that grouped trees near randomly selected locations across the block were more efficient than sampling trees at random across the block. Sweep net sampling was similar to stem-taps in number of captures per sampled unit, but less precise at any ACP density. Yellow sticky traps were 14 times more sensitive than stem-taps but much more time consuming and thus less efficient except at very low population densities. Visual sampling was efficient for detecting and monitoring ACP at low densities. Suction sampling was time consuming and taxing but the most sensitive of all methods for detection of sparse populations. This information can be used to optimize ACP monitoring efforts.

Insecticide sprays, natural enemy assemblages and predation on Asian citrus psyllid, Diaphorina citri (Hemiptera: Psyllidae).

The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama is considered a key citrus pest due to its role as vector of 'huanglongbing' (HLB) or citrus greening, probably the most economically damaging disease of citrus. Insecticidal control of the vector is still considered a cornerstone of HLB management to prevent infection and to reduce reinoculation of infected trees. The severity of HLB has driven implementation of intensive insecticide programs against ACP with unknown side effects on beneficial arthropod fauna in citrus agroecosystems. We evaluated effects of calendar sprays directed against this pest on natural enemy assemblages and used exclusion to estimate mortality they imposed on ACP populations in citrus groves. Predator exclusion techniques were used on nascent colonies of D. citri in replicated large untreated and sprayed plots of citrus during the four major flushing periods over 2 years. Population of spiders, arboreal ants and ladybeetles were independently assessed. Monthly sprays of recommended insecticides for control of ACP, adversely affected natural enemy populations resulting in reduced predation on ACP immature stages, especially during the critical late winter/early spring flush. Consequently, projected growth rates of the ACP population were greatest where natural enemies had been adversely affected by insecticides. Whereas, this result does not obviate the need for insecticidal control of ACP, it does indicate that even a selective regimen of sprays can impose as yet undetermined costs in terms of reduced biological control of this and probably other citrus pests.

[Participatory design guide for mental health promotion in prisons]. / Diseño participativo de una Guía para la Promoción de la Salud Mental en el medio penitenciario.

OBJECTIVE: [corrected] The main aim was to describe the issues and the participatory process required to design a Guide to promotemental health in prison through group activities. MATERIAL AND METHOD: We reviewed the bibliography, the mental health policies, the workshops about healthy mental habits, and a video about protection and risk factors. We identified the stakeholders and sought their points of view about the topics included in the Guide. We decided on the contents of the Guide and the incorporation of the health assets model and the perspectives provided by gender and cultural diversity. After the initial design of the modules and sessions, we started a pilot in the Prison of Valencia and the Prison of Zaragoza with women and men from different cultures, incorporating the suggested improvements, unifying contents and the discursive style. RESULTS: The guide is formed by: a preface, introduction, description, modules, sessions and evaluation. It has 6 modules and 19 sessions on: health and motivation; self-esteem; health and emotions; more assets to improve health: relax, positive thinking, keeping calm, communication and problem resolution; progress is possible: resiliency and starring in my own change. Each session consists of: activities (objectives, material, allocated time and development), theoretical material and tabbed sheets for activities. The guide is available in print and online versions. CONCLUSIONS: A guide has been elaborated with involved stakeholders and the opinion of the prison population.

Altered gene expression profile in cumulus cells of mature MII oocytes from patients with polycystic ovary syndrome.

STUDY QUESTION: Oocyte developmental competence is altered in patients with polycystic ovary syndrome (PCOS); is gene expression in cumulus cells (CCs) from mature metaphase II oocytes of patients with PCOS altered as well? SUMMARY ANSWER: Compared with CCs from non-PCOS patients, the gene expression profile of CCs isolated from mature oocytes of patients with PCOS present alterations that could explain the abnormal folliculogenesis and reduced oocyte competence in such patients. WHAT IS KNOWN ALREADY: Abnormal mRNA expression of several members of the insulin-like growth factor (IGF) family in CCs from PCOS patients was previously reported. Moreover, the whole transcriptome has been investigated in cultured CCs from PCOS patients. STUDY DESIGN, SIZE AND DURATION: This retrospective study included six PCOS patients diagnosed following the Rotterdam Criteria and six non-PCOS patients who all underwent ICSI for male infertility in the assisted reproduction technique (ART) Department of Montpellier University Hospital, between 2009 and 2011. PARTICIPANTS/MATERIALS, SETTING AND METHODS: CCs from PCOS and non-PCOS patients who underwent controlled ovarian stimulation (COS) were isolated mechanically before ICSI. Gene expression profiles were analysed using the microarray technology and the Significance Analysis of Microarray was applied to compare the expression profiles of CCs from PCOS and non-PCOS patients. MAIN RESULTS: The gene expression profile of CCs from patients with PCOS was significantly different from that of CCs from non-PCOS patients. Specifically, CCs from women with PCOS were characterized by abnormal expression of many growth factors, including members of the epidermal growth factor-like (EGFR, EREG and AREG) and IGF-like families (IGF1R, IGF2R, IGF2BP2 and IGFBP2), that are known to play a role in oocyte competence. In addition, mRNA transcripts of factors involved in steroid metabolism, such as CYP11A1, CYP1B1, CYP19A1 and CYP2B7P1, were deregulated in PCOS CCs, and this could explain the abnormal steroidogenesis observed in these women. Functional annotation of the differentially expressed genes suggests that defects in the transforming growth factor ß and estrogen receptors signalling cascades may contribute to the reduced oocyte developmental competence in patients with PCOS. LIMITATIONS AND REASONS FOR CAUTION: Owing to the strict selection criteria (similar age, weight and reasons for ART), this study included a small sample size (six cases and six controls), and thus, further investigations using a large cohort of patients are needed to confirm these results. WIDER IMPLICATIONS OF THE FINDINGS: This study opens a new perspective for understanding the pathogenesis of PCOS. STUDY FUNDING/COMPETING INTERESTS: This work was partially supported by a grant from the Ferring Pharmaceutical. The authors of the study have no competing interests to report. TRIAL REGISTRATION NUMBER: Not applicable.

Slow freezing and vitrification differentially modify the gene expression profile of human metaphase II oocytes.

BACKGROUND: Cryopreservation is now considered as an efficient way to store human oocytes to preserve fertility. However, little is known about the effects of this technology on oocyte gene expression. The aim of this study was to examine the effect of the two cryopreservation procedures, slow freezing and vitrification, on the gene expression profile of human metaphase II (MII) oocytes. METHODS: Unfertilized MII oocytes following ICSI failure were cryopreserved either by slow freezing or by the Cryotip method for vitrification. After thawing, total RNA was extracted and analyzed using Affymetrix Human Genome U133 Plus 2.0 GeneChip arrays. The gene expression profiles and associated biological pathways in slowly frozen/thawed and vitrified MII oocytes were determined and compared with those of non-cryopreserved MII oocytes used as controls. RESULTS: Both cryopreservation procedures negatively affected the gene expression profile of human MII oocytes in comparison with controls. However, slowly frozen and vitrified MI oocytes displayed specific gene expression signatures. Slow freezing was associated with down-regulation of genes involved in chromosomal structure maintenance (KIF2C and KIF3A) and cell cycle regulation (CHEK2 and CDKN1B) that may lead to a reduction in the oocyte developmental competence. In vitrified oocytes, many genes of the ubiquitination pathway were down-regulated, including members of the ubiquitin-specific peptidase family and subunits of the 26S proteasome. Such inhibition of the degradation machinery might stabilize the maternal protein content that is necessary for oocyte developmental competence. CONCLUSIONS: The low pregnancy rates commonly observed when using human MII oocytes after slow freezing-thawing may be explained by the alterations of the oocyte gene expression profile.

Transcriptome analysis reveals dialogues between human trophectoderm and endometrial cells during the implantation period.

BACKGROUND: Crosstalk between human trophectoderm (TE) and endometrial cells during the implantation window is a complex and not well-understood process. The aims of this study were (i) to evaluate the global gene expression profile in TE cells from Day 5 human blastocysts issued from IVF, (ii) to compare these data with the transcriptomic profile of endometrial cells in stimulated cycles for IVF and (iii) to identify potential early dialogues between maternal and embryonic cells during the implantation window. METHODS: Endometrial biopsies (n = 18) from normal responder patients were performed on the day of embryo transfer (Day 5 after human chorionic gonadotrophin administration). TE biopsies from five blastocysts donated for research purposes were mechanically extracted. DNA microarray analysis was carried out to identify the specific gene expression profiles and the biological pathways activated during the implantation window in endometrial and TE cells. RESULTS: Several cytokines (such as PDGFA, placenta growth factor, IGF2BP1 and IGF2BP3) were up-regulated in human TE cells, whereas some of the corresponding receptors (PDGFRA and KDR) were over-expressed in the receptive endometrium, suggesting that these molecules are involved in the early dialogue between blastocyst and maternal endometrial cells. In addition, several adhesion molecules and extracellular matrix proteins (MCAM, ITGAE and LAMA1) were also over-expressed in the TE, while others (ALCAM, CEACAM1, PECAM1, ITGB8 and LAMA2) were restricted to the receptive endometrium. CONCLUSION: The present study shows that several growth factors, cytokines, integrins and adhesion molecules are expressed in the TE and endometrium at the time of implantation. These results could contribute to the understanding of the mechanisms involved in the early dialogue between blastocyst and endometrium during implantation. Such results should be confirmed by further studies.

Tracking medfly predation by the wolf spider, Pardosa cribata Simon, in citrus orchards using PCR-based gut-content analysis.

The Mediterranean fruit fly, Ceratitis capitata (Wiedemann), which is often controlled chemically, is a major citrus pest in Spain; however, alternative biological control strategies such as those based on the conservation of polyphagous predators should be developed. The wolf spider, Pardosa cribata Simon, is an abundant predator found in citrus orchards in eastern Spain. In this study, we have evaluated polymerase chain reaction (PCR)-based techniques as a means of detecting C. capitata DNA remains in P. cribata specimens. To do so, two pairs of C. capitata species-specific primers were designed and tested. Primer specificity was tested on species closely related to C. capitata and with other pests and natural enemies present in citrus orchards. Medfly DNA was detectable in 100% of P. cribata from 0 to 12 h post ingestion for both primer pairs, decreasing to 37% at 96 h after prey ingestion for one pair of primers. DNA detectability half-lives were of 78.25 h and 78.08 h for each pair of primers but no statistical differences were found between them. Pardosa cribata specimens were field-collected daily after sterile C. capitata pupae had been deployed in the citrus orchard. Afterwards, the wolf spiders were analyzed and DNA remains of C. capitata were detected in 5% of them, with a peak of 15% coinciding with maximum C. capitata emergence. This study is the first to reveal the potential use of DNA markers to track medfly predation by P. cribata in citrus orchards and provides a new tool to estimate the potential role of this spider in biological-control conservation programs.

[Usefulness of a rapid serum interleukin-6 test combined with C-reactive protein to predict sepsis in newborns with suspicion of infection]. / Utilidad de un test rápido de interleuquina-6 sérico combinado con proteína C reactiva para predecir la sepsis en recién nacidos con sospecha de infección.

AIM: To evaluate the diagnostic value of interleukin-6 (IL-6) to predict the likelihood of neonatal sepsis in order to design an algorithm to decide antibiotic therapy. METHODS: IL-6 and C-reactive protein (CRP) were determined in 42 newborns with clinical suspicion of infection. Newborns were classified as a confirmed, probable or no infection, based on the results of cultures, chest X-rays and the involvement of four or more clinical areas on a scale of eight. Samples for IL-6 were collected in the initial assessment and frozen until its determination at the end of the study. Blinded IL-6 measurements were performed using a rapid test. Receiver operator characteristics curves (ROC) for CRP and IL-6 versus infection (confirmed or probable) were determined. RESULTS: Among the 42 cases included in the study 11 (26.2%) were classified as confirmed or probable infection. The area under curve (AUC) for IL-6 was 0.9, with a cut-off value of 53 pg/ml: sensitivity 90.91%, specificity 80%, positive predictive value (PPV) 62.5% and negative (NPV) 96% The level of IL-6>96 pg/ml and/or the combination of IL-6>53+CRP>13.3 mg/l, were the markers that best predicted infection: specificity 100% and PPV: 100%. CONCLUSIONS: Assessment of IL-6 could allow withholding or early discontinuation of antibiotics in newborns with IL-6<54 pg/ml. In cases with IL-6>96 pg/ml and/or IL-6>53+ CRP>13.3, antibiotics should be started promptly, given the high likelihood of infection. Implementation of an algorithm based on the determination of IL-6 and CRP, in the initial assessment of the newborn with clinical suspicion of infection, could reduce unnecessary antibiotic therapy.