Andrea Morales

Isolation and functional characterization of cold-regulated promoters, by digitally identifying peach fruit cold-induced genes from a large EST dataset

BackgroundCold acclimation is the process by which plants adapt to the low, non freezing temperatures that naturally occur during late autumn or early winter. This process enables the plants to resist the freezing temperatures of winter. Temperatures similar to those associated with cold acclimation are also used by the fruit industry to delay fruit ripening in peaches. However, peaches that are subjected to long periods of cold storage may develop chilling injury symptoms (woolliness and internal breakdown). In order to better understand the relationship between cold acclimation and chilling injury in peaches, we isolated and functionally characterized cold-regulated promoters from cold-inducible genes identified by digitally analyzing a large EST dataset.ResultsDigital expression analyses of EST datasets, revealed 164 cold-induced peach genes, several of which show similarities to genes associated with cold acclimation and cold stress responses. The promoters of three of these cold-inducible genes (Ppbec1, Ppxero2 and Pptha1) were fused to the GUS reporter gene and characterized for cold-inducibility using both transient transformation assays in peach fruits (in fruta) and stable transformation in Arabidopsis thaliana. These assays demonstrate that the promoter Pptha1 is not cold-inducible, whereas the Ppbec1 and Ppxero2 promoter constructs are cold-inducible.ConclusionThis work demonstrates that during cold storage, peach fruits differentially express genes that are associated with cold acclimation. Functional characterization of these promoters in transient transformation assays in fruta as well as stable transformation in Arabidopsis, demonstrate that the isolated Ppbec1 and Ppxero2 promoters are cold-inducible promoters, whereas the isolated Pptha1 promoter is not cold-inducible. Additionally, the cold-inducible activity of the Ppbec1 and Ppxero2 promoters suggest that there is a conserved heterologous cold-inducible regulation of these promoters in peach and Arabidopsis. These results reveal that digital expression analyses may be used in non-model species to identify candidate genes whose promoters are differentially expressed in response to exogenous stimuli.

Treatment of female stress urinary incontinence with Erbium-YAG laser in non-ablative mode

OBJECTIVE To evaluate the efficacy of laser photothermal therapy in a group of Chilean women with SUI. MATERIAL AND METHODS Longitudinal prospective study based on 42 women with mild-to-severe SUI, intervened with non ablative Er:YAG laser, between July 2014 and October 2015, in Santiago, Chile. The therapy efficacy was evaluated through the difference between every patients scores obtained, before and after treatment, with the International Consultation on Incontinence Questionnaire - Urinary Incontinence Short Form (ICIQ-SF), at a confidence level of 95%. Also, the patient satisfaction with treatment was reported through an ordinal scale. RESULTS ICIQ-SF median score was 11 before treatment and 3 after 6 months, with a significant difference per patient (p<0.001). 78.6% (n=33) reported improvement and 38.1% (n=16), a complete healing of SUI at follow up. 66.7% (n=28) reported high satisfaction and 81.8% (n=27) of sexually active women, also reported improvement of sexual gratification. Only mild pain during the procedure was reported as adverse effect. CONCLUSIONS Based on this short-term pilot study, non-ablative Er:YAG laser procedure seems to be a safe and efficacious alternative for patients with SUI. Further controlled studies will help to validate the use of non-ablative Er:YAG for treatment of SUI.

Multidimensional fluorescence microscopy of multiple organelles in Arabidopsis seedlings

BackgroundThe isolation of green fluorescent protein (GFP) and the development of spectral variants over the past decade have begun to reveal the dynamic nature of protein trafficking and organelle motility. In planta analyses of this dynamic process have typically been limited to only two organelles or proteins at a time in only a few cell types.ResultsWe generated a transgenic Arabidopsis plant that contains four spectrally different fluorescent proteins. Nuclei, plastids, mitochondria and plasma membranes were genetically tagged with cyan, red, yellow and green fluorescent proteins, respectively. In addition, methods to track nuclei, mitochondria and chloroplasts and quantify the interaction between these organelles at a submicron resolution were developed. These analyzes revealed that N-ethylmaleimide disrupts nuclear-mitochondrial but not nuclear-plastids interactions in root epidermal cells of live Arabidopsis seedlings.ConclusionWe developed a tool and associated methods for analyzing the complex dynamic of organelle-organelle interactions in real time in planta. Homozygous transgenic Arabidopsis (Kaleidocell) is available through Arabidopsis Biological Resource Center.

Transcriptional Responses of Chilean Quinoa (Chenopodium quinoa Willd.) Under Water Deficit Conditions Uncovers ABA-Independent Expression Patterns

HIGHLIGHTS R49 genotype displayed best performance on selected physiological parameters and highest tolerance to drought. R49 drought over-represented transcripts has exhibited 19% of genes (306 contigs) that presented no homology to published databases. Expression pattern for canonical responses to drought such as ABA biosynthesis and other genes induced in response to drought were assessed by qPCR. Global freshwater shortage is one of the biggest challenges of our time, often associated to misuse, increased consumption demands and the effects of climate change, paralleled with the desertification of vast areas. Chenopodium quinoa (Willd.) represents a very promising species, due to both nutritional content and cultivation under water constraint. We characterized drought tolerance of three Chilean genotypes and selected Genotype R49 (Salares ecotype) based upon Relative Water Content (RWC), Electrolyte Leakage (EL) and maximum efficiency of photosystem II (Fv/Fm) after drought treatment, when compared to another two genotypes. Exploratory RNA-Seq of R49 was generated by Illumina paired-ends method comparing drought and control irrigation conditions. We obtained 104.8 million reads, with 54 million reads for control condition and 51 million reads for drought condition. Reads were assembled in 150,952 contigs, were 31,523 contigs have a reading frame of at least 300 nucleotides (100 aminoacids). BLAST2GO annotation showed a 15% of genes without homology to NCBI proteins, but increased to 19% (306 contigs) when focused into drought-induced genes. Expression pattern for canonical drought responses such as ABA biosynthesis and other genes induced were assessed by qPCR, suggesting novelty of R49 drought responses.

Benefits of a root fungal endophyte on physiological processes and growth of the vulnerable legume tree Prosopis chilensis (Fabaceae)

Aims Desertification is a major concern in arid and semi-arid regions globally. Understanding interactions between vulnerable plant species and associated microbial symbionts may have important applications for conservation and restoration strategies in affected areas. Methods In this study, we evaluated the root-associated fungal endophyte community in Prosopis chilensis, a vulnerable and threatened aridadapted tree of northern Chile. Host benefits in terms of physiological performance and plant growth were also assessed. Endophytic fungi were isolated from asymptomatic roots by the culture method for molecular identification of the 18S rRNA gene. The dominant fungal endophyte (Penicillium sp.) in the community was later used in inoculation experiments to assess its effect on maximum quantum efficiency of photosystem II (PSII), (Fv/Fm) actual PSII efficiency (ΦPSII), and non-photochemical quenching (NPQ). Total sugars, starch content, malonaldehyde (MDA), nitrogen content (%) and growth traits were also measured. Important Findings Whereas ΦPSII increased significantly in endophyte-inoculated plants, NPQ was found to decrease. No effect of endophyte inoculation on sugars and MDA was detected, but starch content, leaf nitrogen content, number of leaves and shoot biomass were found to increase. Results revealed that inoculation of endophytic Penicillium fungal isolate can provide significant physiological benefits to the host plant P. chilensis. Its presence resulted in greater PSII efficiency and higher leaf nitrogen and carbohydrate content, enhancing host plant growth. These findings highlight the importance of considering the fungal endophyte community of this vulnerable species as an important tool to the design of further revegetation and conservation programs.

Perfil sociodemográfico y factores asociados a consultas por anticoncepción de emergencia

Background: Emergency contraception refers to contraceptive methods that can be used after a sexual intercourse Aim: To analyze the evolution of emergency contraception (EC) consultations on the Chilean public health system, at a communal level, in relation to the legal changes that recently took place, aimed to ease the delivery of the benefit. To analyze its association with socioeconomic, demographic and municipal healthcare system characteristics. Material and methods: We analyzed databases of the Ministry of Health to study EC consultation rates on Chilean communes, including consultations on emergency departments and by rape, from 2008 to 2013. We evaluated the association with communal characteristics, obtained from the National Municipal Information System. Results: Both the communal consultation rates and percentage of communes with consultations increased progressively during the period, with an explosive increase between 2009 and 2010. We observed a high dispersion in EC consultations both at a communal and regional level. There was an inverse significant association of the number of consultations with the communal poverty rate. Conclusions: Our results reveal the impact of legal modifications implemented in Chile since 2009 on the communal EC consultation rates. On other hand, our results reveal that although these modifications were oriented to favor the delivery of this benefit, a high dispersion subsists, associated with population’s socioeconomic factors, mainly, the communal poverty level.

Determinantes sociales de la edad de fallecimiento por causa cardiovascular

BACKGROUND The association of cardiovascular diseases with socio-demographic factors has not been fully explored. AIM To analyze the association of socio-demographic features with the survival time of individuals who died due to cardiovascular diseases. MATERIAL AND METHODS The death registries published by the Ministry of Health were analyzed. All deaths due to cardiovascular diseases occurred between 2001 and 2013 in people aged over one year were considered. Using a principal component analysis, the age at death was associated with socio-demographic features such as sex, marital status, residence zone, schooling, work status and medical care prior to death. RESULTS A total of 293,370 cardiovascular deaths were analyzed. The median age at death was 77 years. Not receiving medical care prior to death, was significantly associated with a lower age at the moment of the decease, mainly between 20 and 80 years of age. Among men, being occupationally active (hazard ratio (HR = 1.5 p < 0.01) and being unemployed (HR = 1.1 p < 0.01) were risks factors for early cardiovascular death. Being married, having higher schooling, residing in urban areas and having prior medical care were associated with a higher age at the moment of decease. CONCLUSIONS Socio-demographic profiles are associated with the age at death due to cardiovascular diseases. The effect of not receiving medical care on the age at death is noteworthy and reflects social inequities in the access to health care.

Perfil sociodemográfico y de riesgo cardiovascular en pacientes diabéticos de un consultorio urbano SSMN, 2004

Cardiovascular disease is of great relevance in Chile, resulting in high morbidity and mortality rates. Its control should include the overall management of the different cardiovascular risk factors. Amongst them, diabetes mellitus plays a crucial role. In order to describe a population of diabetic patients followed at one center of the Servicio de Salud Metropolitano Norte, we performed a retrospective descriptive study. Four hundred and thirty four clinical files from patients being followed up for diabetic foot between August 2003 and August 2004 were reviewed. Social and demographic variables (gender, age, marital status, level of education, occupation) as well as clinical variables (duration of diabetes, capillary glycemia, diagnosis of high arterial pressure, blood pressure, cholesterol levels, nutritional status, and alcohol and tobacco intake) were analyzed. The epidemiological parameters evidenced a predominantly female population, with a mean age of 61.3 years, most of them married, with basic scholar levels and unstable employments. Clinical variables showed an average duration of diabetic symptoms of 7 years, with abnormal glycemia, arterial blood pressure and cholesterol levels. Eighty eight (88%) percent had an altered nutritional status. Seventy eight percent (78%) of the sample had a high cardiovascular risk, regardless of diabetes mellitus. The observed clinical parameters raise concern, due to the deficient metabolic control of these patients and the presence of co-morbid conditions relevant to the etiology of cardiovascular disease. It is imperious to improve the control of cardiovascular risk factors among this group of patients, particularly emphasizing on educational support and ensuring the implementation of the prevailing regulations.