FInCH: FIJI plugin for automated and scalable whole-image analysis of protein expression and cell morphology.

Study of morphogenesis and its regulation requires analytical tools that enable simultaneous assessment of processes operating at cellular level, such as synthesis of transcription factors (TF), with their effects at the tissue scale. Most current studies conduct histological, cellular and immunochemical (IHC) analyses in separate steps, introducing inevitable biases in finding and alignment of areas of interest at vastly distinct scales of organization, as well as image distortion associated with image repositioning or file modifications. These problems are particularly severe for longitudinal analyses of growing structures that change size and shape. Here we introduce a python-based application for automated and complete whole-slide measurement of expression of multiple TFs and associated cellular morphology. The plugin collects data at customizable scale from the cell-level to the entire structure, records each data point with positional information, accounts for ontogenetic transformation of structures and variation in slide positioning with scalable grid, and includes a customizable file manager that outputs collected data in association with full details of image classification (e.g., ontogenetic stage, population, IHC assay). We demonstrate the utility and accuracy of this application by automated measurement of morphology and associated expression of eight TFs for more than six million cells recorded with full positional information in beak tissues across 12 developmental stages and 25 study populations of a wild passerine bird. Our script is freely available as an open-source Fiji plugin and can be applied to IHC slides from any imaging platforms and transcriptional factors.

Calidad de la dieta "antes y durante" un tratamiento de pérdida de peso basado en dieta mediterránea, terapia conductual y educación nutricional / Quality of the diet "before and during" a weight loss treatment based on mediterranean diet, behaviorual therapy and nutritional education

Introducción: El método de pérdida de peso (dieta mediterránea, terapia de comportamiento y educación nutricional) ha mostrado ser efectivo en el tratamiento de la obesidad. Objetivo: El objetivo del presente trabajo es evaluar y comparar la calidad de las dietas ingeridas antes y durante el tratamiento mediante el Índice de Alimentación Saludable (IAS) y su relación con diferentes variables. Materiales y métodos: La muestra fue de 392 pacientes (330 mujeres, 62 hombres), edad 39,3 ± 11,5 años y IMC de 31,2 ± 5,3 kg/m2. A partir del recuerdo-24 h previo al tratamiento y del registro dietético 7 días se estimó el IAS de “antes” y “durante” tratamiento. El IAS consta de 10 variables que representan el cumplimiento de objetivos nutricionales para la población española (SENC, 2004). Resultados: Dieta previa, presentó un IAS “necesita mejorar” (68,6 ± 11,6) con lípidos (%) (43,9 ± 8,4) y AGS (% lípidos) (67,4 ± 20,1) elevados, además el contenido en AGM (% lípidos) (27,8 ± 15,1) fue insuficiente. El IAS varió en función del IMC siendo el de obesos inferior al de personas con sobrepeso (65,1 ± 11,6 vs 69, 2 ± 13,9; P < 0,05). La dieta ingerida durante el tratamiento mejoró notablemente IAS (91,4 ± 9,7). El IAS de las mujeres fue superior (92,3 ± 9,1) al de los hombres (84,4 ± 12,0) (P < 0,05). Aquellos que alcanzaron la meta de pérdida de peso adquirieron mejores valores de IAS durante el tratamiento que los que no la alcanzaron (92,1 ± 9,2 vs 87,9 ± 11,7) (P < 0,05). Conclusiones: Según el IAS, la calidad de la dieta estudiada durante el tratamiento de pérdida de peso mejoró significativamente en relación a la dieta habitual del paciente. El IAS de la dieta durante el tratamiento se asocia con el sexo, el estado ponderal (sobrepeso y obesidad) y con el éxito del tratamiento (> 5% de pérdida del peso inicial) (AU)

INTRODUCTION: The "Garaulet" Method (Mediterranean Diet, therapy of behavior and nutritional education), has shown to be effective in the treatment of the obesity. OBJECTIVES: To evaluate and to compare the quality of the diets ingested before and during the treatment by means of Index-of-Feeding-Healthful (IAS) and its relationship with others variables. MATERIALS AND METHODS: The sample was of 450 patients (383 women, 67 men), age 39.3 + 11.5 years and 31.2 + 5.3 of IMC. IAS of" before" and "during" treatment was calculated with a 24 h-recall previous to the treatment and a Seven-days-dietary-record questionnaire during treatment. The IAS consists of 10 variables that include cereal consumption, fruits, vegetables, dairy products and meats and other variables related to the nutritional guidelines for the Spanish population (SENC, 2004).RESULTS: Habitual dietary habits of the patients were acceptable with an IAS of 67 9 ± 13. However, lipids (43.9 ± 8.4%) and saturated fats (67.4 ± 20.1%) intakes were higher than recommended, while monounsaturated fats were lower (27.8 ± 15.1%). The IAS varied with the BMI and was significantly lower among obese subjects (65.1 ± 11.6) as compared to overweight (69.2 ± 13.9) (P < 0.05). Diet during the treatment, significantly improved with an IAS of 91.4 ± 9.8). IAS of the women studied was better (92.3 ± 9.0) than the one of men (86.4 ± 11.8) (P < 0.05). Patients who reached the goal in weight loss acquired better values of IAS (92.1 ± 9.2) during the treatment that those that did not reach it (87.9 ± 11.7) (P < 0.05).CONCLUSION: In this Spanish population, the diet studied, is useful to promote weight loss through the introduction of changes in dietary habits towards the reincorporation of the Mediterranean cultural tradition (AU)

[Quality of the diet "before and during" a weight loss treatment based on Mediterranean Diet; behavioural therapy and nutritional education]. / Calidad de la dieta "antes y durante" un tratamiento de pérdida de peso basado en una dieta mediterránea; terapia conductual y educación nutricional.

INTRODUCTION: The "Garaulet" Method (Mediterranean Diet, therapy of behavior and nutritional education), has shown to be effective in the treatment of the obesity. OBJECTIVES: To evaluate and to compare the quality of the diets ingested before and during the treatment by means of Index-of-Feeding-Healthful (IAS) and its relationship with others variables. MATERIALS AND METHODS: The sample was of 450 patients (383 women, 67 men), age 39.3 + 11.5 years and 31.2 + 5.3 of IMC. IAS of" before" and "during" treatment was calculated with a 24 h-recall previous to the treatment and a Seven-days-dietary-record questionnaire during treatment. The IAS consists of 10 variables that include cereal consumption, fruits, vegetables, dairy products and meats and other variables related to the nutritional guidelines for the Spanish population (SENC, 2004). RESULTS: Habitual dietary habits of the patients were acceptable with an IAS of 67 9 ± 13. However, lipids (43.9 ± 8.4%) and saturated fats (67.4 ± 20.1%) intakes were higher than recommended, while monounsaturated fats were lower (27.8 ± 15.1%). The IAS varied with the BMI and was significantly lower among obese subjects (65.1 ± 11.6) as compared to overweight (69.2 ± 13.9) (P < 0.05). Diet during the treatment, significantly improved with an IAS of 91.4 ± 9.8). IAS of the women studied was better (92.3 ± 9.0) than the one of men (86.4 ± 11.8) (P < 0.05). Patients who reached the goal in weight loss acquired better values of IAS (92.1 ± 9.2) during the treatment that those that did not reach it (87.9 ± 11.7) (P < 0.05). CONCLUSION: In this Spanish population, the diet studied, is useful to promote weight loss through the introduction of changes in dietary habits towards the reincorporation of the Mediterranean cultural tradition.

Introducción: El método de pérdida de peso (dieta mediterránea, terapia de comportamiento y educación nutricional) ha mostrado ser efectivo en el tratamiento de la obesidad. Objetivo: El objetivo del presente trabajo es evaluar y comparar la calidad de las dietas ingeridas antes y durante el tratamiento mediante el Índice de Alimentación Saludable (IAS) y su relación con diferentes variables. Materiales y métodos: La muestra fue de 392 pacientes (330 mujeres, 62 hombres), edad 39,3 ± 11,5 años y IMC de 31,2 ± 5,3 kg/m2. A partir del recuerdo-24 h previo al tratamiento y del registro dietético 7 días se estimó el IAS de "antes" y "durante" tratamiento. El IAS consta de 10 variables que representan el cumplimiento de objetivos nutricionales para la población española (SENC, 2004). Resultados: Dieta previa, presentó un IAS "necesita mejorar" (68,6 ± 11,6) con lípidos (%) (43,9 ± 8,4) y AGS (% lípidos) (67,4 ± 20,1) elevados, además el contenido en AGM (% lípidos) (27,8 ± 15,1) fue insuficiente. El IAS varió en función del IMC siendo el de obesos inferior al de personas con sobrepeso (65,1 ± 11,6 vs 69, 2 ± 13,9; P < 0,05). La dieta ingerida durante el tratamiento mejoró notablemente IAS (91,4 ± 9,7). El IAS de las mujeres fue superior (92,3 ± 9,1) al de los hombres (84,4 ± 12,0) (P < 0,05). Aquellos que alcanzaron la meta de pérdida de peso adquirieron mejores valores de IAS durante el tratamiento que los que no la alcanzaron (92,1 ± 9,2 vs 87,9 ± 11,7) (P < 0,05). Conclusiones: Según el IAS, la calidad de la dieta estudiada durante el tratamiento de pérdida de peso mejoró significativamente en relación a la dieta habitual del paciente. El IAS de la dieta durante el tratamiento se asocia con el sexo, el estado ponderal (sobrepeso y obesidad) y con el éxito del tratamiento (> 5% de pérdida del peso inicial).

Tissue-specific interaction of Per1/2 and Dec2 in the regulation of fibroblast circadian rhythms.

In mammals, the molecular circadian clockwork is comprised of interlocked transcriptional-translational feedback loops (TTLs). Three Period (Per1-3) and 2 Dec (Dec1/2) genes interact in regulating the activity of the transcriptional activators CLOCK/NPAS2 and BMAL1. While deletion of Per1 and Per2 in mice results in behavioral arrhythmicity, Dec deletion has less dramatic effects on activity rhythms, affecting primarily phase of entrainment and free-running period. In intact animals, clock gene mutant phenotypes are often masked due to intercellular coupling mechanisms that stabilize cellular rhythms. Therefore, to study Per/Dec genetic interaction at the cellular level, we isolated fibroblasts from different tissues of Per1, Per2, and Dec2 single and double mutant mice. We show that in the cellular TTL, Pers and Dec2 act in a principally synergistic way, but tissue-specific differences in this interaction are seen. A rescue of rhythmicity in Per2 mutant cells after additional deletion of Dec2 was observed, indicating that in the absence of Per2, DEC2 destabilizes TTL function. Rhythm power in Per1/Dec2 and Per2/Dec2 double mutants was strongly reduced, suggesting that interaction of Dec2 with both Per genes is important for stabilizing clock period. Contrary to what was observed for behavior, nonsynergistic effects of Dec2 and Per1/2 mutations were observed on cellular clock phase regulation that do not correlate with period effects. Our data reveal cell type-specific interactions of Per1/2 and Dec2 in the regulation of period, phase, and rhythm sustainment, emphasizing the differential organization of the mammalian clock machinery in different tissues.

Ghrelin, sleep reduction and evening preference: relationships to CLOCK 3111 T/C SNP and weight loss.

BACKGROUND: Circadian Locomotor Output Cycles Kaput (CLOCK), an essential element of the positive regulatory arm in the human biological clock, is involved in metabolic regulation. The aim was to investigate the behavioral (sleep duration, eating patterns and chronobiological characteristics) and hormonal (plasma ghrelin and leptin concentrations) factors which could explain the previously reported association between the CLOCK 3111T/C SNP and weight loss. METHODOLOGY/PRINCIPAL FINDINGS: We recruited 1495 overweight/obese subjects (BMI: 25-40 kg/m(2)) of 20-65 y. who attended outpatient obesity clinics in Murcia, in southeastern Spain. We detected an association between the CLOCK 3111T/C SNP and weight loss, which was particularly evident after 12-14 weeks of treatment (P = 0.038). Specifically, carriers of the minor C allele were more resistant to weight loss than TT individuals (Mean±SEM) (8.71±0.59 kg vs 10.4±0.57 kg) C and TT respectively. In addition, our data show that minor C allele carriers had: 1. shorter sleep duration Mean ± SEM (7.0±0.05 vs 7.3±0.05) C and TT respectively (P = 0.039), 2. higher plasma ghrelin concentrations Mean ± SEM (pg/ml) (1108±49 vs 976±47)(P = 0.034); 3. delayed breakfast time; 4. evening preference and 5. less compliance with a Mediterranean Diet pattern, as compared with TT homozygotes. CONCLUSIONS/SIGNIFICANCE: Sleep reduction, changes in ghrelin values, alterations of eating behaviors and evening preference that characterized CLOCK 3111C carriers could be affecting weight loss. Our results support the hypothesis that the influence of the CLOCK gene may extend to a broad range of variables linked with human behaviors.

APOA5 gene variation interacts with dietary fat intake to modulate obesity and circulating triglycerides in a Mediterranean population.

APOA5 is one of the strongest regulators of plasma TG concentrations; nevertheless, its mechanisms of action are poorly characterized. Genetic variability at the APOA5 locus has also been associated with increased cardiovascular disease risk; however, this predisposition could be attenuated in the context of a prudent diet as traditionally consumed in the Mediterranean countries. We have investigated the interaction between a single nucleotide polymorphism (SNP) at the APOA5 gene (-1131T > C) and dietary fat that may modulate TG-rich lipoprotein concentrations and anthropometric measures in overweight and obese participants. We recruited 1465 participants from a Spanish population (20-65 y old; BMI 25-40 kg/m(2)) attending outpatient obesity clinics. Consistent with previous reports, we found an association between the APOA5-1131T > C SNP and TG-rich lipoprotein concentrations that were higher in carriers of the minor allele than in noncarriers (P < 0.001). Moreover, we found a significant genotype-dietary fat interaction for obesity traits. Participants homozygous for the -1131T major allele had a positive association between fat intake and obesity, whereas in those carrying the APOA5-1131C minor allele, higher fat intakes were not associated with higher BMI. Likewise, we found genotype-dietary fat interactions for TG-rich lipoproteins (P < 0.001). In conclusion, we have replicated previous gene-diet interactions between APOA5 -1131T > C SNP and fat intake for obesity traits and detected a novel interaction for TG-rich lipoprotein concentrations. Our data support the hypothesis that the minor C-allele may protect those consuming a high-fat diet from obesity and elevated concentrations of TG-rich lipoproteins.