The Molecular Switching Mechanism at the Conserved D(E)RY Motif in Class-A GPCRs.

The disruption of ionic and H-bond interactions between the cytosolic ends of transmembrane helices TM3 and TM6 of class-A (rhodopsin-like) G protein-coupled receptors (GPCRs) is a hallmark for their activation by chemical or physical stimuli. In the bovine photoreceptor rhodopsin, this is accompanied by proton uptake at Glu(134) in the class-conserved D(E)RY motif. Studies on TM3 model peptides proposed a crucial role of the lipid bilayer in linking protonation to stabilization of an active state-like conformation. However, the molecular details of this linkage could not be resolved and have been addressed in this study by molecular dynamics (MD) simulations on TM3 model peptides in a bilayer of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC). We show that protonation of the conserved glutamic acid alters the peptide insertion depth in the membrane, its side-chain rotamer preferences, and stabilizes the C-terminal helical structure. These factors contribute to the rise of the side-chain pKa (> 6) and to reduced polarity around the TM3 C terminus as confirmed by fluorescence spectroscopy. Helix stabilization requires the protonated carboxyl group; unexpectedly, this stabilization could not be evoked with an amide in MD simulations. Additionally, time-resolved Fourier transform infrared (FTIR) spectroscopy of TM3 model peptides revealed a different kinetics for lipid ester carbonyl hydration, suggesting that the carboxyl is linked to more extended H-bond clusters than an amide. Remarkably, this was seen as well in DOPC-reconstituted Glu(134)- and Gln(134)-containing bovine opsin mutants and demonstrates that the D(E)RY motif is a hydrated microdomain. The function of the D(E)RY motif as a proton switch is suggested to be based on the reorganization of the H-bond network at the membrane interface.

Fijación de carbono atmosférico en la biomasa total de sistemas de producción de cacao en el departamento del tolima, colombia

Los sistemas de uso del suelo con leñosas perennes, como cacaotales, mitigan el cambio climático, al capturar CO2 atmosférico. El objetivo del estudio fue estimar la fijación de carbono en biomasa total en los sistemas de producción de cacao, dominantes del Tolima, Colombia. El estudio, se realizó en Rovira y Falan, empleando un diseño experimental, completamente al azar, con seis sistemas de producción de cacao: monocultivo, sistema agroforestal (SAF) con maderables, SAF con aguacate, SAF con cítricos, SAF con frutales, y SAF con maderables y frutales y tres repeticiones. Se establecieron dos parcelas de muestreo rectangulares, de 1.000m² por unidad de muestreo, donde se identificaron y midieron los árboles con dap ≥ 10cm (diámetro del tronco a la altura del pecho -dap- y altura total -ht). Se establecieron dos sub-parcelas de 256m², por parcela principal, para medir arbustos de cacao: ht y diámetro del tronco, a 30 cm de altura (D30). Se emplearon modelos alométricos, para estimar la biomasa. El SAF con maderables y frutales y SAF con frutales presentaron la mayor biomasa total (122,0 y 72,5t/ ha). En el SAF con maderables y frutales, se evidenció el mayor almacenamiento de carbono, con 61,0t C/ha, mientras que la mayor tasa de fijación de carbono fue de 17,7t/ha/ año, para el SAF con cítricos. Los resultados sugieren que la producción de cacao, que incluyen otras especies vegetales, diversifica la producción e incrementa la seguridad alimentaria y la generación de servicios ambientales, tal como la captura de carbono atmosférico.

The land use systems with woody perennials species, such as cacao plantations, mitigate climate change by capturing atmospheric CO2. The aim of the study was to estimate the fixation of atmospheric carbon in total biomass in the most dominant cocoa production systems in Tolima, Colombia. The study was carried out in Rovira and Falan using a completely randomized experimental design with six cocoa production systems: monoculture, agroforestry system (SAF) with timber trees, SAF with avocado, SAF with citrus species, SAF with fruit trees and SAF with timber and fruit species with three replications. Two rectangular sampling plots 1000m² were established by sampling unit where trees with diameter of trunk at breast height (dbh) ≥ 10cm were identified and measured (dbh and total height - th). Two subplots of 256 m² by main plot, for measuring cocoa bushes were established: ht and trunk diameter at 30cm height (D30). Allometric models to estimate biomass were used. The AFS with timber and fruit species and AFS with fruit species had the greatest biomass (122.0 and 72.5t/ha). In the AFS with timber and fruit species presented the highest carbon storage in biomass with 61.0t C/ha; whereas the greatest carbon fixation rate was 17.7 t/ha/year in AFS with citrus species. The results suggest that cocoa plantations that include other plant species diversify the production and increase food security and generation of environmental services, such as capturing atmospheric carbon.

Conjugated estrogens and tibolone modify the gonadotrophin glycosylation pattern in postmenopausal women.

OBJECTIVE: The purpose of this study was to evaluate the changes in the percentage of glycosylation of FSH and LH when using conjugated estrogens and tibolone as hormonal therapy (HT) in postmenopausal compared with regular menstrual cycles. DESIGN: The study had three groups, with 10 participants in each group. The control group consisted of 10 women with normal menstrual cycles, a second group with 10 postmenopausal patients who received conjugated estrogens (Premarin 0.625 mg), and a third group had 10 patients who received tibolone (Livial 2.5 mg). All patients had hormonal determinations before and 6 months after the use of HT. Concavalin-A was used to separate the different glycosylated species of gonadotrophins, defined as unbound (UB: triantennary and bisecting oligosaccharides), weakly bound (WB: biantennary and truncated oligosaccharides), and firmly bound isoforms (FB: hybrid- and high-mannose-type oligosaccharides) in patients serum. Elusions containing the fractions with the isoforms were concentrated using Centriprep membranes (Amicon, Beverly, Mass., USA) and the different gonadotrophins isoforms were quantified by chemiluminescence using an automated system Immulite. RESULTS: The pattern of distribution of gonadotrophins glycosylation in patients treated with conjugated estrogens and tibolone was similar to that observed in the control group with normal menstrual cycles, although with different glycosylation percentage. When the glycosylation percentages were compared for FSH, in the fraction UB the glycosylation did not have significant differences, but the fraction WB that has isoforms with high amounts of oligosaccharides showed a different percentage of glycosylation, where the recovery percentage was bigger with tibolone use (p < 0.05). The FB fraction had similar hormonal glycosylation with estrogens and tibolone use. When compared, LH glycosylation showed significant differences (p < 0.05) in the fraction UB with a bigger glycosylation degree, and in the fraction FB with a smaller glycosylation percentage, both in the tibolone group compared with conjugated estrogens use. CONCLUSIONS: Our results show that there are different percentages of glycosylation of gonadotrophins depending on type of HT (conjugated estrogens vs tibolone), and this could be of utility in young women with ovarian problem since to know the isoform that more favors to cellular activity could help a better therapeutic.

Comportamiento reológico de harinas y almidones de yuca (Manihot esculenta Cranz) durante un proceso de extrusión / Rheological modeling of cassava (Manihot esculenta Crantz) starches and flours during an extrusion process

La obtención de modelos reológicos para almidones y harinas de yuca durante un proceso de extrusión se realizó con el fin de predecir los efectos de la velocidad de corte, la temperatura y el contenido de agua en la viscosidad de la masa fundida. La experimentación se llevó a cabo en un reómetro capilar de alta presión (Rosand RH-7) de doble boquilla y los datos de viscosidad obtenidos se ajustaron al modelo reológico de Harper. Todos los materiales presentaron un comportamiento característico de disminución de la viscosidad al aumentar la velocidad de corte. La viscosidad de los almidones fue más sensible a las modificaciones en las condiciones de proceso, como el cambio de temperatura y contenido de agua; por otra parte, la presencia de otros componentes, como fibra y proteína tuvo una alta influencia en la viscosidad de las harinas, lo que se explica por la interferencia de estos compuestos en el proceso de gelatinización del almidón.