Biomedical subjects

Carlos Conde

Publications and source records attributed to Carlos Conde.

4 recordsLinked to original sources

PubMed2006

Utilization and transport of mannitol in Olea europaea and implications for salt stress tolerance.

Mannitol is one of the primary photosynthetic products and the major phloem-translocated carbohydrate in Olea europaea L., an important crop in the Mediterranean basin. Uptake of mannitol in heterotrophic cell suspensions of O. europaea was shown to be mediated by a 1 : 1 polyol : H+ symport system with a Km of 1.3 mM mannitol and a Vmax of 1.3 nmol min(-1) mg(-1) DW. Dulcitol, sorbitol and xylitol competed for mannitol uptake, whereas glucose and sucrose did not. Reverse transcription-PCR (RT-PCR) performed on mRNA extracted from cultured cells exhibiting high mannitol transport activity allowed the cloning of a partial O. europaea mannitol carrier OeMaT1. The Vmax of mannitol uptake and the amount of OeMaT1 transcripts increased along with polyol depletion from the medium, suggesting that the mannitol transport system may be regulated by its own substrate. Addition of 100-500 mM NaCl to cultured cells enhanced the capacity of the polyol : H+ symport system and the amount of OeMaT1 transcripts, whereas it strongly repressed mannitol dehydrogenase activity. Measurements of cell viability showed that mannitol-grown cells remained viable 24 h after a 250 and 500 mM NaCl pulse, whereas extensive loss of cell viability was observed in sucrose-grown cells. OeMaT1 transcripts increased throughout maturation of olive fruits, suggesting that an OeMaT is involved in the accumulation of mannitol during ripening of olive. Thus, mannitol transport and compartmentation by OeMaT are important to allocate this source of carbon and energy, as well as for salt tolerance and olive ripening.

Biological Transport↗

PubMed2006

Pathways of glucose regulation of monosaccharide transport in grape cells.

Grape (Vitis vinifera) heterotrophic suspension-cultured cells were used as a model system to study glucose (Glc) transport and its regulation. Cells transported D-[14C]Glc according to simple Michaelis-Menten kinetics superimposed on first-order kinetics. The saturating component is a high-affinity, broad-specificity H+ -dependent transport system (Km = 0.05 mm). Glc concentration in the medium tightly regulated the transcription of VvHT1 (Vitis vinifera hexose transporter 1), a monosaccharide transporter previously characterized in grape berry, as well as VvHT1 protein amount and monosaccharide transport activity. All the remaining putative monosaccharide transporters identified so far in grape were poorly expressed and responded weakly to Glc. VvHT1 transcription was strongly repressed by Glc and 2-deoxy-D-Glc, but not by 3-O-methyl-D-Glc or Glc plus mannoheptulose, indicating the involvement of a hexokinase-dependent repression. 3-O-Methyl-D-Glc, which cannot be phosphorylated, and Glc plus mannoheptulose induced a decrease of transport activity caused by the reduction of VvHT1 protein in the plasma membrane without affecting VvHT1 transcript levels. This demonstrates hexokinase-independent posttranscriptional regulation. High Glc down-regulated VvHT1 transcription and Glc uptake, whereas low Glc increased those parameters. Present data provide an example showing control of plant sugar transporters by their own substrate both at transcriptional and posttranscriptional levels. VvHT1 protein has an important role in the massive import of monosaccharides into mesocarp cells of young grape berries because it was localized in plasma membranes of the early developing fruit. Protein amount decreased abruptly throughout fruit development as sugar content increases, consistent with the regulating role of Glc on VvHT1 expression found in suspension-cultured cells.

Biological Transport↗

PubMed2006

The non-host pathogen Botrytis cinerea enhances glucose transport in Pinus pinaster suspension-cultured cells.

Botrytis cinerea is the causal agent of grey mould disease and a non-host necrotrophic pathogen of maritime pine (Pinus pinaster). Recent evidence suggests that pathogen challenge can alter carbon uptake in plant cells; however, little is known on how elicitor-derived signalling pathways control sugar transport activity. P. pinaster suspended cells are able to absorb D-[14C]glucose with high affinity, have an H+-dependent transport system (Km, 0.07 mM; Vmax, 1.5 nmol min(-1) mg(-1) DW), are specific for D-glucose, D-fructose, D-galactose and D-xylose, and are subject to glucose repression. When elicited by B. cinera spores, suspended cells exhibit calcium-dependent biphasic reactive oxygen species (ROS) production, the second burst also being dependent on NADPH oxidase, mitogen-activated protein kinase (MAPK), and de novo transcription and protein synthesis. Challenging suspended cells incubated in sugar-free medium resulted in an up to 3-fold increase in glucose transport capacity over non-elicited cultures 24 h after elicitation, and a 14-fold increase over elicited cells incubated with 2% glucose. Enhanced glucose uptake depended on NADPH oxidase and calcium influx, but not MAPK. In contrast, the increase of glucose transport activity induced by sugar starvation was dependent on the activation of MAPK but not NADPH oxidase. Both responses appeared to be dependent on de novo transcription and protein synthesis.

Biological Transport↗

PubMed2002

Blend uniformity analysis using stream sampling and near infrared spectroscopy.

A near infrared spectroscopic method was developed to determine drug content in a 20% (wt/wt) ibuprofen and spray-dried hydrous lactose blend. A blending profile was obtained after blending for 0.5, 1, 3, 5, 10, and 20 minutes. Stream sampling was used to collect about 20 blend samples at each of the blending times from a laboratory scale V-blender. The samples collected were used to develop a near infrared calibration model. The calibration model was then used to determine the drug content of unknown samples from 2 validation blends. The validation blends were not included in the calibration model; they were used to evaluate the effectiveness of the calibration model. A total of 45 samples from the 2 validation blends were predicted by the near infrared calibration model and then analyzed by a validated UV spectrophotometric method. The root mean square error of prediction for the first validation blend was 5.69 mg/g and 3.30 mg/g for the samples from the second blend. A paired t test at the 95% confidence level did not indicate any differences between the drug content predicted by the near infrared spectroscopy (NIRS) method and the validated UV method for the 2 blends. The results show that the NIRS method could be developed while the blending profile is generated and used to thoroughly characterize a new formulation during development by analyzing a large number of samples. The new formulation could be transferred to a manufacturing plant with an NIRS method to facilitate blend uniformity analysis.

Calibration↗